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Key Message The incorporation of antibiotics to field-collected Chilo suppressalis larvae significantly reduce bacterial quantities and affect larval performance on rice plants. Field-collected Chilo suppressalis larvae deposit abundant bacteria onto feeding sites through oral secretion, which modulate defense responses in rice plants. Bacteria isolated from larval oral secretion (such as Enterobacter and Acinetobacter) contribute to preventing rice plants to fully activate jasmonic acid-mediated antiherbivore defenses. Our results indicate insect-associated bacteria play an important role for C. suppressalis to adapt to rice plants, which could also be treated as targets for developing novel pest management strategies.

The coastal areas of the Red Sea within Saudi Arabia have a wide diversity of marine soft corals harboring many symbiotic bacteria that could present a potential source of many antibacterial metabolites. This study focused on the molecular characterization of antibiotic-producing bacteria associated with soft corals and investigated the influence of culture conditions on the production of their antibacterial extracts. Two soft coral samples were collected offshore from Yanbu City and identified as genus Sarcophyton. Screening for antibiotic-producing bacterial symbionts uncovered nine isolates with a wide inhibitory spectrum against gram-positive and negative bacteria. Phylogenetic analysis of the bioactive bacterial symbionts 16S rRNA gene sequences revealed the prevalence of gram-negative bacteria with seven isolates all belonging to class γ-proteobacteria. The remaining two isolates were represented within class firmicutes and actinobacteria. Eight isolates were grouped perfectly with their best matches. While the remaining ninth isolate was phylogenetically separated from all the species within the Photobacterium genus, suggesting that it could be a novel species. The optimum antibacterial compounds production was obtained in minimal media, at a temperature range of 25–28 °C, pH 7 and 100–150 rev/min agitation during 24–48 h of the incubation period, with moderate to high degree of thermal stability.

Abstract Background The tomato leaf miner, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a major pest of tomato plants threatening global tomato production. The control of the pest is becoming increasingly difficult due to the rapid development of resistance to insecticides. Entomopathogenic nematodes (EPNs) of families Steinernematidae and Heterorhabditidae are successful biocontrol agents for many insect pests. Recently, their bacteria, Xenorhabdus spp. and Photorhabdus spp. have attracted great attention due to their major role in the pathogenicity of EPNs. In the present study, the pathogenicity of eleven EPNs isolates belonging to Steinernema feltiae and Heterorhabditis bacteriophora species was screened against the 1st/2nd and 3rd/4th instar larvae of T. absoluta. The cell-free supernatants and cell suspensions of the symbiotic bacteria from the most efficient isolates were further evaluated for their biocontrol potential in the oral and contact treatments on the larvae of T. absoluta. Results KBC-4 and MCB-8 isolates of S. feltiae showed superior virulence relative to other EPNs species/isolates and induced 90% larval mortality against the 3rd/4th instar of T. absoluta larvae, whereas there were no clear differences in the efficacy of EPNs species/isolates against the 1st/2nd instar of T. absoluta. The 1st/2nd instar of T. absoluta larvae was more susceptible to cell-free supernatants and cell suspension of selected EPNs. The highest mortality (80%) was obtained from X. bovienii MCB-8 strain in the contact treatment of supernatants. In contact treatment of cell suspension, higher mortalities were obtained compared to oral treatments. Mortality rates ranged between 30 and 57.5% in the contact treatments of cell suspensions while the highest mortality did not exceed 20% in oral treatments. The antifeedant activity was observed in oral treatments of cell suspension and most of the larvae avoided feeding on treated leaves. Conclusion The results indicated that symbiotic bacteria of EPNs had a great potential against T. absoluta larvae and contact treatment of cell-free supernatants against early instars of T. absoluta can be an ideal application. However, further studies are needed to investigate the field effectiveness of symbiotic bacteria.

Planthopper infestation in rice causes direct and indirect damage through feeding and viral transmission. Host microbes and small RNAs (sRNAs) play essential roles in regulating biological processes, such as metabolism, development, immunity, and stress responses in eukaryotic organisms, including plants and insects. Recently, advanced metagenomic approaches have facilitated investigations on microbial diversity and its function in insects and plants, highlighting the significance of microbiota in sustaining host life and regulating their interactions with the environment. Recent research has also suggested significant roles for sRNA-regulated genes during rice–planthopper interactions. The response and behavior of the rice plant to planthopper feeding are determined by changes in the host transcriptome, which might be regulated by sRNAs. In addition, the roles of microbial symbionts and sRNAs in the host response to viral infection are complex and involve defense-related changes in the host transcriptomic profile. This review reviews the structure and potential functions of microbes and sRNAs in rice and the associated planthopper species. In addition, the involvement of the microbiota and sRNAs in the rice–planthopper–virus interactions during planthopper infestation and viral infection are discussed.

Simple Summary: Acyrthosiphon gossypii is a sap-sucking pest that causes direct cotton damage by sucking sap and indirect damage by honeydew contamination and induced fungal growth. Chemical pesticides are widely used to combat the damage caused by A. gossypii. However, the escalating use of pesticides has had a notable impact on the resurgence and resistance of pests. In addition, microbes that live in insects as symbionts have recently been found to protect their hosts against toxins. In this study, we determined the toxicity and sublethal effects of the neonicotinoid insecticide imidacloprid on A. gossypii and further determined the effects of imidacloprid on symbiotic bacteria. The sublethal concentrations of imidacloprid had a negative effect on the reproduction and longevity of the G0 generation of A. gossypii, increased the population growth parameters of the G1 and G2 generations, but not those of the G3 generation, and altered the abundance of symbiotic bacteria. Our results revealed the transgenerational and sublethal effects of imidacloprid on A. gossypii, which may have implications for the endosymbiont-insecticide of the major pest. Symbiotic bacteria and hormesis in aphids are the driving forces for pesticide resistance. However, the mechanism remains unclear. In this study, the effects of imidacloprid on the population growth parameters and symbiotic bacterial communities of three successive generations of Acyrthosiphon gossypii were investigated. The bioassay results showed that imidacloprid had high toxicity to A. gossypii with an LC50 of 1.46 mg·L−1. The fecundity and longevity of the G0 generation of A. gossypii decreased when exposed to the LC15 of imidacloprid. The net reproductive rate (R0), intrinsic rate of increase (rm), finite rate of increase (λ), and total reproductive rate (GRR) of G1 and G2 offspring were significantly increased, but those of the control and G3 offspring were not. In addition, sequencing data showed that the symbiotic bacteria of A. gossypii mainly belonged to Proteobacteria, with a relative abundance of 98.68%. The dominant genera of the symbiotic bacterial community were Buchnera and Arsenophonus. After treatment with the LC15 of imidacloprid, the diversity and species number of bacterial communities of A. gossypii decreased for G1–G3 and the abundance of Candidatus-Hamiltonella decreased, but Buchnera increased. These results provide insight into the resistance mechanism of insecticides and the stress adaptation between symbiotic bacteria and aphids. [ABSTRACT FROM AUTHOR]

Camellia spiny whitefly, Aleurocanthus camelliae (Hemiptera: Aleyrodidae), is a major pest in tea, which poses a serious threat to tea production. Similar to many insects, various bacterial symbioses inside A. camelliae may participate in the reproduction, metabolism, and detoxification of the host. However, few reports included research on the microbial composition and influence on A. camelliae growth. We first applied high-throughput sequencing of the V4 region in the 16S rRNA of symbiotic bacteria to study its component and effect on the biological trait of A. camelliae by comparing it with the antibiotic treatment group. The population parameters, survival rate, and fecundity rate of A. camelliae were also analyzed using the age--stage two-sex life table. Our results demonstrated that phylum Proteobacteria (higher than 96.15%) dominated the whole life cycle of A. camelliae. It unveiled the presence of Candidatus Portiera (primary endosymbiont) (67.15-73.33%), Arsenophonus (5.58-22.89%), Wolbachia (4.53-11.58%), Rickettsia (0.75-2.59%), and Pseudomonas (0.99-1.88%) genus. Antibiotic treatment caused a significant decrease in the endosymbiont, which negatively affected the host's biological properties and life process. For example, 1.5% rifampicin treatment caused a longer preadult stage in the offspring generation (55.92 d) compared to the control (49.75d) and a lower survival rate (0.36) than the control (0.60). The decreased intrinsic rate of increase (r), net reproductive rate (R0), and prolonged mean generation time (T) were signs of all disadvantageous effects associated with symbiotic reduction. Our findings confirmed the composition and richness of symbiotic bacteria in larva and adult of A. camelliae by an Illumina NovaSeq 6000 analysis and their influence on the development of the host by demographic research. Together, the results suggested that symbiotic bacteria play an important role inmanipulating the biological development of their hosts, whichmight help us for developing new pest control agents and technologies for better management of A. camelliae. [ABSTRACT FROM AUTHOR]

