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Sixteen isolates of bacteria obtained from the entomopathogenic nematode (Heterorhabditis sp.) infected cadavers of Galleria mellonella larvae were identified following phenotypic characterization and molecular analysis of 16S rRNA. Two isolates were identified as the symbiotic bacterium, Photothabdus luminescens, while 14 other isolates were represented by nine species of nonsymbiotic bacteria viz., Stenotrophomonas maltophilia, Alcaligenes aquatilis, Brevundimonas diminuta, Brucella pseudointermedia, Ochrobactrum sp., Brucella pseudogrignonensis, Brucella anthropic, Pseudomonas azatoformans and Pseudomonas lactis. The phylogenetic analysis confirmed the evolutionary relationship between P. luminescens and Pseudomonas spp. The study also found a close relationship among the nonsymbiotic bacteria such as A. aquatilis, B. diminuta, Ochrobactrum sp., and Brucella spp. P. luminescens has been documented for its insecticidal effects against a wide range of insects. The two local isolates obtained in this study may be explored for their biocontrol potential against major pests of the region. Further, the association of nonsymbiotic bacteria with the EPN may be investigated. [ABSTRACT FROM AUTHOR]

Symbiotic gut bacteria play crucial role in host health. Symbionts are widely distributed in arthropod intestines, but their ecological functions are poorly understood due to the inability to cultivate them. Members of Candidatus Bacilliplasma (CB) are widely distributed in crustacean intestine and maybe commensals with hosts, but the paucity of pure cultures has limited further insights into their physiologies and functions. Here, four strains of representative CB bacteria in shrimp intestine were successfully isolated and identified as members of a novel Order in the Phylum Mycoplasmatota. Through genome assembly, the circular genome maps of the four strains were obtained, and the number of coding genes ranged from 1,886 to 1,980. Genomic analysis suggested that the bacteria were missing genes for many critical pathways including the TCA cycle and biosynthesis pathways for amino acids and coenzyme factors. The analysis of 16S amplification data showed that Shewanella, Pseudomonas and CB were the dominant at the genera level in the intestine of Penaeus vannamei. Ecological functional experiments revealed that the strains were symbionts and colonized shrimp intestines. Our valued findings can greatly enhance our understanding and provides new insights into the potentially significant role of uncultured symbiotic bacteria in modulating host health. [ABSTRACT FROM AUTHOR]

Background: Bacteria of the candidate phyla radiation (CPR), constituting about 25% of the bacterial biodiversity, are characterized by small cell size and patchy genomes without complete key metabolic pathways, suggesting a symbiotic lifestyle. Gracilibacteria (BD1-5), which are part of the CPR branch, possess alternate coded genomes and have not yet been cultivated. The lifestyle of Gracilibacteria, their temporal dynamics, and activity in natural ecosystems, particularly in groundwater, has remained largely unexplored. Here, we aimed to investigate Gracilibacteria activity in situ and to discern their lifestyle based on expressed genes, using the metaproteogenome of Gracilibacteria as a function of time in the cold-water geyser Wallender Born in the Volcanic Eifel region in Germany. Results: We coupled genome-resolved metagenomics and metaproteomics to investigate a cold-water geyser microbial community enriched in Gracilibacteria across a 12-day time-series. Groundwater was collected and sequentially filtered to fraction CPR and other bacteria. Based on 725 Gbps of metagenomic data, 1129 different ribosomal protein S3 marker genes, and 751 high-quality genomes (123 population genomes after dereplication), we identified dominant bacteria belonging to Gallionellales and Gracilibacteria along with keystone microbes, which were low in genomic abundance but substantially contributing to proteomic abundance. Seven high-quality Gracilibacteria genomes showed typical limitations, such as limited amino acid or nucleotide synthesis, in their central metabolism but no co-occurrence with potential hosts. The genomes of these Gracilibacteria were encoded for a high number of proteins involved in cell to cell interaction, supporting the previously surmised host-dependent lifestyle, e.g., type IV and type II secretion system subunits, transporters, and features related to cell motility, which were also detected on protein level. Conclusions: We here identified microbial keystone taxa in a high-CO2 aquifer, and revealed microbial dynamics of Gracilibacteria. Although Gracilibacteria in this ecosystem did not appear to target specific organisms in this ecosystem due to lack of co-occurrence despite enrichment on 0.2-µm filter fraction, we provide proteomic evidence for the complex machinery behind the host-dependent lifestyle of groundwater Gracilibacteria. 38kgAxaMce3Yn9ghud_S2p Video Abstract [ABSTRACT FROM AUTHOR]

Many insects are obligatorily associated with and dependent on specific microbial species as essential mutualistic partners. In the host insects, such microbial mutualists are usually maintained in specialized cells or organs, called bacteriocytes or symbiotic organs. Hence, potentially exponential microbial growth cannot be realized but must be strongly constrained by spatial and resource limitations within the host cells or tissues. How such endosymbiotic bacteria grow, divide, and proliferate is important for understanding the interactions and dynamics underpinning intimate host-microbe symbiotic associations. Here we report that Blattabacterium, the ancient and essential endosymbiont of cockroaches, exhibits unexpectedly high rates of cell division (20%-58%) and, in addition, the cell division is asymmetric (average asymmetry index >1.5) when isolated from the German cockroach Blattella germanica. The asymmetric division of endosymbiont cells at high frequencies was observed irrespective of host tissues (fat bodies vs ovaries) or developmental stages (adults vs nymphs vs embryos) of B. germanica, and also observed in several different cockroach species. By contrast, such asymmetric and frequent cell division was observed neither in Buchnera, the obligatory bacterial endosymbiont of aphids, nor in Pantoea, the obligatory bacterial gut symbiont of stinkbugs. Comparative genomics of cell division-related genes uncovered that the Blattabacterium genome lacks the Min system genes that determine the cell division plane, which may be relevant to asymmetric cell division. These observations combined with comparative symbiont genomics provide insight into what processes and regulations may underpin the growth, division, and proliferation of such bacterial mutualists continuously constrained under within-host conditions. IMPORTANCE Diverse insects are dependent on specific bacterial mutualists for their survival and reproduction. Due to the long-lasting coevolutionary history, such symbiotic bacteria tend to exhibit degenerative genomes and suffer uncultivability. Because of their microbiological fastidiousness, the cell division patterns of such uncultivable symbiotic bacteria have been poorly described. Here, using fine microscopic and quantitative morphometric approaches, we report that, although bacterial cell division usually proceeds through symmetric binary fission, Blattabacterium, the ancient and essential endosymbiont of cockroaches, exhibits frequent and asymmetric cell division. Such peculiar cell division patterns were not observed with other uncultivable essential symbiotic bacteria of aphids and stinkbugs. Gene repertoire analysis revealed that the molecular machinery for regulating the bacterial cell division plane are lost in the Blattabacterium genome, suggesting the possibility that the general trend toward the reductive genome evolution of symbiotic bacteria may underpin their bizarre cytological/morphological traits. [ABSTRACT FROM AUTHOR]

Microorganisms colonize all barrier tissues and are present on the skin and all mucous membranes from birth. Bacteria have many ways of influencing the host organism, including activation of innate immunity receptors by pathogen-associated molecular patterns and synthesis of various chemical compounds, such as vitamins, short-chain fatty acids, bacteriocins, toxins. Bacteria, using extracellular vesicles, can also introduce high-molecular compounds, such as proteins and nucleic acids, into the cell, regulating the metabolic pathways of the host cells. Epithelial cells and immune cells recognize bacterial bioregulators and, depending on the microenvironment and context, determine the direction and intensity of the immune response. A large number of factors influence the maintenance of symbiotic microflora, the diversity of which protects hosts against pathogen colonization. Reduced bacterial diversity is associated with pathogen dominance and allergic diseases of the skin, gastrointestinal tract, and upper and lower respiratory tract, as seen in atopic dermatitis, allergic rhinitis, chronic rhinosinusitis, food allergies, and asthma. Understanding the multifactorial influence of microflora on maintaining health and disease determines the effectiveness of therapy and disease prevention and changes our food preferences and lifestyle to maintain health and active longevity. [ABSTRACT FROM AUTHOR]

