See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/242178938

Estimating Soil Loss Using Universal Soil Loss Equation (USLE) for Soil
Conservation planning at Medego Watershed, Northern Ethiopia

Article in Journal of American Science · January 2009

CITATIONS READS

112 5,222

2 authors:

Gebryesus Brhane Tesfahunegn Kirubel Mekonnen

Aksum University 18 PUBLICATIONS 545 CITATIONS
60 PUBLICATIONS 1,603 CITATIONS
SEE PROFILE
SEE PROFILE

All content following this page was uploaded by Kirubel Mekonnen on 08 October 2020.

The user has requested enhancement of the downloaded file.

The Journal of American
Science

,661

Volume 5 - Number 1 (Cumulated No. 16), January 1, 2009

Marsland Press, Michigan, The United States

http://www.americanscience.org editor@americanscience.org
 Journal of American Science ISSN 1545-1003

Journal of American Science

The Journal of American Science is an international journal with a purpose to enhance our natural and scientific knowledge
dissemination in the world under the free publication principle. Any valuable paper that describes natural phenomena and existence or
any report that conveys scientific research and pursuit are welcome, including both natural and social sciences. Papers submitted could
be reviews, objective descriptions, research reports, opinions/debates, news, letters, and other types of writings that are nature and
science related. The journal is calling for papers and seeking co-operators and editors as well.
Editor-in-Chief: Hongbao Ma
Associate Editors-in-Chief: Shen Cherng, Jingjing Z Edmondson, Qiang Fu, Yongsheng Ma
Editors: George Chen, Mark Hansen, Mary Herbert, Deng-Nan Horng, Wayne Jiang, Mark Lindley, Margaret D. Ma, Mike Ma, Da
Ouyang, Xiaofeng Ren, Ajaya Kumar Sahoo, Shufang Shi, Tracy X Qiao, Pankaj Sah, George Warren, Qing Xia, Yonggang Xie, Lijian
Yang, Jenny Young, Tina Zhang, Ruanbao Zhou, Yi Zhu
Web Design: Jenny Young

Introductions to Authors
1. General Information Reference Examples:
(1) Goals: As an international journal published both in print and on Journal Article: Hacker J, Hentschel U, Dobrindt U. Prokaryotic
internet, the Journal of American Science is dedicated to the chromosomes and disease. Science 2003;301(34):790-3.
dissemination of fundamental knowledge in all areas of nature and Book: Berkowitz BA, Katzung BG. Basic and clinical evaluation of
science. The main purpose of the Journal of American Science is to new drugs. In: Katzung BG, ed. Basic and clinical pharmacology.
enhance our knowledge spreading in the world under the free Appleton & Lance Publisher. Norwalk, Connecticut, USA. 1995:60-9.
publication principle. It publishes full-length papers (original (10) Submission Address: editor@sciencepub.net, Marsland Press,
contributions), reviews, rapid communications, and any debates and P.O. Box 21126, Lansing, Michigan 48909, The United States, 517-
opinions in all the fields of nature and science. 349-2362.
(2) What to Do: The Journal of American Science provides a place (11) Reviewers: Authors are encouraged to suggest 2-8 competent
for discussion of scientific news, research, theory, philosophy, reviewers with their name and email.
profession and technology - that will drive scientific progress.
Research reports and regular manuscripts that contain new and 2. Manuscript Preparation
significant information of general interest are welcome. Each manuscript is suggested to include the following components but
(3) Who: All people are welcome to submit manuscripts in any fields authors can do their own ways:
of nature and science. (1) Title page: including the complete article title; each author’s full
(4) Distributions: Web version of the journal is freely opened to the name; institution(s) with which each author is affiliated, with city,
world, without any payment or registration. The journal will be state/province, zip code, and country; and the name, complete mailing
distributed to the selected libraries and institutions for free. For the address, telephone number, facsimile number (if available), and e-mail
subscription of other readers please contact with: address for all correspondence.
editor@americanscience.org or americansciencej@gmail.com or (2) Abstract: including Background, Materials and Methods, Results,
editor@sciencepub.net and Discussions.
. (3) Key Words.
(5) Advertisements: The price will be calculated as US$400/page, i.e. (4) Introduction.
US$200/a half page, US$100/a quarter page, etc. Any size of the (5) Materials and Methods.
advertisement is welcome. (6) Results.
(7) Discussions.
2. Manuscripts Submission (8) Acknowledgments.
(1) Submission Methods: Electronic submission through email is (9) Correspondence to.
encouraged and hard copies plus an IBM formatted computer diskette (10) Submission date
would also be accepted. (11) References.
(2) Software: The Microsoft Word file will be preferred.
(3) Font: Normal, Times New Roman, 10 pt, single space. Journal Address:
(4) Indent: Type 4 spaces in the beginning of each new paragraph. Marsland Press
(5) Manuscript: Don’t use “Footnote” or “Header and Footer”. 2158 Butternut Drive
(6) Cover Page: Put detail information of authors and a short title in Okemos, MI 48864
the cover page. The United States
(7) Title: Use Title Case in the title and subtitles, e.g. “Debt and Telephone: (517) 349-2362
Agency Costs”. E-mail: editor@americanscience.org;
(8) Figures and Tables: Use full word of figure and table, e.g. editor@sciencepub.net;
“Figure 1. Annul Income of Different Groups”, Table 1. Annual americansciencej@gmail.com
Increase of Investment”. Websites: http://www.americanscience.org;
(9) References: Cite references by “last name, year”, e.g. “(Smith, http://www.sciencepub.org
2003)”. References should include all the authors’ last names and
initials, title, journal, year, volume, issue, and pages etc.

¤ 2009 Marsland Press

http://www.americanscience.org; http://www.sciencepub.org i editor@sciencepub.net

 The Journal of American Science
Volume 5 - Number 1, January 10, 2009, ISSN 1545-1003
Contents
1. Plant Diversity of a Fresh Water Swamp of Doon Valley, India.
Manhas R.K., Mukesh Kumar Gautam and Deepa Kumari 1-7
2. Histological Interactions Of Paecilomyces Lilacinus And Meloidogyne Incognita
On Bitter Gourd Mohd Yaqub Bhat, Hissamuddin, Nazir Ahmad Bhat 8-12
3. Experimental Study on The Dynamic Behaviors of the Material for Clay core wall sand dams
Xu Shangjie,Dang Faning,Tian Wei,Cheng Mo 13-18

4. Influence of Subchronic Exposure of Profenofos on Biochemical

Markers and Microelements in Testicular Tissue of Rats
Afaf A. El-Kashoury 19-28
5. Effect of Duckweed meal on the rate of mold infestation in stored pelleted fish feed.
B.N. Effiong, A. Sanni 29-34
6. Distribution and Sources of Organochlorine Pesticides (OCPs)
in Karst Cave, Guilin, China
Annette Sylvie Muhayimana, Qi Shihua, Wang Yinghui 35-43
7. An Incidence of Substratum Discolouration in a Tropical West African Lagoon.
Onyema, I.C. and Nwankwo, D.I. 44-48
8. The Influences of Extremely Low Frequency AC Magnetic Fields
At 60Hz on Mung Beans Growth
Pai-Tsun Tien and Show-Ran Wang 49-54
9. The Inflation Dynamics of the ASEAN-4: A Case Study of the Phillips Curve Relationship
Klarizze Anne M. Puzon 55-57
10. Estimating Soil Loss Using Universal Soil Loss Equation (USLE)
for Soil Conservation planning at Medego Watershed, Northern Ethiopia
Gebreyesus Brhane and Kirubel Mekonen 58-69
11. Conservation through in vitro method: A case of plant regeneration
through somatic embryogenesis in Quercus semecarpifolia Sm.
Sushma Tamta, Lok Man S. Palni, P. Vyas and M.S.Bisht 70-76
12. Synthesis, characterization and Electroluminescence of
BPh2(2-(benzimidazol-2-yl) pyridinato) compound
Anchi Yeh 77-82
13. White Organic Electroluminescence Base on a new Aluminum Complex
Anchi Yeh, Hsien-Chiao Teng 83-87

© 2009 Marsland Press, the United States, editor@sciencepub.net

Marsland Press
Journal of American Science 2009: 5(1), 1-7
_____________________________________________________________________________________________
Research Article
Plant Diversity of a Fresh Water Swamp of Doon Valley, India.
1
Manhas, R.K., 2Mukesh Kumar Gautam and 3Deepa Kumari
1
Department of Botany, S.P. College, Srinagar- 190 001 (J and K)
2
Forest Ecology and Environment Division, Forest Research Institute, D.Dun- 248 006 (UA)
3
Cosmic Hearts Higher Secondary School, Kathua- 184 101 (J and K)
________________________________________________________________________________________
ABSTRACT: The present study was conducted in a highly degraded and fragmented swamp of Doon valley,
India. A total of 162 plant species were recorded from the swamp. Dicotyledons contributed 71%,
monocotyledons 23.5% and pteridophytes 5.6%. Poaceae with 15 genera and 17 species was the most
represented family. Biological spectrum of the present study site shows that therophytes were the most
dominating life-form of the swamp, representing high anthropogenic disturbance in the region and limited
niche space for the vegetation. [Journal of American Science 2009: 5(1), 1-7] (ISSN: 1545-1003)

Key words: Dicotyledons, swamp

1. INTRODUCTION latitude and 78º 05ƍ 55ƍƍ E longitude. Most of the

Fresh water swamps are the unique ecosystems having swamp is urbanized. A very few patches of swampy
very specific vegetation. These are sites of natural vegetation are present here and there along the river
succession and therefore contain all the groups of plant Dholani, a tributary of the Song river.
kingdom in a single place. Water is the prime requisite
of the vegetation of the swamp forests and any Methodology
alteration in the availability of water affects their Plant specimens were collected, dried, poisoned and
presence as well as distribution. Doon valley, situated mounted on the herbarium sheets. Standard methods
at the foothills of the Himalaya between rivers the given in Jain and Rao (1977) for collection,
Yamuna and the Ganges, use to have a chain of preservation and maintenance of specimen in
swamps (Manhas et al., 2007). But due to herbarium were followed. Herbariums of Forest
anthropogenic activities these forests are disappearing Research Institute and Botanical Survey of India,
at a very fast rate. Nakraunda is one of the most Northern Circle were consulted for the identification of
degraded swamps of Doon valley. Most of the area of each species. Floras written by Babu (1980) and
the swamp has been converted to agriculture fields and Kanjilal (1901) were used for the nomenclature of the
residential colonies. species. These plant species were further classified;
first on the basis of habit and then on basis of life-
Taxonomic study of swamp forests of Doon forms as defined by Raunkiaer (1934).
valley was first carried out by Kanjilal in 1901, since
then a number of studies have been conducted by 3. RESULTS
various workers for floristic diversity (Dakshini, 1960a, A total of 162 plant species were found in the present
1960b, 1965, 1970 and 1974; Dhyani and Joshi, 2007; study site (Table 1). The contribution of dicotyledons
Sharma and Joshi, 2008), successional studies (Som was 71.0%, monocotyledons 23.5% and pteridophytes
Deva and Srivastava, 1978; Srivastava et al., 2000) and 5.6%. Table 2 reveals that Poaceae (15 genera/ 17
community dynamics (Manhas et al., 2007; Kandwal et species) was the most dominating family of Nakraunda
al., 2007). In the present paper we have studied floristic swamp. The other important families were Asteraceae
and life-form diversity of Nakraunda swamp forest of (11 genera/ 12 species), Acanthaceae (10 genera/ 11
Doon valley. species), Cyperaceae (6 genera/ 9 species) and
Scrophulariaceae (4 genera/ 7 species). Cyperus and
2. MATERIALS AND METHODS Polygonum, both having three species, were the most
represented genera. Classification on the basis of habit
Study Site (Figure 1) shows that herbs were the main vegetation
Nakraunda swamp is situated about 15 km east of form with 44.4% contribution followed by shrubs
Dehradun on Dehradun-Doiwala road at 30º 14ƍ 15ƍƍ N (15.4%) and grasses (10.5%).

1
Plant Diversity of a Fresh Water Swamp of Doon Valley, India.
Manhas, R.K. et al.
_____________________________________________________________________________________________________________________

Table 1
Floristic diversity in the Nakraunda swamp and its comparison with other swamps of Doon valley

Plant Groups Families Genera Species References

Angiosperms 53 130 155 Dhyani and Joshi (2007)
Angiosperms 71 218 278 Sharma and Joshi (2008)
Total (1 + 2) 61 141 162 Present study
1. Angiosperms (i + ii) 55 135 153
(i) Dicotyledons 45 103 115
(ii) Monocotyledons 10 32 38
2. Pteridophytes 6 6 9

Undershrubs (1.9%)
Small trees (0.6%)
Ferns (5.6%)
Sedges (5.6%)
Climbers (7.4%)

Trees (8.6%)

Grasses (10.5%)
Herbs (44.4%) Shrubs (15.4%)

Fig 1: Pie diagram showing percentage contribution of various plant habits.

50
Raunkiaer (1934) Present Study
45
Percentage Contribution

40
35
30
25
20
15
10
5
0
Phanerophytes Chamaephytes Hemicryptophytes Cryptophytes T herophytes
Life-Forms
Fig 2: Biological spectrum of life-forms of present study and
its comparison with the Raunkiaer’s normal biological spectrum representing world flora.

2
Marsland Press
Journal of American Science 2009: 5(1), 1-7
_____________________________________________________________________________________________

Biological spectrum of the swamp was also the most characteristic life-form of the present study as
studied (Figure 2) and compared with the Raunkiaer’s compared to phanerophytes in Raunkiaer’s normal
normal biological spectrum (Raunkiaer, 1934) biological spectrum.
representing the world flora. Therophytes (45.1%) were

Table 2
List of plant species present in the Nakraunda swamp along with family, habit and life-form.
The life-forms mentioned in the table are: Ph = Phanerophytes; Ch = Chamaephytes; He = Hemicryptophytes;
Cr = Cryptophytes; and Th = Therophytes (for definitions see Raunkiaer, 1934).
Plant Species Family Habit Life-form
Achyranthes aspera Linn. Acanthaceae Herb Th
Acorus calamus Linn. Araceae Herb Cr
Adenostemma lavenia (Linn.) O. Kuntze. Asteraceae Shrub Ch
Adhatoda vasica Nees Acanthaceae Shrub Ch
Adiantum capillus-veneris Linn. Adiantaceae Fern Cr
Adiantum incisum Forssk. Adiantaceae Fern Cr
Aerva sanguinoleata (Linn.) DC. Amaranthaceae Herb Th
Aerva scandens Wall. Amaranthaceae Herb Th
Ageratum conyzoides Linn. Asteraceae Herb Th
Alternanthera sessilis R. Br. Amaranthaceae Herb Th
Amaranthus spinosus Linn. Amaranthaceae Herb Th
Anagallis arvensis Linn. Primulaceae Herb Th
Anisomelas indica Kuntze. Lamiaceae Herb Th
Apium leptophyllum (Pers.) F. Muell. Ex Benth. Apiaceae Herb Th
Arachne cordifolia (Decne) Hurusawa Euphorbiaceae Herb Th
Argemone mexicana Linn. Papavaraceae Herb Th
Arundo donax Linn. Poaceae Grass He
Asclepias curassavica Linn. Asclepiadaceae Shrub Ph
Asparagus racemosus Willd. Liliaceae Herb Ph
Bacopa monniera (Linn.) Wettst. Scrophulariaceae Herb Th
Bacopa procumbens (Mill.) Greenm. Scrophulariaceae Herb Th
Bauhinia vahlii Wight and Arn. Caesalpiniaceae Climber Ph
Bauhinia variegata Linn. Caesalpiniaceae Tree Ph
Belamcanda chinensis (Linn.) DC. Iridaceae Herb Cr
Bidens tripartite Linn. Asteraceae Herb Th
Bischofia javanica Blume Euphorbiaceae Tree Ph
Boehmeria platyphylla D. Don Urticaceae Herb Th
Boerhavia diffusa Linn. Nyctaginaceae Herb Th
Bombax ceiba Linn. Bombaceae Tree Ph
Butea monosperma (Lamk.) Taub. Fabaceae Tree Ph
Caesalpinia decapetala (Roxb.) Alston Caesalpiniaceae Climber Ph
Calamus tenuis Roxb. Palmaceae Shrub Ph
Capparis zeylanica Linn. Capparidaceae Climber Ph
Capsella bursa-pastoris (Linn.) Medic Brassicaceae Herb Th
Carissa opaca stapf. Apocynaceae Shrub Ph
Cassia tora Linn. Caesalpiniaceae Herb Th
Cheilanthes farinosa Blanford Sinopteridaceae Fern Cr
Chenopodium album Linn. Chenopodiaceae Herb Th
Chenopodium ambrosioides Linn. Chenopodiaceae Herb Th
Chloris dolichostachya Lagasca Poaceae Grass He
Clerodendron viscosum Vent. Verbenaceae Undershrub Ph
Coccinea grandis (Linn.) Voigt. Cucurbitaceae Climber Ph
Coix lachrymal-jobi Linn. Poaceae Grass He

3
Plant Diversity of a Fresh Water Swamp of Doon Valley, India.
Manhas, R.K. et al.
_____________________________________________________________________________________________________________________

Commelina benghalensis Linn. Commelinaceae Herb Th

Corchorus acutangular Lamk. Tiliaceae Herb Th
Coronopus didymus (Linn.) J.E. Smith Brassicaceae Herb Th
Crotalaria albida Heyne. Fabaceae Herb Th
Curculigo orchioides Gaertn. Hypoxidaceae Herb Cr
Cyathocline purpurea (D.Don) O. Kuntze Acanthaceae Herb Th
Cynodon dactylon (Linn.) P. Beauv. Poaceae Grass He
Cyperus iriaLinn. Cyperaceae Sedge He
Cyperus niveus Retz. Cyperaceae Sedge He
Cyperus rotundus Linn. Cyperaceae Sedge He
Desmodium trifolium DC. Fabaceae Herb Th
Dicliptera roxburghiana Nees Acanthaceae Herb Th
Digitaria adscendens (HBK) Henr. Poaceae Grass He
Digitaria ischaemum Poaceae Grass He
Dioscorea bulbifera Linn. Dioscoreaceae Climber Ph
Dryopteris cochleata (D.Don) C. Chr. Aspidaceae Fern Cr
Duchesnea indica Focke. Rosaceae Herb Th
Echinochloa colonum (Linn.) Link. Poaceae Grass He
Eclipta prostrata Linn. Asteraceae Herb Th
Emilia sonchifolia DC. Asteraceae Herb Th
Eranthemum nervosum (Vahl) R.Br. Acanthaceae Herb Th
Eriophorum comosum Wall. Cyperaceae Sedge He
Eupatorium adenophorum Spreng. Asteraceae Herb Th
Euphorbia hirta Linn. Euphorbiaceae Herb Th
Ficus palmata Forssk. Moraceae Tree Ph
Ficus riligiosa Linn. Moraceae Tree Ph
Fimbristylis dichotoma (Linn.) Vahl. Cyperaceae Sedge He
Fimbristylis falcata (Vhl) Kunth Cyperaceae Sedge He
Flacourtia indica (Burm. f.) Merr. Flacourtiaceae Shrub Ph
Flemingia bracteata Wight Fabaceae Shrub Ph
Floscopa scandens Lour. Commelinaceae Herb Th
Galinsoga parviflora Cav. Asteraceae Herb Th
Hemigraphis rupestris Heyne ex T. Anders. Acanthaceae Herb Th
Holarrhena antidysentrica Wall. Apocynaceae Small tree Ph
Holoptelia integrifolia (Roxb.) Planch. Ulmaceae Tree Ph
Ichnocarpus frutescens (Linn.) R. Br. Apocynaceae Climber Ph
Imperata cylindrical (Linn.) Beauv. Poaceae Grass He
Ipomoea palmata Forsk. Convolvulaceae Climber Ph
Ipomoea quamoclit Linn. Convolvulaceae Climber Th
Isachne globosa (thumb.) O. Kuntze Poaceae Grass He
Jasminum multiflorum (Burm. f.) Andr. Oleaceae Climber Ph
Juncus bufonius Linn. Cyperaceae Sedge He
Justicia gendarussa Linn. Acanthaceae Shrub Ch
Justicia quinqueqngulqris Koenig ex Roxb. Acanthaceae Shrub Ch
Kyllingia triceps Rottb. Cyperaceae Sedge He
Lantana camara Linn. Verbenaceae Shrub Ph
Lepidagathis cuspidate Nees Acanthaceae Shrub Ph
Lindernia ciliate (Col.) Pennell Scrophulariaceae Herb Th
Lindernia crustata (Linn.) F. Muell. Scrophulariaceae Herb Th
Lygodium flexuosum(Linn.) Sw. Schizaeaceae Fern Cr
Mallotus philippensis Muell. Arg. Euphorbiaceae Tree Ph
Malvaviscus penduliflorus DC. Malvaceae Shrub Ph
Martynia annua Linn Martyniaceae Undershrub Th

4
Marsland Press
Journal of American Science 2009: 5(1), 1-7
_____________________________________________________________________________________________
Mazus pumilus (Burm. f.) Steenis Scrophulariaceae Herb Th
Mentha piperita Linn. Lamiaceae Herb Th
Mimosa pudica Linn. Mimosaceae Shrub Ch
Monochoria vaginalis Presl. Pontederiaceae Herb Th
Morus alba Linn Moraceae Tree Ph
Murraya koenigii (Linn.) Spreng. Rutaceae Shrub Ph
Narenga porphyrocoma (Hance ex Trim.) Bor Poaceae Grass He
Nepeta hindostana (Roth.) Haines Lamiaceae Herb Th
Ocimum gratissimum Linn. Lamiaceae Herb Th
Oenanthe stolonifera DC. Apiaceae Herb Th
Ophioglossum reticulatum Linn. Ophioglossaceae Fern Th
Ophioglossum vulgatum Linn. Ophioglossaceae Fern Th
Oplismenus compositus (Linn.) P. Beauv. Poaceae Grass He
Panicum miliaceum Linn. Poaceae Grass He
Parthenium hysterophorus Linn. Asteraceae Shrub Th
Paspalum distichum Linn. Poaceae Grass He
Passiflora incarnate Linn. Passifloraceae Climber Ph
Phlogacanthus thyrsiformis (Hardw.) Mabb. Acanthaceae Shrub Ch
Phoenix acaulis Buch. Palmaceae Tree Ph
Phragmites karka Trin Poaceae Grass He
Phyla nodiflora Linn. Verbenaceae Herb Th
Phyllanthus niruri Linn. Euphorbiaceae Herb Th
Pilea scripta (Buch.- Ham. ex D.Don) Wedd. Urticaceae Undershrub Ph
Plantago major Linn. Plantaginaceae Herb Th
Plumbago zeylanica Linn. Plumbaginaceae Herb Th
Pogostemon plectranthoides Desf. Lamiaceae Shrub Ch
Polygonum barbatum Linn. Polygonaceae Herb Th
Polygonum hydropiper Linn. Polygonaceae Herb Th
Polygonum plebejum R. Br. Polygonaceae Herb Th
Pouzolzia pentandra (Roxb.) Benn. Polygonaceae Herb Th
Pteris quadriaurita Retz. Pteridaceae Fern Cr
Pteris villata Linn. Pteridaceae Fern Cr
Pyrus pashia Buch.- Ham. ex D. Don Rosaceae Tree Ph
Ranunculus sceleratus Linn. Ranunculaceae Herb Th
Rorripa nasturtium-aquaticum (Linn.) Hayek. Brassicaceae Herb Th
Rotula aquatica Lour. Boraginaceae Herb Th
Rouvolfia serpentine (Linn.) Benth.- ex Kurz. Apocynaceae Shrub Ch
Rubus niveus Thunb. Rosaceae Shrub Ph
Rumex nepalensis Spreng. Polygonaceae Herb Th
Rungia pectinata (Linn.) Nees Acanthaceae Herb Th
Scirpus juncoides Roxb. Cyperaceae Sedge He
Setaria glauca Baeuv. Poaceae Grass He
Shorea robusta Gaertn. f. Dipterocarpaceae Tree Ph
Sida cordifolia Linn. Malvaceae Herb Th
Smilax zeylanica Linn. Liliaceae Climber Ph
Solanum hispidum Pers. Solanaceae Shrub Ph
Solanum torvum Swartz. Solanaceae Shrub Ph
Sporobolus diander Beauv. Poaceae Grass He
Sporobolus indicus R. Br. Poaceae Grass He
Stellaria media Linn. Caryophyllaceae Herb Th
Syzygium cumini (Linn.) Skeel Myrtaceae Tree Ph
Tectona grandis Linn. f. Verbenaceae Tree Ph
Trifolium repens Linn. Fabaceae Herb Th

5
Plant Diversity of a Fresh Water Swamp of Doon Valley, India.
Manhas, R.K. et al.
_____________________________________________________________________________________________________________________

Triumfetta rhomboidea Jacq. Tiliaceae Herb Th

Vallaris solanacea O. Kuntze Apocynaceae Climber Ph
Vernonia anthelmintica Willd. Asteraceae Herb Th
Vernonia cineria (Linn.) Lees. Asteraceae Herb Th
Veronica agrestis H.K.f. Scrophulariaceae Herb Th
Veronica anagallis Linn. Scrophulariaceae Herb Th
Vicia sativa Linn. Fabaceae Herb Th
Vitex negundo Linn. Verbenaceae Shrub Ph
Woodfordia fruticosa (Linn.) Kurtz. Lythraceae Shrub Ph
Xanthium strumarium Linn. Asteraceae Shrub Th
Youngia japonica DC. Asteraceae Herb Th
Zeuxine strateumatica (Linn.) Schltr. Orchidaceae Herb Cr
Ziziphus mauritiana Lam. Rhamnaceae Shrub Ch

4. DISCUSSION REFERENCES
The floristic diversity (162 plant species) is very less as 1. Manhas, R.K., Kandwal, Manish K., Dhyani,
compared to Mothronwala swamp (Sharma and Joshi, Sandeep, Singh, Lokender and Joshi, S.P. (2007)
2008), and higher than Karwapani swamp (Dhyani and Effect of Soil Moisture on Demographic
Joshi, 2007). The possible reason for less floristic Dispersion, Species Association and Diversity
diversity may be closeness of swamp to human of Primary Producers in a Sub–Tropical Swamp
habitations and dependence of human population on Forest. Indian Forester, 133 (4): 547-560.
these swamp forests for fuelwood, fodder, food, 2. Kanjilal, U.N. (1901) Swamp forest in D. dun,
medicinal plants etc. Sharma and Joshi (2008) have N.W. Province. Indian Forester, 27: 228-30.
also given similar reasons for the dwindling diversity 3. Dakshini, K.M.M. (1960a) The vegetation of
and degradation of Mothronwala swamp of Doon Mothronwala Swamp forest, A preliminary
valley. We found that herbs were the most dominant Survey. Bulletin of Botanical Suvvey of India, 9:
habit followed by shrubs among all the plant forms. 57-59.
Sharma and Joshi (2008) have also reported similar 4. Dakshini, K.M.M (1960b) The vegetation of
results from Mothronwala swamp of Doon valley. Mothronwala swamp forest (Plant communities
Dominance of herbs and shrubs again signify high rate of swamp zone). Indian Forester, 86: 728-733.
of anthropogenic disturbances. 5. Dakshini, K.M.M (1965) A study of the
vegetation of Mothronwala Swamp forest
High percentage of therophytes in the present Dehradun, India. Journal of Indian Botanical
study is an indicator of the amount of influence such as Society, 44: 441-448.
grazing (Tiwari, 2005) and anthropogenic activities 6. Dakshini, K.M.M. (1970) The flora of
like catching of fishes and other eatable fauna, Mothronwala Swamp. Journal of Bombay
collection of vegetables etc. (Manhas et al., 2007), Natural History Society, 67:176-186.
which maintain the vegetation open for further invasion 7. Dakshini, K.M.M. (1974) The flora of
of therophytes. The dominance of therophytes also Mothronwala Swamp. Journal of Bombay
point towards the harsh environmental conditions of Natural History Society, 71(2): 235-243.
the swamp, which provide very limited niche space to 8. Dhyani, S. and Joshi, S.P. (2007) Angiospermic
vegetation of these marshy areas. diversity of Karwapani fresh water swamp forest
in Doon valley, India. Indian Forester, 133 (8):
ACKNOWLEDGEMENTS 1101-1108.
The authors are very thankful to the Forest Research 9. Sharma, Neelam and Joshi, S. P. (2008)
Institute and Botanical Survey of India, Northern Circle, Comparative study of a Fresh water Swamp of
Dehradun for the herbarium facilities. Doon valley. The Journal of American Science,
4(1): 7-10.
Correspondence to: 10. Som Deva and Srivastava, M.M. (1978) An
Dr. R. K. Manhas ecological study of the vegetation of Golatappar
Department of Botany, Swamp, Dehradun. Indian Journal of Forestry,
Sri Pratap College, 1(1): 44–52.
M.A. Road, Srinagar (J and K)- 190 001, India. 11. Srivastava, M. M., Joshi, S.P., Ghildiyal, J.C.,
Manhas, R. K. and Kumar, Vinod. (2000)
Demography and successional trends in Fresh

6
Marsland Press
Journal of American Science 2009: 5(1), 1-7
_____________________________________________________________________________________________
water swamps of Dehra Dun Valley (UP) India.
In: Landmarks of Botany in India (eds.: P. Soni
and N.R. Dasturiya). Surya Publications, D. Dun:
pp 113–123.
12. Kandwal, Manish K., Manhas, R.K., Joshi, S.P.
and Singh, Lokender. (2007). Phytosociological
studies of herbaceous layer in a fresh water
swamp at Golatappar, Dehra Dun. Annals of
Forestry, 15 (1): 47-61.
13. Jain and Rao (1977) A Handbook of field and
Herbarium methods. Today and Tomorrow’s
printers and Publishers, New Delhi.
14. Babu, C.R. (1977) Herbaceous Flora of Dehra
Dun. Publication and Information Directorate,
CSIR, New Delhi.
15. Raunkiaer, C. (1934) The life forms of plants
and statistical plant geography. Clarendon Press,
Oxford.

7
Marsland Press
Journal of American Science 2009: 5(1), 8-12
________________________________________________________________________
HISTOLOGICAL INTERACTIONS OF PAECILOMYCES LILACINUS
AND MELOIDOGYNE INCOGNITA ON BITTER GOURD
1
Mohd Yaqub Bhat, 2Hissa Muddin, 3Nazir Ahmad Bhat
1
Deptt. of Botany, University of Kashmir, Srinagar - 190006.
2
Plant Pathology Section, Aligarh Muslim University, Aligarh -202002, India.
3
A.P.S.U.Rewa, M.P. India
E-mail: myaqub35@gmail.com
_____________________________________________________________________________________
ABSTRACT: Momordica charantia roots were histologically examined for the interaction of
Meloidogyne incognita and the fungus Paecilomyces lilacinus which was applied at different time
intervals. Meloidogyne incognita induced large sized galls on the plants which were treated with
P.lilacinus. Fully mature females were found associated with giant cells. All the mature females on the
roots of untreated, Meloidogyne incoginta infected plants laid egg masses. The xylem and the phloem
exhibited abnormalities in structure near the giant cells.Abnormal vessel elements were occupying
larger area near giant cells. The fungus P.lilacinus. Soon after the application, entered the roots and
spread through the lumen of the vessel elements. The plants that were treated with fungus either one
weak before nematode inoculation or simultaneously, produced significantly (P= 0.01) small sized
galls in comparison to untreated plants. The size of galls remained unchanged after completion of one
life cycle by the nematode. In fungus treated plants the giant cells were small sized and the abnormality
of vascular plants was less. Paecilomyces lilacinus entered the giant cells and also into the body of
mature females. It destroyed the eggs and egg masses in and out side females. The fungus by destroying
eggs checked the possibility of secondary infection that ultimately arrested increase of gall size.
[Journal of American Science 2009: 5(1), 8-12] (ISSN: 1545-1003)

Key words: Abnormality, gall, giant cell, histology, vascular tissue.

1. INTRODUCTION (Cabanillas et al.,1988).The effects of fungus on

The fungus Paecilomyces lilacinus (Thom) M.incognita parasitizing the roots of Momordica
Samson has been reported as a potential biological charantia has not yet been reported.The objectives
control agent for root –knot nematode and other plant of this study were : (i) to examine the effect of
parasitic nematodes (jatala, et al., 1979, 1982, 1986; P.lilacinus on M.incognita infected plant tissues.(ii)
Adiko,1984;Cardona and Leguizamon,1997 and to know the effect of P.lilacinus on nematode
Khan and Williams ,1998).P. lilacinus is common development, (iii) to determine the effect of
soil hypomycete closely related to Penicillium P.lilacinus on eggs and egg masses,(iv) to examine
(Samson,1975).The sedentary stages of the root - the effect of P.lilacinus on the giant cells and (v) to
knot and cyst nematode are most vulnerable to P. evaluate the efficacy of P.lilacinus in controlling the
lilacinus. The fungus is capable of colonising disease development, on applying the fungus at
nematode reproductive structures thus causing varying time intervals, before or after nematode
destruction of females, cysts and eggs. (Franco, et al,. inoculation.
1981, Jatala, 1982, 1986; Gintis, et al., 1983 and
Cardona and Leguizamon,1997). 2. MATERIALS AND METHODS
Paecilomyces lilacinus increased the yield of Nematode cultures of Meloidogyne
tomato and okra and lowered the population of M. incognita was maintained from single egg mass on
incognita juveniles, at the mid and at the beginning egg plant (Solanum melongena L.)grown in green
of the next season in treated pots than in untreated house in 15 cm diameter earthen pots containing a
pots(Neo and Sasser 1984). M.incognita juveniles mixture of steam sterilized soil and sand (3:1).
when exposed to fungus resulted in reduced gall M.incognita originally was isolated from vegetable
formation and egg mass production. crop fields of Aligarh. The root-knot nematode was
The association of P.lilacinus with the eggs identified on the basis of characteristic perineal
of M.incognita is well documented but the exact pattern and North Carolina differential host test.
mode of its parasitism is unknown. Root galling and Freshly hatched second stage juvenile inoculum
giant cell formation were absent in tomato roots was prepared by hatching egg masses picked from
inoculated with fungus infected eggs of M.incognita. egg plant roots, maintained as pure culture in green
P. lilacinus colonised surface of epidermal cells as house. Paecilomyces lilacinus, used in the experiment,
well as the internal cells of cortex of tomato roots was obtained from international potato centre, Lima,

8
Histological Interactions Of Paecilomyces Lilacinus And Meloidogyne Incognita On Bitter Gourd
Mohd Yaqub Bhat et al.
_________________________________________________________________________________________________________________

Peru. The fungus was cultured on PDA for 15 days at 27±20C,

then inoculated to Richards Medium (Riker and Riker, 4) T4- inoculated with 1,000 J2 and treated with
1936) for en-masse propagation .The mycelia (100 fungus simultaneously
gm) were blended in distilled water (100 ml) in 5) T5- inoculated with 1,000 J2 and treated with
warring blender to make mycelial suspension for soil fungus two week after inoculation
application (10 ml of suspension containing 1gm Each treatment was replicated five times, arranged in
mycelia).The fungus was applied into the rhizosphere randomised block design. Each set of plants was
zone by making three or four holes around the plant. uprooted carefully after 45days . The roots were cut
Plant Materials : Seeds of Momordica into small pieces and processed for
charantia L. variety NSC were surface sterilised histopathological studies. Root pieces were fixed in
with 1% sodium hypochlorite (NaOCl) for five formalin- aceto - alcohol (F.A.A) and then
minutes and rinsed three times with sterile distilled dehydrated through tertiary-butyl-alcohol (T.B.A.)
water. 100-150 axenised seeds were placed on a schedule as given by Johansen (1940). Infiltration
moist sterilised filter paper kept in sterilised petri and embedding of root pieces in paraffin wax was
dishes. Seeds were allowed to germinate. The followed and sections of 12ȝm thickness were taken
germinated seeds were transferred to 15 cm diameter with the help of rotary microtome in the form of
clay pots filled with autoclaved soil, sand and ribbon. The ribbons were cut into small pieces and
farmyard manure in the ratio of 7:3:1. Two week old mounted on clean slides with the help of Haupt’s
seedlings were inoculated with a suspension of 1000 adhesive and 3% formalin and stained with safranin
J2 pipetted into root zone via the holes in the and fast green (Sass1951). The slides were taken out
rhizosphere zone around plant in each pot. To from xylene. The mounting medium was applied on
achieve the aim and objective the experiment was the surface of slide before evaporation of xylene
designed as per the following treatment schedule and cover slip was lowered gradually. Finished slides
1) T1- un-inoculated control were left at room temperature at least for 24 hours
2) T2- inoculated with 1,000 J2 only and then kept in an incubator at 400C.The slides were
3) T3- inoculated with 1,000 J2 and treated with examined under light microscope and necessary
fungus one week before inoculation photographs were taken.