Symbiotic bacteria form a mutualistic relationship with nematodes and are pathogenic to many insect pests. They kill insects using various strategies to evade or suppress their humoral and cellular immunity. Here we evaluate the toxic effects of these bacteria and their secondary metabolites on the survival and phenoloxidase (PO) activation of Octodonta nipae larvae using biochemical and molecular methods. The results show P. luminescens H06 and X. nematophila All treatments caused significant reductions in the number of O. nipae larvae in a dose-dependent manner. Secondly, the O. nipae immune system recognizes symbiotic bacteria at early and late stages of infection via the induction of C-type lectin. Live symbiotic bacteria significantly inhibit PO activity in O. nipae whereas heat-treated bacteria strongly increase PO activity. Additionally, expression levels of four O. nipae proPhenoloxidase genes following treatment with P. luminescens H06 and X. nematophila All were compared. We found that the expression levels of all proPhenoloxidase genes were significantly down-regulated at all-time points. Similarly, treatments of O. nipae larvae with metabolites benzylideneacetone and oxindole significantly down-regulated the expression of the PPO gene and inhibited PO activity. However, the addition of arachidonic acid to metabolite-treated larvae restored the expression level of the PPO gene and increased PO activity. Our results provide new insight into the roles of symbiotic bacteria in countering the insect phenoloxidase activation system. [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.

While genetically modified (GM) crops bring economic benefits to human beings, their impact on non-target organisms has become an important part of environmental safety assessments. Symbiotic bacteria play an important role in eukaryotic biological functions and can adjust host communities to adapt to new environments. Therefore, this study examined the effects of Cry1B protein on the growth and development of non-target natural enemies of Pardosa astrigera (L. Koch) from the perspective of symbiotic bacteria. Cry1B protein had no significant effect on the health indicators of P. astrigera (adults and 2nd instar spiderlings). 16S rRNA sequencing results revealed that Cry1B protein did not change the symbiotic bacteria species composition of P. astrigera, but did reduce the number of OTU and species diversity. In 2nd instar spiderlings, neither the dominant phylum (Proteobacteria) nor the dominant genus (Acinetobacter) changed, but the relative abundance of Corynebacterium-1 decreased significantly; in adult spiders, the dominant bacteria genera of females and males were different. The dominant bacterial genera were Brevibacterium in females and Corynebacterium-1 in males, but Corynebacterium-1 was the dominant bacteria in both females and males feeding on Cry1B. The relative abundance of Wolbachia also increased significantly. In addition, bacteria in other genera varied significantly by sex. KEGG results showed that Cry1B protein only altered the significant enrichment of metabolic pathways in female spiders. In conclusion, the effects of Cry1B protein on symbiotic bacteria vary by growth and development stage and sex.

Antarctic microbes are important agents for evolutionary adaptation and natural resource of bioactive compounds, harboring the particular metabolic pathways to biosynthesize natural products. However, not much is known on symbiotic microbiomes of fish in the Antarctic zone. In the present study, the culture method and whole-genome sequencing were performed. Natural product analyses were carried out to determine the biosynthetic potential. We report the isolation and identification of a symbiotic bacterium Serratia myotis L7-1, that is highly adaptive and resides within Antarctic fish, Trematomus bernacchii. As revealed by genomic analyses, Antarctic strain S. myotis L7-1 possesses carbohydrate-active enzymes (CAZymes), biosynthetic gene clusters (BGCs), stress response genes, antibiotic resistant genes (ARGs), and a complete type IV secretion system which could facilitate competition and colonization in the extreme Antarctic environment. The identification of microbiome gene clusters indicates the biosynthetic potential of bioactive compounds. Based on bioactivity-guided fractionation, serranticin was purified and identified as the bioactive compound, showing significant antibacterial and antitumor activity. The serranticin gene cluster was identified and located on the chrome. Furthermore, the multidrug resistance and strong bacterial antagonism contribute competitive advantages in ecological niches. Our results highlight the existence of a symbiotic bacterium in Antarctic fish largely represented by bioactive natural products and the adaptability to survive in the fish living in Antarctic oceans. [ABSTRACT FROM AUTHOR]

Introduction: Reproductive polymorphism and symbiotic bacteria are commonly observed in aphids, but their interaction remains largely unclear. In polymorphic aphid species (Aphis gossypii), offspring of parthenogenetic females (PFs) develops into sexuparae which produces gynoparae and males successively. Gynoparae further produces sexual females (SFs), and these sexual females mate with males to produce offspring. Methods: In this study, we investigated the dynamic changes of symbiotic bacteria during the above-mentioned five reproductive morph switch in A. gossypii via 16S rRNA sequencing technology. Results: The results showed that species richness and community diversity of symbiotic bacteria in males were the highest. Proteobacteria was absolutely dominant bacterial phylum (with relative abundance of more than 90%) in the five reproductive morphs of A. gossypii, and Buchnera was absolutely dominant genus (with relative abundance of >90%), followed by Rhodococcus, Pseudomonas, and Pantoea. Male-killing symbiont Arsenophonus presented the highest relative abundance in gynoparae, a specific morph whose offsprings were exclusively sexual females. Both principal component analysis (PCA) and clustering analysis showed trans-generation similarity in microbial community structure between sexuparae and sexual females, between PFs and gynoparae. PICRUSt 2 analysis showed that symbiotic bacteria in the five reproductive morphs were mainly enriched in metabolic pathways. Discussion: Reproductive morph switch induced by environmental changes might be associated with bacterial community variation and sexual polymorphism of aphids. This study provides a new perspective for further deciphering the interactions between microbes and reproductive polymorphism in host aphids. [ABSTRACT FROM AUTHOR]

Plant defenses in response to chewing insects are generally regulated by jasmonic acid (JA) signaling pathway, whereas salicylic acid (SA) signaling is mainly involved in plant defense against biotrophic pathogens and piercing-sucking insects. Previous studies showed that both JA- and SA-related defenses in rice plants were triggered by the infestation of the rice striped stem borer (SSB, Chilo suppressalis), a destructive pest causing severe damage to rice production. Herbivore-associated microbes play an important role in modulating plant–insect interaction, and thus we speculate that the SSB symbiotic microbes acting as a hidden player may cause this anomalous result. The antibiotics (AB) treatment significantly depressed the performance of field-collected SSB larvae on rice plants, and reduced the quantities of bacteria around the wounds of rice stems compared to non-AB treatment. In response to mechanical wounding and oral secretions (OS) collected from non-AB treated larvae, rice plants exhibited lower levels of JA-regulated defenses, but higher levels of SA-regulated defenses compared to the treatment of OS from AB-treated larvae determined by using a combination of biochemical and molecular methods. Among seven culturable bacteria isolated from the OS of SSB larvae, Enterobacter and Acinetobacter contributed to the suppression of JA signaling-related defenses in rice plants, and axenic larvae reinoculated with these two strains displayed better performance on rice plants. Our findings demonstrate that SSB larvae exploit oral secreted bacteria to interfere with plant anti-herbivore defense and avoid fully activating the JA-regulated antiherbivore defenses of rice plants. Key Message: The incorporation of antibiotics to field-collected Chilo suppressalis larvae significantly reduce bacterial quantities and affect larval performance on rice plants. Field-collected Chilo suppressalis larvae deposit abundant bacteria onto feeding sites through oral secretion, which modulate defense responses in rice plants. Bacteria isolated from larval oral secretion (such as Enterobacter and Acinetobacter) contribute to preventing rice plants to fully activate jasmonic acid-mediated antiherbivore defenses. Our results indicate insect-associated bacteria play an important role for C. suppressalis to adapt to rice plants, which could also be treated as targets for developing novel pest management strategies. [ABSTRACT FROM AUTHOR]

