Insecticidal Activity of Kantutai (Lantana camara) and Atis (Annona

squamosa) (Annona squamosa) Leaves Against Mealybugs (Pseudococcidae

sternorrhyncha)

Aquino Z., Ariscon J., Cabrera R., Cabuguang A., De Guzman H., Gatche J., Guiao M.,

Indico M., Ramirez J., and Ramos J.

Malabon National High School – M. Naval St., Hulong Duhat, Malabon City

ABSTRACT

This study utilized the insecticidal property of Kantutai (Lantana camara) and Atis (Annona

squamosa) leaves against mealybugs (Pseudococcidae sternorrhyncha). The plants used in this

study were environmental friendly and does not cause any respiratory diseases. The bio-

insecticide was made from the ethanolic extract of Kantutai (Lantana camara) and Atis (Annona

squamosa) leaves. This study aims to produce a mealybug insecticide out of environmental

friendly materials. There are three concentrations each for the plant extract and for the combined

extracts. Kantutai (Lantana camara) leaves concentrations: A1 (0.25g/500mL), A2

(0.375g/500mL), and A3 (0.50g/500mL). Atis (Annona squamosa) leaves concentrations: B1

(0.25g/500mL), B2 (0.375g/500mL), and B3 (0.50g/500mL). Kantutai (Lantana camara) and

atis (Annona squamosa) leaves concentrations: C1 (0.25g/500mL), C2 (0.375g/500mL), and C3

(0.50g/500mL). Four tests were conducted; survival test, morphological analysis, time to

eliminate, and phytochemical analysis. The researchers inferred that both Kantutai (Lamara

camara) and Atis (Annona squamosa) leaves are qualified as an alternative insecticide to

eliminate mealybugs (Pseudococcidae sternorrhynca).

Keywords: Survival Test, Morpological Analysis, Time to Eliminate, and Phytochemical

Analysis

INTRODUCTION

These days, pesticides have components that can adversely affect the atmosphere,

contaminate bodies of water, and reduce the quality of crops. It is caused by the chemicals

contained in certain pesticides; Thus, the use of pesticides is harmful to the environment, and it

contributes to pollution. In spite of this, it is still being used because of the unending breeding

and emerging of insects.

An insect called mealybug (Pseudococcidae sternorrhyncha), a plant-sucking insect, this

insect causes leaf curling, and yellowing as it weakens the plant accompanying by its excretion

of honeydew as it feeds that results of stickiness of the plant and enhance the chance of the

growth of sooty molds (Walton and Pringle 2009). These pests are greatly found in warm

growing climate, the damage is insignificant at early infestation but in higher pest level they are

already spreading throughout the plant leaves to roots; Hence, controlling the pests are often not

complete as some of the population are under the bark or cordon and can hide from their natural

predators. Forcing the use of chemical pesticides which can affect non-target species.

This study aims to determine the efficacy of Kantutai (Lantana camara) and Atis

(Annona squamosa) leaves as organic insecticide against mealybugs (Pseudococcidae

sternorrhyncha) and would have better effect than a commercial insecticide. In this study, it will

promote the use of tropical plants on the Philippines and make an eco-friendly insecticide.
 Using alternative insecticide with the use of Kantutai (Lantana camara) and Atis

(Annona squamosa) leaves ethanolic extract, farmers, or local residence can utilize those and

make insecticide out of it. Considering that it is an insecticide for mealybugs (Pseudococcidae

sternorrhyncha), it can be used in greenhouses safely thinking that it would not affect the plants'

health and the soil mineral content.

Specifically, this study will utilize Kantutai (Lantana camara) and Atis (Annona

squamosa) leaves ethanolic extract as a bio-insecticide. It subsumes the determination of the

insecticidal activity of different concentration of the extract to mealybugs (Pseudococcidae

sternorrhyncha).