Fig 1: Showing heavy infestation of Meloidogyne Fig 2: Showing normal xylem (NX) strands
incognita (N). The mature females have egg masses (EM). with conidiophores and conidial chain
Abnormal xylem (AX) and abnormal phloem (AP) C in the lumen of vessel elements.
are in abundance.(25X)

9
Marsland Press
Journal of American Science 2009: 5(1), 8-12
________________________________________________________________________

Fig 3: Showing abundant growth of Peacilomyces Fig 4: Showing hyphae (H) and conidial chains
lilacinus hyphae (H) in and around the females (C) at the root surface. (12.5 X)
of M. incognita (N).(32 X).

3. RESULTS xylem and phloem were present in Paecilomyces

Histologically the primary root of lilacinus treated plants. The abnormality in vascular
Momordica charantia consists of uniseriate elements was less. The fungal hyphae destroyed
epidermis, multilayered parenchymatous cortex and eggs and egg masses and also entered into the body
stele. The stele in primary root may be diarch, triarch of the females. Around the nematode body fungal
but generally tetrarch, displaying a typical growth was abundant (Fig 3).The root surface also
dicotyledonous pattern. Xylem and phloem are exhibited profused growth of fungus (Fig 4). The
radially arranged alternating with one another. There simultaneous application of root knot nematode and
is small pith consisting of parenchyma cells at the Paecilomyces lilacinus not only destroyed eggs and
centre of four xylem arches. Meloidogyne incognita egg masses, but also entered into the internal tissues
inoculated M.charantia plants receiving no disease of root, either intercellularly or intracellularly as is
controlling treatment produced large sized galls. The evident from the transverse sections of vessel
mature females feeding on giant cells, abnormal elements (Fig 5).The egg masses were destroyed by
xylem and abnormal phloem was observed (Fig the fungus and the growth of fungus was profound
1).All the mature females were found associated with inside egg masses. There was no change in the size
egg masses. Histological examination of the galled of giant cells and amount of vascular elements as
roots which were given Paecilomyces lilacinus compared to untreated plants. In plants treated with
treatment, one week before the nematode inoculation, Paecilomyces lilacinus, one week after nematode
revealed that the fungus entered the root tissue and inoculation, the fungal hyphae was observed inside
grew successfully. The hyphae and conidiophores, the giant cells (Fig 6).The fungal growth was profuse
bearing chains of conidia were seen in normal around the body of developing nematodes. In the
vessel elements of the xylem (Fig 2).The giant cells, normal tissue the fungus spreads both inter and
though smaller, resembled with those of T2 intracellularly as is evident from figure (Fig.7)
plants .In the vicinity of the giant cells abnormal

Fig 5: Showing vessel elements (VE) in Fig 6: Showing hyphae (H) in giant cell (GC)
transverse sections enclosing conidial and nematode (N) adjacent to giant
chains (C).(40 X) cell.(25 X)

10
Histological Interactions Of Paecilomyces Lilacinus And Meloidogyne Incognita On Bitter Gourd
Mohd Yaqub Bhat et al.
_________________________________________________________________________________________________________________

Fig 7: Showing normal xylem (NX) strands with hyphae(H) of Paecilomyces lilacinus
traversing through the lumen of vessel elements. (32 X)

DISCUSSION
The fungus Paecilomyces lilacinus shows abnormal xylem and phloem indicated that the
diverse modes of habits. Basically it is a saprophyte nematode activity and development was influenced
(Domsch et al., 1980) and can easily be grown on by the presence of P.lilacinus. Large giant cells and
artificial culture media. It behaves as an epiphyte and more quantity of abnormal tissues showed that the
grows on the surface of plant roots (Cabanillas nematodes which entered earlier were not affected by
et.al.,1988). It also grows inside the root tissue and the fungus. On the basis of these observations we
behaves as an endophyte and does not cause any concluded that the fungus can not check primary
damage to the plant. Still at other times it parasitizes infection of nematode when the plants are attacked by
eggs and egg masses of Meloidogyne species and the juveniles. However, it can check secondary
destroy them. Because of the lastly stated behaviour, infection because it destroys eggs as and when these
P.lilacinus has been used by several workers, as are deposited. As far as time of application of
biocontrol agent against root-knot nematode and P.lilacinus is concerned, from our studies it can be
other nematodes (Jatala, et al., 1979). suggested that incorporation of fungus P.lilacinus
Paecilomyces lilacinus was encountered one week before and at the time of nematode
frequently in and around normal and abnormal inoculation is more effective in controlling the root-
xylem. In our opinion, vessels and vessel elements knot disease, as compared to later intervals of
provide sufficient space for its development and also fungus applications.
provide an uninterupted passage to grow inside the
plant tissues. We consider that P.lilacinus develops ACKNOWLEDGEMENT
inside the root tissues inter and intracellularly. The authors are thankful to Chairman, Department of
Whether the fungus is beneficial or not to plant but, it Botany, Aligarh Muslim University, Aligarh for
is not harmful. In all the sections studied, the fungus providing necessary facilities to complete this work.
was not found damaging the plant tissues even when
it was in abundance. Further it did not affect the giant REFERENCE
cells in which its occurrence was noted. In all the
treatments it was regularly observed that P.lilacinus 1. Adiko, A., 1984. Biological control of
damage the eggs and egg masses. Various workers Meloidogyne incognita with Paecilomyces
have reported egg destroying activity of this fungus lilacinus. M.S. Thesis , Department of plant
(Jatala, et al,. 1979, Jatala, 1985,1986.) It has also pathology, North Carolina State University ,
been reported that the fungus can destroy neither the Raleigh,
juveniles nor the adult females (Jatala ,1986). The 2. Cabanillas E., Barker, K.R. and Daykin, M.I.
eggs however seem to be the most preferred target for 1988. Histology of the interactions of
obtaining the nourishment by the fungus. Contrary to Paecilomyces lilacinus with Meloidogyne
this (Cardona and Leguizamon,1997 ) reported 94% incognita on tomato. J. Nematol. 20: 362-265.
infection in Meloidogyne spp. by P. lilacinus strain- 3. Cardona, B.N.L. and Leguizamon, C. J. E. 1997.
9207. KHAN and WILLIAMS (1998) found Isolation and pathogencity of fungi and bacteria
P.lilacinus entering into the body of the females to the root-knot nematode of coffee
through natural opening. They did not mention Meloidogyne spp. Goldi. Fitopatologia
whether the fungus damaged the females or not, Colombena . 21: 39-52,
although it damaged the eggs inside the egg masses.
Small sized giant cells and small amount of

11
Marsland Press
Journal of American Science 2009: 5(1), 8-12
________________________________________________________________________
4. Domsch, K.H., Gams, W. and Anderson, T. H. 9. Jatala, P.1986. Biological control of plant
1980 .Compendium of soil fungi. Vol. 1: parasitic nematodes. - Ann. Rev. Phytopath. 24:
Academic press, New york, pp.859. 453-489
5. Franco, J., P. Jatala, and . Bocangel, B. 1981. 10. Jatala, P., Kaltenback, R. and Bocangel,. M.,
Efficacy of Paecilomyces lilacinus as biocontrol 1979. Biological control of Meloidogyne acrita
agent on Globodera pallida.. J. Nematol. 13: 438- and Globodera pallida on potatoes. J. Nematol.
439. 11: 303.
6. Gintis, B.O., Morgan-jones, G. and Rodriguez- 11. Johansen, D.A.1940. In Plant Microtechnique
Kabana. R. 1983. Fungi associated with several Mc Graw Hill, New Book Co. New York, 523 pp.
developmental stages of Heterodera glycines 12. Khan, A. and K. L. Williams, 1998. Recent
from an Albama soybean field soil. studies on Paecilomyces lilacinus as
Nematropica .13: 181-200. bionematicide. Nematologica 44: 519-520.
7. Jatala, P.,1982. Biological control with fungus 13. Neo, J.P and Sasser, J. N. 1985. Evaluation of
Paecilomyces lilacinus. Progress to date and Paecilomyces lilacinus as an agent for reducing
possibilities for collaborative research between yield losses due to Meloidogyne incognita.
CIP and IMP. Collaborators pp.214-218. In Biocontrol. 1: 57-67.
proceedings to third research conference on root 14. Riker, A.J and Riker, R.S., 1936. Introduction to
knot nematodes Meloidogyne spp.22-26 research on plant disease. St. Louis and New
March,1982,Region 2,Raleigh;North Carolina York, John S, Swift Co., 117p.
State University , Graphics. 15. Samson, R.A. 1975. Paecilomyces and some
8. Jatala, P. 1985. Biological control of nematodes. allied hypomycetes. Studies in Mycology 6: 1-
p 301-303. - In advanced Treatise on 119.
Meloidogyne Vol.1: Biology and Control Eds. 16. Sass. J.E.1951. In Botanical Microtechnique,
J.N. Sasser and C.C Carter. A cooperative Iowa College Press, Ames, Iowa, 228 pp.
publication of the department of Plant Pathology
and U.S.AID, N.C.S.U. Graphics pp.422.

12
Marsland Press
Journal of American Science 2009:5(1) 13-18
______________________________________________________________________________________________
Experimental Study on The Dynamic Behaviors of the Material for Clay core
wall sand dams

Xu Shangjie1,2,Dang Faning1,TianWei1,ChengMo2
1.Xi’an University of Technology, Xi’an, Shanxi, 710048,China;
2.Shandong Provincial Institute of Water Resources, Ji’nan, Shandong, 250013,China
[XVKDQJMLH#VLQDFRP
________________________________________________________________________________________
Abstract˖The Clay core wall sand dams are mostly constructed on the tremendously thick covering layers
with the base built with gravelly coarse sand and the upstream and downstream built with sanded shell into
which gravelly coarse sand by manpower. The structures of these dams are generally loose and are presumably
thereby considered not to meet the requirements against earthquake. In order to perform further safety analysis
and liquefaction judgment with these dams in relation to their anti-seismic capacity,a typical dam with clay
core and sanded shell was chosen and experiments were carried out to observe the dynamic behaviors of the
dam , including the dynamic strength, the dynamic deformation and the dynamic pore pressure of the dam base
and its upstream sanded shell. The results showed that the vibration stress ratio of gravelly coarse sand
decreased with the increment in vibration frequency and increased with the increment in consolidation ratio,
that the elastic modulus of gravelly coarse sand decreased with the increment in strain, and that the damping
ratio of gravelly coarse sand increased with the increment in strain. In conclusion, the denser the dam material
is, the better the anti- seismic behavior is, the sand used in the dam is non-linear in nature, The model of
vibration pore water pressure growth is characterized by simplicity in expression, convenience in application,
and being able to used in widespread way, etc. It reveals what inherent in the relationship of the increment of
residual pore pressure with multiple factors, and hence can be used in the dynamic analysis of effective stress.
[Journal of American Science 2009:5(1) 13-18](ISSN: 1545-1003).

Key words˖gravelly coarse sand, earthquake liquefaction, dynamic behaviors, upstream sanded shell

1 Introduction their leading to catastrophic outcomes brought by

More than 90% of reservoirs were constructed possibly occurring seism in the future while leaving
during the period of the so-called “Great leap forward” them there for multiple usages. In this study, the
around the year of 1958 as some kind of special materials from the dam for Houlonghe reservoir were
products with most of the dams being of clay-core- chosen to observe experimentally their dynamic
sand-shell type. And the filling of these dams were properties with respect to the anti-seismic capacity of
accomplished using the so-called “tactics of human sea” the present dam made from them. The dam for
with their internal structures comparatively loosely built Houlonghe reservoir is located on the middle stream of
with no consideration into their capacity against seism the east branch of Gu river at Yatou town, Rongcheng
at all. Consequently, these dams have been presumed city, Shandong province, China, which is a typical
not to be able to reach anti-seismic requirements from clay-core-sand-sand-shell dam. This dam controls water
the beginning. Even worse, these dams have undergone flow of about 61km2. And the reservoir with a total
operation of 50 years with severe problems associated capacity of about 5.3 millionm3, is a important medium
with aging. Therefore, it has been extremely critical to sized one of its kind and comprehensively applied for
evaluate the real property of anti-seism of these dams so flood prevention and water supply. The dam was
as to perform reasonable measures for improving their constructed in 1959 and has affiliated with aging
anti-seismic capacity , and thus reducing probability of diseases seriously after undergoing operation of 50

13
Experimental Study on The Dynamic Behaviors of the Material for Clay core wall sand dams
Xu Shangjie,Dang Faning,TianWei,Cheng Mo
______________________________________________________________________________________________
years. In 2004, it was listed as dangerous one after characteristics of gravelly coarse sand in order to
safety assessment by the national security department provide theoretical basis for related institutions
for reservoir and dam, and thus needs danger control involving in the analysis of the dynamic safety of the
and reinforcement. dam and supply data for the evaluation of seismic
The dam is a clay-core-shell-wall dam with its liquefaction.
upstream being sanded shell and its base made from 2 Samples for the experimentation
gravelly coarse sand. The reservoir has been set to be in The experimental soil material was the loose soil
defense against earthquake of seven degrees by Richter obtained from the typical sections of sanded shell in the
scale. But the judgment has been made through upstream and the sand layer at the base of the dam for
reconnaissance that there has been existing the Houlonghe reservoir. The characteristics of the two
probability of seismic liquefaction with the sanded shell soils were shown in table 1. The dry density of them:
in the upstream of the dam and the sanded layer in the the soil of the upstream sanded shell: Ȗd=1.62g/cm3, the
base of it in the circumstance of earthquake. Therefore, soil of the sand layer at dam base:Ȗd=1.58 g/cm3.
we carried out the research to study the dynamic
Table1.
the characteristics of soils obtained from sanded shell in the upstream and sand layer at the base
sampling particle size (mm)/%
sampling nonuniform curvature Name in
depth 2̚ 0.5̚ 0.25̚
positions >2 <0.075 coefficient coefficient door
˄m˅ 0.5 0.25 0.075
upstream sanded gravelly
3.0 13.7 29.1 32.4 19.0 5.8 5.91 1.41
shell coarse sand
the sand layer at gravelly
1.5 22.6 33.2 28.2 12.8 3.2 9.73 1.08
dam base coarse sand
3 METHODS OF EXPERIMENTATION
The DSZ-100 electromagnetic vibration triaxial and damp ratio: Dynamic elastic modulus (Ed) is the
ratio of dynamic-stress ıd and dynamic-strain İd , which
apparatus was employed for the observation of the reflects the relationship between dynamic-stress and
seismic behavior of the material of the dam. The sample dynamic-strain during the phase of modification in
shape under the action of periodically loading. Damp
was 3.91cm in diameter, 8cm in high, and greater than ratio Ȝ is the ratio of damp coefficient c and critical
damp coefficient ccr. , measured by cyclic triaxial test,
90% in saturation. Three consolidation ratios were used indicating the energy-dissipation per vibrating cycle,
in this experiment: Kc=1.0,1.5,2.0. The consolidation and is therefore also called the equivalent gummy damp
ratio of soil. The method used for examining
stresses were 0.05MPa,0.10MPa,0.15MPa, respectively. dynamic-elasitc modulus and damp ratio in this study
Sine wave with 1Hz in frequency was used with was as follows: the samples were consolidated with
different stresses , and then the samples were
dynamic wave is. respectively implemented with dynamic stresses from
Method for liquefaction experiment: The tested low grade to high grade progressively in step with 10
times per graded cycle in under the circumstance of
sample was prepared in such way that a given amount non-draining. The resultant hydraulic pressure in the
of dried sand by baking was mixed with water,and then accessory small openings in the samples was dissipated
after the termination of every graded loading before the
boiled to be dried after thorough stirring. The process of implementation of the next graded loading in order to
keep the valid stress constant during the process of the
consolidation of the sample was accomplished by test.
applying axial stress ı1 and lateral stressı3 to the The criteria for demolition examination at equal
consolidation was set at 5% of the peak of the axial
sample under desired conditions and then the strain ( double peaked value). The criteria for
liquefaction test was performed by applying given axial demolition examination at non-equal consolidation was
also set at 5% of the axial halved peak value plus
circulation stress in the circumstance of no draining. residual deformation.
Method for measuring dynamic elastic modulus

14
Marsland Press
Journal of American Science 2009:5(1) 13-18
______________________________________________________________________________________________
4 DYNAMIC CHARACTERISTICS OF THE of circulations at 10, in viewing of that the earthquake
MATERIALS OF THE DAM defence intensity had been set at seven degrees by
4.1 The characteristics of Dynamic Intensity Richter scale for the dam of Houlonghe reservoir. The
Dynamic intensity refers to the dynamic shearing ratios of liquefaction-stress when Nf at 10 were shown
stress leading to sample failure with a given number of in table 2, and the relationship curve of the dynamic
circulating vibration force, Nf. The relationship curve, shearing stress ratio ıd/(2ı3) of upstream sanded shell
ıd/(2ı3) ̚ Nf was displayed on uni-logarithm grapy and the number of vibration destruction, Nf was shown
sheet base on the results of the test. The curve was in figure 1.
established with the results of the test when the number
Table2.
liquefaction- stress ratio of the materials of the dam
Liquefaction- stress ratio³G³
sampling positions
  

upstream sanded shell   

the sand layer at dam base   

Fig 1. The relationship curve of ıd/(2ı3)

The values of dynamic stresses at different dynamic strength parameters, ĭd and Cd at three
consolidation stresses were obtained from liquefaction- different consolidation stresses were shown in table 3.
stress ratios when the times of circulating force at 10, The Mohr circles of upstream sanded shell of the dam at
and thus then the Mohr circle could be drawn for the different stresses were shown in figure 2.
determination of the dynamic strength parameters. The
Table3.
dynamic strength parameters at three different consolidation stresses of the materials of the dam

1.0 2.0 3.0

sampling positions žd Cd žd Cd žd Cd
(degree) (MPa) (degree) (MPa) (degree) (MPa)

upstream sanded shell 14.81 0 25.69 0 30.55 0

the sand layer at dam base 14.2 0 24.6 0 30.92 0

15
Experimental Study on The Dynamic Behaviors of the Material for Clay core wall sand dams
Xu Shangjie,Dang Faning,TianWei,Cheng Mo
______________________________________________________________________________________________

Fig 2. The Mohr circles of upstream sanded shell at different stresses(n=10).

In order to perform the liquefaction analysis, the The shearing stress during liquefaction cycle, IJfd ,
liquefaction shearing stress, IJfd , under different static resulted from the combination of a certain initial
stresses, ıfs, had to be determined. The curve effective directed stress ıfs and the initial shearing stress
representing relationship of initial shearing stress ratio, IJfs(e.g. anti-liquefaction shear stress)could be obtained
ȕ=IJfs/ıf , with Į=IJfd/ıfs were obtained from the from the curve.
results of the test as shown in figure 3.
Į
0.4

0.3

0.2

0.1

0 ǃ
0.1 0.2 0.3 0.4 0.5

Fig 3. The relationship of Į and ȕ in upstream sanded shell

4.2 The characteristics of dynamic deformation was determined by the following equation.
The relationship between the maximal dynamic n
§ V 0/ ·
elastic modulus Edmax and the averaged effective E d max kPa ¨¨ ¸¸ ˄1˅
© Pa ¹
consolidation pressure V 0/ was determined , and the
In the equation, V 0/ is the averaged main
relationship curve of the elastic modulus Ed, and damp
ratio Ȝd against dynamic strainİd was established for effective stress when samples being consolidated,
the determination of the characteristics of dynamic
which is determined by the equation: V 0/ =
deformation.
According to the results of the test, the / /
˄ V 1 + 2V 3 ˅/3˗ k and n are test constants relative to
relationship of maximal dynamic elastic modulus Edmax
properties of soils used in tests. Pa is the atmospheric
and the averaged effective consolidation stress V 0/
pressure.

16
Marsland Press
Journal of American Science 2009:5(1) 13-18
______________________________________________________________________________________________
/ coordination, and N was the slope ratio. The values of k
Edmax̚ V 0 relationship curve was drawn on the
and n for the materials of the dam when consolidation
double logarithm coordination sheet, wherein K was the ratio at 1 were shown in table 4.
intercept of the curve on the vertical axial of the
Table4 k and n with the material of the dam
the material of the dam k n

upstream sanded shell 40 0.681

the sand layer at dam base 38.5 0.635

The dynamic elastic modulus Ed decreased with The curve for the relationship between damp ratio
the increasing of axial strain while it increased with the Ȝd and dynamic strain İd was depicted on the halved
increasing of the consolidation stress ratio. Nevertheless, logarithm coordination sheet. the half logarithm squared
the ratio of modulus Ed and maximum of dynamic paper. The following equation was obtained after
elastic modulus Edmax , Ed/Edmax did not change approximation of the curve for the Ȝd̚İd relationship:
significantly with the alteration of consolidation Ȝd= aİd/(b+İd) (3)
pressure. The relationship of Ed/Edmax and İd were In the equation, a and b are the approximation
depicted on the halved logarithm coordination sheet parameter.
based on the results of the test. The following equation The relationship curve of Ed/EdmaxǃȜd andİd with
was obtained after approximation of the curve for the shell of the upstream of the dam was showed in
Ed/Edmax ̚İd . figure 4. The approximation parameter for elastic
Ed/Edmax =1/(1 +İd/w) (2) modulus w with dam material and the approximation
In the equation, w is the approximation parameters for damp ratio, a and b were shown in table
parameter. 5.

Fig 4. The respectively corresponding relationshipof Ed/EdmaxǃȜdandİdin upstream sanded shell

Table 5.
Approximation parameterof elastic modulus damp ratio
the material of the dam w a b
upstream sanded shell 0.001596 0.4767 0.001
the sand layer at dam base 0.001303 0.2223 0.006

4.3 THE CHARACTERISTICS OF DYNAMIC During the vibrating triaxial test , the changes in
PORE PRESSURE small opening hydraulic pressure were recorded

17
Experimental Study on The Dynamic Behaviors of the Material for Clay core wall sand dams
Xu Shangjie,Dang Faning,TianWei,Cheng Mo
______________________________________________________________________________________________
simultaneously with computer and thus the curve for straight line, indicating that the results were
comparatively is reasonable. The results also showed
the relationship of changes in small opening hydraulic
that the stress ratio of each kind of dam material
pressure with times of vibration was obtained. Data of increased with the increment of consolidation ratio and
the test was arranged with the vibration pore water the dynamic strength index increased significantly with
pressure growth model proposed by the Institute of the increment of consolidation ratio, indicating that the
denser the dam material is, the better the anti- seismic
Water Resources, the Yellow River Committee, behavior is.
China ,which is denoted in the following exponential (2) The results in the dynamic modulus and damp
function. measurements showed that the dynamic elastic modulus
of the dam material decreased with the increment of the
U 1  (1  U 0 )10  K[ /(1[ ) ˄4˅ strain and damp ratio increased with the increment of
the strain, indicating that the sand used in the dam is
In the equation, ȟ is the ratio of the non-linear in nature. The strain ranged from 10- 4 to 10- 2
in this test, in which modulus ratio and damp ratio
destruction-to-times in logarithm, which could be
were the actual values. The values of the modulus ratio
obtained by the equation, [ log N / log N f ; U0 is and damp ratio in other ranges could be obtained
through simulation curve.
the relative pore pressure ratio of the first cycle. (3) The model of vibration pore water pressure
growth is characterized by simplicity in expression,
The test constants k was determined by the convenience in application, and being able to used in
following equation. widespread way, etc. It reveals what inherent in the
relationship of the increment of residual pore pressure
K DN f E (5) with multiple factors, and hence can be used in the
dynamic analysis of effective stress.
U0 was determined by the following equation.
REFERENCES:
U 0 JN f T (6) 1. YUAN Ju-yun, XU Chao, ZHAO Chun-feng. Soil
lab test and in-situ testing[M].Shanghai: Tongji
Nf was determined by the following equation. University Press, 2004.
B 2. Institute of Hydraulic Research,YRCC, Study on
D d AN f ˄7˅ the anti-seismic properties ofSand and Gravel in
Xiaolangdi earth rockfill dam,(national technology
WhereinĮd was the ratio of dynamil stress, which
tacking report of 8th five-year plan) ,1994.
was determined by the equation, Įd=ıd/(2ı3). Thus, the
3. Shandong Provincial Institute of Water
pore pressure of any time vibration was determined by
Resources, Engineering Geological Investigation of
the following 7 constants, namely, AǃBǃȖǃșǃĮǃ
Houlonghe reservoir in Rongcheng city.
ȕ and uf, where uf was the averaged value of pore
4. XU Bin, KONG Xian-jing, ZOU De-gao, Study of
pressure ratio with predetermined destruction standard
dynamic pore water pressure and axial strain in
in the same consolidation ratio .
saturated sand-gravel composites[J]ˈRock and Soil
Different consolidation ratio and consolidation
Mechanicsˈ2006,27(6).
pressure has taken into consideration in this model,
5. SUN Jing, YUAN Xiao-ming, SUN Rui.
which is thus characterized by simplicity in expression,
Reasonabilitycomparison between recommended
convenience in application, and being able to used in
and code values of dynamic shear modulus and
widespread way, etc. It revealed what inherent in the
damping ratio of soils
relationship of the increment of residual pore pressure
6. Earthquake Engineering and Engineering
with multiple factors, and hence could be used in the
Vibration,2004, 24(2): 125ˉ133.
dynamic analysis of effective stress.
7. BORDEN R H, SHAO L, GUPTA A. Dynamic
5 Conclusion
propertiesof piedmont residual soils[J]. Journal of
(1) The results showed in the present test with
Geotechnical and Geoenvironmental Engineering,
dynamic strength that the vibration-stress ratio of the
ASCE, 1996,
material of the dam increased with the reduction of
8. TOWHATA I, ISHIHARA K. Undrained strength
times of vibration, and the numerical points of stress
of sand undergoing cyclic rotation of principal
ratio at different confining pressure fell relatively well
stress axes[J].
into a narrowly-defined band under the same
9. Soils and Foundations, JSSMFE, 1985, 25(2)
consolidation ratio and thus could be expressed as a

18
Marsland Press
Journal of American 2009: 5(1), 19-28
______________________________________________________________________________________
Research Article
Influence of Subchronic Exposure of Profenofos on Biochemical
Markers and Microelements in Testicular Tissue of Rats

Afaf A. El-Kashoury
Department Of Mammalian And Aquatic Toxicology, Central Agricultural
Pesticides Laboratory, Agricultural Research Center, Giza, Egypt
____________________________________________________________________________
ABSTRACT: To investigate the effect following subchronic exposure to the organophosphorous
insecticide of common name profenofos, which extensively used in agriculture, on the key
enzymes of fertility and the concentration of microelements in testicular tissues in male albino rats.
Methods: Adult male albino rats were orally administered with profenofos at a does of 23.14
mg/kg body weight per day for 60 days, emulsifying in 0.4 ml tap water. The control group
received equal volume of tap water. Twenty-four hours after the last treatment the rats were
sacrificed using anesthetic ether. Epididymus and testes were collected, cleaned and weight. Then
epididymus prepared in buffer saline and spermatozoa were examined with light microscopy for
concentration and motility. Testes were fractionated and supernatant of testicular homogenate was
obtained by centrifugation, activities of alkaline and acid phosphatases, lactate dehydrogenase and
total protein as well as concentration of microelements; Copper, Iron, Zinc and Selenium were
measured. Moreover, the testes were histologically examined. Results: The epididymus and testes
weights were significantly decreased. Reduction in sperm count was recorded in cauda
epididymus in profenofos treated group, associated with decreased motility. Total protein (TP)
level exhibited an elevation in testicular tissue in comparison with the control group. There was
significant decrease in the activities of alkaline and acid phosphatase (ALP and ACP) and lactate
dehydrogenase (LDH). A totally different trend was observed for the level of microelements;
Copper (Cu), Zinc (Zn), Iron (Fe) and selenium (Se) where a sharp augmentation in the element
levels was noticed in profenofos-treated rats compared with the control group. Treatment-
dependent histopathological changes were seen in testes. Conclusion: Profenofos alters testicular
functions possible by inhibition the activities of marker enzymes and inducing alteration in
microelements levels, thereby disrupting male reproduction. [Journal of American 2009: 5(1), 19-
28] (ISSN: 1545-1003)

Key words: Profenofos, Lactate dehydrogenase

1. INTRODUCTION disorders, environment or occupational exposure

Organophosphorous insecticides (OPIs) have been to chemical or other means. Specific causes of
considered as genuine alternatives to chlorinated testicular damage have been catalogued
(O'Ch) insecticides due to their broad-spectrum (Jadaramkunti and Kaliwal, 2002). Mainly, much
pesticidal properties and relatively shorter data are available about biochemical analysis of
persistence after applications (Sharma et al., 2005). seminal plasma. However, not many studies have
OPIs in addition to their intended effects like been conducted in animals yet (Pesch et al., 2006).
control of insects or other pests are sometimes Analysis of enzyme activities and concentrations
found even to effect non-target organisms of microelements can estimate integrity and
including human beings (Chantelli-Forti et al., function of testes, in man; analysis of seminal
1993; Chaudhuri et al., 1999). Exposure to low plasma enzymes and microelements has been
level OPIs is known to produce a variety of performed accurately and much is known about the
biochemical changes, some of which may be importance of the "right contents" of seminal
responsible for the adverse biological effects plasma (Pandy et al., 1983; Chia et al., 2000;
reported in humans and experimental animals Huang et al., 2000 and Stanwell-Smith et al.,
(Sutatos, 1994). There is growing concern that 1983). It has been reported that, pesticides with
environmental chemicals both natural and man- such properties have been shown to cause
made, having estrogenic property may be causing overproduction of reactive oxygen species (ROS)
a variety of reproductive disorders in wildlife and in both intra and extra cellular spaces, resulting in
human population (Chitra et al., 1999). The testes a decline of sperm count and infertility in wildlife
of humans and other mammals are highly and human (Gangadharan et al., 2001).
susceptible to damage produced by genetic

19
Influence of Subchronic Exposure of Profenofos on Biochemical Markers and Microelements in Testicular Tissue of Rats
Afaf A. El-Kashoury
____________________________________________________________________________________________________________

Trace elements, such as Copper (Cu), First group: (n = 10) served as normal control and
Zinc (Zn), and Selenium (Se) have a pivotal role in animals were received the vehicle (tap water).
the spematogensis (Homma-Taked et al., 2003) Second group: (n = 20) animals were orally dosed
Ionic environment has a high influence on sperm for 60 days with profenofos at 23.14 mg/kg body
function (Hamameh and Gatti, 1998), profenofos weight (4 doses/week). Clinical signs were
belongs to the phosphorothioate class of OPIs. It monitored daily and animals were weighed twice
widely used for a variety of agricultural and public weekly throughout the experiment and the dose
health applications, previous studies suggest that was adjusted accordingly.
profenofos considered as one of the male
reproductive toxicant (Moustafa et al., 2007). In After completion of treatment period (60
spite of the extensive use of profenofos in crop days), animals were anaesthetized with ether and
protection and in the household, information sacrified. The testes and epididymus were removed
related to its effects on health with particular immediately, cleaned of the adhering tissues and
reference to reproductive toxicity are scarcely. weighted. Fertility-related parameters (sperm
Therefore, the objective of this study was to clarify count and motility) were performed by dissecting
the effect following subchronic exposure to out the Cauda epididymus and teasing it in a
profenofos on testicular functions by measuring known volume of normal saline at 370C. Sperm
the fertility indices (sperm count and motility), the counting was done using a haemocytometer
activity of specific enzymes that responsible of according to the method of Feustan et al. (1989).
spermatogenesis (alkaline and acid phosphatases The right testes were kept in a deep freezer (-400C)
and lactate dehydrogenase) and total protein level for biochemical estimations and microelements
as well as concentrations of the essential detection. Left testes were removed and fixed in 10
microelements; Copper (Cu), Iron (Fe), Zinc (Zn) % formalin for routine histopathology.
and Selenium (Se) in testicular tissue of male rats.
Frozen testes were washed with saline
2. MATERIALS AND METHODS solution, then minced and homogenized (10%
The active substance profenofos produced by W/V) in ice-cold saline, using a chilled glass-
Syngenta multi national comp. under trade name: teflon porter-Elvehjem tissue grinder tube. The
Selecron 72% EC was used. Tap water was used homogenate was centrifuged at 10,000 xg for 20
for preparing emulsion of profenofos immediately min. at 4 0C and the resultant supernatant used for
before use and orally administered into animals by determination of protein contents, Tp (Bradford,
osophageal intupation (per OS.). The median lethal 1976); alkaline phosphatase, ALP (Babson, 1965)
dose (LD50) of profenofos (per OS.) was and acid phosphatase ACP (Babson and Read,
determined according to Weil (1952) and its value 1959). Also, a 10% homogenate of testes was
was 185.13 mg/kg body weight. prepared in ice-cold 0.1M phosphate buffer, the
homogenate was centrifuged at 12,000 xg for 30
In this investigation, thirty male Wistar min. at 40C. the supernatant used for determination
albino rats, rattus norvegicus were obtained from of lactate dehydrogenase, LDH (Moss and
the breeding unit of the Egyptian organization for Henderson, 1994).
the Biology and vaccine production, Egypt. Male
rats initially weighing 150+10g were used. For the histopathological observations at
Animals were allowed to be acclimatized to light microscopic level, fresh testes were
laboratory conditions; of temperature at 25+20C, immersion fixed in 10% formalin saline.
humidity (30-70%) and light (12-h dark: 12-h Following an overnight fixation, the specimens
light) and kept on balanced diet and water ad were dehydrated in ascending grades of alcohol,
libitum for 2 weeks prior to the experiment. cleared in benzene and embedded in paraffin wax.
Animals were housed throughout the experiment Blocks were made and 5um thick sections were
in polypropylene cages (with each cage housing double stained with hematoxylin and eosin and
five animals) containing paddy husk as bedding. observed under microscope (Banchraft et al.,
2-3 Experimental Design. Rats were randomly 1996).
divided into two comparable groups as follows,

20
Marsland Press
Journal of American 2009: 5(1), 19-28
______________________________________________________________________________________

The concentrations of the microelements differences with a value of p<0.05 were considered
Copper (Cu), Iron (Fe), Zinc (Zn) and Selenium significant (Snedecor and Cochran, 1980).
(Se) in testicular tissues were measured according
to the procedure which reported in AOAC (2004), 3. RESULTS
by using atomic absorption spectrophotometer The variations in the testes and epididymus
(Thermo Jarel Ash-AA-ScanI). weights of animals subjected to profenofos
treatment are shown in Table (1). There was
Data analysis and evaluation of statistical significant decrease (p<0.05) and (P<0.001) in
significance among different values determined weights of the testes and epididymus, respectively,
was done using the student's t-test. Statistical as compared to control group.

