Seasonal abundance & role of predominant Japanese encephalitis vectors Culex tritaeniorhynchus & Cx. gelidus Theobald in Cuddalore district, Tamil Nadu.

Background & objectives: Japanese encephalitis (JE) is the leading cause of viral encephalitis in Asia. The first major JE outbreak occurred in 1978 and since 1981 several outbreaks had been reported in the Cuddalore district (erstwhile South Arcot), Tamil Nadu, India. Entomological monitoring was carried out during January 2010 - March 2013, to determine the seasonal abundance and transmission dynamics of the vectors of JE virus, with emphasis on the role of Culex tritaeniorhynchus and Cx. gelidus. Methods: Mosquito collections were carried out fortnightly during dusk hours in three villages viz. Soundara Solapuram, Pennadam, Erappavur of Cuddalore district. Mosquitoes were collected during dusk for a period of one hour in and around the cattle sheds using oral aspirator and torch light. The collected mosquitoes were later identified and pooled to detect JE virus (JEV) infection by enzyme linked immunosorbent assay (ELISA). Results: A total of 46,343 mosquitoes comprising of 25 species and six genera were collected. Species composition included viz, Cx. tritaeniorhynchus (46.26%), Cx. gelidus (43.12%) and other species (10.62%). A total of 17,678 specimens (403 pools) of Cx. gelidus and 14,358 specimens (309 pools) of Cx. tritaeniorhynchus were tested, of which 12 pools of Cx. gelidus and 14 pools of Cx. tritaeniorhynchus were positive for JE virus antigen. The climatic factors were negatively correlated with minimum infection rate (MIR) for both the species, except mean temperature (P<0.05) for Cx. gelidus. Interpretation & conclusions: High abundance of Cx. tritaeniorhynchus and Cx. gelidus was observed compared to other mosquito species in the study area. Detection of JEV antigen in the two species confirmed the maintenance of virus. Appropriate vector control measures need to be taken to reduce the vector abundance.

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.

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.

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).

Studies on community knowledge and behavior following a dengue epidemic in Chennai city, Tamil Nadu, India

In 2001, a major dengue outbreak was recorded in Chennai city, with 737 cases (90%) out of a total of 861 cases recorded from Tamil Nadu state. A KAP survey was carried out to assess the community knowledge, attitude and practice on dengue fever (DF), following the major dengue outbreak in 2001. A pre- tested, structured questionnaire was used for data collection. The multistage cluster sampling method was employed and 640 households (HHs) were surveyed. Among the total HHs surveyed, 34.5% of HHs were aware of dengue and only 3.3% of HHs knew that virus is the causative agent for DF. Majority of the HHs (86.5%) practiced water storage and only 3% of them stored water more than 5 days. No control measures were followed to avoid mosquito breeding in the water holding containers by majority of HHs (65%). Sixty percent of HHs did not know the biting behaviour of dengue vector mosquitoes. The survey results indicate that the community knowledge was very poor on dengue, its transmission, vector breeding sources, biting behavior and preventive measures. The lack of basic knowledge of the community on dengue epidemiology and vector bionomics would be also a major cause of increasing trend of dengue in this highly populated urban environment. There is an inevitable need to organize health education programmes about dengue disease to increase community knowledge and also to sensitize the community to participate in integrated vector control programme to resolve the dengue problem.

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).

Natural vertical transmission of dengue viruses by Aedes aegypti in Chennai, Tamil Nadu, India.

BACKGROUND & OBJECTIVE: Dengue viruses are spread and maintained in an Aedes aegypti-human- Ae. aegypti cycle in urban areas of the tropics. Dengue viruses are also maintained in nature by vertical transmission by Ae. aegypti. A study was undertaken in Chennai, a known endemic city in south India, to comprehend the natural vertical transmission dynamics in Ae. aegypti and to assess its epidemiological importance. METHODS: Ae. aegypti males collected in resting and landing collections were tested for dengue virus infection by antigen-capture enzyme-linked immunosorbent assay (ELISA) and further examined by insect bioassay, Toxorhynchites splendens inoculation-indirect immunofluorescence technique (Toxo-IFA) using serotype-specific monoclonal antibodies (Mabs), if found positive by ELISA. RESULTS: Of the 509 pools of Ae. aegypti males (n=5408) screened, 15 pools, collected in April, June- July, November-December in 2003 and March, May in 2004, were found positive for dengue virus infection and the minimum infection rate (MIR) among adult males was high in June 2003 (28.0/ 1000). Three positive pools could be serotyped as dengue-2 (2 pools) and dengue-3 (1 pool). INTERPRETATION & CONCLUSION: Dengue virus isolations from wild caught males of Ae. aegypti indicate the occurrence of transovarial transmission. Vertical transmission was mainly observed in summer months when dengue infections in humans were low suggesting that dengue viruses adopt a novel strategy of surviving adverse climatic conditions.

Diagnostic methods for detection & isolation of dengue viruses from vector mosquitoes.

