Evidence of co-infection of chikungunya and densonucleosis viruses in C6/36 cell lines and laboratory infected Aedes aegypti (L.) mosquitoes

<p>Abstract</p> <p>Background</p> <p>Densonucleosis viruses are the etiological agents of insect's disease. We have reported the isolation of densovirus from India and its distribution among the natural populations of <it>Aedes aegypti </it>mosquitoes across the country. Since densonucleosis virus persistently infects mosquito populations, and is demonstrated to negatively affect multiplication of dengue virus in <it>Aedes albopictus</it>, it would be interesting to study if this virus has a role in determining the susceptibility of the vector mosquito <it>Ae. aegypti </it>to chikugunya virus.</p> <p>Methods</p> <p>Mosquito cell lines and adult <it>Ae. aegypti </it>mosquitoes infected with densovirus were superinfected with Chikungunya virus and both the viruses were quantitated by determining their genomic copy number by real time amplification. Comparison was made between the log of genomic copy numbers of the viruses in the presence and absence of each other.</p> <p>Results</p> <p>The log of copy number of the viruses did not vary due to co-infection. Even though the RNA copy number of chikungunya virus increased over the period of time, no change was observed in the RNA copy number between the control and the co-infected group on any given day. Similarly, DNA copy number of densovirus also remained unchanged between the control and the co-infected groups.</p> <p>Conclusion</p> <p>Chikungunya virus neither stimulates the replication of densovirus nor is its own replication suppressed due to co-infection. <it>Ae. aegypti </it>mosquitoes with densovirus infection were as susceptible to infection by chikungunya virus as the uninfected mosquitoes.</p

<i>Drosophila melanogaster</i> as a model host to study arbovirus–vector interaction

ABSTRACTArboviruses cause the most devastating diseases in humans and animals worldwide. Several hundred arbovirus are transmitted by mosquitoes, sand flies or ticks and are responsible for more than million deaths annually. Development of a model system is essential to extrapolate the molecular events occurring during infection in the human and mosquito host. Virus overlay protein binding assay (VOPBA) combined with MALDI TOF/TOF MS revealed that Dengue-2 virus (DENV-2) exploits similar protein molecules in Drosophila melanogaster and Aedes aegypti for its infection. Furthermore, the virus susceptibility studies revealed that DENV-2 could propagate in D. melanogaster, and DENV-2 produced in fruit fly is equally infectious to D. melanogaster and Ae. aegypti. Additionally, real time PCR analysis revealed that RNAi coupled with JAK-STAT and Toll pathway constitutes an effector mechanism to control the DENV-2 infection in flies. These observations point out that D. melanogaster harbors all necessary machineries to support the growth of arboviruses. With the availability of well-established techniques for genetic and developmental manipulations, D. melanogaster, offers itself as the potential model system for the study of arbovirus-vector interactions.</jats:p

Serratia odorifera a midgut inhabitant of Aedes aegypti mosquito enhances its susceptibility to dengue-2 virus.

Mosquito midgut plays a crucial role in its vector susceptibility and pathogen interaction. Identification of the sustainable microflora of the midgut environment can therefore help in evaluating its contribution in mosquito-pathogen interaction and in turn vector competence. To understand the bacterial diversity in the midgut of Aedes aegypti mosquitoes, we conducted a screening study of the gut microbes of these mosquitoes which were either collected from fields or reared in the laboratory "culture-dependent" approach. This work demonstrated that the microbial flora of larvae and adult Ae. aegypti midgut is complex and is dominated by gram negative proteobacteria. Serratia odorifera was found to be stably associated in the midguts of field collected and laboratory reared larvae and adult females. The potential influence of this sustainable gut microbe on DENV-2 susceptibility of this vector was evaluated by co-feeding S. odorifera with DENV-2 to adult Ae. aegypti females (free of gut flora). The observations revealed that the viral susceptibility of these Aedes females enhanced significantly as compared to solely dengue-2 fed and another gut inhabitant, Microbacterium oxydans co-fed females. Based on the results of this study we proposed that the enhancement in the DENV-2 susceptibility of Ae. aegypti females was due to blocking of prohibitin molecule present on the midgut surface of these females by the polypeptide of gut inhabitant S. odorifera

Serratia odorifera a Midgut Inhabitant of Aedes aegypti Mosquito Enhances Its Susceptibility to Dengue-2 Virus

Mosquito midgut plays a crucial role in its vector susceptibility and pathogen interaction. Identification of the sustainable microflora of the midgut environment can therefore help in evaluating its contribution in mosquito-pathogen interaction and in turn vector competence. To understand the bacterial diversity in the midgut of Aedes aegypti mosquitoes, we conducted a screening study of the gut microbes of these mosquitoes which were either collected from fields or reared in the laboratory “culture-dependent” approach. This work demonstrated that the microbial flora of larvae and adult Ae. aegypti midgut is complex and is dominated by Gram negative proteobacteria. Serratia odorifera was found to be stably associated in the midguts of field collected and laboratory reared larvae and adult females. The potential influence of this sustainable gut microbe on DENV-2 susceptibility of this vector was evaluated by co-feeding S. odorifera with DENV-2 to adult Ae. aegypti females (free of gut flora). The observations revealed that the viral susceptibility of these Aedes females enhanced significantly as compared to solely dengue-2 fed and another gut inhabitant, Microbacterium oxydans co-fed females. Based on the results of this study we proposed that the enhancement in the DENV-2 susceptibility of Ae. aegypti females was due to blocking of prohibitin molecule present on the midgut surface of these females by the polypeptide of gut inhabitant S. odorifera

Relative percent distribution of microbes in fourth <i>in star</i> larval midgut and their surrounding habitat.

*<p>Identification of isolates is based on conventional biochemical characterization and 16s rDNA sequencing.</p>%<p>Relative distribution = CFU of a particular type of microbe/CFU of all types of microbes in given sample/niche ×100.</p