Seroprevalence of dengue in a rural and an urbanized village: A pilot study from rural western India.

BACKGROUND & OBJECTIVES: Dengue is highly prevalent in tropical and subtropical regions. The prevalence of dengue is influenced by number of factors, i.e. host, vector, virus and environmental conditions including urbanization and population density. A cross sectional study was undertaken to determine the seroprevalence of dengue in two selected villages that differed in the level of their urbanization and population density. METHODS: Two villages with demographically well-defined populations close to Pune, a metropolitan city of western India, were selected for the study. Age stratified serosurvey was carried out during February to May 2011 in the two villages-a rural village A, located 6 km from the national highway with a population density of 159/km2 ; and an urbanized village B, located along the highway with a population density of 779/km2 . Assuming a low seroposi- tivity of 10%, 702 serum samples were collected from village A. Sample size for village B was calculated on the basis of seropositivity obtained in village A, and 153 samples were collected. Serum samples were tested for the presence of dengue virus (DENV)-specific IgG. Simple proportional analyses were used to calculate and compare the seroprevalence. RESULTS: Of the 702 samples collected from village A, 42.8% were found positive for anti-DENV IgG. A significantly higher seropositivity for DENV (58.8%) was found in village B. In village A, there was an age dependent increase in seroprevalence; whereas, in village B, there was a steep increase from 17% positivity in 0-10 yr age group to 72% in the 11-20 yr age group. The seroprevalence was almost similar in the older age groups. INTERPRETATION & CONCLUSION: The observations suggested that prevalence of dengue is probably associated with urbanization and host population density. Areas that are in the process of urbanization needs to be monitored for prevalence of dengue and its vector, and appropriate vector control measures may be implemented.

Hepatitis B virus: predominance of genotype D in primitive tribes of the Andaman and Nicobar islands, India (1989-1999).

To understand the possible origin of hepatitis B virus (HBV), three of the four hyperendemic, primitive accessible tribes of the Andaman and Nicobar islands, India, were investigated. The Nicobarese tribe was investigated in 1989 and 1999. The S gene from 65 HBV isolates was amplified by PCR and sequenced. Genotyping and serotyping were carried out on the basis of phylogenetic and amino acid analyses of S gene. All 20 Nicobarese-89 isolates, nine Onges-99 isolates and the single Andamanese-99 HBV isolate were classified as genotype D. Of the Nicobarese-99 isolates, 32 (91.4 %) and three (8.6 %) were genotypes D and A, respectively. Per cent nucleotide identity between the S sequences representing different tribes varied from 98.06 to 98.59 % and varied from mainland isolates by 1.6-2.0 %. Although southeast Asian origin is postulated for the Nicobarese tribe, the presence of different genotypes suggests introduction of HBV after migration to these islands, probably from mainland India, 200 years back, when these islands became inhabited as a part of penal settlement during the British regimen.

Characterization of measles sequences from Pune, India.

The goal of this study was to conduct the initial genetic characterization wild-type measles viruses currently circulating in India. Reverse transcription-polymerase chain reaction detected measles RNA in 11 of 14 throat swabs collected from sporadic and outbreak-associated cases in the city of Pune, during 1996-1998. Sequence analysis of the H and N genes showed that six sequences were genotype D4, three were genotype D8, and two were genotype A. Continued virologic surveillance in other areas of India as well as neighboring countries will indicate the extent of genetic diversity present among wild-type measles viruses circulating in India.

A serological survey of arboviral diseases among the human population of the Andaman and Nicobar Islands, India.

In an attempt to determine the prevalence of certain arthropod-borne viruses of public health importance amongst the human population of the Andaman and Nicobar Islands of India, 2,401 sera were collected from six major localities. The sera were analysed by the hemagglutination inhibition (HI) and neutralization (N) tests, using Chikungunya (CHIK), Japanese encephalitis (JE), West Nile (WN), dengue (DEN-2), Langat (TP-21) and Kyasanur Forest disease (KFD) viral antigens. The highest prevalence of HI antibodies was detected against KFD virus (22.4%), followed by Langat (20.2%), JE (5.9%), DEN-2 (3.1%), CHIK (2.9%) and WN (0.8%) viruses. Cross-reactions to the viral antigens were also noted. The results of N tests indicated a high prevalence of DEN-2 (25.4%) virus, followed by Langat (17.5%), CHIK (15.3%), KFD (12%), JE (2.19%) and WN (1.8%). These results are discussed in relation to important epidemiological parameters like age, sex and geographical location. To our knowledge, this is the first report of an extensive serosurvey of arthropod-borne viruses on these islands.

Prevalence of West Nile virus infection in India.

