Mosquitoes have evolved an effective innate immune system. The mosquito gut accommodates various microbes, which play a crucial role in shaping the mosquito immune system during evolution. The resident bacteria in the gut microbiota play an essential role in priming basal immunity. In this study, we show that antibacterial immunity in Anopheles gambiae can be enhanced by priming via a sugar meal supplemented with bacteria. Serratia fonticola S1 and Enterobacter sp. Ag1 are gut bacteria in mosquitoes. The intrathoracic injection of the two bacteria can result in an acute hemocoelic infection in the naïve mosquitoes with mortality of ∼40% at 24 h post-infection. However, the Enterobacter orSerratia primed mosquitoes showed a better 24 h survival upon the bacterial challenge. The priming confers the protection with a certain degree of specificity, the Enterobacter primed mosquitoes had a better survival upon the Enterobacter but not Serratia challenge, and the Serratia primed mosquitoes had a better survival upon the Serratia but not Enterobacter challenge. To understand the priming-mediated immune enhancement, the transcriptomes were characterized in the mosquitoes of priming as well as priming plus challenges. The RNA-seq was conducted to profile 10 transcriptomes including three samples of priming conditions (native microbiota, Serratia priming, and Enterobacter priming), six samples of priming plus challenges with the two bacteria, and one sample of injury control. The three priming regimes resulted in distinctive transcriptomic profiles with about 60% of genes affected by both bacteria. Upon challenges, different primed mosquitoes displayed different transcriptomic patterns in response to different bacteria. When a primed cohort was challenged with a heterogenous bacterium, more responsive genes were observed than when challenged with a homogenous bacterium. As expected, many canonical immune genes were responsive to the priming and challenge, but much more non-immune genes with various functions were also responsive in the contexts, which implies that the prior priming triggers a delicately coordinated systemic regulation that results in an enhanced immunity against the subsequent challenge. Besides the participation of typical immune pathways, the transcriptome data suggest the involvement of lysosome and metabolism in the context. Overall, this study demonstrated a trained immunity via priming with bacteria in diet.

With the introduction of molecular taxonomy of mosquitoes, polytene chromosome maps have become indispensable as standard references for locating genes, puffs, and inversion breakpoints of unique DNA sequences. We present a line map and a photomap of the salivary polytene chromosomes of Anopheles (Cellia) subpictus Grassi, an important emerging vector of malaria in India. In addition, we discuss the nature of this species complex consisting of sibling species A, B, C, and D. The comparative study is in relevance to the X chromosome heterozygous inversion differences between 2 allopatric populations of the species and the recognition of 4 X-chromosome inversion genotypes viz: species A-X(+a+b), B-X(ab), C-X(a+b) and D-X(+ab).

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| A molecular analysis of mitochondrial 16s rDNA sequences of hard ticks belonging to genus Hyalomma from Haryana (India) and those available in genbank database was done so as to resolve inter-relationships between members of genus Hyalomma. For this a total of eighty one 16s rDNA sequences belonging to 16 taxa were subjected to molecular and phylogenetic analysis which was conducted in MEGA6 and Beast 1.8.0 software. Phylogenetic relationships were inferred using the bayesian, maximum likelihood (ML) and neighbor joining (NJ) methods by means of Tamura three parameter model + unequal frequency + gamma distribution (TPMuf+G). The analysis revealed 249 variable sites, 232 conserved sites, and 137 parsimony informative sites in the alignment. Results of the phylogenetic analysis provide support for monophyletic origin of genus Hyalomma. Further our results assert that H. anatolicum, H. excavatum, H. marginatum, H. lusitanicum, H. hussaini and H. brevipunctata represent closely related but rapidly diverging taxa and also substantiate H. asiaticum as a species complex. The molecular clock results exemplify divergence time of subfamily Hyalomminae from the common ancestor of subfamily Rhipicephalinae to be 61.99 mya and the origin of family Ixodidae to be 86.79 mya. A basic phylogenetic relationship tree is also provided to help future studies for phylogenetics of family Ixodidae especially for subfamily Hyalomminae.