Available online at www.sciencedirect.
com
ScienceDirect
Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188
AicQoL2014Kota Kinabalu
AMER International Conference on Quality of Life
The Pacific Sutera Hotel, Sutera Harbour, Kota Kinabalu, Sabah, Malaysia
4-5 January 2014
“Quality of Life in the Built & Natural Environment”
Biolarvacidal Potential of Ipomoea Cairica Extracts Against
Key Dengue Vectors
Ahmad Razali Ishak *, Nazri Che Dom, Hazilia Hussain, Nur Husna Sabri
Department of Environmental Health, Faculty of Health Sciences, Universiti Teknologi MARA
Abstract
Chemicals are used widely as insecticides causing adverse effects to human health and the environment. Natural
insecticides derived from plants which is eco-friendly. This study is to assess the efficacy of Ipomoea cairica extract
against two different mosquito larvae. Lethal Concentrations (LC50) of petal, leaves and root extract against Ae.
albopictus larvae were 20.5, 27.9 and 34.3 mg/L respectively. Lethal concentration (LC 50) were found to be lower in
Ae. aegypti which were 12.7, 13.6 and 31.9 mg/L respectively. In conclusion, these extracts can be utilized as an
option in vector control program.
©©2014
2014The
TheAuthors.
Authors.Published
Published byby
Elsevier Ltd.Ltd.
Elsevier ThisSelection
is an openand
access article under
peer-review theresponsibility
under CC BY-NC-ND of license
the Association of
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Malaysian Environment-Behaviour Researchers, AMER (ABRA Malaysia).
Peer-review under responsibility of the Association of Malaysian Environment-Behavior Researchers, AMER (ABRA malaysia).
Keywords: Ae. Aegypti; ae. albopictus; dengue; ipomoea cairica
1. Introduction
Mosquito population is a well-known vector in transmitting many types of diseases causing serious
public health problems (Gubler 1995; Lam, 1993; Nazri et al. 2013a). In Malaysia, there are about 434
species of mosquitoes representing 20 different genera (Zaridah et al. 2006). Many studies have indicated
that Ae. aegypti and Cx. quinquefasciatus are the most abundant urban mosquitoes found in Malaysia
(Raghavendran et al. 2011). The common method used currently in Malaysia to control mosquito’s larvae
is through the usage of chemicals such as abate. However, extensive use of chemical insecticides have
*Corresponding author. Tel.:+60-0332584509; fax: +60-0332584599
E-mail address:
[email protected]1877-0428 © 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Peer-review under responsibility of the Association of Malaysian Environment-Behavior Researchers, AMER (ABRA malaysia).
doi:10.1016/j.sbspro.2014.10.052
Ahmad Razali Ishak et al. / Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188 181
created problems in insecticide resistance indicates a reduction of sensitivity of a mosquito towards
insecticides developed due to the widespread usage of chemicals in destroying larvae. Thus, it is
considered to be a serious public health challenge that contributes to ineffective method for disease
control (Nazri et al. 2013b). Moreover, in ecological perspective, the widely used pesticide also kills non-
targeted predators for mosquitoes such as dragonfly and other insects. A study conducted by
Hidayatulfathi et al. (2005) revealed that insecticide resistance has developed in three different species of
mosquitoes (Cx. quinquefasciatus, Ae. albopictus and Ae aegypti) larvae against Malathion, Permethrin
and Temephos. The study also reported that Cx. quinquefasciatus developed higher resistance to
Malathion and Permethrin as compared to Ae. Albopictus and Ae. aegypti. There are known specific cure
has been found for most vector borne diseases, reducing the larval population is the best in controlling the
outbreak of dengue. Abu Bakar et al. (2009) tested on M. cajuputi plant extracts against dengue vector in
the laboratory setting. The result showed high mortality of 60.2% – 61.4% and 60.8% - 64.0 % in Ae.