Table 1
Effect of oral administration of profenofos on testes and
epididymus weights of rats after sub-chronic exposure (60 days)
Profenfos-treated group 23.14
Parameter Control group
mg/kg body weight

Testes weight (g) 1.52 1.40

± 0.040 ± 0.004*

Epididymus weight (g) 0.37 0.02

± 0.014 ± 0.008***

Data represent mean ± SE, n = 5, * P< 0.05, *** P< 0.001 (Student’s t-test)

The effect of oral administration of significantly (p<0.05) in profenofos-treated group

profenofos for 60 days on sperm count and in comparison with the control group. Similarly,
motility in cauda epididymus is shown in Table (2). spermatozoal motility was also found to be
The spermatozoal density (count) increased significantly decreased (p<0.001).

Table 2
Effect of oral administration of profenofos on semen parameters in cauda
epididymus of rats after sub-chronic exposure (60 days):

Data represent mean ± SE, n = 5, * P<0.05 , *** P<0.001 (student’s t-test)

Results of testicular biochemistry have been (p<0.001, p<0.05 and p<0.01, respectively) in
depicted in Table (3). Alkaline (ALP), acid (ACP) profenofos-treated group as compared to control
phosphatese and lactate dehydrogenas (LDH) group. In addition, total protein level was found to
activities were recorded to have decreased

21
Influence of Subchronic Exposure of Profenofos on Biochemical Markers and Microelements in Testicular Tissue of Rats
Afaf A. El-Kashoury
____________________________________________________________________________________________________________

be significantly raised (p<0.05) in treated group in comparison with the control group.

Table 3
Effect of oral administration of profenofos on some testicular
biochemical parameters in rats after sub-chronic exposure (60 days)

Data represent mean ± SE, n = 4, * P<0.05,** P<0.01, *** P<0.001 (student’s test)

In addition to the findings listed above, series of spermatogenesis and high spermatozoal
we have observed the presence of microscopic concentration in the lumen (fig.1) Profenofos-
changes in the testes of male albino rats. intoxicated animals indicated that there were few
Histological findings of testes from control and numbers of sperm cells in the lumen of the
treated groups are presented in figs. 1, 2, seminiferous tubules (fig. 2), in correlation with
respectively. Normal control animals, revealed the control one.
normal mature seminiferous tubules with complete

Fig 1: Testes of rat in control has shown the normal

histological structure of the seminiferous tubules in nature active condition.

22
Marsland Press
Journal of American 2009: 5(1), 19-28
______________________________________________________________________________________

Fig 2: Testes of rat treated by profenofos showing low amount

of sperms in the lumen of the semineferous tubules.

Table 4
The Testicular tissue contents of microelements in
profenofos-treated rats after sub-chronic exposure (60 days)

Data presented mean ± SE of five individual values.

The effect of oral administration of levels of total protein and microelements (Cu, Fe,
profenofos for the 60 days on testicular tissue Zn and Se) in testicular tissues. Our results showed
contents of microelements is depicted in table that the weights of testes and epididymus were
(4).profenofos treatment produced significant significantly lower in the profenofos-treated rats
increase (p<0.001) in iron (Fe), copper (Cu), zinc than in the controls. The decrease in testicular
(Zn) as well as in selenium (Se) levels. weight in treated rats may be due to reduced tubule
size, spermatogenic arrest and inhibition of steroid
4. DISCUSSION biosynthesis of leydig cells, a site of steroid
Organophosphates (OPIs) are among the most biosynthesis (Sujatha et al. ,2001 and Kaur and
widely used synthetic insect pesticides. The wide mangat, 1980). The decrease in testicular weight in
spread use of OPIs has stimulated research into the profenofos-treated rats may indicate impairment at
possible extence of effects related with their testicular, pituitary, or hypothalamic level (Chitra
reproductive toxic activity (Joshi et al., 2007). The et al. ,1991). Similar results were recorded by Ref
present study results demonstrated that 60 day’s Joshi et al. (2007), who mentioned that
exposure of male rats to profenofos at the dose chlorpyrifos (OPIs) at dose levels of 7.5, 12.5 and
23.14 mg/kg body weight (4 doses/week) resulted 17.5 mg/kg b.wt./day, for 30 days, decreased
in decreased the testes and epididymus weights, significantly the weight of testes. The epididymus
male fertility indices (sperm count and motility), is androgen-dependant organ, relying on
and activities of ALP, ACP and LDH but increased testosterone for its growth and function

23
Influence of Subchronic Exposure of Profenofos on Biochemical Markers and Microelements in Testicular Tissue of Rats
Afaf A. El-Kashoury
____________________________________________________________________________________________________________

(Klinefelter and Hess, 1998). On discussing the ,1990) and thyroxine (T4) replacement in men
results with previous reports, it is proposed that with hypothyroidism is reported to improve sperm
profenofos probably imped the activity of testes motility (Kumar et al. 1990). Moreover, it had
and epididymus by inhibition of androgen been reported that chlorpyrifos brought about
production or its direct action on these organs marked reduction in epididymal and testicular
(Kaur and mangat, 1980), thus, the reduction in the sperm counts in exposed males (Joshi et al., 2007).
weights of testes and epididymus in our study may Also, testicular atrophy and degenerative changes
be due to lower bioavailability of androgen in the seminiferous tubules had been reported in
(Sujatha et al. ,2001). Moreover, the deleterious experimental animals administered with various
effects of profenofos on reproductive organ O’Ch and OPIs pesticides (Dutta and Dikshith,
weights might be due to a decrease in the 1973). Based on the data obtained in this study,
testosterone (T) and thyroid hormone levels after administration of profenofos into male albino rats
60 days from the onset of the treatment (Takizawa reduced the activities of acid and alkaline
and Horii, 2002). phosphatase and lactate dehydrogenase which
reflect suppression in testicular function (Johnson
The present results confirm the previous et al. ,1970). Activities of markers enzymes viz
reports of (El-kashoury and El-far, 2004) who ALP, ACP and LDH are considered to be
mentioned that administration of rats with functional indicators of spermatogenesis.
profenofos at 23.14 and 46.30 mg/kg body weight
for 28 days and 60 days, respectively, induced Our results confirm the findings of
significant decrease in thyroid hormone levels, (Salem et al. ,1989) who investigated the influence
there is ample evidence that thyroid hormone is of methamidophos (O’ps) on mammals. Results
essential to the normal development of testes in the showed that treatment of male rats with
neonate (Cook et al. ,1994 and Hardy et al. ,1996), methamidophos, at 100 ppm in drinking water for
as well as an elevation in cholesterol level, a 9 and 45 days, reduced significantly acid and
precursor of steroid hormone had occurred. alkaline phosphatase and lactate dehyrogenase in
Authors also, mentioned that inhibition of hepatic testicular tissue. Also, (Mustafa et al. ,2007)
microsomal 7-hydroxylation of cholesterol by reported that profenofos considered as one of the
profenofos leads to reduction of cholesterol break male reproductive toxicants. ALP is primary of
down and its accumulation. Sperm count is one of testicular and epididymual origin and, therefore,
the most sensitive tests for spermatogenesis and it suitable for differentiation of oligo-and
is highly correlated with fertility. Our results azoospermia (Turner and Sertich, 2001; Turner
revealed that, treatment of rats with profenofos and McDonell, 2003). Decline in ALP activity
significantly reduced the sperm count and motility. indicated that profenofos treatment produced a
The decreased sperm motility and density (count) state of decreased steroidogenesis where the inter
after oral administration of profenofos is may be and intercellular transport was reduced as the
due to androgen insufficiency (Chaudhary and metabolic reactions to channelize the necessary
Joshi, 2003) which caused impairment in testicular inputs for steroidogenesis slowed down
functions by altering the activities of the enzymes (Latchoumycandane et al. ,1997). Acid
responsible for spermatogenesis (Sinha et al. ,1995 phosphatases are enzymes capable of hydrolyzing
and Reuber, 1981). orthophosphoric acid esters in an acid medium.
Histological structure of the testes The testicular acid phosphatase gene is up-
confirmed the aforementioned results, where it is regulated by androgens and is down-regulated by
revealed degeneration in some of seminiferous estrogens (Yousef et al. ,2001). Activities of
tubules associated with low luminal spermatozoal phosphatases enzymes have been shown to rise
concentration. It is tempting to speculate that the when testicular steroidogenesis is increased
decreased sperm motility in the present study may (Mathur and Chattopandhyay, 1982).
have been related to our earlier studies on
profenofos (El-kashoury and El-far, 2004) which Also, (Latchoumycandane et al. ,1997)
pointed that subclinical hypothyroid state in rats mentioned that a decrease in ACP activity in free
administered with profenofos for 60 days had state would thus reflect decreased testicular
occurred. Also, men with hypothyroid have been steroidogenesis in rats and this may be correlated
reported to have lower sperm motility than with the reduced secretion of gonadotrophins.
euthyroid controls (Corrales – Hernandez et al. LDH is associated with the maturation of germinal

24
Marsland Press
Journal of American 2009: 5(1), 19-28
______________________________________________________________________________________
epithelial layer of seminiferous tubules and in the testicular tissues. On the other hand, similar
associated with post meiotic spermatogenic cells results were recorded by Al-Bayati et al. (1988),
(Sinha et al., 1997). An inhibition in the activity of who mentioned that 2, 3, 7, 8-tetrachlorodibenzo-
LDH in testes of profenofos-treated rats points p-dioxin (TCDD), O'ch, produced atrophy,
toward the interference of profenofos with the morphological changes and impaired
energy metabolism in testicular tissues spermatogenesis in testes of experimental animals.
(Mollenhauer et al., 1990). The correlation In addition, testicular tissue contents of Fe, Cu,
between LDH and motility and living sperm could and Zn were significantly increased in the treated
be a sign that extracellular LDH ensures rats. Zinc (Zn) markedly increased the ALP and
metabolism of spermatozoa, perhaps even in ACP activities and this occurred concomitantly
anaerobic conditions (Pesch et al., 2006). with the appearance of spermatids and mature
sperm cells (Guha and Vanha-Perttula, 1983).
As regards the testicular protein, results Selenium is an essential trace nutrient for humans
of the present study exhibit an increase in its level and animals. It is an essential at lower
in profefos-treated rats. The testicular fluid concentrations and toxic at higher concentrations.
contains both stimulatory factors as well as Se is required for normal testicular development
inhibitory factors that selectivity alter the protein and spermatogenesis in rats (Behne et al., 1996).
secretions (Brooks, 1983). Thus, the changes in The selinodeiodinase enzymes (types I, II and III
protein suggested that there is a reduction in the iodothyronine deiodinase) control the metabolism
synthetic activity in testes. An elevation in of thyroid hormone, which is essential for the
testicular protein in the present study confirms the normal development (Defrance et al., 1995) and
previous results by (Joshi et al., 2007) who function (Latchoumycandane et al., 1997) of testes
mentioned that the protein content was raised at in rats. The above explanation supports our
significant levels in chlorphrifos-treated rats. findings where elevated testicular tissue content of
Gupta et al. (1981) and Singh and Pandey (1989) Se associated with decrease in testicular weight,
illustrated that an elevation in the testicular protein sperm count and motility in profenofos-treated
may be due to the hepatic detoxification activities rats. In support of these findings, earlier results
caused by endosulfan (O'ch) which results in the (El-Kashoury and El-Far, 2004) revealed that
inhibitory effect on the activities of enzyme treatment of rats with profenofos at the same dose
involved in the androgen biotransformation and time interval decreased markedly (T3) level in
(Dikshith and Dutta, 1972). plasma in comparison with the control group.

Similar results showed the same trend in Cupper is necessary for many enzymes
the protein content caused by several pesticides, at like the Cu-Zn-Superoxide dismutase (SOD),
different periods and / or different concentrations, which is involved in cell protection against free
had been also reported (Shivanandappa and (Oxygen) radicals. Copper is also needed for the
Krishnakumari (1981), Bhatnagar and Malviya, cytochrome C oxidase that is responsible for
1986; Chitra et al., 1999; Choudhary and Joshi, energy supply and for cellular and humoral
2003). In accordance with the findings of the immunity (Leonhard-Marek, 2001). As regards Cu
present study, Rao and Chinoy (1983), suggested concentrations, an administration of rats with
that the accumulation of protein occurred in testes profenofos increased testicular tissue contents of
and epididymus due to androgen deprivation to Cu by 2-fold, respectively. Elevated Cu
target organs. This deprivation effect also led to a concentrations reduced oxidative processes and
reduction in testicular and cauda epididymus glucolysis that may cause immotility and reduced
sperm population, loss of motility in the latter and viability (Leonhard-Marek, 2001). A proposed
an increase in number of abnormal spermatozoa, mechanism could explain elevated iron
thereby manifesting 100% failure in treated concentrations in testicular tissues in profenfos-
animals. Results of the present investigation treated rats, is that iron is known to be essential
showed that administration of profenofos into male and mostly bound to transferrin (produced by
rats increased the concentration of trace elements; sertoli cells), haptoglobin (sertoli, leydig and germ
Cu, Fe, Zn and Se in testicular tissue, which have a cells) and lactoferrin (spermatozoa and vascular
pivotal role in spermatogenesis (Homma-Takeda et gland). These proteins contain catalytic inactive
al., 2007). These findings are not in accordance iron which avoids extensive oxidation (Leonhard-
with those of Salem et al. (1989), who stated that Marek, 2001). Results of the present investigation
treatment of rats with methamidophos (OPIs), for suggested that profenofos may impede the
45 days, decreased the concentrations of Zn and Se utilization of micro-elements in the testes,

25
Influence of Subchronic Exposure of Profenofos on Biochemical Markers and Microelements in Testicular Tissue of Rats
Afaf A. El-Kashoury
____________________________________________________________________________________________________________

consequently stagnation of Cu, Fe, Zn and Se in two dimensional gel electrophoresis. Mol.
the testes occurred. It is concluded that profenofos Cell Endocrinol., 29 : 255-270.
induced adverse effects on testicular function by 10. Chantcli-Forti, G.; Paolini, M. and Hrelia,P.
altering biomarker enzymes activities as well as (1993). Multiple end point procedure to
disrupting micro-elements levels, thus care should evaluate risk from pesticides. Environ. Health
be taken and more studies should be done to perspect. 101, 15-20.
increase the validity of those information. 11. Chaudhuri,K.; Selvaraj, S. and Pal, A. K.
( 1999). Studies on the genotoxicity of
endosulfan in bacterial systems. Mutat. Res.
Abbreviation used: 439, 63-67.
OPIs, organophosphorous insecticides; 12. Chia, S.E.; Ong, C.N.; Chua, L.H.; Ho, L.M.
O'Ch, organochlorine, TP, total protein, ALP, and Tay, S.K. (2000). Comparison of zinc
alkaline phosphatase; ACP, acid phosphatase; concentrations in blood and seminal plasma
LDH, lactate dehydrogenase; Cu, Copper; Zn, and the various sperm parameters between
Zinc; Fe, Iron; Se, Selenium; Ec, Emulsifiable fertile and infertile men, J Androl. 21: 53-57.
concentrate; T4, Thyroxine; T3, Triiodothyronine; 13. Chitra, K.C.; Latchoumycandane, C. and
T, Testosterone; Ros, Reactive oxygen species. Mathur, P.P. (1999). Chronic effect of
endosulfan on the testicular functions of rat.
REFERENCES Asian J. Andrology, 1 : 203-206.
1. A.O.A.C. (2004). Association of official 14. Choudhary, N. and Joshi, S.C. (2003).
analytical chemistry. Reproductive toxicity of endosulfan in male
2. Al-Bayati, Z.A.; Wahab, Z.Z. and Stohs, S.J. albino rats. Bull. Environ. Contam. Toxicol.,
(1988). 2, 3, 7, 8 Tetrachlorodibenzo-p-dioxin 70 : 285-289.
(TCDD)-induced alterations in lipid 15. Cooke, P.S.; Zhao, Y.D. and Bunick, D.
peroxidation, enzymes and divalent cations in (1994). Triiodothyronine inhibits proliferation
rat testis. Xenobiotica. 18: 1281-1289. and stimulates differentiation of cultured
3. Babson, A.L. and Ready, A.P. (1959). neonatal sertoli cell: Possible mechanism for
Colourimetric method for the determination of increase adult testis weight and sperm
total and prostatic acid phosphatase. Am. J. production induced by neonatal goitrogen
Clin. Path., 32 : 89-91. treatment. Biol. Reprod. 51: 1000-1005.
4. Babson, L.A. (1965). Phenolphalein 16. Corrales-Hernandez, J.J.; Miralles-Gareia, J.M.
monophosphate methods for the determination and Garcia-Diez, L.C. (1990). Primary
of alkaline phosphatase. Clin. Chem., 11 : 789. hypothyroidism and human spermatogenesis.
5. Banchraft, J.D.; Stevens, A. and Turner, D.R. Arch. Androl. 25: 21-27.
(1996). Theory and practice of histological 17. Defrance, L.R.; Hess, R.A.; Cooke, P.S. and
techniques. 4th Ed., Churchill Livingstone, Russell, L.D. (1995). Neonatal
New York, London, San Francisco, Tokyo. hypothyroidism causes delayed sertoli cell
6. Behne, D.; Weiler, H. and Kyriakopoulos A. maturation in rats treated with
(1996). Effects of selenium deficiency on propylthiouracin: Evidence that sertoli cell
testicular morphology and function in rats. J. controls testis growth. Anat. Rec. 242: 57-69.
Repord Fertile. 106: 291-297. 18. Dikshith, T.S.S. and Dutta, K.K. (1972).
7. Bhatnagar, V.K. and Malviya, A.N. (1986). Pathological changes induced by pesticides in
Changes in some biochemical indices in rat the testes and liver of rats. Exp. Pathol., 7 :
upon pesticide toxicity. Indian J. Biochem. 309-316.
Biophys., 15 : 78-81. 19. Dutta, K.K. and Dikshith, T.S.S. (1973).
8. Bradford, M.M. (1976). A rapid and sensitive Histopathologic changes in the testes and liver
method for the quantitation of microgram of rats repeatedly exposed to pesticides. Exp.
quantities of protein utilizing the principle of Pathol., 8 : 363-370.
protein-dye binding. Analyt. Biochem., 72 : 20. El-Kashoury, A.A. and El-Far, F.A. (2004).
248-254. Effect of two products of profenofos on
9. Brooks, D.E. (1983). Effect of androgen on thyroid gland, lipid profile and plasma APO-
protein synthesis and secretion in various 1/FAS in adult male albino rats, Egypt. J.
regions of the rat epididymis, as analysed by Basic and Appl. Physiol., 3: 213-226.

26
Marsland Press
Journal of American 2009: 5(1), 19-28
______________________________________________________________________________________
21. Feustan, M.H.; Bodnai, K.R. and Kerstetter, accessory sex glands. In : Reprod. and Dev.
S.L. (1989). Reproductive toxicity of 2- Toxicol., pp. 553-591, New York, Basel,
methoxy ethanol applied dermally to occluded Hong Kong; Marcel Dekker, Inc.
and non-occluded sides in male rats. Toxicol. 34. Kumar, B.J.; Khurana, M.L.; Ammini, A.C.;
Appl. Pharmacol., 100 : 145-165. Karmarkar, M.G. and Ahuja, M.M.S. (1990).
22. Gangadharan, B.; Murugan, M.A. and Mathur, Reproductive endocrine functions in men with
P.P. (2001). Effect of methoxychlor on primary hypothyioidism: Effect of thyroxine
antioxidant system of goat epididymal sperm replacement. Horm. Res. 34: 215-218.
in vitro. Asian J. Androl., 3 : 285-288. 35. Latchoumycandane, C.; Gupta, S.K. and
23. Guha, K. and Vanha-Perttula, T. (1983). Acid Mathur, P.P. (1997). Inhibitory effects of
phosphatases in the mouse testis: activity hypothyrodism on the testicular functions of
changes during development. Arch-Anorl. 10 postnatal rats. Biomed. Lett., 56 : 171-177.
(1): 7-16. 36. Leonhard-Marek, S. (2001). Influence of
24. Gupta, P.K.; Shrivastava, S.C. and Ansari, drugs, pollution and trace elements on male
R.A. (1981). Toxic effects of endosulfan on fertility. In: Busch, W. and Holzmann, A.
male reproductive organs in rats. Indian J. Editors, Andrology in veterinary medicine,
Biochem. Biophys., 18 : 159-163. Stuttgart, Schattauer, pp. 474-481.
25. Hamameh, S. and Gatti, J.L. (1992). Role of 37. Mathur, P.P. and Chattopadhyay, S. (1982).
the ionic environment and internal pH on Involvement of lysosomal enzymes in
sperm activity, Hum Repord Suppl. 4: 20-30. flutamide-induced stimulation of rat testis.
26. Hardy, M.P.; Sharma, R.S.; Arambepola, N.K.; Andrologia, 14 : 171-176.
Sottas, C.M.; Russell, L.D.; Bunick, D.; Hess, 38. Mollenhauer, H.H.; Morre, D.J. and Rowe,
R.A. and Cooke, P.S. (1996). Increased L.D. (1990). Alteration of intracellular traffic
proliferation of leydig cells induced by by monensin : mechanism, specificity and
neonatal hypothyroidism in the rat. J. Androl. relationship to toxicity. Biochem. Biophys.
17: 231-238. Acta, 1031 : 225-246.
27. Homma-Takeda, S.; Nishimura, Y.; Watanabe, 39. Moss, D. W. and Handerson, A.R. (1994).
Y. and Yukawa, M. (2007). Site-specific Lactate dehydrogenase 2nd Ed., Textbook of
changes in zinc levels in the epididymus of clinical chemistry, chapter 20, Enzymes, pp.
rats exposed to ionizing radiation. Nuclear 812-818
instruments and methods in physics research 40. Moustafa, G.G.; Ibrahim, Z.S.; Hashimoto Y;
section B. Beam interactions with materials Alkelch A.M.; Sakamoto K.Q.; Ishizuka M.
and atoms. 260: 236-239. and Fujita S. (2007). Testicular toxicity of
28. Huang, Y.L.; Tsen, W.C.; Cheng, S.Y. and profenofos in matured male rats. Arch Toxicol.
Lin, T.H. (2000). Trace elements and lipid 41. Pandy, V.K.; Parmeshwaran, M. and Soman,
peroxydation in human seminal plasma, Biol. S.D. (1983). Concentrations of
Trace element. 76: 207-215. morphologically normal, motile spermatozoa:
29. Jadaramkunti, U.C. and Kaliwal, B.B. (2002). Mg, Ca and Zn in the semen of infertile men,
Dicofol formulation induced toxicity on tests Sci Total Environ 27: 49-52.
and accessory reproductive organs in albino 42. Pesch, S.; Bergmann, M. and Bostedt, H.
rats. Bull. Environ. Contam. Toxicol., 69 : (2006). Determination of some enzymes and
741-748. macro-and microelements in stallion seminal
30. Johnson, A.D.; Gomes, M. and Vandemark, plasma and their correlations to semen quality.
N.L. (1970). The testis. 1st Ed., Academic Theriogenology. 66: 307-313.
Press, New York and London. 43. Rao, M.V. and Chinoy, N.J. (1983). Effect of
31. Joshi. S.C.; Mathur, R.; Gilati and Tags, N. estradiol benzoate on reproductive organs and
(2007). Testicular toxcity of chlorpyrifos (an fertility in the male rat. Eur. J. Obstet.
organophosphate pesticide) in albino rat. Gynecol. Reprod. Biol., 15 (3) : 189-198.
Toxicol Ind Health. 23: 439-444. 44. Reuber, M.D. (1981). The role of toxicity in
32. Kaur, C. and Mangat, H.K. (1980). Effects of the carcinogenicity of endosulfan. Sci. Total.
estradiol dipropionate on the biochemical Environ., 20 : 23-47.
composition of testes and accessory sex 45. Salem, H.A.; Younis, M.; El-Tohamy, M.; El-
organs of adult rats. Andrologia, 12 (4) : 373- Herrawie, M.; Soliman, M. and Khalaf, A.
378. (1989). Thyrotrophic and thyroid hormones
33. Klinefelter, G.R. and Hess, R.A. (1998). concentration and some testicular biochemical
Toxicology of the male excurrent ducts and parameters as affected by oral administration

27
Influence of Subchronic Exposure of Profenofos on Biochemical Markers and Microelements in Testicular Tissue of Rats
Afaf A. El-Kashoury
____________________________________________________________________________________________________________

of methamidophs (Organophosphorus) into fertile and infertile men, Fertile Steril. 40:
adult male rats. J. Egypt. Vet. Med. Ass. 49: 670-677.
667-675. 53. Sujatha, R.; Chitra, K.C.; Latchoumycandane,
46. Sharma, Y.; Bashir, S.; Irshad, M.; Nag, T.C. C. and Mathur, P.P. (2001). Effect of lindane
and Dogra, T.D. (2005). Dimethoate-induced on testicular antioxidant system and
effects on antioxidant status of liver and brain steroidogenic enzymes in adult rats. Asian J.
of rats following subchronic exposure. Androl., 3 : 135-138.
Toxicology, 215 : 173-181. 54. Sultatos, L.G.(1994). Mammalian toxicity of
47. Shivanandappa, T. and Krishnakumari, M.K. organophosphorus pestiside. J. Toxicol.
(1981). Histochemical and biochemical Environ. Health 43, 271-289.
changes in rats fed dietary Benzene 55. Takizawa, S. and Horii, I. (2002).
hexachloride. Indian J. Exp., 19 : 1163-1168. Endocrinological assessment of toxic effects
48. Singh, S.K. and Pandey, R.S. (1989). on the male reproductive system in rats treated
Differential effects of chronic endosulfan with 5-fluorouracil for 2 or 4 weeks. J.
exposure to male rats in relation to hepatic Toxicol. Sci., 27 (1) : 49-56.
drug metabolism and androgen 56. Turner, R.M. and McDonnell, S.M. (2003).
biotransformation. Indian J. Biochem. Alkaline phosphatase in stallion semen :
Biophys., 26 : 262-267. characterization and clinical applications.
49. Sinha, N.; Narayan, R. and Saxena, D.K. Theriogenology, 60 : 1-10.
(1997). Effect of endosulfan on the testes of 57. Turner, R.M. and Sertich, P.L. (2001). Use of
growing rats. Bull. Environ. Contam. Toxicol., alkaline phosphatase activity as a diagnostic
58 : 79-86. tool in stallions with a zoospermia and
50. Sinha, N.; Narayan, R.; Shanker, R. and oligospermia. Anim. Reprod. Sci., 68 : 315-
Saxena, D.K. (1995). Endosulfan-induced 316.
biochemical changes in the testis of rats. Vet. 58. Weil, C.S. (1952). Tables for convenient
Hum. Toxicol., 37 (6) : 547-549. calculation of medium effective (LD50 or
51. Snedecor, G.W. and Cochran, W.G. (1980). EC50) and instruction in their use. Biometrics,
Statistical methods (7th ed). Lowa State 8 : 249-263.
University Press. Ames. 59. Yousef, G.M.; Diamandis, M.; Jung, K. and
52. Stanwell-Smith, R.; Thompson, S.G.; Haines, Eleftherios, P. (2001). Molecular cloning of a
A.P.; Ward, R.J. Cashmore, G. and novel human acid phoshatase gene that is
Stedronska, J. (1983). A comparative study of highly expressed in the testes. Genomics, 74
zinc, copper, cadmium and lead levels in (3) : 385-395.

28
Marsland Press
Journal of American Science 2009: 5(1), 29-34
______________________________________________________________________________________
Research Article
Effect of Duckweed Meal on The Rate of Mold Infestation In Stored
Pelleted Fish Feed
1
B.N. Effiong, 2A. Sanni,
1
Dept of Fisheries Technology, Federal College of Freshwater Fisheries Technology
New Bussa, Nigeria.
2
Dept of Microbiology, University of Ilorin, Nigeria.
________________________________________________________________________________
ABSTRACT: The effect of duckweed (Lemna pauciscostata) meal on the rate of mould infestation
in stored pelleted fish feed was carried out. Freshly harvested duckweed was dried and thoroughly
ground into powder using a milling machine. Five dry fish feeds were then prepared using duckweed
as a replacement for fishmeal at 0%, 10%, 20% and 30% respectively at 40% crude protein, a diet
for catfishes. The resultant pelleted feeds were sun dried for 24hrs and stored in airtight polyethylene
bags at room temperature. Quantitative mold count using direct colony counts on pour plate
technique with 24hr old culture was carried out bi-weekly until profuse growth were recorded within
24hrs in all experimental feeds. Results showed that mold count from experimental feeds decreased
with increasing concentration of duckweed. Ethanolic extracts also showed higher inhibitory
properties on radial mycelial growth of all the isolates. Isolates identified were Fusarium oxysporium,
Penicillium digitatum, Aspergillus niger, A.fumigatus, A..flavus, Rhizopus stolonifer and R..oryzae.
[Journal of American Science 2009: 5(1), 29-34] (ISSN: 1545-1003)

Key words: Polyethylene, Fusarium oxysporium

1. INTRODUCTION despite the enormous harmful effect it poses on the

Feeds are a major cost input into the aquaculture development of aquaculture in Nigeria (Effiong
industry and their insufficiency is prominent and Eyo, 2001). Duckweed meal has been reported
among the factors responsible for inadequate to resist attacks by mould for more than 5 years
aquacultural production of fish. (Skillicorn et al., 1993). Duckweed meal is the
Compounded feeds are prepared with biologically compounded form of the group of aquatic
decomposable materials. These materials macrophytes from the family Lemnaceae. The
decompose while in storage due to environmental dried powdered and directly pelleted forms of this
factors such as temperature and humidity. Change plant have been observed in storage for 13 years
in temperature and humidity affects the moisture without any signs of fungal growth or physical
content of compounded feed as well as the rate at damage, retaining its nutrient content (Mbagwu,
which chemical changes takes place thereby 2001). This study is therefore aimed at
enhancing invasion and growth of fungi in the feed determining the effect of duckweed meal on the
(Sena and Anderson, 1995; Effiong and Eyo, rate of mould infestation in stored pelleted fish
1999). Recontamination of feedstuffs by feeds.
adventitious microorganisms during storage is of
primary concern to the feed processor. 2. MATERIALS AND METHODS
Freshly harvested duckweed was thoroughly rinsed
Moulds are the principal spoilers of with clean water and evenly spread on a mosquito
feedstuff in storage (Chow, 1980). Moulds net-size mesh outside to sundry and thereafter
infestation reduces the nutritional value of feeds dried in a forced air oven at 165 0c for 48 hours
through loss of dietary lipids and amino acids and ground to powder with a milling machine
(Jones, 1987). They also produce mycotoxins, according to Mbagwu and Adeniji (1987).Five dry
which cause staleness of feed. He also stated that diets were prepared in which fish meal was
there is no effective way of eliminating fungal replaced with duckweed at 0%, 10%, 20% and
growth in stored pelleted feed. Their growth can 30% levels using the method of Akegbejo Samson
only be controlled. Research work on the problems (1999) at 40% crude protein, a diet for catfishes.
of storage of feedstuff \feed has been rather scanty The various feed ingredients were thoroughly

29
Effect of Duckweed meal on the rate of mold infestation in stored pelleted fish feed.
B.N. Effiong et al.
____________________________________________________________________________________________________________

ground into fine meal and mixed together with 3. RESULTS AND DISCUSSION.
vitamin premix and salt using hot water. The bi-weekly fungal count (cfu) from
the experimental feed at varying concentrations of
The resultant mixture was pelleted with duckweed showed decrease in fungal growth with
Moulinese HV6 model pelleting machine and sun increasing concentration of duckweed (Table 1).
dried for 24 hours. The diets were stored in airtight This observation could be attributed to the
containers at room temperature for 2 weeks. 1.0g antifungal properties of duckweed acting against
of each feed sample were ground using pestle and the growth of fungal species in the feed. Skillicorn
mortar, to prepare 10-fold serial dilution. Agar was et al., (1993) attributed the long storage
prepared using sterilized glasswares according to characteristic of duckweed meal to the presence of
manufacturer’s instruction and autoclaved at 121 high levels of wax. It could be possible that wax
0
c for 15 minutes. It was allowed to cool to about presents physical barriers to the growth of molds,
37 0c before 1% streptomycin was added to which might impair their utilization of nutrients in
prevent bacterial contamination (Nwachukwu, the feeds. The molds isolates from the
1988). A 48hour old culture of the isolates were experimental feed samples were Fusarium
subcultured and incubated at room temperature to oxysporium, Penicillium digitatum. Aspergillus
produce pure cultures from which stock were niger, A. fumigatus, A. flavus, Rhizopus stolonifer
prepared and stored. A bi-weekly mould count and R. oryzae. Chow (1980) reported that the most
from each experimental diet was carried out common molds involved in the spoilage of
quantitatively using direct colony count on pour feedstuffs belong to the Aspergillus and
plate technique (Miles and Misra, 1938) with 24- Penicillium species among others. The presence of
hour-old culture. Enumeration continued until Aspergillus flavus from the feed indicates the
profuse growth was recorded within 24 hours in all possibility of mycotoxins, compounds produced by
the experimental diets. Mould isolates were this species that are toxic to both humans and fish.
characterized during sporulation on the basis of Feedstuffs known to be contaminated by A. flavus
cultural and morphological characteristics as well include groundnut cake, maize, sorghum,
as microscopic examination (Samson and Reenen- cottonseed cake, copra and cassava (Chow, 1980).
Hoekstra, 1988). Sample of duckweed meal was The same author however reported that for
ground using an Automatic Weed Grinder after it aflatoxins to be produced, A. flavus must be
was thoroughly washed and air-dried. 5g of this present alone in a practically pure culture and that
each was measured and blended with 25ml of the presence of other molds, yeast or even bacteria
sterile distilled water (Oyagade, 1994). After seems to interfere with aflatoxin production. These
thoroughly blending for 7 minutes, the slurry was findings have also been reported by Abdulhamid
filtered through a four –layered muslin cloth. The (2008).
filtrate was passed through a 0.48 millimicron
The effect of duckweed extracts on the
Millipore filter and transferred into sterile bottle.
radial mycelial growth of fungal isolates from the
In order to compare the efficiency of the extraction
experimental feeds is shown in Table
process, 95 % alcohol was used as the comparative
2.Differential efficacy on the test organisms was
solvent using the same method.
observed between the aqueous and ethanolic
Radial mycelial growth inhibition tests
extracts of duckweed meal. Ethanol appeared
were carried out on the isolates (Van-Etten, 1973;
better as an extractant judging from the wider
Oloke et al., 1988). The extracts were separately
activity spectrum and the resultant effect on the
incorporated into molten PDA at 18ml of media to
isolates. This observation perhaps suggests the
2ml of extract. Control plates had either sterile
possibility of the occurrence of bioactive
water or ethanol without extract.
substances that are not only soluble in water but
Agar- extract mixtures were poured into sterile
also in organic solvent in the plant material.
glass petri dishes and allowed to set (Adedayo,
Majekodunmi et al., (1996), and Martinez et al.,
1994). Mycelial plugs of the test organisms of
(1996) reported that a higher activity of extractable
5.0mm diameter were cut using sterile cork-borer
natural products were obtained in ethanol
from the advancing margin of the fungal colonies.
compared with aqueous extracts. Odemena and
These were placed at the center of PDA containing
Essien (1995) also reported that the bacterial
concentrations of 5% sterile distilled water or
activity of alcoholic extracts of the roots of fluted
ethanol. All plates were incubated at 25 0c and
pumpkin, Telfaria occidentalis was better than that
radial mycelial growth recorded for 72 hours at 24
of aqueous extracts. Natarajan et al., (2005)
h0urs interval
reported the antifungal properties of three

30
Marsland Press
Journal of American Science 2009: 5(1), 29-34
______________________________________________________________________________________
medicinal plant extracts against Cercospora relative humidity remained constant between 79
arachidicola. They reported that fungal growth was and 80%. According to Chow (1980), growth of
gradually suppressed with increasing extract fungi is only possible at temperature above 250c
concentration. Similar findings have been reported and relative humidity values at 65%. There any
by Lucia et al., (2002), Silva et al., (2001) and reduction in fungal growth in the experimental
Costa et al., (2000). These reports are similar to feeds could not be attributed to directly affect the
the findings of this study.Olafimihan (2003) rate of fungal infestation of compounded feed in
working on the antibacterial properties of aqueous storage.
and ethanolic extracts of Neem plant reported that
the antibacterial activity of the concentrated 4. CONCLUSION
extract increased with increase in its concentration. The results obtained from this study indicate
This report is similar to the findings from this reduced growth performance in the fungal species
study with he observation that increasing isolated from the experimental feed which also
concentration of duckweed meal in experimental signified low infestation rate. Fungal growth
feed resulted in decreasing fungal growth. decreased generally with increase in concentration
of duckweed meal in feed samples.
The environmental conditions of
temperature and relative humidity during the
The result of this experiment have shown that
period of the study were high and fell within the
duckweed has the potential of being a beneficial
ranges that support luxuriant growth of molds in
agent for the control of fungal growth in
the experimental feed sample. The temperature
compounded feed in storage.
range varied between 27.2 and 30.6 0c while

Table 1
Percentage composition of experimental feed with different
inclusion levels of duckweed meal

Ingredients (g) 0% 10% 20% 30%

Duckweed 0 2.6 5.2 7.8

Fish meal 26 33.4 20.8 18.2

Yellow maize 48 48 48 48

Soya Bean meal 15 15 15 15

Groundnut cake 6 6 6 6

Vitamin premix 2 2 2 2

Bone meal 2.5 2.5 2.5 2.5

Salt 0.5 0.5 0.5 0.5

Total 100 100 100 100

31
Effect of Duckweed meal on the rate of mold infestation in stored pelleted fish feed.
B.N. Effiong et al.
____________________________________________________________________________________________________________

Table 2
Bi- weekly fungal counts at varying concentrations
of duckweed in experimental feed.

