Community Structure Of Microbiota Research Articles

Structural and functional differences of gut microbiota in Pomacea canaliculata from different geographical locations and habitats.

Gut microbiota is related to host fitness, and influenced by geographical locations and habitats. Pomacea canaliculata is a malignant invasive alien snail that threatens agricultural production and ecosystem functions worldwide. Clarifying the general rules of the gut microbial community structure and function of the snails in different geographical locations and habitats is of great significance for understanding their invasion at different spatial scales. This study used high-throughput sequencing technology to compare and analyze the differences in community structure and function of gut microbiota in P. canaliculata from five geographical locations (Liuzhou, Yulin, Nanning, Wuzhou, and Hezhou) and three different habitats (pond, paddy field, and ditch) in Guangxi Province. The results showed that intestinal microbial alpha diversity of P. canaliculata was higher in Liuzhou, Yulin, lower in Nanning, Wuzhou, Hezhou, and higher in ponds compared with paddy fields and ditches. The dominant phyla of gut microbiota in snails were Firmicutes, Cyanobacteria, Proteobacteria, Fusobacteriota, Bacteroidota, and the dominant genus was Lactococcus. The community structure of gut microbiota in snails varied significantly across different geographical locations and habitats, and the phyla Firmicutes, Cyanobacteria had significantly higher relative abundance in snails collected from Nanning and Yulin, respectively. Moreover, the relative abundance of gut functional microbiota associated with human disease in P. canaliculata was significantly affected by geographical locations and habitats, and with the highest abundance in ponds. However, the relative abundance of functional microbiota related to metabolism, genetic information processing, organizational system, environmental information processing, and cellular processes were only significantly affected by geographical locations. Collectively, geographical locations and habitats had significantly different effects on the community structure and function of gut microbiota in P. canaliculata, and the greater differences were caused by geographical locations rather than by habitats.

Influence of Biofloc Technology and Continuous Flow Systems on Aquatic Microbiota and Water Quality in Japanese Eel Aquaculture

Biofloc technology (BFT) systems heavily rely on microbiota to mitigate ammonia toxicity and manage essential nutrient cycling. Understanding the diversity and functional role of microbiota within BFT-applied aquaculture systems is crucial for ensuring sustainable operations. Though some studies exist on BFT microbiota, research on microbial differences in Japanese eel aquaculture is still limited, hindering the wider application of BFT systems. In this study, we analyzed the characteristics of water quality factors and microbiota in Japanese eel (Anguilla japonica) breeding water, applying the BFT system. Using a metabarcoding approach, the diversity and community structure of aquatic microbiota were compared between BFT and continuous flow (CF) systems. The pH was significantly higher in CF water, while total ammonia nitrogen (TAN) and nitrite (NO2−-N) was higher in BFT water. Alpha diversity was significantly higher in BFT compared to CF systems, and it was correlated significantly with pH and TAN. In both BFT and CF water, the phyla Proteobacteria and Bacteroidota were found to be the most abundant. In the BFT water, a diverse array of bacterial taxa, including BFT-specific clades, were consistently present, while the microbiota in CF water was more variable and contained fewer specific taxa. In addition, bacterial functions related to nitrate reduction, sulfur compound oxidation, and chitinolysis were significantly more abundant in BFT than in CF systems. These findings highlight differences in water quality and microbiota between aquaculture systems, which can inform future research on the use of BFT for sustainable fish farming.

PSLBI-13 Characterizing the seminal microbiota in mature rams managed on divergent planes of nutrition, and in their male offspring