aegypti and Ae. albopictus when sprayed with 5% and 10% M. cajaputi after exposure for 5 and 10
seconds respectively. Their study was extended using the same plant species in aerosol spray cans at low
cost housing flat in Kuala Lumpur and discovered that there was a high knockdown and mortality rate in
both Aedes species towards the MS aerosol (Abu Bakar et al. 2012).In addition, the repellent activities of
several plants were also tested against Aedes species in Malaysia. Misni et al. (2008) investigated the
repellency of Piper aduncum Linn essential oil against Ae. aegypti and their results indicated optimum
repellency (ED50 value 1.236 ugcm-2 and ED 900.8 ugcm-2) occurred after 90 seconds exposure. For the
effect against Ae. albopictus, the lowest median effective dose (ED50) value was 2.1 ug/cm2 at 90 seconds
of exposure (Misni et al. 2009). Furthermore, field evaluation by ultra-low-volume (ULV) spray against
dengue vector using Litsea elliptica extracts and bifenthrin indicated low larvacidal effect but the extract
caused 96.5 -97.6% mortality among Ae. aegypti adults (Sulaiman et al. 2006). Thus, these findings
exhibit the potential of using natural products in combating vector borne diseases. Other studies have also
shown the advantage of using plant extracts as larvacide because they do not induce pesticide resistance
in the mosquitoes (Azmathullah, 2011; Borah et al. 2010; Choochote et al. 2004; Chowdry et al. 2006
and Katade et al. 2006). Recently, natural products derived from plants extracts are widely used as an
alternative method against mosquitoes due to their excellent properties like low toxicity to human and a
high degree of biodegradability thus reducing the risk of adverse ecological effects. In view of residue
problems in the environment and the development of insect resistance to synthetic insecticides like DDT
and other chlorinated hydrocarbons, the recent trend is to explore the use of plants to obtain extracts that
are safe for non-targets animals and do not pose any residual problem. The working concept of this
research is to assess the ability of Ipomea cairica (morning glory) as a larvidcide and its efficacy to
control vector mosquito population. This study addressed the two main objectives in achieving full
understanding of this potential effect of natural product. (1) To screen the phytochemical properties of of
Ipomea cairica. (2) To investigate the efficacy of different part of morning glory extract (Ipomea cairica)
as larvacide to Ae. aegypti and Ae. Albopictus using larvacidal assay technique.
2. Material and Methods
2.1. Plant material extraction
The samples of Ipomea cairica plant were collected and washed using distilled water to avoid
possible trace contamination. Then, the different parts of Ipomoea cairica such as petals, roots and leaves
were dried at room temperature (25 - 28 °C) and grounded separately to produce fine particles. By using
modern soxhlet extraction technique, Soxterm, 10 gram of each powdered material was inserted into
soxterm extraction thimble. The samples were soaked in 500 ml of methanol in solvent vessel. The
182 Ahmad Razali Ishak et al. / Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188
soaked material were left for 3 hours in Soxterm extractor. The vessel containing the crude extract was
dried in the dryer at 85 °C about 1 hour. The weight of crude extract was calculated according to the
following formula:
Crude extract (gram) = (m2 - m1)/E (1)
The formula indicates m2 = weight of the dry empty vessel in grams, m1 = weight of the vessel in grams
containing crude extract after evaporation of the solvent and E = the sample weight in grams (10 gram).
The volume of stock solution should be 20 ml of 1%, obtained by weighing 200 mg of Ipomoea cairica
crude extract and adding 20 ml methanol to it (WHO, 2005). The crude extract was kept in a screw-
capvial, with aluminium foil over the mouth of the vial. These extractions were preserved in a refrigerator
at 4°C temperatures with proper labeling until it was used in phytochemical screening and mosquito
larvicidal bioassay tests.
2.2. Phytochemical screening
Qualitative determinations of phytochemical constituents were carried out on the methonolic
extracts of Ipomoea cairica as described by Prashant et al, 2011. Nine phytochemical constituent were
screened including saponins, alkaloid, phenol, tannin, glycoside, flavonoid, Amino acid, phytosterol and
terpen.Ferric Chloride Test chloride solution. Formation of bluish black color indicates the presence of
phenols. The extract was run through Gelatin’s Test for tannin screening. In this method, 1% gelatin
solution containing sodium chloride was added. Formation of white precipitate indicates the presence of
tannins. Detection of glycosides was carried out by modified Borntrager’s Test. The plant extract was
hydrolyzed with diluted hydrochloric acid. Then, the extract was treated with Ferric Chloride solution and
immersed in boiling water for about 5 minutes. The mixture was cooled and extracted with an equal
volume of benzene. The benzene layer was separated and then the solution was treated with ammonia
solution. Rose-pink color was formed in ammonical layer indicates the presence of anthranol glycosides.