Concentration Fungal count (cfu\ml)(x 107)

of Time (wk)

Duckweed 2 4 6 8 10 12 14 16

(%).

0 12 20 31 43 72 Profuse Profuse Profuse

10 5 9 21 35 52 108 Profuse Profuse

20 3 11 18 27 48 90 Profuse Profuse

30 - - 9 16 21 54 76 Profuse

Table 3
Effect of duckweed extracts on the radial mycelial growth of fungal isolates

Test Mycelial growth (mm)

Organism Aqueous Extract Ethanolic Extract
0% 5% 10% 0% 5% 10%
Fusarium 46 21 10 10 - -
oxysporium
Penicillium 50 35 24 9 5 -
digitatum
Aspergillus 47 27 18 16 7 2
niger
Aspergillus 38 18 12 4 - -
fumigatus
Aspergillus 50 38 20 16 - -
flavus
Rhizopus 36 29 16 14 - -
oryzae
Rhizopus 42 21 13 22 10 4
stolonifer

32
Marsland Press
Journal of American Science 2009: 5(1), 29-34
______________________________________________________________________________________

Table 4
Proximate composition of experimental feed with different inclusion level of duckweed

Feed Sample % Crude %Ether %Ash %Moisture %Crude fibre

protein extract content content
0% 43.35 14.02 12.30 1.00 6.50

10% 42.56 14.29 12.00 1.00 4.46

20% 41.87 12.83 11.90 2.00 5.13

30% 45.06 11.76 13.29 2.00 4.90

REFERENCES Proceedings of the 12th Annual Conference of

1. Abdulhamid, A.M (2008). Mycotoxicoses in Biotechnology Society of Nigeria.Pp111-114.
fish with special emphasis on the Egyptian 8. Jones, F. (1987). Controlling mould growth in
Situation.E:\nicotoxins in feeds.htm. feeds. Feeds International.8: 20-29.
2. Akegbejo-Samson,Y (1999). Growth response 9. Lucia,K.H.S;Cecilia, A.Pedro, N.Suzana,C.S,
and nutrient digestibility by Clarias Judasio,G.DO; Andre,T.B.M; Luciano,M.L
gariepinus fed varying levels of dietary and Maria,R.R Silva (2002). Antifungal
periwinkles flesh as replacement for fishmeal properties of Brazilian Cerrado plant. Braz. J.
in low cost diets. Journal of Applied Microbiology. Vol.33 (3) 102-107.
Trop.Agriculture.Vol.4 (1) 37-41. 10. Mbagwu,I.G and Adeniji,H.A(1988).The
3. Adedayo, O and Kolawale, P.O (1994). nutritional content of duckweed (Lemna
Resistance of mouse-virulent encapsulated pauciscostata Hegelm) in the Kainji Lake area,
nasal isolates of Staphylococcus aureus to Nigeria. Aquatic Botany. (29) 357-366.
disinfectant and antiseptics. Biomedical 11. Mbagwu, I.G (2001). The effect of long-term
Letters. (50) 151-156. storage on the nutrient characteristics of
4. Chow, K.W (1980). Storage problems of Duckweed (Lemna pauciscostata Hegelm).
feedstuffs. Fish Feed J.Arid Agric. (11) 147-149.
Technology.ADCP/REFP\80\11,UNDP\FAO, 12. Miles, A.A and Misra, S.S (1938). Estimation
Rome 215-224. of bactericidal power of blood. J. Hyg (38)
5. Costa, T.R; Fernandes, O.F.L; Santos,S.C; 732-749.
Oliveira, C.MA; Liao,L.M; Ferri,P.H; 13. Nwachukwu, C.O (1988). Microbiology of
Paula,J.R.P; Ferreira,H.D; Sales,H.N and Pepper (C.annum) and the efficacy of some
Silva,M.R.R (2000). Antifungal activity of local methods of preservation. M.Sc Thesis.
volatile constituents of Eugenia dysentrica Biological Sciences.UniIlorin.
leaf oil. J. of Ethnopharmacol.72:11-117. 14. Natajaran,D; Srinivasan,K; Mohanasundari,C;
6. Effiong, B.N and Eyo, A.A (2001). Quality Perumal,G; Dheen, M.A.N;Ganapathi,G.A
deterioration of feeds and feedstuff in storage- and Rajarajan,T. (2005).Antifungal properties
A review. Fish nutrition and Fish Feed of three medicinal plant extracts against
Technology.ISBN 978-177-046-5 Pp113-121. cercospora arachidicola. Advances in plant
7. Effiong, B.N and Eyo,A.A (1999).Control of Sciences Vol.18 (1) 45-47.
mould infestation in stored pelleted feeds.

33
Effect of Duckweed meal on the rate of mold infestation in stored pelleted fish feed.
B.N. Effiong et al.
____________________________________________________________________________________________________________

15. Odemena,C.S and Essien ,J.P alcoholic extracts of the neem plant
(1995).Antibacterial activity of the root (Azadirachta indica. A.Juss) Ph.D Thesis.
extract of Telfera occidentalis (Fluted UniIlorin, Nigeria.
Pumpkin). West African Journal of Biological 19. Sena,S.S and Anderson, T.A (1995). Fish
and Applied Chemistry 40(1-4): 29-32. Nutrition in Aquaculture. Champman and Hall
16. Oloke, J.K; Kolawole,D.O and Erhun, w.o Aquaculture Series.1st Edition. Champman
(1988). The antimicrobial and antifungal and Hall, 2-6 Boundary Row,London
activities of certain components of SE118HN.
Aframonium meleguets fruits.Fitoterapia 20. Skillicorn, P; Spirar, W; and Journey, W
59(5)384-388. (1993). Duckweed Aquaculture. A New
17. Oyagade,J.O (1994) Antimicrobial efficacy of Aquatic Farming System for developing
stem bark extracts of two Nigerian medicinal countries. A World Bank Publication.
plants,Terminalia glaucescens (Planch) and National Agricultural Research Project
Entada africana (Guill and Perr.)Ph.D (NARP), Nigeria.309Pp.
Thesis,Biological Sciences, UniIlorin,Nigeria. 21. Van-Etten,H.D (1973). Differential Sensitivity
18. Olafimihan,C.A (2003).Studies on the of fungi to Pistat and Phaseolin.
antibacterial properties of aqueous and Phytochemistry (63) 1477-1482.

34
Marsland Press
Journal of American Science 2009: 5(1), 35-43
____________________________________________________________________________________________________________
Research Article
Distribution and Sources of Organochlorine Pesticides (OCPs) in Karst
Cave, Guilin, China
*
Annette Sylvie Muhayimana1, *Qi Shihua1, Wang Yinghui 1,
Kong Xiangsheng 1, Odhiambo Joshua Owago1, Zhang Junpeng1
(1) Department of Environmental Engineering, Faculty of Environmental Studies,
Key Laboratory of Bio and Environmental Geology of Ministry of Education, China University of
Geosciences, Wuhan, Hubei 430074, China
_____________________________________________________________________________________________________

Abstract: Despite the numerous researches done on Organochlorine pesticides (OCPs) in China and
in the world, information regarding emissions and concentrations of OCPs in Karst caves is
extremely limited. Karst areas have much higher ecological vulnerability and are so easy to be
contaminated. This paper presents results of a monitoring program conducted in Dayan cave, Guilin,
China that was designed to characterize levels, trends and sources of pesticides in soil (sediment)
samples. Thirteen soil samples were collected and OCPs were analysed. Inside the cave a total
concentration of OCPs (∑OCPs) detected was 29.659 ng /g with a mean value of 3.295 ng /g and
∑OCPs detected outside the cave was 74.108 ng /g with a mean value of 18.527ng/g. ∑OCPs
outside the cave was higher than ∑OCPs outside the cave. The concentration of Chlordane in OCPs
was highest among all the OCPs detected with range of 0.12─13.253ng/g and mean value of 3.93 ng
/g. The next compound with high level of concentration was Heptachlor which ranged from Non-
detected (ND) to 2.465 ng/g with a mean value of 1.4 ng/g. The pollution of OCPs in soil comparing
with other countries and other areas in China was light. The analysis of
Dichlorodiphenyltrichloroethane (DDT) and Hexachlorocyclohexane (HCH) isomers showed that
there was fresh input of Dicofol and Lindane in the study area. By calculating the ratios of
Dichlorodiphenyldichloroethane (DDD) to Dichlorodiphenyldichloroethylene (DDE), it was found
that the degradation of DDT outside the cave was aerobic and the degradation of DDT inside the
cave was anaerobic. [Journal of American Science. 2009;5(1):35-43]. (ISSN: 1545-1003).

Keywords: Organochlorine pesticides, Karst cave, Soil, Guilin, China.

1. INTRODUCTION greatest application and current residue levels was

found strong (Shivaramaiah et al., 2002). The
Organochlorine pesticides are a group of release of OCPs from soils continues to be a
persistent organic pollutants (POPs) which are to source of OCPs pollution to the environment
be eliminated or reduced on their release into the (Meijer et al., 2001).
environment in many countries. Because of their
persistence in the environment, and biological China is a large producer and consumer of
accumulation through the food web, OCPs can Pesticides in the world (Rongbing et al., 2006).
cause environmental damage, and affect human Large amount of OCPs were used in past decades
health (Colborn et al, 1996) .Due to their volatility to sustain over population in China. HCH and
and persistence in the air; OCPs are subjected to DDT were widely used in China from 1952-1983.
long-range atmospheric transport (LRAT). Although their use had been discontinued in China
Therefore, OCPs released in the tropical and since 1983, their persistence has left residual
subtropical environments could be dispersed amounts in the soil in many areas (Zhao Ling and
rapidly through air and water, and tend to be Ma Yongjun, 2001). At present the use of DDT is
redistributed on a global scale (Tanabe, 1991) .The still allowed to control mosquitoes, particularly in
origin and fate of OCPs in soils with different land the malarial transmission zones in China (Zhang et
use have been extensively studied in many al., 2005). Accordingly, China still produces a
countries. Although the usage of OCPs was phased small amount of DDT and China is also allowed to
out for decades, the elevated concentrations were export DDT to other countries for the same
still observed in many agricultural soils (Harris et purpose. This paper presents the current status of
al., 2000) and the relationship between sites of OCPs residues in Dayan cave (Karst cave).

35
Distribution and Sources of Organochlorine Pesticides (OCPs) in Karst Cave, Guilin, China
Annette Sylvie Muhayimana et al.
____________________________________________________________________________________________________________

2. MATERIALS AND METHODS

2.1 Study Area middle part of Guangming Mountain at right side

Region of research was in Guilin located in of Taohua River in the north-west of Guilin.
Guangxi Zhuang Autonomous Region in southeast Guangming Mountain is a large peak cluster in
China.Guangxi province (Southeast of China). The Fenglin Plain, with an area of 0.92km2, the highest
Geographical coordinates are 25° 40' 25" North, peak altitude of 404.4m and the plain altitude of
108° 44' 0" East and has an altitude of 150m. It is 151 m.The outcrop is a thick limestone layer of the
bounded to the north-east by Hunan province, to Devonian system with a high intensity of Karst
the south-east by Hezhou town and it is next to process. Dayan is a noncommercial karst cave
Guangdong province. It has a surface area of 27, located northeast to Ludiyan cave. The map of
800 square kilometers and a population of 4.76 Guangxi showing Guilin and plane diagram of
million. Dayan cave are shown in Fig 1 and Fig 2
respectively.
Dayan is an intermediate upper layer cave of
Guilin Maomoatou cave system, located in the

Ch ina
G uilin

Gua n gx i Zh u ang
Aut on o mo us Re gion

Fig 1: Map of Guangxi province showing Guilin

Fig 2: Map of Dayan Cave showing sampling locations

(1 to 10) are sampling locations

36
Marsland Press
Journal of American Science 2009: 5(1), 35-43
____________________________________________________________________________________________________________

2.2 Soil sampling the samples. Finally samples were stored and kept
Ten sampling locations were chosen inside the in the refrigerator until next analysis (Analysis by
cave that followed the horizontal section from the HP 6890 GC).
east gate as shown in Fig 2. Sampling location 1
was at the east gate (outside the cave) and the 2.3.2 Analysis by HP 6890 GC
serial number was from 1 to10. Three samples (1’, HP 6890 GC (Gas Chromatography) was
2’, and 3’) were also taken outside the east gate. equipped with a 63 Ni electron capture detector and
Nine samples were obtained inside the cave (2 a 30 m x 0.32 mm i.d (0.25 lm film thickness) DB-
to10) and 4 samples outside the cave (1, 1’, 2’, and 5 fused silica capillary column. Nitrogen was
3’). Sampling was done with the use of a hand added as a carrier gas at 1.2ml/min. the oven
shovel. The weight of each sample collected was temperature was kept at 40oC for 5 minutes and
500g. After the collection of samples, they were increased to 290oC at a rate of 4oC/min. Injector
kept frozen prior to the commencement of the and detector temperatures were maintained at 250
laboratory analysis. and 300oC respectively. 2 Microliters (µl) of each
sample was injected for analysis.
2.3 Analysis
2.3.1 Experimental procedures 2.3.3 Quality control and Quality assurance
Before analyising the samples (before (QC/QA)
experiment) all glass wares were acid washed and Quality control and Quality assurance was
cleansed with distilled water before they were made by the use of the US EPA method in the
dried in the oven at 200oC for about four hours. process of the experiment. Method blanks
Reagents used for the experiment included: (solvents), duplicate samples, and spiked blanks
dichloromethane (DCM), hexane, acetone, sodium (standards spiked into solvent) were analyzed. In
sulfate, alumina gel (100-200 mesh), silica gel addition, surrogate standards were added to each
(100-200 mesh), mesh hydrochloric acid and of the samples to monitor procedural performance
vitriol. Filter paper, aluminium foil, absorbent and matrix effects. The concentrations of OCPs
cotton and active copper were also used as were corrected for the recovery ratios for the
materials. surrogates. The recovery ratios for the surrogates
in the samples conform to the reported ranges by
Mixed standard sample of OCPs [2,4,5,6- US EPA. The recovery rates and standard
tetrachloro-m-xylene (TCMX) and deviation of OCPs during separation and testing
decachlorobiphenyl (PCB 209)] were used as are within the limiting value of the US EPA 610
surrogate standards and were added to all the method. Recovery rates of TCMX and PCB209 are
samples before the extraction. The whole process 69±6% and 76±7% respectively.
of pretreatment was based on US EPA SW-8080A
method as reference. 20 g of the sample were 3. RESULTS AND DISCUSSIONS
weighed with electronic balance and injected with
the surrogate (using a syringe) before the sample 3.1 Concentration and distribution of OCPs
was Soxhlet-extracted for 48 hours with redistilled A summary of concentrations of OCPs
Dichloromethane (DCM). Active copper slices detected in soil samples of Dayan cave is shown in
were added to the conical flask containing DCM to Table1.
eliminate the influence of sulphur contained in the Inside the cave ∑OCPs detected was 29.659 ng /g
with a mean value of 3.295 ng /g and ∑OCPs
sample. After 48 hrs in the soxhlet extractor, the detected outside the cave was 74.108 ng /g with a
extracted samples were added with Sodium mean value of 18.527ng/g. ∑OCPs outside the
sulphate (NaSO4) to remove unwanted water. After cave is higher than the total concentration outside
that, the solvents were concentrated to about 5 ml the cave (Fig 3).
and then passed though a mixture of silica gel and
alumina gel (10/3, V/V) for purification and it was The levels of OCPs outside the cave compared
rinsed by a mixture of DCM and hexane (2/3,V/V). to the levels inside indicated that despite the
relatively closed environmental system of the cave
The solvent was then condensed with high purity and less human interference inside the cave, it still
Nitrogen. 4 ml of the hexamethyl-benzene and had OCPs contamination due to air transfer, rain
PCNB (5ppb) were added as internal standards to water filtration and other processes, but the degree
help in quantifying the amount of OCPs present in of contamination was not high.

37
Distribution and Sources of Organochlorine Pesticides (OCPs) in Karst Cave, Guilin, China
Annette Sylvie Muhayimana et al.
____________________________________________________________________________________________________________

Table 1 Levels of OCPs in soil samples of Dayan Cave

OCPs overall level range OCPs level range inside the OCPs level range outside
Min─Max(mean value） cave the cave
Min─Max(mean value) Min─Max(mean value)
（2 to 10 samples） （1，1′，2′，3′ samples）
α-HCH 0.014─0.170 (0.087) 0.014─0.126(0.043) 0.095─0.170(0.130)
β-HCH 0.026─0.219 (0.102) 0.026─0.219(0.087) 0.100─0.138(0.117)
γ-HCH 0.015─0.285 (0.092) 0.015-0.180(0.044) 0.075─0.285(0.140)
δ-HCH 0.009─0.072 (0.034) 0.009─0.045(0.024) 0.020─0.072(0.044)
TC 0.021─6.119 (1.841) 0.021─1.674(0.279) 3.226─6.119(3.403)
CC 0.085─7.134 (2.221) 0.101─3.111(0.849) 3.899─7.134(4.198)
Hep ND─2.465 (1.399) ND─1.087(0.139) 1.871─2.465(1.632)
Hep-Epo ND─1.908 (0.911) ND─1.022(0.379) 1.000─1.908(1.185)
EndoI ND─0.230 (0.067) ND─0.040(0.021) 0.103─0.230(0.122)
EndoII ND─0.161 (0.046) ND─0.057(0.021) 0.026─0.161(0.080)
Endosulfate 0.030─0.500 (0.175) 0.030─0.180(0.086) 0.200─0.500(0.294)
P,p’-DDE 0.011─0.342 (0.108) 0.011─0.109(0.041) 0.115─0.342(0.174)
P,p’-DDD ND─0.121 (0.079) ND─0.077(0.038) 0.011─0.121(0.059)
O,p’-DDT 0.049─0.467 (0.212) 0.049─0.226(0.113) 0.302─0.467(0.310)
P,p’-DDT ND─0.090 (0.031) ND─0.039(0.011) 0.046─0.090(0.050)
ΣDDTsb 0.094─0.875 (0.371) 0.094─0.384(0.162) 0.532─0.875(0.434)
ΣHCHsa 0.100─0.665(0.269) 0.100─0.453(0.197) 0.313─0.665 (0.430)
ΣOCPsc 1.159─23.625(10.911) 1.159─11.180(3.295) 13.250─23.625(18.527)
ND=Non- detected
ΣHCHsa= α-HCH + β-HCH + δ-HCH + γ-HCH.
ΣDDTsb= p, p’-DDE + p, p’-DDD + o, p’-DDT + p, p’-DDT.
ΣOCPsc =ΣHCHs+ΣDDsT+Σother OCPs.
Σother OCPs = Heptachlor (Hep) + Heptachlor epoxide (Hep-Epo) + TC (Trans-Chlordane) + CC (Cis-Chlordane)
+EndoI (α- Endosulfan) +EndoII (β-Endosulfan) +Endosulfate.

The concentration of Chlordane (TC+CC) in suggests that there may be a fracture pore near the
OCPs was highest among all the OCPs detected north mouth that allows some air to come in.
inside and outside the cave with a total
concentration of 39.689ng/g and mean value of Fig 4 shows that the total concentration of
9.92 ng /g inside the cave and a total concentration DDTs (∑DDTs) in soil samples was higher than
of 4.52 ng/g outside the cave with a mean value of the total concentration of HCHs (ΣHCHs). This
1.13 ng /g. This is because South china have been trend is consistent with the previous observations
using Chlordane to kill termites, so the high on the contamination of OCPs in soil in China
concentration of Chlordane observed may be (Zhou et al., 2001). A most likely explanation for
predominantly due to the use of technical the current low concentration of HCHs in soil is
Chlordane as a termiticide in this area in previous due to the difference in the physicochemical and
years . In China, technical chlordane is still being biochemical properties, wherein HCHs have
extensively used against termites in buildings, with higher water solubility, vapor pressure and
an estimated amount of over 200 tons year−1 in biodegradability, and lower lipophicility and
recent years (Xu et al., 2004). particle affinity compared to the DDTs (Rui et al.,
The next compounds with highest levels of 2005). DDTs tend to remain in the particulate
concentration were Heptachlor (Hep) and phase longer than HCHs. (Nhan et al., 2001).
Heptachlor epoxide (Hep-Epo.) Heptachlor (Hep)
was also used and produced in large quantity in In comparison with recent research reports, the
China. From 1967 to 1969 the amount of concentrations of ∑DDTs and ∑HCHs measured
Heptachlor produced was 17 tons, to kill the in the study area were in the same low range with
termites and other insects in the soil. It is shown in other pristine areas such as Tibet plateau where the
Fig 4 that the majors parts of OCPs (HCHs and concentration of ∑DDTs ranged from ND to 2.83
DDTs) at the cave’s innermost sampling locations ng/g and ∑HCHs ranged from 0.18 to 5.38 ng/g
9 and 10 did not show the lowest values, but rather (Fu et al., 2001), and European high altitude
slightly greater than the values of sampling mountains that had ∑DDTs and ∑HCHs residual
locations 7 and 8 at the middle of the cave. This level in the range of 1.7-13 ng/g and 0.08-0.49
ng/g respectively (Grimalt et al., 2004).

38
 25

OCPs levels in soil (ng/g )

20
15
ΣOCPs
10
5
0
3' 2' 1' 1 2 3 4 5 6 7 8 9 10
Sampling locations

Fig 3: Distribution of ΣOCPs in soil of Dayan cave

1.6
1.4
DDTs and HCHs levels(ng/g)

1.2
1.0
0.8 ΣHCH
0.6 ΣDDT
0.4
0.2
0.0
3' 2' 1' 1 2 3 4 5 6 7 8 9 10
Sampling locations

Fig 4: Distribution of ΣHCHs and ΣDDTs in soil of Dayan cave

The average concentration outside the cave and HCH to γ-HCH has been used to identify the
inside the cave of ∑DDTs and ∑HCHs was lower possible HCH source. If the source of HCH comes
than the average concentration of ∑DDTs and from fresh input of technical HCH, the ratio of α -
∑HCHs in Hong Kong soils which was 0.52 ng/g to γ -HCH is between 3 and 7 (Yang et al., 2008).
and 6.19ng/g respectively (Zhang et al, 2006), and However, a lindane source will reduce the ratio to
they were much lower than the average close or <1 (Willet et al., 1998). A higher ratio of α
concentrations of ∑DDTs (37.6 ng/g) and ∑HCHs - to γ -HCH than 7 can be explained by long-range
(12.2 ng/g) found in soils of Pearl River Delta transport or re-cycling of technical HCH, because
Region (Fu et al., 2003). Some other studies α -HCH has a longer atmospheric lifetime than γ
reported around China, had higher residual levels isomer by about 25% (Willet et al., 1998). As
of OCPs such as Beijing (Zhu et al, 2005), Tianjin shown in Fig 6, the ratios of α-HCH/γ-HCH in all
(Tao et al., 2005), Nanjing (An et al., 2005). In soil sampling locations were lower than 3.
Europe, ΣDDTs and ΣHCHs levels were in the Accordingly, the contamination of HCHs in this
range of 4.3-2400 ng/g and 0.36-110 ng/g in region probably came from local use of lindane
Poland soils (Falandysz et al., 2001). In and also indicated Lindane inputs in the past
comparison with similar research the levels of several years. By analyzing the individual HCH
OCPs in Guilin were low and the reason is because isomers (Fig 5), it was found that β-HCH had the
there are mainly rice farms in the vicinity of Guilin highest level of concentration among all the
city in which small amounts of OCPs were used samples and it accounted from 20.03-79.13 %,
with the rotary method of planting rice. The especially in sample 3 to 7 where it accounted
existence of alternating wet and dry conditions was from 23-79% of the total HCHs detected. The β-
beneficial to the aerobic and anaerobic degradation HCH was higher because of its persistence in soil.
of OCPs, leading to a reduced amount of soil The persistence of β-HCH in soils is mainly due to
OCPs. the higher Kow (log Kow =3.78) and lower vapor
pressure value (3.6x10-7 mmHg, 20oC) (Zhang et
3.1.1 Distribution and degradation of HCH al., 2006). These will make β-HCH easier to be
isomers absorbed to the soil organic matter and less
It has been widely recognized that HCH is evaporative loss from soils (Mackay et al., 1997).
available in two formulations: technical HCH and Furthermore, the spatial arrangement of Chlorine
lindane. Technical HCH contains isomers in the atoms in the molecular structure of β-HCH was
following percentages: α, 55–80%; β, 5–14%; γ, 8– supposed to be more resistant to microbial
15%; δ, 2–16%; ε, 3–5% (Qiu et al., 2004), and degradation in soils (Middeldorp et al., 1996).
Lindane contains > 90% of γ-HCH. The ratio of α-

39
Distribution and Sources of Organochlorine Pesticides (OCPs) in Karst Cave, Guilin, China
Annette Sylvie Muhayimana et al.
____________________________________________________________________________________________________________

0.3

HCH isomers (ng/g)

a-HCH
0.2 b-HCH
g-HCH
0.1 d-HCH

0.0
3' 2' 1' 1 2 3 4 5 6 7 8 9 10
Sampling locations

Fig 5: HCH isomers in soil of Dayan cave

2.5
αHCH/γHCH
a-HCH/r-HCH

2
1.5
1
0.5
0
3' 2' 1' 1 2 3 4 5 6 7 8 9 10
Sampling locations

Fig 6: Ratios of α-HCH to γ-HCH in soil of Dayan cave

3.1.2 Distribution and degradation of DDT environment. The ratio of (DDE+DDD)/ΣDDTs

isomers greater than 0.5 can be thought to be subjected to a
Commercial DDT generally contains 75% long term weathering (Dong et al, 2002) and More
p,p’-DDT, 15% o,p’-DDT, 5% p,p’-DDE, <0.5% o, p′-DDT than p, p′-DDT in the environment can
p,p’-DDD, <0.5 o,p’-DDE and <0.5% of demonstrate the Dicofol type DDT usage (Qiu et
unidentified compounds (WHO, 1979), but in al., 2004 ).
Dicofol, the concentration of o,p’-DDT is more
than p,p’-DDT (Qiu et al., 2005). DDTs isomers The ratios of (DDE+DDD)/ ΣDDTs are shown
have a long persistence in the environment and in Fig.9. The ratios were in the range of 0.26-0.61
their levels of concentrations in this study are with most values being less than 0.5 (mean value
shown in Fig.7. DDT can be biodegraded under is 0.4) and in Fig.7 it is shown that the
aerobic conditions to DDE and under anaerobic concentration of o,p’-DDT was more than the
conditions to DDD (Bossi et al., 1992). The ratio concentration of p,p’-DDT as in Dicofol, this
of DDD/DDE greater than 1 indicates that the soil suggests that there was fresh input of Dicofol in
was dominated by DDD, the product of anaerobic the study area. Also, most values of DDD/DDE
degradation of DDT, and the ratio lesser than 1 ratios as shown in Fig. 8 were greater than 1 inside
indicates that the soil was dominated by DDE, the the cave and ranged from 0.092 to 7 with an
product of aerobic degradation of DDT (Zhou et average value of 2.31, and the ratios of DDD/DDE
al., 2006). DDE and DDD Changes in the ratio of outside the cave ranged from 0.052 to 0.53 with an
DDE and DDD to ΣDDTs has been regarded as an average value of 0.35.
indication of either no or decreasing inputs to the

40
 p,p’-DDE

( ng/g)
0.5
p,p’-DDD
0.4 o,p’-DDT

DDT iso mers levers

0.3 p,p’-DDT

0.2

0.1

0.0
3' 2' 1' 1 2 3 4 5 6 7 8 9 10
Sampling locations

Fig 7: Distribution of DDT isomers in soil of Dayan Cave

The results obtained clearly indicated that DDT in The use of Dicofol in China is mainly in the
soil inside and outside of the Dayan cave may be southern and eastern provinces, mostly on litchi,
derived from Dicofol and DDT was retained under longan, citrus crops and cotton (Yang et al., 2008).
anaerobic conditions inside the cave and under
aerobic condition outside the cave.

8
7
6
DDD/DDE

5
4
3 pp'-DDD/pp'-DDE
2
1
0
3' 2' 1' 1 2 3 4 5 6 7 8 9 10
Sampling locations

Fig 8: Ratios of DDD/DDE in soil of Dayan Cave

41
Distribution and Sources of Organochlorine Pesticides (OCPs) in Karst Cave, Guilin, China
Annette Sylvie Muhayimana et al.
____________________________________________________________________________________________________________

1.0

0.8

(DDD+DDE)/DDTs
( DDD+DDE) /ΣDDTs
0.6

0.4

0.2

0.0
3' 2' 1' 1 2 3 4 5 6 7 8 9 10

Sampling locations

Fig 9: Ratios of (DDD + DDE)/ ΣDDTs in soil of Dayan cave

4. CONCLUSION

The use of HCHs and DDTs in China has been Qi Shihua

banned for 20 years and this sanction resulted in a Department of Environmental Engineering
tremendous decrease of OCPs concentrations in Faculty of Environmental Studies,
soils of Dayan cave. The residual levels of OCPs Key Laboratory of Bio and Environmental
in soils outside Dayan cave were less than Geology of Ministry of Education,
corresponding national values and among all the China University of Geosciences, Wuhan, Hubei
OCPs detected the concentration of chlordane and 430074, China
heptachlor were highest because they have been E-mail: shihuaqi@cug.edu.cn
used in the study area. ΣDDTs and ΣHCHs in soil
inside the cave were low in comparison with
worldwide background mountains and polar REFERENCES
regions. As conclusion the pollution of OCPs in
the soils inside and outside Dayan cave was light. 1. An, Q., Dong, Y.H., Wang, H., 2005.
The analysis of isomers of DDTs and HCHs Residues and distribution character of
showed that there is fresh use of Dicofol and organochlorine pesticides in soils in Nanjing
Lindane respectively in the study area. DDT area. Acta Scientiae Circumstantiae, 25, 470-
degradation outside the cave was aerobic while 474.
inside the cave the degradation of DDT was 2. Bossi, R., Laesen, B., Premazze, G., 1992.
anaerobic. Polychlorinated biphenyl congeners and
other chlorinated hydrocarbons in bottom
ACKNOWLEDGEMENT sediment cores of lake Garden (Italy), Sci.
Total Environ., 121, 77–93.
The authors wish to pay tribute to the 3. Colborn, T., Dumanoski, D., Myers, J.P.,
referenced authors. This study discusses their work, 1996. Our Stolen Future. Dutton, New York,
but the views expressed are those of authors and USA.
do not represent those of the referenced authors. 4. Doong, R.A., Peng, C.K., Sun, Y.C., Liao,
P.L., 2002.Composition and distribution of
Corresponding to: organochlorine pesticide residues in surface
Annette Sylvie Muhayimana sediments from the Wu-Shi River estuary,
Department of Environmental Engineering Taiwan. Mar. Pollut. Bull., 45, 246–253.
Faculty of Environmental Studies, 5. Falandysz, J., Brudnowska, B., Kawano M.,
Key Laboratory of Bio and Environmental 2001. Polychlorinated Bi-phenyls and
Geology of Ministry of Education, Organochlorine Pesticides in Soils from the
China University of Geosciences, Wuhan, Hubei Southern Part of Poland. Arch. Environ
430074, China Contam. Toxicol., 40:173-178
E-mail: teteli2001@yahoo.com

42
6. Fu, J.M., Mai, B.X., Sheng , G.Y., Zhang, G., Sediments collected from East China Sea.
Wang, X.M., Peng, P.A., Xiao, X.M., Ran, Environmental international, 31,799-804.
R., Cheng, F.Z., Peng, C.Z., Wang, Z.S., 18. Shivaramaiah, H.M., Odeh, I.O.A., Kennedy,
Tang, U.W., 2003. Persistent organic I.R., Skerritt, J.H., 2002. Mapping the
pollutants in environment of the Pearl River distribution of DDT residues as DDE in the
Delta, China: an overview. Chemosphere, 52, soils of the irrigated regions of Northern
1411-1422. New South Wales, Australia using ELISA
7. Fu, S., Chu, S.G., Xu, X.B., 2001. and GIS. J. Agric. Food Chem., 50, 5360–
Organochlorine pesticide residue in soils 5367
from Tibet, China. Bull. Environ. Contam. 19. Tanabe, S., 1991. Fate of toxic-chemicals in
Toxicol., 66, 171–177 the tropics. Marine Pollution Bulletin, 22,
8. Grimalt, J .O., Drooge, B .L., Ribes, A., 259–260.
Vilanova, R. M., Fernandez, P., Appleby, P., 20. Tao, S., Xu, F.L., Wang, J., Liu, W.X.,
2004. Persistent organochlorine compounds Gong, Z.M., Fang, J.Y., Zhu, L.Z., Y.M.
in soils and sediments of European high Luo, Y.M., 2005.Organochlorine pesticides
altitude mountain lakes. Chemosphere, 54 in agriculture soil and vegetables from
(10): 1549 Tianjin, China. Environ. Sci. Technol., 39,
9. Harris, M.L., Wilson, L.K., Elliott, J.E., 2494–2499.
Bishop, C.A., Tomlin, A.D., Henning, K.V., 21. Willett, K.L., Ulrich, E.M., Hites, S.A., 1998.
2000. Transfer of DDT and metabolites from Differential toxicity and environmental fates
fruit orchard soils to American Robins of Hexachlorocyclohexane isomers. Environ.
(Turdus migratorius) twenty years after Sci. Technol., 32(15): 2197-207
agricultural use of DDT in Canada. Arch. 22. World Health Organization (WHO), 1979.
Environ. Contam. Toxicol., 39, 205–220. DDTs and its derivations. WHO, New York.
10. Mackay, D., Shiu, W.Y., Ma, K.C., 1997. 23. Xu, D., Deng, L., Chai, Z., Mao, X., 2004.
Illustrated Handbook of Physical-Chemical Organohalogenated compounds in pine
properties of Environmental Fate of Organic needles from Beijing city, China.
Chemicals, vol.V. Lewis Publishers, Boca Chemosphere, 57, 1343–1353.
Raton, FL. 24. Yang, Y., Li, D., Mu, D., 2008. Levels
11. Meijer, S.N., Halsall, C.J., Harner, T., Peters, seasonal Variations and sources of OCPs in
A.J., Ockenden, W.A., Johnston, A.E., Jones, ambient air of Guangzhou, China.
K.C., 2001. Organochlorine pesticides Atmospheric Environment, 42, 677-687.
residues in archived UK soil. Environ. Sci. 25. Zhang, H.B., Luo, Y.M., Zhao, Q.Z., Wong,
Technol., 35, 1989–1995. M.H., Zhang, G.L., 2006. Residues of
12. Middeldorp, P.J.M., Jaspers, M., Zehnder, Organochlorine pesticides in Hong Kong
A.J.B., Schraa, G., 1996. Biotransformation soils. Chemosphere, 63, 633-641.
of α-, β-, γ-, and δ- hexachlorocyclohexane 26. Zhang, H., Lu, Y., Dawson, R.W., Shi, Y.,
under methanogenic conditions. Environ. Wang, T., 2005. Classification and
Sci.Technol., 30, 2345-2349. ordination of DDT and HCH in soil samples
13. Negoita , T .G., Covaci ,A., Gheorghe, A., from the Guanting Reservoir, China.
Schepens, P. , 2003.Distribution of Chemosphere, 60, 762-769.
polychlorinated biphenyls (PCBs) and 27. Zhao, L., Ma, Y., 2001. Effect of organic
organochlorine pesticides in soils from the residues on soil environment. Soils, 33, 309-
East Antarctic coast. Journal of 311.
Environmental Monitoring, 5 (2):281-286. 28. Zhou, J.L., Maskaoui, K., Qiu, Y.W., Hong,
14. Nhan, D.D., Carvalho, F.P., Am N.M., Tuan, H.S., Wang, Z.D., 2001.Polychlorinated
N.Q., Yen, N.T.H., Villeneuve, J.P., 2001. biphenyl congeners and Organochlorine
Chlorinated pesticides and PCBs in insecticides in the water column and
sediments and molluscs from fresh water sediments of Daya Bay, China. Environ
canals in the Hanoi region. Environ. Pollut., Pollut., 113: 373-84
112, 311-20. 29. Zhou, R., Zhu, L., Yang, K., Chen, Y., 2006.
15. Qiu, X.H., Zhu, T., Li, J., Pan, H.S., Li, Q.L., Distribution of OCP’s in surface water and
Miao, G.F., Gong, J.C., 2004. Sediments from Qiantang River, East China.
Organochlorine pesticides in the air around Department of Environmental Science,
the Taihu Lake, China. Environ. Sci. Zhejiang University.
Technol., 38, 1368-1374. 30. Zhu, Y., Liu, H., L., Xi, Z., Cheng, H., Xu,
16. Qiu, X.H., Zhu, T., Yao, B., Hu, S.W., 2005. X., 2005.Organochloline pesticides (DDTs
Contribution of Dicofol to current DDT and HCHs) in soils from outskirts of Beijing,
pollution in China. Environ. Sci. Technol., China. Chemosphere, 60, 700-778.
39, 4385-4390.
17. Rui, Q.Y., Gui, B.J., Qun, F.Z., Chun, G.Y,
Jian, J.B.S., 2005. Occurrence and 3/1/2008
distribution of Organochlorine pesticides in

43
Marsland Press
Journal of American Science 2009: 5(1), 44-48
___________________________________________________________________________________________________
Research Article

An Incidence of Substratum Discolouration in a Tropical West African Lagoon.