Ros/MucR is a widespread family of bacterial zinc-finger-containing proteins that integrate multiple functions, such as symbiosis, virulence, transcription regulation, motility, production of surface components, and various other physiological processes in cells. This regulatory protein family is conserved in bacteria and is characterized by its zinc-finger motif, which has been proposed as the ancestral domain from which the eukaryotic C2H2 zinc-finger structure has evolved. The first prokaryotic zinc-finger domain found in the transcription regulator Ros was identified in Agrobacterium tumefaciens. In the past decades, a large body of evidence revealed Ros/MucR as pleiotropic transcriptional regulators that mainly act as repressors through oligomerization and binding to AT-rich target promoters. The N-terminal domain and the zinc-finger-bearing C-terminal region of these regulatory proteins are engaged in oligomerization and DNA binding, respectively. These properties of the Ros/MucR proteins are similar to those of xenogeneic silencers, such as H-NS, MvaT, and Lsr2, which are mainly found in other lineages. In fact, a novel functional model recently proposed for this protein family suggests that they act as H-NS-'like' gene silencers. The prokaryotic zinc-finger domain exhibits interesting structural and functional features that are different from that of its eukaryotic counterpart (a βββα topology), as it folds in a significantly larger zinc-binding globular domain (a βββαα topology). Phylogenetic analysis of Ros/MucR homologs suggests an ancestral origin of this type of protein in α-Proteobacteria. Furthermore, multiple duplications and lateral gene transfer events contributing to the diversity and phyletic distribution of these regulatory proteins were found in bacterial genomes. [ABSTRACT FROM AUTHOR]

Background: The tomato leaf miner, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a major pest of tomato plants threatening global tomato production. The control of the pest is becoming increasingly difficult due to the rapid development of resistance to insecticides. Entomopathogenic nematodes (EPNs) of families Steinernematidae and Heterorhabditidae are successful biocontrol agents for many insect pests. Recently, their bacteria, Xenorhabdus spp. and Photorhabdus spp. have attracted great attention due to their major role in the pathogenicity of EPNs. In the present study, the pathogenicity of eleven EPNs isolates belonging to Steinernema feltiae and Heterorhabditis bacteriophora species was screened against the 1st/2nd and 3rd/4th instar larvae of T. absoluta. The cell-free supernatants and cell suspensions of the symbiotic bacteria from the most efficient isolates were further evaluated for their biocontrol potential in the oral and contact treatments on the larvae of T. absoluta. Results: KBC-4 and MCB-8 isolates of S. feltiae showed superior virulence relative to other EPNs species/isolates and induced 90% larval mortality against the 3rd/4th instar of T. absoluta larvae, whereas there were no clear differences in the efficacy of EPNs species/isolates against the 1st/2nd instar of T. absoluta. The 1st/2nd instar of T. absoluta larvae was more susceptible to cell-free supernatants and cell suspension of selected EPNs. The highest mortality (80%) was obtained from X. bovienii MCB-8 strain in the contact treatment of supernatants. In contact treatment of cell suspension, higher mortalities were obtained compared to oral treatments. Mortality rates ranged between 30 and 57.5% in the contact treatments of cell suspensions while the highest mortality did not exceed 20% in oral treatments. The antifeedant activity was observed in oral treatments of cell suspension and most of the larvae avoided feeding on treated leaves. Conclusion: The results indicated that symbiotic bacteria of EPNs had a great potential against T. absoluta larvae and contact treatment of cell-free supernatants against early instars of T. absoluta can be an ideal application. However, further studies are needed to investigate the field effectiveness of symbiotic bacteria. [ABSTRACT FROM AUTHOR]

A rhabditid entomopathogenic nematode (EPN), Oscheius chongmingensis, has a stable symbiotic relationship with the bacterial strain Serratia nematodiphila S1 harbored in its intestines and drastically reduced viability when associated with a non-native strain (186) of the same bacterial species. This nematode is thus a good model for understanding the molecular mechanisms and interactions involved between a nematode host and a member of its intestinal microbiome. Transcriptome analysis and RNA-seq data indicated that expression levels of the majority (8797, 87.59%) of mRNAs in the non-native combination of O. chongmingensis and S. nematodiphila 186 were downregulated compared with the native combination, including strain S1. Accordingly, 88.84% of the total uniq-sRNAs mapped in the O. chongmingensis transcriptome were specific between the two combinations. Six DEGs, including two transcription factors (oc-daf-16 and oc-goa-1) and four kinases (oc-pdk-1, oc-akt-1, oc-rtk, and oc-fak), as well as an up-regulated micro-RNA, oc-miR-71, were found to demonstrate the regulatory mechanisms underlying diminished host viability induced by a non-native bacterial strain. Oc-rtk and oc-fak play key roles in the viability regulation of O. chongmingensis by positively mediating the expression of oc-daf-16 to indirectly impact its longevity and stress tolerances and by negatively regulating the expression of oc-goa-1 to affect the olfactory chemotaxis and fecundity. In response to the stress of invasion by the non-native strain, the expression of oc-miR-71 in the non-native combination was upregulated to downregulate the expression of its targeting oc-pdk-1, which might improve the localization and activation of the transcription factor DAF-16 in the nucleus to induce longevity extension and stress resistance enhancement to some extent. Our findings provide novel insight into comprehension of how nematodes deal with the stress of encountering novel potential bacterial symbionts at the physiological and molecular genetic levels and contribute to improved understanding of host–symbiont relationships generally. [ABSTRACT FROM AUTHOR]

The coastal areas of the Red Sea within Saudi Arabia have a wide diversity of marine soft corals harboring many symbiotic bacteria that could present a potential source of many antibacterial metabolites. This study focused on the molecular characterization of antibiotic-producing bacteria associated with soft corals and investigated the influence of culture conditions on the production of their antibacterial extracts. Two soft coral samples were collected offshore from Yanbu City and identified as genus Sarcophyton. Screening for antibiotic-producing bacterial symbionts uncovered nine isolates with a wide inhibitory spectrum against gram-positive and negative bacteria. Phylogenetic analysis of the bioactive bacterial symbionts 16S rRNA gene sequences revealed the prevalence of gram-negative bacteria with seven isolates all belonging to class γ-proteobacteria. The remaining two isolates were represented within class firmicutes and actinobacteria. Eight isolates were grouped perfectly with their best matches. While the remaining ninth isolate was phylogenetically separated from all the species within the Photobacterium genus, suggesting that it could be a novel species. The optimum antibacterial compounds production was obtained in minimal media, at a temperature range of 25–28 °C, pH 7 and 100–150 rev/min agitation during 24–48 h of the incubation period, with moderate to high degree of thermal stability. [ABSTRACT FROM AUTHOR]

Introduction: Bacteria play critical roles in the reproduction, metabolism, physiology, and detoxification of their insect hosts. The ladybird beetle (Harmonia axyridis) harbors a myriad of endosymbiotic microbes. However, to date, little is known about how the microbial composition of H. axyridis varies throughout its life cycle. Methods: In this study, 16S rRNA amplicon sequencing and quantitative PCR were employed to investigate the diversity and dynamics of bacterial symbionts across the egg, larval, pupae, and adults stages of H. axyridis. Results: Higher bacterial community richness and diversity were observed in eggs, followed by those in adults and pupae. The community richness index differed significantly between second-instar larvae and other developmental stages. Proteobacteria, Firmicutes, and Actinobacteria were the dominant phyla. Staphylococcus, Enterobacter, Glutamicibacter, and Acinetobacter were the dominant bacteria genera; however, their relative abundances fluctuated across host developmental stages. Interestingly, the larval stage harbored high proportions of Firmicutes, whereas the adult microbial community largely consisted of Proteobacteria. Discussion: This study is the first to determine the symbiotic bacterial composition across key life stages of H. axyridis. These outcomes can foster the development of environmental risk assessments and novel biological control strategies. [ABSTRACT FROM AUTHOR]

