Interaction of azadirachtin with the lipid-binding domain: Suppression of lipid transportation in the silkworm, Bombyx mori.

This study investigates the effects of the insect growth regulator azadirachtin on lipid transportation to the ovary of the silkworm, Bombyx mori. Lipids are hydrophobic in nature and require a carrier for circulation in the blood. Protein-lipid interactions play a vital role in lipid transport, thereby keeping the system balanced. In general, lipids bind to lipoproteins in a specific region called the lipid-binding domain (LBD). In this study, B. mori apolipophorin amino acid sequences were retrieved from NCBI and the LBD was identified. The LBD structure was predicted by (PS)2 and validated in ProSA. The LBD structure was docked with DMPC, POPC and sphingomyelin by SwissDock, each binding with GLN 171, ASN 162, and ASN 160 and 162, respectively. Interestingly, azadirachtin binds with ASN 160 and 162 and GLN 171, which shows that lipids and azadirachtin are binding with the same amino acid residues in the LBD. Later, this result was confirmed with wet lab work using a fluorescent phospholipid probe. Azadirachtin binding with the LBD was indirectly proportional to the fluorescent lipid binding. These results suggest that azadirachtin binds with the LBD instead of the lipids and interrupts the protein-lipid interaction, leading to the suppression of lipid transportation to the ovary.

Vector ecology of human schistosomiasis in south India and description of a new species of the genus Ferrissia (Mollusca: Gastropoda: Planorbidae).

BACKGROUND & OBJECTIVES: Vector ecology and taxonomy of snails is a prerequisite for controlling schistosomiasis in the tropics. The ecology of the freshwater limpet genus Ferrissia was investigated for detection of cercariae larvae in them, and taxonomic description of a new species of the genus Ferrissia. METHODS: This study was conducted in 15 perennial streams from five different hills of south India. To study the seasonal patterns, a stream from each hill was selected and sampled in three seasons. In each study site, triplicate sampling was done and specimens were collected from stream substrates as well as waste material submerged in stream. Microscopic examination was carried out for detecting cercariae larvae in limpets. RESULTS: Three freshwater limpets (F. tenuis, F. verruca and F. fivefallsiensis) were observed. Seasonality influenced the abundance of limpets. The highest abundance was observed during post-monsoon (December and January). The distribution of Ferrissia was observed at riffle in pebbles, leaf litter and wastes (polyethylene bags and snacks cover) submerged in water. No cercariae larvae were found from the body of limpets. In this study, we described a new species of Ferrissia fivefallsiensis. INTERPRETATION & CONCLUSION: Our results showed the distribution, habitat preference and seasonality of limpets, and recommend the detection of Schistosoma from limpets as well as human samples by use of molecular tools.

Pheromone-binding proteins based phylogenetics and phylogeography of Maruca spp. from Asia, Africa, Oceania, and South America.

Variations in the functional response of legume pod borer (Maruca vitrata) populations to sex pheromone blends were observed in Asia and Africa. Hence, this study was carried out to understand the differences in pheromone-binding proteins (PBPs) among Maruca populations in Asia, Africa, Oceania, and South America. A de novo transcriptome assembly was adopted to sequence the entire transcribed mRNAs in M. vitrata from Taiwan. The raw-sequence data were assembled using homologous genes from related organisms in GenBank to detect M. vitrata PBPs (MvitPBPs). Sections of the cDNA of MvitPBP of different length were used to design primers to amplify the full-length cDNA of PBPs. All three PBP sequences comprised three exons interspersed by two introns. In total, 92 MvitPBP1 haplotypes, 77 MvitPBP2 haplotypes, and 64 MvitPBP3 haplotypes were identified in 105, 98, and 68 Maruca individuals, respectively. High pairwise F ST values (0.41-0.73) and phylogenetic analyses distinguished the putative Maruca species in South America from those occurring in rest of the world, and possibly two putative subspecies in Asia and Africa. The haplotype networks and Automatic Barcode Gap Discovery analyses also confirmed these results. The negative Tajima's D and Fu's F S values showed the recent demographic expansion of Maruca populations. Thus, this study confirmed the presence of different Maruca species and/or subspecies in different continents based on the diversity within PBP genes. Additional sampling and studies are suggested for Oceania and South America. The genetic differences among Maruca populations should be carefully considered while using sex pheromone lures and bio-control agents.

Hierarchical dynamics influence the distribution of immature black flies (Diptera: Simuliidae).