Abstract Increasing evidence suggests that the bovine semen harbors commensal microbiota, and it may be important in male fertility. However, much is unknown regarding the seminal microbiota, its evolution and factors shaping this community in sheep. The objective of this study was to evaluate whether managing the rams on divergent planes of nutrition can influence the seminal microbiota in mature rams and their male offspring. For this, Rambouillet rams (n = 24; 1.5 to 4 yr old) were randomly assigned to one of three groups: 1) a positive plane of nutrition [target 12% gain in their initial body weight (BW; POS, n = 8); 2) a maintenance [maintain the initial BW (MNT, n = 8)]; and 3) a negative [target a 12% reduction in BW (NEG; n = 8)] plane of nutrition over 84 d. The rams were individually housed and fed a common diet, and feed allocations were adjusted weekly. Following the 84-d feeding period, these rams were used to breed 240 mature Rambouillet ewes (1:10 ram:ewe ratio) over 28 d of breeding. After lambing, the male lambs were maintained on a similar diet for 124 d. Semen samples from the mature rams (F0 generation) were collected on d 28, 56, and 84, and semen samples from offspring rams (F1 generation) were collected at 330 d of age via electroejaculation. Genomic DNA was extracted from semen samples and the seminal microbiota characterized using 16S rRNA gene (V3-V4) sequencing. Several environmental and negative extraction controls were also sequenced to assess potential contaminants in these samples. Overall community structure of the seminal microbiota in F0 rams was not affected by the divergent planes of nutrition (PERMANOVA: P ≥ 0.05) or by sampling time (P > 0.05). Microbial richness (observed ASVs) was also not different (P > 0.05) between the three groups of F0 rams at any of the sampling time points; however, microbial diversity (P < 0.05) in the F0 rams was influenced by sire dietary treatment. Microbial community structure, richness and diversity did not differ among the POS, MNT and NEG F1 generation rams (P > 0.05). Actinobacteriota, Firmicutes, Bacteroidota, Proteobacteria and Fusobacteriota were the most abundant phyla. Only Bacteroidota abundance was significantly (P < 0.05) different between three F0 groups, with POS rams had greater (18.6%) of this phylum on d 28 as compared with MNT (2.1%) and NEG (5.1%). The predominant genera identified included Fastidiosipila, Corynebacterium, Trueperella, Arthrobacter, Dietzia, Bifidobacterium, Streptobacillus, Ornithinimicrobium, and Porphyromonas, and their relative abundance was not affected (P > 0.05) by the dietary treatment in F0 rams. Overall, rams managed on positive and negative planes of nutrition harbored similar seminal microbiota to control rams, and divergent nutrition also had no effect on the offspring ram seminal microbiota.

PSV-12 Evaluating the effect of lyoprotectants in preserving community structure and bacterial viability in the lyophilized fecal community of pigs

Abstract Fecal microbiota transplantation (FMT) stands as a potential method for enhancing pig health. FMT has progressed from the basic delivery of fresh stool, to lyophilized preparations which is beneficial for sample storage, but can damage bacterial cells. This has necessitated the need for lyoprotectants to preserve bacterial viability. However, the ideal lyoprotectant for retaining community structure and viability remains unclear. Furthermore, DNA-based microbial profiling methods such as 16S rRNA and metagenomics sequencing do not distinguish live and dead bacteria, prompting the need for other methods of distinguishing live and dead members of the bacterial community after lyophilization. PMA-seq (propidium monoazide treatment followed by 16S rRNA gene amplicon sequencing) has been shown to differentiate between live and dead bacteria in microbial communities. To refine FMT in pigs, we sought to ascertain the impact of four lyoprotectants on bacterial viability and fecal microbiome structure. Fecal samples from six pigs were treated with mannitol, maltodextrin, trehalose, maltodextrin-trehalose mix (2.5% each) as lyoprotectants or left untreated. Lyophilized fecal samples underwent PMA treatment prior to 16S rRNA sequencing to capture the active community, and total and viable bacterial cells number were quantified. One-way ANOVA was used to analyze total and viable bacterial cells, and richness, while PERMANOVA was used to compare beta diversity. Results revealed no initial differences in total, viable bacteria, or community structure (P > 0.05). However, untreated, and mannitol-treated groups showed significantly reduced total and viable bacterial cells post-lyophilization (P < 0.01), unlike the other lyoprotectants (P > 0.05). Mannitol and the untreated group also displayed decreased richness and beta diversity shifts (P < 0.05). Overall, lyophilization with mannitol or no lyoprotectant notably impacted pig fecal microbiota community structure, while maltodextrin, trehalose, and maltodextrin-trehalose effectively preserved bacterial viability and structure.

Leaf Shelters Facilitate the Colonisation of Arthropods and Enhance Microbial Diversity on Plants.

