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The wood-feeding termite, Coptotermes formosanus, presents an efficient lignocellulolytic system, offering a distinctive model for the exploration of host-microbial symbiosis towards lignocellulose degradation. Despite decades of investigation, understanding the diversity, community structure, and functional profiles of bacterial symbionts within specific gut regions, particularly the foregut and midgut of C. formosanus, remains largely elusive. In light of this knowledge gap, our efforts focused on elucidating the diversity, community composition and functions of symbiotic bacteria inhabiting the foregut, midgut, and hindgut of C. formosanus via metagenomics. The termite harbored a diverse community of bacterial symbionts encompassing 352 genera and 26 known phyla, exhibiting an uneven distribution across gut regions. Notably, the hindgut displayed a higher relative abundance of phyla such as Bacteroidetes (56.9%) and Spirochetes (23.3%). In contrast, the foregut and midgut were predominantly occupied by Proteobacteria (28.9%) and Firmicutes (21.2%) after Bacteroidetes. The foregut harbored unique phyla like Candidate phylum_TM6 and Armatimonadetes. At the family level, Porphyromonadaceae (28.1, 40.6, and 53.5% abundance in foregut, midgut, and hindgut, respectively) and Spirochaetaceae (foregut = 9%, midgut = 16%, hindgut = 21.6%) emerged as dominant families in the termite's gut regions. Enriched operational taxonomic units (OTUs) were most abundant in the foregut (28), followed by the hindgut (14), while the midgut exhibited enrichment of only two OTUs. Furthermore, the functional analyses revealed distinct influences of bacterial symbionts on various metabolic pathways, particularly carbohydrate and energy metabolisms of the host. Overall, these results underscore significant variations in the structure of the bacterial community among different gut regions of C. formosanus, suggesting unique functional roles of specific bacteria, thereby inspiring further investigations to resolve the crosstalk between host and microbiomes in individual gut-regions of the termite.

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Chironomids are the most abundant aquatic insects in freshwater habitats that can survive in extreme conditions. In this study, as the microbiome provides extended genotype to the host to perform various functions, we explored the microbiota of the Chironomus circumdatus larvae to find out the putative role played by the symbiotic bacteria for the host. The metabarcoding analyses of the larvae revealed that the insect harbors 1771 phylotypes. Out of the various microbial communities found, the majority corresponded to the phyla Proteobacteria (52.59%) and Actinobacteria (20.56%), respectively. The midges also harbored Klebsiella (2.57%), Enterobacter (1.32%), Bacillus (2.29%), and Acinetobacter (2.13%) genera that are involved in detoxification of xenobiotics present in the water. The presence of radiation-resistant genera like Deinococcus, including bacterial species like radiodurans, a highly radiation-resistant bacterium, indicates its potential to support the host's ability to sustain in adverse environments. The functional profiling of the bacteria showed the relative abundance of many enzyme groups, such as transferases (40.62%), oxidoreductases (23.49%), and hydrolases (3.77%). The results indicate that the larvae harbor a considerable variety of bacteria that help the host adapt and survive in the polluted waters. The present study provides thorough insights into the microbiome of the C. circumdatus larvae that can be exploited for the bioremediation of certain pollutants through biomimetic strategies. It also gives us a wake-up call to take a good look at the guts of these disease-carrying insects' inabilities to spread deadly human diseases.

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Phthalates are used as plasticizers in products made of polyvinyl chloride to increase the flexibility of polymers. Unfortunately, these are known to cause adverse effects on living organisms, and also, fast depletion of petroleum resources calls for the exploration of alternatives as replacements. Recent developments in bio-based plasticizers have led to their use as additives for various applications. As they have received much attention in the past decade, it is crucial to study the effects of these plasticizers on living organisms. Hence, we tried to compare the effects of synthetic plasticizer dioctyl phthalate and bio-based plasticizer ethanolamine on Chironomus circumdatus larvae. Mortality was achieved at a lethal concentration (LC50) value of 0.385 mg/L for ethanolamine and dioctyl phthalate at 0.125 mg/L. Disruption in the level of metabolites along with lipid peroxidation was observed in the larvae exposed to plasticizer mediated stress. To overcome these changes, an increase in the levels of antioxidant enzymes such as Superoxide Dismutase, Catalase, Glutathione Peroxidase and Glutathione Reductase, as well as in the levels of detoxifying enzymes like Glutathione-S-Transferase, Esterases and Mixed Function Oxidase during post-exposure recovery conditions was seen. Alterations in the expression levels of heat shock protein 70 and ecdysone receptor genes were also observed. From the comparative study, it could be concluded that Chironomus circumdatus larvae, to a certain extent, have developed tolerance to both ethanolamine and dioctyl phthalate mediated stress. However, dioctyl phthalate has led to more stress as compared to ethanolamine in these larvae.

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Insects harbor bacteria that are a rich source of enzymes that can be utilized for various industrial and biotechnological applications. It is predicted that during evolution these detoxifying enzymes have been acquired by insects through bacteria. However, the role of host insect detoxification enzymes has already been extensively studied but only a few resistance development studies have been focused on the enzymes derived from gut bacteria. Thus in this study, two bacterial isolates were found in the gut of Chironomous circumdatus larvae having esterase activity, out of which one bacterium was molecular characterized and it was found to be Enterobacter mori designated as strain BI1245. Further, crude extract from the bacterium was characterized and it was observed that it showed maximum activity at pH 8 and temperature 60 °C. Moreover, the crude extract showed 0.26 mM and 290 U/mg of protein as Km and Vmax value when p-nitrophenyl acetate was used as a substrate. Thereafter, the bacterial isolate was incubated in minimal salts medium containing modified phthalates and it was found that bacterium could utilize shorter alkyl-chain phthalic acid esters faster as compared to long alkyl chains thereby indicating that side chain of the substrates has a significant effect on the utilization of phthalic acid esters. Thus the gut flora present in insects may play an important role in providing resistance to the host to live in phthalate polluted water.

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Plasticizers are used as additives in making plastics. Diethyl phthalate (DEP) is one of the majorly used plasticizers in various products. When plastic materials are dumped in an aquatic system, there is an increase in chances of DEP leaching out and getting deposited in water. Thus the current work focuses on studying the effect of DEP on the larval stages of Chironomus circumdatus. In this study it was found that there was an increase in lipid peroxidation levels indicating imposition of oxidative stress on these larvae due to the exposure of DEP. Changes in the levels of carbohydrates and lipids were also seen. To reduce these effects antioxidant defense system may get activated. Thus investigations showed an increase in enzymatic activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and acetylcholinesterases (AchE) and decrease in the activity of glutathione reductase (GR). Nonenzymatic antioxidant glutathione levels were also increased during the post-recovery exposure period. Thus this indicates that both enzymatic, as well as non-enzymatic antioxidants, play a certain role in reducing the stress mediated by DEP. Up-regulation of gene expression of heat shock protein70 (hsp70) was observed, which is one of the conserved protein produced during stress response in many dipterans. Changes in the level of expression of the ecdysone receptor (EcR) gene were also seen in DEP exposed larvae. Thus the insights give us a preliminary indication that physiological and developmental adaptations may take place in these organisms to persist in the DEP contaminated environment. CAPSULE: DEP mediated stress imposes changes in the metabolites and thus activation of antioxidant defense system in aquatic midges of Chironomus circumdatus. Changes in the expression of heat shock protein70 and ecdysone receptor was also seen indicating that DEP mediated stress affects at the molecular level also of the organism. These changes may help them to tolerate and live in DEP polluted water.

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