Chapter 2

Related Studies and Literatures

This chapter contains information about dengue, zika, chikungunya, Hagonoy

(Chromolaena odorata Linn.), Alagaw (Premna odorata Blanco) and mosquito repellent

effect and the relevant local and foreign literature and studies regarding the topic.

Local Literatures

Mosquitoes cause more human suffering than any other organism - over one

million people worldwide die from mosquito-borne diseases every year, according to

Nathan Cabello of The Philippine Star. Not only can mosquitoes carry diseases that

afflict humans, they also transmit several diseases and parasites that dogs and horses

are very susceptible to. In addition, mosquito bites can cause severe skin irritation

through an allergic reaction from the mosquito's saliva - this is what causes the red

bump and itching. Mosquito vectored diseases include protozoan diseases, i.e., malaria,

filarial diseases such as doing heartworm, and viruses such as dengue, encephalitis

and yellow fever. (The Philippine Star, 2015)

Dengue

Dengue was once just an intermittent part of the landscape, tropical Philippines

being hospitable to the mosquito that spreads the virus. Now and then, stories spike of

not only children but also grown men and women being felled by the disease, and

hospitals filled to overflowing with patients. But dengue is now a clear and present

danger all year round—no longer ―a rainy season disease,‖ according to the Department

of Health. In a report, the DOH’s epidemiology bureau said the number of dengue cases

in the country has risen a startling 35.7 percent, from 42,026 in Jan. 1-June 25, 2015,
 16

compared to 57,026 cases during the same period this year. That’s a lot of lives

endangered by dengue, and the death toll reflects this: 148 dead in 2015 and already

248 this year. In fact, the World Health Organization says dengue is now the world’s

most dominant mosquito-borne viral disease. And global warming due to climate

change is partly to blame. First identified in the 1950s, dengue is spread by the female

Aedes aegypti mosquito; the first documented cases were in the Philippines and

Thailand. The disease has since struck worldwide, with over 100 countries at risk and

390 million infections yearly. It takes 4-10 days to become obvious, with a high fever

and flu-like symptoms. (But that’s exactly what it’s not. It’s important not to mistake it for

the common flu.) It does not have a cure. The only thing to be done is to maintain the

patient’s bodily fluids until the disease runs its course. It’s crucial that dengue be

detected early, as the proper medical attention can lower mortality from 20 percent to

just less than 1 percent. Left unchecked or identified, dengue kills. It also spreads

quickly, as with any disease with a vector spread by mosquitoes. This is something

Filipinos know too well; we’ve been living with dengue for so long it now seems

ordinary. But ordinary it’s not; it’s dangerous. It’s virtually everywhere. The regions

currently hardest hit are Calabarzon (7,463 cases), Central Visayas (5,783), Central

Luzon (5,586), Northern Mindanao (5,521 cases) and Soccsksargen (4,583). Eastern

Visayas has the undesirable distinction of being the region with the biggest spike in

cases, a whopping 206-percent increase from 610 last year to 1,870 this year. Davao

has a 138-percent rise, and Central Visayas, a 137-percent increase. To think there is

now a dengue vaccine. It does not guarantee against contracting the disease, but helps

limit the risk of hospitalization by 80 percent and lower its life-threatening aspects.
 17

Developed by the French pharmaceutical firm Sanofi, Dengvaxia was approved for use

last December by the Food and Drug Administration, and the Philippines became the

first country to use it. (Press Reader, 2016)

The government and the private sectors are still implementing different measures

to combat dengue here in the Philippines as stated on the article in the Manila Bulletin

by Eduardo Gonzales (July, 2016) .The dengue immunization program is the latest one,

but still, despite of such programs, the incidence of dengue are continue to rise.

Department of Health (DOH) says that for the first half of 2016, there have been over

50,000 dengue cases nationwide about more than 15,000 cases in excess of what was

recorded over the same period last year. Also, our country is now experiencing a longer

rainy season which is peak period for dengue. It can contribute to the increasing cases

therefore the reported trend will continue as season can be considered as a factor. The

dengue vaccine (Dengvaxia) was just launched last April 4, 2016 and it is too early to

feel the changes that this vaccine might brought. As of June 2016, only a total of

