Elucidating the structure, and composition of bacterial symbionts in the gut regions of wood-feeding termite, Coptotermes formosanus and their functional profile towards lignocellulolytic systems.

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.

Host-Specific Diversity of Culturable Bacteria in the Gut Systems of Fungus-Growing Termites and Their Potential Functions towards Lignocellulose Bioconversion.

Fungus-growing termites are eusocial insects that represent one of the most efficient and unique systems for lignocellulose bioconversion, evolved from a sophisticated symbiosis with lignocellulolytic fungi and gut bacterial communities. Despite a plethora of information generated during the last century, some essential information on gut bacterial profiles and their unique contributions to wood digestion in some fungus-growing termites is still inadequate. Hence, using the culture-dependent approach, the present study aims to assess and compare the diversity of lignocellulose-degrading bacterial symbionts within the gut systems of three fungus-growing termites: Ancistrotermes pakistanicus, Odontotermes longignathus, and Macrotermes sp. A total of 32 bacterial species, belonging to 18 genera and 10 different families, were successfully isolated and identified from three fungus-growing termites using Avicel or xylan as the sole source of carbon. Enterobacteriaceae was the most dominant family represented by 68.1% of the total bacteria, followed by Yersiniaceae (10.6%) and Moraxellaceae (9%). Interestingly, five bacterial genera such as Enterobacter, Citrobacter, Acinetobacter, Trabulsiella, and Kluyvera were common among the tested termites, while the other bacteria demonstrated a termite-specific distribution. Further, the lignocellulolytic potential of selected bacterial strains was tested on agricultural waste to evaluate their capability for lignocellulose bioconversion. The highest substrate degradation was achieved with E. chengduensis MA11 which degraded 45.52% of rice straw. All of the potential strains showed endoglucanase, exoglucanase, and xylanase activities depicting a symbiotic role towards the lignocellulose digestion within the termite gut. The above results indicated that fungus-growing termites harbor a diverse array of bacterial symbionts that differ from species to species, which may play an inevitable role to enhance the degradation efficacy in lignocellulose decomposition. The present study further elaborates our knowledge about the termite-bacteria symbiosis for lignocellulose bioconversion which could be helpful to design a future biorefinery.

Mining the diversity and functional profile of bacterial symbionts from the larvae of Chironomus circumdatus (bloodworms).

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.

Exploring the region-wise diversity and functions of symbiotic bacteria in the gut system of wood-feeding termite, Coptotermes formosanus, toward the degradation of cellulose, hemicellulose, and organic dyes.

The wood-feeding termite Coptotermes formosanus represents a unique and impressive system for lignocellulose degradation. The highly efficient digestion of lignocellulose is achieved through symbiosis with gut symbionts like bacteria. Despite extensive research during the last three decades, diversity of bacterial symbionts residing in individual gut regions of the termite and their associated functions is still lacking. To this end, cellulose, xylan, and dye-decolorization bacteria residing in foregut, midgut, and hindgut regions of C. formosanus were enlisted by using enrichment and culture-dependent molecular methods. A total of 87 bacterial strains were successfully isolated from different gut regions of C. formosanus which belonged to 27 different species of 10 genera, majorly affiliated with Proteobacteria (80%) and Firmicutes (18.3%). Among the gut regions, 37.9% of the total bacterial isolates were observed in the hindgut that demonstrated predominance of cellulolytic bacteria (47.6%). The majority of the xylanolytic and dye-decolorization bacteria (50%) were obtained from the foregut and midgut, respectively. Actinobacteria represented by Dietza sp. was observed in the hindgut only. Based on species richness, the highest diversity was observed in midgut and hindgut regions each of which harbored seven unique bacterial species. The members of Enterobacter, Klebsiella, and Pseudomonas were common among the gut regions. The lignocellulolytic activities of the selected potential bacteria signpost their assistance to the host for lignocellulose digestion. The overall results indicate that C. formosanus harbors diverse communities of lignocellulolytic bacteria in different regions of the gut system. These observations will significantly advance our understanding of the termite-bacteria symbiosis and their microbial ecology uniquely existed in different gut regions of C. formosanus, which may further shed a light on its potential values at termite-modeled biotechnology.

Bio-based versus synthetic: comparative study of plasticizers mediated stress on Chironomus circumdatus (Diptera-Chironomidae).

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.

Valorization Potential of a Novel Bacterial Strain, Bacillus altitudinis RSP75, towards Lignocellulose Bioconversion: An Assessment of Symbiotic Bacteria from the Stored Grain Pest, Tribolium castaneum.

