Screening for Larvicidal Activity of Aqueous and Ethanolic Extracts of Fourteen Selected Plants and Formulation of a Larvicide against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) Larvae

This study aims to: a) obtain ethanolic (95% EtOH) and aqueous extracts of Selaginella elmeri, Christella dentata, Elatostema sinnatum, Curculigo capitulata, Euphorbia hirta, Murraya koenigii, Alpinia speciosa, Cymbopogon citratus, Eucalyptus globulus, Jatropha curcas, Psidium guajava, Gliricidia sepium, Ixora coccinea and Capsicum frutescens and screen them for larvicidal activities against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) larvae; b) to fractionate the most active extract and determine the most active fraction; c) to determine the larvicidal properties of the most active extract and fraction against by computing their percentage mortality, LC50, and LC90 after 24 and 48 hours of exposure; and d) to determine the nature of the components of the active extracts and fractions using phytochemical screening. Ethanolic (95% EtOH) and aqueous extracts of the selected plants will be screened for potential larvicidal activity against Ae. aegypti and Ae. albopictus using standard procedures and 1% malathion and a Piper nigrum based ovicide-larvicide by the Department of Science and Technology as positive controls. The results were analyzed using One-Way ANOVA with Tukey’s and Dunnett’s test. The most active extract will be subjected to partial fractionation using normal-phase column chromatography, and the fractions subsequently screened to determine the most active fraction. The most active extract and fraction were subjected to dose-response assay and probit analysis to determine the LC50 and LC90 after 24 and 48 hours of exposure. The active extracts and fractions will be screened for phytochemical content. The ethanolic extracts of C. citratus, E. hirta, I. coccinea, G. sepium, M. koenigii, E globulus, J. curcas and C. frutescens exhibited significant larvicidal activity, with C. frutescens being the most active. After fractionation, the ethyl acetate fraction was found to be the most active. Phytochemical screening of the extracts revealed the presence of alkaloids, tannins, indoles and steroids. A formulation using talcum powder–300 mg fraction per 1 g talcum powder–was made and again tested for larvicidal activity. At 2 g/L, the formulation proved effective in killing all of the test larvae after 24 hours.

Chikungunya Protease Domain–High Throughput Virtual Screening

Chikungunya virus (CHICKV) is an arboviruses belonging to family Tagoviridae and is transmitted to human through by mosquito (Aedes aegypti and Aedes albopictus) bite. A large outbreak of chikungunya has been reported in India between 2006 and 2007, along with several other countries from South-East Asia and for the first time in Europe. It was for the first time that the CHICKV outbreak has been reported with mortality from Reunion Island and increased mortality from Asian countries. CHICKV affects all age groups, and currently there are no specific drugs or vaccine to cure the disease. The need of antiviral agents for the treatment of CHICKV infection and the success of virtual screening against many therapeutically valuable targets led us to carry out the structure based drug design against Chikungunya nSP2 protease (PDB: 3TRK). Highthroughput virtual screening of publicly available databases, ZINC12 and BindingDB, has been carried out using the Openeye tools and Schrodinger LLC software packages. Openeye Filter program has been used to filter the database and the filtered outputs were docked using HTVS protocol implemented in GLIDE package of Schrodinger LLC. The top HITS were further used for enriching the similar molecules from the database through vROCS; a shape based screening protocol implemented in Openeye. The approach adopted has provided different scaffolds as HITS against CHICKV protease. Three scaffolds: Indole, Pyrazole and Sulphone derivatives were selected based on the docking score and synthetic feasibility. Derivatives of Pyrazole were synthesized and submitted for antiviral screening against CHICKV.

Mosquitoes Larval Breeding Habitat in Urban and Suburban Areas, Peninsular Malaysia

Larval survey was carried out in 6 localities in the urban areas (Putrajaya) and suburban areas (Kuala Selangor) from January until December 2010. A total of 520 representative households in 6 localities were selected. Breeding habitats were sampled outdoors in the surroundings of housing areas. The study indicated that the most predominant species found in both areas was Aedes albopictus with the gardening utensil as a preferred breeding microhabitat for Putrajaya, in contrast to the artificial containers for Kuala Selangor. From a total of 1083 mosquito larvae species, 984 were Aedes albopictus larvae, 67 positive larvae of Aedes aegypti and 32 of Culex larvae. Aedes Index and Container Index were elevated in Putrajaya with 13% and 11% respectively which is higher than the standard given by the Ministry of Health, Malaysia. This results implicating dengue-sensitive skewed to the urban areas. Breteau Index result also above the standard in both study locations.

Seasonal Prevalence of Aedes aegypti and Ae.albopictus in Three Topographical Areas of Southern Thailand

This study investigated the seasonal prevalence of Aedes aegypti and Ae. albopictus larvae in three topographical areas (i.e. mangrove, rice paddy and mountainous areas). Samples were collected from 300 households in both wet and dry seasons in nine districts in Nakhon Si Thammarat province. Ae. aegypti and Ae. albopictus were found in 21 out of 29 types of water containers in mangrove, rice paddy and mountainous areas. Ae. aegypti and Ae. albopictus laid eggs in different container types depending on season and topographical areas. Ae. aegypti larvae were found most in metal box in mangrove and mountainous areas in wet season. Ae. albopictus larvae were also found most in metal box in mangrove and mountainous areas in both wet and dry seasons. All Ae. albopictus larval indices were higher than Ae. aegypti larval indices in all three topographical areas and both seasons. HI and BI did not differ in three topographical areas but differed between Aedes sp. HI for both Ae. aegypti and Ae. albopictus in all three topographical areas in both seasons were greater than 10 %, except Aedes aegypti in rice paddy area in wet season. This indicated high risks of DHF transmission in these areas.

Larval Occurrence and Climatic Factors Affecting DHF Incidence in Samui Islands, Thailand

This study investigated the number of Aedes larvae, the key breeding sites of Aedes sp., and the relationship between climatic factors and the incidence of DHF in Samui Islands. We conducted our questionnaire and larval surveys from randomly selected 105 households in Samui Islands in July-September 2006. Pearson-s correlation coefficient was used to explore the primary association between the DHF incidence and all climatic factors. Multiple stepwise regression technique was then used to fit the statistical model. The results showed that the positive indoor containers were small jars, cement tanks, and plastic tanks. The positive outdoor containers were small jars, cement tanks, plastic tanks, used cans, tires, plastic bottles, discarded objects, pot saucers, plant pots, and areca husks. All Ae. albopictus larval indices (i.e., CI, HI, and BI) were higher than Ae. aegypti larval indices in this area. These larval indices were higher than WHO standard. This indicated a high risk of DHF transmission at Samui Islands. The multiple stepwise regression model was y = –288.80 + 11.024xmean temp. The mean temperature was positively associated with the DHF incidence in this area.