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BackgroundMalaria is a major cause of morbidity and mortality in Northeast India. As there is limited information available on the potential influence of socio-economic variables on malaria risk, the present study was conducted to assess the influence of demographic factors, the socio-economic status, and knowledge, awareness and education on malaria occurrence.MethodsDemographics, malaria knowledge and socio-economic variables were collected in four randomly selected health sub-centres of the Orang primary health centre in the Udalguri district, Assam and the association of malaria occurrence with different variables were analysed. The trend of malaria occurrence for different income groups, proximity to health centres and number of mosquito bites per day was also determined using the chi-square test. Relative risk (RR) for gender, house type, knowledge and use of bed nets was determined using Katz approximation.ResultsOut of the 71 household heads interviewed, 70.4% (50/71) were males. About half (54.9%, 39/71) of the participants had a history of malaria in the last two years, of which 64.1% (25/39) were males, while 35.9% (14/39) were females (χ2 = 5.13; p = 0.02; RR = 1.79). Of the total population surveyed, 49.3% lived in bamboo houses and 35.2% lived at a distance of >3 km from the nearest health centre. The number of participants who had a history of malaria decreased with an increasing monthly income (p < 0.0001). Malaria occurrence was higher among the households living in bamboo houses (69.2%), as compared to Kucha houses (20.5%) and Pucca houses (10.3%). No significant association was observed between education level and malaria occurrence (p = 0.93). The participants who did not use bed nets regularly reported a high occurrence of malaria infection as compared to those who used bed nets everyday (p < 0.0001).ConclusionsLower income, house type, distance to health sub-centre, knowledge and awareness about malaria, number of mosquito bites per day and use of bed nets were positively associated with malaria occurrence. Increasing the number of health sub-centres close to rural areas, improving the economic status and increasing awareness about malaria prevention measures will thus help to reduce malaria-associated morbidities.

BackgroundMicrobiota inhabiting midguts of mosquitoes play a key role in the host - parasite interaction and enhance vectorial capacity of viral diseases like dengue and chikungunya fevers. Mosquito midgut is considered to be an important site for host-pathogen interaction and pathogen survival is thought to be an outcome of this interaction. In the present study we examined the bacterial community in the midgut of Aedes mosquitoes in Arunanchal Pradesh, India, a subtropical zone where dengue fever is reported to be emerging.MethodLarvae and pupa of Aedes mosquitoes were collected from a biodiversity hotspot, Bhalukpong, Arunachal Pradesh, India. 16S rRNA gene sequences were used for identification of isolated bacterial population from each species of mosquitoes. We used various diversity indices to assess the diversity and richness of the bacterial isolates in both mosquito species.ResultOn the basis of 16S rRNA gene sequence analysis a total of 24 bacterial species from 13 genera were identified belonging to 10 families of four major phyla. Phylum Proteobacteria was dominant followed by Firmicutes, Bacteroidetes and Actinobacteria. The midgut bacteria belonging to the phylum Proteobacteria and Firmicutes were isolated from both Ae. albopictus and Ae. aegypti, whereas, bacteria belonging to phylum Bacteroidetes and Actinobacteria were isolated only from Ae. albopictus and Ae. aegypti respectively. Enterobacter cloacae was the dominant bacterial species in both Ae. albopictus (33.65 %) and Ae. aegypti (56.45 %). Bacillus aryabhattai (22.78 %) was the second most common bacterial species in Ae. albopictus whereas, in Ae. aegypti the second most common bacterial species was Stenotrophomonas maltophilia (7.44 %).ConclusionThe family Enterobacteriaceae of phylum Proteobacteria was dominant in both species of Aedes mosquitoes. To the best of our knowledge, this is the first attempt to study midgut microbiota from a biodiversity hotspot in Northeastern India. Some bacterial genera Enterobacter and Acinetobacter isolated in this study are known to play important roles in parasite-vector interaction. Information on midgut microflora may lead towards the development of novel, safe, and effective strategies to manipulate the vectorial capacity of mosquitoes.

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The mosquito Culex quinquefasciatus is a ubiquitous species that serves as a major vector for west nile virus and lymphatic filariasis. Ingestion of bloodmeal by females triggers a series of physiological processes in the midgut and also exposes them to infection by these pathogens. The bacteria normally harbored in the midgut are known to influence physiology and can also alter the response to various pathogens. The midgut bacteria in female Cx. quinquefasciatus mosquitoes collected over a large geographical area from India was studied. Examination of 16S ribosomal DNA amplicons from culturable microflora revealed the presence of 83 bacterial species belonging to 31 bacterial genera. All of these species belong to three phyla i.e. Proteobacteria, Firmicutes and Actinobacteria. Phylum Proteobacteria was the most dominant phylum (37 species), followed by Firmicutes (33 species) and Actinobacteria (13 species). Phylum Proteobacteria, was dominated by members of γ-proteobacteria class. The genus Staphylococcus was the largest genus represented by 11 species whereas Enterobacter was the most prevalent genus and recovered from all the field stations except Leh. Highest bacterial prevalence was observed from Bhuj (22 species) followed by Nagrota (18 species), Masimpur (18 species) and Hathigarh (16 species). Whereas, least species were observed from Leh (8 species). It has been observed that individual mosquito harbor extremely diverse gut bacteria and have very small overlap bacterial taxa in their gut. This variation in midgut microbiota may be one of the factors responsible for variation in disease transmission rates or vector competence within mosquito population. The present data strongly encourage further investigations to verify the potential role of the detected bacteria in mosquito for the transmission of lymphatic filariasis and west nile virus. To the best of our knowledge this is the first study on midgut microbiota of wild Cx. quinquefasciatus from over a large geographical area.

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Viruses are a cause of significant health problem worldwide, especially in the developing nations. Due to different anthropological activities, human populations are exposed to different viral pathogens, many of which emerge as outbreaks. In such situations, discovery of novel viruses is utmost important for deciding prevention and treatment strategies. Since last century, a number of different virus discovery methods, based on cell culture inoculation, sequence-independent PCR have been used for identification of a variety of viruses. However, the recent emergence and commercial availability of next-generation sequencers (NGS) has entirely changed the field of virus discovery. These massively parallel sequencing platforms can sequence a mixture of genetic materials from a very heterogeneous mix, with high sensitivity. Moreover, these platforms work in a sequence-independent manner, making them ideal tools for virus discovery. However, for their application in clinics, sample preparation or enrichment is necessary to detect low abundance virus populations. A number of techniques have also been developed for enrichment or viral nucleic acids. In this manuscript, we review the evolution of sequencing; NGS technologies available today as well as widely used virus enrichment technologies. We also discuss the challenges associated with their applications in the clinical virus discovery.

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