300,087 students have been vaccinated. So we cannot expect that this vaccine could

eliminate dengue in the whole country. Additionally, this vaccine does not confer 100

percent protection it only protects two-thirds of those who receive it, although among

those who develop the illness after being immunized, it reduces the severe form of the

disease by ninth three percent. Also, the vaccine is not to be given to the entire

population only to those who are in the 9 to 45 years age bracket. Despite of those said

limitations, the University of the Philippines National Institute of Health stated that this

vaccine is expected to reduce dengue cases in our country by 24 percent in the span of

five years. Dengue is generally mild, self-limiting febrile illness that is sometimes
 18

complicate and gives rise to internal bleeding that is often fatal. It is caused by flavivirus

which is transmitted to human by a vector. Aedes aegypti is considered as the main

vector in the Philippines. It is known for having white dots on its back and head regions

and white stripes on its legs. Only female mosquito bites, as the blood is needed for

proper development of its egg. The mosquito prefers to bite during the day but it also

bites in the evening. It acquires the dengue virus when it bites and feeds on the blood of

a person with dengue. The virus then proliferates within the mosquito and eight to 11

days later, the mosquito becomes infective and can transmit the virus to any human it

bites. The female Aedes mosquito, which can live for 15 to 65 days and which has a

flight range of up to 300 meters, breeds in stagnant water. It lays up to a hundred eggs

at a time, in every place where non-running water exists: flower vases, jars, pots,

bottles, drums, roof gutters, drains, old tires, tree cavities, plant stumps, etc. The eggs

hatch into larvae (wrigglers) in a week, which become adult mosquitoes in another one

to one and a half weeks. Incidentally, if the pool dries up before the eggs hatch, the

eggs stay viable for up to a year and promptly hatch when the rains come.

Chikungunya

Chikungunya is a viral infection caused by the CHIK virus belonging to the

Togaviridae family. The virus is transmitted through the bite of infected daytime biting

female – primarily Aedes aegypti and Aedes albopictus – mosquitoes. They usually bite

during the early morning and late afternoon, and are typically found in and around urban

and suburban dwellings. Some mosquitoes that transmit Chikungunya in Africa also live

in forested areas. Monkeys and other wild animals are also believed to be reservoirs or

carriers of the virus. (Country Health advice-Philippines)

A town in Cavite province was declared under state of calamity due to

Chikungunya, as reported in CNN Philippines (Bonalos, 2016). Chikungunya outbreak

was declared after town of Indang recorded 99% of the total suspect Chikungunya

cases in Cavite. As of September 24, Indang reported 470 suspect cases. Chikungunya

is a mosquito-borne disease just like Dengue and Zika. While it shares the same

symptoms as Dengue — such as fever, joint pain and rashes — it is not fatal according

to the Cavite provincial epidemiologist, Nelson Soriano. The whole residents was

informed by the local health officials for continue to lower the peak of cases in the

province. (Bonalos, 2016)

Zika Virus

Zika Virus is transmitted from the bite of an infected mosquito called Aedes

aegypti, which is also responsible for other mosquito-borne diseases such as Dengue

fever and Chikungunya. Aedes mosquitoes are usually active during daytime, but they

can be also active at night. The virus can also be transmitted via unprotected sexual

intercourse. Pregnant women are among the most those who are in most danger when

infected by the disease, since it can be passed to the fetus, and has been linked to a

neurologic defect called microcephaly. Microcephaly is a congenital condition where the

baby is born with an undersized head, which results in incomplete brain development

and death. Unfortunately, Zika can't be treated since there is no available vaccine for it

yet. The first case of Zika virus in the Philippines was confirmed last August, 2016. A

45-year-old married woman from Iloilo City marked the first case of Zika virus in the

Philippines confirmed by Philippines' Department of Health Secretary Paulyn Jean B.

Rosell Ubial. The woman was positive with both strains of virus. Upon arrival at the local
 20

hospital in Western Visayas on August 31, urine and blood samples that were collected

from the victim tested positive for Zika virus. The patient complained of skin rash and

joint pains, which are common symptoms of Zika virus, but did not have a fever.

Contrary to rumors, she is not pregnant, which saves her from the possibility of having a

baby with birth defects like microcephaly (which is commonly linked to Zika). DOH

suspects that the virus was transmitted to the patient since she had no history of

travelling to Zika affected countries in the past two weeks. On September 30, the United

States Embassy in the Philippines released a bulletin informing US citizens that the

Centers for Disease Control and Prevention (CDC) had issued Travel Guidance for the

Philippines regarding the risk of Zika virus infection. "Zika virus is endemic. The risk of

Zika to travelers, especially pregnant women, in endemic countries is likely lower (but

not zero) than in areas where Zika is newly introduced and spreading widely," the

bulletin read. Women who are pregnant or planning to become pregnant should discuss

their travel plans with their medical provider and consider postponing nonessential

travel to the Philippines. The World Health Organization classified the Philippines under

Category 2, which refers to countries with possible endemic transmission or evidence of

local mosquito-borne Zika infections in 2016. Zika, which resembles a light case of the

flu, is transmitted by mosquito species found in tropical and sub-tropical regions: Aedes

aegypti and Aedes albopictus, or tiger mosquitoes. The disease is strongly suspected of

causing birth defects such as microcephaly and other brain deformities in newborns.

