Entomological investigations into an epidemic of Japanese encephalitis (JE) in northern districts of West Bengal, India (2011-2012).

Background & objectives: Japanese encephalitis (JE) is one of the most important arboviral diseases of human beings with outbreaks in many parts of Southeast Asia including India. We present the entomological findings of an outbreak occurred in northern part of West Bengal during 2011-2012 with special emphasis on the role of JE vectors in different seasons. Methods: Adult mosquito collections were made with the help of mouth aspirators, aided by flash lights during day time resting inside human and animal habitations as indoor, and resting outside field grasses, bushes, underneath of culverts and bridges as outdoor, and in and around the pig enclosures and cattle sheds during dusk period in JE affected villages from Cooch Behar, Dakshin Dinajpur, Darjeeling and Jalpaiguri districts in North West Bengal. In all study villages, a long handled with enamel bowl dipper was used to obtain immature stages of mosquitoes from various breeding habitats. Results: A total of 19 different types of mosquito breeding habitats were examined for vectors of JE. From these habitats, 23.7 per cent were positive for breeding during the study period. Overall, nine different species were recorded through emergence, but none was positive for JE virus when subjected for detection of virus. Adult mosquitoes of more than 50 per cent of the potential JE vector species obtained through dusk and the rest through indoor and outdoor collections in all seasons. Altogether, 27 different species were recorded. Most of these were JE vectors. Interpretation & conclusions: Our results showed that in addition to Cx. vishnui subgroup, detection of JE virus antigen in Cx. quinquefasciatus indicated the possible maintenance of JE virus in nature through poor vector mosquitoes throughout the year. Since, all potential vector species reported elsewhere in India were also found in this region and fluctuated in density in different seasons, a proper integrated vector control programme needs to be implemented to control JE transmission.

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.

Quality problems of anti-tuberculosis Fixed-dose Combinations (FDCS) A way forward.

Fixed dose combinations (FDCs) of anti-tuberculosis drugs (Rifampicin, Isoniazid, Pyrazinamide and Ethambutol hydrochloride) have several advantages over loose combinations of these drugs. Hence, the use of FDCs in the short course chemotherapy of tuberculosis is being promoted internationally. However, these products are associated with quality problems such as loss of bio-availability of Rifampicin and instability of the drugs in the combination. Recent research in Indian laboratories has helped in identification of the major reasons behind these problems, which suggest that an entirely different approach is required to that currently being recommended by the world bodies. The purpose of this article is to briefly discuss the twin quality problems and to highlight the potential forward ways.

Utility of expanded polystyrene (EPS) beads in the control of vector-borne diseases.

The use of chemicals or bio-larvicides for the control of Culex quinquefasciatus and Anopheles stephensi breeding in pit latrines and overhead tanks (OHT) respectively is discouraged owing to many undesirable impacts in the environment. Due to faecal contamination and poor survival, use of predatory fish in OHTs is not feasible. The use of expanded polystyrene (EPS) beads is a potential alternative in these habitats. EPS beads not only prevent oviposition but also kill the immature by forming a thick blanket on the water surface. A thick layer of 2 cm with beads of 2 mm is sufficient to suppress and prevent mosquito breeding. These are cheap, environmentally safe and do not need frequent application since they remain on the surface for quiet a long time. Successful trials against C. quinquefasciatus breeding in pit latrines, soakage pits, septic tanks, etc., have been carried out in Kenya, Zimbabwe and Tanzania. Certain trials with EPS indicated reduction in microfilaria (mf) rate besides decline in biting density. In India, EPS beads have also been used on small scale for the control of A. stephensi and A. culicifacies breeding in OHTs and unused wells respectively. The polystyrene beads have also been reported to be effective in the control of mosquito breeding in biogas plants and other industrial situations. The practical utility of EPS beads in the control of vector-borne diseases has been discussed in the present review.