ABSTRACT Aphids are serious insect pests for agricultural and horticultural crops and may cause the major economic losses. Insecticides used to control aphids have caused environmental pollution and the insecticide residues in agricultural products. A new protease inhibitor gene from bacterium Xenorhabdus bovienii (Xbpi‐1) and the protease inhibitor protein expressed by the gene against the pea aphid (Acyrthosiphon pisum Harris; Hemiptera: Aphididae) have been reported, however, effects of the PIP on the symbiotic bacteria, Buchnera aphidicola, and gene expression in pea aphids are unknown. By assessing the quantity of B. aphidicola, the primary symbiotic bacterium, in the aphid fed on an artificial diet containing 100 and 500 μg/mL Xbpi‐1, we observed a substantial reduction in its population by 27% and 46%, respectively, as analysed by real‐time quantitative reverse transcription PCR (qRT‐PCR). Furthermore, the growth of other aphid‐associated bacteria was also significantly inhibited by Xbpi‐1. To elucidate the mechanisms at the gene level, we conducted transcriptome analysis and identified differentially expressed genes (DEGs). Subsequent Gene Ontology (GO) analysis of the 213 DEGs shed light on the impact of Xbpi‐1 on aphid metabolism processes and gene expression. Notably, the results highlighted several aphid nutrient metabolism pathways affected by Xbpi‐1, which are relevant to vector‐borne diseases. These pathways encompass crucial factors such as heat shock proteins, cuticle proteins and proteases. The results from this study revealed that the PIP had a novel mechanism against pea aphids by having adverse effects on the primary symbiotic bacteria in pea aphids and affecting aphid gene expression, showing that the PIP may be a promise bioinsecticide for aphid control in the future. [ABSTRACT FROM AUTHOR]

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Abstract Background Bacteria of the candidate phyla radiation (CPR), constituting about 25% of the bacterial biodiversity, are characterized by small cell size and patchy genomes without complete key metabolic pathways, suggesting a symbiotic lifestyle. Gracilibacteria (BD1-5), which are part of the CPR branch, possess alternate coded genomes and have not yet been cultivated. The lifestyle of Gracilibacteria, their temporal dynamics, and activity in natural ecosystems, particularly in groundwater, has remained largely unexplored. Here, we aimed to investigate Gracilibacteria activity in situ and to discern their lifestyle based on expressed genes, using the metaproteogenome of Gracilibacteria as a function of time in the cold-water geyser Wallender Born in the Volcanic Eifel region in Germany. Results We coupled genome-resolved metagenomics and metaproteomics to investigate a cold-water geyser microbial community enriched in Gracilibacteria across a 12-day time-series. Groundwater was collected and sequentially filtered to fraction CPR and other bacteria. Based on 725 Gbps of metagenomic data, 1129 different ribosomal protein S3 marker genes, and 751 high-quality genomes (123 population genomes after dereplication), we identified dominant bacteria belonging to Gallionellales and Gracilibacteria along with keystone microbes, which were low in genomic abundance but substantially contributing to proteomic abundance. Seven high-quality Gracilibacteria genomes showed typical limitations, such as limited amino acid or nucleotide synthesis, in their central metabolism but no co-occurrence with potential hosts. The genomes of these Gracilibacteria were encoded for a high number of proteins involved in cell to cell interaction, supporting the previously surmised host-dependent lifestyle, e.g., type IV and type II secretion system subunits, transporters, and features related to cell motility, which were also detected on protein level. Conclusions We here identified microbial keystone taxa in a high-CO2 aquifer, and revealed microbial dynamics of Gracilibacteria. Although Gracilibacteria in this ecosystem did not appear to target specific organisms in this ecosystem due to lack of co-occurrence despite enrichment on 0.2-µm filter fraction, we provide proteomic evidence for the complex machinery behind the host-dependent lifestyle of groundwater Gracilibacteria. Video Abstract

Biological nitrogen fixation (BNF) can provide the necessary nitrogen for bean crops; however, for this to occur, important limitations involving the inoculant application technology need to be overcome.The use of co-inoculation is a management technique used to obtain benefits and increase the potential of N2 fixation from the association between bacteria from the rhizobia group, such as R. tropici, and bacteria that promote plant growth, such as A. brasilense, in association with the addition of nutrients that allow greater efficiency of bacteria fixing atmospheric N2. This study aimed to evaluate the bean response to the reinoculation of R. tropici in co-inoculation with A. brasilense in a mixture with the micronutrients Co/Mo, in the winter season of 2021, in Anápolis-GO, Brazil. A randomized block design was used, with four replications, and the following treatments (TRs) were studied: TR1—reinoculation with R. tropici; TR2—reinoculation with co-inoculation of R. tropici + A. brasilense; TR3—reinoculation of R. tropici + Mo/Co micronutrients; TR4—reinoculation with co-inoculation R. tropici + A. brasilense + Mo/Co micronutrients; TR5—inoculation via seed, without reinoculation; TR6—mineral N fertilization in the sowing furrow and topdressing; TR7—control, without any N source. At stage R6, nodulation characteristics (number and dry mass of nodules) and the morphophysiological parameters of the plants (main root length, root dry mass, plant height, shoot dry mass, leaf area, and leaf N content in the shoot) were evaluated. At harvest, the final plant stand and components (number of pods per plant, number of grains per pod, and average weight of one hundred grains) were determined, in addition to grain yield. It was concluded that inoculation followed by reinoculation in topdressing with R. tropici in co-inoculation with A. brasilense plus Mo/Co, compared to mineral nitrogen fertilization, improves the efficiency of the nodulation process and the morphophysiological characteristics of the common bean crop. Seed inoculation and topdressing application with R. tropici, associated with co-inoculation with A. brasilense + Mo and Co, have the potential to completely replace mineral nitrogen fertilization in common bean crops. [ABSTRACT FROM AUTHOR]

The common cutworm, Spodoptera litura, is a serious insect pest of many vegetables and crops worldwide. Entomopathogenic nematodes (EPN) have been utilized as biological control agents for controlling various insect pests, including the larvae of S. litura. Many indigenous EPN have been recognized to be more effective in specific field applications. Among the 160 soil samples collected in undisturbed areas of western Thailand, three samples tested positive for EPN. Three indigenous EPN were identified as Steinernema siamkayai namely, EPNKU63, EPNKU70 and EPNKU85, based on ITS and D2/D3 expansion region analysis of the 28s rRNA genes. Additionally, genetic analysis of the symbiotic bacteria using recA rRNA sequences confirmed their identity as Xenorhabdus stockiae namely, PEPNKU63, PEPNKU70 and PEPNKU85. To evaluate their initial biocontrol potential against the 6th instar larvae of Galleria mellonella, virulence assays were conducted. The application of 100 infective juveniles (IJs)/insects resulted in the mortality of 80–100% of G. mellonella larvae after 72 h. When symbiotic bacteria were applied at 1 × 106 cells/insect, they exhibited 63–93% mortality against G. mellonella larvae after 120 h. In further laboratory tests, three S. siamkayai isolates achieved 100% mortality of 3rd instar Spodoptera litura larvae within 72 h, with LC50 values ranging from 29 to 30 IJs/insect. In screenhouse experiments, it was revealed that all S. siamkayai isolates displayed substantial virulence, ranging from 62% to 74%, against 3rd instar S. litura larvae within 72 h. This study demonstrates the biocontrol potential of S. siamkayai in controlling S. litura larvae. [ABSTRACT FROM AUTHOR]