The Philippines is known for its natural resources, being a tropical country, its

supply of naturally occurring medicinal plants can be observed everywhere. Thus, the

practice of traditional medicine has been passed on from generation to generation. Its
 21

significance has further deepened in the health care delivery considering the expensive

western medicine which most Filipinos cannot afford. (Dotado-Maderazo, 2010).

Insect Repellent

Insect repellents are important tools for prevention of insect-borne diseases as

well as painful or uncomfortable insect bites. The 2 most effective and widely used

products are N, N-diethyl-3-methylbenzamide (DEET), which is applied to exposed skin,

and permethrin, which is applied to clothes. Both are safe when used according to

directions. Other repellents, including a variety of plant-derived products, have also

been used but generally have a weaker or shorter-lived repellent effect. Insect

repellents are important public health tools for the prevention of vector-borne infectious

diseases. Technically, an insect repellent is any chemical -- natural or synthetic -- that

causes insects or other arthropods to make directed, oriented movements away from

the source of repellent. Chemical substances that have repellent effects enable us to go

places and do things in insect- and disease-infested areas while avoiding disease

transmission by arthropods. ―Insect repellents should be used to avoid mosquito bites,‖

Health Secretary Paulyn Ubial said as she called on the public to include eliminating

mosquito breeding sites. Health experts noted that Aedes aegypti mosquitoes carry a

triple threat to public health: dengue, Chikungunya and Zika viruses. (Jaymalin, 2016)

Alagaw

The Plant sample, Premna odorata Blanco (―Alagaw‖ in Filipino) from the family

of Verbenaceae/Lamiaceae genus Premna, common names are Alagau (Tagalog),

Abgau (Bisaya), Fragrant premna (English), Saliargao (Central Bisaya) is a small hairy

tree 3 to 8 meters high. Leaves are ovate to broadly ovate, 10 to 20 centimeters long,
 22

with broad, rounded, or somewhat heart-shaped base, and pointed tips. Under surface

of the blade is usually covered with short hairs, aromatic when crushed. Flowers are

greenish-white or nearly white, 4 to 5 millimeters long and borne on terminal

inflorescences (cymes) 8 to 20 centimeters in diameter. Fruit is fleshy, dark purple,

rounded, about 5 millimeters in diameter. Found in the secondary forests in the

Philippines abundant in a low-altitude thicket. The water decoction of the leaves is used

to treat patients with tuberculosis problems, vaginal wash or douche, flatulence (gas

pain) fever, colds, cough, bronchitis and fever blisters of the lips. In addition, the

decoction of shoots used as parasiticide. (Stuart, 2016) It is also utilized locally to

manage phlegm, stomachache and headache. The leaves of Premna odorata Blanco

are also used to treat wounds by direct application to the infected area. (Quisumbing,

1978)

Previous literature utilized P. odorata Blanco leaves ethanolic crude extract but

did not show cytotoxic activity against HeLa cells using micro titration cytotoxicity assay.

This is in contrast to the results obtained in the study showing toxicity to other cell lines.

P. odorata hexane extract exhibited excellent cytotoxic activity against all cancer cells

lines used in the study and this is the first report for the biological activity of Premna

odorata bark. (Jacinto, 2015)

Hagonoy

Chromolaena odorata Linn. Is from the family of Asteraceae, genus

Chromolaena, common names are Christmas bush, Common floss flower, Devil weed,

Siam weed (English) is a bushy herb or subshrub with long rambling branches,

spreading into tangled dense thickets up to 2 meters high. Base of the plant is hard and
 23

woody while the branch tips are soft and green. Leaves are arrow head-shaped, 5 to 12

centimeters long and 3 to 7 centimeters wide, with three characteristic veins in a

pitchfork pattern, growing in opposite pairs along stems and branches. Flowers are in

clusters of 10 to 15, tubular, pale pink-mauve or white, 10 millimeters long, at the end of

the branches. Seeds are dark, 4 to 5 millimeters, narrow and oblong, with a parachute

of white hairs which turn brown as the seeds dry with aqueous and methanolic extracts

of leaves yielded carbohydrates (1.10%), protein (24.08%), lipid (14%), fiber (50%), ash

(10.98%), with an energy content of 220.20 kcal. Leaves were also a rich source of

mineral elements such as Ca, Na, K, Fe, Mn, Zn, Cu, P, and Mg. Leaves also yielded

alkaloids, flavonoids, saponins, cyanogenic glycosides, tannins, and phytic acid. In the

Philippines, Hagonoy crushed leaves used for "kulbebra," boils and tumorous

inflammatory conditions while the concoction of juices of leaves and fruits of hagonoy is

used for skin diseases and boils. It also used to stop bleeding and wound healing while

young leaves are used to treat wounds. (Stuart, 2016)

Local Studies

Another study for new alarming disease was conducted in our country. In March

2012, a prospective longitudinal cohort study, which includes active surveillance for

acute febrile illness, was initiated in Cebu City, Philippines (Emerging infectious

diseases, 2015, Vol. 21). Participants contacted study staff to report fever and were

also contacted weekly by staff to determine if they had fever during the previous 7 days.