Adult black flies (Simuliidae) are medically important insects and they are the sole vector of Onchocerca volvulus. Immature black flies are major components of aquatic macroinvertebrate assemblages in streams and play a vital role in nutrient dynamics. In this study, we examined effect of hierarchical dynamics (spatio-temporal pattern) on the distribution of immature black flies in South Indian streams. The sampling was done in streams of Western Ghats, South India. A total of 16 species belong to two subgenera: Simulium (10 species) and Gowmphostilbia (6 species) of Simulium were observed. Alpha diversity indices were analyzed, which indicate the abundance and species richness between sampling sites. Non-parametric analysis recognized the key environmental variables including latitude and stream order. Subsequently, the monsoon influences the larval assemblages and its association was high in leaf litter as revealed through statistical analyses. Although the members of the immature black fly assemblage with different environmental factors, they are very closely related to spatial and temporal organization and secondarily with other factors prevailing in streams.

Phylogeographical Structure in Mitochondrial DNA of Legume Pod Borer (Maruca vitrata) Population in Tropical Asia and Sub-Saharan Africa.

This study was undertaken to assess the genetic diversity and host plant races of M. vitrata population in South and Southeast Asia and sub-Saharan Africa. The cytochrome c oxidase subunit 1 (cox1) gene was used to understand the phylogenetic relationship of geographically different M. vitrata population, but previous studies did not include population from Southeast Asia, the probable center of origin for Maruca, and from east Africa. Extensive sampling was done from different host plant species in target countries. Reference populations from Oceania and Latin America were used. An amplicon of 658 bp was produced by polymerase chain reaction, and 64 haplotypes were identified in 686 M. vitrata individuals. Phylogenetic analysis showed no difference among the M. vitrata population from different host plants. However, the results suggested that M. vitrata has formed two putative subspecies (which cannot be differentiated based on morphological characters) in Asia and sub-Saharan Africa, as indicated by the high pairwise FST values (0.44-0.85). The extremely high FST values (≥ 0.93) of Maruca population in Latin America and Oceania compared to Asian and African population seem to indicate a different species. On the continental or larger geographical region basis, the genetic differentiation is significantly correlated with the geographical distance. In addition, two putative species of Maruca, including M. vitrata occur in Australia, Indonesia and Papua New Guinea. The negative Tajima's D and Fu's FS values showed the recent demographic expansion of Maruca population. The haplotype network and Automatic Barcode Gap Discovery analyses confirmed the results of phylogenetic analysis. Thus, this study confirmed the presence of three putative Maruca species, including one in Latin America, one in Oceania (including Indonesia) and M. vitrata in Asia, Africa and Oceania. Hence, the genetic differences in Maruca population should be carefully considered while designing the pest management strategies in different regions.

TBT effects on the development of intersex (ovotestis) in female fresh water prawn Macrobrachium rosenbergii.

The impact of tributyltin (TBT) on the female gonad and the endocrine system in Macrobrachium rosenbergii was studied. Prawns were exposed to environmentally realistic concentrations of 10, 100, and 1000 ng/L of TBT for 6 months. Dose dependent effects were noticed in TBT exposed prawns. At 1000 ng/L TBT caused ovotestis formation (formation of male germ cells in ovary). Presence immature oocytes, fusion of developing oocytes, increase in interstitial connective tissues, and its modification into tubular like structure and abundance of spermatogonia in the ovary of TBT treated prawns. The control prawn ovary showed normal architecture of cellular organelles such as mature oocytes with type 2 yolk globules, lipid droplets, normal appearance of yolk envelop, and uniformly arranged microvilli. On the other hand, type 1 yolk globules, reduced size of microvilli, spermatogonial cells in ovary, spermatogonia with centrally located nucleus, and chromatin distribution throughout the nucleoplasm were present in the TBT treated group. Immunofluorescence staining indicated a reduction in vitellin content in ovary of TBT treated prawn. Moreover, TBT had inhibited the vitellogenesis by causing hormonal imbalance in M. rosenbergii. Thus, the present investigation demonstrates that TBT substantially affects sexual differentiation and gonadal development in M. rosenbergii.

Insect gut microbiome - An unexploited reserve for biotechnological application

Metagenomics research has been developed over the past decade to elucidate the genomes of the uncultured microorganisms with an aim of understanding microbial ecology. On the other hand, it has also been provoked by the increasing biotechnological demands for novel enzymes, antibiotic and signal mimics. The gut microbiota of insects plays crucial roles in the growth, development and environmental adaptation to the host insects. Very recently, the insect microbiota and their genomes (microbiome), isolated from insects were recognized as a major genetic resources for bio-processing industry. Consequently, the exploitation of insect gut microbiome using metagenomic approaches will enable us to find novel biocatalysts and to develop innovative strategies for identifying smart molecules for biotechnological applications. In this review, we discuss the critical footstep in extraction and purification of metagenomic DNA from insect gut, construction of metagenomic libraries and screening procedure for novel gene identification. Recent innovations and potential applications in bioprocess industries are highlighted.