Microalgae-bacteria consortia (MBC) system has been shown to enhance the efficiency of microalgae in wastewater treatment, yet its effectiveness in treating levofloxacin (LEV) wastewater remains unexplored. This study compared the treatment of LEV wastewater using pure Chlorella pyrenoidosa (PA) and its MBC constructed with activated sludge bacteria. The results showed that MBC improved the removal efficiency of LEV from 3.50–5.41 % to 33.62–57.20 % by enhancing the growth metabolism of microalgae. The MBC increased microalgae biomass and extracellular polymeric substance (EPS) secretion, yet reduced photosynthetic pigment content compared to the PA. At the phylum level, Proteobacteria and Actinobacteriota are the major bacteria in MBC. Furthermore, the transcriptome reveals that the growth-promoting effects of MBC are associated with the up-regulation of genes encoding the glycolysis, the citrate cycle (TCA cycle), and the pentose phosphate pathway. Enhanced carbon fixation, coupled with down-regulation of photosynthetic electron transfer processes, suggests an energy allocation mechanism within MBC. The up-regulation of porphyrin and arachidonic acid metabolism, along with the expression of genes encoding LEV-degrading enzymes, provides evidence of MBC's superior tolerance to and degradation of LEV. Overall, these findings lead to a better understanding of the underlying mechanisms through which MBC outperforms PA in treating LEV wastewater. [Display omitted] • Microalgae-bacteria consortia (MBC) is effective at removing levofloxacin (LEV). • Decarboxylation and defluorination were the main LEV degradation mechanisms. • MBC and LEV promote algal biomass and extracellular polymeric substance secretion. • Proteobacteria and Actinobacteriota are the major bacteria in MBC. • MBC upregulates genes encoding enzymes for energy metabolism and LEV degradation. [ABSTRACT FROM AUTHOR]

[Display omitted] • EPN symbiotic bacteria and their filtrates have insecticidal effects on T. absoluta. • The bacterium X. nematophila was the most effective against larvae of T. absoluta. • 1st/2nd instar larvae of T. absoluta were more susceptible than 3rd/4th instar. • The EPN, their bacteria and filtrates were also tested on the key predator N. tenuis. • The EPN symbiotic bacteria and their filtrates had low impact on N. tenuis. The use of biocontrol agents, such as predators and entomopathogenic nematodes, is a promising approach for the effective control of the tomato leafminer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidaean), an oligophagous insect feeding mainly on Solanaceae species and a major pest of field- and greenhouse-grown tomatoes globally. In this context, the effects of two entomopathogenic nematode species Steinernema carpocapsae (Weiser) (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora (Poinar) (Rhabditida: Heterorhabditidae), as well as their respective bacterial symbionts, Xenorhabdus nematophila and Photorhabdus luminescens (Enterobacterales: Morganelaceae), which were applied as bacterial cell suspensions and as crude cell-free liquid filtrates on T. absoluta larvae, were investigated. The results showed that of all treatments, the nematodes S. carpocapsae and H. bacteriophora were the most effective, causing up to 98 % mortality of T. absoluta larvae. Regarding bacteria and their filtrates, the bacterium X. nematophila was the most effective (69 % mortality in young larvae), while P. luminescens and both bacterial filtrates showed similar potency (ca. 48–55 % mortality in young larvae). To achieve a holistic approach of controlling this important pest, the impact of these factors on the beneficial predator Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae) was also studied. The results demonstrated that although nematodes and especially S. carpocapsae , caused significant mortality on N. tenuis (87 %), the bacterial cell suspensions of X. nematophila and P. luminescens and crude cell-free liquid filtrates had minimum impact on this beneficial predator (∼11–30 % mortality). [ABSTRACT FROM AUTHOR]

The competitiveness of Bradyrhizobium japonicum inoculation strain against indigenous rhizobia was examined in a soil pot experiment. The effect of inoculation strain was evaluated under different soil conditions: with or without previously grown soybean and applied commercial inoculant. Molecular identification of inoculation strain and investigated rhizobial isolates, obtained from nodules representing inoculated treatments, was performed based on 16S rDNA and enterobacterial repetitive intergenic consensus (ERIC) sequencing. Inoculation strain showed a significant effect on the investigated parameters in both soils. Higher nodule occupancy (45% vs. 18%), nodule number (111% vs. 5%), nodule dry weight (49% vs. 9%), shoot length (15% vs. 7%), root length (31% vs. 13%), shoot dry weight (34% vs. 11%), shoot nitrogen content (27% vs. 2%), and nodule nitrogen content (9% vs. 5%) was detected in soil without previously grown soybean and applied commercial inoculant. Soil had a significant effect on the shoot, root and nodule nitrogen content, while interaction of experimental factors significantly altered dry weight and nitrogen content of shoots, roots and nodules, as well as number of nodules. Nodulation parameters were significantly related with shoot dry weight, shoot and nodule nitrogen content. Symbiotic performance of inoculation strains in the field could be improved through co-selection for their competitiveness and effectiveness.

The wheat aphid, Sitobion miscanthi, is one of the most destructive pests of wheat plants in the temperate regions of China. Little is known about the genetic structure evolution of the different geographic populations of S. miscanthi with its migration. In this study, we investigated the population genetic structure and demographic history of S. miscanthi by analysing 18 geographical populations across China using one mitochondrial gene, COI; one nuclear gene, EF-1α; and two endosymbiont Buchnera genes, gnd and trpA. Analysis of data from the various groups showed high haplotype diversity and low nucleotide variation. SAMOVA analysis did not find a correlation between genetic distance and geographic distance. However, areas with high population diversity exhibited high haplotype diversity. Therefore, we speculate that there are two main natural migration pathways of S. miscanthi in China. One is from Yunnan to the Sichuan Basin, and the other is from Wuhan, Xinyang and Jiaodong Peninsula areas to the northwest. Based on this hypothesis, we inferred that these aphid populations appear first in the southwestern and southern regions and spread to the north with the help of the southeastern and southwestern monsoons, which occur in spring and summer. In autumn, the aphids spread southward with the northeastern and northwestern monsoons.

ABSTRACT Many insects contain endosymbiotic bacteria within their bodies. In multiple endosymbiotic systems comprising two or more symbionts, each of the symbionts is generally localized in a different host cell or tissue. Bemisia tabaci (Sweet potato whitefly) possesses a unique endosymbiotic system where co-obligate symbionts are localized in the same bacteriocytes. Using fluorescence in situ hybridization, we found that endosymbionts in B. tabaci MEAM1 occupy distinct subcellular habitats, or niches, within a single bacteriocyte. Hamiltonella was located adjacent to the nucleus of the bacteriocyte, while Portiera was present in the cytoplasm surrounding Hamiltonella. Immunohistochemical analysis revealed that the endoplasmic reticulum separates the two symbionts. Habitat segregation was maintained for longer durations in female bacteriocytes. The same segregation was observed in three genetically distinct B. tabaci groups (MEAM1, MED Q1, and Asia II 6) and Trialeurodes vaporariorum, which shared a common ancestor with Bemisia over 80 million years ago, even though the coexisting symbionts and the size of bacteriocytes were different. These results suggest that the habitat segregation system existed in the common ancestor and was conserved in both lineages, despite different bacterial partners coexisting with Portiera. Our findings provide insights into the evolution and maintenance of complex endosymbiotic systems and highlight the importance of organelles for the construction of separate niches for endosymbionts. IMPORTANCE Co-obligate endosymbionts in B. tabaci are exceptionally localized within the same bacteriocyte (a specialized cell for endosymbiosis), but the underlying mechanism for their coexistence remains largely unknown. This study provides evidence for niche segregation at the subcellular level between the two symbionts. We showed that the endoplasmic reticulum is a physical barrier separating the two species. Despite differences in co-obligate partners, this subcellular niche segregation was conserved across various whitefly species. The physical proximity of symbionts may enable the efficient biosynthesis of essential nutrients via shared metabolic pathways. The expression “Good fences make good neighbors” appears to be true for insect endosymbiotic systems.