METHODOLOGY

Research Design

Table 1. Proportion of Components in Each Concentration

Concentrations Kantutai Leaves Atis Leaves

Amount of Amount of Amount of Amount of

solvent (mL) solute (g) solvent (mL) solute (g)

A1 500 0.25 - -

A2 500 0.375 - -

A3 500 0.50 - -
B1 - - 500 0.25

B2 - - 500 0.375

B3 - - 500 0.50
C1 500 0.25 500 0.25

C2 500 0.375 500 0.375

C3 500 0.50 500 0.50

Gathering of Materials

Kantutai (Lantana camara) leaves were accumulated at Malabon National High school

Garden. Atis (Annona squamosa) leaves were collected in the researchers’ residential area. The

researchers gathered exact 1kg of fresh Kantutai (Lantana camara) and Atis (Annona squamosa)

leaves with the total of 2kg of leaves. The leaves were cleaned and rinsed with distilled water to

remove impurities and dirt. Five liters of 95% technical alcohol were used. The mealybugs

(Pseudococcidae sternorrhyncha) were gathered at the residential area of the researchers.

Drying and Pulverizing of the Kantutai (Lantana camara) and Atis (Annona squamosa)

Leaves

The researchers sun dried the plant leaves, then used the hot air oven of Malabon

National High school Laboratory for the 1kg of Kantutai (Lantana camara) and 1kg Atis

(Annona squamosa) fresh leaves. The leaves were put in the oven for 30 minutes in 30 o Celsius.

After drying, the air-dried leaves were pulverized using an electric blender. The ground sample

were kept in a ziplock bag.

Preparation of the Ethanolic Extract of Kantutai (Lantana camara) and Atis (Annona

squamosa) Leaves
 The total mass of four hundred grams of pulverized Kantutai (Lantana camara) and six-

hundred grams of pulverized Atis (Annona squamosa) dried leaves were measured using O-

HAUS digital scale. It was placed in the 1000ml Erlenmeyer flask. Pulverized samples were

soaked with one liter of 95% technical grade ethyl alcohol. The flask was covered for 24 hours.

After the infusion, filtration process was done using a filter paper. Resulting filtrates were

concentrated in the rotary evaporator for 2-5 hours at estimately 40ºC. The solvent from the

extract were removed using rotary vacuum evaporator to collect the crude extract. The extraction

was done at the DOST in Quezon City, Philippines.

Phytochemical Analysis of Kantutai (Lantana camara) and Atis (Annona squamosa) Leaves

Two test tubes containing 10ml of the ethanolic extract were labeled Kantutai (Lantana

camara) and Atis (Annona squamosa) leaves extract. 12 M Ammonium hydroxide was added to

the extract for analysis. The mixture was bathed in warm water for 15-60 minutes. The change of

color was observed and recorded. There was a gradual development of a strong red or violet

color and it indicated the presence of flavonoids in the extract. Bate-Smith method was used for

analysis. The procedure was conducted in Malabon National High School.

Preparation of Treatment

Eleven bottles were prepared, two for the controlled variables and nine for the

experimental variables. Atis (Annona squamosa) had three different concentrations; A1, A2, and

A3, the distilled water had a contant volume of one liter, but varied amount of the crude extract

weighing 0.25ml, 0.375ml, and 0.5ml respectively. Kantutai (Lantana camara) also had three

different concentrations; B1, B2, and B3, the distilled water had a constant volume of 500ml, but

varied amount of the crude extract weighing 0.25ml, 0.375ml, and 0.5ml respectively. Lastly,
combined crude extract of Atis (Annona squamosa) and Kantutai (Lantana camara) had three

different concentrations; C1, C2, and C3, the distilled water had a contant volume of one liter,

but varied amount of the crude extract weighing 0.25ml, 0.375ml, and 0.5ml respectively.

Rearing of Mealybugs

The mealybugs (Pseudococcidae sternorrhyncha) were kept in a holed large sterile

container. The mealybugs were cultivated using a pumpkin as food source with proper humidity

and were kept at room temperature ranging from 23ºC to 28ºC (Roltsch 2006). The cultivation

was conducted on the laboratory of De La Salle – Canlubang, Laguna that reproduced enough

insects for the experiment.

