Investigation of a Chikungunya-like illness in Tirunelveli district, Tamil Nadu, India 2009-2010.

OBJECTIVE: To identify the aetiological agent/s of an outbreak of chikungunya-like illness with high morbidity and several fatalities in Tamil Nadu, India, 2009-2010. METHODS: Two hundred and seventeen serum samples were collected from the affected areas and screened for chikungunya virus (CHIKV), dengue virus (DENV) and Japanese encephalitis virus (JEV) IgM antibodies using MAC-ELISA kits. A few selected samples were also tested for Ross River, Sindbis, and Murrey Valley viruses by RT-PCR and Hantan virus by serology. Twelve acute serum and mosquito samples were processed for virus isolation in C6/36 cells. CHIKV isolate was characterised by RT-PCR and sequencing. RESULTS: Diagnostic levels of IgM antibodies were detected in 107 (49.3%) CHIKV samples and 22 (10.1%) DENV samples. IgM antibodies against JEV were not detected (n=46). Characterisation of the CHIKV isolate at genetic level demonstrated it as ECSA (E1: 226A). Thirty-six selected samples were also negative for Ross River, Sindbis, Murrey Valley and Hantan viruses. CONCLUSION: High prevalence of CHIKV IgM antibody positivity, clinical symptoms, virus isolation and the presence of vector mosquitoes clearly suggest CHIKV as the aetiological agent responsible for the outbreak.

Japanese encephalitis virus infection in mosquitoes reared from field-collected immatures and in wild-caught males.

Immature stages of mosquitoes collected in JE endemic areas of Karnataka, India between 1985 and 1987 were reared to adults and processed for the detection and isolation of Japanese encephalitis (JE) virus in an attempt to find naturally occurring vertical transmission of the virus. Males collected during 1985-1986 were also processed. A total of 15,785 adults reared from immatures and divided into 445 pools and 1,756 wild-collected males divided into 128 pools were processed using mosquito inoculation and immunofluorescence techniques. JE virus antigen was detected in 9 pools, 4 of which yielded JE virus. These were 2 pools of males and 1 pool of female Culex tritaeniorhynchus and 1 pool of male C. pseudovishnui, suggesting vertical transmission of JE virus in the mosquitoes.

Isolation of Japanese encephalitis virus from mosquitoes collected in Karnataka state, India from 1985 to 1987.

Detection and isolation of Japanese encephalitis (JE) virus using mosquito inoculation and immunofluorescence techniques were attempted from female mosquitoes collected in JE endemic areas of Kolar and Mandya districts of Karnataka state, India, from 1985 to 1987. 65,388 mosquitoes consisting of 19 species in 1541 pools were processed. Of these, 18 pools showed the presence of JE virus antigen. JE virus was isolated from 9 pools, 3 of Culex gelidus, 2 of C. tritaeniorhynchus, and one each of C. quinquefasciatus, C. fuscocephala, C. vishnui and Anopheles peditaeniatus. Isolation of JE virus from C. gelidus, C. fuscocephala, C. quinquefasciatus and An. peditaeniatus is reported for the first time in India.

Experimental studies on the vector potential of certain Culex species to West Nile virus.

Experimental studies were carried out to determine the vector potential of four species of mosquitoes to West Nile (WN) virus, viz. Culex tritaeniorhynchus, Cx. vishnui, Cx. bitaeniorhynchus and Cx. univittatus. All the four species of mosquitoes successfully transmitted and supported the growth of WN virus. The study indicated that the four species of mosquitoes could act as potential vectors of WN virus in nature.

Clams as pollution bioindicators in Kuwait's marine environment: metal accumulation and depuration.

This study has developed and verified clam transplantation for marine pollution monitoring in Kuwait and has assessed its reliability for monitoring pollution by heavy metals through laboratory and field transplantation experiments. In field experiments, live specimens of the clam Circenita callipyga were transplanted, either suspended in seawater or buried in sand, to 11 coastal sites in Kuwait; subsamples of transplanted clams were recovered at intervals over a 6-month period. In laboratory experiments, heavy metal accumulation for 36 days and depuration (body's release of pollutants) for 60 days in small and big clams were investigated. Clams subsampled in laboratory and field experiments were analyzed for Hg, Cu, V, Cd and Pb. Results showed the ability of clams to survive under the transplantation conditions. Accumulation of biofouling materials was a problem at only two transplantation sites and was overcome by periodic cleaning. Statistical analysis of laboratory experimental data showed significant accumulation of all test metals and significant depuration of Hg, Cu, V and Cd. Rates of metal accumulation and depuration differed in relation to clam size class. Field experiments indicated statistically significant increases in Hg and Cu concentrations in the transplanted clams at most stations, no change in Cd concentrations and an increase or, occasionally, a decrease in V and Pb concentrations.

Persistence of haemagglutination-inhibition antibodies to JE and WN viruses in naturally infected domestic pigs in Karnataka State, India.

