Characterization of the anopheline vector breeding habitats: Implications for elimination of malaria in tribal inhabited Car Nicobar Island

Background & objectives: Malaria is an important public health problem in Andaman & Nicobar archipelago. Among the three districts, Nicobar is the most endemic district where API is >2. In this district, the malaria incidence in Car Nicobar Tehsil has declined steadily over the past 10 years. A renewed initiative to consolidate this gain is being made with the ultimate objective of achieving zero indigenous transmission of malaria in Car Nicobar. So, the present study undertook a close environmental monitoring of water bodies for assessing changes in the risk potential of mosquito vector breeding habitats which can augment the elimination programme. Methods: The breeding habitats of anopheline mosquitoes were sampled in 16 areas of Car Nicobar Island for eight time periods during 2017-2020. Along with anophelines, various associated water parameters (n=60) were estimated, viz. physicochemical (n=13), and biological, which included culicine mosquito immatures, insect predators (n=5), phytoplanktons (n=31) and zooplanktons (n=10). Results: In the 16 study sites, overall 1126 surface water stagnating bodies constituting 21 different habitat types were surveyed. Of these, 17 were positive for anopheline breeding. Water bodies from three villages were consistently found to be positive for anopheline breeding. However, early instars of anopheline larvae were more abundant compared to the late instars. Four anopheline species were recorded, including Anopheles sundaicus, A. barbirostris, A. insulaeflorum and A. subpictus, in which 48 per cent were A. sundaicus. Multivariable analysis indicated that anopheline density was significantly higher in permanent water bodies than in temporary habitats (P<0.05) (high risk of anophelines). The highest pH (?8.2), dissolved solids (?0.39) levels showed significantly (P<0.05) decreased larval densities (lower risk of breeding), adjusted with breeding sites and season. Nitrite levels increased (P=0.022) larval densities. Interpretation & conclusions: The present study facilitated estimating the productive period of a larval habitat enabling target larval sources to reduce adult populations. Implementing larviciding strategy before monsoon season is presumably the most cost-effective strategy. The output can be utilized for environmental monitoring of mosquito breeding risk in other malaria endemic areas, particularly where medium/large water bodies are the predominant breeding sites for malaria vectors.

Impact on prevalence of intestinal helminth infection in school children administered with seven annual rounds of diethyl carbamazine (DEC) with albendazole.

Background & objectives: One third of the world’s population is infected with one or more of the most common soil-transmitted helminths (STH). Albendazole (ALB) is being administered with diethyl carbamazine (DEC) in filariasis endemic areas to eliminate lymphatic filariasis (LF) and helminth infections. In this study, the cumulative impact of seven annual rounds of mass drug administrations (MDA) of DEC and ALB on STH infection in school children in selected villages in southern India was determined. Methods: During 2001-2010, seven MDAs were implemented by the Tamil Nadu state health department, India. LF and STH infections were monitored in school children from 18 villages of the two treatment arms (viz, DEC alone and DEC+ALB). Kato-Katz cellophane quantitative thick smear technique was employed to estimate STH infections at three weeks, six months and one year post MDA. Results: Prior to treatment, an overall STH prevalence was 60 per cent. After each MDA, infection markedly reduced at three weeks post-treatment in both the arms. The prevalence increased at six months period, which was maintained up to one year. After seven rounds of MDA, the infection reduced from 60.44 to 12.48 per cent in DEC+ALB arm; while the reduction was negligible in DEC alone arm (58.77 to 52.70%). Interpretation & conclusions: Seven rounds of MDA with DEC+ALB reduced the infection load significantly, and further sustained low level of infection for 10 years. However, complete parasite elimination could not be achieved. To curtail STH infection in the community, MDA should be regularized and environmental sanitation measures need to be improved by effective community-based campaigns.

Detection of Wolbachia endobacteria in Culex quinquefasciatus by Gimenez staining and confirmation by PCR.