During the course of the virological investigation of cases of suspected viral fevers carried out at the National Institute of Virology (NIV), Pune, India, evidence of recent infection with West Nile (WN) virus was detected in 88 cases. Fever, general aches, headache, nausea and vomiting were the principal clinical features in 92% (81/88) of the cases; there were seven cases of encephalitis, in which WN virus-specific IgM class antibodies were detected in CSF samples. These cases of encephalitis were from Japanese encephalitis (JE) nonendemic areas, like Maharashtra and Rajasthan, as well as from JE endemic areas, like Goa and Orissa. Interestingly, neutralizing antibodies predominantly to WN virus were detected in CSF samples by the 50% cytopathic effect inhibition method; the titers ranged from 5 to 375. Cases of WN virus infection associated with both encephalitis and classic features have been reported for the first time in recent years in India. Reports of unique urban West Nile virus encephalitis epidemics in New York, Romania, and Algeria in recent years have signaled the emergence of neurological infection due to West Nile virus as a novel public health threat. This study is important because it records evidence of WN virus infection in India.

Horizontal and vertical transmission of dengue virus type 2 in highly and lowly susceptible strains of Aedes aegypti mosquitoes.

Isofemale lines of Aedes aegypti mosquitoes highly and lowly susceptible to dengue type 2 (DEN-2) virus (DEN(h) and DEN(l), respectively) were established by oral feeding and individual rearing. The susceptibility at F13 generation was found to be 61% and 25% for the DEN(h) and DEN(l) line, respectively. The virus-infected mosquito females were allowed to probe on bovine albumin phosphate saline pH 7.2 (BAPS) through membrane feeders. The presence of virus in the probed BAPS was determined either by ELISA or by intrathoracic (i.t.) inoculation of mosquitoes or by both methods. The rate of oral transmission of virus was found to be 2 times higher in the DEN(h) isofemale line than in the DEN(l) one. Similarly, vertical transmission rate of the virus was found to be 7 times higher in the DEN(h) line. When batches of eggs obtained from infected female mosquitoes were allowed to hatch after two months the vertical transmission rate of the virus was very high. It is possible that, at room temperature, the virus gets an opportunity to multiply and increase its copy number in the quiescent embryos. The progeny obtained from the infected mosquitoes was found to be capable of transmitting the virus horizontally when allowed to probe on BAPS through the membrane feeder. This is the first report demonstrating horizontal transmission of DEN-2 virus by mosquitoes infected through vertical transmission. The higher vertical transmission rate of the virus in the progeny obtained from the eggs dessicated for a longer time and the horizontal transmission of the virus from the progeny is of very high epidemiological significance.

Prevalence of anti-hepatitis E virus antibodies in different Indian animal species.

Prevalence of IgG antibodies to hepatitis E virus (IgG-anti-HEV) was determined among different animal species from India. Seropositivity varied from 4.4% to 6.9% in cattle, 54.6-74.4% in pigs and 2.1-21.5% in rodents. Of the 44 dogs screened, 10 were positive (22.7%). None of the 250 goat sera tested were found to be anti-HEV positive. Among rodents, over 50% serum samples collected in 1985 from Bandicota bengalensis were positive for anti-HEV antibodies. No evidence of HEV infection was obtained following experimental inoculation of an Indian strain (AKL-90) of HEV into anti-HEV negative pigs and goats. The results document varied prevalence of anti-HEV antibodies in different animal species from India and of inability of Indian pigs and goats to support replication of at least one human strain of HEV.

An outbreak of dengue fever in rural areas of northern India.

During the past few decades, epidemics of dengue fever are causing concern in several South-East Asian countries including India. The rural areas of Hissar district of Haryana state, situated about 170 km North-West of Delhi, experienced an outbreak of febrile illness during July-August 1996. A total of 13 villages in eight affected primary health centres reported fever cases. The clinical, epidemiological and entomological findings indicated that the present episode was due to dengue fever. The aetiological agent of the current outbreak, the DEN-2 virus, was isolated from 12 acute-phase sera specimens. Though, in the recent past outbreaks have been reported from the rural areas of southern and western India, the present episode is the first outbreak being reported from the rural areas of northern India. The increasing frequency of dengue fever outbreaks in rural areas of various Indian states reflects the changing life style of the rural population as a result of urbanization process and calls for a suitable prevention and control policy based on strengthened surveillance, appropriate health education to the community coupled with proper training of health personnel.

Isolation of chikungunya virus from Aedes aegypti mosquitoes collected in the town of Yawat, Pune District, Maharashtra State, India.

Chikungunya (CHIK) virus is prevalent throughout Southeast Asia and Africa. It has caused numerous large outbreaks in India. No active or passive surveillance has been carried out since the last epidemic occurring in 1971. During a recent outbreak of Dengue (DEN)-like illness in eastern India, Aedes aegypti mosquitoes collected from the affected area were positive for CHIK virus. Evidence of dual infection with CHIK and DEN typel virus was also obtained. A widely circulating low-virulent CHIK virus is a possible explanation for the epidemiological pattern of the CHIK virus disease in this region.