ABSTRACT Gut symbiotic bacteria are known to be closely related to insect development, nutrient metabolism, and disease resistance traits, but the most important factors leading to changes in these communities have not been well clarified. To address this, we examined the associations between the gut symbiotic bacteria and the host genotype and geographical distribution of Solenopsis invicta in China, where it is invasive and has spread primarily by human-mediated dispersal. Thirty-two phyla were detected in the gut symbiotic bacteria of S. invicta. Proteobacteria were the most dominant group among the gut symbiotic bacteria. Furthermore, the Bray-Curtis dissimilarity matrices of the gut symbiotic bacteria were significantly positively correlated with the geographical distance between the host ant colonies, but this relationship was affected by the social form. The distance between monogyne colonies had a significant effect on the Bray-Curtis dissimilarity matrices of gut symbiotic bacteria, but the distance between polygyne colonies did not. Moreover, the Bray-Curtis dissimilarity matrices were positively correlated with Nei’s genetic distance of the host but were not correlated with the COI-based genetic distance. This study provides a scientific basis for further understanding the ecological adaptability of red imported fire ants during invasion and dispersal. IMPORTANCE We demonstrated that gut microbiota composition and diversity varied among populations. These among-population differences were associated with host genotype and geographical distribution. Our results suggested that population-level differences in S. invicta gut microbiota may depend more on environmental factors than on host genotype.

This article deals with the external and internal morphology in the adult stages of Nereilinum murmanicum Ivanov, 1961, a widespread species of Siboglinidae in the Barents Sea. Since Siboglinidae is currently a taxon within the phylum Annelida, we revised the commonly held view of the body segmentation pattern typical of the species and provided the first des­cription of its opisthosoma. Furthermore, the postannular region and trophosome were structurally analyzed using histology and electron microscopy. The endodermal origin of the trophosome was suggested from its structure and position. The juvenile and larval stages of the species were described. The early larvae were found to resemble the trochophores in Annelida. The late larvae look like the late metatrochophora in Annelida and lack parapodia. The juvenile specimens retain no larval traits and are morphologically comparable to the adult stages. New data on the ecology of the species were obtained: it appears to also inhabit deeper water layers. The boundaries of its range in the Barents Sea were expanded and specified. The abundance distribution of the species in the Barents Sea (30–40 ind/m2 for the major part of the sea, up to 72–113 ind/ m2 for certain areas of the sea in particular years) was considered.

Symbiotic bacteria on animal hosts can prevent pathogenic bacterial infections by several mechanisms. Among them, symbiotic bacteria can indirectly enhance host's immune responses or, directly, produce antimicrobial substances against pathogens. Due to differences in life-style, different host species are under different risks of microbial infections. Consequently, if symbiotic bacteria are somewhat selected by genetically determined host characteristics, we would expect the antimicrobial properties of bacterial symbionts to vary among host species and to be distributed according to risk of infection. Here we have tested this hypothesis by measuring the antimicrobial ability of the bacterial strains isolated from the uropygial-gland skin of 19 bird species differing in nesting habits, and, therefore, in risk of microbial infection. In accordance with our predictions, intensity and range of antimicrobial effects against the indicator strains assayed varied among bird species, with hole-and open-nesters showing the highest and the lowest values, respectively. Since it is broadly accepted that hole-nesters have higher risks of microbial infection than open nesters, our results suggest that the risk of infection is a strong driver of natural selection to enhance immunocompetence of animals through selecting for antibiotic-producing symbionts. Future research should focus on characterizing symbiotic bacterial communities and detecting coevolutionary processes with particular antibiotic-producing bacteria within-host species. [ABSTRACT FROM AUTHOR]

Purpose: The root-associated microbiota is essential to plant health, fitness and productivity, but the effect of plant domestication on the ecological process of microbial community assembly is unclear. Methods: High-throughput sequencing of the 16S rRNA genes was employed to investigated the the diversity, ecological assembly process and cooccurrence relationship of the bacterial communities of multiple root compartment niches (root zone, rhizosphere, and root endosphere) between wild and landrace accessions grown in three soil types. Results: Our results showed that the domestication effect on bacterial community increased from the root zone to the rhizosphere and endosphere, while the soil type effect decreased. Compared with wild soybean, the root endosphere bacterial community of the landraces was more sensitive to soil environmental change. The deterministic process dominated the assembly of the bacterial community in the rhizosphere and root endosphere, and its relative contribution was higher in the landraces than in wild soybeans. In the two root compartments, the increased root system selection pressure in the landraces was indicated by a greater loss of bacteria at the same taxon level and lower bacterial diversity. Furthermore, the family Oxalobacteraceae and the class Actinobacteria were identified as important root-associated biomarker taxa for wild soybeans, while Enterobacteriaceae was such for the landraces. Conclusions: Our findings provide crucial empirical evidence for the host selection and enrichment process of the microbial community under local domestication and are of great significance in understanding the coevolution of hosts and microbiota, which will aid in manipulating microbiota for future crop breeding. [ABSTRACT FROM AUTHOR]

The soil-dwelling nematode Steinernema feltiae is found across a wide range of environmental conditions. We asked if its only bacterial symbiont, Xenorhabdus bovienii, shows intraspecific variability in its thermal range, which may affect effectiveness of S. feltiae against host insects. We isolated X. bovienii from S. feltiae from six different natural locations with different mean annual temperatures and two laboratory cultures. We estimated X. bovienii thermal range and determined the specific growth rate based on optical density measurements and mathematical modeling using the Ratkowsky model. The minimal temperature (Tmin) of X. bovienii growth ranged from 0.9 ± 2.2 °C to 7.1 ± 1.4 °C. The optimal temperature (Topt) varied between 25.1 ± 0.2 °C and 30.5 ± 0.2 °C. The model showed that X. bovienii stops multiplying at around 36 °C. The calculated specific X. bovienii growth rate ranged from 2.0 ± 0.3 [h−1] to 3.6 ± 0.5 [h−1]. No differences in Tmin, Topt, and Tmax between the isolated bacteria were found. Additionally, X. bovieniiTopt did not correlate with the mean annual temperature of S. feltiae origin. However, the obtained growth curves suggested that the analyzed X. bovienii may show some variability when comparing the growth curves characteristics. [ABSTRACT FROM AUTHOR]

The fine structure of the body wall and gut was for the first time studied in the competent larvae of the frenulate pogonophoran Siboglinum fiordicum. Mass apoptosis of cell nuclei was observed in the dermo-muscular body wall and coelomic epithelium. Apoptotic nuclei were found in both cell cytoplasm and outside of the larval body. In the latter case, each nucleus was surrounded by the plasmalemma, and the entire cluster was covered with the cuticle. Cells of the larval gut retained the usual structure with the cytoplasm filled with numerous yolky granules and the nucleus displaying usual morphology. Similar apoptotic processes have been described in vestimentiferans and found to be initiated by penetration of symbiotic bacteria through the integument into the dorsal mesentery. The process of apoptotic rearrangement of body wall cells and the formation of unique symbiosis with bacteria were assumed to be time-spaced in S. fiordicum, occurring sequentially rather than simultaneously, unlike in vestimentiferans. [ABSTRACT FROM AUTHOR]