For flavonoids screening, Alkaline Reagent Test was used. In this test, the extract was treated with few
drops of sodium hydroxide solution. Intense yellow color was formed, which becomes colorless on
addition of dilute acid indicates the presence of flavonoids. Froth test was used for saponins detection.
Plant extract was diluted with distilled water to 200 ml. This solution was then shaken in a graduated
cylinder for 15 minutes. 1 cm layer of foam formed which indicates the presence of saponins.
Xanthoproteic Test was used to detect the presence of protein and amino acids. The extract was treated
with few drops of concentration nitric acid. Yellow color formation indicates the presence of proteins.
Salkowski’s Test was used for phytosterols detection. The extract was treated with chloroform and filter.
The filtrate was treated with few drops of concentrated sulphuric acid, mixes well and allowed to stand.
Appearance of golden yellow color indicates the presence of triterpenes.The extract was dissolved in
water and treated with 3-4 drops of copper acetate solution. Emerald green color formation indicates the
presence of diterpenes.
2.3. Preparation of mosquito’s larvae
Ae. aegypti and Ae. albopictus were obtained as egg raft column on a filter paper from Vector
Control Research Unit (VCRU) University Science of Malaysia (USM) Penang. Larval food was added to
the distilled water 24 hours prior to the placement of the collected eggs to promote uniform hatching. The
Ahmad Razali Ishak et al. / Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188 183
hatched larvae were then transferred to shallow pans containing 2 liter of tap water. The containers were
kept at 25 + 2 °C throughout the process.
2.4. Preparation of mosquito’s larvae
Bioassays for the toxicity of the extracts were carried out against Ae. aegypti and Ae. Albopictus
larvae using World Health Organization (WHO) Standard Method 2005. In the larvicidal assay, two types
of mosquito larvae at the third instars stage were exposed to test concentration of 10, 25, 50, 75 and 100
pm of different part of Ipomoea cairica extract into the disposal test cups. Three replicates for
eachconcentration together with one control with distilled water were set up simultaneously. The tests
were conducted at room temperature at 25 to 27 oC. The knockdown rate was recorded every 5 minutes
within 1 hour after 24 hours the test larvae exposed to the concentration. Mortality was recorded each 4
hours until 48 hours exposure. During this period of time, no food was offered to the test organisms
(Zaridah et al. 2006). This same method will apply to the rest species of Ae. albopictus mosquito larvae.
Mortality (%) = (X-Y)/Y x 100 (2)
The formula above indicates X = percentage survival in the untreated control and Y = percentage
survival in the treated sample. Any small, unhealthy or damage larvae were removed or replaced. The
depth of the water in the cups or vessels was maintained between 5 cm to 10 cm because the deeper level
may cause undue mortality.
2.5. Data analysis
Data collected were pooled for analysis. LC50 and LC90 were calculated from a log dosage probit
mortality regression. The lethal concentration to 50% (LC50) and lethal 90% (LC90) of test organisms,
95% onfidence interval and their slopes of probit regression line will be determine using Microsoft excel
2010 (Zaridah et al. 2006).
3. Result
3.1. Phytochemical screening
Table 1. Result of phytochemical constituent
Plant species Phytochemical Petal Leaf Root
Ipomoea cairica Alkaloids + + -
Glycosides + - -
Saponins + + -
Phytosterols - - -
Phenols + - -
Tannins - + +
Flavanoids + + +
Protein and acid amino - + +
Diterpens - + +
Note: (+) Detected; (-) Not detected
The result of the phytochemical screening as tabulated in Table 1 reveals that flavonoids were present
in the petals, leaves and roots extract of Ipomoea cairica. Alkaloids were detected both in the petals and
leaves extract while glycosides were detected in the petals extract only. In addition, Saponins were
184 Ahmad Razali Ishak et al. / Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188
tected in both petals and leaves while phenols were detected only in the petals extract. For tannins, protein
and amino acids and diterpens were detected in both leaves and roots extract but not present in the petals.