Testing

The experimental group which is the alternative insecticide were tested to determine its

efficacy. The investigation was reported from an average of three (3) replicates. Researchers

used Survival test to evaluate the effectiveness of the crude extract compared to the control

groups, and Morphological test to determine the mutilated parts of the mealybugs

(Pseudococcidae sternorrhyncha) using a digital microscope (Cooltech).

Survival Test

The test was conducted into the 33 petri dishes containing one infested plant part with 10

mealybugs (Pseudococcidae sternorrhyncha) each. Twenty-seven for the concentrated crude

extract, three for the commercial insecticide which is the positive control and other three for the

distilled water which is the negative control. The concentrated extract was placed in spray

bottles, one each per concentration. Three replicates were made for each treatment. The alive

mealybug was counted with the interval of two minutes, the percentage of survival was reported
inferring to three different concentration of the extract comparing it to the commercial

insecticide and to the distilled water.

The survival rate of mealybugs (Pseudococcidae sternorrhyncha) of each concentration

was computed using the formula:

Percentage
Survival (%) = 100
Base

Where:Percentage = total number of alive mealybugs in each concentration

Base = total number of mealybugs used

The mortalities of treated groups were computed according to Abbott's formula:

X-Y
Mortality (%) = X 100

Where: X = percentage survival in the untreated control and Y = percentage survival in the

treated sample.

Morphological Analysis of Knockdown Mealybug

The morphological changes of the treated mealybugs were studied, recorded, and further

compared to the controlled mealybugs for each concentration of the extract after 2 hours. The

digital microscopy was performed. The abnormalities of the mealybugs were studied with the use

of the digital microscope (Cooltech) and pictures will be taken.

Time to Eliminate

To determine which concentration exterminate mealybugs the fastest will be compared to

the positive control, the length of extermination time was measured accordingly to 30 replicates.
The researchers record the time it took in eliminating 5 mealybugs per concentration. Each

concentration contained 10 mealybugs. Using a timer, knockdown mealybugs was recorded.

Statistical Treatment

The following statistical tools were used in this study.

Mean. This was used in determining the average extermination and surviving larvae for

the Survival Test. The mean was obtained be adding the results per concentration and dividing it

by the number of trials. The mean was also usedd for obtaining the average time of

exterminating per concentration.

ANOVA (One Way Analysis of Variance). This was used to test the significant

difference of plant extracts from the positive control in Survival Test and Time to Eliminate. In

Survival Test, the measured significance was the efficacy of the plant extracts in contrast to the

positive control with the number of exterminated mealybuug. In Time to Eliminate, the

significance measured was the total period of extermination for both groups. The f-value was

equated to the tabular value on 0.05 level of confidence.

RESULTS

This study determined the efficacy of Kantutai (Lantana camara) and Atis (Annona

squamosa) leaves extract as an insecticide against mealybugs (Pseudococcidae sternorrhyncha)

as prepared with the following tables, graphs, and figures.

Phytochemical Analysis

Qualitative Test Results for Flavonoids in Kantutai and Atis.

Table 2. Phytochemical Present in the Extract of Kantutai and Atis

Plant Extract Results

Kantutai Leaves Extract Positive Change to strong red

Atis Leaves Extract Positive Change to dark violet

Survival Test

The researchers conducted the survival test in a set of three replicates per concentration.

Each replicate contained ten (10) mealybugs (Pseudococcidae sternorrhynca).