BACKGROUND: The potential of copper nanoparticles (Cu‐NPs) to be used as an alternative control strategy against olive fruit flies (Bactrocera oleae) with reduced sensitivity to the pyrethroid deltamethrin and the impact of both nanosized and bulk copper hydroxide (Cu(OH)2) on the insect's reproductive and endosymbiotic parameters were investigated. RESULTS: The application of nanosized and bulk copper applied by feeding resulted in significant levels of adult mortality, comparable to or surpassing those achieved with deltamethrin at recommended doses. Combinations of Cu‐NPs or copper oxide nanoparticles (CuO‐NPs) with deltamethrin significantly enhanced the insecticide's efficacy against B. oleae adults. When combined with deltamethrin, Cu‐NPs significantly reduced the mean total number of offspring compared with the control, and the number of stings, pupae, female and total number of offspring compared with the insecticide alone. Both bulk and nanosized copper negatively affected the abundance of the endosymbiotic bacterium Candidatus Erwinia dacicola which is crucial for the survival of B. oleae larvae. CONCLUSION: The Cu‐NPs can aid the control of B. oleae both by reducing larval survival and by enhancing deltamethrin performance in terms of toxicity and reduced fecundity, providing an effective anti‐resistance tool and minimizing the environmental footprint of synthetic pesticides by reducing the required doses for the control of the pest. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Seed inoculation with symbiotic bacteria is a commonly employed practice in soybean cultivation. As a result, nodulation proceeds properly and plants self-supply atmospheric nitrogen, requiring either minimal or no additional nitrogen fertilization. The aim of the study was to investigate the response of soybeans to the application of the recommended or double dose of commercial inoculants (HiStick® Soy or TURBOSOY®) and/or mineral nitrogen fertilization compared to the untreated control. It was demonstrated that a double dose of the tested preparations had the most favorable effect on nodulation. However, the impact of weather conditions modified their effectiveness during the study years, which was especially visible in 2022. Sowing seeds without inoculation (control) resulted in the formation of sparse root nodules and consequently the lowest leaf area index (LAI) and soil plant analysis development (SPAD) measurements. In addition, the values of SPAD and LAI indices varied across the years of the study, indicating that weather conditions modified nitrogen uptake by plants. Overall, seed inoculation and/or nitrogen fertilization positively influenced the chemical composition of seeds compared to the control. The only decrease observed was in the oil content, while the double dose of HiStick® Soy preparation reduced the polyphenol content. The double dose of the tested inoculants had the most favorable impact on yield components and seed yield. However, applying inoculation at the recommended dose or in combination with nitrogen fertilization yielded similar or slightly worse results, depending on the year. Therefore, soybean seed inoculation should be recommended, although the effectiveness of the procedure will depend on various factors, including the type of inoculant, dosage, nitrogen fertilization, and weather conditions. [ABSTRACT FROM AUTHOR]

Secondary metabolites, bioactive compounds produced by living organisms, can unveil symbiotic relationships in nature. In this study, soilborne entomopathogenic nematodes associated with symbiotic bacteria (Xenorhabdus stockiae and Photorhabdus luminescens) were extracted from solvent supernatant containing secondary metabolites, demonstrating significant inhibitory effects against E. coli, S. aureus, B. subtilus, P. mirabilis, E. faecalis, and P. stutzeri. The characterization of these secondary metabolites by Fourier transforms infrared spectroscopy revealed amine groups of proteins, hydroxyl and carboxyl groups of polyphenols, hydroxyl groups of polysaccharides, and carboxyl groups of organic acids. Furthermore, the obtained crude extracts were analyzed by high-performance liquid chromatography for the basic identification of potential bioactive peptides. Gas chromatography–mass spectrometry analysis of ethyl acetate extracts from Xenorhabdus stockiae identified major compounds including nonanoic acid derivatives, proline, paromycin, octodecanal derivatives, trioxa-5-aza-1-silabicyclo, 4-octadecenal, methyl ester, oleic acid, and 1,2-benzenedicarboxylicacid. Additional extraction from Photorhabdus luminescens yielded functional compounds such as indole-3-acetic acid, phthalic acid, 1-tetradecanol, nemorosonol, 1-eicosanol, and unsaturated fatty acids. These findings support the potential development of novel natural antimicrobial agents for future pathogen suppression. [ABSTRACT FROM AUTHOR]

Abstract Background Karst caves serve as natural laboratories, providing organisms with extreme and constant conditions that promote isolation, resulting in a genetic relationship and living environment that is significantly different from those outside the cave. However, research on cave creatures, especially Opiliones, remains scarce, with most studies focused on water, soil, and cave sediments. Results The structure of symbiotic bacteria in different caves were compared, revealing significant differences. Based on the alpha and beta diversity, symbiotic bacteria abundance and diversity in the cave were similar, but the structure of symbiotic bacteria differed inside and outside the cave. Microorganisms in the cave play an important role in material cycling and energy flow, particularly in the nitrogen cycle. Although microbial diversity varies inside and outside the cave, Opiliones in Beijing caves and Hainan Island exhibited a strong similarity, indicating that the two environments share commonalities. Conclusions The karst cave environment possesses high microbial diversity and there are noticeable differences among different caves. Different habitats lead to significant differences in the symbiotic bacteria in Opiliones inside and outside the cave, and cave microorganisms have made efforts to adapt to extreme environments. The similarity in symbiotic bacteria community structure suggests a potential similarity in host environments, providing an explanation for the appearance of Sinonychia martensi in caves in the north.

Insecticides play a crucial role in safeguarding crops from pest. However, prior to their release, it is essential to assess the efficacy and potential effects of these protectants. Flupyradifurone (FBF) and sulfoxaflor (SFX), two insecticides commonly used to control aphids, have been found to exert negative effects on the growth and development of Binodoxys communis Gahan, a beneficial parasitic wasp that targets aphids. However, it remains unknown if FBF and SFX influence the symbiotic bacteria residing within B. communis. In this study, 16SrRNA sequencing was utilized to assess the populations of B. communis symbionts exposed to sublethal concentrations (LC10 and LC25) of FBF and SFX. The control and treatment groups exhibited similar bacterial community structures, with Bacteroidota identified as the dominant phylum in each. After exposure to FBF and SFX, the diversity and richness of symbionts (Firmicutes, Bacteroidota, and Actinobacteriota) in B. communis changed notably. The two different insecticides and exposure durations exerted distinct effects on the microbial community. For instance, SFX stress resulted in a decrease in Akkermansia and an increase in Escherichia Shigella 1h after exposure, with both returning to baseline levels after 3d. In contrast, FBF exposure caused a reduction in Akkermansia presence after 1h, which persisted even after 3d. This study represents the first assessment of the effects of FBF and SFX exposure on symbiotic bacteria in B. communis, expanding our understanding of how insecticides influence natural enemies and their symbiotic bacterial relationships. This study provides theoretical guidance for field applications of FBF and SFX, as well as a reasonable basis for exploring chemical resistance conferred by symbiotic bacteria.

Green macroalgae, known locally as Latoh, is one of the green seaweeds consumed by the local community in Jepara and is beneficial for health. This study explores the potential of secondary metabolites from seaweed and its symbiotic bacteria as natural food preservatives and antibacterials. Seaweed samples were collected from the seagrass ecosystem of Panjang Island, Jepara, Indonesia. Subsequently, the samples were subjected to scanning electron microscopy analysis, proximate analysis, phytochemical analysis, thin layer chromatography, and high-performance liquid chromatography for amino acid analysis. A sample was subjected to a multistage extraction process using n-hexane, ethyl acetate, and methanol (1:5 w/v), each for 24 h. Symbiotic bacteria from seaweed were isolated, and enzymatic (proteolytic, amylolytic, and cellulolytic) and antibacterial testing against pathogenic bacteria Staphylococcus aureus and Escherichia coli was conducted using the disc diffusion method. The selected bacteria were subjected to molecular identification. The research showed that Caulerpa lentillifera had an ash content of 3.24 %, protein content of 0.57 %, and fat content of 0.337 %. Phytochemical analysis shows that the sample contains flavonoids, steroids, and alkaloids. HPLC analysis reveals that Caulerpa lentillifera has the highest content of aspartic acid (relative area: 11.90 %), glutamic acid (relative area: 13.43 %), and alanine (relative area: 9.03 %). Caulerpa racemosa sample shows the highest detector response for glutamic acid (relative area: 12.19%), aspartic acid (relative area: 11.10 %), and alanine (relative area: 9.63 %). The results indicate that 14 bacterial isolates were successfully isolated, with 6 isolates from Caulerpa lentillifera and 8 isolates from Caulerpa racemosa, all exhibiting enzymatic and antibacterial abilities. The research results concluded that the Latoh seaweed species Caulerpa lentillifera and Caulerpa racemosa and their symbiotic bacteria have the potential to be used as food preservatives. [ABSTRACT FROM AUTHOR]