Fever episodes triggered an acute-illness visit by a study nurse, who performed a

clinical assessment of the patient and collected an acute-phase blood sample. During

the first year of surveillance, 270 acute febrile illnesses were detected; 267 of the
 24

patients had samples available for serologic testing for evidence of Influenza, Dengue,

Chikungunya, Japanese encephalitis, and Zika virus infections. In May 2012, a 15-year-

old boy in Cebu City reported a subjective fever, an acute-illness investigation followed.

Other symptoms included headache, conjunctivitis, sore throat, myalgias, stomach pain,

anorexia, nausea, and vomiting, but no rash. The boy did not seek medical care or

require hospitalization; his only treatment was acetaminophen. He had no recent travel

history, and no other members of his household were ill. The boy recovered fully by the

3-week study follow-up visit. An acute-phase blood sample, collected 2 days after

symptom onset, was negative for dengue and Chikungunya viruses by reverse

transcription PCR. An in-house dengue/Japanese encephalitis IgM/ IgG capture ELISA

and Chikungunya ELISA were used to test paired acute- and convalescent-phase blood

samples; all results were negative. ZIKV ELISA was not available at the testing

laboratory. However, by using real-time reverse transcription PCR targeting the gene

that encodes the precursor of membrane protein, they detected ZIKV RNA in the

patient’s serum sample. Serologic evidence of ZIKV infection has been reported in

Thailand, Vietnam, Malaysia, Indonesia, and the Philippines.

In the study about the Anti-immunosuppressive Effects of Chromolaena odorata,

Leaf Extract in Cyclophosphamide-injected Balb/C Mice held in University of the

Philippines,Diliman, QuezonCity assessed the Immunomodulatory effects of C. odorata

in vivo. The results showed that the plant crude extract could potentially reverse the

inhibitory effects of cyclophosphamide in the various immune parameters measured in

this study. These could provide baseline information in considering the efficacy of the

plant extract as immuno-nutritional supplement and in this study could also suggest that
 25

there could be another antimicrobial mechanism, aside from ROS production, that is

induced by C. odorata extracts. The decrease in the production of reactive oxygen

species could also indicate the possible antioxidant property of C. odorata plant

extract.(Leonora P. Nudo, June 2012)

Pure and ethanolic crude extracts of leaves of hagonoy (Chromolaena odorata

L.) were tested for its phytochemical constituents and were found to contain alkaloids

and saponins. Chromolaena Odorata L. has the medicinal components and drug

potentials for manufacture of pain killers,tranquilizers and anti-biotics. The anti-bleeding

bioactivity of hagonoy was tested on five male mice and findings proved that hagonoy

has also anti-bleed components and could treat and dress wounds and could stop

bleeding. The greater the volume of hagonoy was applied on the wound of the male

mice, the faster the bleeding stopped. The study proved that the pure extract of

hagonoy was an anti-bleed as it was able to stop bleeding of the wound of five male

mice, thus, hagonoy could be used to treat and dress wounds. (Jeon, 2014)

The present study was conducted to test for in vivo Brine Shrimp Lethality Assay

(BSLA) of the ethanolic extracts of Lantana camara, Chromolaena odorata, and

Euphorbia hirta and correlate cytotoxicity results with known pharmacological activities

of the plants. Novel cytotoxic, antitumor, and pesticidal compounds can be isolated from

potential plant sources through the assessment of cytotoxic activity against brine

shrimps. Cytotoxicity was evaluated in terms of LC50 (lethality concentration). Ten

nauplii were added into three replicates of each concentration of the plant extract. After

24 hours the surviving brine shrimp larvae were counted and LC50 was assessed.

Results showed that the extracts of L. camara, C. odorata, and E. hirta were potent
 26

against the brine shrimp with LC50 values of 55, 10, and 100 ppm (μg/mL), respectively.