Table 3. Results of Survival Test

Concentration No.of alive mealybugs Mean Percentage Rank

Trial 1 Trial 2 Trial 3
A1 (0.25g:500ml) 4 3 5 4 40% 4
A2 3 4 2 3 30% 6

(0.375g:500ml)
A3 (0.50g:500ml) 2 0 1 1 10% 9
B1 (0.25g:500ml) 5 4 6 5 50% 2
B2 (0.375g:500ml) 3 4 2 3 30% 6
B3 (0.50g:500ml) 4 2 1 2.33 23.30% 7
C1 (0.25g:500ml) 5 5 4 4.67 46.70% 3
C2 (0.375g:500ml) 4 3 3 3.33 33.30% 5
C3 (0.50g:500ml) 2 2 0 1.33 13.30% 8
NCon 10 10 10 10 100% 1
PCon 0 0 0 0 0% 10
 120
Percentage Survival Rate of Mealybugs
100

80
Percentage of Survival Rate

0.25g/500ml
0.375g/500ml
60
0.50g/500ml
Negative Con.
Positive Con.
40

20

0
Kantutai Atis Kantutai and Atis Negative Con. Positive Con.

Graph 1. Summary of Results in Survival Test

Table 4. Results of Mortality Test

Concentration No.of eliminated Mean Percentage Rank

mealybugs
Trial 1 Trial 2 Trial 3
A1 (0.25g:500ml) 6 7 5 6 60% 7
A2 7 6 8 7 70% 5

(0.375g:500ml)
A3 (0.50g:500ml) 8 10 9 9 90% 2
B1 (0.25g:500ml) 5 6 4 5 50% 9
B2 (0.375g:500ml) 7 6 8 7 70% 5
B3 (0.50g:500ml) 6 8 9 7.66 76.6% 4
C1 (0.25g:500ml) 5 5 6 5.33 53.3% 8
C2 (0.375g:500ml) 6 7 7 6.66 66.6% 6
C3 (0.50g:500ml) 8 8 10 8.66 86.6% 3
NCON1 0 0 0 0 0% 10
NCON2 0 0 0 0 0% 10
NCON3 0 0 0 0 0% 10
PCON1 10 10 10 10 100% 1
PCON2 10 10 10 10 100% 1
PCON3 10 10 10 10 100% 1
 120

Percentage Mortality Rate of Mealybugs

100
Percentage of Survival Rate

80

0.25g/500ml
0.375g/500ml
60
0.50g/500ml
Negative Con.
Positive Con.

40

20

0
Kantutai Atis Kantutai and Atis Negative Con. Positive Con.

Graph 3. Summary of Results in Mortality Test

Percentage of Mortality Rate
Table 5. F- Test of Mortality Rate

SUMMARY
Averag Varian

Groups Count Sum e ce

5.86666 11.9809

Trial 1 15 88 7 5
13.0285

Trial 2 15 93 6.2 7
Trial 3 15 96 6.4 14.4

ANOVA
Source of

Variation SS df MS F P-value F crit

Between 2.17777 1.08888 0.92060 3.21994

Groups 8 2 9 0.08289 2 2
Within 551.733 13.1365

Groups 3 42 1

553.911

Total 1 44        
Time to Eliminate

Table 6. Time to Eliminate Mealybugs

Concentration Time to Total Mean Ranking

eliminate(minutes) (minutes)
A1 6 5 7 18 6 7
A2 3 4 3 10 3.33 3
A3 3 2 2 7 2.33 2
B1 6 6 5 17 5.67 6
B2 5 4 5 14 4.67 5
B3 4 3 3 10 3.33 3
C1 7 8 6 21 7 8
C2 4 4 4 12 4 4
C3 3 4 3 7 3.33 3
Negative Con. - - - - - 9
8
Positive Con. 1 1 1 3 1 1

6
Average Time to Eliminate Mealybugs
5

4 Average Time (Minutes)

0
Kantutai Atis Kantutai and Atis Negative Con. Positive Con.

0.25g/500ml 0.375g500ml 0.50g/500ml Negative Con. Positive Con.