Onyema, I.C. and Nwankwo, D.I.
Department of Marine Sciences, University of Lagos, Nigeria.
iconyema@gmail.com
_____________________________________________________________________________________________
ABSTRACT: A greenish discolouration of the lagoon floor at the Bayeku area of the Lagos lagoon was observed
in January 2006. We report here an investigation of the area between December, 2005 and Feburary, 2006 as part of
a larger study. A total of 19 species from 13 genera were reported. Oscillatoria tenuis (95,800 trichomes per ml) was
implicated as the causative organism for the substratum discolouration. Increased insolation, especially reaching the
lagoon floor, low salinity, absence of flood conditions, suitable sediment type (fine – medium sand) and high
nutrient (PO4 - P > 0.24 mg/L; NO3 – N > 4.40mg/L) levels possibly encouraged the algal proliferation and
subsequent substratum discoloration. It is suggested that improving water quality indices and salinity after January
caused the disappearance of the discolouration on the substratum. [Journal of American Science 2009: 5(1), 44-48]
(ISSN: 1545-1003)

Key words: algae, water quality indices, substratum.

1. INTRODUCTION period and after the collapse. Water quality indices before,
Coastal algal blooms respond to nutrient load from during and after the substratum discolouration were also
anthropogenic sources (Lee, 1999; Onyema, 2007). South- estimated and investigated. This report is part of a larger
western Nigeria is endowed with an intricate network of study that was already ongoing at the time of the
rivers, creeks and lagoons, that serve as conduits occurrence.
transferring highly nutrified waters from hinterland to
coastal areas. Bloom conditions have been reported in
2. MATERIALS AND METHODS
some of these waters (Nwankwo et al., 2003a; Nwankwo
et al., 2008). Blooms of Microcystis aureginosa, M. flos- Description of study area:
aquae and M. wesenbergii were reported in the Lagos
The Lagos lagoon opens into the sea via the
lagoon (Nwankwo, 1993), Ogun river at Iju (Nwankwo,
Lagos habour all through the year. The tidal height is low
1993) causing bluish colouration, anoxia, odour, impacting
(<1.5m) and the tidal exchange weak. It is shallow (<2m)
taste to the water (Nwankwo et al., 2003a) and kuramo
and connected to the Epe lagoon to the east. The area
lagoon (Nwankwo et al., 2008). Blooms of Trichodesmium
investigated was (Fig 1) the Bayeku area of the Lagos
thiebautii have also been reported off the Lagos coast
lagoon (Latitudes 60 32lN and 60 31lN and Longitudes 30
(Nwankwo, 1993) during thermocline conditions and more
31lE and 30 32lE). A greenish, slimy covering of suspected
recently a bloom of Bellerochea malleus that caused
algae on the lagoon floor was observed for the very first
brownish discolouration off the Light house beach, Lagos
time in this area. Nutrient rich water is known to flow
(Nwankwo et al., 2004) was documented. Blooms of
from eutrophic creeks and creeklets systems in the area.
Anabaena flos-aquae, A. spiroides (cyanobacteria),
Furthermore, poor sewerage systems are the common state
Cerataulina bergoni, Chaetoceros convolutus,
of the rural dwellers of the immediate area. Hence direct
Coscinodiscus centralis (diatoms) and Ceratium furca, C.
dumping of domestic wastes is carried out in the closet
fusus, C. tripos and Noctiluca scntillans (dinoflagellates)
water body.
are known to induce harmful effects in waters of south-
western Nigeria (Nwankwo, 1993; Nwankwo et al., 2003a,
Collection of samples
b, Onyema, 2008). There is at present a report of
substratum discolouration in the Lagos lagoon system
Water samples for determining water quality
(Onyema and Nwankwo, 2006) implicating Beggiatoa
characteristics were collected at the site before substratum
alba and Oscillatoria spp as causative species.
sample collection. The boat was anchored throughout
sample collections. Water samples were collected in 1L
Between December, 2005 and Feburary, 2006, a
plastic bottles with screw cap from 0.5m depth from the
greenish discolouration of the substratum at Bayeku was
water surface. This was labeled and transported to the
observed and thoroughly investigated. We report here the
laboratory for chemical analysis.
composition of the organisms before, during the bloom

44
An Incidence of Substratum Discolouration in a Tropical West African Lagoon.
Onyema, I.C. et al.
___________________________________________________________________________________________________________________________

Substratum samples (top 5cm) were collected were collected on each occasion. Out of water and in the
within a 5cm² quadrat carefully placed on the greenish boat, samples were transferred to 75cl screw capped
material / lagoon floor. A spatula was gently used plastic containers. Samples were fixed with formalin (4%
underwater to scrape the topmost part. After carefully unbuffered) and labeled appropriately on the field before
scooping up the greenish scum, it was gently spooned into onward transportation to the laboratory. This process was
a plastic bag while still underwater. Duplicate samples carried out on each sampling occasion.

Physico-chemical analysis nitrogen values while sulphate levels were measured using
Air and surface water temperatures were the gravimetric method. Calcium and magnesium ions
measured in-situ using a mercury thermometer while water were determined using a 400 single channel, low flame
depth was estimated with a calibrated pole. Total dissolved photometer. Concentrations of copper, iron and zinc were
solids was determined by evaporating 100ml aliquot at determined with an atomic absorption spectrophotometer
105oC and total suspended solids estimated by filtering (A.A.S.) Uni cam 99model.
100ml of sample through a pre-weighed filter paper, dried
to constant weight and reweigh. Conductivity was
Biological Analyses
measured using the HANNA instrument while salinity was
In the laboratory, the drop count microscope
determined using the silver-nitrate chromate method. The
analysis method described by Onyema (2007) was used to
surface water pH was determined with a Griffin pH meter
estimate the substratum algal flora. Microscope analysis
(Model 80) while dissolved oxygen was measured using a
was carried out on samples within 48hours of collection.
Griffin oxygen meter (Model 40). Biological and chemical
Identification materials were used to assist and confirm
oxygen demands were measured using methods described
identification of species (Smith 1950; Hendey, 1958, 1964;
in APHA (1998) for water analysis. Calorimetric methods
Desikachary, 1959; Wimpenny, 1966; Patrick and Reimer,
using a Lovibond Nesslerier were adopted for the direct
1966, 1975; Whitford and Schmacher, 1973;
determination of phosphate-phosphorus and nitrate-

45
Marsland Press
Journal of American Science 2009: 5(1), 44-48
___________________________________________________________________________________________________
Vanlandingham, 1982; Nwankwo, 1990, 2004a; Bettrons - 11mg/L) and chemical oxygen demands (10 – 49 mg/L)
and Castrejon, 1999; Siver, 2003; Rosowski, 2003). and heavy metals levels (Iron 0.14 - 0.35, Zinc 0.003 -
0.006mg/L) (Table 1).

3. RESULTS With regard to the algae, just one species each

Physico-chemical was recorded for December 2005 (Microcystis aureginosa
Air (31 - 32 oC) and water (30 - 31 oC) Kutzing) and January 2006 (Oscillatoria tenuis Agardh),
temperatures were high through out the sampling period However, 17 species were recorded in February (Table 2).
while the sampling depth was averagely 1.31m. The water Although, total biomass in terms of cell numbers was high
remained slightly alkaline throughout the study (7.01 – in January (95,800 trichomes per ml) it was for a sole
7.10). The total dissolved solids (20 - 33 mg/L), salinity species. This organism (Oscillatoria tenuis Agardh) is the
(2.30 - 20.60 o/oo), chloride content (770.0 – 6930 mg/L), implicated microalgae responsible for the greenish
conductivity (2335 – 12,500 µS/cm), acidity (3.0 - 8.8 discolouration of the lagoon floor at Bayeku. Furthermore,
mg/L), alkalinity (28.5 - 100.3 mg/L), total hardness February recorded 3 cyanobacteria, 8 centric diatoms and
(562.5 - 4687.0 mg/L), sulphate (6.1 – 60 mg/L) and 6 pennate diatoms species. Actinophycus splendens Ralfs
cation content (Calcium 111- 500, Magnesium 35.6- 859 and Biddulphia laevis Ehrenberg were important diatoms
mg/L) increased as the dry season progressed, while there and Oscillatoria limnosa Agardh for the cyanobacteria in
was a corresponding decrease in total suspended solids terms of numbers in February.
(1590 – 8260 mg/L), nitrate (2.5 - 4.8 mg/L), biological (5

46
An Incidence of Substratum Discolouration in a Tropical West African Lagoon.
Onyema, I.C. et al.
___________________________________________________________________________________________________________________________

4. DISCUSSION 2004b; Onyema, 2008). In Nigeria, Onyema et al., (2003)

The water quality status at the site ranged between low has reported Oscillatoria tenius in organically polluted
and high brackish water conditions. Low brackish parts of Lagos lagoon. Similarly, Chindah and Pudo (1991)
condition (S=2.30‰) was experienced in December while have reported Oscillatoria tenius from the Bonny river
high brackish condition (>9.20‰) reflected the dry months. associated with oil related effluent. According to
As the rain ceased, turbidity reduced while transparency Valangdiham (1982) Oscillatoria species are heavily
increased. Furthermore, insolation increased probably favoured in organically nutrified waters. The existence of
reaching the lagoon floor. This coupled with high nutrient high BOD levels in excess of 9mg/L at this site may be
levels (PO32- > 0.24mg/L, NO3- > 4.4mg/L, SO42- pointer to the probably stressed water quality status.
>6.1mg/L), low brackish condition (<9.20/00) and low According to Hynes (1960), BOD above 8.0mg/L may
depth (<1.42m), favorable sediment type (fine – medium indicate severe organic pollution. The disappearance of the
sand) and absence of flood conditions probably bloom in February may be associated with increased
encouraged the proliferation of the epipelic algal salinity (•20.60/00) and reduced nutrient load (PO4 – P
population in January. According to Valangdiham (1982), 0.04mg/L; NO3-N = 2.05mg/L). Onyema and Nwankwo
Oscillatoria tenius, the causative cyanobacterium, in the (2006) reported a high abundance of epipelic algal forms
substratum discolouration, is a saprobiont which can exist in the dry months at some organically polluted sites of an
either as plankton or as an attached form. Palmer (1969) estuarine creek in Lagos. This investigation highlights the
reported that Oscillatoria tenuis is the second most bane of increasing levels of pollutants from anthropogenic
tolerant Oscillatoria species to organically induced stress. sources in the Lagos lagoon and the role of algal indicators
It’s important to note that both sole species in December in capturing changes in water quality.
and January are known pollution tolerant cyanobacteria
forms for the region (Nwankwo, 2004b). Importantly, the Corresponding author:
highest level of nitrate (4.8 mg/L) recorded for this study Onyema, I.C.
was in January at the time of the greenish occurrence. Department of Marine Sciences,
Oscillatoria spp are reported in literature to have wide University of Lagos, Nigeria.
tolerance limits to pH, salts and organically enriched iconyema@gmail.com
environments (Valangdiham, 1982; Lee, 1999; Nwankwo,

47
Marsland Press
Journal of American Science 2009: 5(1), 44-48
___________________________________________________________________________________________________
REFERENCES 16. Onyema, I.C. (2007). Mudflat microalgae of a tropical
1. American Public Health Association. (1998). bay in Lagos, Nigeria. Asian Journal of Microbiology,
Standard Methods for the Examination of Water and Biotechnology and Environmental Sciences. 9 (4):
Waste Water. 20th ed. APHA New York. 1270pp. 877 – 883.
2. Bettrons, D.A.S. and Castrejon, E.S. (1999). 17. Onyema, I.C. (2008). Phytoplankton biomass and
Structure of benthic diatom assemblages from a diversity at the Iyagbe lagoon Lagos, Nigeria.
mangrove environment in a Mexican subtropical University of Lagos, Akoka. Department of Marine
lagoon. Biotropica. 31(1): 48 – 70. Sciences. 266pp
3. Chinda, A.C and Pudo, J. (1991). A preliminary 18. Onyema, I.C. and Nwankwo, D.I. (2006). The epipelic
checklist of algae found in plankton of Bonny River in assemblage of a polluted estuarine creek in Lagos,
Niger Delta, Nigeria. Fragm flor. Geobot.36 (1): 112- Nigeria. Pollution Research. 25(3): 459 - 468.
126 19. Onyema, I.C., Otudeko, O.G. and Nwankwo, D.I.
4. Desikachary, T.V. (1959). Cyanophyta. Indian (2003). The distribution and composition of plankton
Council of Agric. Research, New Delhi. 686pp. around a sewage disposal site at Iddo, Nigeria.
5. Hendey, N.I. (1958). Marine diatoms from West Journal of Scientific Research Development. 7: 11-24.
African Ports. Journal of Royal Microscopic Society. 20. Smith, G.M. (1950). The fresh-water algae of the
77: 28-88. Hendey, N.I. (1964). An introductory United States. McGraw-Hill, London.719pp
account of the smaller algae of British coastal waters. 21. Siver, P.A. (2003). Synurophyte algae. In: Freshwater
Part 5. Bacillariophyceae (diatoms) London. Algae of North America. Ecology and Classification.
N.M.S.O. 317pp. Wehr, J.D. and Sheath, R.G. (Eds). Academic Press,
6. Hynes, H.B. (1960). The biology of polluted waters. New York. pp 523 – 558
Liverpool University Press, Liverpool, 1-202 pp 22. Palmer, C.M. (1969). A composite rating of algae
7. Lee, R.E. (1999). Phycology. Cambridge University tolerating organic pollution. Journal of Phycology.
Press, New York. 614pp. 5(1): 78 – 82.
8. Nwankwo, D.I. (1993). Cyanobacteria bloom species 23. Patrick, R. and Reimer, C.W. (1966). The diatoms of
in coastal waters of South-Western Nigeria. Archiv the United States exclusive of Alaska and Hawaii (Vol.
Hydrobiologie Supplement. 90: 553-542. 1). Monogr. Acad. Nat. Sci. Philadelphia. 686pp.
9. Nwankwo, D.I. (1990). Contribution to the Diatom 24. Patrick, R. and Reimer, C.W. (1975). The diatoms of
flora of Nigeria. Diatoms of Lagos lagoon and the the United States exclusive of Alaska and Hawaii (Vol.
adjacent sea. Nigerian Journal of Botany. 3: 53-70. 2, part 1). Monogr. Acad. Nat. Sci. Philadelphia.
10. Nwankwo, D.I. (2004a). A Practical Guide to the 213pp.
study of algae. JAS Publishers, Lagos. Nigeria. 84pp. 25. Rosowski, J.R. (2003). Photosynthetic Euglenoids. In:
11. Nwankwo, D.I. (2004b). The Microalgae: Our Freshwater Algae of North America. Ecology and
indispensable allies in aquatic monitoring and Classification, Wehr, J.D. and Sheath, R.G. (Eds).
biodiversity sustainability. University of Lagos Press. Academic Press, New York. pp 383 – 422.
Inaugural lecture seris. 44pp. 26. Round, F.E. (1953). An investigation of two benthic
12. Nwankwo, D.I., Onyema, I.C. and Adesalu, algal communities in Malham Tarn, Yorkshire. J. Ecol.
T.A.(2003a). A survey of harmful algae in coastal 41: 174-187.
waters of south-western Nigeria. Journal of Nigerian 27. Vanlandingham, S.L. (1982). Guide to the
Environmental Society. 1(2). 241 – 246. identification and environmental requirements and
13. Nwankwo, D.I., Onyema, I.C., Adesalu, T.A., pollution tolerance of freshwater blue-green algae
Olabode, R.J., Osiegbu, G.O. and Owoseni, T.I. (cyanophyta). U.S. Environmental Protection Agency,
(2003b). Additions to a Preliminary Checklist of EPA – 60.
Planktonic Algae in Lagos lagoon, Nigeria. Journal of 28. Whitford, L.A. and Schmacher, G.H. (1973). A
Science Technology and Environment. 3(1): 8 – 12. manual of freshwater algae. Sparks press Raeigh.
14. Nwankwo, D.I., Onyema., I.C, Labiran, C.O., 324pp.
Otuorumo, O.A.; Onadipe, E.I.;, Ebulu, M.O. and 29. Wimpenny, R.S. (1966). The plankton of the sea.
Emubaiye, N. (2004). Notes on the observations of Faber and Faber Limited, London. 426pp.
brown water discolouration off the light house beach,
Lagos, Nigeria. Discovery and Innovation. 16 (3).
111 – 116.
15. Nwankwo, D.I., Owoseni, T.I., Usilo, D.A., Obinyan
I., Uche, A.C. and Onyema, I.C. (2008).
Hydrochemistry and plankton dynamics of Kuramo
lagoon. Life Science Journal. 5 (3): 50 – 55.

48
ˠ˴̅̆˿˴́˷ʳˣ̅˸̆̆ʳ
˝̂̈̅́˴˿ʳ̂˹ʳ˔̀˸̅˼˶˴́ʳ˦˶˼˸́˶˸ʳ˅˃˃ˌˍʳˈʻ˄ʼʿʳˇˌˀˈˇʳ
˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲ʳ
Research Article
The Influences of Extremely Low Frequency AC Magnetic Fields At 60Hz on
Mung Beans Growth

Pai-Tsun Tien and Show-Ran Wang

Department of Electronic Engineering, National Taiwan University of Science and Technology, Taipei
106, Taiwan, ROC
E-mail:tyanbt@cht.com.tw
___________________________________________________________________________________
ABSTRACT: There are many reports about the biological effects of extremely low frequency magnetic
fields (ELF MFs),but few of them investigate how different intensity MFs act upon the growth of living
organisms. This study aims to assess the influences of the different intensity of ELF MFs on the early
growth of living organisms using mung beans as test materials. We used 60Hz 110Vrms AC electric
power as the source and made a toroidal magnetic coil by self for this experiment. The ELF MF is induced
using a magnetic circuit with a toroidal magnetic coil and a 60W lamp in series, which is driven by 60Hz
110V AC electric power, the maximum intensity of ELF MF is 950mG. To utilize the magnetic field
intensity decay when distance increase, to choose the three kinds different magnetic field intensity (such
as 875mG,155mG and 1.8mG rms value). We used three groups of mung beans (each group is 50 beans)
were exposed to the three kinds different magnetic field intensity separately, and observed the lengths of
stems and leaves of mung beans after five days growth. The results indicate that the magnetic field
intensity is 875mG and 155mG have an enhancing effect on the early growth of mung beans. [Journal of
American Science 2009: 5(1), 49-54] ʻ˜˦˦ˡˍʳ˄ˈˇˈˀ˄˃˃ˆʼ
Key words: ELF MF; biological effect,AC electric power,mung bean

1. INTRODUCTION to 10cm away). Because most countries adopt the

Because popularization of electricity and reference levels which were announced by ICNIRP in
modernization of life, to place in the electric power 1998 for general public exposure to time-varying
line generally and use home electrical appliances electric and magnetic fields as the standard. The
frequently on the human inhabitancy space, there are formula of reference level for general public is 50/f (f
ELF MFs produced also exists around the living is the frequency ,unit:KHz), the reference level is
space. We used a magnetic meter (TES-1390 ELF 833mG when f is 60Hz. For understanding the
Magnetic Field Meter, Bandwidth:50~300Hz,TES biological effect of different kinds magnetic field
Electrical Electronic Crop. made in Taiwan) to intensity, we made a toroidal magnetic coil by self,
measure the root mean square value of ELF MF the coil produced the maximum ELF MF intensity is
intensity of home electrical appliances such as 950mG. To utilize the magnetic field intensity decay
hairdryer, desk lamp, razor, etc. We can get magnetic when distance increase, to choose the three kinds
field intensity greater than 100mG (rms value), when different magnetic field intensity (such as
to measure home electrical appliances closely (5cm 875mG,155mG and 1.8mG rms value). We exposed
ʳ 49
The Influences of Extremely Low Frequency AC Magnetic Fields At 60Hz on Mung Beans Growth
Pai-Tsun Tien et al.
_________________________________________________________________________________________________________________
test materials (mung beans) in the three kinds three groups that were maintained in the test room
different magnetic field intensity, and observed were almost the same, and the light was supplied by
different magnetic field intensities act upon the early white fluorescent lamps .The close environmental
growth of test materials. parameters of three groups can be achieved so that
the growth difference between them only comes from
2. MATERIALS AND METHODS the magnetic field variable. The environmental
2.1 Plant material parameters such as temperature is 28±2ʚ, humidity
Mung beans were used as the test subject in this study. is 60±6 ʝ ,illumination is 1120±50LUX(day) and
We selected 150 mung beans of almost the same 563LUX(night).
weight (0.09 g) and similar appearance, so that the 2.2 Exposure System
sample error can be greatly reduced, and divided into The purpose of this study is mainly to assess the
three groups of 50 mung beans. Two groups of them influence on the early growth of mung beans exposed
are grown in a magnetic field (exposed group 1 under to the different magnetic field intensities. The
higher magnetic intensity and exposed group 2 under equipment needed in this experiment included a
lower magnetic field intensity), and the other group is 60-Watts incandescent lamp, a toroidal magnetic coil,
placed in an ambient weak magnetic field (control an oscilloscope/ frequency analyzer, etc. In order to
group). We used a rectangular culture plate produce the environment of higher magnetic intensity,
(dimension is 48Ø27Ø 3.5cm) which was spread the we made a toroidal magnetic coil with air gap by self
fine sand of depth 3cm to grow three groups of mung is shown in Figure 1.
beans together. The environmental parameters of

Fig 1: The toroidal magnetic coil with air gap

We entwined Iron wire (cross-section diameter
=2mm) to become a toroidal iron core with diameter Ni lc l
B , R  g
of 21cm and a 9 cm air gap (cross-section RA PA P0 A
diameter=4cm). The core was wound 158 turns with
copper wire (cross-section diameter =2mm) to where R is total magnetic reluctance of the core and
become a toroidal magnetic coil. The magnetic flux air gap, P and P 0 are the magnetic permeability of the
density (B) circulating in the coil and air gap can be core and air respectively ( P ~5000 P 0 ), A is the
theoretically expressed in the following equations: cross-section area of the toroidal iron core, N is the

ʳ ˈ˃
ˠ˴̅̆˿˴́˷ʳˣ̅˸̆̆ʳ
˝̂̈̅́˴˿ʳ̂˹ʳ˔̀˸̅˼˶˴́ʳ˦˶˼˸́˶˸ʳ˅˃˃ˌˍʳˈʻ˄ʼʿʳˇˌˀˈˇʳ
˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲ʳ
number of turns of coil, i is the current flowing group one. In order to get more experimental data for
through the coil, lc and lg are the core circumference statistics, we used 50 mung beans of each group
and air gap distance, respectively. The exposure which were put on culture plate will take larger area.
system is shown in Figure 2. We used 60Hz 110Vrms Because the magnetic field intensity decay when
AC electric power as the source and a 60-Watts distance increase, we measured the magnetic field
incandescent lamp as the load, and covered on lamp intensity of the relative position of each group on
with an iron bucket to hide the light of lamp, to avoid culture plate is shown in Figure 2. We got more
other interference for mung beans growth. accurate data were the magnetic field intensity of
We measured the highest magnetic field exposed group one is 875±75mG, exposed group two
intensity of the air gap of coil is 950mG. The air gap is 155±55mG, control group is 1.8±0.8mG.
of coil was to be placed the mung beans of exposed

Fig 2: The exposure system of this experiment

The MF source came from the toroidal magnetic coil that was driven by the 60Hz 110V AC electric power.
To measure the waveform and spectrum of the ELF MF, we used a little probe coil of diameter 3cm (Misakian,
1993) to induce an electromagnetic force close to the coil . The probe was connected to an
oscilloscope/frequency analyzer (Tektronix TDS2012B, Bandwidth:100MHz) to obtain the components of 60Hz
110V AC electric power magnetic field in time and frequency domain are shown in Figure 3 a and b. We found
the waveform of 60Hz 110V AC electric power is distortion and the frequency spectrum with harmonics.
2.3 Methods
We prepared three cylindrical containers with diameters of 5cm and poured into 50ml distilled water, then
put three groups of mung beans in the cylindrical container, respectively. We moved three cylindrical containers
in the positions of rectangular culture plate be shown in Figure 2. After the three groups of mung beans have
been imbibing water for 8 hours, so dehydrated beans were simply rehydrated to allow enzyme reactivation, they
ʳ 51
The Influences of Extremely Low Frequency AC Magnetic Fields At 60Hz on Mung Beans Growth
Pai-Tsun Tien et al.
_________________________________________________________________________________________________________________
were taken out. The three cylindrical containers were removed and three groups of beans were put back in their
original positions of culture plate to continue growing, and then were sprayed into appropriate distilled water by
a sprinkler every 12 hours. Because three groups of mung beans grew on culture plate together, so the
environmental parameters of three groups were almost the same. After mung beans have been growing for 5 days
are shown in Figure 4, three groups of mung bean sprouts were taken out, in general mung bean sprout have two
leaves, and the stem length and leaves length of each mung bean sprout was measured.

(a) (b)
Fig 3: (a) The waveform of 60Hz 100V AC electric power magnetic
field is distorted sine wav˸. (b) The frequency spectrum of 60Hz 100V AC electric
power have harmonics, the measured bandwidth of analyzer is 500Hz.
3. RESULT (P<0.01,one-tailed paired sample t-test). The average
We observed the growth of two exposed groups was leaf lengths of mung bean sprouts exposed to
faster than the growth of control group during 5 days. 875±75mG and 155±55mG ELF MF were great than
The average stem lengths and average leaf lengths of those of control mung bean sprouts (P<0.01,
each group mung bean sprouts were recorded are one-tailed paired sample t-test), too. We can find an
shown in Figure 5. We analyzed experimental data by enhancing effect on the growth of mung bean is
statistical method are shown in Table 1. The average exposed under 875±75mG and 155±55mG ELF MF.
stem lengths of mung bean sprouts exposed to Otherwise, there is no significant different between
875±75mG and 155±55mG ELF MF were great than the exposed group one and exposed group two mung
those of control mung bean sprouts beans (P>>0.05, one-tailed paired sample t-test).

Fig 4: The early growth of exposed 1,exposed 2

and control group mung beans after five days

ʳ ˈ˅
ˠ˴̅̆˿˴́˷ʳˣ̅˸̆̆ʳ
˝̂̈̅́˴˿ʳ̂˹ʳ˔̀˸̅˼˶˴́ʳ˦˶˼˸́˶˸ʳ˅˃˃ˌˍʳˈʻ˄ʼʿʳˇˌˀˈˇʳ
˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲˲ʳ

Fig 5 a: The average stem lengths of each group mung beans. b: The average lengths of first leaf of each group
mung beans. c: The average lengths of second leaf of each group mung beans.

4. DISCUSSION (Smith,1993;Davies,1993; Soja,2003 ;Huang,2007).

According to the reference levels which were So, the growth of plant would be modified when
announced by ICNIRP in 1998 is 833mG (f=60Hz) plant exposed ELF MF intensity above 100 mG for a
for general public exposure to time-varying electric long time. The enhancing influence is abnormal
and magnetic fields, to prevent the influence that may phenomenon for growth of plant, because the motion

cause to the nervous function of human. However, of Ca ion on the cells of plant is changed
the experiment results show that the magnetic field (Lednev,1991;Smith,1993). Therefore, we worry
intensity is 875±75mG and 155±55mG have an about body health would be influenced when human
enhancing effect on the growth of mung beans exposed ELF MF intensity above 100 mG for a long

ʳ 53
The Influences of Extremely Low Frequency AC Magnetic Fields At 60Hz on Mung Beans Growth
Pai-Tsun Tien et al.
_________________________________________________________________________________________________________________
time. We can get magnetic field intensity greater than Effectsof a 60 Hz Magnetic Field on
100mG (rms value), when to measure home electrical Photosynthetic CO2 Uptake and Early Growth of
appliances closely (5cm to 10cm away). To use home Radish Seedlings, Bioelectromagnetics 25(2004):
electrical appliances closely then we would expose 572-581
higher magnetic field intensity, maybe influence the 7. Effects of Weak 162/3Hz Magnetic Fields on
health of human body. So should avoid exposing ELF Growth Parameters of Young Sunflower and
MF intensity above 100 mG for a long time in order Wheat Seedlings, Fischer G, Tausz M, Kock
to reduce the biological effect of extremely low M,Grill D, Bioelectromagnetics
frequency magnetic fields. For electrical appliances 25(2004):638–641
and high-voltage line can induce higher magnetic 8. ICNIRP Guidelines, Guidelines for limiting
field, we should keep the appropriate distance to exposure to time-varying electric, magnetic, and
protect the health of human body. electromagnetic fields (up to 300 GHz) .
(1998):17-18
Corresponding to: 9. IEEE Standard for Safety Levels with Respect to
Pai-Tsun Tien Human Exposure to Electromagnetic Fields, 0–3
Department of Electronic Engineering kHz C95.6, IEEE International Committee on
National Taiwan University of Science and Electromagnetic Safety on Non-Ionizing
Technology Radiation ,(2002):10-15
43 Keelung Road, Da-an District 10. Misakian M. 1993. ELF electric and magnetic
Taipei 106, R.O.C. field measurement methods. 1993 IEEE
Telephone: (886) 2-23443021 International Symposium on Electromagnetic
Fax: (886) 2-23955184 Compatibility. Dallas, TX, Aug 9–13:
E-mail: tyanbt@cht.com.tw Symposium Record. pp. 150–155.
11. Huang HH,Wang SR. 2007. The effects of 60 Hz
REFERENCES magnetic fields on plant growth, Nature and
1. Lednev VV. Possible mechanism for the Science 5(1):60–68.
influence of weak magnetic fields on biological 12. Huang HH,Wang SR. 2008 The Effects of
Systems, Bioelectromagnetics 12(1991): 71-75. Inverter Magnetic Fields on Early Seed
2. Liboff AR. The cyclotron resonance hypothesis: Germination of Mung Beans, Bioelectromagnetic
Experimental evidence and theoretical early view 2008
constraints. In: Norden B. Ramel C. editors.
Interaction mechanisms of low- level
electromagnetic fields in living systems, New
York. Oxford University Press (1992) :130-147.
3. Smith SD, McLeos BR, Liboff AR. effects of
CR-tuned 60Hz magnetic fields on sprouting and
early growth of Raphanus sativus,
Bioelectrochem Bioenerg 32(1993): 67-76.
4. Davies MS. Effects of 60Hz electromagnetic
fields on early growth in three plant species and
a replication of previous results,
Bioelectromagnetics 17(1996): 154-161.
5. Soja G, Kunsch B, Gerzabek M, Reichenauer T,.
Soja AM, Rippar G.,Bolhàr-Nordenkampf HR.
Growth and yield of winter wheat (Triticum
aestivum L.) and corn (Zea mays L.) near a high
voltage transmission line, Bioelectromagnetics
24(2003):91-102
6. Yano A., Ohashi Y,. Hirasaki T and Fujiwara K

ʳ ˈˇ
 Marsland Press
Journal of American Science 2009: 5(1), 55-57
________________________________________________________________________________________________
Letter
The Inflation Dynamics of the ASEAN-4: A Case Study of the Phillips Curve
Relationship
Klarizze Anne M. Puzon
University of the Philippines, Quezon City, Philippines
Email: kmpuzon@gmail.com
____________________________________________________________________________________________________
ABSTRACT: The conventional Phillips curve argues that there is a trade-off or negative relationship between unemployment
and inflation. The aim of this study is to investigate the validity of the Phillips curve for the ASEAN-4 countries: Philippines,
Thailand, Indonesia, and Malaysia from 1980 to 2005. Besides unemployment, the relationship of interest rate, exchange rate,
and supply shocks to inflation, were also investigated. Using various econometric techniques like Ordinary Least Squares and
Instrumental Variables, it was found out that for the ASEAN-4, there seems to be no stable one-to-one trade-off between
unemployment and inflation. Variables that could help control inflation were also different for the four countries. For
Thailand, the inflation lag, unemployment and oil dummy were significant. As for Indonesia, the interest rate, 1997 East
Asian Financial Crisis dummy, and oil dummy were significant in affecting inflation. The OLS regression gave the best linear
unbiased estimate for both countries. For the Philippines, serial correlation was detected. Thus, Prais-Winsten method was
employed. It was then shown that the unemployment lag, interest rate, and exchange rate lag were significant at the 10% level
of significance. [Journal of American Science 2009: 5(1), 55-57](ISSN: 1545-1003)

Key words: Phillips curve; inflation; unemployment; ASEAN

1. INTRODUCTION variables: interest rate, lagged exchange rate, 1997 East Asian
The empirical studies on the Phillips curve analyzing the financial crisis binary dummy, and oil shock dummy variable for
relationship of unemployment rate to the inflation rate are the oil price fluctuations.
results of the search for a tool for forecasting inflation and 
implementing monetary policy. The conventional Phillips curve SW EWEunempt ESWEunempt-1Eintratet + Exrt-1
argues that there is a trade-off or negative relationship between +GGoil+ XW  
unemployment and inflation (Dornbusch, et al. 2005). Economists
soon modified the Phillips curve theory to focus on inflation in The following are the hypotheses for the signs of the explanatory
relation to unemployment. The aim of this paper is to investigate variables:
the validity of the Phillips curve for the ASEAN-4 countries: x Unemployment, unemp, and unemployment lag, unempt-1, as
Philippines, Thailand, Indonesia, and Malaysia from 1980 to 2005. stated by the Phillips curve, is negatively related to inflation.
Some variables that could affect inflation are also analyzed. Thus, That is, if the demand for labor increases due to an
this paper will explore some tools that could aid in the inflation expansionary monetary expansion, the unemployment rate
targeting strategies of the ASEAN-4 economies. would fall causing wages/ prices to rise. Thus, creating a trade-
off between inflation and unemployment.
x The inflation lag, SWthe assumed expected inflation, is
2. EMPIRICAL MODEL positively related to inflation. I assume this using the adaptive
I used annual Consumer Price Index, exchange rate expectations theory.
(domestic currency per dollar), and money market interest rate x The interest rate, intrate or mmintrate, is positively correlated to
data sets supplied by the United Nations Statistical Database inflation. Increasing interest rates results to higher costs for
(UNSD). For each country, the inflation rate was computed as the businesses, which causes prices to rise.
percentage change in the Consumer Price Index. That is, inflation x Due to policy lags, the current exchange rate may be
rate= (CPIt – CPIt-1)/ CPIt-1 * 100. All CPI and inflation rates data endogenous. Thus, I assume that the exchange rate lag is
would have 2000 as the base year (CPI= 100). In addition, since exogenous and use it in the model. The exchange rate I use is in
the UNSD only have survey data for unemployment, we acquired the form: domestic currency per dollar. I use xr_1 to account
for trade prices. I hypothesize that an increase in xr_1, a
more reliable unemployment rates from the National Economic
depreciation of the local currency, would increase inflation
Development Authority of the Philippines website. All of the because of a higher import prices.
annual data sets covered the period from 1977 to 2005. x Binary dummies, 97 and oil, were added to account for price
For the empirical model, I modified the equations by shocks brought by the 1997 financial crisis and oil crises. Such
Gordon (1997), Dua (2006), Stiglitz (1997), Staiger, Stock, and control variables are expected to have a positive sign because
Watson (1997), and Smith (2000). In the formulation of a simple they serve as supply shocks. To account for East Asian
augmented Phillips curve, I also utilized Wan’s (2001) linear financial crisis, the years 1997 and 1998 have their 97 dummy
model involving lagged inflation and cyclical unemployment as equal to one. Meanwhile, the oil dummy for 1980, 1990, and
explanatory variables. 2005 is equal to unity since oil price fluctuations occurred
For Equation 1, I use the augmented version of Stiglitz’s during those years.
model to capture inflationary expectations by including the lagged
inflation rate as a measure of the expected inflation rate. In For Equation 2, I use first differencing. This model will only
addition, I include an unemployment lag to determine if such be used if the equation experiences unit root problems. Such unit
would provide a better fit. I also have additional explanatory root behavior was tested using the Phillips-Perron test.

