Bacillus sp., P. chondroitinus, Herbaspirillum sp., and Photorhabdus luminescens were identified as single unique phylotypes (Table 2, Figure 3). The Good’s coverage calculated for the 85 clones was 68.23% (Table 3). Figure 3 Neighbor-Joining tree deduced from check details partial sequences of 16S rRNA gene Selleck VX-680 clones from field-collected male A. stephensi. Bootstrap confidence values obtained with 1000 resamplings are given at the branch point. Entries with black square represent generic names and accession numbers (in parentheses) from

public databases. Entries from this work are represented as: clone number, generic name and accession number (in parentheses). Table 3 Comparison of the phylotype richness, diversity and evenness values of the isolates and 16S rRNA clones from lab-reared and field-collected A. stephensi mosquitoes. Index Lab-reared A. stephensi PRI-724 Field-collected A. stephensi   Culturable Unculturable Culturable Unculturable   M F M F M F L M F L No. of isolates/clones 18 16 24 24 17 34 30 85 69 66 S a 11 11 15 7 14 29 29 27 36 36 H b 1.74 1.84 2.14 1.97 2.75 2.93 3.21

2.93 3.15 3.49 E c 0.89 0.94 0.89 0.70 0.99 0.93 0.98 0.98 0.98 0.99 C_ACE 45 43 43 31 50 173 157 72 160 123 C_Chao 25 30 30 15 35 104 129 71 117 94 C_Simpson 0.013 0.011 0.08 0.54 0.017 0.02 0.02 0.11 0.11 0.06 Good’s Coverage 39 32 38 71 18 15 13 69 49 46 The table lists the number PJ34 HCl of phylotypes, observed and estimated species richness, coverage and diversity indices for the culturables and 16S rRNA clone libraries from lab-reared and field- collected adult and larval Anopheles stephensi mosquitoes. Numbers were calculated with DOTUR program, OTUs were defined using a distance level of 3%.

The Shannon-Weiner diversity index [16] is calculated as follows: a: S = (Phylotype richness): Total number of species in the sample. b: H = Σ (pi) (log2 p – i), where p represents the proportion of a distinct phylotype relative to the sum of all phylotypes. c: E = (Evenness) was calculated as follows: E = H/Hmax where Hmax = log2 (S) C_ACE = ACE Coverage, C_Chao = Chao Coverage, C_Simpson = Simpson Coverage Good’s Coverage = [1 - (n/N)] × 100 Where n is the number of molecular species represented by one clone (single-clone OTUs) and N is the total number of sequences [54]. M: Adult Male Anopheles stephensi F: Adult Female Anopheles stephensi L: Anopheles stephensi Larvae In all, 64% of the clones were found to belong to firmicutes, followed by 28% from unclassified class of bacteria (mainly uncultured Flexibacteriaceae and uncultured Paenibacillaceae) were also identified. CFB, betaproteobacteria and gammaproteobacteria, each constituted 1% of the total clones (Figure 1). It can be observed here that among culturable isolates gammaproteobacteria are the dominant group, whereas 16S rRNA gene clones were dominated by firmicutes.