Bioconversion of lignocellulose into renewable energy and commodity products faces a major obstacle of inefficient saccharification due to its recalcitrant structure. In nature, lignocellulose is efficiently degraded by some insects, including termites and beetles, potentially due to the contribution from symbiotic gut bacteria. To this end, the presented investigation reports the isolation and characterization of cellulolytic bacteria from the gut system of red flour beetle, Tribolium castaneum. Out of the 15 isolated bacteria, strain RSP75 showed the highest cellulolytic activities by forming a clearance zone of 28 mm in diameter with a hydrolytic capacity of ~4.7. The MALDI-TOF biotyping and 16S rRNA gene sequencing revealed that the strain RSP75 belongs to Bacillus altitudinis. Among the tested enzymes, B. altitudinis RSP75 showed maximum activity of 63.2 IU/mL extract for xylanase followed by ß-glucosidase (47.1 ± 3 IU/mL extract) which were manifold higher than previously reported activities. The highest substrate degradation was achieved with wheat husk and corn cob powder which accounted for 69.2% and 54.5%, respectively. The scanning electron microscopy showed adhesion of the bacterial cells with the substrate which was further substantiated by FTIR analysis that depicted the absence of the characteristic cellulose bands at wave numbers 1247, 1375, and 1735 cm-1 due to hydrolysis by the bacterium. Furthermore, B. altitudinis RSP75 showed co-culturing competence with Saccharomyces cerevisiae for bioethanol production from lignocellulose as revealed by GC-MS analysis. The overall observations signify the gut of T. castaneum as a unique and impressive reservoir to prospect for lignocellulose-degrading bacteria that can have many biotechnological applications, including biofuels and biorefinery.

Characterization of an Esterase Producing Bacterium from the Gut of Chironomus circumdatus (Bloodworms) and its Ability to Use Modified Phthalates.

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.

Recent advances in the bioprospection and applications of chitinolytic bacteria for valorization of waste chitin.

One of the most abundant natural polymers on earth, chitin is a fibrous and structural polysaccharide, composed of N-acetyl-D-glucosamine. The biopolymer is the major structural constituent of fungi, arthropods, mollusks, nematodes, and some algae. The biodegradation of chitin is largely manifested by chitinolytic enzyme secreting organisms including bacteria, insects, and plants. Among them, bacterial chitinases represent the most promising, inexpensive, and sustainable source of proteins that can be employed for industrial-scale applications. To this end, the presented review comes at a timely moment to highlight the major sources of chitinolytic bacteria. It also discusses the potential pros and cons of prospecting bacterial chitinases that can be easily manipulated through genetic engineering. Additionally, we have elaborated the recent applications of the chitin thereby branding chitinases as potential candidates for biorefinery and biomedical research for eco-friendly and sustainable management of chitin waste in the environment.

Bioefficacy of Lemongrass and Tea Tree Essential Oils Against House Fly, Musca domestica.

ABSTRACT: The management of house fly, Musca domestica has become immensely important to prevent epidemics of many detrimental diseases. In view of this objective, the present study demonstrates the efficacy of lemongrass (LG) and tea tree essential oils (TTEOs) against M. domestica. The TTEO proved to be more lethal against larvae and adults of M. domestica depicting an LC50 at 14.88 mg/ dm3 which was 17.19 mg/ dm3 for LGEO. In contrast, pupicidal effect of LGEO was much higher (LD50, 14.49 µl/0.25L) as compared to TTEO. The LGEO drastically reduced the total body sugar, glycogen and protein contents by 3.29, 2.95 and 7.56 fold, respectively, contrasting with high influence of TTEO on lipid content of the late 3rd instar larvae. A considerable reduction in gut enzymes secretion was observed due to treatment of EOs thereby altering gut physiology of the insect. Moreover, significant inhibition of acetylcholine esterase (AchE) was also observed with LGEO at LC50 concentration (5.33 mg/ml) inhibiting insect neurotransmission. The gas chromatography-mass spectrometric (GC-MS) analysis of the LGEO showed 12 major compounds dominated by Citral whereas TTEO contained only 5 major compounds. Further analysis by field emission scanning electron microscopy (FESEM) revealed distortion and shrinkage of larval bodies caused by the treatment of EOs. These overall observations brand LG and TT-EOs as potential organic-insecticides against M. domestica. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s40011-020-01220-z) contains supplementary material, which is available to authorized users.

Biochemical and molecular changes mediated by plasticizer diethyl phthalate in Chironomus circumdatus (bloodworms).

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.