BACKGROUND Colorado potato beetle (CPB; Leptinotarsa decemlineata) is a destructive quarantine pest that develops broad physiological adaptations to potato plants. During feeding, CPB deposits a copious amount of wet frass onto the surface of leaves and stems that remains in place for long periods. Insect behaviors such as feeding, crawling and oviposition are able to mediate plant defenses. However, the specific role of CPB defecation‐associated cues in manipulating plant defenses remains unclear. RESULTS: CPB larval frass significantly suppressed potato polyphenol oxidase activity and enhanced larval growth on treated potato plants. The incorporation of antibiotics into larval frass triggered higher jasmonic acid (JA)‐regulated defense responses in potato plants compared with antibiotic‐free frass. Four bacterial symbionts belonging to the genera Acinetobacter, Citrobacter, Enterobacter and Pantoea were isolated from larval frass and suppressed plant defenses. After reinoculation of these bacteria into axenic larvae, Acinetobacter and Citrobacter were found to be highly abundant in the frass, whereas Enterobacter and Pantoea were less abundant probably due to the negative effect of potato steroidal glycoalkaloids (SGA) such as α‐solanine. Furthermore, direct application of Acinetobacter and Citrobacter to wounded potato plants significantly inhibited the expression of genes associated with the JA‐mediated defense signaling pathway and SGA biosynthesis. CONCLUSION: Our findings demonstrate that CPB exploits frass‐associated bacteria as a deceptive strategy of plant defense suppression, adding an interesting dimension to our understanding of how CPB successfully specializes on potato plants. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Endosymbiotic bacteria significantly impact the fitness of their arthropod hosts. Dermanyssus gallinae , the poultry red mite, is a blood-feeding ectoparasite that exclusively feeds on avian blood. While there is a relatively comprehensive understanding of its microbial community structures across developmental stages based on 16S rRNA sequencing， the functional integration of these microbes within the host's physiology remains elusive. This study aims to elucidate the role of symbiotic bacteria in D. gallinae biology. 16S rRNA amplicon sequencing and fluorescence in situ hybridization revealed a prominent midgut-confinement bacterial microbiota with considerable diversity, out of which Kocuria and Bartonella A acted as the predominant bacterial genera inhabiting D. gallinae . The relative abundance of Bartonella A increased rapidly after blood-sucking, suggesting its adaptation to a blood-based diet and its pivotal role in post-engorgement activities. Some of the isolated bacterial strains from D. gallinae display hemolytic activity on blood agar, potentially aiding blood digestion. To corroborate this in vivo , antibiotic-mediated clearance was exploited to generate dysbiosed cohorts of D. gallinae mites, lacking some of the key bacterial species. Phenotypic assessments revealed that dysbiosed mites experienced delayed blood digestion and diminished reproductive capacity. Whole-genome sequencing identified Bartonella A as a new species within the genus Bartonella , exhibiting characteristics of an obligate symbiont. These findings underscore the significance of microbiota in poultry red mites and suggest microbiota-targeted strategies for controlling mite populations in poultry farms., Competing Interests: None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of the International Society for Microbial Ecology.)

The microbiota plays a crucial role in inducing immune responses. Our previous studies have shown that symbiotic bacterial sensing by the nucleotide-binding oligomerization-domain-containing protein 2 (Nod2) receptor is involved in the mucosal adjuvanticity of cholera toxin. However, cholera toxin's potential toxicity limits human use. Here, screening other less toxic adjuvants showed that toll-like receptor (TLR) 4 and 7 agonists synergized with the microbiota in inducing adaptive immune responses upon nasal immunization. Particularly, Imiquimod, a TLR7 agonist, exhibited synergistic effects with bacterial component MDP, a Nod2 ligand, in inducing immune responses, such as IL-12p40 and IL-6 productions in bone marrow-derived dendritic cells (BMDCs) and follicular helper T (T FH ) cell differentiation and high-affinity antibody production in immunized mice. The Imiquimod-MDP combination notably elicited immune protection against influenza and SARS-CoV-2 infections. Furthermore, we isolated some bacteria from the nasal cavity of healthy donors, and their Nod2-stimulating activities were measured using a reporter cell line. Staphylococcus aureus, with notable Nod2-stimulating activity, showed higher synergy with Imiquimod than Staphylococcus epidermidis, while the synergistic effects by Imiquimod-bacteria combination disappeared in Nod2-knockout mice. Moreover, the pretreatment with S. aureus enhanced the protective effect of Imiquimod-mediated vaccination against influenza virus compared to S. epidermidis. These results imply that the Imiquimod-MDP and the Imiquimod-bacteria combinations could be novel and promising complex adjuvants in developing intranasal vaccines., Competing Interests: Competing Interests None., (Copyright © 2024. Published by Elsevier Ltd.)

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.IMPORTANCEDiverse 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., Competing Interests: The authors declare no conflict of interest.

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., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhang, Chen, Zeng, Deng, Liu, Li, Hou, Zhou, Bao, Hou, Weng, He and Huang.)

A cDNA encoding a phage-type lysozyme, designated as HcPLYZ, was successfully cloned from Hyriopsis cumingii. The full-length cDNA sequence of HcPLYZ was determined to be 896 base pairs in length. Analysis revealed the absence of a signal peptide at its N-terminus, and identified two highly conserved phage-type lysozyme activity sites, Glu 20 and Asp 29 , within the deduced amino acid sequence of HcPLYZ. The results of the cloning and sequencing symbiotic bacteria in tissues were consistent with those obtained using tissue cDNA as the template, suggesting that HcPLYZ may originate a symbiotic bacterium. The expression levels of HcPLYZ mRNA exhibited significant variations across different tissues. Successful expression was induced using IPTG, and the native recombinant protein was subsequently purified through affinity chromatography employing Ni 2+ , and the optimal pH and temperature of which were determined to be 5.5 and 50 °C, respectively. Following exposure to Aeromonas hydrophila, there was a significant increase in the levels of HcPLYZ mRNA in the hemocytes, hepatopancreas, and gills. HcPLYZ was demonstrated the inhibition activity of 55% and 83% against Micrococcus lysodeikticus under pH 5.5 and 50 °C conditions, respectively. These results suggested that HcPLYZ possessed antibacterial activity against both A. hydrophila and M. lysodeikticus., (Copyright © 2024. Published by Elsevier Ltd.)

Abstract Complex interactions between symbiotic bacteria and insects ultimately result in equilibrium in all aspects of life in natural insect populations. In this study, abundance of principal symbiotic bacteria was estimated using qPCR in 1553 individuals of aphids, Aphis gossypii. Aphids were sampled from primary and secondary host plants—hibiscus and cotton. Hibiscus aphids were collected from 24 different locations in April, September, and November, whereas cotton aphids were collected between 2015 and 2017 from areas with wide variations in climatic conditions. About 30%–45% aphids were recorded with the most dominant symbiont, Arsenophonus. The other symbionts were in low frequency, and about 7% of aphids were noted with Hamiltonella, Acinetobacter, and Microbacterium, and 3% of aphids were verified with Serratia and Pseudomonas. Aphids infected with Hamiltonella, Arsenophonus, and Serratia can influence Buchnera densities. Hamiltonella has positive interaction with densities of Arsenophonus and Serratia. Almost 100% coinfection of Hamiltonella and Arsenophonus was detected in Xinxiang aphids and 50% coinfection was reported in aphids from North China, while no coinfection was detected in Hainan aphids. These findings describe the prevalence pattern and richness of core community of symbiotic bacteria in naturally occurring populations of A. gossypii and provide new insights for the study of symbiotic bacteria.

Symbiotic bacteria on animal hosts can prevent pathogenic bacterial infections by several mechanisms. Among them, symbiotic bacteria can indirectly enhance host’s immune responses or, directly, produce antimicrobial substances against pathogens. Due to differences in life-style, different host species are under different risks of microbial infections. Consequently, if symbiotic bacteria are somewhat selected by genetically determined host characteristics, we would expect the antimicrobial properties of bacterial symbionts to vary among host species and to be distributed according to risk of infection. Here we have tested this hypothesis by measuring the antimicrobial ability of the bacterial strains isolated from the uropygial-gland skin of 19 bird species differing in nesting habits, and, therefore, in risk of microbial infection. In accordance with our predictions, intensity and range of antimicrobial effects against the indicator strains assayed varied among bird species, with hole-and open-nesters showing the highest and the lowest values, respectively. Since it is broadly accepted that hole-nesters have higher risks of microbial infection than open nesters, our results suggest that the risk of infection is a strong driver of natural selection to enhance immunocompetence of animals through selecting for antibiotic-producing symbionts. Future research should focus on characterizing symbiotic bacterial communities and detecting coevolutionary processes with particular antibiotic-producing bacteria within-host species.