Symbiotic bacteria from seagrass and molluscs have the potential to be a source of secondary metabolites. Compounds from these secondary metabolites, such as natural food preservatives, can be used in the bioindustry sector. This research aims to identify the potential secondary metabolites from symbiotic bacteria in molluscs and seagrass that can support the Sustainable Development Goals (SDG) public health program. The samples used in this study were symbiotic bacteria of mollusc from seagrass environments, symbiotic bacteria from seagrass, and seagrass samples. The seagrass samples were analysed phytochemically, while the symbiotic bacteria samples from molluscs and seagrass were tested for their antibacterial activity against food spoilage pathogens such as Staphylococcus aureus and Escherichia coli using the disc diffusion method. Bacteria that showed potential antibacterial activity were identified using molecular methods, and the content of active compounds was analysed using gas chromatography-mass spectrometry. The phytochemical analysis revealed that seagrass contains flavones, tannins, steroids, and compounds with potential antibacterial properties. GC-MS analysis indicated that seagrass is predominantly composed of unsaturated fatty acids, which have antimicrobial effects by damaging bacterial cell membranes and disrupting the function of enzymes essential for growth and reproduction. Ten symbiotic bacteria isolates demonstrated proteolytic, cellulolytic, and lipolytic enzyme activity, suggesting their potential as antibacterial agents due to their ability to break down cell walls. Molecular identification of selected bacteria revealed the presence of mollusc symbionts, including Vibrio owensii, Bacillus paramycoides, and Pseudoalteromonas flavipulchra. The symbiotic bacteria found in seagrass in this study included Bacillus velezensis, Bacillus paramycoides, Nocardiopsis alba, Nocardiopsis alba, Bacillus tropicus, Lysinibacillus fusiformis, and Bacillus tropicus. GC-MS analysis of selected symbiotic bacteria from seagrass showed that the dominant compound components were phthalic acid, bis(7-methyl octyl) ester (25.18, 43.84, 35.55 %), and trimethoxy-3-piperidyl-2,2'-binaphthalene-1,1',4,4'-tetrone (36.36, 47.71, 64.48 %). In conclusion, seagrass and its symbiotic bacteria, as well as symbiotic bacteria from molluscs living in seagrass, have the potential to serve as antibacterial agents and can be used as references for food preservatives. [ABSTRACT FROM AUTHOR]

Bemisia tabaci is distributed globally and incurs considerable economic and ecological costs as an agricultural pest and viral vector. The entomopathogenic fungus Metarhizium anisopliae has been known for its insecticidal activity, but its impacts on whiteflies are understudied. We investigated how infection with the semi-persistently transmitted Cucurbit chlorotic yellows virus (CCYV) affects whitefly susceptibility to M. anisopliae exposure. We discovered that viruliferous whiteflies exhibited increased mortality when fungus infection was present compared to non-viruliferous insects. High throughput 16S rRNA sequencing also revealed significant alterations of the whitefly bacterial microbiome diversity and structure due to both CCYV and fungal presence. Specifically, the obligate symbiont Portiera decreased in relative abundance in viruliferous whiteflies exposed to M. anisopliae. Facultative Hamiltonella and Rickettsia symbionts exhibited variability across groups but dominated in fungus-treated non-viruliferous whiteflies. Our results illuminate triangular interplay between pest insects, their pathogens, and symbionts—dynamics which can inform integrated management strategies leveraging biopesticides This work underscores the promise of M. anisopliae for sustainable whitefly control while laying the groundwork for elucidating mechanisms behind microbe-mediated shifts in vector competence. [ABSTRACT FROM AUTHOR]

Objective: To investigate the symbiotic bacterial diversity of Hydra sinensis undergoing bleaching induced by strong light and high temperature. Method(s): The bleaching of H. sinensis was induced under strong light (6000 Lux, 25℃), high temperature (2000 Lux, 33℃) and combined stresses of strong light and high temperature (6000 Lux, 33℃), respectively. After 21 days of bleaching experiment, the bleaching degree of H. sinensis was detected by counting symbiotic algae in single cells of H. sinensis, and the diversity and abundance of symbiotic bacteria in H. sinensis were studied by use of high-throughput sequencing of 16S rDNA. Result(s): The number of symbiotic algae was observably decreased under strong light, high temperature and combined stresses of strong light and high temperature (P<0.05). The abundance of Phylum Cyanobacteria in H. sinensis increased under strong light, while the abundances of Phylum Cyanobacteria and Chlorobi in H. sinensis increased significantly under the combined stresses of strong light and high temperature. In addition, KEGG enrichment analysis indicated that symbiotic bacteria in H. sinensis undergoing bleaching were related to the biosynthesis and metabolism of antioxidants such as linoleic acid, carotenoid and betalain. Conclusion(s): Cyanobacteria and chloracea may provide their photosynthetic products to the host of H. sinensis to make up for the nutritional deficiency caused by the decrease of symbiotic algae. Furthermore, the diversity of symbiotic bacteria in H. sinensis changed in the direction of improving the antioxidant capacity of the host under strong light and/or high temperature stress. [ABSTRACT FROM AUTHOR]

Simple Summary: Acetamiprid, commonly used in agriculture to control aphids, faces the problem of resistance development in certain aphid species. To investigate the insect mechanisms involved in the adaptation process to acetamiprid, we compared the gene expression and microbial profiles of a pea aphid strain selected with acetamiprid (RS) with those of a non-selected strain (SS) using advanced omics techniques. The overall analysis revealed significant changes in the expression of genes involved in carbon and fatty acid metabolism in RS compared to those of SS aphids. In particular, we found increased expression of specific genes related to the synthesis of the components of the epidermal wax layer, suggesting that adaptation to acetamiprid involves the synthesis of a thicker protective layer. Additionally, subtle shifts in the bacterial composition of RS were detected. These results contribute valuable insights into the mechanisms underlying the pea aphid's response to acetamiprid exposure. Such understanding is essential for informing future research efforts and developing more effective strategies to control this pest. Acetamiprid is a broad-spectrum neonicotinoid insecticide used in agriculture to control aphids. While recent studies have documented resistance to acetamiprid in several aphid species, the underlying mechanisms are still not fully understood. In this study, we analyzed the transcriptome and metatranscriptome of a laboratory strain of the pea aphid, Acyrthosiphon pisum (Harris, 1776), with reduced susceptibility to acetamiprid after nine generations of exposure to identify candidate genes and the microbiome involved in the adaptation process. Sequencing of the transcriptome of both selected (RS) and non-selected (SS) strains allowed the identification of 14,858 genes and 4938 new transcripts. Most of the differentially expressed genes were associated with catalytic activities and metabolic pathways involving carbon and fatty acids. Specifically, alcohol-forming fatty acyl-CoA reductase (FAR) and acyl-CoA synthetase (ACSF2), both involved in the synthesis of epidermal wax layer components, were significantly upregulated in RS, suggesting that adaptation to acetamiprid involves the synthesis of a thicker protective layer. Metatranscriptomic analyses revealed subtle shifts in the microbiome of RS. These results contribute to a deeper understanding of acetamiprid adaptation by the pea aphid and provide new insights for aphid control strategies. [ABSTRACT FROM AUTHOR]

Due to global warming, the rice weevil is an increasing threat. Various species of weevils coexist with endosymbiotic Sodalis pierantonius. This study aimed to check whether different chemical properties of different cultivars of common wheat and barley grain cause disturbances in insect feeding and rearrangements of the rice weevil microbiome. It has been observed that foraging increases in the case of cultivars with low protein content, high steric acid content, and low antioxidant activity. Cultivars with higher antioxidant activity, a tendency was observed to increase the number of adult insects and the intensity of feeding. However, the correlation of number of offspring generations and antioxidants was different depending on species of cereals (R2=0.9 for barley, R2<0.1 for wheat). Changes of dominance classes of selected Enterobacteriaceae partially shifted the microbiomes of insects feeding on different barley varieties. In contrast, in some wheat cultivars, displacement of the dominant genus Sodalis by genera Staphylococcus or Mammaliicoccus was observed, severely reducing the rice weevil's foraging ability. Nevertheless, Sodalis sp. almost always dominated (from 16.7 to 90.07% for wheat and from 63.4 to 90.9% for barley), and no correlation was observed between species and variety factors of cereals and their abundance. [ABSTRACT FROM AUTHOR]

Abstract Mosquitoes are the deadliest animal in the word, transmitting a variety of insect-borne infectious diseases, such as malaria, dengue fever, yellow fever, and Zika, causing more deaths than any other vector-borne pathogen. Moreover, in the absence of effective drugs and vaccines to prevent and treat insect-borne diseases, mosquito control is particularly important as the primary measure. In recent decades, due to the gradual increase in mosquito resistance, increasing attention has fallen on the mechanisms and effects associated with pathogen infection. This review provides an overview of mosquito innate immune mechanisms in terms of physical and physiological barriers, pattern recognition receptors, signalling pathways, and cellular and humoral immunity, as well as the antipathogenic effects of mosquito symbiotic bacteria. This review contributes to an in-depth understanding of the interaction process between mosquitoes and pathogens and provides a theoretical basis for biological defence strategies against mosquito-borne infectious diseases. Graphical Abstract