Graph 4. Summary of Results in Time to Eliminate Mealybugs

Table 7. F- Test for Time to Eliminate

SUMMARY
Averag Varianc

Groups Count Sum e e

Time to 3.81818 4.56363

eliminate(minutes) 11 42 2 6
3.72727 5.01818

11 41 3 2
3.54545 4.47272

  11 39 5 7

ANOVA
Source of Variation SS df MS F P-value F crit
0.42424 0.21212 0.04527 0.95579 3.3158

Between Groups 2 2 1 8 7 3
140.545 4.68484

Within Groups 5 30 8

140.969

Total 7 32        
Morphological Analysis

Figure 3. Summary of Morpological Analysis

Concentration 1 (0.25) Concentration 2 (0.375) Concentration 3 (0.50)

Kantutai

(Lantana

camara)
This mealybug's body This mealybug's burnt

got distorted because of The leg of this mealybug body is enveloped with

the 0.25 Kantutai is crippled while the its sap in order to give
Description
(Lantana camara) whole body has a golden birth to its 3 younglings

concentrations. Its whole and brown tint. as a last ditch effort to

body has a golden tint. continue their bloodline.

 Atis (Annona

squamosa)

The mealybug that is The mealybug that was

submerged in Atis submerged in Atis

This mealybug curled its
(Annona Squamosa) (Annona Squamosa) with
body up with clear signs
Description with a concentration of a concentration of 0.5 is
of being burnt with a
0.25 is burnt and crippled so mutilated; its body got
golden tint.
with clear signs of burnt in half. Its upper

mutilation. body has a brownish tint.

Kantutai

(Lantana

camara) and

Atis (Annona

squamosa)
This mealybug's body is
This mealybug is so This mealybug's body is
crippled and distorted
mutilated that its body is burnt with a bloated
Description with clear signs of
extremely crippled with a upper body along with
mutilation along with a
dark brown tint. golden tint.
brownish tint.

Positive Control

The mealybug is crippled

up and its carcass is

 scorched and burnt up

with a dark brown tint.

Alive Mealybug

The untested mealybug's

''body'' has an oval shape

Description form, it has a snowy camo

that is visible all over its

hairy body.

DISCUSSIONS

Phytochemical Analysis

Both Kantutai (Lantana camara) leaves and Atis (Annona squamosa) leaves showed

positive result in the presence of flavonoids because of their change in color to vivid violet. In a

study conducted Anand J., et al. in Analysis of antioxidant activity, total phenolic content and

total flavonoid content of Lantana camara leaves and flowers showed the present phytochemical

investigations on Kantutai (Lantana camara) leaves extracts revealed the presence of flavonoids,

phenols, terpenoids, anthocyanins, carbohydrates, and proteins in the methanolic extracts of

Lantana leaves and flowers.

In the study conducted by Paul R., et al. in “Phytochemical Screening of Annona

squamosa and Haematological Studies in Clarias batrachus” showed the phytochemical

investigations revealed that Atis (Anona squamosa) leaves extracts are positive of the presence

of alkaloids, steroids, tannins, phenols, reducing sugar, saponin and flavonoids.

Survival Test

The survival rate of mealybugs (Pseudococcidae sternorrhynca) of each concentration

was computed. The higher the percentage of survival rate, the higher the number of mealybugs

(Pseudococcidae sternorrhynca) that survived. The Concentration C1 (0.25g/500ml) with a

percentage of 46.70% had the highest chance of mealybugs (Pseudococcidae sternorrhynca)

survival. While the Concentration A3 (0.50g/500ml) with a percentage of 10% had the lowest

chance of mealybugs (Pseudococcidae sternorrhynca. survival.

The different concentration of atis (Annona squamosa) leaves extracts showed similar

results to the percentage of survival rate of positive control.