Phytosterols test gave negative readings to all of the extracts.
3.2. Knockdown and cumulative mortality assessment
Figure 1 showed the comparison of cumulative knockdown rate between three different parts of
Ipomoea cairica extracts. Based on the trend, the petal’s extracts are more effective compared to the
leaves and roots extract to both species of Aedes mosquito. Figure 1 also showed the percentage mortality
of third instar larvae for Ae. aegypti and Ae. albopictus as larvacides. The results revealed that the
methanol extractions of Ipomoea cairica under laboratories conditions have indicated the potential
larvacidal activities. Furthermore, the treatment of increasing dosage of extracts (10, 25, 50, 75, 100 ppm)
clearly indicates the effect of the mortality dependence on the concentration of the plant extracts.
(b)
(a)
(c) (d)
Fig. 1. Trend of knockdown rate and cumulative mortality rate induced by crude extract of three different part of Ipomoea cairica on
the late third instar larvae. Noted (a&c) Ae.albopictus and (b&d) Ae.aegypti in different concentration exposure.
Ahmad Razali Ishak et al. / Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188 185
In addition, the comparison of cumulative larval mortalities between three different parts of Ipomoea
cairica extracts indicates that the leaves and petal extracts provide more effective result compared to root
extracts. The leaf extract was the most lethal although in a low concentration compared to other parts of
plants for both Ae. albopictus and Ae. aegypti larvae. However, at higher concentration, the petal extracts
were more lethal than leaves extracts.
3.3. Larvacidal efficacy
As shown in Table 2, all parts of Ipomoea cairica also exhibited strong larvacide activities against Ae.
aegypti and Ae.albopictus larvae in 48 hour exposure. The LC50 values of the leaf, petal and root essential
oils against Ae.albopictus were 20.5 mg/L (LC90: 62.5 mg/L), 27.9 mg/L (LC90:163.1mg/L) and 34.3
mg/L (LC90:105.3mg/L), respectively. On the other hand, the essential oil of leaves and petals was also
potent against Ae. aegypti larvae, showing LC50:12.7 mg/L (LC90:62.3 mg/L) and LC50:13.6 LC90:182.7
mg/L). The least larvacide activity was observed for the root extracts of Ipomoea cairica, with a LC50 =
31.9 mg/L (LC90:157.7 mg/L).
Table 2. Lethal concentration of larvacidal activity from different part of Ipomoea cairica against Ae aegypti and Ae. albopictus
Mosquito Treatment 24 hours 48 hours
species LC50 LC90 Regression LC50 LC90 Regression
(mg/L) (mg/L) (mg/L) (mg/L)
Petal 31.8 174.4 Y= 1.7317x + 2.3985 27.9 163.1 Y= 1.6687x + 2.5879
Aedes
Leaf 21.7 118.2 Y= 1.7378x + 2.6779 20.5 62.5 Y= 2.1195x + 2.2181
albopictus
Root 37.6 `114.8 Y= 2.6418x + 0.8379 34.3 105.3 Y= 2.6314x + 0.9586
Aedes Petal 18.0 180.4 Y= 1.2802x + 3.3916 13.6 182.7 Y= 1.1346x + 3.7138
aegypti Leaf 12.2 82.8 Y= 1.5363x + 3.3337 12.7 62.30 Y= 1.8573x + 2.9471
Root 37.5 121.1 Y= 2.5163x + 1.0381 31.9 157.7 Y= 1.8331x + 2.2513
LC 50: Lethal concentration of required to kill 50 per cent the population exposed
LC 90: Lethal concentration required to kill 90 per cent of the population exposed.