Microorganisms colonize all barrier tissues and are present on the skin and all mucous membranes from birth. Bacteria have many ways of influencing the host organism, including activation of innate immunity receptors by pathogen-associated molecular patterns and synthesis of various chemical compounds, such as vitamins, short-chain fatty acids, bacteriocins, toxins. Bacteria, using extracellular vesicles, can also introduce high-molecular compounds, such as proteins and nucleic acids, into the cell, regulating the metabolic pathways of the host cells. Epithelial cells and immune cells recognize bacterial bioregulators and, depending on the microenvironment and context, determine the direction and intensity of the immune response. A large number of factors influence the maintenance of symbiotic microflora, the diversity of which protects hosts against pathogen colonization. Reduced bacterial diversity is associated with pathogen dominance and allergic diseases of the skin, gastrointestinal tract, and upper and lower respiratory tract, as seen in atopic dermatitis, allergic rhinitis, chronic rhinosinusitis, food allergies, and asthma. Understanding the multifactorial influence of microflora on maintaining health and disease determines the effectiveness of therapy and disease prevention and changes our food preferences and lifestyle to maintain health and active longevity.

Mosquitoes are the deadliest animal in the word, transmitting a variety of insect-borne infectious diseases, such as malaria, dengue fever, yellow fever, and Zika, causing more deaths than any other vector-borne pathogen. Moreover, in the absence of effective drugs and vaccines to prevent and treat insect-borne diseases, mosquito control is particularly important as the primary measure. In recent decades, due to the gradual increase in mosquito resistance, increasing attention has fallen on the mechanisms and effects associated with pathogen infection. This review provides an overview of mosquito innate immune mechanisms in terms of physical and physiological barriers, pattern recognition receptors, signalling pathways, and cellular and humoral immunity, as well as the antipathogenic effects of mosquito symbiotic bacteria. This review contributes to an in-depth understanding of the interaction process between mosquitoes and pathogens and provides a theoretical basis for biological defence strategies against mosquito-borne infectious diseases. [ABSTRACT FROM AUTHOR]

Insect-microbe endosymbiotic associations are omnipresent in nature, wherein the symbiotic microbes often play pivotal biological roles for their host insects. In particular, insects utilizing nutritionally imbalanced food sources are dependent on specific microbial symbionts to compensate for the nutritional deficiency via provisioning of B vitamins in blood-feeding insects, such as tsetse flies, lice, and bedbugs. Bat flies of the family Nycteribiidae (Diptera) are blood-sucking ectoparasites of bats and shown to be associated with co-speciating bacterial endosymbiont "Candidatus Aschnera chinzeii," although functional aspects of the microbial symbiosis have been totally unknown. In this study, we report the first complete genome sequence of Aschnera from the bristled bat fly Penicillidia jenynsii. The Aschnera genome consisted of a 748,020 bp circular chromosome and a 18,747 bp circular plasmid. The chromosome encoded 603 protein coding genes (including 3 pseudogenes), 33 transfer RNAs, and 1 copy of 16S/23S/5S ribosomal RNA operon. The plasmid contained 10 protein coding genes, whose biological function was elusive. The genome size, 0.77 Mbp, was drastically reduced in comparison with 4-6 Mbp genomes of free-living γ-proteobacteria. Accordingly, the Aschnera genome was devoid of many important functional genes, such as synthetic pathway genes for purines, pyrimidines, and essential amino acids. On the other hand, the Aschnera genome retained complete or near-complete synthetic pathway genes for biotin (vitamin B7), tetrahydrofolate (vitamin B9), riboflavin (vitamin B2), and pyridoxal 5'-phosphate (vitamin B6), suggesting that Aschnera provides these vitamins and cofactors that are deficient in the blood meal of the host bat fly. Similar retention patterns of the synthetic pathway genes for vitamins and cofactors were also observed in the endosymbiont genomes of other blood-sucking insects, such as Riesia of human lice, Arsenophonus of louse flies, and Wigglesworthia of tsetse flies, which may be either due to convergent evolution in the blood-sucking host insects or reflecting the genomic architecture of Arsenophonus-allied bacteria. [ABSTRACT FROM AUTHOR]

The ultrastructure of oogenic cells and mature oocytes has been described in a deep-sea clam, Calyptogena pacifica, for the first time. Specimens were collected in June 2018 from the Piip volcano in the Bering Sea (55.3821° N, 167.2613° E). Female germ cells developed in the branched gonadal tubules. The pattern of oogenic development is similar to that in littoral species. Although somatic accessory cells do not form any follicles around developing oocytes, these cells have a well-developed endoplasmic reticulum, which could be a source of yolk precursors and be involved in vitellogenesis. Lipid droplets appear in early oocytes, but they disappear later, suggesting that developing oocytes use these lipid droplets. At the onset of vitellogenesis, developing oocytes accumulate lipids in the ooplasm facing the gonadal tubule wall. Yolk granules, in contrast, appear at later stages of oogenesis. Mature oocytes, which are 200 µm in diameter, are filled with a significant volume of lipid droplets (occupying 24% of ooplasm) and a smaller volume of yolk granules (~ 8%). The mature oocyte's features, i.e., the large cell size (~ 200 µm in diameter) and the high abundance of ooplasmic inclusions, are typical of species with lecithotrophic development. The large volume of lipid droplets would enhance buoyancy of the oocytes and provide embryos and larvae with nutrients during dispersal. Symbiotic bacteria were found in accessory cells, in vitellogenic oocytes, and within remnants of degenerating oocytes. [ABSTRACT FROM AUTHOR]

Introduction: As tick-borne diseases rise to become the second most prevalent arthropod-transmitted disease globally, the increasing investigations focus on ticks correspondingly. Factors contributed to this increase include anthropogenic influences, changes in vertebrate faunal composition, social-recreational shifts, and climatic variation. Employing the 16S gene sequence method in next-generation sequencing (NGS) allows comprehensive pathogen identification in samples, facilitating the development of refined approaches to tick research omnidirectionally. Methods: In our survey, we compared the microbial richness and biological diversity of ticks in Wuwei City, Gansu province, differentiating between questing ticks found in grass and parasitic ticks collected from sheep based on 16S NGS method. Results: The results show Rickettsia, Coxiella, and Francisella were detected in all 50 Dermacentor nuttalli samples, suggesting that the co-infection may be linked to specific symbiotic bacteria in ticks. Our findings reveal significant differences in the composition and diversity of microorganisms, with the Friedmanniella and Bordetella genera existing more prevalent in parasitic ticks than in questing ticks (p  <  0.05). Additionally, the network analysis demonstrates that the interactions among bacterial genera can be either promotive or inhibitive in ticks exhibiting different lifestyles with the correlation index |r|  >  0.6. For instance, Francisella restrains the development of 10 other bacteria in parasitic ticks, whereas Phyllobacterium and Arthrobacter enhance colonization across all tick species. Discussion: By leveraging NGS techniques, our study reveals a high degree of species and phylogenetic diversity within the tick microbiome. It further highlights the potential to investigate the interplay between bacterial genera in both parasitic and questing ticks residing in identical habitat environments. [ABSTRACT FROM AUTHOR]

As an important fruit pest of global significance, Drosophila suzukii occupies a special ecological niche, with the characteristics of high sugar and low protein contents. This niche differs from those occupied by other fruit‐damaging Drosophila species. Gut bacteria substantially impact the physiology and ecology of insects. However, the contribution of gut microbes to the fitness of D. suzukii in their special ecological niche remains unclear. In this study, the effect of Klebsiella oxytoca on the development of D. suzukii was examined at physiological and molecular levels. The results showed that, after the removal of gut microbiota, the survival rate and longevity of axenic D. suzukii decreased significantly. Reintroduction of K. oxytoca to the midgut of D. suzukii advanced the development level of D. suzukii. The differentially expressed genes and metabolites between axenic and K. oxytoca‐reintroduced D. suzukii were enriched in the pathways of carbohydrate metabolism. This advancement was achieved through an increased glycolysis rate and the regulation of the transcript level of key genes in the glycolysis/gluconeogenesis pathway. Klebsiella oxytoca is likely to play an important role in increasing host fitness in their high‐sugar ecological niche by stimulating the glycolysis/gluconeogenesis pathway. As a protein source, bacteria can also provide direct nutrition for D. suzukii, which depends on the quantity or biomass of K. oxytoca. This result may provide a new target for controlling D. suzukii by inhibiting sugar metabolism through eliminating the effect of K. oxytoca and thus disrupting the balance of gut microbial communities. [ABSTRACT FROM AUTHOR]