Mortality Rate of Mealybugs

Results were observed with the quantity of knockdown mealybugs (Pseudococcidae

sternorrhyncha) in three trials in each concentration under a constant of ten minutes. The

mortality rate of each concentration was computed. Excluding the positive and negative control,

the mortality rates were compared. Concentration A3 (0.50g/500mL) produced the most

satisfactory result, ranked first in the mortality test with a rate of 90% and a mean of 9

knockdown mealybugs (Pseudococcidae sternorrhynca). The Concentration B1 (0.25g/500mL)

had the least number of knockdown mealybugs (Pseudococcidae sternorrhynca) with a mean of

5 and rate of 50%. The most favorable mortality rate was obtained from the highest

concentration of Kantutai (Lantana camara) while the lowest concentration of Atis (Annona

squamosa) was the least.

Statistical Treatment of Mortality Test

It presents the difference in the mortality rate of mealybugs (Pseudococcidae

sternorrhynca) for the experimantal over control. The computed F-value of 0.08 was clearly

lower than the tabular value of 3.22 at 0.05 level of confidence therefore, there is a significant

amount of difference in the mortality rate of mealybugs (Pseudococcidae sternorryncha) of the

control from the one which used plant extract. Thus, the results shown do not support the

experiments executed by the researchers.

Time to Eliminate

The test indicates the time the extracts took place to eliminate mealybugs

(Pseudococcidae sternorrhynca) for 3 trials. Each concentration involved 10 mealybugs

(Pseudococcidae sternorrhynca). Positive control ranked first with the fastest average time of 1

minute to eradicate all of the mealybugs (Pseudococcidae sternorrhynca). Concentration A3

ranked 2nd took 2 minutes and 33 seconds to eliminate all of the mealybugs (Pseudococcidae

sternorrhynca). Concentrations A2, C3, and B3 ranked 3rd to extinguish the mealybugs

(Pseudococcidae sternorrhynca). that has an average of 3 minutes and 33 seconds. Concentration

C2 ranked 4th with an average time of 4 minutes to exterminate the mealybugs. Concentration

B2 ranked 5th with an average time of 5 minutes 7 seconds. Concentration B1 ranked 6th with an

average time of 6 minutes and 7 seconds. Concentration A1 ranked 7th with an average time of 6

minutes. Concentration C1 ranked 8th with an average time of 7 minutes. While negative control
ranked 9th and did not eliminate all the mealybugs (Pseudococcidae sternorrhynca). Moreover,

the positive control eliminated most of the mealybugs (Pseudococcidae sternorrhynca) faster

than the experimental group.

Statistical Treatment of Time to Eliminate

It shows the difference in the time of extermination of mealybugs (Pseudococcidae

sternorryncha) for the experimental over the control. The computed F-value of 0.45 was clearly

lower than the tubular value of 3.32 and confidence level of 0.05 therefore, there is a clear

significant amount of difference in the time to exterminate mealybugs (Pseudococcidae

sternorryncha) of the control from the one which used plant extract. Thus, the results shown

subsides the experiments executed by the researchers.

Morphological Analysis

The mealybugs were viewed using the digital microscope. Abnormalities were observed. For the

experimental group the deformities in the mealybugs had been observe. The control group

showed some similar abnormal changes at the mealybugs body parts. The experimentation

justified that the mealybugs died due to the high concentrations of the plant extracts. Based on

the researchers’ observation the mealybugs' body was mutilated proving that it wasn't

exterminated by being drowned. Hence the mealybugs died due to the high toxicity of the plant

extract.
CONCLUSION

Based on the results, the researchers concluded that Kantutai (Lantana camara) and

(Annona squamosa) can be utilized as an alternative insecticide against mealybugs

(Pseudococcidae sternorrhynca).

RECOMMENDATION

The researchers want to recommend to the future researchers to use a higher

concentration and to widen their knowledge about plants and pest

ACKNOWLEDGEMENT

The completion of this study does not only need the cooperative effort of the researchers.

This study will not be possible without the support of experts and professionals. To the following

people who contributed to this study we would like to express our sincerest gratitude for their

effort to help in the success of this study.

Dr. Ma Victoria De Gulan, Principal of Malabon National High School for supporting

our whole research team in this Science Investigatory Project.