Shelter-building insects are important ecosystem engineers, playing critical roles in structuring arthropod communities. Nonetheless, the influence of leaf shelters and arthropods on plant-associated microbiota remains largely unexplored. Arthropods that visit or inhabit plants can contribute to the leaf microbial community, resulting in significant changes in plant-microbe interactions. By artificially constructing leaf shelters, we provide evidence that shelter-building insects influence not only the arthropod community structure but also impact the phyllosphere microbiota. Leaf shelters exhibited higher abundance and richness of arthropods, changing the associated arthropod community composition. These shelters also altered the composition and community structure of phyllosphere microbiota, promoting greater richness and diversity of bacteria at the phyllosphere. In leaf shelters, microbial diversity positively correlated with the richness and diversity of herbivores. These findings demonstrate the critical role of leaf shelters in structuring both arthropod and microbial communities through altered microhabitats and species interactions.

Effects of dietary restriction and one-carbon metabolite supplementation during the first 63 days of gestation on the maternal gut, vaginal, and blood microbiota in cattle

BackgroundMaternal diet quality and quantity have significant impacts on both maternal and fetal health and development. The composition and function of the maternal gut microbiome is also significantly influenced by diet; however, little is known about the impact of gestational nutrient restriction on the bovine maternal microbiome during early gestation, which is a critical stage for maternal microbiome-mediated fetal programming to take place. The objective of the present study was to evaluate the impacts of diet restriction and one-carbon metabolite (OCM) supplementation during early gestation on maternal ruminal, vaginal, and blood microbiota in cattle. Thirty-three beef heifers (approx. 14 months old) were used in a 2 × 2 factorial experiment with main factors of target gain (control [CON]; targeted 0.45 kg/d gain vs restricted [RES]; targeted − 0.23 kg/d gain), and OCM supplementation (+ OCM vs − OCM; n = 8/treatment; except n = 9 for RES–OCM). Heifers were individually fed, starting treatment at breeding (d 0) and concluding at d 63 of gestation. Ruminal fluid and vaginal swabs were collected on d − 2, d 35, and d 63 (at necropsy) and whole blood was collected on d 63 (necropsy). Bacterial microbiota was assessed using 16S rRNA gene (V3–V4) sequencing.ResultsOverall ruminal microbiota structure was affected by gain, OCM, time, and their interactions. The RES heifers had greater microbial richness (observed ASVs) but neither Shannon nor Inverse Simpson diversity was significantly influenced by gain or OCM supplementation; however, on d 63, 34 bacterial genera showed differential abundance in the ruminal fluid, with 25 genera enriched in RES heifers as compared to CON heifers. In addition, the overall interaction network structure of the ruminal microbiota changed due to diet restriction. The vaginal microbiota community structure was influenced by gain and time. Overall microbial richness and diversity of the vaginal microbiota steadily increased as pregnancy progressed. The vaginal ecological network structure was distinctive between RES and CON heifers with genera-genera interactions being intensified in RES heifers. A relatively diverse bacterial community was detected in blood samples, and the composition of the blood microbiota differed from that of ruminal and vaginal microbiota.ConclusionRestricted dietary intake during early gestation induced significant alterations in the ruminal microbiota which also extended to the vaginal microbiota. The composition of these two microbial communities was largely unaffected by OCM supplementation. Blood associated microbiota was largely distinctive from the ruminal and vaginal microbiota.

Microbiome and Metabolome Restoration After Administration of Fecal Microbiota, Live-jslm (REBYOTA®) for Preventing Recurrent Clostridioides difficile Infection.