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Abstract Background The true bugs (Heteroptera) occupy nearly all of the known ecological niches of insects. Among them, as a group containing more than 30,000 species, the phytophagous true bugs are making increasing impacts on agricultural and forestry ecosystems. Previous studies proved that symbiotic bacteria play important roles in these insects in fitting various habitats. However, it is still obscure about the evolutionary and ecological patterns of the microorganisms of phytophagous true bugs as a whole with comprehensive taxon sampling. Results Here, in order to explore the symbiotic patterns between plant-feeding true bugs and their symbiotic microorganisms, 209 species belonging to 32 families of 9 superfamilies had been sampled, which covered all the major phytophagous families of true bugs. The symbiotic microbial communities were surveyed by full-length 16S rRNA gene and ITS amplicons respectively for bacteria and fungi using the PacBio platform. We revealed that hosts mainly affect the dominant bacteria of symbiotic microbial communities, while habitats generally influence the subordinate ones. Thereafter, we carried out the ancestral state reconstruction of the dominant bacteria and found that dramatic replacements of dominant bacteria occurred in the early Cretaceous and formed newly stable symbiotic relationships accompanying the radiation of insect families. In contrast, the symbiotic fungi were revealed to be horizontally transmitted, which makes fungal communities distinctive in different habitats but not significantly related to hosts. Conclusions Host and habitat determine microbial communities of plant-feeding true bugs in different roles. The symbiotic bacterial communities are both shaped by host and habitat but in different ways. Nevertheless, the symbiotic fungal communities are mainly influenced by habitat but not host. These findings shed light on a general framework for future microbiome research of phytophagous insects. Video Abstract

Seed inoculation with symbiotic bacteria is a commonly employed practice in soybean cultivation. As a result, nodulation proceeds properly and plants self-supply atmospheric nitrogen, requiring either minimal or no additional nitrogen fertilization. The aim of the study was to investigate the response of soybeans to the application of the recommended or double dose of commercial inoculants (HiStick® Soy or TURBOSOY®) and/or mineral nitrogen fertilization compared to the untreated control. It was demonstrated that a double dose of the tested preparations had the most favorable effect on nodulation. However, the impact of weather conditions modified their effectiveness during the study years, which was especially visible in 2022. Sowing seeds without inoculation (control) resulted in the formation of sparse root nodules and consequently the lowest leaf area index (LAI) and soil plant analysis development (SPAD) measurements. In addition, the values of SPAD and LAI indices varied across the years of the study, indicating that weather conditions modified nitrogen uptake by plants. Overall, seed inoculation and/or nitrogen fertilization positively influenced the chemical composition of seeds compared to the control. The only decrease observed was in the oil content, while the double dose of HiStick® Soy preparation reduced the polyphenol content. The double dose of the tested inoculants had the most favorable impact on yield components and seed yield. However, applying inoculation at the recommended dose or in combination with nitrogen fertilization yielded similar or slightly worse results, depending on the year. Therefore, soybean seed inoculation should be recommended, although the effectiveness of the procedure will depend on various factors, including the type of inoculant, dosage, nitrogen fertilization, and weather conditions.

Secondary metabolites, bioactive compounds produced by living organisms, can unveil symbiotic relationships in nature. In this study, soilborne entomopathogenic nematodes associated with symbiotic bacteria (Xenorhabdus stockiae and Photorhabdus luminescens) were extracted from solvent supernatant containing secondary metabolites, demonstrating significant inhibitory effects against E. coli, S. aureus, B. subtilus, P. mirabilis, E. faecalis, and P. stutzeri. The characterization of these secondary metabolites by Fourier transforms infrared spectroscopy revealed amine groups of proteins, hydroxyl and carboxyl groups of polyphenols, hydroxyl groups of polysaccharides, and carboxyl groups of organic acids. Furthermore, the obtained crude extracts were analyzed by high-performance liquid chromatography for the basic identification of potential bioactive peptides. Gas chromatography–mass spectrometry analysis of ethyl acetate extracts from Xenorhabdus stockiae identified major compounds including nonanoic acid derivatives, proline, paromycin, octodecanal derivatives, trioxa-5-aza-1-silabicyclo, 4-octadecenal, methyl ester, oleic acid, and 1,2-benzenedicarboxylicacid. Additional extraction from Photorhabdus luminescens yielded functional compounds such as indole-3-acetic acid, phthalic acid, 1-tetradecanol, nemorosonol, 1-eicosanol, and unsaturated fatty acids. These findings support the potential development of novel natural antimicrobial agents for future pathogen suppression.

Meeting the demands of the growing population requires increased food and feed production, leading to higher levels of agri-food waste. As this type of waste seriously threatens public health and the environment, novel approaches to waste management should be developed. Insects have been proposed as efficient agents for biorefining waste, producing biomass that can be used for commercial products. However, challenges in achieving optimal outcomes and maximizing beneficial results remain. Microbial symbionts associated with insects are known to have a critical role in the development, fitness, and versatility of insects, and as such, they can be utilized as targets for the optimization of agri-food waste insect-based biorefinery systems. This review discusses insect-based biorefineries, focusing on the agricultural applications of edible insects, mainly as animal feed and organic fertilizers. We also describe the interplay between agri-food waste-utilizing insects and associated microbiota and the microbial contribution in enhancing insect growth, development, and involvement in organic waste bioconversion processes. The potential contribution of insect gut microbiota in eliminating pathogens, toxins, and pollutants and microbe-mediated approaches for enhancing insect growth and the bioconversion of organic waste are also discussed. The present review outlines the benefits of using insects in agri-food and organic waste biorefinery systems, describes the roles of insect-associated microbial symbionts in waste bioconversion processes, and highlights the potential of such biorefinery systems in addressing the current agri-food waste-related challenges.

Background: The true bugs (Heteroptera) occupy nearly all of the known ecological niches of insects. Among them, as a group containing more than 30,000 species, the phytophagous true bugs are making increasing impacts on agricultural and forestry ecosystems. Previous studies proved that symbiotic bacteria play important roles in these insects in fitting various habitats. However, it is still obscure about the evolutionary and ecological patterns of the microorganisms of phytophagous true bugs as a whole with comprehensive taxon sampling. Results: Here, in order to explore the symbiotic patterns between plant-feeding true bugs and their symbiotic microorganisms, 209 species belonging to 32 families of 9 superfamilies had been sampled, which covered all the major phytophagous families of true bugs. The symbiotic microbial communities were surveyed by full-length 16S rRNA gene and ITS amplicons respectively for bacteria and fungi using the PacBio platform. We revealed that hosts mainly affect the dominant bacteria of symbiotic microbial communities, while habitats generally influence the subordinate ones. Thereafter, we carried out the ancestral state reconstruction of the dominant bacteria and found that dramatic replacements of dominant bacteria occurred in the early Cretaceous and formed newly stable symbiotic relationships accompanying the radiation of insect families. In contrast, the symbiotic fungi were revealed to be horizontally transmitted, which makes fungal communities distinctive in different habitats but not significantly related to hosts. Conclusions: Host and habitat determine microbial communities of plant-feeding true bugs in different roles. The symbiotic bacterial communities are both shaped by host and habitat but in different ways. Nevertheless, the symbiotic fungal communities are mainly influenced by habitat but not host. These findings shed light on a general framework for future microbiome research of phytophagous insects. DpRsUfddvNEYPX-MDhm9d6 Video Abstract [ABSTRACT FROM AUTHOR]