It indicated that bioactive components are present in these plants that could be

accounted for its pharmacological effects. Thus, the results support the uses of these

plant species in traditional medicine. (Lilybeth F. Olowa, 2013)

According to (Allan Patrick Macabeo, April 2014) the results demonstrate that the

medicinal plant Premna odorata possesses antimycobacterial compounds, and support

the purported traditional use of this plant in the treatment of TB. In the first report of the

presence of compound 1 in Premna odorata have its antimycobacterial activity. In the

study also reports on the absence of antimycobacterial activity of compound 4, and the

weak activity of the sterol mixture of compounds 2 and 3 as compared to the pure

compounds. Based on the results, it is strongly believed that the leaves of Premna

odorata represent a promising source of antimycobacterial agents that merits further

investigation.

Based on the study of (Javier, 2010) the insecticidal action of water andorganic

solvent extracts of 10 plants – makabuhai, ―luyang dilaw‖ (Curcuma longa L.), lagundi

(Vitexnegundo) ―lagunding dagat‖ (Vitextrifolia), wild sunflower, derris (Derris elliptica)

"kakawate‖ (Gliricidia sepium), ―hagonoy‖ (Chromolaena odorata), ―alagaw-gubat‖

(Premna auseosa) and ―alagaw‖ (Premna odorata Blanco.) were evaluated against

brown planthopper (Nilaparva talugensl), green leafhopper (Nephothettix species),

diamondback moth (DBM) (Plutella xylostella L.) and Asian corn borer(Ostrinia

furnacalis). Water extracts of ―makabuhai‖ showed systemic and ovicidal toxicities and

growth inhibitory effect (IGR). ―Lagundi‖ and wild sunflower showed contact and

ovicidal toxicities and IGR effect. Derris sp. and ―luyang dilaw‖ showed contact and
 27

ovicidal toxicities (Morallo-Rejesus et. al. 1993). There were numerous plants that repel

mosquitoes including citronella or lemon grass (for its geraniol and citronellol

constituents), horsemint also known as bee balm orlemon bee balm (works the same

way as the citronella grass because of its strong incense like, peppermint scented

aroma that confuses the mosquitoes by masking the smell of its usual host),marigolds

(which contains pyrethrum that is also used in insect repellents), ageratum also known

as white weed or floss Flowers (which emits an offensive smell for mosquitoes because

of itsc oumarin content used in commercial mosquito repellents), and catnip which

contains nepetalactone that act as an irritant to mosquitoes (R. T. Tampican, 2012)

Foreign Literatures

Mosquitoes are one of the deadliest insect in the world. Its ability to carry and

spread diseases to more than 700 million people every year that can mostly lead to

death is quite startling. According to latest WHO report, As of June 30, 2016 there are

1,792 cases affected by Chikungunya in BuhaHawa region in Kenya. Over 80% of their

population and 50% of health work force are infected by the said disease. In 2015, more

than 3.2 billion people were at risk of having mosquito related diseases and the ongoing

Malaria transmission was found in 95 countries and territories. In Sub-Saharan Africa

there is a higher share of global malarial burden with 86% of cases and 90% global

deaths. Recently, there are cases reported in Florida, USA and Yunan in China of

Dengue. At present, the most advisable prevention against this is to apply a protectant

that can drive away mosquitoes (WHO 2015).

Plants and Constituents with Mosquito Repellency Properties

Human beings have used plant parts, products and secondary metabolites of

plant origin in pest control since early historical times. Vector control has been practiced

since the early 20th century. During the pre-DDT or Dichlrodiphenyltrichloroethane era,

reduction of vector mosquitoes mainly depended on environmental management of

breeding habitats, i.e. source reduction. During that period, some botanical insecticides

used in different countries were Chrysanthemum, Pyrethrum, Derris, Quassia, Nicotine,

Hellebore, Anabasine, Azadirachtin, d-limonene camphor, Turpentine (Rahumanet al.

2008).From the early 1950’s, DDT and other synthetic organochloride and

organophosphate insecticides were extensively used to interrupt transmission of vector

borne diseases by reducing densities, human-vector contact and, in particular, the

longevity of vector mosquitoes. In the mid-1970s, the resurgence of vector borne

diseases, along with development of insecticide resistance in vector population,

because to poor human acceptance of indoor house spraying and environmental

concerns against the use of insecticides led to a rethinking in vector control strategies

(WHO 2005).