Microbiota-based treatments are effective in preventing recurrent Clostridioides difficile infection (rCDI). Fecal microbiota, live-jslm (REBYOTA®; RBL, previously RBX2660) was shown to prevent rCDI in a phase 3, randomized, double-blinded placebo controlled clinical trial (PUNCH™ CD3). Stool samples from participants in PUNCH™ CD3 who received a single blinded dose of rectally administered RBL or placebo were sequenced to determine microbial community composition and calculate the Microbiome Health Index for post-antibiotic dysbiosis (MHI-A). The composition of bile acids (BAs) in the same samples was quantified by liquid chromatography mass spectrometry. Relationships between BA composition and microbiota community structure and correlations with treatment outcomes were assessed. Before administration, Gammaproteobacteria and Bacilli dominated the microbiota community and primary BAs were more prevalent than secondary BAs. Clinical success after administration correlated with shifts to predominantly Bacteroidia and Clostridia, a significant increase in MHI-A, and a shift from primary to secondary BAs. Several microbiota and BA changes were more extensive in RBL-treated responders compared to placebo-treated responders, and microbiota changes correlated with BA changes. Clinical response and RBL administration were associated with significant restoration of microbiota and BA composition. NCT03244644.

Changes in bumblebee queen gut microbiotas during and after overwintering diapause.

Bumblebees are key pollinators with gut microbiotas that support host health. After bumblebee queens undergo winter diapause, which occurs before spring colony establishment, their gut microbiotas are disturbed, but little is known about community dynamics during diapause itself. Queen gut microbiotas also help seed worker microbiotas, so it is important that they recover post-diapause to a typical community structure, a process that may be impeded by pesticide exposure. We examined how bumblebee queen gut microbiota community structure and metabolic potential shift during and after winter diapause, and whether post-diapause recovery is affected by pesticide exposure. To do so, we placed commercial Bombus impatiens queens into diapause, euthanizing them at 0, 2 and 4 months of diapause. Additionally, we allowed some queens to recover from diapause for 1 week before euthanasia, exposing half to the common herbicide glyphosate. Using whole-community, shotgun metagenomic sequencing, we found that core bee gut phylotypes dominated queen gut microbiotas before, during and after diapause, but that two phylotypes, Schmidhempelia and Snodgrassella, ceased to be detected during late diapause and recovery. Despite fluctuations in taxonomic community structure, metabolic potential remained constant through diapause and recovery. Also, glyphosate exposure did not affect post-diapause microbiota recovery. However, metagenomic assembly quality and our ability to detect microbial taxa and metabolic pathways declined alongside microbial abundance, which was substantially reduced during diapause. Our study offers new insights into how bumblebee queen gut microbiotas change taxonomically and functionally during a key life stage and provides guidance for future microbiota studies in diapausing bumblebees.

Landscape of the Lumbar Cartilaginous End Plate Microbiota and Metabolites in Patients with Modic Changes.

Modic changes (MCs), vertebral end plate and bone marrow damage observed by magnetic resonance imaging, are an independent risk factor for low back pain. The compositions of and interaction between microbiota and metabolites in the lumbar cartilaginous end plates (LCEPs) of patients with MCs have not been identified. Patients with lumbar disc degeneration who were undergoing lumbar spinal fusion surgery were recruited between April 2020 and April 2021. LCEPs were collected for 16S rRNA sequencing and liquid chromatography-mass spectrometry (LC/MS)-based targeted metabolomic profiling. Of the 54 patients recruited, 24 had no MCs and 30 had changes classified as Modic type 2 or 3. The primary goal was to identify specific genera of microbiota associated with MCs, and secondary goals included investigating differences in metabolites between patients with and without MCs and exploring the correlation between these metabolites and microorganisms. Investigation of the microbiota community structure revealed that both alpha diversity and beta diversity were significantly different between patients with and without MCs, and the abundances of 26 genera were significantly different between these 2 groups. Metabolomic analysis revealed that 26 metabolites were significantly different between the 2 groups. The unsaturated fatty acid pathway was found to be the main pathway related to MCs. Multiomic correlation analysis suggested that Caulobacteraceae (unclassified) and Mycobacterium, Clostridium, Blautia, and Bifidobacterium at the genus level were linked to dysregulation of fatty acid metabolism, contributing to the pathogenesis of MCs. Our study represents a foundational effort to examine the landscape of the microbiota and metabolites in patients with MCs, informing future studies on the pathogenesis of and targeted therapy for MCs. Prognostic Level II. See Instructions for Authors for a complete description of levels of evidence.

Mosquito Gut Microbiota: A Review.

