Characterization of complete mitochondrial genome of three Horse flies of the genus Tabanus (Diptera: Tabanidae): comparative analysis.

BACKGROUND: Tabanidae (Horse-Flies or Deer-Flies) are one of the most economically important as well as medically significant haematophagous insect family within the order Diptera. Members of this group are also responsible for the mortality of substantial number of live-stock every year. Due to their pathogen transmission potential and vector competencies makes them an important insect group to study. Till now, mitochondrial genome of 18 species of tabanids were available. METHODS AND RESULTS: The complete mitogenome of three species T. diversifrons (15,809 bp), T. rubidus (15,878 bp) and T. tenens (15,872 bp) were generated by Next generation sequencing method. They consist 37 genes, with a positive AT skew and a negative GC skew. The gene order of these three species is similar to the typical gene arrangement of infra-order Tabanomorpha. Most of the tRNAs showed typical clover-leaf secondary structure except trnS1, which lacks the DHU arm. The sliding window analysis showed that the nad4L is the most conserved while atp8, and nad6 are the most variable genes. Moreover, the ratios of non-synonymous to synonymous substitution rates indicated that all PCGs under the purifying selection. Phylogeny revealed Chrysops and Haematopota are monophyletic while species of Hybomitra are nested within the polyphyletic clade of Tabanus. T. diversifrons exhibits sister relationship with Atylotus miser. Two morphologically divergent species T. rubidus and T. tenens are found to be genetically similar and indistinguishable by mitochondrial genome. CONCLUSIONS: The hypervariable genes like atp8 and nad6 can be used as molecular markers for the identification of recently diverged lineages of family Tabanidae. Further, to address uncertainties arising from the two morphological divergent species, it is imperative to obtain data from nuclear gene markers.

The Sela macaque (Macaca selai) is a distinct phylogenetic species that evolved from the Arunachal macaque following allopatric speciation.

Arunachal macaque (M. munzala) is an endangered and recently discovered cercopithecine primate from Western Arunachal Pradesh, India. On genetic analyses of Arunachal macaques, we observed spatially distributed substantial inter-species genetic divergence among the samples collected from Arunachal Pradesh. The results suggested that Arunachal macaque evolved into two phylogenetic species about 1.96 mya following allopatric speciation by means of Sela mountain pass in Arunachal Pradesh, India. We describe - Sela macaque (M. selai) as a new macaque species that interestingly exhibited high intra-specific genetic variation and also harbors at least two conservation units. Further, we report the past demographic trajectories and quantify genetic variation required for taxonomic clarification. The present study also identifies gap areas for undertaking surveys to document the relic and unknown trans-boundary populations of macaques through multinational, multi-lateral cross border collaboration.

Gene rearrangement in the mitogenome of whiteflies (Hemiptera: Aleyrodinae) along with their phylogeny and characterization of complete mitogenome of Aleurodicus rugioperculatus.

BACKGROUND: Mitochondrial genome rearrangements have been used for defining evolutionary relationships, but there have been incidences of convergences at different taxonomic levels. To gain new insights into whitefly mitogenome evolution, gene arrangement and phylogeny, we sequenced the complete mitogenome of Aleurodicus rugioperculatus (Aleyrodidae: Aleurodicinae) and conducted a comparative analysis with the previously published mitogenomes across the family Aleyrodidae. METHODS AND RESULTS: The complete mitogenome of Aleurodicus rugioperculatus was generated by Next generation sequencing method. It is 15,060 bp circular molecule with 86.5 A + T and 5.5% G + C content. It contains 37 genes (13 PCGs, two rRNAs, 22 tRNAs) and a non-coding control region (CR). Comparative analysis of codon usage indicated that the subfamily Aleyrodinae have weaker bias than Aleurodicinae. Bayesian Inference (BI) and Maximum Likelihood (ML) phylogenetic analyses yielded similar topologies supporting the monophyly of Aleyrodinae and Aleurodicinae. The gene order of 13 whiteflies was compared with ancestor to examine the plesiomorphies, synapomorphies and autapomorphies. We identified five gene blocks (I-V) in the whitefly ancestor that are shared plesiomorphies retained in different whitefly lineages. Gene block I is conserved in all whiteflies except three species. Conversely, we detected 83 derived gene boundaries within the family. Mapping these gene boundaries onto a phylogenetic tree revealed that 16 were symplesiomorphies for two subfamilies; 9 were synapomorphies between the species, and 28 autapomorphies for individual species. CONCLUSIONS: Comparative analyses of gene order of whiteflies revealed the derived gene boundaries which can be further investigated with more mitogenome data to examine the genome evolution in whiteflies.

