Female ARN-509 in vitro Anopheles stephensi A total of 34 distinct isolates were identified from field-collected female A. stephensi midgut microflora. On the basis of phylogenetic

tree 16S rRNA gene sequences were found to belong to major two bacterial phyla, gammaproteobacteria and CFB (Figure 4). The majority of the cultured isolates from field-collected and lab-reared adults belonged to the gammaproteobacteria class. A total of 29 bacterial OTUs were detected among female A. stephensi on the basis of 97% sequence similarity as a cut off value (Table 2). Sequences with more than 97% similarity were considered to be of the same OTUs. Representative genera of gammaproteobacteria were, Acinetobacter sp., A. hemolyticus, A. radioresistens, Citrobacter

freundii, Enterobacter sp., E. cloacae, E. sakazaki, Escherichia hermani and Enterobacteriaceae bacterium. They constituted the largest proportion of 97%, among the total diversity. Out of the 29 distinct phylotypes observed, 28 were found to belong to class gammaproteobacteria only. Only single phylotype Chryseobacterium indologenes, from CFB was detected with 3% proportion from the total observed OTUs. None of the member from high G+C Gram-positive actinobacteria and Gram-positive firmicutes were observed, as in field-collected male A. stephensi. Similarly, none of the CFB group phylotypes were detected in female A. stephensi. Isolates belonging to genus Acinetobacter Chlormezanone sp., A. radioresistens, Enterobacter sp., E. cloacae and Escherichia hermani were commonly observed in both male as well as female field-collected A. stephensi. Selleckchem H 89 These results are quite different from the data what we have observed in lab-reared adult A. stephensi (Figure 1). Figure 4 Phylogenetic tree constructed for partial 16S rRNA gene of isolates cultured from field-collected female A. stephensi. Bootstrap values are given at nodes. Entries with black square represent generic names and accession numbers (in parentheses)

from public databases. Entries from this work are represented as: strain number, generic name and accession number (in parentheses). Female Anopheles stephensi 16S rRNA gene library A total of 100 clones were found positive for the insert and were partially sequenced. Of these, three were shown to be chimeras and were therefore not included for further analysis. The phylogenetic analysis of the remaining clones was done using partial 16S rRNA gene aligned homologous nucleotide sequences (Figure 5). The percentage distribution of the clones from the 16S rRNA gene library representing the microbiota of female A. stephensi midgut was determined (Table 2, Figure 1) On the basis of sequence similarity to the existing GenBank database entries, the clones were clustered together to form four major groups: Gram positive firmicutes, betaproteobacteria and gammaproteobacteria and the unidentified and uncultured bacteria group.