4. Discussion
The control of mosquito larvae using larvicidal agents is a major component in the control of vector
borne diseases. Thus, investigation into plants as potential larvicides is considered as viable and preferred
alternative in the control of the community level. Moreover, the plant should be locally available or easily
found at the local level. Phytochemicals compound act as general toxicants both against adult as well as
against larval stages of mosquitoes, while others interfere with growth and development or reproduction
or produce olfactory stimuli acting as repellent or attractant (Mathew et al., 2009). In this study, the
phytochemical screening test reveals the present of phytochemical constituents in all part of Ipomoea
cairica extracts. These phytochemicals found in this study was responsible as a larvacide agent to kill the
mosquito’s larvae (Kabaru & Gichia, 2009). The finding of the current study is consistent with Chapagain
et al. (2008) who discovered that the increasing concentration of root derived from callus saponins of B.
aegyptiaca increased the mortality of the larvae. Furthermore, it supported the idea of the saponin
compound in the plant can help to destroy the mosquito larvae. In addition, the finding of Mathivanan et
al. (2010) also suggested that the presence of alkaloid from Piper longum fruit was found to be active
against mosquito larvae of Cx. Pipiens. Many studies have been conducted on the knockdown effect of
adult mosquitoes (Xue et al. 2004), but few study had done of knockdown rate for larvae mosquitoes. In
this study, the knockdown effect occurred for all test concentration in all different parts of Ipomoea
cairica extraction. Knockdown is defined as the initial effect such as morbid or unusual behaviour due to
186 Ahmad Razali Ishak et al. / Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188
the alteration of a specific physiological process or processes that taken placed upon contact with the
toxicant (Hidayatulfathi et al, 2004). Before exposure, all the third instar larvae of both Aedes sp.
exhibited a natural behaviour with the siphon had pointed up through the water surface and heavy hung
down. However, after 5 minutes of exposure, abnormal evidence of excitation, restlessness, and
sluggishness was initially observed. Excitation and restlessness persisted for between 10-30 minutes, and
other abnormal motions were seen such as coiling movement. The treated larvae frequently sank down
and floated up again quickly. During the period of 30-60 minutes, some larvae showed more toxic
symptoms including vibration and convulsion at the bottom of the plastic cup. Afterwards, more larvae
exhibited the toxic symptoms. In the larvicidal bioassays, there were no pupation and mortalities that
exceeded 20% in the negative control solutions thus, discarding the bioassay experiments or correcting
the mortality data was not necessary. The result of LC50 and LC90 of all extracts, showed that the leaf of
Ipomoea cairica essential oil were more toxic to Ae.aegypti compared to Ae. albopictus. The least toxic to
both these species is the root extracts which probably contain slow-acting insecticidal ingredients. Jantan
et al. (2003) evaluated 17 methanol extracts and nine essential oils of Malaysian plants for their larvicidal
activity against Ae. aegypti. The essential oils of Cinnamomum impressicostatum, Cinnamomum
microphyllum and Curcuma domestica showed significant effects, reaching LC50 values of 13.7, 20.6 and
20.9 mg/ml, respectively. Except for the oil of Zingiber cassumunar, the essential oils of the other species
were effective against larvae, with LC50 values less than 200 mg/ml. Garcinia praniana, Garcinia
griffithii, Labisia pumila var. alata, L. pumila var. pumila and Mitragyna speciosa showed relatively high
activity with LC50 values ranging from 103–271 mg/ml. Although, Ipomoea cairica plant extracts have
been studied for various medicinal properties, there is no study conducted on the mosquito larvicidal
properties of this plant reported to our knowledge. From the results of the present investigation, the
larvicidal activities observed for all extraction (leaf, petal and root) are of special interest. The low LC 50
and LC90 indicating their high potential to serve as a larvicidal agent against key dengue vectors. Further
study on the isolation of the active principles responsible for the mosquito larvicidal activity may pave the
way for the development of a botanical insecticide for the control of Ae.aegypti and Ae.albopictus, the
dengue vector. At present, mosquito control is mostly directed against larvae because the fight against
adult is temporary, unsatisfactory and polluting the environment, but larvae treatment is more localized in
time and space resulting in less dangerous outcomes (Chowdhury et al., 2008). Larvae control can be an
effective control tool due to the low mobility of larvae mosquitoes, especially where the principal
breeding habitats are manmade and can be easily identified (Howard et al. 2007). These studies have
proven the principle in controlling larvae breeding using natural products is feasible and cost effective.