Structural-genetic insight and optimization of protease production from a novel strain of Aeromonas veronii CMF, a gut isolate of Chrysomya megacephala.

Structural-genetic characterization of protease producing genes and enzymes from microbial sources are seldom appreciated despite having its substantial utilization in protein engineering or genetic manipulation for biotechnological applications. Aeromonas veronii CMF, a mesophilic bacterium isolated from the gut of Chrysomya megacephala, was found to exhibited significant level of protease activity. For the revelation of genetic potential in relation to protease production, whole genome of this organism was sequenced and analysed while structure-function of different protease enzyme was predicated using various in silico analysis. The 4.5 mb CMF genome was found to encompass various types of protease and mostly they are neutral in nature. Enzyme production was highest in an optimum pH and temperature of 6.0 (32.09 ± 1.015 U/ml) and 35ºC (41.65 ± 1.152 U/ml), respectively. Other culture parameters for optimum production of protease were determined to be inoculum size (1%), incubation period (72 h), shaking condition (125 rpm), carbon and nitrogen source [2% lactose (92.21 ± 3.16 U/ml) and 0.5% urea (163.62 ± 4.31 U/ml), respectively] and effect of surfactants [0.02 mg/ml Tween 80 (174.72 ± 4.48 U/ml)]. Furthermore, A. veronii CMF exhibited significant enzyme production like serine protease (15.22 ± 0.563 U/ml), aspartate protease (33.16 ± 0.762 U/ml) and collagenase (17.26 ± 0.626 U/ml). Genomic information and results of physio-biochemical assays indicate its cost-effective potential use in different enzyme-industry.

A new species of Euconocephalus Karny, 1907 (Orthoptera: Tettigoniidae: Conocephalinae: Copiphorini) from India.

A new species, Euconocephalus narayanpurensis Kumar & Chand sp. nov., from India is described in this paper. The new species is similar to the African species Euconocephalus lineatipes (Bolívar, 1890), but differs from the latter in the smaller size, more acute humeral sinus, the narrowly rounded apex of elytra and convex male last abdominal tergite. A key to the Indian species of Euconocephalus Karny, 1907 is also provided.

A new species of the genus Eutyphoeus Michaelsen 1900 (Clitellata: Acanthodrilidae) from the North-Eastern Biogeographic Zone, India.

A new species of the genus Eutyphoeus is described herein from the material collected from Assam, the North-eastern region of India. The new species Eutyphoeus dhubriensis sp. nov. belongs to a group of avestibulate, metandric species, with bidiverticulate spermathecae, lacking lateral intestinal caecae. The proposed species is distinguished from its congeners by the shape, position of genital markings and associated glands, as well as number and position of ventral intestinal caecae and supra-intestinal glands.

A new species of Thinophilus Wahlberg (Diptera: Dolichopodidae: Hydrophorinae) from coastal India with re-description of Thinophilus tesselatus Becker (Diptera: Dolichopodidae: Hydrophorinae).

A new peculiar species of marine dolichopodid fly from West Bengal, India is described: Thinophilus maritimus Mukherjee & Pramanik sp. nov. COI barcodes of the new species were compared to other available Thinophilus sequences. Despite the lack of lateral setae on its hind coxa, this species clusters within the Thinophilus group. Males of Thinophilus tesselatus Becker, 1922 were also obtained from the same area, and a detailed redescription of the species is presented here. A checklist of 70 Oriental Thinophilus species is also provided.

A new species of the genus Hoplochaetella Michaelsen 1900 (Clitellata: Octochaetidae) from the Deccan Peninsula Biogeographic Zone, India.