Dengue is a deadly mosquito-borne infection warranting urgent attention for its containment particularly in the tropical and subtropical countries. In the absence of a vaccine or any specific drug for its treatment, an early diagnosis is considered indispensable to prevent any casualty. Detection of viruses in human sera particularly in endemic areas is cumbersome, difficult and also not desirable. Therefore, as an alternative approach, detection of the dengue virus antigen in mosquitoes has provided a reliable tool to (i) comprehend the types of viruses circulating in nature; and (ii) help in designing vector-specific control strategies. A mélange of diagnostic techniques are currently available with some advantages or disadvantages. Traditionally, cell cultures and suckling mice have been employed for virus isolations. While the virus isolation method in baby mice is time consuming, slow and expensive, the mosquito cell cultures offer a good degree of specificity. Mosquito inoculation techniques have been reported for detection and propagation of flaviviruses. Though this technique is sensitive for routine virological confirmation of dengue fever, it requires large number of infected mosquitoes, besides being time consuming. Insect bioassays (Toxo-IFA) are generally cumbersome requiring special facilities and are not suitable for large-scale epidemiological surveillance. ELISA has been shown to be a rapid and sensitive alternative to insect bioassays for monitoring arboviruses in wild populations. Reverse transcriptase polymerase chain reaction (RT-PCR) is a recent molecular diagnostic technology used for detecting virus infections in mosquitoes, which gives rapid results but is expensive and prone to contamination. This review describes the development of various techniques involved in detection and isolation of dengue viruses in mosquitoes. Definite diagnosis of the impending dengue epidemic can be made using ELISA for virological surveillance system on dengue virus antigen in the mosquito vectors. Therefore, ELISA offers a potential tool and a convenient system for quickly screening large number of samples up to the serotype level which can be employed effectively and efficiently for large scale dengue surveillance programmes on wild caught mosquito vectors. ELISA positive samples can be screened further by Toxo-IFA system for virus isolation. On the other hand, techniques like mosquitoes cell culture, mosquito inoculation (Toxo-IFA) and RT-PCR techniques can be employed for dengue virus amplification.

Malaria outbreak in the Indira Gandhi Nahar Pariyojna command area in Jaisalmer district, Thar Desert, India.

A focal outbreak of malaria occurred in the villages situated close to the main Indira Gandhi canal near Ramgarh in Jaisalmer district, western Rajasthan. Stagnation of water over a month's period in the main canal as well as long standing rain water in the form of expansive lakes near these villages formed vast breeding grounds for the vectors like Anopheles culicifacies, along with A. stephensi already breeding in the 'tanka' and 'beri' in the epidemichit villages. Rapid mass blood surveys along with other entomological and parasitological investigations were conducted in four of the ten affected villages, viz., Seuva, Raghwa, Raimala and Sadhna. A total of 992 specimens belonging to four vector species were sampled, namely, A. stephensi (47.4%), A. culicifacies (41.0%), A. subpictus (11.2%) and A. annularis (0.4%). Epidemiologically, about one-fourth of the examined persons were positive (SPR 25.5%), although Plasmodium falciparum dominated the parasitaemia (49.5%). Available data are indicative of changed malariological scenario in the Indira Gandhi Nahar Pariyojna command area, where epidemics are regular features every year.

A study of an epidemic of acute respiratory disease in Jaipur town.

AIM: To detect an association between the sudden epidemic with respiratory symptoms, and fogging with dichlorovos in Jaipur town and to find out probable mechanism of causation of the epidemic. SUBJECTS AND METHODS: In this community based study of the epidemic, house to house survey of households selected using systematic random sampling was carried out. The incidence in the exposed and unexposed population, the relative risk and attributable risk were calculated. RESULTS: The incidence of cases was high (58.9%) in subjects present on roads at the time of fogging as compared to in those who were inside rooms of the houses (5.4%) and in those who were not in the locality at that time (1.8%) [Relative Risk (RR)=32.7 and Attributable Risk (AR)=96.9%]. CONCLUSION: High RR and AR in the present epidemic indicate strong association between fogging and occurrence of symptoms. In absence of signs and symptoms of organophosphorus poisoning it suggests that this could have been due to an inappropriate solvent or defective functioning of fog generator, leading to generation of an unusual dark fog, that might have irritated eyes and respiratory tract of exposed residents.

Knowledge, attitude and practice towards malaria in rural communities of the epidemic-prone Thar Desert, northwestern India.

In order to assess the knowledge, attitude and practice of the rural Thar Desert population about malaria and its vectors, a total of 345 heads of households, including housewives, suffering with malaria were surveyed randomly, along with the same number of healthy control subjects, in four villages of Jodhpur and Jaisalmer districts of Rajasthan, north-western India. Results showed that in the traditionally hypoendemic Thar Desert, with low levels of immunity against malaria particularly Plasmodium falciparum, a large segment of rural people were equally less aware about the dangers of the disease. About three-fourths of the studied population did not quite comprehend the purpose of the five decade-old National Anti Malaria Programme, while nearly 50% respondents were reluctant to accept the programme lacking the component of community participation but directly targetting on the control of the disease. Consequently, about 50% malaria patients and 60% healthy subjects lacked in taking initiative to resort to any anti-mosquito or anti-malaria measure to protect themselves, although 27% malaria patients and 2.9% healthy subjects acknowledge correct causes of malaria. This study, being the first of its kind in a desert community currently facing emergence of P. falciparum-dominated malaria outbreaks, is a testimony to the hitherto prevalent deep gaps in knowledge about the disease, and its results warrant sincere efforts to link up the malaria control programme with the community participation for protection against this scourage in future.

Effect of permethrin impregnated bednets in reducing population of malaria vector Anopheles culicifacies in a tribal village of Orissa state (India).

The role of permethrin impregnated bednets at the application rate of 0.5 g (ai)/m2 in reducing A. culicifacies population in a tribal village of Orissa, highly endemic for Plasmodium falciparum was studied. The per man-hour density was significantly reduced in the Kenduguda village for about three months where impregnated bednets were supplied to the villagers, while in the control village Deulaguda where unimpregnated nets were supplied, vector density remained high during the study period. Bioassay carried out on the treated nets showed that there was 100 per cent mortality in A. culicifacies for over three months with the unwashed nets as well as with bednets washed once or twice.