Many insects are associated with endosymbionts that influence the feeding, reproduction, and distribution of their hosts. Although the small green mirid, Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae), a zoophytophagous predator that feeds on plants as well as arthropods, is a globally important biological control agent, its microbiome has not been sufficiently studied. In the present study, we assessed the microbiome variation in 96 N. tenuis individuals from 14 locations throughout Japan, based on amplicon sequencing of the 16S ribosomal RNA gene. Nine major bacteria associated with N. tenuis were identified: Rickettsia, two strains of Wolbachia, Spiroplasma, Providencia, Serratia, Pseudochrobactrum, Lactococcus, and Stenotrophomonas. Additionally, a diagnostic PCR analysis for three typical insect reproductive manipulators, Rickettsia, Wolbachia, and Spiroplasma, was performed on a larger sample size (n = 360) of N. tenuis individuals; the most prevalent symbiont was Rickettsia (69.7%), followed by Wolbachia (39.2%) and Spiroplasma (6.1%). Although some symbionts were co-infected, their prevalence did not exhibit any specific tendency, such as a high frequency in specific infection combinations. The infection frequency of Rickettsia was significantly correlated with latitude and temperature, while that of Wolbachia and Spiroplasma was significantly correlated with host plants. The predominance of these bacteria and the absence of obligate symbionts suggested that the N. tenuis microbiome is typical for predatory arthropods rather than sap-feeding insects. Rickettsia and Wolbachia were vertically transmitted rather than horizontally transmitted from the prey. The functional validation of each symbiont would be warranted to develop N. tenuis as a biological control agent. [ABSTRACT FROM AUTHOR]

Multicellular organisms live in environments containing diverse nutrients and a wide variety of microbial communities. On the one hand, the immune response of organisms can protect from the intrusion of exogenous microorganisms. On the other hand, the dynamic coordination of anabolism and catabolism of organisms is a necessary factor for growth and reproduction. Since the production of an immune response is an energy-intensive process, the activation of immune cells is accompanied by metabolic transformations that enable the rapid production of ATP and new biomolecules. In insects, the coordination of immunity and metabolism is the basis for insects to cope with environmental challenges and ensure normal growth, development and reproduction. During the activation of insect immune tissues by pathogenic microorganisms, not only the utilization of organic resources can be enhanced, but also the activated immune cells can usurp the nutrients of non-immune tissues by generating signals. At the same time, insects also have symbiotic bacteria in their body, which can affect insect physiology through immunemetabolic regulation. This paper reviews the research progress of insect immune-metabolism regulation from the perspective of insect tissues, such as fat body, gut and hemocytes. The effects of microorganisms (pathogenic bacteria/non-pathogenic bacteria) and parasitoids on immune-metabolism were elaborated here, which provide guidance to uncover immunometabolism mechanisms in insects and mammals. This work also provides insights to utilize immune-metabolism for the formulation of pest control strategies. [ABSTRACT FROM AUTHOR]

Background: Commensal microorganisms are widely distributed in insect gut tissues and play important roles in host nutrition, metabolism, reproductive regulation, and especially immune functioning and tolerance to pathogens. Consequently, gut microbiota represent a promising resource for the development of microbial‐based products for pest control and management. However, the interactions among host immunity, entomopathogen infections, and gut microbiota remain poorly understood for many arthropod pests. Results: We previously isolated an Enterococcus strain (HcM7) from Hyphantria cunea larvae guts that increased the survival rates of larvae challenged with nucleopolyhedrovirus (NPV). Here, we further investigated whether this Enterococcus strain stimulates a protective immune response against NPV proliferation. Infection bioassays demonstrated that re‐introduction of the HcM7 strain to germfree larvae preactivated the expression of several antimicrobial peptides (particularly H. cunea gloverin 1, HcGlv1), resulting in the significant repression of virus replication in host guts and hemolymph, and consequently improved host survivorship after NPV infection. Furthermore, silencing of the HcGlv1 gene by RNA interference markedly enhanced the deleterious effects of NPV infection, revealing a role of this gut symbiont‐induced gene in host defenses against pathogenic infections. Conclusion: These results show that some gut microorganisms can stimulate host immune systems, thereby contributing to resistance to entomopathogens. Furthermore, HcM7, as a functional symbiotic bacteria of H. cunea larvae, may be a potential target for increasing the effectiveness of biocontrol agents against this devastating pest. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Abstract Background Neonicotinoid insecticides are applied worldwide for the control of agricultural insect pests. The evolution of neonicotinoid resistance has led to the failure of pest control in the field. The enhanced detoxifying enzyme activity and target mutations play important roles in the resistance of insects to neonicotinoid resistance. Emerging evidence indicates a central role of the gut symbiont in insect pest resistance to pesticides. Existing reports suggest that symbiotic microorganisms could mediate pesticide resistance by degrading pesticides in insect pests. Results The 16S rDNA sequencing results showed that the richness and diversity of the gut community between the imidacloprid-resistant (IMI-R) and imidacloprid-susceptible (IMI-S) strains of the cotton aphid Aphis gossypii showed no significant difference, while the abundance of the gut symbiont Sphingomonas was significantly higher in the IMI-R strain. Antibiotic treatment deprived Sphingomonas of the gut, followed by an increase in susceptibility to imidacloprid in the IMI-R strain. The susceptibility of the IMI-S strain to imidacloprid was significantly decreased as expected after supplementation with Sphingomonas. In addition, the imidacloprid susceptibility in nine field populations, which were all infected with Sphingomonas, increased to different degrees after treatment with antibiotics. Then, we demonstrated that Sphingomonas isolated from the gut of the IMI-R strain could subsist only with imidacloprid as a carbon source. The metabolic efficiency of imidacloprid by Sphingomonas reached 56% by HPLC detection. This further proved that Sphingomonas could mediate A. gossypii resistance to imidacloprid by hydroxylation and nitroreduction. Conclusions Our findings suggest that the gut symbiont Sphingomonas, with detoxification properties, could offer an opportunity for insect pests to metabolize imidacloprid. These findings enriched our knowledge of mechanisms of insecticide resistance and provided new symbiont-based strategies for control of insecticide-resistant insect pests with high Sphingomonas abundance.

Abstract Background Apolygus lucorum is a worldwide omnivorous pest damaging a range of crops and causing great economic losses. Symbiotic bacteria living in insects play a key role in the nutrition, physiology, and behavior of hosts. Here, we present an experiment using Illumina HiSeq sequencing targeting the V3–V4 regions of bacteria’s 16S rRNA throughout the entire life cycle of A. lucorum. Results The first and second instar nymphs have the largest alpha diversity compared with other life stages of the insect. Bacterial phyla Proteobacteria (72.29%), Firmicutes (15.24%), Actinobacteria (7.76%) exhibit the largest relative abundance in all developmental stages. Erwinia (23.97%) and Lactococcus (10.62%) are the two genera with the highest relative abundance. The relative abundance of Erwinia in the nymph stage is significantly greater than the adult stage, and the relative abundance of Lactococcus in 6-day-old and 9-day-old adult females is higher compared with adult males. Conclusions These results reveal that microbial community composition and relative abundance shift dynamically at different life stages, implying that different bacterial phyla and genera may have specific roles in specific life stages such as metabolism, nutrition absorption, detoxification, and reproduction. This study reveals for the first time the community composition and ecological dynamics of symbiotic bacteria throughout the life stages of A. lucorum, and thus may provide insight to new strategies for pest control.

Bemisia tabaci is distributed globally and incurs considerable economic and ecological costs as an agricultural pest and viral vector. The entomopathogenic fungus Metarhizium anisopliae has been known for its insecticidal activity, but its impacts on whiteflies are understudied. We investigated how infection with the semi-persistently transmitted Cucurbit chlorotic yellows virus (CCYV) affects whitefly susceptibility to M. anisopliae exposure. We discovered that viruliferous whiteflies exhibited increased mortality when fungus infection was present compared to non-viruliferous insects. High throughput 16S rRNA sequencing also revealed significant alterations of the whitefly bacterial microbiome diversity and structure due to both CCYV and fungal presence. Specifically, the obligate symbiont Portiera decreased in relative abundance in viruliferous whiteflies exposed to M. anisopliae. Facultative Hamiltonella and Rickettsia symbionts exhibited variability across groups but dominated in fungus-treated non-viruliferous whiteflies. Our results illuminate triangular interplay between pest insects, their pathogens, and symbionts—dynamics which can inform integrated management strategies leveraging biopesticides This work underscores the promise of M. anisopliae for sustainable whitefly control while laying the groundwork for elucidating mechanisms behind microbe-mediated shifts in vector competence.