Japanese encephalitis in Sangli district, Maharashtra.

During the months June to December, 1997, 52 cases of suspected viral encephalitis were admitted at the Government Hospital, Sangli. These cases were from the congested areas of Sangli and the adjoining villages. All age groups and both genders were affected. IgM antibodies to Japanese encephalitis (JE) virus were detected in the sera of five of the 52 cases. Perhaps JE virus has established itself in a new locality in Maharashtra and could cause serious public health problems.

Putative chikungunya virus-specific receptor proteins on the midgut brush border membrane of Aedes aegypti mosquito.

Two proteins (putative receptors) of 60 and 38 kDa, for chikungunya (CHIK) virus were detected in the brush border membrane fraction (BBMF) of the normal population of Aedes aegypti mosquitoes. Mosquitoes were infected orally with CHIK virus and infectivity checked by testing the head squashes. BBMF was prepared from proved positive and negative mosquitoes. The receptor proteins were found to be present in both the proved genotypes. However, dot-b'ot assays showed that the CHIK virus binding activity of BBMF/mg protein was noticeably low in the proved negative mosquitoes as compared to the positives. BBMF from the larvae of the normal populations also showed the presence of the receptor proteins, binding to CHIK virus. Receptor proteins from larvae as well as the adults were found glycosylated. CHIK virus receptor proteins of 24, 45, 58, 60 and 62 kDa were also seen in the membrane fraction of the C6/36 cells.

A retrospective serological survey of humans in India for Wanowrie virus.

Wanowrie virus (WAN) is an ungrouped arbovirus having morphogenetic characters in common with Bunyavirus. It was isolated for the first time from ticks, Hyalomma marginatum isacci (6700 strain) and subsequently from a human case (68923 strain). In the absence of any reports on the prevalence of this virus amongst humans in India, 600 human sera were tested in the Complement Fixation (CF) test against both the strains of WAN virus. One hundred human sera were randomly selected from six different states, viz Jammu and Kashmir, Rajasthan, Orissa, Maharashtra, Karnataka and Tamil Nadu. CF antibodies were detected in 77 sera (12.8%) against the 6700 antigen while 91 sera (15.1%) possessed antibodies against the 68923 antigen; 60 sera were found to be reactive against both the antigens, with almost identical titres. There was no statistical difference in the distribution of CF reactive sera with respect to age, gender and geographic locations. All the CF reactive sera were found to be negative when tested in the neutralization test against the respective virus strains. The CF antibodies detected in these sera may have been produced during exposure to an antigenically closely related virus yet to be identified.

Investigation of buffalopox outbreaks in Maharashtra State during 1992-1996.

During 1992-96, outbreaks of buffalopox zoonosis were reported from different villages in Jalgaon, Dhule and Beed districts of Maharashtra State. In humans, pox lesions were observed on the hands whereas in affected buffaloes and cows the lesions were noticed mainly on the teats and udder. Twenty two virus strains were isolated from the skin scabs collected from infected humans and milch animals. Neutralizing antibodies were detected not only in the sera of affected humans but also in their contacts. Detection of antibodies in young individuals from endemic area, who were neither vaccinated for smallpox nor had any contact with buffaloes or history of any poxvirus disease, is suggestive of occurrence of subclinical infection. A few children who had no contact with infected animals also showed clinical manifestations with disseminated lesions on the face, arm and buttocks, and thus suspected to have acquired infection through their infected parents or other family members indicating a possible man to man transmission. Therefore, in the light of discontinuation of smallpox vaccination, buffalopox outbreaks need to be monitored carefully as this may emerge as a serious zoonotic disease in India.

The 1993 epidemic of dengue fever in Mangalore, Karnataka state, India.

An epidemic of febrile illness with hemorrhagic manifestations occurred in certain parts of Mangalore city, Karnataka state, India, from the last week of July 1993. The epidemic reached its peak by mid-August and then started declining. Sporadic cases, however, continued to occur till early December. About 200 cases were reported covering all age groups and both sexes. The cases presented with pyrexia, myalgia, arthralgia and headache. Palatal petechiae, magenta colored tongue with central coating, maculopapular rash and facial flush were observed as classical signs. The tourniquet test was positive in 12% of the cases. Hemorrhage was observed in the form of epistaxis (2 cases), subconjunctival hemorrhage (2 cases) or purpura (3 cases). There were no deaths which were attributable to the epidemic. Five strains of dengue (DEN-2) virus were recovered from the acute-phase sera. Dengue virus-specific IgM type of antibodies were detected in 29/116 (25%) sera. Breeding of Aedes aegypti was observed in some of the areas where cases had occurred. No virus was isolated from any of the field-caught Ae. aegypti mosquitos.