55
The Inflation Dynamics of the ASEAN-4: A Case Study of the Phillips Curve Relationship
Klarizze Anne M. Puzon
___________________________________________________________________________________________________
 becomes more expensive. For Indonesia, the significant variables
'SW D'Eunempt 'ESW'Eunempt-1'Eintratet + at the 10% significance level were the interest rate, oil and 1997
'Exrt-1 +GGoil+ XW   financial crisis dummies. The dummy for the 1997 East Asian
 financial crisis, surprisingly, depresses predicted inflation by about
To have more efficient estimates, I tested Equations 1 or 14.92 percentage points. More specifically, ceteris paribus, during
2 for heteroskedasticity and serial correlation. If either problem the 1997 East Asian financial crisis, inflation decreases by
exists, corrections are employed to ensure consistent estimates. As approximately .581- 14.92= |-14.339| percentage points. Such
will be discussed later, I also used Instrumental Variable method result, most probably, was due to lower productivity growth and
for Malaysia. More specifically, since unit root behavior occurs in aggregate demand in the economy. This finding was similar to that
the inflation variable, I used an instrument, the inflation lag of of Vong (2001) in his study of Macau’s Phillips curve. Meanwhile,
Singapore, for the inflation lag of Malaysia. the oil dummy indicated that, in the presence of oil price shocks,
ceteris paribus, predicted inflation for Indonesia, is 4.9076
3. Discussion of Regression Results percentage points higher than usual. That is, when there are oil
Using t-test, with an Ho: Bj=0, and a two-sided price shocks, inflation increases by .581 + 4.9076= 5.4886
alternative of H1: Bj  0, the results for Equation 1 can be percentage points. In addition to these, it was seen that
summarized as follows: unemployment and its lag were not statistically significant. The
unemployment variables also had positive signs which could
Table 1. Fully-corrected regression results for Equation 1 indicate that there maybe no trade-off between inflation and
Dependent variable: Inflation unemployment. Being a developing country, it seems to be that
Significance level: 10% Indonesia suffers from both persistent high inflation and high
Explanatory Variable Indonesia Malaysia Thailand Philippines unemployment rates.
Infla_1 -.0488733 .3483649 .3773198 -.0074889 For the Philippines, using OLS, it was found out that the
(.1096472) (.2066877) (.1637375) (.1133733) unemployment lag, interest rate, and exchange rate lag were
Unemp 0.2920096 -.4904054 -.9443959 1.164838 significant at the 10% level. For a one percentage point increase in
(.7119024 ) (.5649236) (.4813811) (1.157631) the unemployment lag, inflation decreases by 2.0367 percentage
Unemp_1 .0188285 .3516186 -.389395 -2.008291
(.8245843) (.5329042) (.5979983) (.7032352) points. Such supports the trade-off between unemployment and
Mmintrate 1.200941 -.060483 -.3699473 2.594078 inflation as indicated by the Phillips curve. That is, if the demand
(.1402781) (.1525026) (.3366451) (.2925591) for labor increased due to an expansionary monetary policy, the
Xr_1 .0001955 -1.265882 -.1813196 .8965375 unemployment rate would fall. Then, wages and consumer prices
(.0008092) (1.38491) (.1655893) (.3458095)
D97 -14.92133 1.656048 4.670027 -3.558471 will tend to rise. Moreover, the significance of the unemployment
(5.921631) (1.073852) (2.812547) (3.915062) lag could indicate that fiscal policies relating to inflation might not
Doil 4.907661 1.596374 5.304789 4.543862 have an immediate effect. There could be policy lags. Meanwhile,
(2.690442) (1.078417) (1.658721) (4.269187) a percentage point increase in interest rates increases inflation by
Constant .5815264 .07467 -.9713239 -.2621564
(.9750957) (.830654) (.566889) (.9492679) 2.613 percentage points. In addition, when the exchange rate lag
R2 0.8855 0.4730 0.6918 0.8244 increases by one percentage point, inflation increases by 0.8893
Adj R2 0.8409 0.2680 0.5720 0.7561 percentage points. This supports our hypothesis that depreciation
n 26 26 26 26 in the domestic currency makes local goods more competitive.
Such increases aggregate supply and results to an increase in the
For Thailand and Indonesia, the OLS regression gave price level. However, even though the Philippines’ OLS model
the best linear unbiased estimate (BLUE). For both countries, the gave significant results, it is not BLUE. Using Durbin’s alternative
error terms have constant variance and have no autoregressive test for autocorrelation, with a p-value of 0.0167, at the 10%
conditional heteroskedasticity (ARCH). There were also no significance level, there was evidence that the Philippines’
random walk and serial correlation problems. For Thailand, using Equation 1 regression suffers from serial correlation. Generally,
the adjusted R2 value, 57.20% of the inflation variation was when corrected for serial correlation, I have seen that the standard
explained by the model. This is an improvement compared to errors decreased. Although they are characterized by lower
Equation 1’s adjusted R2 value of 23.44%. Moreover, the inflation coefficients, unemployment lag, interest rate, and exchange rate
lag, current unemployment rate, and oil dummy were significant at remain significant. For a one percentage point increase in the
the 10% level. The significance of the inflation lag is consistent to unemployment lag, inflation decreases by 2.0082 percentage
the findings of Dua (2006). This may signal that inflation is points. On the other hand, a percentage point increase in interest
inertial (Smith, 2000). For Thailand, past inflation influences rates increases inflation by 2.594 percentage points. In addition, a
people’s expectations on future inflation. Meanwhile, it was also one point percentage increase in the exchange rate lag increases
estimated that a one percentage increase in unemployment, inflation by 0.8965 percentage points.
decreases inflation by .94439 percentage points. Such value is For Malaysia, the OLS model explains 43.15% of the
very near to one. Thus, I could say that the Phillips curve is variation in inflation. Only unemployment was significant at the
present in Thailand using 1980-2005 data. Unemployment and 10% level. A one percentage point increase in unemployment
inflation have a trade-off. This finding is similar to that found by decreases inflation by 1.543 percentage point. Such finding is still
Bhanthumnavin (2002). Oil price shocks also influenced Thai consistent with the OLS estimates of Tang and Lean (2007): that
inflation values immensely. It was implied that, controlling for all there exists a trade-off between unemployment and inflation in
other variables, when there is an oil price shock, predicted Malaysia. However, while this might support the Phillips curve
inflation is about 5.30 points higher than for a year without an oil hypothesis, we should be careful with the interpretation of results.
price shock (i.e. inflation= doil + constant). That is, when there are This is because, when tested for unit root behavior using the
oil price fluctuations, inflation increases by about 5.30 + (-.917)= Phillips-Perron test, with a p-value of 0.1298, it was found out that
4.329 percentage points. From these, I could see that as the price the past values of inflation were correlated. In addition, the
of energy rises, the inflation rate will increase as production inflation lag might be endogenous. It might be correlated with the

56
Marsland Press
Journal of American Science 2009: 5(1), 55-57
________________________________________________________________________________________________
error term. To solve for this problem, I use the inflation lag of I will now focus our discussion on the theorized Phillips
Singapore as an instrumental variable for Malaysia’s inflation lag. curve relationship: trade-off between unemployment and inflation.
I use Singapore data since I thought that its price levels might be I will analyze the signs of the unemployment rate for the four
highly correlated with that of Malaysia. Such may be a result of countries without emphasizing their significance at the 10% level.
their geographical proximity and trading relations. The simple Using the fully-corrected models, it was found out that for
correlation of Malaysia’s inflation lag with Singapore’s inflation Thailand and Malaysia, there exist a trade-off between
lag was 0.7162. In addition, when Malaysian inflation lag was unemployment and inflation. The negative coefficients for
regressed with all other exogenous variables and the Singaporean unemployment are the evidences for this. The trade-off is
inflation lag, it was found out that Singapore’s inflation lag, with a approximately one-to-one for the two countries. Such relationship
p-value of 0.079, was significant. This supports one of the supports the Keynesian view on the Phillips curve. That is, at least
assumptions for an instrument. The covariance of our instrument, for the short-run, unemployment and inflation have a negative
Singapore’s inflation lag, and our xj, Malaysia’s inflation lag, is relationship. In contrast, the unemployment coefficients for
not zero. Meanwhile, I assume that Cov(Singapore_inflation_1, Indonesia and the Philippines were positive. The findings for the
u)= 0. When I used Singapore_inflation_1 as an instrument for Philippines are consistent with Dua’s findings (2007). The
infla_1 in our Malaysian OLS model, the inflation lag and positive relationship between unemployment and inflation is
unemployment were significant at the 10% level. The inflation lag supported by Rational Expectations Theory. There may be no
fulfilled our expected sign. However, again, we could not be sure trade-off between unemployment and inflation because markets
as to the reliability of these results. Using the Phillip-Perron test, respond quickly to changes in prices and wages.
there was an evidence of a highly persistent time series. The past
values of inflation are still correlated. Thus, I use Equation 2, the 4. Conclusions and Recommendations
first-differenced model, for our analysis. The regression with For the ASEAN-4, significant or not, there seems to be
Equation 2 showed that there seems to be no significant variables no stable one-to-one trade-off between unemployment and
which could affect inflation. Such results might be consistent but inflation. I also found out that the variables which could help
not efficient. This is because of the presence of large standard control inflation were different for the four countries. Meanwhile,
errors caused by either heteroskedasticity or serial correlation. to have more conclusive results and achieve normality, I suggest
When tested for both stationary and autoregressive conditional obtaining a bigger sample size, e.g. usage of quarterly data. In
heteroskedasticity, the first-differenced model was characterized conducting tests, such would give us higher degrees of freedom. In
by homoskedasticity. However, when tested for serial correlation addition, for serial correlation problems, error terms such CPI
of order 1, AR(1), and higher order correlation using the Breusch- minus unit labor cost can be used (Smith, 2000). I also suggest a
Godfrey LM test for autocorrelation, it was evident that the lag for interest rates. There maybe a possibility that the previous
Equation 2 for Malaysia suffers from serial correlation. With these, year’s monetary policy regarding interest rates might have a
we have seen that although differencing could eliminate most of significant effect on the inflation rate. In addition, to better explain
the serial correlation, it has not done so for our model. Most inflation dynamics, stock prices, energy/ petroleum prices, and
probably, our model suffers from higher order serial correlation. other functional forms (e.g. quadratic or logarithmic form) can be
To correct for serial correlation, I use Prais-Winsten estimation. utilized in future studies. Furthermore, panel data analysis could
When corrected for serial correlation, the first-differenced be utilized. Lastly, cointegration tests may be employed to explore
equation, Equation 2, had lower standard errors. This shows that the feedback dynamics of employment- inflation relationship.
the existence of serial correlation produced large standard errors.
Correspondence to:
For the fully-corrected model, it was only the inflation lag, with a Klarizze Puzon
p-value of 0.109, which is nearly significant at the 10% level. Email: kmpuzon@gmail.com
From the regression results, it can be seen that as the instrumented
inflation lag increases by one percentage point, inflation increases REFERENCES
[1] Bhanthumnavin, K. (2002). The Phillips curve in Thailand. St. Antony’s College,
by .3483 percentage points. The nearly significant value might University of Oxford Working Paper.
have been the result higher order autocorrelation. The model [2] Dornbusch, R., Fisher, S. & Startz, R. (2005). Macroeconomics, Ninth Edition.
might not have been fully-corrected because I also used Prais- U.S.A.: McGraw-Hill Education.
[3] Dua, P. (2006). Inflation- Unemployment Trade-off in Asia. United Nation’s
Winsten method—a method which only employs feasible GLS Project LINK Meeting. University of Delhi.
estimation of AR(1). In addition, we should also take note that the [4]Gordon, R. (1997). The Time-Varying NAIRU and Its Implications for Economic
FGLS is not unbiased and therefore, is not BLUE. Moreover, Policy. Journal of Economic Perspectives, American Economic Association, vol.
although it may be asymptotically more efficient than the OLS 11(1), pages 11-32, Winter.
[5] National Economic and Development Authority, Republic of the Philippines.
estimator in the presence of serial correlation, we cannot fully (2007). http://www.neda.gov.ph/.
assume weak dependence because of a small sample size of 26. [6] Smith, H. (2000). The Phillips Curve in the 1990s. University Avenue
Another possible reason for our findings is that Undergraduate Journal of Economics, Year 2000 Issue.
[7] Staiger, Douglas, James Stock, and Mark Watson. (1997). The NAIRU,
Singapore_inflation_1 might not be a completely exogenous Unemployment and Monetary Policy. Journal of Economic Perspectives (Winter) pp.
instrument for Malaysia’s inflation lag. Our IV, Singapore’s 33-49.
inflation lag might be correlated with the error term. This could [8]Stiglitz, J. (1997). Reflections on the Natural Rate Hypothesis. Journal of
happen because Malaysia and Singapore are closely-linked Economic Perspectives,Winter, Vol. 11(1), 3-10.
[9]United Nations. United Nations Statistics Division. (2007).
economies. For example, there is a possibility that the exchange http://unstats.un.org/unsd/default.htm.
rate between the currencies of the two countries is correlated to [10] Vong, T. (2001). The Phillips Curve in Macau and Its Implications for
our IV. Thus, Singapore’s inflation lag might not be the best IV Macroeconomic Policy. Monetary Authority of Macau Publication.
[11] Wan, Y. (2001). A Case of the Phillips Curve in the Formation of a Monetary
for Malaysia’s inflation lag. With this, it is recommended, that in Union: A Glimpse at High Inflation Countries of the Monetary Union. University
future studies, the exchange rate lag, the interest rate lag, and Avenue Undergraduate Journal of Economics, Year 2001 Issue.
others be tested as possible instrumental variables.

57
Marsland Press
Journal of American Science 2009: 5(1), 58-69
______________________________________________________________________________________
Research Article
Estimating Soil Loss Using Universal Soil Loss Equation (USLE) for
Soil Conservation planning at Medego Watershed, Northern Ethiopia
Gebreyesus Brhane and Kirubel Mekonen
Center for Development Research (ZEF), University of Bonn, Walter-Flex-Str. 3,
D-53113 Bonn, Germany
E-mail: gebre042001@yahoo.com
ABSTRACT: Water erosion is a major part of land degradation that affects the physical and
chemical properties of soils and resulting in on-site nutrient loss and off-site sedimentation of water
resources in arid and semi-arid areas of Ethiopia. The heavy reliance of some 85 percent of
Ethiopia’s growing population on an exploitative kind of subsistence agriculture is a major reason
behind the current state of land and soil degradation. Tackling on-site effects of soil erosion requires
understanding of the rates of soil loss as well as identification of the major controlling factors that
enhance or retard these processes. Therefore, the objective of this study was to predict the amount of
soil loss in different landforms and land uses using USLE which is modified and adapted to
Ethiopian conditions, at Medego watershed, northern Ethiopia. This study was conducted after
massive SWC practices have been implemented in the past 15-year in the study watershed. Primary
data and secondary data were collected related to the factors that influence soil loss estimated by
USLE and for area description. The land surfaces in the watershed is mainly a reflection of past
erosion processes as indicated by many researchers. In this study, the lowest soil loss is estimated on
flat plains (< 2% slope) about 1.59 tons ha-1 y-1, which is less than the minimum tolerable soil loss (2
tons ha-1 y-1) for the country. However, the highest soil loss is from steep slopes (30-50%) which is
35.43 tons ha-1 y-1, about twice the maximum tolerable soil loss (18 tons ha-1 y-1). The average soil
loss rate at watershed level is 9.63 tons ha-1 y-1 about half of the maximum tolerable soil loss. The
implication is the contribution of the implemented SWC measures in decreasing the rate of soil
erosion is encourageable as compared to the results related to high soil loss estimated in the past
studies i.e., before massive SWC implementation. However, the present value indicates still a need
for wise SWC planning that decreases the amount of soil loss in the watershed at least below the
maximum tolerable soil loss rate of the country. Therefore, to maximize the available resources in
targeting the effect of water erosion on soil loss, those landforms and land uses having large rate of
erosion should be given first priority during the introduction of intensive and well designed SWC
interventions at Medego watershed, northern Ethiopia. [Journal of American Science 2009: 5(1), 58-
69] (ISSN: 1545-1003)

Key words: Medego watershed, northern Ethiopia, soil loss, tolerable soil loss.
Abbreviations: SWC- soil and water conservation, USLE-Universal Soil Loss Equation.

1. INTRODUCTION lost through erosion in a watershed actually makes

Growing degradation and loss of soil means that it to the ocean as sediment (FAO/UNEP, 1978).
the expanding population in many parts of the The remaining three-fourths are deposited on
world is pushing this resource to its frontier. In its foothill slopes, in reservoirs, in river plains and
absence, the biospheric environment of humans other low-lying areas or in the river-bed itself,
would collapse with devastating effects on which often causes channel shifts. In an overview
humanity. Judson (1965) was one of the first of global erosion and sedimentation, Pimental
geologists to assess the world soil erosion. He (1995) stated that more than 50% of the world’s
estimated that the amount of river-borne soil pastureland and about 80% of agricultural land
carried into the oceans had increased from 9.9 suffer from significant erosion.
billion tons a year before the introduction of
agriculture, grazing and related activities, to the The causes of land degradation are
present rate of 26.5 billion tons a year. complex and have diverse nature and dimensions,
Hydrologists estimated that one-fourth of the soil depending on peculiarities of different countries,

58
Estimating Soil Loss Using Universal Soil Loss Equation (USLE) for Soil Conservation planning at Medego Watershed, Northern
Ethiopia
Gebreyesus Brhane et al.
______________________________________________________________________________________
influenced as it is by a combination of natural and by FAO (1986), some 50% of the highlands of
socio-economic-cultural factors. In Ethiopia, the Ethiopia are already ‘significantly eroded,’ and
heavy reliance of some 85 percent of Ethiopia’s erosion causes a decline in land productivity at the
growing population on an exploitative kind of rate of 2.2% per year. The study also predicted that
subsistence agriculture is a major reason behind by the year 2010, erosion could reduce per capita
the current state of land degradation. Moreover, incomes of the highland population by 30%.
land degradation is a long-term process in which Hence, soil and water conservation measures have
the effect and steady expansion is hardly noticed been implemented to alleviate both problems of
until it manifests itself with disastrous drought and erosion and drought, which are symptoms of two
famine. Most studies indicate that sheet and rill different extremes of rainfall conditions since the
erosion by water and burning of dung and crop 1980s in the country. However, so far, little or no
residue are the major components of land sufficient documented information has been
degradation that affects on-site land productivity available on the contribution of the different SWC
(Hurni, 1993; Zeleke et al., 2001). measures implemented on soil loss reduction since
the last 15-years at the study watershed in the
Water erosion is a major part of land semiarid areas of Ethiopia as compared to the
degradation that affects the physical and chemical tolerable soil loss determined by Hurni (1985) to
properties of soils and resulting in on-site nutrient Ethiopia condition. Such information is vital to
loss and off-site sedimentation of water resources take additional measures and soil conservation
in arid and semi-arid areas like Ethiopia planning at the watershed and other similar areas
(Boardman, 1998; Lal, 1999; Bartsch et al., 2002; in the semiarid areas. Therefore, the purpose of
Emrah et al., 2007). The off-site effects of erosion this study was to estimate the amount of soil loss
such as reservoir sedimentation and water in different landforms and land uses using USLE
resources pollution are usually more costly and at Medego watershed, northern Ethiopia.
severe than the on-site effects on land resources
(Phillips, 1989). Therefore, proper management of
on-site effect of soil erosion could reduce the risks 2. MATERIALS AND METHODS
and negative impacts of down stream water
resources due to water erosion. Tackling the on- 2.1. Study Area Description
site effects of soil erosion requires an
understanding of the rates of erosion processes as The study was carried out at Medego watershed in
well as identification of the major controlling the administrative unit of Lalay-maychew district
factors that enhance or retard these processes. The in Tigray region, northern Ethiopia (Figure 1),
knowledge of “what are the factors and where” from August 2007 to July 2008. Its altitude ranges
may help to distinguish the potential causes and from 2000 to 2720 m above sea level. The study
the associated reasons behind the respective causes area is bounded by latitudes N14o05.955’and
even though this may not be enough to design site- 14o05.937’, and longitudes E038o42.352’ and
specific management, as the factors playing a 038o42.333’. The total area of the watershed is
major role in erosion may be widely distributed about 1091.5 ha as delineated using Geographical
within watersheds (Ferro et al., 1998; Mirco et al., Positioning System (GPS) during the field study.
2003). The study watershed is characterized by different
landforms which are ranged from flat plains,
Soil erosion by water and its associated undulating plains and rolling land to steep
effects are recognized to be severe threats to the mountains and very steep escarpments. The
national economy of Ethiopia (Hurni, 1993; description of the topography is adopted the slope
Sutcliffe, 1993, Tamene, 2005). Since more than capability classification made by (Chekun, 2002),
85% of the country’s population depends on and the slope ranges and area coverage of each
agriculture for living, physical soil and nutrient landform was recorded at field using clinometers
losses lead to food insecurity. Hurni (1990, 1993) and GPS, as it is presented in Table 1. The
estimates that soil loss due to erosion in Ethiopia geological setup of the watershed is originated
amounts to 1493 million tons per year, of which from volcanic. However, alluvial deposits at flat
about 42 tons ha-1 y-1 is estimated to have come lands are also found in the watershed. The soil
from cultivated fields. This is far greater than the type at the study watershed is quite different along
tolerable soil loss as well as the annual rate of soil the slope. At steep slopes, coarse earth materials,
formation in the country. According to an estimate gravels and boulder are dominated where as at flat

59
Marsland Press
Journal of American Science 2009: 5(1), 58-69
______________________________________________________________________________________
plain, the largest portion of the study area is comoldulensis) and some ‘seraw (Acacia etbaica)
covered by clay loam soil and the smaller portion on cultivated and marginalized areas. Leucena
laid on clay and sandy soil textures. The main soil (Leuceana leucacephala), sesbania (Sesbania
types are cambisols on undulating plains and sesban) and some other grasses are commonly
rolling landforms; lithosols on hilly and steep to found in the gully of the watershed. Farmers' used
very steep lands and vertisols are found on the flat such vegetation for the purpose of farm implements,
plateau plains of the watershed (BoARD, 2007). house construction and furniture, fuel wood, soil
and water conservation measures and fencing
The number of households and total (Table 2). But most farmers have no or little
population at Medego watershed is 397 and 1537, awareness on the function of these tree species for
respectively. The land holding size of most soil and water conservation as compared to the
farmers in the study area is less than 1.3 ha. The other uses. Hence, awareness creation to farmers in
watershed has uni-modal and erratic rain fall the watershed and other areas should be done in
patterns. The rainy season is very short and order to the farmers give attention on planting and
extends from June to first week of September. The managing tree species from different perspectives
mean annual amount of rain fall ranges from 600 - including soil and water conservation, soil fertility
700 mm from historical rainfall data. The mean improvement.
monthly temperature during the growing season
ranges from 15 - 20 o C (BoARD, 2007). 2.2. Methodology
According to the BoARD (2007), the farming
system of the study watershed is principally crop Primary and secondary data were collected at
oriented. Tef cultivation (Eragrostic tef) account Medego watershed related to the assessment of
for the majority of arable lands and followed by SWC measures on soil loss at Medego watershed.
wheat (Triticum vulgare) crop. Other crops such as Primary data were gathered by topographic transect
faba bean (Vicia faba), field pea (Pisum sativum), walk, measuring of input data, informal discussion
lentil (Lens culinaris), chick pea (Cicer arietinum), and observation. The secondary data include
flax (Linum usitatissimum), barley (Hordeum climate, demographic and other related data were
vulgare) and maize (Zea mays) are also important collected from Bureau of Agriculture and Rural
crops in the farming system. Irrigation is also Development (BoARD) at the administrative unit.
widely practiced at Medego watershed. In spite of These data were used to estimate soil loss after
the fact that the high crop diversification in the tremendous activities of SWC measures have been
watershed, it observed that there is still a room to implemented at Medego watershed, northern
improve the crop productivity. Livestock rearing is Ethiopia. The rapid rural appraisal technique of the
also an integral part of the farming system, though topographic transect walk method was employed
the number of livestock in the watershed area is for its effectiveness in the assessment of the natural
reduced from time to time due to animal feed resource base and topography of the watershed. In
shortage. According to farmers view, cattle are order to obtain as much information as possible,
kept mainly for draught power and milking; goat the transect walk was applied in two direction, east
and sheep are kept for live sale; and equines to west and south to north. In both directions, the
(donkey, mule, horse, camel) for transportation. transect walk started at the top edge of the
The study indicated that 83% of the households in watershed and went all the way across to the other
the watershed have some livestock. Of these, 75% end of the watershed. During the transect walk,
are cattle (average of 2 cattle per household), 21% observations and estimates of vegetation type and
are sheep and goats and the rest is covered by density, and impact of the existing soil and water
poultry and equines. conservation measures were observed. These were
followed by recording land-use types, soil color,
It was observed that the vegetation in soil depth, soil drainage condition, slope gradient
Medego watershed in general is sparse and has and length. The transect walk also provided an
been overexploited for long time and at this time opportunity for informal discussions with farmers
consists of shrubs and bushes of little economic working on their plots.
value. The available vegetation species in the study
area include seraw (Acacia etbaica), chea’ (Acacia Annual soil loss in the form of runoff
abyssinica), acacha (Acacia decurrence) and Awhi from different land forms and land uses of the
(Cordia africana) on uncultivated land; and watershed was estimated using the Universal Soil
momona (Acacia albida), tambock (Croton Loss Equation (USLE) (Wischmeier and Smith,
machostachys), keyih bahrizaf (eucalyptus

60
Estimating Soil Loss Using Universal Soil Loss Equation (USLE) for Soil Conservation planning at Medego Watershed, Northern
Ethiopia
Gebreyesus Brhane et al.
______________________________________________________________________________________
1978) and modified and adapted to Ethiopian landform and land use and so extrapolated based
conditions by Hurni (1985) and Gebreselassie on Hurni (1985) to Ethiopia condition (Table 3 and
(1996) as follows. 4).

A=R*K*L*S*C*P The C-factor is defined as the ratio of soil

loss from land with specific vegetation to the
Where; A = estimated soil loss (t ha-1 yr-1), R = corresponding soil loss from continuous fallow
Rainfall Erosivity factor, K = Soil Erodibility (Wischmeier and Smith, 1978). Assessment of the
factor, L = Slope length factor, S = Slope gradient type of land use-cover was made separately for
factor, C = Land cover factor, P = Management each land unit and the corresponding land cover
practice factor was obtained from Hurni (1985) which was
developed to Ethiopia condition. For variations in
The R-factor is defined as the product of land cover with specific land unit or landform, the
kinetic energy and the maximum 30 minute C-factor was obtained using weighted value of the
intensity and shows the erosivity of rainfall events different land cover (Table 3).
(Wischmeier and Smith, 1978). However, in this
study, to determine the value of the R-factor, the The P-factor gives the ratio between the
average of annual historic rainfall event (10-years) soil loss expected for a certain soil conservation
was collected from meteorological station located practice to that with up-and down-slope ploughing
at 8-Km distance from the watershed. Then the R- (Wischmeier and Smith, 1978). Specific
value corresponds to the mean annual rainfall of cultivation practices affect erosion by modifying
the watershed was found using the R-correlation the flow pattern and direction of runoff and by
established in Hurni (1985) to Ethiopia condition. reducing the amount of runoff (Renard and Foster,
Therefore, the annual R-factor for the average 1983). In areas where there is terracing, runoff
rainfall (650 mm) at the watershed as extrapolated speed could be reduced with increased infiltration,
from Hurni (1985) is 357. The soil erodibility (K), ultimately resulting in lower soil loss and sediment
slope length (L), slope gradient (S), C, and P- delivery. Values for this factor were assigned
factors of USLE for the entire watershed based on considering local management practices and based
landforms and land use is presented in Table 3. on values suggested in Hurni (1985). Management
factors were obtained by assessing the different
The K-factor is defined as the rate of soil supporting practices in the study watershed and it
loss per unit of R-factor on a unit plot (Renard et was taken the weighted value for similar land
al., 1997). To determine the value of the K-factor, forms and land uses types (Table 3). The data
a systematic observation on soil color of watershed related to management practices were collected
was carried out, based on the approach described during the field work. The presence and status of
in Hurni (1985). This was done by classifying the conservation activities were assessed with
watershed into similar land uses and land forms emphasis on the existing conditions of terraces and
(Table 1). For soils having different color in the protected areas. Most of the areas in the watershed
same land use and landform, the K-factor was are well-terraced, mainly the upslope parts.
taken as their mean value of these colors as it is However, most of the terraces are broken due to
described on Hurni (1985). As an example, the K- high runoff and/or livestock trampling in many
factor for flat plains in Medego watershed is the parts of the watershed.
mean value of the soil color black (0.15) and
brown (0.2), which is about 0.18; and the same The data were analyzed following the
approach was used in determining the soil color interpolation of the values of USLE in Hurni (1985)
for the other landforms in the watershed (Table 3). and Gebreselassie (1996) to Ethiopia condition.
SL is the topographic factor expressed as the The data was then interpreted qualitatively and
expected ratio of soil loss per unit area from a field using descriptive statistics.
slope to that from a unit plot under otherwise
identical conditions. Slope length and slope
gradients factors were recorded using meter tape 3. RESULTS AND DISCUSSION
and clinometers, respectively, in the watershed on
different landform and land uses. It is taken the 3.1. Estimated Soil Loss Using USLE at Medego
weighted value of the slope gradient and slope Watershed, Northern Ethiopia
length range measured at the field for each

61
Marsland Press
Journal of American Science 2009: 5(1), 58-69
______________________________________________________________________________________
In spite of the fact that tremendous efforts of SWC same time decrease runoff amount. Moreover,
have been implemented, their contribution in there are rock-out crops, which are difficult to
reducing soil loss due to water erosion demands detach or transport by raindrops and water erosion
recent assessment for appropriate future on the very steep escarpment of Medego watershed.
conservation planning. It is understood that heavy Landforms more than 50% slopes are protected
rainfall cause severe soil erosion in agricultural areas in the watershed. The C-factor represents
fields of the semiarid regions of Ethiopia. Soil resistance of the ground surface to the transport of
erosion in agricultural fields affects not only land water-soil mixture on the very steep mountains of
productivity but also the water environment in the the watershed includes badlands hard, and bushes
down stream. Many investigations have been and shrubs which dissipate the force of the
conducted for the development of prediction raindrops. The P-factor stands for erosion
methods of water-induced soil erosion processes. inhibition effect, and reflects partly awareness and
Among the methods, the empirical Universal Soil control measures implemented to minimize soil
Loss Equation (USLE) has been applied broadly erosion more than the other landforms by the
for predicting the average annual soil loss from community (Table 3). It is also noted that the
upland fields in Ethiopia (Wischmeier and Smith, lower slope landforms are susceptible to daily
1978; Hurni, 1985) for the reasons described in the human interferences where as the steepest
discussion part of this paper. landforms are protected areas. This proves that the
USLE is useful for assessing the adequacy of
The soil loss estimated using USLE on conservation measures and management practices
this study from cultivated land on flat plain land in agricultural watersheds.
form (< 2% slope) of Medego watershed, northern
Ethiopia is the lowest as compared to the other The average annual soil loss estimated by
land uses or landforms, which is 1.59 tons ha-1 y-1 USLE from the entire Medego watershed is 9.63
(Table 4). This indicates that soil loss due to rill tons ha-1. If we interpret the annual soil loss as a
and inter-rill erosion is almost balanced by proxy to watershed erosion, it is possible to see
deposition within the flat landforms of the that the magnitude of annual soil loss reported in
watershed. Next to the flat land form, the Table 4 is generally higher than the tolerable soil
landforms having lower soil erosion are undulating loss of 2 – 18 tons ha-1 y-1 estimated for Ethiopia
plains (slope 2-8%) and flat- flood prone areas (< by (Hurni, 1985) except flat landforms of the
2% slope) which are 3.13 and 4.87 t ha-1 y-1, watershed. The soil loss rate in all the landforms
respectively. The highest soil loss at the study are below the maximum tolerable soil loss for
watershed was recorded at the landform -steep Ethiopia condition, which is 18 tons ha-1 y-1,
-
mountains (slope 30-50%), which is 35.43 tons ha except the steep mountains (slope 30-50%)
1
yr-1. The small soil loss rate of the landforms is landforms that indicate almost double of the
related to the factors of the USLE in the watershed maximum soil tolerance value. In general, the
(Table 4). Therefore, more attention should be average soil loss in the watershed in about half of
given to slope ranges between 30-50% while SWC the maximum tolerable soil loss and five times the
measures is planning to implement in the minimum soil loss tolerance value given by Hurni
watershed. (1985). The implication is that there is a need to
integrate a sound management practices so that to
The general trends of the finding indicate decrease the amount of soil loss in Medego
that soil loss increases as the slope steepness watershed, northern Ethiopia below the maximum
increases in the watershed (Table 4). However, at as well as the minimum soil loss tolerable value
the landform of very steep mountains (> 50% for the country.
slope), the annual soil loss is estimated as 7.63
tons ha-1, which is even less than the landforms As compared to the soil loss estimated for
such as rolling land forms (8-15% slope), hill Ethiopia as 42 tons ha-1 y-1 from cultivated fields
landforms (15-30% slope) and steep mountains by Hurni (1990, 1993); 21 tons ha-1 y-1 (Machado
(30-50% slope). This is because these slope ranges et al. 1995), and 30-80 tons ha-1 y-1 (Tekeste and
are susceptible to daily human interferences such Paul, 1989) in Tigray region, northern Ethiopia,
cultivation and grazing as compared to very steep the soil loss estimated on this study in 2007/08 is
slopes (> 50%) and also slopes having more than by far the smallest. The results of the present study
50% in the watershed have land cover of ‘Bad as compare to past findings indicate that the
Lands Hard’ and stone cover which can retain the amount of soil loss from a given unit of land is low.
impact of the kinetic energy of raindrops and at the This could be due to the contribution of the