Sponge-associated bacteria are a well-known source of unique and diverse natural products with potential biological activities. In this study, a rarely occurring sponge-associated bacterium from the genus Alcanivorax was isolated from the sponge Siphonochalina siphonella and subjected to extraction of metabolites. Metabolites were extracted from the culture supernatant and biomass of the bacterium using the solvent extraction method. Extracts were evaluated for antibacterial and antibiofilm activities against five different biofilm-forming bacteria isolated from the microfouling assemblage. The crude extracts were subjected to chemical profiling to identify their composition using gas chromatography-coupled mass spectrometry (GC-MS). The results obtained show mild to strong antibacterial activity of the extracts against the biofilm-forming bacteria with the minimum inhibitory concentration (MIC) of 3.125 mg ml-1. The percentage inhibition of biofilm formation ranged from 46% to 71%. Compounds detected in GC-MS analysis are mostly fatty acids and their derivatives that include compounds with known antibiofilm activity such as tetradecanoic acid, dodecanoic acid, and hexadecanol from the culture supernatant extract. Eicosane; hexadecanoic acid, methyl ester; and hexadecanoic acid, ethyl ester were detected from the biomass extract. Molecular docking analysis of the compounds from the bacterial extracts confirms their potential antibiofilm properties. The result indicated that Alcanivorax sp. associated with the sponge possess relevant antibiofilm activity and may serve as a novel source of secondary metabolites with the application as antibiofilm agents.

The population divergence process of deep-sea vent invertebrates is driven by both biotic (e.g., dispersal during the larval stage) and abiotic factors such as deep-ocean currents, depth, and the geological setting of vents. However, little is known regarding the divergence of hydrothermal vent microorganisms. Therefore, our study sought to investigate the influence of geological and geographic factors on the divergence of symbiotic bacteria of Bathymodiolus vent mussels. The genetic differentiation patterns of symbionts were examined using next-generation sequencing DNA data in two ocean basins with distinct geological features: the slow-spreading Central Indian Ridge (CIR) and the fast- or superfast-spreading eastern Pacific Ridges. Our findings showed that the degree of differentiation of symbiont populations was geographically hierarchical: the highest between ocean basins, followed by inter-ridge sites between the East Pacific Rise and the Pacific Antarctic Ridge. The Easter Microplate intervening these two ridges acted as a biogeographic physical barrier for both symbionts and their host mussels. On a scale of intra-ridge, symbionts showed isolation by distance in the CIR but not in the eastern Pacific ridges. These contrasting genetic patterns relate to different ridge spreading rates determining most of the geological characteristics of mid-ocean ridges that affect the connectivity of vent habitats in space and time. At the intra-ridge geographic scale of the CIR, population divergence processes of both symbionts and hosts from separate three ridge segments were analyzed in detail using a genetic model of isolation with migration (IM). The phylogenetic topology of symbiont populations was congruent with the host populations, indicating the influence of common historical and physical constraints for habitats and dispersal between vents in the Central Indian Ridge. Collectively, our findings provide key insights into the dynamics of microbial population divergence in deep-sea vents.

Rhipicephalus sanguineus, the brown dog tick, is the most widespread tick in the world and a predominant vector of multiple pathogens affecting wild and domestic animals. There is an increasing interest in understanding the role of tick microbiome in pathogen acquisition and transmission as well as in environment–vector interfaces. Several studies suggested that the tick microbial communities are under the influence of several factors including the tick species, dietary bloodmeal, and physiological stress. Compared with insects, very little of the microbial community is known to contribute to the nutrition of the host. Therefore, it is of significance to elucidate the regulation of the microbial community of Rh. Sanguineus under starvation stress. Starvation stress was induced in wild-type adults (1 month, 2 months, 4 months, 6 months) and the microbial composition and diversity were analyzed before and after blood feeding. After the evaluation, it was found that the microbial community composition of Rh. sanguineus changed significantly with starvation stress. The dominant symbiotic bacteria Coxiella spp. of Rh. sanguineus gradually decreased with the prolongation of starvation stress. We also demonstrated that the starvation tolerance of Rh. sanguineus was as long as 6 months. Next, Coxiella-like endosymbionts were quantitatively analyzed by fluorescence quantitative PCR. We found a pronounced tissue tropism in the Malpighian tubule and female gonad, and less in the midgut and salivary gland organs. Finally, the blood-fed nymphs were injected with ofloxacin within 24 h. The nymphs were allowed to develop into adults. It was found that the adult blood-sucking rate, adult weight after blood meal, fecundity (egg hatching rate), and feeding period of the newly hatched larvae were all affected to varying degrees, indicating that the removal of most symbiotic bacteria had an irreversible effect on it.

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.

Symbiont mediated RNAi (SMR) is a promising method for precision control of pest insect species such as Western Flower Thrips (WFT). Two species of bacteria are known to be dominant symbiotic bacteria in WFT, namely BFo1 and BFo2 (Bacteria from Frankliniella occidentalis 1 and 2), as we here confirm by analysis of next-generation sequence data derived to obtain a reference WFT genome sequence. Our first demonstration of SMR in WFT used BFo2, related to Pantoea, isolated from a domesticated Dutch thrips population. However, for successful use of SMR as a thrips control measure, these bacteria need to successfully colonize different environmental thrips populations. Here, we describe a United Kingdom thrips population that does not harbour BFo2, but does contain BFo1, a species related to Erwinia. Attempts to introduce BFo2 indicate that this bacterium is unable to establish itself in the United Kingdom thrips, in contrast to successful colonization by a strain of BFo1 expressing green fluorescent protein. Fluorescence microscopy indicates that BFo1 occupies similar regions of the thrips posterior midgut and hindgut as BFo2. Bacterial competition assays revealed that a barrier to BFo2 establishing itself in thrips is the identity of the resident BFo1; BFo1 isolated from the United Kingdom thrips suppresses growth of BFo2 to a greater extent than BFo1 from the Dutch thrips that is permissive for BFo2 colonization. The ability of the latter strain of BFo1 to colonize the United Kingdom thrips is also likely attributable to its ability to out-compete the resident BFo1. Lastly, we observed that United Kingdom thrips pre-exposed to the Dutch BFo1 could then be successfully colonized by BFo2. These results indicate, for the first time, that microbial competition and strain differences can have a large influence on how symbiotic bacteria can colonize different populations of an insect species. [ABSTRACT FROM AUTHOR]

Symbiont mediated RNAi (SMR) is a promising method for precision control of pest insect species such as Western Flower Thrips (WFT). Two species of bacteria are known to be dominant symbiotic bacteria in WFT, namely BFo1 and BFo2 (Bacteria from Frankliniella occidentalis 1 and 2), as we here confirm by analysis of next-generation sequence data derived to obtain a reference WFT genome sequence. Our first demonstration of SMR in WFT used BFo2, related to Pantoea, isolated from a domesticated Dutch thrips population. However, for successful use of SMR as a thrips control measure, these bacteria need to successfully colonize different environmental thrips populations. Here, we describe a United Kingdom thrips population that does not harbour BFo2, but does contain BFo1, a species related to Erwinia. Attempts to introduce BFo2 indicate that this bacterium is unable to establish itself in the United Kingdom thrips, in contrast to successful colonization by a strain of BFo1 expressing green fluorescent protein. Fluorescence microscopy indicates that BFo1 occupies similar regions of the thrips posterior midgut and hindgut as BFo2. Bacterial competition assays revealed that a barrier to BFo2 establishing itself in thrips is the identity of the resident BFo1; BFo1 isolated from the United Kingdom thrips suppresses growth of BFo2 to a greater extent than BFo1 from the Dutch thrips that is permissive for BFo2 colonization. The ability of the latter strain of BFo1 to colonize the United Kingdom thrips is also likely attributable to its ability to out-compete the resident BFo1. Lastly, we observed that United Kingdom thrips pre-exposed to the Dutch BFo1 could then be successfully colonized by BFo2. These results indicate, for the first time, that microbial competition and strain differences can have a large influence on how symbiotic bacteria can colonize different populations of an insect species.