Controlling Mosquitoes

Application of alternative methods in mosquito control as part of the Integrated

Mosquito Management (IMM) has been gaining importance (Ghosh et. al.2012). IMM is

a decision-making process for the management of mosquito populations, involving a

combination of methods and strategies for long-term maintenance of low levels of

vectors. The purpose of IMM is to protect public health from diseases transmitted by

mosquitoes, maintain healthy environment through proper use and disposal of

pesticides and improve the overall quality of life through practical and effective pest
 29

control strategies. The main approaches of IMM include: (a) Source reduction and

habitat management by proper sanitation, water management in temporary and

permanent water bodies, and channel irrigation. Vegetation management is also

necessary to eliminate protection and food for mosquito larvae; (b) Larviciding by

application of dipteran specific bacteria, insect growth regulators, surface films and oils,

expanded polystyrene beads, phytochemicals, organophosphates and organochlorides,

by application of synthetic pyrethroids, organophosphates and synthetic or plant

derivedrepellents, insecticide impregnated bed nets, genetic manipulations of vector

species, etc.;(d) Use of mosquito density assessment in adult and larval condition and

diseasesurveillance; and (e) Application of biological control methods by

usingentomophagous bacteria, fungi, microsporidians, predators and parasites. Of

theabove avenues of IMM, larviciding approach is the more proactive, pro-environment,

target specific and safer approach than controlling adult mosquitoes. Vector control

offers a viable alternative to reduce the spread of vector borne diseases (Chowdhuri

etal. 2009).

Prevention to Minimizing the Incidence of Mosquito Borne Disease

In addition to personal protection and educating the public, the most successful

method of minimizing the incidence of mosquito-borne diseases is to eradicate and

control the mosquito vectors, which is performed principally by systematic treatment of

the breeding places through a combination of environmental management and

application of larvicides that do not harm other organisms in the environment (Cetin et

al. 2004; Corbel et al. 2004). Organophosphorus, carbamate, and pyrethroid

insecticides are less persistent, as they break down quickly in theenvironment, and are,
 30

therefore, recommended as classical larvicides. Nevertheless, pyrethroids are very toxic

to fish and should not be used where there are fish orcrustaceans (Rozendaal 1997).

The most commonly used larvicides are the organophosphorus compounds such as

temephos, fenthion, and chlorpyrifos, whichare highly active against mosquito larvae

and other aquatic insects. Temephos is the larvicide of choice for Aedes aegypti and

Anopheles control, while fenthion and chlorpyrifos are used against Culex species. (Das

et al. 2007). Questions have been raise to whether these larvicides would cause

environmental pollution and hazards to human health and other non-target organisms if

recurrently applied. Repeated use of synthetic insecticides for mosquito control has

disrupted natural biological control systems and led to resurgences in mosquito

populations (Das et al. 2007). At present, the most successful measures to decrease

the incidence of mosquito borne diseasesare by personal protection and control of the

vectors.

DEET as Mosquito Repellent

In 1953, DEET (N, N – Diethy-m-Toluamide) were discovered. It is a member of

the toluene chemical Family and much known effective mosquito repellent. A study

released showed that DEET based repellants aren’t just hazardous to mosquito

however highly toxic because of toluene property. An organic solvent used in rubber

and plastic cements. DEET is absorbed through the skin and passes into the blood.

One of the effect of DEET in the body are hard to breath, have cough, minimal

alertness, seizures, trouble stomach and vomiting. Proper care should be observe when

using the product. From the human health point of view, mosquito bites are more
 31

nuisance than serious health threat, choosing a botanical based repellant makes more

sense (Paterson et. al. 2001).

Plants Constituents to Control Mosquitoes

The use of plant derived compounds for mosquito control has been reported

since 1933. Plants used in traditional medicine recorded in ethno pharmacological

literature provided a source of information for such investigation. Medicinal plants with

larvicidal properties have paramount importance for the local control of mosquito

(Debella et. al. 2007). A large number of plant-derived substances possess

physiological and behavioral activities against insect pests and may provide new source

of natural pesticides. Natural products have shown that it is possible to produce a great

range of biological activities, including toxicity, repellent action, antifeedant and growth

regulation properties (Huang and Ho 1998; Chiam et al. 1999). Plant terpenoids have

dominated the subject of chemical ecology since they have been studied for their

activities against a variety of insect models (Gonzalez-Coloma et. al. 1995). The great

ecological interest of the plants terpenoids has been reflected in several studies, finding

strong toxicant effects that produce larval mortality, significant growth inhibition,

repellent, and fumigant activity.

Chemicals Use to Kill Insect

The presence of so-called secondary compounds, which have no known function

in photosynthesis, growth or other aspects of plant physiology, give plant materials or

their extracts their anti-insect activity. Secondary compounds include alkaloids,

terpenoids, phenolics, flavonoids, chromenes and other minor chemicals. These

chemicals may kill, retard or accelerate development or interfere with the lifecycle of the
 32

insect in other ways (Bell et al. 1990). So, these chemicals can disrupt major metabolic

pathways and cause rapid death, act as attractants, deterrents, phagostimulants, and

anti-feedants or modify oviposition. Although compared with modern synthetics the plant

substances are relatively weak. The botanical insecticides are generally pest-specific,

readily biodegradable and usually lack toxicity to higher animals (Bowers 1992).