Background & objectives: Wolbachia are common intracellular bacteria that are found in arthropods and nematodes. These endosymbionts are transmitted vertically through host eggs and alter host biology in diverse ways, including the induction of reproductive manipulations, such as feminization, parthenogenesis, male killing and sperm-egg incompatibility. Since they can also move horizontally across species boundaries, Wolbachia is gaining importance in recent days as it could be used as a biological control agent to control vector mosquitoes or for paratransgenic approaches. However, the study of Wolbachia requires sophisticated techniques such as PCR and cell culture facilities which cannot be affordable for many laboratories where the diseases transmitted by arthropod vectors are common. Hence, it would be beneficial to develop a simple method to detect the presence of Wolbachia in arthropods. Method: In this study, we described a method of staining Wolbachia endobacteria, present in the reproductive tissues of mosquitoes. The reliability of this method was compared with Gram staining and PCR based detection. Results: The microscopic observation of the Gimenez stained smear prepared from the teased ovary of wild caught and Wolbachia (+) Cx. quinquefasciatus revealed the presence of pink coloured pleomorphic cells of Wolbachia ranging from cocci, comma shaped cells to bacillus and chain forms. The ovaries of Wolbachia (–) cured mosquito did not show any cell. Although Gram’s staining is a reliable differential staining for the other bacteria, the bacterial cells in the smears from the ovaries of wild caught mosquitoes did not take the stain properly and the cells were not clearly visible. The PCR amplified product from the pooled remains of wild caught and Wolbachia (+) Cx. quinquefasciatus showed clear banding, whereas, no banding was observed for the negative control (distilled water) and Wolbachia (–) Cx. quinquefasciatus. Interpretation & conclusion: The Gimenez staining technique applied, could be used to detect the members of the endobacteria Wolbachia easily, even in a simple laboratory without any special facilities or even in the field condition and for handling large number of samples in a shorter duration.

Wolbachia endobacteria in a natural population of Culex quinquefasciatus from filariasis endemic villages of south India and its phylogenetic implication

Understanding Wolbachia mosquito interactions have been recognized as an important concept to develop novel vector control strategies. The prevalence of Wolbachia endobacteria in a natural population of the filariasis vector Culex quinquefasciatus was determined by the polymerase chain reaction method. Earlier workers had estimated the infection rates of Wolbachia with only one or very few individuals per species. In our study large number of specimens were assayed, and a total of 750 adult Culex quinquefasciatus mosquitoes were collected from three south Indian villages of Tirukoilur and Mugaiyur blocks, monthly for a period of five months (December 2006 to April 2007) and screened for the presence of Wolbachia. The percentage prevalence in adult males ranged from 88% to 96%; while in females from 84% to 100%. An overall prevalence of 91.2% was observed. There was no significant difference observed in the proportion of mosquitoes positive for Wolbachia between males and females, and also between different months of the survey; except during the month of February ‘07. The wsp gene sequence of the Wolbachia strain of Cx. quinquefasciatus detected was BLAST analysed and showed 99% sequence similarity with Wolbachia sp. of Culex pipiens isolated from different geographical regions. Phylogenetic analysis based on wsp gene fragments showed that the present Wolbachia isolate was closely related with Wolbachia from Culex pipens pipiens, Niphotettix virescens (Order: Hemiptera) and Cnaphalocrosis medinalis (Order: Lepidoptera).

A study on chikungunya outbreak during 2007 in Kerala, south India.

Background & objectives: The first chikungunya outbreak occurred in Kerala during 2006 affecting 14 districts, followed by another during May 2007 affecting almost whole of the State. Four of the worst affected districts viz, Pathanamthitta, Idukki, Kottayam and Thrissur were surveyed during 2007 to understand the magnitude of the problem of chikungunya fever, particularly clinical signs and symptoms. Methods: A total of 1265 persons from 310 houses were surveyed door-to-door in 20 different localities representing four affected districts. The history and examination findings from 354 clinically diagnosed chikungunya cases were recorded. The symptoms recorded were fever, headache, myalgia, arthralgia, itch/rash, oedema, eye congestion, eye pain, oral ulcers, distaste, nausea, vomiting and haemorrhage. Results: The major symptoms were fever (100%), headache (97.5%), arthralgia (99.4%) and myalgia (99.4%). A significant difference was observed in oedema, distaste, nausea and headache among different age groups and these symptoms were reported to be lower (12.2-89.8%) in younger age group than in older age group (90.4-100%). No genderwise difference was observed for any of the symptoms. In clinically diagnosed chikungunya cases higher age group (>35 years) found with higher rate of severity with symptoms of oedema, distaste, nausea and headache when compared with lower age group (1-35 yr). Interpretation & conclusions: Chikungunya invaded Kerala State for the first time in 2006 and continues to be a major vector borne disease in the State. The clinical symptoms in affected cases highlighted high fever, sever myalgia and prolonged arthralgia, with occasional history of skin itch/rash (petechiae).

The role of urea in the oviposition behaviour of japanese encephalitis vectors in rice fields of South India

The effect of urea on the oviposition behaviour of culicine vectors of Japanese encephalitis was studied in rice fields. Gravid females had a strong preference for oviposition in urea treated areas in rice fields, while no such preference was exhibited in untreated areas. The egg laying declined in the area where urea treated water surface had a mechanical barrier, which allowed volatile fractions to escape, but prevented contact with the water. Urea was shown to act as an oviposition attractant/stimulant for Culex tritaeniorhynchus, but its role was not clear for Cx. vishnui, as the number of egg rafts obtained for the latter species was low