A new species Hoplochaetella darwini sp. nov. is described from the Deccan Peninsula biogeographic zone of South Indian State, Tamil Nadu. The new species is distinguished from all known Hoplochaetella species by having only one pair of spermathecal pores in segment 7, and a combination of male and prostatic pores in segment 17. The canonical analysis of principal coordinates also supported the proposed new species, showing significant differences with other members of the genus based on the taxonomical characters such as the number and position of spermathecae and prostate glands. Furthermore, an identification key and updated checklist of known species in the genus are also presented along with updated locality details.

Whether curse or blessing: A counterintuitive perspective on global pest thrips infestation under climatic change with implications to agricultural economics.

The improvement and application of pest models to predict yield losses is still a challenge for the scientific community. However, pest models were targeted chiefly towards scheduling scouting or pesticide applications to deal with pest infestation. Thysanoptera (thrips) significantly impact the productivity of many economically important crops worldwide. Until now, no comprehensive study is available on the global distribution of pest thrips, as well as on the extent of cropland vulnerability worldwide. Further, nothing is known about the climate change impacts on these insects. Thus the present study was designed to map the global distribution and quantify the extent of cropland vulnerability in the present and future climate scenarios using data of identified pest thrips within the genus, i.e., Thrips, Frankliniella, and Scirtothrips. Our found significant niche contraction under the climate change scenarios and thrips may reside primarily in their thermal tolerance thresholds. About 3,98,160 km2 of cropland globally was found to be affected in the present scenario. However, it may significantly reduce to 5530 Km2 by 2050 and 1990 km2 by 2070. Further, the thrips distribution mostly getting restricted to Eastern North America, the North-western of the Indian sub-continent, and the north of Europe. Among all realms, thrips may lose ground in the Indo-Malayan realm at the most and get restricted to only 27 out of 825 terrestrial ecoregions. The agrarian communities of the infested regions may get benefit if these pests get wiped out, but on the contrary, we may lose species diversity. Moreover, the vacated niche may attract other invasive species, which may seriously impact the species composition and agricultural productivity. The present study findings can be used in making informed decisions about prioritizing future economic and research investments on the thrips in light of anticipated climate change impacts.

Genetic Characterization and Insular Habitat Enveloping of Endangered Leaf-Nosed Bat, Hipposideros nicobarulae (Mammalia: Chiroptera) in India: Phylogenetic Inference and Conservation Implication.

The Nicobar leaf-nosed Bat (Hipposideros nicobarulae) was described in the early 20th century; however, its systematic classification has been debated for over 100 years. This endangered and endemic species has achieved species status through morphological data in the last 10 years. However, the genetic information and phylogenetic relationships of H. nicobarulae remain neglected. The generated mitochondrial cytochrome b gene (mtCytb) sequences (438 bp) of H. nicobarulae contains 53.42-53.65% AT composition and 1.82% variable sites. The studied species, H. nicobarulae maintains an 8.1% to 22.6% genetic distance from other Hipposideros species. The genetic divergence estimated in this study is congruent with the concept of gene speciation in bats. The Bayesian and Maximum-Likelihood phylogenies clearly discriminated all Hipposideros species and showed a sister relationship between H. nicobarulae and H. cf. antricola. Current mtCytb-based investigations of H. nicobarulae have confirmed the species status at the molecular level. Further, the MaxEnt-based species distribution modelling illustrates the most suitable habitat of H. nicobarulae (294 km2), of which the majority (171 km2) is located on Great Nicobar Island. The present study suggests rigorous sampling across the range, taxonomic coverage, the generation of multiple molecular markers (mitochondrial and nuclear), as well as more ecological information, which will help in understanding population genetic structure, habitat suitability, and the implementation of appropriate conservation action plans for H. nicobarulae and other Hipposideros species.

Morphology and Mitochondrial Lineage Investigations Corroborate the Systematic Status and Pliocene Colonization of Suncus niger (Mammalia: Eulipotyphla) in the Western Ghats Biodiversity Hotspot of India.