Factors affecting the Efficacy of the Phytochemicals

The efficacy of phytochemicals against mosquito larvae can vary significantly

depending on plant species, plant parts used, age of plant parts (young, mature or

senescent), solvent used during extraction as well as upon the available vector

species(Shaalanet al. 2005). (Sukumaret al. 1991) have described the existence of

variations in the level of effectiveness of phytochemical compounds on target mosquito

species vis-à-vis plant parts from which these were extracted, responses in species and

their developmental stages against the specified extract, solvent of extraction,

geographical origin of the plant, photosensitivity of some of the compounds in the

extract, effect on growth and reproduction. Changes in the larvicidal efficacy of the plant

extracts can occur due to geographical origin of the plant response in the different

mosquito species and due to variation in the species of plant examined and between

plants parts used to study the larvicidal efficacy (Kishore et al. 2011). However, the

principal objective of the present study is to report the changes in larvicidal potentiality

of the plant extracts due to change of the particular solvent used during extraction.

Foreign Studies

According to T.C. Kazembe et al. (2012), the mosquito repellent properties of

Capsicum frutescens, Carica papaya and Cyanodondactylon extract and extract mixture
 33

has a big impact in insect-borne diseases control. The result showed that C. frutescens

and C. papaya were effective for 2.5 hours. The C. dactylon was effective only for 1.5

hours. While the mixtures of C. frutescens and C. papaya last longer for 4 hours by

repellence test. Analysis of Variance (ANOVA) showed that there 5% significant

difference among three.

Volatile oils extracted by steam distillation from four species plants Curcuma

conga, Citrus hystrix, Cymbopogon winterianus, Ocimum americamum.According to

Tawatsin etal. (2001),their repellency effects against three mosquitoes vectors, Aedes

aegypti, Anopheles dirus, and Cutex quinquefasciatus were can be replace the DEET

and co-formulated with vanillin to prolong the effect.

According to S. Nkomo et al. (2007) the active constituents of Blumea alata,

Biden spilosa and Chenopodium ambrosiodes were extracted by dry distillation where

oil were separated from aqueous phases. The result showed effective mosquitoes

repellent.

Based on experimental study of J.R. Eifert,Cymbopogon citrulus may be used as

a partial replacement for DEET based repellents. Cymbopogon citrulus was found not to

be as effective compared into the 15% DEET compound many people are too sensitive

to the DEET chemical to use it as a repellent.

Study of citronella leaf based on herbal mosquito repellent using natural binders

such as neem powder, potatostarch, corn starch, coconut shell powder, wood powder,

and cow dung. An attempt has been made to prepare 100% herbal product based on

citronella leaf remains which is left out and of no use after steam distillation. Efficacy of

prepared citronella leaf cakes were evaluated on three different parameters such as
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flammability, burning time, and mosquito repellency test. The cakes were also sprayed

with different concentration of citronella oil. Based on result, the residual percentage of

combination of cakes was calculated and it suggested the neem powder has most

effective among the group with 10% impregnated citronella oil (N. Rani et. al. 2013).

A preliminary screening was carried out of five plants: Azardirachta indica, Citrus

medica, Murraya koenigi, Ocinum tenuifloreum, Ricinus communis for repellent activity

against mosquitoes. It was studied under natural condition making use of traditional

knowledge as background. Observation undertaken in houses wherein dried powder

leaves (250g) burnt produce smoke act as repellent. Result indicated that among five

plants Azardirachta indica, Murraya koenigi, and Citrus medica provided repellence for

6 hours followed by Ricinus communis and Ocinum tenuifloreum with four hours. Aside

from these the incidence of mosquito bites significantly reduced after the usage of plant

products (B. Sai Shankar et. al. 2013).

According to Yimer et al. (2014) the Artemisia annua with eucalyptus oil

combination has better performance than neem oil and rose oil. It showed up to 2 hours

protection against female mosquitoes.

Oil of Mentha piperita L., a widely used essential oil was evaluated for larvicidal

activity against different mosquito species Aedes aegypti, Anopheles stephensi, Cutex

quinquefasciates by exposing in 3rd larvae of mosquito in enamel trays 6x4 inch size

filled to depth of 3 inch with water among three species tested Cutex quinquefasciates,

was most susceptible followed by Ae. aegypti and An. stephensi. The emergence at

3ml/m was also inhibited to a great extent and the few adults which emerged did not

oviposit even after taking blood meal. The oil showed strong repellent action against
 35

adult mosquito when applied on human skin. Percent protections obtain against An.

annuralis, An. calicifacies, and Cx. quinquefasciates was 100%, 92.3% and 84.5%

respectively. The repellent action of Mentha piperita L. oil was comparable to Mylol oil

consisting of dibutyl and dimetyl phthalates (M.A. Ansari, et. al. 1999).