Eco-friendly chemicals are recommended in larviciding mosquito breeding sites. In this case, a plant
species that is used in managing of mosquitoes was analyzed for its larvicidal properties. Plants being a
natural source of compounds are known to contain larvicidal agents, which may act independently or in
combination, even in their semi purified form. The plant extract can be used together with chemical
larvicide to produce a synergistic effect which is more effective and reduce negative physiological impact
to the environment.
5. Conclusion
This study proof that the extracts of Ipomoea cairica has remarkable larvicidal properties and should
be further explore in the future to reduce the dependency on chemical larvicide such as Abate, Temephos
for mosquito control program. Further study is needed to extract Ipomoea cairica using different types of
solvents and identifying the active component of the plant that causes larvae mortality. Field trials are
also required to determine their potential as an alternative larvicide. Plant extracts could be an alternative
method for mosquito larvicide because they contain bioactive chemicals which are generally free from
Ahmad Razali Ishak et al. / Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188 187
harmful effects. This would reduce the use of chemicals which is hazardous to the living environment and
promote sustainable utilization for available bio-resources. Tropical country like Malaysia has advantage
because indigenous plants such as Ipomoea cairica are available in large quantities. Moreover, it is cost
effective to produce their extracts and for field use in vector control program.
Acknowledgement
The authors sincerely thank the Research Management Institute (RMI) for awarding research funding
from Universiti Teknologi MARA (UiTM) and Department of Higher Learning, Ministry of Education,
Malaysia.
References
Abu Bakar, A., Sulaiman, S., Omar, B., Mat Ali, R. 2012. Evaluation of Melaleuca cajuputi (Family; Myrtaceae) essential oil in
spray cans against dengue vectors in low cost housing flats. Journal Arthropod-Borne Dieases 6(1):28-35
Ananth, S., Thangamathi, P., Pazhanisamy, S., Meena, S. 2009. Larvicidal Activities of three medicinal Plants against the Filarial
Mosquito Culex Quinquefasciatus. Journal of Basic and Applied Biology 3(1 & 2):53-5
Azmathullah, N.M. 2011.Phytochemical screening of calotropis procera flower extracts and their Bio-control potential on Culex sp
mosquito larvae and pupae. International Journal of Pharmaceutical & Biologycal Archive 2(6):1718-1721
Borah,R., Kalita, M.C., Kar, A. & Talukdar, A.K. 2010. Larvicidal efficacy of Toddalia asiatica (Linn.) Lam against two mosquito
vector Aedes Aegypti and Culex quinquefasciatus. African Journal of Biotechnology 9(16):2527-2530.
Chapagain, Bishnu P, Saharan, Vinod, & Wiesman, Zeev. (2008). Larvicidal activity of saponins from Balanites aegyptiaca callus
against Aedes aegypti mosquito. Bioresource technology, 99(5), 1165-1168.
Choochote,W., Tuetub, B., Kanjanapothi, D., Rattanachanpichai, E., Chaithong, U., Chaiwong, P., Jitpakdi, A., Tippawangkosol, P.,
Rivong, D., Pitasawat, B. 2004. Potential of crude seed extracts of celery, Apium graveolens L., against the mosquito Aedes
aegypti (L.) (Diptera: Culicidae). Journal of Vector Ecology 29(2):340-6
Chowdhury, N., Ghosh, A., Chandra, G. 2006. Mosquito larvicidal activities of solanum villosum berry extraction against the
dengue vector stegomyia aegypti. BMC Complementary and Alternative Medicine 8:10
Gubler, D.J. & Clark, G.G. 1995. Dengue/dengue hemorrhagic fever: the emergence of a global health. Emerging Infectious Disease
1:55-57
Hidayatulfathi, O.,Sallehuddin, S., Shafariatul, A.I., Zaridah, M.Z. 2005. Studies on the Larvacidal , Adulticidal and repellent
activities of Acorus calamus Linn. Against Aedes Aegypti Linn. (Diptera: Culicidae) Anns. Medical Entomology 14(1):6-13.