62
Estimating Soil Loss Using Universal Soil Loss Equation (USLE) for Soil Conservation planning at Medego Watershed, Northern
Ethiopia
Gebreyesus Brhane et al.
______________________________________________________________________________________
different soil conservation interventions which would occur under alternative management
implemented for at least the last decades in the strategies and practices in order to minimize the
country in general and the study watershed in costs related to fertilizer and environmental
particular. This related to the fact that SWC rehabilitation.
intervention increases soil moisture, fertility and
decrease slope factor and thereby enhance the
availability of vegetation covers. The combined
effect of such factors will be decreasing the impact
of raindrops, detachment and transporting of soils.
This was evidenced by the opinion of the 4. DISCUSSION
respondents which evaluated as less soil erosion
after the soil conservation practices were built at 4.1. Soil Erosion Models and Their Potentials and
the watershed as compared to before the Challenges
implementation (data not presented here).
Therefore, as noted in the above, the soil loss Soil erosion is the most serious causes of land
estimated by different scholars has showed degradation have influenced tremendous pressure
discrepancy for the same environment (semi-arid on productivity and environmental stability of arid
region of Ethiopia). This implies that there is a and semiarid areas. Serious impacts led the
need to have site specific (watershed level) demand for conservation and management
information on soil erosion in order to support measures to reduce the magnitude of soil loss and
timely information for decision makers so that to the extent of its associated impacts in many parts
plan the correct soil conservation planning. In of the arid and semiarid areas. There are many
doing so, it is categorized the severity of erosion in models in existence estimating soil erosion. The
the study watershed’s landforms as follows. USLE has the advantage of being less data
demanding than other models. A wide range of
According to Singh and Phadke (2006) models that differ in their data requirement for
classes of soil loss range (very slight, slight, model calibration, application, complexity and
moderate, severe and very severe), the mean processes considered are available for use in
annual soil loss (9.63 tons ha-1) from Medego predicting soil loss (Merritt et al., 2003).
watershed, northern Ethiopia is categorized under Physically based spatially distributed soil erosion
slight class of soil erosion (5 – 9.99 tons ha-1 y-1). models can be used to quantitatively determine the
According to them, the only part of the watershed amount of soil loss from watersheds and also to
landforms having very slight class of soil loss (0 - identify critical soil loss source areas (De Roo,
4.99 tons ha-1 y-1) are the flat plains, undulating 1998; Emrah et al., 2007). The successful
plains and the flat-flood prone areas; and followed application of such models, however, depends on
by slight soil loss (5 – 9.99 tons ha-1 y-1) for the the availability and quality of data for calibration
very steep escarpment of the watershed; and and validation (De Roo, 1998; Stefano et al., 1998;
moderate soil loss class (10 – 24.99 tons ha-1 y-1) Takken et al., 1999). Such problems are more
on rolling to hill landforms of the watershed, pronounced in developing regions where data
where as severe class of soil loss (25 – 44.99 tons availability is scarce, existing data are not easily
ha-1 y-1) was estimated using USLE on slopes 30- accessible and data collected and stored are mostly
50% (Table 4). This doesn’t mean that to give less in different formats. In addition, more complex
attention to those landforms with very slight to models do not necessarily perform better for
slight soil loss classes in the study watershed but watershed-scale management purposes, mainly
this is to indicate that parts of the watershed because input errors can increase with increasing
landforms that need high priority for SWC model complexity (Favis-Mortlock, 1998; Mitas
implementation using the available existing and Mitasova, 1998a; Jetten et al., 2003; Merritt, et
resource. This is because; it may be worth noting al., 2003).
that nature takes 200–400 years to build up 1 cm
of top soil (Pimental 1995) but thousands tons of Empirical models are frequently used in
soil are lost in a season from a watershed . He also preference to complex physically based models as
reported that each millimeters of cultivated soil they can be implemented in situations with limited
loss could cost 10 kg of nitrogen and 2 kg of data and parameter inputs, particularly as a first
phosphorus per ha. Hence, this study suggests for step in identifying sources and rate of soil loss
effective control of soil erosion at specific area (Merritt et al., 2003). However, such models

63
Marsland Press
Journal of American Science 2009: 5(1), 58-69
______________________________________________________________________________________
cannot be directly applied to environments other watershed. These activities are: watershed
than those for which they were developed, and treatment as area enclosure, afforestation, trench;
extrapolation of results from larger-scale plot-level reclamation of big gullies using check dams,
to small-scale watershed level application is biological; moisture harvesting techniques on
difficult. It is, therefore, necessary to identify farms and degraded grazing lands like soil, stone
models that are not very much simplified and and trench bunds; and soil faced stone bund on
under-represent the physical basis or not too hillsides.
complicated and very expensive to implement. The
best example is USLE, which is identified and fit Soil loss in different landforms of the
to apply in the case of the study area of Medego study watershed is influenced by erosion factors
watershed, northern Ethiopia. The USLE is an differently. For instance, the soil erodibility (K)
empirically based model developed in the United factor of the landforms in the watershed is a
States by using data on soil erosion rates. This function of soil texture, drainage condition and soil
equation has certain limitations but still is the best depth. These sub-factors can influence the soil
available method which is used most widely for color, which determined the value of K-factor in
estimating soil losses as average annual mass per USLE, adapted from Hurni (1985). The landforms
unit area as a function of the major factors in the watershed have different in texture, drainage
affecting sheet and rill erosion in data scarce areas condition, soil depth, soil color, land cover,
of developing countries. As all landscape positions erosion controlling management practices and
are not equally sensitive to erosion, one important slope factors (Table 3). Fine texture soils are
approach to tackling the problem of erosion could dominated on flat land areas where as coarser
be identifying where the sources of most of the soil textural class increases with increasing steepness.
loss are in a watershed (Dickinson and Collins, The same trend was observed for the soil depth
1998; Kim et al., 2007). Identification of potential with deeper soil on flat areas and shallow soil on
areas of erosion for appropriate management high slope gradient landforms. The drainage
interventions to tackle the major causative factors condition is extremely high on steeper landforms
at their specific locations is, therefore, imperative and poor on flat area of the watershed. Therefore,
from an economic, management and sustainability the principle of Hudson (1992) that describe as
point of view. This study was attempted in fine soil particles resist to detachment by raindrops
indicating the areas or landforms of high soil loss but they are susceptible to transport easily is soil
in Medego watershed, northern Ethiopia. drainage dependent. This is because if the
landform is poor in drainage, so the probability of
4.2. Soil Loss and the Influencing Factors in transporting by waters the fine particles long
Medego Watershed distance leaving the original area is too low.
Transportation and deposition processes are almost
It is a fact that environmental degradation balanced in such occasions. Drainage is affected
has been a problem in Tigray region, northern by the slope factor. That is why soil loss estimated
Ethiopia. The land surfaces in the region is mainly on flat landform is below the minimum tolerable
a reflection of the past erosion processes. The main soil loss (2 ton ha-1 yr-1) determined by Hurni
causes of soil erosion in the area among others (19985) for Ethiopia condition. This is the lowest
were out lined by different researchers ([Hurni, soil loss as compared to the other landforms in the
1985; Gebresilassie, 1996, Tilahun, 1996; Tamene, watershed (Table 4). Sand dominant soil textures
2005) and even witnessed by farmers as over- are common on higher slopes of the watershed.
cultivation, deforestation, over grazing, steep Even though they are coarser to transport as
topography, high rainfall intensity, unwise land compared to clay texture due to high soil drainage
use and management. This is evident by the huge condition of steep slopes, they are susceptible to
amount of soil loss, by water erosion and very low erosion in the watershed.
productivity of the farm lands. Therefore, to
rehabilitate the environment and enrich it to a Of course, the management practices in
meaningful level, a concerned effort on SWC the watershed also play its own great role in the
program has been carried out by the community magnitude of soil loss. Landforms with well land
coordinated by of bureau of agriculture under the cover indicated less soil loss. Because it dissipates
umbrella of the Tigray Regional Government, the energy from rain drops and also decreases the
northern Ethiopia. In the name of SWC program, volume and velocity of runoff effect. Soil loss
various types of physical and biological SWC estimated from landforms with very step slope (>
measures have been undertaken in the study 50%) in the study watershed is smaller than slopes

64
Estimating Soil Loss Using Universal Soil Loss Equation (USLE) for Soil Conservation planning at Medego Watershed, Northern
Ethiopia
Gebreyesus Brhane et al.
______________________________________________________________________________________
in the rage of 15-30% and 30-50%. The reason is as inter-bund management, bund stabilization,
cover factors and land managements factor are buffer zone establishment and re-bank re-
better in the very steeper slopes of the watershed. vegetation; and gully control by both vegetative
This includes less human and livestock and structural measures should be being
interferences, intensive terraces and relatively intensively implemented.
better vegetation cover of bushes and shrubs.
Therefore, the overall implication of this study is As a result of the implementation of
that after the implementation of SWC measures the SWC, the hydrological behavior of the watersheds
amount of soil loss in a given land unit is is improved such as base flow in streams and
decreased in many parts of the landforms by more springs increased, sediment load to reservoirs
than 50% in the watershed as compared to the high reduced, crop yield improvement due to soil
values indicated in the past studies in northern moisture enhancement, vegetation cover
Ethiopia (e.g., Hurni, 1985; 1990; 1993; Tekeste improvement and increased availability of forage
and Paul, 1989; Gebreselasie, 1996)). However, for livestock were observed. These are some of the
the present soil loss amount has also a significant indicators of the effectiveness of the implemented
influence on the overall productivity of the study soil and water conservation practices in the study
watershed unless the correct measures on the watershed. However, maintenance of the existing
targeted landforms are undertaken. This is because SWC and introducing additional appropriate land
as compared to the soil formation in the region management practices and rules should be given
which is not more than 2 ton ha-1 yr-1 (Hurni, 1985); attention by concerned bodies in order to decrease
the present soil loss estimated in Medego and totally stop the rate of soil loss and then to
watershed, northern Ethiopia is not neglected or it increase the total biomass production in the
is very big. Therefore, base on the landforms watershed, even though the biggest rate (35.43
identified in this study, soil conservation planning tons ha-1 yr-1) of soil loss is coming from landforms
should be undertaken to address the problem of having slopes 30-50% and the lowest soil loss is
erosion in areas having large soil loss as areas of from slope less than 2%. Therefore, to maximize
prioritization in the future. the available resources in targeting the effect of
water erosion on soil loss, those landforms and
land uses having large rate of erosion should be
5. CONCLUSION given first priority during the introduction of
The entire watershed area experienced intensive intensive and well designed SWC interventions at
rainfall which coupled with steep gradient slopes, Medego watershed, northern Ethiopia.
cause highly erosive runoff as in many other arid
and semi-arid areas of Ethiopia. It is this high Acknowledgements
runoff and soil detachment that is responsible for This research was conducted with the financial
the high rate of soil erosion at Medego watershed, support provided by Ethiopia Science and
northern Ethiopia that range from 1.59 – 35.43 Technology Agency Research Support Program
tons ha-1 y-1. There is a need to regulate this soil and Axum University, Ethiopia. The authors
loss by all possible means so as to decrease the thanks for the financial support of the
existing amount of soil loss and enhancing organizations.
watershed rehabilitation and productivity.
Suggested watershed rehabilitation as long and Corresponding author
short-term measures should be included the
following: As long-term measures re-vegetation of Gebreyesus Brhane
denuded hill slopes with trees and perennial Center for Development Research (ZEF),
grasses such as vetiver strips and belts; University of Bonn, Walter-Flex-Str. 3,
introduction of an agro-forestry program that is D-53113 Bonn, Germany
compatible with crop, livestock; and forestry phone: ++49 (0)162 4722826; Fax: ++49 (0)228-
development; where as short-term soil and water 73-5097;
conservation measures are given due attention to: e-mail: gebre042001@yahoo.com
cut-off drains which need to construct that
intercept runoff; constructing and maintenance
stone and soil bund and trenches on proper slopes
and soils and integrating with vegetation
intensively. This has to include interventions such

65
Marsland Press
Journal of American Science 2009: 5(1), 58-69
______________________________________________________________________________________
REFERENCES 14. Hurni H. Erosion - Productivity -
1. Bartsch KP, Van Miegroet H., Boettinger J, Conservation Systems in Ethiopia.
Dobrwolski, JP. Using empirical erosion Proceedings 4th International Conference on
models and GIS to determine erosion risk at Soil Conservation, Maracay, Venezuela 1985:
Camp Williams. Journal of Soil and Water 654-674.
Conservation 2002; 57(1):29–37. 15. Hurni H. Degradation and conservation of soil
2. Boardman J. An average soil erosion rate for resources in the Ethiopian highlands.
Europe: myth or reality? Journal of Soil and Mountain and Research Development 1990;8
Water Conservation 1998; (2)53:46-50. (2-3): 123-130.
3. BoARD, Bureau of Agriculture and Rural 16. Hurni H. Land degradation, famine, and land
Development. Annual report. Tigrigna (local resource scenarios in Ethiopia. In: Pimentel D.
language) version. Axum, Ethiopia. 2007: 4- (Ed.) World soil erosion and conservation.
18. Cambridge Univ. Press. 1993:89-97.
4. Chekun T. Vetiver in the Rehabilitation of the 17. Jetten V, Govers G, Hessel R. Erosion models:
Degraded ZegZeg Watershed in Ethiopia, quality of spatial predictions. Hydrological
Addis Ababa, Ethiopia. Report, 2002:131-136. Processes 2003;17(5):887 – 900.
5. De Roo APJ. Modelling runoff and sediment 18. Judson S. Physical Geology, Prentice Hall, NJ,
transport in catchments using GIS. USA, 3rd ed., 1965:143–144
Hydrological Processes 1998;12:905-922. 19. Kim M, Flanagan DC, Frankenberger JR,
6. Emrah HE, Günay E, ølhami B. Use of Jung K.. Regional Scale Application of USLE
USLE/GIS Methodology for Predicting Soil and WEPP for Soil Erosion Assessment in
Loss in a Semiarid Agricultural Watershed. Korea [abstract]. 2007 American Society of
Environmental Monitoring and Assessment Agricultural and Biological Engineers Annual
2007;131(1-3):153–161. International Meeting. June 17-20, 2007,
http://www.springerlink.com/content/rx032v5 Minneapolis, MN. Paper No. 072048.
3753w4367/fulltext.pdf.Accessed 18 Dec. 20. Lal R. Erosion Impact on Soil quality in the
2008. Tropics. In Lal (Ed.): Soil Quality and Soil
7. Favis-Mortlock DT. Validation of field-scale Erosion. CRC press, Boca Raton, Florida.
soil erosion models using common datasets. In: 1999: 285-317.
Boardman J and Favis-Mortlock DT (Eds.) 21. Machado MJ, Perez-Gonzalez A, and Benito
Modelling Soil Erosion by Water, Springer- G. Geomorphology and soils. In: Feoli E (Ed.)
Verlag NATO-ASI Series I-55, Berlin. 1998: Rehabilitation of degrading and degraded
89-128. areas of Tigray, northern Ethiopia.
8. Ferro V, Porto P, Tusa G. Testing a Department of Biology, University of Trieste,
distributed approach for modeling sediment Trieste, 1996:65-106.
delivery. Hydrological Sciences Journal 1998; 22. Merritt WS, Letcher RA, and Jakemna AJ. A
43(3): 425-442. review of erosion and sediment transport
9. FAO/UNEP. Methodology for Assessing Soil models. Environmental Modelling and
Degradation, FAO/UNEP, New York, Software 2003;18(8-9): 761-799.
1978:49–55. 23. Mirco G, Robert JA.J, Ezio R, Luca M. Soil
10. FAO. Ethiopian Highland Reclamation Study. Erosion Risk in Italy: a revised USLE
Ethiopia. Final report. FAO. Rome. 1986:37- approach. European Soil Bureau Research
46. Report No.11, EUR 20677 EN. Office for
11. Gebreselasie ED. Soil erosion hazard Official Publications of the European
assessment for land evaluation. Soil Communities, Luxembourg. 2003:28.
Conservation Research Program, 24. Mitas L, Mitasova H. 1998 Distributed soil
University of Bern, Switzerland and the erosion simulation for effective erosion
Ministry of Agriculture, Ethiopia. MSc Thesis, prevention. Water Resources Research.
1996:68-82. 1998;34(3): 505-516.
12. Hudson N. Land Husbandry. B T Batsford 25. Phillips JD. Hillslope and channel sediment
Limited, London. 1992:172-190. delivery and impacts of soil erosion on water
13. Hurni H. Soil erosion and soil formation in resources. In Hadley RF and Ongley EO
agricultural ecosystems in Ethiopia and (Eds.) Sediment and the Environment IAHS
Northern Thailand. Mountain research and Publ. 1989;23(184):183-190.
development 1983; 3(2):131-142.

66
Estimating Soil Loss Using Universal Soil Loss Equation (USLE) for Soil Conservation planning at Medego Watershed, Northern
Ethiopia
Gebreyesus Brhane et al.
______________________________________________________________________________________
26. Pimental DC. Environmental and economic 32. Takken I, Beuselinck L, Nachtergaele J,
cost of soil erosion and conservation benefits. Govers G, Poesen J, Degraer G. Spatial
Science 1995;267(5201):1117–1123. evaluation of a physically-based distributed
27. Renard KG, Foster GR. Soil conservation: erosion model (LISEM). Catena 1999;37(3-4):
principles of erosion by water. In: Degne HE 431-447.
and Willis WO (Eds.) Dryland Agriculture, 33. Tamene L. Reservoir siltation in the drylands
Agronomy Monogr. 23, Am. Soc., Crop Sci. of northern Ethiopia: causes, source areas and
Soc. Am., and Soil Sci. Am Madison, management options. PhD Thesis, Ecology
Wisconsin, 1983:156-176. and Development Series 30, Center for
28. Renard KG, Foster GR, Weesies GA, McCool Development Research, University of Bonn.
DK, Yoder DC. Predicting soil erosion by 2005:84-138.
water: a guide to conservation planning with 34. Tekeste G, Paul DS. Soil and water
the Revised Universal Soil Loss Equation conservation in Tigray, Ethiopia. Report of a
(RUSLE). USDA Agricultural Handbook 703, consultancy visit to Tigray.Univ. Wageningen.
1997:126-131. 1989:78.
29. Singh R, Phadke VS, 2006. Assessing soil loss 35. Tilahun Y. Impacts of conservation bunds on
by water erosion in Jamni River Basin, crop yields in Degua Tembien, northern
Bundelkhand region, India, adopting universal Ethiopia. MSc Thesis, Agricultural University
soil loss equation using GIS. Current Science, of Norway (NLH). 1996: 53-82.
2006; 90(10): 25 -27. 36. Wischmeier WH, Smith DD. Predicting
30. Stefano CD, Ferro V, Porto P. Linking rainfall erosion losses – a guide for
Sediment Yield and Caesium-137 Spatial conservation planning. U.S. Department of
Distribution at Basin Scale. Journal of Agriculture, Agriculture Handbook 537.
Agricultural Engineering Research 1998;74 1978:20-152.
(1): 41-62. 37. Zeleke G, Hurni H. 2001. Implications of land
31. Sutcliffe JP. Economic assessment of land use and land cover dynamics for mountain
degradation in the Ethiopian highlands: A case resourcedegradation in the Northwestern
study. National Conservation Strategy Ethiopian highlands. Mountain Research and
Secretariat, Ministry of Planning and Development 2001;21(2): 184-191.
Economic Development, Addis Ababa, 1993.

Fig 1: Map of Ethiopia with Tigray region and Medego watershed outlined.

67
Marsland Press
Journal of American Science 2009: 5(1), 58-69
______________________________________________________________________________________

68
Estimating Soil Loss Using Universal Soil Loss Equation (USLE) for Soil Conservation planning at Medego Watershed, Northern
Ethiopia
Gebreyesus Brhane et al.
______________________________________________________________________________________

69
Marsland Press
Journal of American Science 2009: 5(1), 70-76
______________________________________________________________________________________
Research Article
Conservation through in vitro method: A case of plant regeneration
through somatic embryogenesis in Quercus semecarpifolia Sm.

1
Sushma Tamta , 2 Lok Man S. Palni, 2P. Vyas and 2M.S.Bisht
1
Botany Department, D. S. B. Campus, Kumaun University, Nainital, UA, 263 002, India.
2
G.B.Pant Institute of Himalayan Environment and Development, Kosi-Katarmal, Almora, UA, 263
643, India
____________________________________________________________________
An efficient and reproducible protocol for in vitro propagation via somatic embryogenesis (direct as
well as indirect) induced on cotyledon halves (with embryo) taken from seeds of Quercus
semecarpifolia (Sm.) has been developed. Direct as well as indirect somatic embryogenesis was
induced from the cotyledons on Woody plant (WP) medium supplemented with 6-Benzyladenine (BA)
+ Indole-3-butyric acid (IBA), and, BA + 2,4-Dichlorophenoxyacetic acid (2,4-D), respectively.
Somatic embryos thus obtained were multiplied profusely on Schenk and Hildebrandt (SH) +
Murashige and Skoog (MS) basal as well as BA supplemented media. Germination and conversion of
somatic embryos into plantlets was achieved on SH+MS medium supplemented with BA (0.44- 8.88
PM). Rooting of in vitro produced shoots was achieved on WP (1/2 macro + full concentration of rest
of the constituents) medium supplemented with IBA (14.76 PM). The plants were hardened ex-vitro
and transferred to earthen pots containing garden soil. [Journal of American Science 2009: 5(1), 70-76]
(ISSN: 1545-1003)

Key words: Quercus semecarpifolia, brown oak, somatic embryogenesis, micropropagation.

regeneration have recently become available for

1. INTRODUCTION many plant species, including Arabidopsis
thaliana, a model plant in genetics and
The genus Quercus has a wide distribution embryogenesis (Gaj, 2004).
range; mostly trees, either deciduous or evergreen
and is of enormous ecological and economical 2. MATERIALS AND METHODS
value. One of the species of Quercus, i.e., Quercus
semecarpifolia Sm. (family-Fagaceae); common Plant material and surface sterilization
name-brown or kharsu oak; is the main forest
forming evergreen tree species around 2400 m Seeds of Quercus semecarpifolia Sm. were
amsl in parts of Indian Himalaya (Singh and Singh, collected from well grown adult tree in the natural
1987). In view of the general importance of this forests at Kilbury, Nainital (2100-2400 m amsl;
species and problems associated with its 29q 24c 30s N- 29q 27c N lat. and 79q 25c E- 79q
regeneration (Tamta et al. 2008), in the present 29c 40s E long.), Uttarakhand, India. Following
study attempt has been made for the first time to surface disinfection (Tamta et al. 2008), the seed
develop an efficient in vitro micropropagation coat was removed and seeds were divided into two
method through somatic embryogenesis. halves; one half containing only one cotyledon
while the other half contained the other cotyledon
Micropropagation through somatic along with the embryo. These seed halves were used
embryogenesis offers considerable advantages as explants for inoculation.
over other methods of clonal propagation; this
route has a high proliferation potential. It has been Media and culture establishment
considered as a very promising method of oak
micropropagation (Chalupa, 1995, Wilhelm 2000, Three basal media, namely MS (Murashige
Purohit et al.2002), and was found to be highly and Skoog, 1962), WP (Lloyd and McCown, 1980)
reproducible in this study on Q. semecarpifolia. and SH+MS, i.e., a combination of macronutrients
Efficient protocols on SE induction and plant

70
Conservation through in vitro method: A case of plant regeneration through somatic embryogenesis in Quercus semecarpifolia Sm.
Sushma Tamta et al.
______________________________________________________________________________________
of SH (Schenk and Hilderbrandt, 1972) and the After 5 weeks, the shoots with well developed
remaining constituents of MS, were used. The basal roots were taken out from the culture flasks, the
media were supplemented with various roots gently washed with water to remove traces of
concentrations of auxins, cytokinins and phytagel and the plantlets were then transferred to
gibberellins. The sucrose concentration was 3.0% small plastic cups (8.0 cm ht; 7.0 cm dia) containing
(w/v) and the media were solidified with 0.8% agar garden soil and the cups were covered with a
(w/v). The experiments were done using glass transparent polythene sheet. Plants were kept inside
petridishes (10 cm dia, 25 ml medium per petirdish) a polyhouse for acclimatization for 1 month. After
or conical flasks (250 ml volume, 100 ml medium that the plants were transferred to the earthen pots
per flask). Incubation of cultures was carried out at (18 cm high; 20 cm dia) containing the same soil.
25 ±1 °C in a 16 h light and 8 h dark cycle, with 42.0
µmol m -2s-1 and 60.0 µmol m -2s-1 irradiance inside Statistical analyses
and outside the culture flasks, respectively by cool
fluorescent tubes (Philips TI 40 W/54). Experiments were conducted using a
randomized block design to determine the effect of
Production of somatic embryos treatments and were repeated as described in
individual experiments. For all the experiments
Seed halves turned green when inoculated explants were used in triplicates.
on WP basal medium. After seven days,
cotyledons with or without the zygotic embryo, 3. RESULTS AND DISCUSSION
were transferred on to WP or MS medium
supplemented with either BA (0.44 PM) alone or Direct somatic embryogenesis
in combination with 2,4-D (4.53 PM) or IBA (4.92
PM) or GA3 (2.89 PM). Direct as well as indirect Globular structures were found to develop
somatic embryogenesis with the intervening callus directly on the periphery of cotyledons with
phase was induced within 13 weeks and 18 (10 attached zygotic embryo, after 13 weeks on WP
weeks for callus establishment and proliferation + medium supplemented with BA and IBA (Table 1).
8 weeks for induction of somatic embryos) weeks These structures were loosely attached to the
of culture, respectively. In both the cases, the surface of cotyledons (Fig. 1A). On subculture
presence of zygotic embryo seems to have some these globular structures were converted into
role in the production of somatic embryos. The bipolar somatic embryos (Fig. 1B). This has been
callus raised from cotyledons without the zygotic reported in some other species of Quercus
embryos did not survive on further subculture and (Chalupa, 1995; Gingas and Lineberger, 1989).
degenerated. For germination of somatic embryos, Bipolar somatic embryos were also observed in Q.
formed both from the direct as well as indirect robur (Cuenca et al., 1999) and in Q. suber
pathways, SH+MS medium supplemented with (Puigderrajols et al., 1996), which were reported to
BA (0.44-8.88 PM) was used. The somatic be translucent or opaque-white in appearance.
embryos germinated to form well developed These somatic embryos were multiplied by
shoots, leaves and tap root system. secondary embryogenesis (Fig. 1C), and the
frequency of secondary embryo formation was
Adventitious rooting of microshoots found to increase when subcultured on SH+MS
medium, without any growth regulators. In Q.
The survival rate of plantlets thus obtained; suber also secondary embryogenic lines were
after transfer to ex vitro conditions was very poor mainted on medium lacking PGRs (Fernandez-
(data not shown). Therefore, the main tap root was Guijarro et al., 1995). Proliferation of secondary
excised and the shoots were transferred to the embryos was most prolific from the root pole of
rooting medium, i.e., WP (1/2 macro + full the somatic embryos. Secondary embryos were
concentration of rest of the constituents) or SH + MS produced mostly from the root pole end of the
(macro of SH + rest of the constituents of MS) primary embryos as also observed by El Maataouti
media supplemented with different auxins (Table 4), et al. (1990) and Gingas (1991). Cotyledons
containing sucrose (3.0%; w/v) and phytagel (0.25%; without the embryonic axes failed to give rise to
w/v). Well developed adventitious roots were found direct embryos.
to form within 4 weeks.
Indirect somatic embryogenesis
Transfer of plantlets to soil

71
Marsland Press
Journal of American Science 2009: 5(1), 70-76
______________________________________________________________________________________
Callus was induced from the surface of conversion of somatic embryos into full plants in
cotyledons inoculated on both MS or WP media oaks is usually quite low (Chalupa, 1995); this is a
supplemented with BA and 2,4-D or IBA (Table 1). matter of future investigations. Fig. 1H shows the
The creamy yellow callus developed on MS medium germination of somatic embryo.
was slow to proliferate and degenerated on further
subcultures. On the other hand friable callus was Adventitious rooting of microshoots excised from
formed on WP medium after 10 weeks on germinating somatic embryos
cotyledonary halves with embryo (Fig. 1D);
subsequently this callus was subcultured on MS Out of various media tried (MS, WP,
basal medium (half or full strength) supplemented SH+MS) supplemented with various auxins (IAA,
with CH (0.02%, w/v) and activated charcoal (1.0%, NAA, IBA) in different concentrations (4.92 PM -
w/v) (Table 2). The friable callus developed on WP 28.55 PM), WP medium supplemented with IBA
medium supplemented with BA (0.44 PM) and (14.76 PM) was found to be most effective
2,4-D (4.53 PM) (Table 1) was found to turn (100.0%) in inducing rooting without any callus
embryogenic after 8 weeks (two months) of formation at the basal end (Table 4). The root
subculture (Table 2; Fig. 1E) when transferred to the initials were observed within 10 days and well
above medium, i.e., MS basal (half or full strength) developed roots were formed in four weeks (Fig. 1I).
medium supplemented with CH (0.02%; w/v) + AC The average number of roots was 12.46 with
(0.1%; w/v). Somatic embryos could be multiplied maximum length of 6.97 cm (Fig. 1J). WP medium
through secondary embryogenesis on SH + MS supplemented with NAA or IAA also induced
medium supplemented with BA (0.44-8.88 PM) rooting (16.6% and 50.0%, respectively). However,
(Table 3). BA, a potent cytokinin, alone or in the average number of roots was 3.0 and 3.02 and
combination with auxins, particularly IBA or 2,4-D, the length of the longest roots were 0.2 and 2.2 cm,
has been known to induce somatic embryogenesis respectively. When IBA was added to SH+MS
from the zygotic embryos (Chalupa, 1995; Gingas medium, this combination also induced rooting
and Lineberger, 1989; Sasamoto and Hosoi, 1992; (100.0%) but the formation of callus was invariably
Kim et al. 2006). Somatic embryos of all stages seen at the base of the explant, and the average
(globular, heart and torpedo shaped) could be number of roots (4.3) and length of the longest root
observed on the same medium. (0.21 cm) were also considerably less. The addition
of NAA to SH+MS medium totally failed in
The rate of multiplication of somatic inducing rooting, whereas IAA induced rooting in
embryos through secondary embryogenesis varied 40.0% shoots with the average number roots being
from 1.66 to 3.14 secondary embryos per somatic 4.0. However, the roots did not elongate and the
embryo, over a period of 5-6 weeks, depending length of the longest root never exceeded beyond
upon the PGR supplements (Table 3). It is often 0.2 cm. Secondary roots were found to develop only
reported in case of Quercus that calli turn on WP medium supplemented with IBA with
embryogenic when transferred to the basal medium profuse adventitious rooting. Addition of IBA to the
(Gingas and Lineberger, 1989; Guijarro et al., 1995; rooting medium gave better results in comparison to
Kim et al., 1994). another auxin, NAA, in Q. suber (Manzanera and
Pardos, 1990) also.
Germination of somatic embryos
Hardening: Well rooted plants were taken out
Somatic embryos (produced from the direct of the culture vessels and the adhering phytagel was
as well as indirect pathways) were transferred to BA carefully removed; the delicate roots were then
(0.44-8.88 PM) supplemented SH+MS medium for gently and thoroughly washed before transferring to
germination. Some of the somatic embryos plastic cups containing garden soil (Fig. 1K). The
germinated and produced root and shoot in a well survival of these plants was only 20.0 per cent.
coordinated manner (Fig. 1F). In a number of After one month, these plants were transferred to
somatic embryos only the root primordia elongated earthen pots containing same soil and maintained
(Fig. 1G); its frequency varied from 4.0-27.0 per inside the polyhouse until new leaves were found to
cent depending upon the concentration of BA in the emerge (Fig. 1L). In conclusion, the present study
medium. The overall conversion frequency of describes, for the first time, the effective
somatic embryos was only around 10 per cent. BA multiplication protocol for in vitro propagation of Q.
at 2.22 PM was found to be optimum for semecarpifolia.
germination and conversion of somatic embryos
into plantlets (Table 3). The frequency of

72
Conservation through in vitro method: A case of plant regeneration through somatic embryogenesis in Quercus semecarpifolia Sm.
Sushma Tamta et al.
______________________________________________________________________________________

Table 1
Effect of treatments on seed halves of Q. semecarpifolia in different media

S. No. Treatments MS medium WP medium

1 Control - -
2 BA (0.44 PM) - -
3 BA+2,4-D (0.44 PM+4.53 PM) Callus Callus*
4 BA+IBA (0.44 PM+4.92 PM) Callus Direct SE
5 BA+ GA3 ((0.44 PM+2.89 PM) - -

*embryogenic callus, - nil, SE: somatic embryogenesis, data recorded after 10 weeks of culture for callus
formation and after 13 weeks for direct somatic embryo formation

Table 2
Callus proliferation and somatic embryogenesis in Q. semecarpifolia

Medium constituents Callus Embryogenesis No. of embryos/

Proliferation petri dish

MS ++ *** 125
MS+CH (0.02%) +++ ** 96
MS+CH (0.02%) +AC (0.1%) + - NA
1/2 MS + CH (0.02%) + - NA
1/2MS+CH (0.02%) + AC (0.1%) ++ *** 110

The callus was initiated on WP medium supplemented with BA and 2,4-D;

MS: Murashige and Skoog medium; CH: Casein hydrolysate, AC: activated charcoal, all concentrations are
w/v basis; + poor, ++ medium, +++ prolific; * poor, ** moderate, *** abundant, - nil, NA: not applicable;
data recorded after 8 weeks (2 months) of culture; 6 petridishes were used per treatment with 4 callus pieces
per petridish; the experiment was repeated twice with similar results

73
Marsland Press
Journal of American Science 2009: 5(1), 70-76
______________________________________________________________________________________
Table 3
Response of somatic embryos of Q. semecarpifolia on SH+MS medium supplemented with various
concentrations of BA

BA No. of somatic Germination of Secondary Frequency of root

(PM) embryos somatic embryos (%) embryogenesis* formation (%)
transferred
0.44 30 0 1.66 26.60
0.88 97 4.50 2.28 18.40
1.78 44 5.20 3.14 18.18
2.22 49 6.90 3.00 14.00
4.44 68 2.90 1.85 4.40
8.87 90 0.89 1.76 4.10

*
No. of total somatic embryos after six weeks/no. of somatic embryos initially inoculated per flask; each
treatment consisted of 12 flasks, data was recorded 6 weeks after transfer of somatic embryos to the medium.
The experiment was repeated twice with similar results.