The Indian highland shrew, Suncus niger (Horsfield, 1851), is the least studied soricid species from its original range distribution in Southern India, with several systematics conundrums. Following its discovery in 1851, the species was synonymized with Suncus montanus (Kelaart, 1850) (endemic to Sri Lanka) and subsequently identified as a separate Indian population. However, the systematic status of S. niger from topotype specimens in Southern India has yet to be determined through an integrated approach. Both taxonomy and mitochondrial genetic data (Cytochrome b and 16S ribosomal RNA) were used to re-examine the systematics of S. niger. The mtCytb gene clearly distinguished topotypic S. niger from other Suncus species, with high genetic divergences varying from 8.49% to 26.29%. Further, the Bayesian and maximum likelihood topologies clearly segregated S. niger from other congeners and corroborated the sister relationship with S. stoliczkanus with expected divergence in the late Pliocene (2.62 MYA). The TimeTree analysis also exhibits a strong matrilineal affinity of S. dayi (endemic to India) toward the African species. The current study hypothesizes that the ancestor of the soricids evolved in Africa and that genetic lineages were subsequently shifted by plate tectonic events that subsequently colonized different continents as distinct species during the late Miocene (Tortonian) to the Holocene era. In addition to the new range expansion and elevation records of S. niger in the Central Western Ghats, we propose that additional sampling across its distribution, as well as the use of multiple genetic markers, may be useful in determining the genetic diversity and population structure of this endemic species. The present study also recommends that more molecular data on the Soricomorphs lineages, and estimates of their divergence times, will shed light on the evolution of these small mammals on Earth.

African or Asiatic origin? Genome analyses solved the mystery of the hybrid origin of the rescued lion cub.

Species assignment of any seized material using DNA analysis has been a routine and widely accepted standard procedure in providing scientific advisory for the legal prosecution of wildlife cases. Scientific advancements and rigorous application of genetic tools have led to the development of a variety of molecular markers with their defined efficacy in wildlife forensics. However, in a few unusual cases where a hybrid needs to be identified or assignment need to be made at sub-species level, mitochondrial markers often fail or else provide biased results, which can affect the overall judgment in the court of law. Here, we report one such challenging case of lion cub rescued by the law enforcement from illegal trafficking. Phylogenetic assessment based on complete mitogenome assigned rescued lion cub with African lion (Panthera leo leo). However, the TSPY gene of the Y chromosome established that the lion cub shared its paternal lineage from Asiatic lion (Panthera leo persica). With the use of maternally and paternally inherited markers, we conclude a hybrid origin of the rescued lion cub which shared ancestry from both Asiatic as well as African lion. The present study exhibits the application of genome sequencing in thinking beyond routine identification and contributes to the operating procedures of wildlife forensics.

A checklist of black flies (Diptera, Simuliidae) from India.

An updated checklist of the family Simuliidae from India is presented. A total of 79 species of Simulium belonging to eight different subgenera are listed. Eleven species that were not reported in the previous checklist are added here. The present list contributes to a better understanding of the diversity of Simuliidae in India, as well as the impact of Simulium species on the public health of this mega-diverse country.

Prevalence of mouthpart sensilla and protease producing symbiotic gut bacteria in the forensic fly Chrysomya megacephala (Fabricius, 1794): Insight from foraging to digestion.

The blow fly, Chrysomya megacephala (Fabricius, 1794) is a globally prevalent forensically important species that helps to estimate accurate postmortem interval since the death. This fly occasionally causes cutaneous myiasis and transmits several pathogenic bacteria. To understand their ability of corpse detection and digestion of protein-rich meal, the present study describes the mouthpart sensilla and assessment of protease producing symbiotic gut bacteria. Scanning electron microscopy (SEM) showed the prevalence of trichoid sensilla (Tr), basiconic sensilla (Ba) and microtrichia (Mr) on labellar lobes, haustellum and maxillary palps of mouthparts. Bacterial particles of both rod (small and large) and spherical shaped were detected in the gut of C. megacephala using SEM. The bacterial density was higher on the foregut and midgut in comparison to the hindgut. From 72 bacterial isolates, 10 isolates from the foregut region showed considerable protease-producing efficacy ranging between 3.98 - 6.83 GHR and 9.73 - 34.68 U/ml protease. Among these, the most promising protease-producing bacterial isolate showed 16S rDNA sequence similarity (99.85%) with Chryseobacterium artocarpi DNA. This bacterium was the first report from flies. The findings of the study might help in better understanding of the role of sensilla in host perception and foregut symbiotic bacterial association in protein digestion in C. megacephala.