Field evaluation of Spicbiomoss, a biolarvicidal formulation of Bacillus sphaericus against immatures of Culex quinquefasciatus & Anopheles species.

Spicbiomoss, an aqueous suspension formulation of Bacillus sphaericus was evaluated for its efficacy against immatures of Culex quinquefasciatus at the application rate of 120 l/ha in cement tanks, cesspits and drains in Pondicherry and in drains in Kochi, Kerala. The formulation was also tested against anophelines (Anopheles fluviatilis and An. culicifacies) breeding in stream pools in Malkangiri district, Orissa. In cement tanks and cesspits more than 80 per cent reduction in immature density was observed for a period of 6-13 days (mean 9.8 +/- 1.25 days) and 3-8 days (mean 5.2 +/- 0.7 days) respectively. The same level of reduction was found to last for 1-4 days (mean 2.2 +/- 0.52 days) in drains in Pondicherry and 2-9 days (mean 4.8 +/- 1.17 days) in Kochi. In bunded stream pools 40-79 per cent reduction in immature density of Anopheles was obtained for an average period of 1-7 (mean 3.83 +/- 0.98) days. There was no improvement in the efficacy of the formulation against anophelines even at the higher application rate (240 l/ha). Thus, Spicbiomoss can be used against Cx. quinquefasciatus in an integrated vector management programme.

Mattancherry urban agglomeration, a diminishing focus of lymphatic filariasis in Kerala.

Parasitological survey conducted recently in Mattancherry, Kerala showed a low prevalence of microfilariae (mf), with a mf rate of 1.81%, compared to that of 1956 when the mf rate was 14.7%. No brugian infection was encountered in the present investigation, though both bancroftian and brugian infections were reported earlier. Culex quinquefasciatus, the vector of bancroftian filariasis was the most predominant (93.3%) mosquito species, and its infection and infectivity rates were 0.76% and 0.54% respectively. Mansonioides spp., the vectors of brugian filariasis constituted only 0.2% of the total mosquitoes and with no infection. The overall reduction in the prevalence of mf cases and vector infection rates over the years is attributed to the continued activities of the National Filaria Control Programme. The disappearance of brugian filariasis could mainly be due to the drastic reduction in the Mansonioides breeding habitats, as a result of rapid urbanization coupled with improved socio economic conditions.

Brackish water mosquito problem of Vypeen Island, Cochin, Kerala.

A preliminary study has shown that the marshy terrain and brackish water bodies associated with mangrove forests contributed profuse breeding of mosquitos in Vypeen island, causing a severe menace to the island population. A total of 14 species belonging to four genera viz, Aedes, Anopheles, Armigeres and Culex was recorded from different habitats. Culex sitiens was found to be the predominant mosquito in all the perennial breeding habitats. The extent of different habitats in the production of mosquitos, and its possible abatement, using environmental and/or biocontrol methods are discussed.

Prevalence of bancroftian filariasis & its control by single course of diethyl carbamazine in a rural area in Tamil Nadu.

The prevalence of microfilaraemia, clinical spectrum of bancroftian filariasis and vector potential were studied in Vettavalam village in North Arcot district of Tamil Nadu. The effectiveness of selective therapy with diethyl carbamazine (DEC) in controlling filariasis in rural areas was also evaluated. The prevalence of microfilaraemia (mf rate) and disease (disease rate) was found to be 11.7 and 11.09 per cent respectively. Hydrocele was the dominant clinical sign in males and lymphoedema in females. The density of the vector Culex quinquefasciatus was 25.44 females/man-hour, and the infection and infectivity rates were 18.16 and 1.09 per cent respectively. All mf carriers detected after the mass blood survey were given a single course of DEC at the dosage of 6 mg/kg of body wt/day for 12 days. Only 61.6 per cent of them took the full course of DEC treatment. DEC therapy brought down the mf rate from 11.7 to 5.84 per cent after one month. In the absence of further treatment, there was no significant change in mf prevalence after one year.