Table 4
Effect of auxins and media on in vitro rooting of SE derived
microshoots of Q. semecarpifolia

Medium PGRs Shoot ht % % No. of roots/ Length of Sec.

(conc.in PM) (cm) rSD callusing rooting shoot rSD longest root roots
(cm)rSD

WP IBA (14.76) 2.20 r1.04 0.00 100.00 12.46 r4.87 6.97 r1.47 +
NAA(16.11) 1.33 r0.68 100.00 16.60 3.00 r1.22 0.20 r0.03 -
IAA (17.13) 2.17 r0.69 0.00 50.00 3.02 r4.24 2.20r 0.57 -
SH+MS IBA (14.76) 2.56 r0.42 100.00 100.00 4.30 r2.07 0.21 r0.13 -
NAA (16.11) 1.93 r0.89 48.00 0.00 NA NA NA
IAA (17.13) 2.00 r1.31 0.00 40.00 4.00 r2.3 0.20 r0.11 -

WP: 1/2 macro + full concentrations of rest of the constituents; SH+MS : macro (SH) + rest of the
constituents of MS; SE: somatic embryo, +: occurred; -: did not occur; NA: not applicable; SD: standard
deviation, data recorded 5 weeks after transfer to rooting medium, treatments were carried out in triplicate and
each flask contain 9 microshoots

74
Conservation through in vitro method: A case of plant regeneration through somatic embryogenesis in Quercus semecarpifolia Sm.
Sushma Tamta et al.
______________________________________________________________________________________

A B C

D E F

G H I

J K L

Fig. 1. In vitro propagation of Q. semecarpifolia

(A) Globular structures loosely attached to the surface of the cotyledon.
(B) Bipolar somatic embryos
(C) Secondary embryogenesis
(D) Friable embryogenic callus on WP medium
(E) Indirect somatic embryogenesis
(F) Germination of somatic embryo
(G) Elongation of root primordial from the somatic embryo
(H) Different stages of somatic embryo germination
(I) Well rooted plantlets after 4 weeks of culture on WP medium
supplemented with IBA
(J) Rooting from basal view
(K) Well rooted plant 1 month after transfer to plastic cup containing
garden soil
(L) Two –months-old in vitro propagated plant in earthen pot

75
Marsland Press
Journal of American Science 2009: 5(1), 70-76
______________________________________________________________________________________

ACKNOWLEDGEMENT 9. Kim YW, Lee BC, Lee SK, Jang SS (1994)

The financial support received from the Department of Somatic embryogenesis and plant regeneration
Biotechnology, Govt. of India, is gratefully in Quercus acutissima. Plant Cell Rep. 13:
acknowledged. 315-318
10. Kim Y, Moon H, Son S (2006) Repetitive
somatic embryogenesis and plant regeneration
Corresponding Author: in Zizyphus jujube Mill. InVitro Cell Dev Bio
Dr (Mrs) Sushma Tamta, Plant. 42(3): 247-251
Lecturer, Botany Department, 11. Lloyd G. and McCown B. (1980)
D. S. B. Campus, Kumaun University, Nainital, Commercially feasible micropropagation of
Uttarakhand, 263 002, India. mountain laurel, Kalmia latifolia, by shoot tip
Email: sushma_tamta@yahoo.com culture. Comb. Proc. Intl. Plant Prop. Soc. 30:
Telephone:9412924956 421-427.
12. Manzanera J.A. and Pardos J.A. (1990)
Micropropagation of juvenile and adult
REFERANCES Quercus suber L. Plant Cell Tiss. Org. Cult.
1. Chalupa V (1995) Somatic embryogenesis in 21: 1 – 8.
oak (Quercus spp.). In: Jain S, Gupta P, 13. Murashige T. and Skoog F. (1962) A revised
Newton R (eds) Somatic embryogenesis in medium for rapid growth and bioassays with
Woody Plants. Angiosperms. Kluwer tobacco tissue culture. Physiol. Plant. 15:
Academic Publishers, Dordrecht, The 473 – 497.
Netherlands, vol. 2, pp 67 – 87 14. Puigderrajols P, Fernandez-Guijarro B,
2. Cuenca B, San-Jose MC, Martinez MT (1999) Toribio M, Molinas M (1996) Origin and
Somatic embryogenesis from stem and leaf early development of secondary embryos in Q.
explants of Quercus robur L. Plant Cell Rep. suber L. Int. J. Plant Sci. 157: 674-684
18: 538-543 15. Purohit V.K., Palni L.M.S., Nandi S.K. Vyas
3. El Maataoui M, Espagnac H, P. and Tamta S. (2002) Somatic
Michaux-Ferriere N (1990) Histology of embryogenesis in Quercus floribunda, an
callogenesis and somatic embryogenesis important Central Himalayan oak. In: Role of
induced in stem fragments of cork oak plant tissue culture in biodiversity
(Quercus suber) cultured in vitro. Ann. Bot. conservation and economic development. Eds.
66: 183-190 Nandi S.K., Palni L.M.S. and Kumar A.
4. Fernandez-Guijarro B, Celestino C, Toribio M Gyanodaya Prakashan, Nainital, pp. 41-52.
(1995) Influence of external factors on 16. Sasamoto H, Hosoi Y (1992) Callus
secondary embryogenesis and germination in proliferation from the protoplasts of
somatic embryos from leaves of Quercus embryogenic cells of Quercus serrata. Plant
suber. Plant Cell Tiss. Org. Cult. 41: 99-106 Cell Tiss. Org. Cult. 29: 241-245
5. Gaj DM (2004) Factors influencing somatic 17. Schenk R.U. and Hildebrandt A.C. (1972)
embryogenesis induction and plant Medium and techniques for induction and
regeneration with particular reference to growth of monocotyledonous and
Arabidopsis thaliana (L.) Heynh. Plant dicotyledonous plant cell cultures. Can. J. Bot.
Growth Regulation. 43(1): 27-47 50: 199-204
6. Gingas VM (1991) Asexual embryogenesis and 18. Singh JS, Singh SP (1987) Forest vegetation of
plant regeneration from male catkins of the Himalaya. Bot. Rev. 53 (1): 80-192
Quercus. Hort. Sci. 26: 1217-1218 19. Tamta S., Palni L.M.S., Purohit V.K, and
7. Gingas VM, Lineberger RD (1989) Asexual Nandi S.K. (2008) In vitro propagation of
embryogenesis and plant regeneration in brown oak (Quercus semecarpifolia Sm.)
Quercus. Plant Cell Tiss. Org. Cult. 17: 191- from seedling explants (accepted: 21 March
203 2008 for publication in In Vitro
8. Guijarro FB, Clestino C, Toribio M (1995) Cell.Dev.Biol.-Plant; 44(2):136-141.
Influence of external factors on secondary 20. Wilhelm E (2000) Somatic embryogenesis in
embryogenesis and germination in somatic oak (Quercus spp.). InVitro Cell Dev Bio
embryos from leaves of Quercus suber. Plant Plant. 36: 349-357
Cell Tiss.Org. Cult. 41: 99-106

76
Marsland Press
Journal of American Science 2009: 5(1), 77-82
___________________________________________________________________________________
Research Article
Synthesis, characterization and Electroluminescence of
BPh2(2-(benzimidazol-2-yl) pyridinato) compound

Anchi Yeh
Department of Chemical Engineering,
Chengshiu University, Kaohsiung, Taiwan, R.O.C.
E-mail: acyeh@csu.edu.tw
Phone number: 886-7-7247891
Fax number: 886-7-7247891
________________________________________________________________
ABSTRACT: A novel luminescent boron compound, BPh2(2-(benzimidazol-2-yl) pyridinato)
(B-BIP), have been synthesized by reactions of triphenylboron with appropriate ligands,
2-(2-pyridyl)benzimidazole (BIP). For the three-layer OLED with the structure ITO/NPB/
B-BIP /Alq3/Mg-Ag, an emission band covering the whole visible region from 400 to 650 nm
with the maximum brightness of 50 cd/m2 was observed, indicating a perfect white light OLED
(CIE = 0.32, 0.37). [Journal of American Science 2009: 5(1), 77-82](ISSN: 1545-1003)

Keywords: white light; electroluminescence; imidazole; boron;

1. INTRODUCTION been known as good chelating ligands [13] and

the attachment of the pyridyl group at 2-position
The chemistry of organoboron compounds have
of imidazole would allow the new ligand to form
attracted much more attention recently because
stable compounds with the other atoms. In the
they are of interest for practical applications
present work, the syntheses, structures, and
[1-3]. Since an organic light emitting diode
electroluminescent properties of two new boron
(OLED) was reported by Tang and Vanslyke [4],
compounds BPh2(2-(benzimidazol-2-yl)
LEDs based on organic materials have generated
pyridinato)(B-BIP) is reported.
considerable interest and enabled the
development of low-cost, full-color, flat-panel
displays [5-8]. The best-known EL metal
2. EXPERIMENTAL METHOD
complex used in OLED is Alq3 which is not only
The synthesis of the title compound was
a good emitter but also a highly efficient
accomplished by following processes, as shown
electron-transporting material, where q is the
in Scheme 1. The triphenylboron (1.45g, 6.0
8-hydroxyquinolinato ligand [9-12]. Via the
mmole) was slowly added to 100 ml of THF
modification of the ligands of metal complexes,
solution containing 2-(2-pyridyl)benzimidazole
the emission spectra of devices and other
(2.63g, 13.5 mmole) at 0oC under N2. After the
properties, such as thermo stability and carrier
resulting mixture was stirred at room
mobility, can be tuned. The imidazoles have

77
Synthesis, characterization and Electroluminescence of BPh2(2-(benzimidazol-2-yl) pyridinato) compound
Anchi Yeh
___________________________________________________________________________________
temperature for 6 hours, 5 ml isopropyl alcohol present wok, the
was added to quench the reaction. The solvents
N,N’-bis-(1-naphthyl)-N,N’-diphenyl-1,1’-biphe
were removed under vacuum condition at 5×10-3
nyl-4,4’-diamine (NPB) was used as the
Torr, and the residual solid was sublimed to
hole-transport material (HTM), and tris
purify the final product. Light green of B-BIP
(8-quinnolinolato) aluminum (Alq3) was
was obtained in 90% yield. The organic light
employed as the electron-transporting material
emitting device, Fig. 1, using B-BIP as the
(ETM). The EL spectrum and the Commission
emitting and electron-transporting layer were
Internationale de l’Eclairage (CIE) co-ordinates
fabricated on the transparent conductive
were measured by Pro-650 Spectroscanner (step
indium-tin oxide (ITO) glass substrate. The
size is 1.0 nm and bandpass is 4nm), the
organic layers and the cathode were sequentially
current-voltage (I-V) characteristic was
deposited by conventional vacuum vapor
measured by Keithley 2400 Source meter.
deposition in the same chamber without
Thermogravimetric analysis (TGA) was
breaking the vacuum under 3×10-6 Torr. The
performed on a Perkin-Elmer thermogravimeter
cathode composed of magnesium silver alloy
(Pyris 1) under a dry nitrogen gas flow at the
(Mg:Ag = 10:1) were deposited onto the top
heating rate of 20oC/min. Glass transition
layer of organic materials by co-evaporation of
temperature (Tg) and melting point (Tm) of
Mg and Ag from different source. Before the
materials were determined by differential
deposition, all of the organic materials were
scanning calorimetry of the Perkin-Elmer
purified by the train sublimation method. In the
differential scanning calorimeter (DSC-7).

N N

N
+ BPh 3 N
B
N
H

Scheme 1. Synthesis process for the title compound

Mg/Ag (2000Å)
Alq3 (500Å)
B-BIP (500Å)
NPB (500Å)
ITO
Substrate

Fig 1: Device structure of organic light emitting device (OLED) fabricated in this work

78
Marsland Press
Journal of American Science 2009: 5(1), 77-82
___________________________________________________________________________________

3. RESULTS AND DISCUSSION [14, 15]. The thin films of B-BIP used for the
analyses of UV-vis and photoluminescence
A new boron compound spectra were obtained by depositing B-BIP onto
BPh2(2-(benzimidazol-2-yl) pyridinato) (B-BIP) quartz substrates under vacuum condition. At
was prepared by reacting triphenylboron with room temperature and low concentration (1×10-5
appropriate imidazole in dry THF (Scheme 1). M), the absorption spectral features of B-BIP in
Both of the compounds are air-stable in the solid N,N’-dimethylformamide (DMF) consist of two
state and in solution. The Thermogravimetric discrete bands (Fig. 2 ). The strong absorptions
analyses (TGA) scans under nitrogen for B-BIP centered at 280 nm for B-BIP, respectively, can
powder showed weight loss of 10% at 301 oC, be assigned to the ʌ-ʌ* transition. The other
respectively, which reveal that B-BIP is quite intense band centered at 348 nm shows a
stable in the atmosphere of nitrogen. The DSC vibrational separation of 1000 cm-1 with the Ȟ0, 0
results indicate that the compound B-BIP transition at 2.94 × 103 cm-1. This lower energy
possess a very high melting temperatures, 289oC, band possesses a reasonably high
respectively, which may serve as an advantage absorptivty (İ Д 3×104 dm3mol-1cm-1)
for OLED device fabrication because the and a red shift with increasing polarity
materials having high transition temperature of solvent, which is typical for a ʌˀʌ*
could provide the device with greater longevity transition [16-18].

-5
B-BIP (1X10 M)
Absorbance (a.u.)

0.8

0.4

0.0

300 400 500

Wavelength (nm)

Fig 2: UV-vis spectra of B-BIP in N, N’-dimethylformamide

Fig. 3 show the photoluminescence (PL) spectra with the configuration of

of the solutions and neat film of B-BIP excited
ITO/NPB/B-BIP/Alq3/MgAg was fabricated by
with 355 nm laser line. All concentration in
using NPB as the hole-transporting layer and
DMF, only one emission band was observed
B-BIP as the emitter and Alq3 is
with a maximum at 455 nm. Compound B-BIP
electron-transporting layer. The EL spectrum of
possess the appreciable PL quantum yield, with
organic light emitting device at the bias voltage
ͅf = 0.66 respectively, 10-6 M in DMF relative
of 10 V, Fig. 4, shows the broader emission
to
bands ranging from 400 to 650 nm were
3-(2-benzothiazolyl)-7- diethyl-aminocoumarin observed, indicating that the three-layer LED
(C540). To investigate the electroluminescent device emitted white light covering the whole
properties of B-BIP typical three-layer device visible light region. The band around 455 nm in

79
Synthesis, characterization and Electroluminescence of BPh2(2-(benzimidazol-2-yl) pyridinato) compound
Anchi Yeh
___________________________________________________________________________________
EL spectrum can be attributed to emission of emission is very important because this kind of
B-BIP, because its emission position is almost material is very seldom prepared so far, and it is
identical with that in PL spectrum of B-BIP. The very important for the fabrication of display
emission band at 535 is Alq3 emission position. panels. At the same time important role here
The emission is almost fixed in the white region may play electron-vibration interactions
in the CIE coordinate of x = 0.32 y = 0.37. For determining the spectral broadening of the
the small molecular organic materials, to emission lines. So the future strategy of the
develop the double layer of device with white materials design may be in this way also.

-3
bimp-B(1X10 M)
1.0 -4
bimp-B(1X10 M)
-5
bimp-B(5X10 M)
thin film
Normalize Intensity

0.8

0.6

0.4

0.2

0.0
400 500 600
Wavelength(nm)

Fig 3: PL spectra of the B-BIP in solutions and neat film

535nm
1 .0 B -B IP
Electroluminescence (arb. units)

0 .8

0 .6

0 .4

0 .2

0 .0

200 300 400 500 600 700 800

W a v e le n g th (n m )

Fig 4: EL spectrum of OLED fabricated in this work.

Figure 5. shows the energy level diagram of the 3.4eV and the optical band gap estimated from
HOMO and LUMO of the different organic the absorption onset, we can determine the
materials and the work function of cathode and HOMO energy at 6.5eV. In Comparison with
anode. By using cyclic voltammetry (CV) the energy level of B-BIP and NPB, B-BIP has
method obtaining LUMO energy of B-BIP is much higher hole injection barrier than that of

80
Marsland Press
Journal of American Science 2009: 5(1), 77-82
___________________________________________________________________________________
NPB. As a matter of fact, it is impossible for the
hole injection from ITO into B-BIP without the
assistance of NPB or some HTLs. This
diagram pointed out Alq3 has lower electron 4. CONCLUSION
injection barrier than that of B-BIP. Therefore, A new compound of emitter for OLED,
the electron injection from the MgAg into B-BIP BPh2(2-(benzimidazol-2-yl)pyridinato) (B-BIP),
will be enhanced and confines the recombination has been successfully synthesized and
zone at the interface between NPB and B-BIP. investigated. It has been shown that the novel
Fig.6 shows the current-voltage and ligands BIP is capable of chelating to B(III)
luminance-voltage characteristics of this device centers and the resulting compounds possess
having a low turn on voltage of about 4.5V for appreciable photoluminescent efficiency and
current and luminance. This device shows a very high thermal stabilities. This study further
brightness of 50 cdm-2 at the driving voltage of indicates that the emission band of the devices
12V with current density of 390 mA/cm2, could be modified by changing the composition
decaying to 25 cdm-2 in 100 hours. of emitting layer and therefore, OLEDs with
different colors could be obtained.

2.5eV

3.1eV
3.4eV 3.7eV
MgAg
NPB
Alq3
4.7eV
B-BIP
ITO 5.5eV
5.8eV

6.5eV

Fig 5: Energy level diagram of OLED materials, ITO, and Mg-Ag alloy

I-V
0.04 50
L-V

40
Luminance (cd/m )
Current density (A/cm )

0.03
2

30

0.02
20

0.01 10

0
0.00
-10
0 2 4 6 8 10 12
Voltage (V)

Fig 6: Current-voltage and luminance- voltage characteristics of OLED fabricated in this work

81
Synthesis, characterization and Electroluminescence of BPh2(2-(benzimidazol-2-yl) pyridinato) compound
Anchi Yeh
___________________________________________________________________________________
ACKNOWLEDGEMENTS Organic Electroluminescence,
Wissenschaft-und-Technik-Verlag, Berlin,
We wish to thank the National Science Council
(1996) 385.
of the Republic of China for financial support. [9] S. F. Liu, C. Seward, H. Aziz, N. X. Hu, Z.
Popovic, S. Wang, Organnometallics, 19
(2000) 5709.
REFERENCE [10] H. Schmidbaur, J. Lettenbauer, D. L.
[1] D. Song, S. F. Liu, R. Y. Wang, S. Wang, J. Wilkinson, G. Muller, O. Z. Kumberger,
Organomet. Chem. 631 (2001) 175. Naturforsch, 46B (1991) 901.
[2] Q. Liu, M. S. Mudadu, H. Schmider, R. [11] Anchi Yeh, Shen Cherng, Hsien-Chiao
Thummel, Y. Tao, S. Wang, Teng, The Journal of American Science,
Organometallics 21 (2002) 4743. 3 (4) 2007, 93-94.
[3] J. E. Lee, G. C. Choi, B. O. Rim, S. M. [12] Shen Cherng, Anchi Yeh, Hsien-Chiao
Kim, N. G. Park, Y. K. Ha, Y. S. Kim, Teng, The Journal of American Science,
Materials Sciencd and Engineering C 24 4 (1) 2007, 95-98.
(2004) 269. [13] R. Balamurugan, M. Palaniandavar, Inorg.
[4] C. W. Tang, S.A. VanSlyke, Appl. Ohys. Chem., 40 (2001) 2246.
Lett., 51 (1987) 913. [14] Z. K. Chen, H. Meng, Y. H. Lai, W.
[5] C. Adachi. S. Tokito, J. Tsutusi, S. Saito, Huang, Marcromolecules, 32 (1999)
Jpn. J. Appl. Phys., 27 (1988) 713. 4351.
[6] J. H. Burroughes, D. D. C. Bradley, A. R. [15] S. Tokito, H. Tanaka, K. Noda, A. Okada,
Brown, R. N. Marks, K. Mackay, R. H. Y. Taga, Appl. Phys. Lett., 70 (1997)
Friend, P. L. Burns, A. B. Homes. Nature, 1929.
347 (1990) 539. [16] Kim, J. H.; Matsuoka, M.; Fukunishi, K,
[7] J. R. Sheats, H. Antoniadis, M. Hueschen, Dyes and Pigments 40(1998) 53.
W. Leonard, J. Miller, R. Moon, D. [17] F. Peral, E. Gallego, J. Mol. Struct., 372
Roitman, A. Stocking, Science, 273 (1996) (1995) 101.
884. [18] A. Sarkar, S. Chakravorti, J.
[8] H. Nakada, T. Tohma. Inorganic and Luminescence, 65 (1995) 163.

82
Marsland Press
Journal of American Science 2009: 5(1), 83-87
___________________________________________________________________________________
Research Article
White Organic Electroluminescence Base on a new Aluminum
Complex

1
Anchi Yeh, 2Hsien-Chiao Teng
1
Department of Chemical and materials Engineering,
Chengshiu University, Kaohsiung, Taiwan, R.O.C
2
Department of Electrical Engineering
ROC Military Academy, Fengshan, Kaohsiung, Taiwan, R.O.C
E-mail: acyeh@csu.edu.tw ; scteng@nail,cma.edu.tw
______________________________________________________________
ABSTRACT: A bright blue emission material, tris{2-(benzimidazol-2-yl) pyridinato}
Aluminum (AlBIP) used for white-light of organic light emitting devices has been synthesized.
The decomposition temperature was observed at 400 к and no melting transitionȐTmȑwas
observed up to 400к. For three-layer LED devices with the configuration of ITO/NPB/AlBIP
/Alq3/MgAg, the white light emission covering the whole visible region from 400 to 700 nm
with the maximum brightness of 75 cd/m2 and current density of 330 mA/cm2 was observed.
[Journal of American Science 2009: 5(1), 83-87] (ISSN: 1545-1003)
Keywords: Electroluminescence; white light; device

1. INTRODUCTION one such as 8-hydroxyquinoline. Therefore,

White organic light emitting diodes have the thermal stability, an important character for
attracted much attention, because their potential the practical application in the electronic fields,
applications in the backlights of laptop of this metal complex is investigated by
computers and portable panel light sources. In thermogravimetric analysis (TGA) and
the literatures, several strategies including differential scanning calorimetry (DSC). The
multi-layer devices have been developed to organic emitting device using AlBIP as emitting
realize highly efficient white organic layer has been fabricated to study the
electroluminescence [1-5]. Luminescent chelate electroluminescent property of this metal
complexes have been shown to be particularly complex.
useful in electroluminescnt (EL) displays
because of their relatively high stability and 2. EXPERIMENTAL METHOD
volatility. The most well-known example of such The synthesis of the title compound was
chelate compounds is A1q3, not only a good accomplished by following processes, as shown
emitter but also a highly efficient in Scheme 1. The triethylaluminum solution
electron-transporting material, where q is the (25и w/w in hexane 1.86ml, 2.82×10-3 mole)
8-hydroxyquinolinato ligand [6, 7]. Via the was slowly added to 100 ml of THF solution
modification of the ligand of metal chelate containing 2-(2-pyridyl)benzimidazole (1.75g,
compound, the emission color of a metal chelate 9.0×10-3 mole) at 0oC under N2. After the
compound may be tuned. Other properties, resulting mixture was stirred at room
such as thermal stability and carrier mobility, temperature for 6 hours, 5 ml isopropyl alcohol
may also be improved upon. In the present was added to quench the reaction. The solvents
work, we report the synthesis and were removed under vacuum condition at 5×10-3
electroluminescent (EL) property of Torr, and the residual solid was sublimed to
tris{2-(benzimidazol-2-yl) pyridinato} purify the final product. Light green of AlBIP
Aluminum (AlBIP). The AlBIP containing was obtained in 85% yield. The formula of this
N,N-bidentate ligand instead of N,O-bidentate compound has been determined by 1H NMR and

83
White Organic Electroluminescence Base on a new Aluminum Complex
Anchi Yeh et al.
___________________________________________________________________________________
elemental analysis. The organic light emitting hole-transport material (HTM), and tris
device, Fig. 1, using AlBIP as the emitting and (8-quinnolinolato) aluminum (Alq3) was
electron-transporting layer were fabricated on employed as the electron-transporting material
the transparent conductive indium-tin oxide (ETM). The EL spectrum and the Commission
(ITO) glass substrate. The organic layers and Internationale de l’Eclairage (CIE) co-ordinates
the cathode were sequentially deposited by were measured by Pro-650 Spectroscanner (step
conventional vacuum vapor deposition in the size is 1.0 nm and bandpass is 4nm), the
same chamber without breaking the vacuum current-voltage (I-V) characteristic was
-6
under 3×10 Torr. The cathode composed of measured by Keithley 2400 Source meter.
magnesium silver alloy (Mg:Ag = 10:1) were Thermogravimetric analysis (TGA) was
deposited onto the top layer of organic materials performed on a Perkin-Elmer thermogravimeter
by co-evaporation of Mg and Ag from different (Pyris 1) under a dry nitrogen gas flow at the
source. Before the deposition, all of the heating rate of 20oC/min. Glass transition
organic materials were purified by the train temperature (Tg) and melting point (Tm) of
sublimation method. In the present wok, the materials were determined by differential
N,N’-bis-(1-naphthyl)-N,N’-diphenyl-1,1’-biphe scanning calorimetry of the Perkin-Elmer
nyl-4,4’-diamine (NPB) was used as the differential scanning calorimeter (DSC-7).

N
N
N Al(C2H5)3 N
N N
N
Al
HN N
N
N
N

Scheme 1. Synthesis process for the AlBIP complex.

Mg/Ag(200nm)

Alq3(50nm)
AlBIP(50nm)

NPB(50nm)
ITO

Substrate

Fig 1: The organic light emitting device (OLED).

84
Marsland Press
Journal of American Science 2009: 5(1), 83-87
___________________________________________________________________________________

3. RESULTS AND DISCUSSION observed in the spectrum of the AlBIP neat

The TGA of AlBIP that possesses a film. To investigate the electroluminescent
maximum rate of weight loss occurring at 400 properties of AlBIP typical three-layer device
o
C and no weight loss was observed at the with the configuration of
temperature lower than 350 oC. Above 600 oC, ITO/NPB/AlBIP/Alq3/MgAg was fabricated
there is about 14 wt % of residue composed of by using NPB as the hole-transporting layer and
aluminum ash. This aluminum complex is AlBIP as the emitter and Alq3 is
reasonably stable upon exposure to air and electron-transporting layer. The EL spectrum of
exhibited a high thermal stability in nitrogen. organic light emitting device at the bias voltage
The melting temperature (Tm) of AlBIP was not of 10 V, Fig. 3, shows the broader emission
observed up to 400 oC with DSC curve. The bands ranging from 400 to 700 nm were
DSC and TGA results indicate that the AlBIP observed, indicating that the three-layer LED
possesses a high thermal stability, which may device emitted white light covering the whole
serve as an advantage for the fabrication of visible light region. The band around 465 nm in
organic light emitting device because the use of EL spectrum can be attributed to emission of
the materials with high thermal stability as the AlBIP, because its emission position is almost
active emissive layer or carrier transporting layer identical with that in PL spectrum of AlBIP.
may provide the device with greater longevity The emission band at 525 is Alq3 emission
[11, 12]. position and 565nm can be attributed to the
The Photoluminescent (PL) spectra of the exciplex emission originated from the interface
AlBIP solutions and neat film, excited with 350 between NPB and AlBIP . The emission is
nm laser line, were illustrated in Figure 2. At almost fixed in the white region in the CIE
low concentration, 1×10-5 M in DMF, only one coordinate of x = 0.32 y = 0.37. For the small
emission band is observed with maximum at molecular organic materials, to develop the
450 nm, corresponding to the relaxation of double layer of device with white emission is
AlBIP from the excited state of a single very important because this kind of material is
molecule into ground state. There is red shift very seldom prepared so far, and it is very
emission band that a maximum at 460 nm is important for the fabrication of display panels.

-3
1x10 M
-5
1x10 M
neat film
1.0

0.8
photoluminescence

0.6

0.4

0.2

0.0

350 400 450 500 550 600 650

Wavelength(nm)

Fig 2: Photoluminescent spectra of the AlBIP in solutions and neat film

85
White Organic Electroluminescence Base on a new Aluminum Complex
Anchi Yeh et al.
___________________________________________________________________________________
The change of the spectral wavelength may 14]. At the same time important role here may
be achieved also by general conception of play electron-vibration interactions determining
search and design of modified materials for the spectral broadening of the emission lines.
wide band emission consists in substitution of So the future strategy of the materials design
the backside groups by electron acceptors like may be in this way also.
halogens etc. and different kind of donors [13,

Electroluminescence(arb.units) 0.10

0.08

0.06

0.04

0.02

0.00

400 500 600 700

W avelength(nm)

Fig 3: EL spectrum of OLED fabricated in this work.

Figure 4 shows the energy level diagram of diagram pointed out Alq3 has lower electron
the HOMO and LUMO of the different organic injection barrier than that of AlBIP. Therefore,
materials and the work function of cathode and the electron injection from the MgAg into AlBIP
anode. By using cyclic voltammetry (CV) will be enhanced and confines the recombination
method obtaining LUMO energy of AlBIP is zone at the interface between NPB and AlBIP.
2.8eV and the optical band gap estimated from Fig.5 shows the current-voltage and
the absorption onset, we can determine the luminance-voltage characteristics of this device
HOMO energy at 5.6eV. In Comparison with having a low turn on voltage of about 4.0V for
the energy level of AlBIP and NPB, AlBIP has current and luminance. This device shows a
much higher hole injection barrier than that of brightness of 75 cdm-2 at the driving voltage of
NPB. As a matter of fact, it is impossible for the 16V with current density of 330 mA/cm2,
hole injection from ITO into AlBIP without the decaying to 30 cdm-2 in 120 hours.
assistance of NPB or some HTLs. This

2.5eV
2.8eV
3.1eV
3.7eV
MgAg
NPB
AlBIP
Alq3
4.7eV
ITO
5.5eV 5.8eV
5.6eV
Fig 4: Energy level diagram of OLED materials, ITO, and Mg-Ag alloy

86
Marsland Press
Journal of American Science 2009: 5(1), 83-87
___________________________________________________________________________________

L-V and I-V characteristics of Al-BIP device

400 80

L-V
I-V

Current density(mA/cm )
2
300 60

Luminescence(cd/m )
40
200

20
100

2
0
0

0 2 4 6 8 10 12 14 16 18
Voltage(V)

Fig 5: Current-voltage and luminance- voltage characteristics of OLED fabricated in this work.

4. CONCLUSION [4] J. R. Sheats, H. Antoniadis, M. Hueschen, W.

A novel bright blue emission material, Leonard, J. Miller, R. Moon, D. Roitman, A.
tris{2-(benzimidazol-2-yl) pyridinato} Stocking, Science, 273 (1996) 884.
Aluminum (AlBIP), was successfully prepared [5] H. Nakada, T. Tohma. Inorganic and Organic
by the reaction of 2-(2-pyridyl)benzimidazole Electroluminescence,
and trisethylaluminum. Because of its high Wissenschaft-und-Technik-Verlag, Berlin,
thermal stability and excellent electrical (1996) 385.
characteristics, AlBIP and its related compound [6] S. –F. Liu, C. Seward, H. Aziz, N. –X. Hu, Z.
suggest a possible application for the use of Popovic, S. Wang, Organnometallics, 19
white-light of the organic light emitting devices. (2000) 5709.
[7] H. Schmidbaur, J. Lettenbauer, D. L.
Acknowledgements Wilkinson, G. Muller, O. Z. Kumberger,
This research was supported by the National Naturforsch, 46B (1991) 901.
Science Council of the Republic of China (Grant [8] T. R. Chen, A. C. Yeh and J. D. Chen,
No. NSC94 2113-M-230-001) Tetrahedron Lett., 46 (2005) 1569.
Corresponding Authors: [9] S. -F. Liu, Q. Wu, H. L. Schmider, H. Aziz,
Anchi Yeh N. –X. Hu, Z. Popovic, S. Wang, J. Am.
Hsien-Chiao Teng Chem. Soc., 122 (2000) 3672.
E-mail: acyeh@csu.edu.tw [10] Q.Wu, M. Esteghamatian, N. –X. Hu, Z. D.
E-mail: scteng@nail,cma.edu.tw Popovic, G. Enright, S. R. Breeze, S. Wang,
Angew. Chem. Int. Ed., 38 (1999).
Reference: [11] Z. –K. Chen, H. Meng, Y. –H. Lai, W.
[1] C. W. Tang, S.A. VanSlyke, Appl. Ohys. Lett., Huang, Marcromolecules, 32 (1999) 4351.
51 (1987) 913. [12] S. Tokito, H. Tanaka, K. Noda, A. Okada, Y.
[2] C. Adachi. S. Tokito, J. Tsutusi, S. Saito, Jpn. Taga, Appl. Phys. Lett., 70 (1997) 1929.
J. Appl. Phys., 27 (1988) 713. [13] M. Makowska-Janusik, J. Sanetra, H.
[3] J. H. Burroughes, D. D. C. Bradley, A. R. Palmers, D. Bogdal, E. Gondek, I. V. Kityk,
Brown, R. N. Marks, K. Mackay, R. H. Materials Letters, 58 (2004) 555.
Friend, P. L. Burns, A. B. Homes. Nature, [14] Albert J. van Reenen, Lon J. Mathias,
347 (1990) 539. Liezel Coetzee, Polymer 45 (2004) 79

87
 Journal of American Science, 5 (1), 2009 ISSN 1545-1003

  -RXUQDORI$PHULFDQ6FLHQFH

,661

January 10, 2009

The international academic journal, “The Journal of American Science” (ISSN: 1545-1003), is
registered in the United States, and invites you to publish your papers.

Any valuable papers that describe natural phenomena and existence or any reports that convey
scientific research and pursuit are welcome, including both natural and social sciences. Papers
submitted could be reviews, objective descriptions, research reports, opinions/debates, news,
letters, and other types of writings that are nature and science related.

This journal will be no charge for the manuscript contributors. If the author needs hard copy of
the journal, it will be charged for US$60/issue to cover the printing and mailing fee. Here is a
new avenue to publish your outstanding reports and ideas. Please also help spread this to your
colleagues and friends and invite them to contribute papers to the journal. Let's work together to
disseminate our research results and our opinions.

Papers in all fields are welcome, including articles of natural science and social science.

Please send your manuscript to editor@americanscience.org;

americansciencej@gmail.com

For more information, please visit http://www.americanscience.org;

http://www.sciencepub.org

Marsland Press
2158 Butternut Drive
Okemos, MI 48864
The United States
Telephone: (517) 349-2362
Email: editor@americanscience.org
americansciencej@gmail.com
Website: http://www.americanscience.org
http://www.sciencepub.org

Marsland Press, 2158 Butternut Drive, Okemos, MI 48864, The United States, (517) 349-2362
http://www.sciencepub.org iii editor@sciencepub.net
 The Journal of American Science





,661

Marsland Press
2158 Butternut Drive
Okemos, Michigan 48864
The United States
Telephone: (517) 349-2362

Emails: editor@americanscience.org;
americansciencej@gmail.com;
sciencepub@gmail.com

Websites: http://www.americanscience.org;
http://www.sciencepub.net;
http://www.sciencepub.org

ISSN 1545-1003

http://www.americanscience.org editor@americanscience.org

View publication stats