Acetamiprid is a broad-spectrum neonicotinoid insecticide used in agriculture to control aphids. While recent studies have documented resistance to acetamiprid in several aphid species, the underlying mechanisms are still not fully understood. In this study, we analyzed the transcriptome and metatranscriptome of a laboratory strain of the pea aphid, Acyrthosiphon pisum (Harris, 1776), with reduced susceptibility to acetamiprid after nine generations of exposure to identify candidate genes and the microbiome involved in the adaptation process. Sequencing of the transcriptome of both selected (RS) and non-selected (SS) strains allowed the identification of 14,858 genes and 4938 new transcripts. Most of the differentially expressed genes were associated with catalytic activities and metabolic pathways involving carbon and fatty acids. Specifically, alcohol-forming fatty acyl-CoA reductase (FAR) and acyl-CoA synthetase (ACSF2), both involved in the synthesis of epidermal wax layer components, were significantly upregulated in RS, suggesting that adaptation to acetamiprid involves the synthesis of a thicker protective layer. Metatranscriptomic analyses revealed subtle shifts in the microbiome of RS. These results contribute to a deeper understanding of acetamiprid adaptation by the pea aphid and provide new insights for aphid control strategies.

The Japanese pine sawyer, Monochamus saltuarius, as a beetle vector of Bursaphelenchus xylophilus (pine wood nematode), is an economically important forest pest in Eurasia. To feed on the phloem and xylem of conifers, M. saltuarius needs to overcome various stress factors, including coping with entomopathogenic bacteria and also various plant secondary compounds (PSCs). As an important adaptation strategy to colonize host trees, M. saltuarius deposit eggs in oviposition pits to shield their progeny. These pits harbor bacterial communities that are involved in the host adaptation of M. saltuarius to the conifers. However, the composition, origin, and functions of these oviposition pit bacteria are rarely understood. In this study, we investigated the bacterial community associated with M. saltuarius oviposition pits and their ability to degrade PSCs. Results showed that the bacterial community structure of M. saltuarius oviposition pits significantly differed from that of uninfected phloem. Also, the oviposition pit bacteria were predicted to be enriched in PSC degradation pathways. The microbial community also harbored a lethal strain of Serratia, which was significantly inhibited. Meanwhile, metatranscriptome analysis indicated that genes involved in PSCs degradation were expressed complementarily among the microbial communities of oviposition pits and secretions. In vitro degradation showed that bacteria cultured from oviposition pits degraded more monoterpenes and flavonoids than bacteria cultured from uninfected phloem isolates. Disinfection of oviposition pits increased the mortality of newly hatched larvae and resulted in a significant decrease in body weight in the early stages. Overall, our results reveal that M. saltuarius construct oviposition pits that harbor a diverse microbial community, with stronger PSCs degradation abilities and a low abundance of entomopathogenic bacteria, resulting in the increased fitness of newly hatched larvae. [ABSTRACT FROM AUTHOR]

Organophosphate insecticides have been widely used in agriculture to control insect pests and raised growing problems including insecticide resistance. According to recent studies, microbial symbionts play important roles in developing insecticide resistance. The bean bug Riptortus pedestris is a notorious pest of leguminous crops, harbouring diverse microorganisms in their insect midguts. However, the potential biological roles of many gut‐associated bacteria are largely unknown. In this study, we isolated a bacterial strain from the midgut of R. pedestris and identified it as Serratia marcescens based on its morphological characteristics and the phylogenetic analysis of the 16S rRNA gene sequence. A halo of phosphorus‐solubilizing bacteria was observed on a medium containing lecithin and an organophosphorus insecticide dimethoate. Subsequently, the microbial degradation of dimethoate was determined using Gas Chromatography with Mass Spectroscopy (GC–MS). Further analysis indicated that the S. marcescens contained organophosphorus‐degrading MBL‐fold metallo‐hydrolase gene. Additionally, we discovered that the bacterium could colonize the insect midgut stably and enhance the host survivorship when exposed to dimethoate. Our results revealed that an insect gut‐associated bacterium could degrade organophosphorus compounds, thereby reducing related insecticide toxicity to the insect hosts. These findings provided further insights into the host‐microbiota interactions underpinning the development of insecticide resistance. [ABSTRACT FROM AUTHOR]

Among hundreds of insect families, Hermatobatidae (commonly known as coral treaders) is one of the most unique. They are small, wingless predaceous bugs in the suborder Heteroptera. Adults are almost black in colour, measuring about 5 mm in body length and 3 mm in width. Thirteen species are known from tropical coral reefs or rocky shores, but their origin and evolutionary adaptation to their unusual marine habitat were unexplored. We report here the genome and metagenome of Hermatobates lingyangjiaoensis, hitherto known only from its type locality in the South China Sea. We further reconstructed the evolutionary history and origin of these marine bugs in the broader context of Arthropoda. The dated phylogeny indicates that Hexapoda diverged from their marine sister groups approximately 498 Ma and that Hermatobatidae originated 192 Ma, indicating that they returned to an oceanic life some 300 Myr after their ancestors became terrestrial. Their origin is consistent with the recovery of tropical reef ecosystems after the end-Triassic mass extinction, which might have provided new and open niches for them to occupy and thrive. Our analyses also revealed that both the genome changes and the symbiotic bacteria might have contributed to adaptations necessary for life in the sea. [ABSTRACT FROM AUTHOR]

The symbiotic bacteria–insect interaction is considered to be associated with immunity and drug resistance. However, the wide variety of insect species and habitats is thought to have a significant impact on the symbiotic community, leading to disparate results. Here, we demonstrated that symbiotic bacteria regulated the immune response by changing the proportion of the Gram-positive and the Gram-negative bacterial community in Lymantria dispar (L. dispar) after infection with its viral pathogen, L. dispar Nucleopolyhedrovirus (LdMNPV). After oral infection, the immune deficiency pathway was activated immediately, and the expression of Relish was up-regulated to promote the secretion of antimicrobial peptides. Meanwhile, the abundance of the Gram-negative bacterial community increased at the same time. Moreover, the Toll pathway was not regulated in the same way as the Imd pathway was after infection. However, the change in the Toll pathway's expression remained positively correlated to the abundance of Gram-positive bacteria. This finding implied that the ratio of Gram-negative to Gram-positive bacteria in the LdMNPV infected larvae had an effect on the immune response. Our findings revealed that the immune regulation of L. dispar was regulated by the relative abundance of its symbiotic bacteria at different infection times with LdMNPV, which provides a new way to understand symbiotic bacteria–insect interactions. [ABSTRACT FROM AUTHOR]

The gut microbiome of bees is vital for the health of their hosts. Given the ecosystem functions performed by bees, and the declines faced by many species, it is important to improve our understanding of the amount of natural variation in the gut microbiome, the level of sharing of bacteria among co-occurring species (including between native and non-native species), and how gut communities respond to infections. We conducted 16S rRNA metabarcoding to discern the level of microbiome similarity between honey bees (Apis mellifera, N = 49) and bumble bees (Bombus spp., N = 66) in a suburban-rural landscape. We identified a total of 233 amplicon sequence variants (ASVs) and found simple gut microbiomes dominated by bacterial taxa belonging to Gilliamella, Snodgrassella, and Lactobacillus. The average number of ASVs per species ranged from 4.00-15.00 (8.79 ± 3.84, mean ± SD). Amplicon sequence variant of one bacterial species, G. apicola (ASV 1), was widely shared across honey bees and bumble bees. However, we detected another ASV of G. apicola that was either exclusive to honey bees, or represented an intra-genomic 16S rRNA haplotype variant in honey bees. Other than ASV 1, honey bees and bumble bees rarely share gut bacteria, even ones likely derived from outside environments (e.g., Rhizobium spp., Fructobacillus spp.). Honey bee bacterial microbiomes exhibited higher alpha diversity but lower beta and gamma diversities than those of bumble bees, likely a result of the former possessing larger, perennial hives. Finally, we identified pathogenic or symbiotic bacteria (G. apicola, Acinetobacter sp. and Pluralibacter sp.) that associate with Trypanosome and/or Vairimorpha infections in bees. Such insights help to determine bees' susceptibility to infections should gut microbiomes become disrupted by chemical pollutants and contribute to our understanding of what constitutes a state of dysbiosis. [ABSTRACT FROM AUTHOR]