Jantan, I., Wong, O.P., Sheila, D.V., Nazni, W.S., (2003), Larvicidal Activity of the Essential Oils and Methanol Extracts of
Malaysian Plants on Aedes aegypti, Pharmaceutical Biology, 2003, Vol. 41, No. 4 : Pages 234-236
Kabaru, JM, & Gichia, L. (2009). Insecticidal activity of extracts derived from different parts of the mangrove tree Rhizophora
mucronata (Rhizophoraceae) Lam. against three arthropods. African Journal of Science and Technology, 2(2).
Katade, S.R., Pawar, P.V., Wakharkar, R.D., Deshpande, N.R. 2006. Sterculia guttata seeds extractives – An effective mosquito
larvicide. Indian Journal of Experimental Biology 44:662-665.
Lam, S.K. 1993. Two decade of Dengue in Malaysia. Tropical Medicine, 35(4):195-200.
Mathew, Nisha, Anitha, MG, Bala, TSL, Sivakumar, SM, Narmadha, R, & Kalyanasundaram, M. (2009). Larvicidal activity of
Saraca indica, Nyctanthes arbor-tristis, and Clitoria ternatea extracts against three mosquito vector species. Parasitology
research, 104(5), 1017-1025.
Mathivanan, T, Govindarajan, M, Elumalai, K, Krishnappa, K, & Ananthan, A. (2010). Mosquito larvicidal and phytochemical
properties of Ervatamia coronaria Stapf.(Family: Apocynaceae). Journal Vector Borne Disease, 47, 178-180.
Misni, N., Sulaiman, S., Othman, H. 2008. The repellent Activity of Piper aduncum Linn (Family : Piperaceae) essential oil against
Aedes aegypti using human volunteers. Journal of Tropical Medicine & Parasitology 31:63-9
Misni, N., Sulaiman, S., Othman, H., Omar, B. 2009. Repellency of essential oil of Piper aduncum against Aedes albopictus in the
laboratory. Journal of American Mosquito Control Association 25 (4): 442:447
Nazri Che Dom., Ahmad, A. H., & Ismail, R. (2013a). Habitat Characterization of Aedes Sp. Breeding in Urban Hotspot
Area. Procedia-Social and Behavioral Sciences, 85, 100-109.
Nazri Che Dom., Ahmad, A. H., Ishak, A. R., & Ismail, R. (2013). Assessing the Risk of Dengue Fever Based On the
Epidemiological, Environmental and Entomological Variables. Procedia-Social and Behavioral Sciences, 105, 183-194.
188 Ahmad Razali Ishak et al. / Procedia - Social and Behavioral Sciences 153 (2014) 180 – 188
Prashant, T., Bimlesh K., Mandeep K., Gurpreet K., Harleen K., (2011) Phytochemical screening and Extraction: A Review, Journal
of Internationale Pharmaceutica Sciencia, Vol. 1 Issue 1
Raghavendra, B.S., Prathibha, K.P., & Vijayan, V.A. 2011. Larvicidal efficacy of Eugenia Jambolana Linn. Extracts in three
mosquito species at Rahman, W.A., Che’Rus, A. & Ahmad, A.H. (1997) Malaria and Anopheles mosquitoes in Malaysia
Southeast Asian .Journal of Tropical Medicine and Public Health 28:599-605.
Sulaiman, S., Abang Kamaruddin, D.S.F., Othman, H. 2008. Evaluation of Bifenthrin and Acorus calamus Linn. Extract against
Aedes aegypti L. and Aedes albopictus (Skuse). Iranian Journal Arthropod-Borne Disease 2(2): 7-11
Sulaiman, S., Othman, H., Shaari, N., Pawanchee, Z.A., Wahab, A., Abdullah, A.R.L. 2006. Field evaluation of Litsea elliptica
extract and bifenthrin by ULV spraying against dengue vectors. Journal of Tropical Medicine Parasitology 29:51-5
Xue, Rui-De, Ali, Arshad, & Barnard, Donald R. (2003). Laboratory evaluation of toxicity of 16 insect repellents in aerosol sprays
to adult mosquitoes. Journal-American Mosquito Control Association, 19(3), 271-274.
Zaridah, M.Z., Nor Azah M.A., Rohani A. 2006. Mosquitocidal activities of Malaysian plants. Journal of Tropical Forest Science,
18(1):74-80