Gut microbial composition in developmental stages of gall inducing thrips Gynaikothrips uzeli and associated plant pathogenesis.

Gut bacteria play a crucial role in the several metabolic activity of the insects. In the present work, effort has been made to decipher the gut microbiota associated with the developmental stages of Gynaikothrips uzeli a gall inducing thrips along with their predicted functional role. Further, an effort has been made to correlate the bacterial communities with plant pathogenesis and thelytoky behaviour of G. uzeli. Findings obtained revealed that genus Arsenophonus dominated the total bacterial diversity and was transmitted vertically through the developmental stages. Further, it was observed that the high abundance of genus Arsenophonus promotes the thelytoky behaviour in G. uzeli and results in the killing of males. Furthermore, strong connecting link between Arsenophonus abundance and gall induction in F. benjamina was observed in the current dataset. G. uzeli being in the category of phloem sucking insect was known for the induction of galls and the current findings for the first time unveiled the facts that high abundance of genus Arsenophonus a well-known plant pathogen may be one of the major reason for inducing galls in F. benjamina. Moreover, PICRUSt2 analysis revealed that predicted functional pathways like biosynthesis of amino acids, and metabolism of carbon, nitrogen, carbohydrates and amino acids (e.g. Arginine, Alanine, Aspartate, Glutamate, Proline, Cysteine, Methionine, Glycine, Threonine, and Serine) were frequently noticed in profiles associated with all the developmental stages of G. uzeli. More to this, the high abundance of Arsenophonus in G. uzeli suggest that representatives of this genus may be resistant and/or tolerant to different antibacterial agents, alkaloids, flavonoids, and glycosides (e.g. quercetin). The correlation of bacterial diversity in pathogenicity can be extrapolated in different pest and vector species of other arthropods.

Complete mitogenome of the endangered and endemic Nicobar treeshrew (Tupaia nicobarica) and comparison with other Scandentians.

The Nicobar treeshrew (Tupaia nicobarica) is an endangered small mammal endemic to the Nicobar Island of the Andaman Sea, India regarded as an alternative experimental animal model in biomedical research. The present study aimed to assemble the first mitochondrial genome of T. nicobarica to elucidate its phylogenetic position with respect to other Scandentians. The structure and variation of the novel mitochondrial genome were analyzed and compared with other Scandentians. The complete mitogenome (17,164 bp) encodes 37 genes, including 13 protein-coding genes (PCGs), 22 transfer RNA (tRNAs), two ribosomal RNA (rRNAs), and one control region (CR). Most of the genes were encoded on majority strand, except nad6 and eight tRNAs. The nonsynonymous/synonymous ratio in all PCGs indicates strong negative selection among all Tupaiidae species. The comparative study of CRs revealed the occurrence of tandem repeats (CGTACA) found in T. nicobarica. The phylogenetic analyses (Maximum Likelihood and Bayesian Inference) showed distinct clustering of T. nicobarica with high branch supports and depict a substantial divergence time (12-19 MYA) from the ancestor lineage of Tupaiidae. The 16S rRNA dataset corroborates the taxonomic rank of two subspecies of T. nicobarica from the Great and Little Nicobar Islands. In the future, whole nuclear genome sequencing is necessary to further improve our understanding of evolutionary relationships among treeshrews, and will have implications for biomedical research.

Life table, survival and fecundity parameters of Aedes albopictus (Diptera: Culicidae) strains from desert and coastal regions of India.