• H 2 O 2 induction led to the release of catalase KatA1 from the cells of P. aquatica HC52. • H 2 O 2 conditions induced OMV production in P. aquatica HC52. • KatA1 was identified within the total secreted proteins and OMVs of P. aquatica HC52. • Extracellular catalase KatA1 could protect M. aeruginosa PCC7806 from H 2 O 2 exposure. High levels of environmental H 2 O 2 represent a threat to many freshwater bacterial species, including toxic-bloom-forming Microcystis aeruginosa , particularly under high-intensity light conditions. The highest extracellular catalase activity-possessing Pseudoduganella aquatica HC52 was chosen among 36 culturable symbiotic isolates from the phycosphere in freshly collected M. aeruginosa cells. A zymogram for catalase activity revealed the presence of only one extracellular catalase despite the four putative catalase genes (katA1, katA2, katE , and srpA) identified in the newly sequenced genome (∼6.8 Mb) of P. aquatica HC52. Analysis of secreted catalase using liquid chromatography–tandem mass spectrometry was identified as KatA1, which lacks a typical signal peptide, although the underlying mechanism for its secretion is unknown. The expression of secreted KatA1 appeared to be induced in the presence of H 2 O 2. Proteomic analysis also confirmed the presence of KatA1 inside the outer membrane vesicles secreted by P. aquatica HC52 following exposure to H 2 O 2. High light intensities (> 100 µmol m−2 s−1) are known to kill catalase-less axenic M. aeruginosa cells , but the present study found that the presence of P. aquatica cells supported the growth of M. aeruginosa , while the extracellular catalases in supernatant or purified form also sustained the growth of M. aeruginosa under the same conditions. Our results suggest that the extracellular catalase secreted by P. aquatica HC52 enhances the tolerance of M. aeruginosa to H 2 O 2 , thus promoting the formation of M. aeruginosa blooms under high light intensities. [Display omitted] [ABSTRACT FROM AUTHOR]

The rhizosphere system of plants hosts a diverse consortium of bacteria that confer beneficial effects on plant, such as plant growth-promoting rhizobacteria (PGPR), biocontrol agents with disease-suppression activities, and symbiotic nitrogen fixing bacteria with the formation of root nodule. Efficient colonization in planta is of fundamental importance for promoting of these beneficial activities. However, the process of root colonization is complex, consisting of multiple stages, including chemotaxis, adhesion, aggregation, and biofilm formation. The secondary messenger, c-di-GMP (cyclic bis-(3′-5′) dimeric guanosine monophosphate), plays a key regulatory role in a variety of physiological processes. This paper reviews recent progress on the actions of c-di-GMP in plant beneficial bacteria, with a specific focus on its role in chemotaxis, biofilm formation, and nodulation. [ABSTRACT FROM AUTHOR]

Organic carbon source plays a fundamental function in the metabolism and production of photosynthetic microorganisms. Herein, this study explored the effects of carbon source additions (glucose, sucrose and sodium acetate) on growth and biochemical components of the economic cyanobacterium Nostoc sphaeroides. The effects of sucrose and sodium acetate on its metabolomics changes, as well as the diversity and community composition of symbiotic bacteria, were analyzed. Results showed that sucrose noticeably enhanced the relative growth rate of N. sphaeroides. The content of phycocyanin was significantly greater under sodium acetate than under other carbon sources. Some metabolites associated with amino acid biosynthesis was significantly enriched under sodium acetate, implying its critical role in phycocyanin synthesis. Sodium acetate had a significant impact on the diversity and community composition of symbiotic bacteria. Additionally, partial symbiotic bacteria were shown to be involved in the amino acid metabolism of N. sphaeroides. These findings provided valuable insights into the mechanisms of N. sphaeroides on its metabolic responses to carbon source changes and the synergy of its symbiotic bacteria, contributing to the establishment of effective carbon source supply strategies for artificial cultivation and resource restoration. • Organic carbon is critical compound for the growth and metabolism of N. sphaeroides. • Organic carbon visibly affected symbiotic bacteria diversity and composition. • The biomass of N. sphaeroides was markedly enhanced by sucrose addition. • Acetate sodium significantly promoted the accumulation of phycocyanin. • Symbiotic bacteria were likely to be involved in the metabolism of amino acids. [ABSTRACT FROM AUTHOR]

Symbiotic bacteria and hormesis in aphids are the driving forces for pesticide resistance. However, the mechanism remains unclear. In this study, the effects of imidacloprid on the population growth parameters and symbiotic bacterial communities of three successive generations of Acyrthosiphon gossypii were investigated. The bioassay results showed that imidacloprid had high toxicity to A. gossypii with an LC50 of 1.46 mg·L−1. The fecundity and longevity of the G0 generation of A. gossypii decreased when exposed to the LC15 of imidacloprid. The net reproductive rate (R0), intrinsic rate of increase (rm), finite rate of increase (λ), and total reproductive rate (GRR) of G1 and G2 offspring were significantly increased, but those of the control and G3 offspring were not. In addition, sequencing data showed that the symbiotic bacteria of A. gossypii mainly belonged to Proteobacteria, with a relative abundance of 98.68%. The dominant genera of the symbiotic bacterial community were Buchnera and Arsenophonus. After treatment with the LC15 of imidacloprid, the diversity and species number of bacterial communities of A. gossypii decreased for G1–G3 and the abundance of Candidatus-Hamiltonella decreased, but Buchnera increased. These results provide insight into the resistance mechanism of insecticides and the stress adaptation between symbiotic bacteria and aphids.

Symbiotic bacteria form a mutualistic relationship with nematodes and are pathogenic to many insect pests. They kill insects using various strategies to evade or suppress their humoral and cellular immunity. Here we evaluate the toxic effects of these bacteria and their secondary metabolites on the survival and phenoloxidase (PO) activation of Octodonta nipae larvae using biochemical and molecular methods. The results show P. luminescens H06 and X. nematophila All treatments caused significant reductions in the number of O. nipae larvae in a dose-dependent manner. Secondly, the O. nipae immune system recognizes symbiotic bacteria at early and late stages of infection via the induction of C-type lectin. Live symbiotic bacteria significantly inhibit PO activity in O. nipae whereas heat-treated bacteria strongly increase PO activity. Additionally, expression levels of four O. nipae proPhenoloxidase genes following treatment with P. luminescens H06 and X. nematophila All were compared. We found that the expression levels of all proPhenoloxidase genes were significantly down-regulated at all-time points. Similarly, treatments of O. nipae larvae with metabolites benzylideneacetone and oxindole significantly down-regulated the expression of the PPO gene and inhibited PO activity. However, the addition of arachidonic acid to metabolite-treated larvae restored the expression level of the PPO gene and increased PO activity. Our results provide new insight into the roles of symbiotic bacteria in countering the insect phenoloxidase activation system.

Insect pests cause significant agricultural and economic losses to crops worldwide due to their destructive activities. Pesticides are designed to be poisonous and are intentionally released into the environment to combat the menace caused by these noxious pests. To survive, these insects can resist toxic substances introduced by humans in the form of pesticides. According to recent findings, microbes that live in insect as symbionts have recently been found to protect their hosts against toxins. Symbioses that have been formed are between the pests and various microbes, a defensive mechanism against pathogens and pesticides. Insects' guts provide unique conditions for microbial colonization, and resident bacteria can deliver numerous benefits to their hosts. Insects vary significantly in their reliance on gut microbes for basic functions. Insect digestive tracts are very different in shape and chemical properties, which have a big impact on the structure and composition of the microbial community. Insect gut microbiota has been found to contribute to feeding, parasite and pathogen protection, immune response modulation, and pesticide breakdown. The current review will examine the roles of gut microbiota in pesticide detoxification and the mechanisms behind the development of resistance in insects to various pesticides. To better understand the detoxifying microbiota in agriculturally significant pest insects, we provided comprehensive information regarding the role of gut microbiota in the detoxification of pesticides. [ABSTRACT FROM AUTHOR]