Domestic pigs were used as sentinels to monitor the prevalence of Japanese encephalitis (JE) and West Nile (WN) viruses in Kolar District of Karnataka State, India. The study revealed that the pigs once seroconverted to JE and WN viruses, remained immune as long as they were available for observation, up to three years.

Isolation of Batai virus from sentinel domestic pig from Kolar district in Karnataka State, India.

Batai virus was isolated from the blood sample of one sentinel piglet out of 34 used to monitor the activity of the Japanese encephalitis (JE) virus at Madikere village in Kolar district, Karnataka State, India. This is the first report of the isolation of Batai virus from a mammal.

Experimental studies on the susceptibility of domestic pigs to West Nile virus followed by Japanese encephalitis virus infection and vice versa.

A study on the susceptibility of domestic pigs to West Nile virus (WNV) and Japanese encephalitis virus (JEV) infection was carried out. One batch of pigs was inoculated with WNV followed by JEV and another batch was inoculated vice versa. The first batch developed low level of viraemia and haemagglutination-inhibition (HI) antibodies to both viruses. There was a booster effect on the already existing WNV antibodies after challenging with JEV. In the second batch the animals developed high level of JE viraemia but did not develop WN viraemia. They developed HI antibodies to both JEV and WNV with low booster effect of WNV infection on JEV antibodies. Fresh batches of pigs were infected through bite of WNV- and JEV-infected Culex vishnui mosquitoes. WNV-infected pigs did not show viraemia, whereas JEV-infected ones developed JE viraemia. The study indicated that pigs were poor hosts for WNV but good ones for JEV. However, WNV antibodies reduced the level of JE viraemia and JEV infection boosted the already existing WNV antibodies.

Studies on the mosquito vectors of Japanese encephalitis virus in Mandya District, Karnataka, India.

Entomological investigations were carried out in areas affected by Japanese encephalitis (JE) in Mandya District, Karnataka, India, from 1983 to 1988, to determine species composition and the density of mosquito vectors, in relation to the incidence of JE cases. JE cases occurred in two spells in a year, one during April-June (summer epidemic) and another during October-December (winter epidemic). There was very high incidence of JE cases in extensively irrigated areas and a low incidence in some of the taluks with less or no irrigation systems. Among culicines, Culex tritaeniorhynchus was the most predominant species (20.54%), followed by Cx. fuscocephala (16.94), Cx. vishnui (16.48%), Cx. gelidus (10.70%) and other species. The overall mosquito population showed two peaks in a year, one during the March-April, and another during September, usually preceding the human epidemics. Relative abundance of certain species varied in different years.

Dengue virus and insecticide susceptibility status of Aedes aegypti mosquitoes from Belagola village, Mandya District, Karnataka state: during and post-epidemic investigations.

Aedes aegypti mosquitoes collected during and after dengue virus epidemic season were exposed to diagnostic dosages of insecticides. Mosquitoes collected during the epidemic showed resistance to DDT only, while mosquitoes collected after the control measures showed tolerance to malathion and low resistance to DDT. Since there was no indication of malathion resistance in the mosquitoes collected during the epidemic, the second batch was subjected to malathion selection pressure for 8 generations to find out the mechanism of resistance in this strain. There was 2-4 fold increase in resistance to malathion. Polyacrylamide electrophoresis showed increase in intensity of three esterase bands after selection. Quantitative enzyme assays showed an increase in esterase activity by 3-4 folds. Mosquitoes collected during the epidemic showed higher susceptibility to dengue-2 virus as compared to the laboratory strain, while the mosquitoes collected after malathion fogging showed further increase in the susceptibility to dengue virus. Virus susceptibility of the mosquitoes after eight generations of selection with malathion did not show any further significant increase.

Chemical contamination and toxicity of sediment from a coastal area receiving industrial effluents in Kuwait.

The Shuaiba coastal area (12.5 x 1.5 km) was examined for contamination with total organic carbon, volatile organic matter, total petroleum hydrocarbons, polycyclic aromatic hydrocarbons, cadmium, chromium, copper, lead, nickel, vanadium, and zinc in sediment; their desorption by aqueous elution; and toxicity to aquatic biota. The pollutants were mainly accumulated in the upstream area facing Mina Al-Ahmadi refinery to Shuaiba harbour. Solid-phase Microtox assays showed severe toxicity, and the LC(50) was negatively correlated with most of the chemical parameters, suggesting that toxicity was the function of collective effects of the pollutants present in sediment. Sea water elutriation showed poor desorption of pollutants from sediment, and the elutriates were not found toxic to Microtox and brine shrimp larvae. Whole sediment suspension in sea water reduced the survival of fingerlings in fish bioassays. Action from Shuaiba Area Authority is required to reduce pollutant accumulation in identified depositional area on the Shuaiba coast by facilitating unrestricted water flow in the area and restricting pollutant discharge at source.