The Asian tiger mosquito, Aedes albopictus (Skuse, 1895) is the anthropophilic and day-biter mosquito, residing in peridomestic habitats and is responsible for vector-borne diseases, i.e., chikungunya and dengue. The knowledge of mosquito life demographics is very important to provide a foundation for a successful vector control program. In the present study, a comparative adult life table, survival, and fecundity parameters were evaluated on desert (Jodhpur, Rajasthan) and coastal (Kolkata, West Bengal) strains of Aedes albopictus (Diptera: Culicidae) from India under the standard laboratory conditions. The results showed significant differences between these strains for the life expectancy of males, fecundity (total eggs/cohort and eggs/female life span), oviposition days, intrinsic rate of increase (rm), finite rate of increase (λ), and doubling time (DT). Female mosquitoes lived longer than the males in both strains. Significant differences were observed in the net reproductive rate (R0) between Kolkata and Jodhpur strains. The coastal (Kolkata) strain showed a longer male life expectancy (5.6 days) than the desert strain (4.0 days). However, the fecundity (eggs/female lifespan) was higher in Kolkata (360.2) than the Jodhpur strain (229.7). The Kolkata strain showed higher R0, rm, λ, and the lower T, DT than the Jodhpur strain. Conclusively, the results indicate that the coastal strain of Ae. albopictus is more r-strategist than the desert strain. This study may help in the precise prediction of Ae. albopictus population dynamics surviving in these geographical areas for vector management.

Nematode community structure along elevation gradient in high altitude vegetation cover of Gangotri National Park (Uttarakhand), India.

The study was undertaken to understand the dynamics of soil nematode communities association along elevation gradient and associated variables in high-altitude regions of Western Himalaya. The diversity and distribution pattern of soil nematodes were investigated at four elevation ranges (Elv1, Elv2, Elv3, and Elv4 each of 500 m class) along altitudinal gradient (3000-5000 m). The nematode community comprised 58 genera of which 37 genera showed sensitivity towards altitudinal variation and the community structure also varied among elevation classes. It was found that elevation significantly affect the nematodes trophic group, diversity composition, and the nematode indices. Nematode generic richness and evenness index indicated a relatively low biodiversity of soil nematodes is supported at higher altitudes. Moreover, maturity indices reflected lower sustenance of k-strategic nematodes at higher elevations. Structure index depicted fewer connections in the soil food web at higher elevations. Nematode faunal profile showed low nutrient and low disturbance in the region. Carbon footprint of the whole nematode assemblage declined along the elevation. Overall substantial differences in the nematode composition, abundance, trophic structure, and contribution to belowground carbon cycling were observed with change in elevation. These findings could be utilized as useful tool in the long-term monitoring and to understand the region's soil health.

Sensory organs of forensically important fly Ophyra capensis (Wiedemann, 1818) (Diptera: Muscidae): A scanning electron microscopic study.

Dipterans, especially the sarcosaprophagous communities are of substantial importance from medical, veterinary and forensic entomological perspectives. Muscids are generally seen to colonize carcasses at advanced stages of decomposition when the initial dominance of calliphorids and sarcophagids subsides. Ophyra capensis (Wiedemann, 1818), a muscid fly with a relatively wide distribution range is considered of decent forensic relevance as it has been reported not only from cadavers placed outdoors but also from graves and exhumed corpses. The prime objective of the present study is to analyse and interpret the ultrastructural morphology of three sensory organs, namely, the ocellar region, compound eye and antenna of adult male and female Ophyra capensis with the help of scanning electron microscopy, so as to facilitate accurate morphological identification of the species in forensic entomological investigations. SEM analysis of the ocellar region revealed that it was larger in size in females and covered with microtrichia. Ultrastructural analysis of the compound eye indicated that the antero-frontally located ommatidia were larger in size in comparison to the rest of the facets, along with notable sexual dimorphism regarding the size of the ommatidia. The ultrastructure of the antenna displayed the presence of five types of sensilla, two types of chaetic sensilla on the scape and pedicel; trichoid sensilla and two types of basiconic sensilla on the flagellum along with numerous microtrichia. Both types of basiconic sensilla displayed a multiporous surface indicating their characteristic olfactory function. The morphological characteristics of these sensilla along with their probable functions are discussed in greater details.