Vinod Joshi

Importance of Socioeconomic Status and Tree Holes in Distribution of Aedes Mosquitoes (Diptera: Culicidae) in Jodhpur, Rajasthan, India

Abstract Immature Aedes mosquitoes were found in domestic, peridomestic, and tree hole habitats within three socioeconomic strata of Jodhpur, a city within an arid area of Rajasthan, India, endemic for dengue. Peridomestic habitats served as a persistent source of Aedes vectors, especially those used for watering cows for religious reasons that were located within high socioeconomic areas. Domestic (indoor) containers within low socioeconomic strata showed a higher container index (27.0%) than periurban areas with cattle sheds (14.3%) or high socioeconomic areas (18.1%). Mosquitoes were collected in tree holes at zoos and gardens supporting several species of monkeys. Six of 67 Aedes albopictus Skuse reared from immatures collected in tree holes tested positive for dengue antigen acquired through vertical transmission, possibly indicating a persistence mechanism for dengue virus within an urban environment.

Rare occurrence of natural transovarial transmission of dengue virus and elimination of infected foci as a possible intervention method

Transovarial transmission of dengue virus has been studied in 33 districts of Rajasthan, India. Small proportion (1.09%) of breeding containers positive for the virus and their elimination has been demonstrated as a possible intervention method of disease control. Dengue virus was isolated from individual mosquitoes employing Indirect Fluorescence Antibody Test and Reverse Transcriptase Polymerase Chain Reaction. Out of 1,30,525 containers examined only 1432(1.09%) showed transovarially transmitted virus activity. Elimination of larvae from all the 1432 virus positive containers resulted in substantial control over prospective transmission of dengue. The study highlights rarity of transovarial transmission under natural conditions and sensitizes whether elimination of vertically infected foci could be used as a new intervention method.

Multiple dengue virus types harbored by individual mosquitoes.

The existing knowledge on pathogenesis and aetiology of DHF establishes that Dengue Hemorrhagic Fever (DHF) and Dengue Shock Syndrome (DSS) are caused by two subsequent infections of two different serotypes of dengue affecting a common human population with a time gap. Present studies have been undertaken on 212 laboratory reared infected individual mosquitoes from larvae collected from 31 dengue endemic towns of Rajasthan, India. Type specific DEN viruses were detected from individual mosquitoes employing RT-PCR. In 78.7% of 212 infected individual mosquitoes studied, vertically transmitted multiple DENV types were observed. We report for the first time that single mosquitoes contain multiple dengue virus types.

In-silico Designing and Testing of Primers for Sanger Genome Sequencing of Dengue Virus Types of Asian Origin

Rarity in reporting whole genome sequence of Dengue virus from dengue endemic countries leaves lacunae in understanding regional pattern of virus mutation and ultimately leading to non-understanding of transmission pattern and clinical outcomes emerging at regional levels. Due to inter-serotype genomic similarity and intra-serotype genomic diversity, appropriate designing of primer pairs appears as an exhaustive exercise. Present paper reports new Dengue virus type-specific primer which may help in characterizing virus specific to Asian origin. Genomes of dengue virus serotypes of Asian region were searched and using advanced bioinformatics tools, serotype specific primers were designed and tested for their targeted amplification efficiency. 19 primers sets for DENV-1, 18 primer sets for DENV-2, 17 for DENV-3 and 18 for DENV-4 were designed. In-silico and experimental testing of the designed primers were performed on virus isolated from both clinical isolates and passaged cultures. While all 17 and 18 primer sets of DENV-3 and DENV-2 respectively yielded good quality sequencing results; in case of DENV-4, 16 out of 18 primer sets and in DENV-1, 16 out of 19 primer sets yielded good results. Average sequencing read length was 382 bases and around 82% nucleotide bases were Phred quality QV20 bases (representing an accuracy of circa one miscall every 100 bases) or higher. Results also highlighted importance of use of primer development algorithm and identified genomic regions which are conservative, yet specific for developing primers to achieve efficiency and specificity during experiments.