Essential oil from 12 plants medicinal plants were evaluated by different bioassay

methods (vaporizer,filter,aerosol) for knock-down and adulticidal efficacy on filarial

vector Cutex quinquefasciates. Five essential oils were identified as potential treatment

calamus, camphor, citronella, clove, eucalyptus. The result showed that the knock-down

time decreases with increases concentration in clove oil treatment. The cinnamon oil

was effective which recorded as median lethal time. Camphor were found less toxic by

aerosol method. The result suggested that clove oil and cinnamon oil have potential to

use as eco-friendly approach for control of the major important filarial vector under

laboratory condition (M. Ramar et. al. 2014).

The repellent and growth-inhibiting effects of turmeric oil (Curcuma longa),

sweetflag oil (Acoruscalamus), neem oil (Azadirachtaindica) and Margosan-O (a

commercial neem-based insecticide) on red flour beetles (Tenebrionidae) are also

studied. It was found that the repellency increased with increasing concentration of the

oils and Margosan-O, while the turmeric oil or sweet flag oil repelled insects during the

first 2 weeks, while some essential oils have been used asinsect repellents since

ancient ages such as citronella and pennyroyal. Repellents of plant origin do not pose

hazards of toxicity to human and domestic animals and are easily biodegradable.

Natural products are safe for human when compared to that of synthetic compounds

(Sharma et al. 1993a; Sharma and Ansari 1994).

The repellent properties of plants to mosquitoes and other pest insects were well

known before the advent of synthetic chemicals. In southern India, leaves of Vitex

negundo are burned to repel mosquitoes from houses (Curtis et al. 1989). The plant

products have been used traditionally to repel or kill the mosquitoes in many parts of the

world (Novak 1985). Forty essential oils extracted from Australian plants were evaluated

against mosquitoes, march flies, and sand flies. The most effective of these were

Dacrydium franklini, Backhousia myrtifolia, Melaleuca bracteata, and Zieria smithii

(Penfold and Morrison 1952). Repellency properties of Nepetalactone

(cyclopentanoidmonoterpene) isolated from the catnip plant, Nepetacataria, against 17

species of insects was reported by Eisner (1964). Also, many monoterpenes were

reported for their insect repellent properties such as α-pinene, limonene,

terpinolene,citronellol, citronellal, camphor, rotundial, dolichodial, teucrein, and

isoborneol (Takikawaet al. 1998; Eisner et al. 2000). Among 29 tested alkaloids

obtained from Delphinium, Consolida, and Aconitum species, 21 compounds showed a

promising insect repellent activity, while eight of them were not active. Hetisine had the

highest activity while venulol showed the lowest (Ulubelenet al. 2001).

Rajkumar and Jebanesan (2009) have tested that the oviposition deterrence of

Cassia obtusifolia leaf extract against An. stephensi. Mehra and Haridar (2002) also

described ovi position deterrence of Cuscuta hyaline. Rajkumar and Jebanesan (2005b)

also tested the leaf extract of Solanum trilobatum against the gravid females of An.

stephensi. Melia azedarach leaf extract is also reported to have oviposition and

deterrence against A. aegypti (Coria et al. 2008), (Autranet et al. 2009) has reported

that the essential oil from leaves and stems of Piper marginatum exhibited anoviposition
 37

deterrent effect against A. aegypti. (Elangoet et al. 2009) reported that the oviposition

deterrence of acetone, ethyl acetate and methanol extracts of Aegle marmelos,

Andrographis lineate and Cocculus hirsutus.

Results from a study by (Perichet et al.1995) suggest that adulticidal action is a

potential and unexploited field of research. The seed kernel of Azadirachta indica, the

leaf of Cymbopogon nardus and the wood of Fernandoa adenophylla were also

reported to have adulticidal properties against Aedes aegypti (Zaridahet al. 2006).

Above 90% mortality was found in the ethyl acetate and methanol extract of all

experimental plants at the concentrations of 500μg/mL (Kamaraj and Rahuman 2010).

The adult mortality was found in methanol extract of Artemisia nilagirica, with the LC50

and LC90 values of 205.78 and 459.51ppm for An. stephensi, and 242.52 and 523.73

ppm for A. aegypti, respectively (Panneerselvam et al. 2012).