Mrs. Anacoreta R. Trogo, Head of the Science Department for her whole hearted support

in our Research team morally and financially.

Mrs. Mylene J. Aguinaldo, our project adviser with her full support, inspiration, and her

eagerness to accomplish our project.

Mrs. Marilou Duque, Grade 10 Science teacher, for lending us the laboratory apparatuses

we needed in this investigatory project.

 To the researchers’ friends and classmates for their support and encouragement that gave

them the strength to continue improving this project.

To all members of the researchers’ family for their outmost understanding, concern, and

love which provided them the inspiration to complete this work.

Lastly, and above all, to God Almighty, for his divine grace for the researchers to do all

that is needed to accomplish this investigatory project.

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doi:10.1673/031.009.4701

Electronic Sources

Morton J., “Sugar Apple” https://hort.purdue.edu/newcrop/morton/sugar_apple.html

Walton C., “Lantana camara” http://www.iucngisd.org/gisd/species.php?sc=56

“Mealybugs” https://www.planetnatural.com/pest-problem-solver/houseplant-pests/mealybugcon

trol

“Why organic: harmful effects of chemical pesticides” http://www.havahart.com/why-organic-

harmful-effects-of-chemical-pesticides
Appendix A

COLLECTION AND PREPARATION FOR THE PLANT EXTRACT

Collection of Kantutai and Atis Leaves

Oven-drying Pulverization of Kantutai and Atis Leaves

of Kantutai and Atis Leaves

Weighing of pulverized components

Appendix B
EXTRACTION OF KANTUTAI AND ATIS LEAVES

Grinding of Components Soaking of Components in 95% ethyl alcohol

Separation of Solvent from Kantutai and Atis Leaves Extract

Separated solvent from Kantutai and Atis Leaves Extract

APPENDIX C

Gathering of Mealybugs

Cultivation of Mealybugs
APPENDIX D

Preparation of plant extracts

Final concentrations of plant

extract

Preparation for the control

Appendix E
PHYTOCHEMICAL ANALYSIS
SURVIVAL TEST

Preparation for the Survival Test

During the Survival Test

TIME TO ELIMINATE

Preparation of Time to Eliminate Observation after application

MORPHOLOGICAL ANALYSIS

Preparation of the digital microscope

Magnification of mealybugs
APPENDIX F
COMPUTATION OF ANOVA IN MORTALITY TEST

Positiv
Negativ
e
A B C e
Contro
Control
l
Trial 1 3 4 3.67 3 0
Trial 2 2.33 3.33 3.33 3 0
Trial 3 2.67 3 2.33 3 0
Total 8 10.33 9.33 9 0
Mean 2.666667 3.443333 3.11 3 0

SUMMARY

Groups Count Sum Average Variance

Trial 1 15 88 5.866667 11.98095

15 93 6.2
Trial 2 13.02857

Trial 3 15 96 6.4 14.4

ANOVA

Source of
SS df MS F P-value F crit
Variation
Between
2.177778 2 1.088889 0.08289 0.920602 3.219942
Groups
Within
551.7333 42 13.13651
Groups

Total 553.9111 44
APPENDIX G
COMPUTATION OF ANOVA IN TIME TO ELIMINATE

Positiv
Negativ
e
A B C e
Contro
Control
l
Trial 1 7 6.33 6.33 0 3
Trial 2 7.67 6.67 6.67 0 3
Trial 3 7.33 7 7.67 0 3
Sum 22 20 20.67 0 9
Average 7.333333 6.666667 6.89 0 3

SUMMARY

Group Varianc
Count Sum Average
s e
Trial 1 11 42 3.818182 4.563636
Trial 2 11 41 3.727273 5.018182
Trial 3 11 39 3.545455 4.472727

ANOVA

Source of
SS df MS F P-value F crit
Variation
Between
0.424242 2 0.212121 0.045278 0.955797 3.31583
Groups
Within
140.5455 30 4.684848
Groups

Total 140.9697 32