Mosquitoes are vectors of many important human diseases. The prolonged and widespread use of insecticides has led to the development of mosquito resistance to these insecticides. The gut microbiota is considered the master of host development and physiology; it influences mosquito biology, disease pathogen transmission, and resistance to insecticides. Understanding the role and mechanisms of mosquito gut microbiota in mosquito insecticide resistance is useful for developing new strategies for tackling mosquito insecticide resistance. We searched online databases, including PubMed, MEDLINE, SciELO, Web of Science, and the Chinese Science Citation Database. We searched all terms, including microbiota and mosquitoes, or any specific genera or species of mosquitoes. We reviewed the relationships between microbiota and mosquito growth, development, survival, reproduction, and disease pathogen transmission, as well as the interactions between microbiota and mosquito insecticide resistance. Overall, 429 studies were included in this review after filtering 8139 search results. Mosquito gut microbiota show a complex community structure with rich species diversity, dynamic changes in the species composition over time (season) and across space (environmental setting), and variation among mosquito species and mosquito developmental stages (larval vs. adult). The community composition of the microbiota plays profound roles in mosquito development, survival, and reproduction. There was a reciprocal interaction between the mosquito midgut microbiota and virus infection in mosquitoes. Wolbachia, Asaia, and Serratia are the three most studied bacteria that influence disease pathogen transmission. The insecticide resistance or exposure led to the enrichment or reduction in certain microorganisms in the resistant mosquitoes while enhancing the abundance of other microorganisms in insect-susceptible mosquitoes, and they involved many different species/genera/families of microorganisms. Conversely, microbiota can promote insecticide resistance in their hosts by isolating and degrading insecticidal compounds or altering the expression of host genes and metabolic detoxification enzymes. Currently, knowledge is scarce about the community structure of mosquito gut microbiota and its functionality in relation to mosquito pathogen transmission and insecticide resistance. The new multi-omics techniques should be adopted to find the links among environment, mosquito, and host and bring mosquito microbiota studies to the next level.

Energy reserves and gut microbiota of marine mussels under combined exposure to pathogens and predation risk

In recent years, environmental issues such as eutrophication have led to proliferation of pathogenic bacteria and deterioration of aquaculture environmental water quality. However, the impact of pathogenic bacteria on the mussel-predator food chain in the marine environment is not yet fully understood. In this experiment, mussels were cultured with crabs (Charybdis japonica) for 14 days. Then crabs were retained or removed and pathogenic bacteria (Vibrio alginolyticus) was added and mussels were cultured for 3 days. C. japonica and V. alginolyticus were used as two stressors to explore their effects on energy content and gut microbiota of Mytilus coruscus. The metabolic parameters (protein, carbohydrate and lipid contents) of the gills and digestive glands, and the gut microbiota of the mussels were measured. The study found that presence of predation risk significantly increased the content of protein but decreased carbohydrate and lipid contents in the gills at 14th day. After 17 days of exposure, the presence of predation risk significantly reduced protein content but significantly increased lipid content in gills of mussel. The protein content in digestive gland was significantly increased, but the lipid content was significantly decreased. The presence of predation risk and pathogens had interactive effect on the protein content in the digestive glands of mussels. The presence of V. alginolyticus and predation risk affected the community structure, species richness, and diversity of gut microbiota. The most abundant phyla noted were Bacteroidota, Proteobacteria, and Firmicutes, and the most abundant genus noted were Prevotella, Klebsiella and Dialister. Alpha diversity analysis and beta diversity analysis revealed that the presence of V. alginolyticus and predation risk altered the microbial community structure. V. alginolyticus plays a positive role in the harmful microbiota of gut microbiota. The proportion of Bacteroidota and Proteobacteria in intestinal bacteria increased significantly. Linear discriminant analysis effect size (LEfSe) showed that the biomarkers in each group are significantly different at the genus level. Significant correlations between metabolic parameters (protein content and carbohydrate content) and the level of KEGG function prediction (amino acid metabolism and carbohydrate metabolism) were observed in gills and digestive glands. This study highlights the priority of energy responses in mussels under dual predator-pathogen pressure, and also highlights the complexity of predator-pathogen interactions. A better understanding of their impact on the gut microbiota and health of marine mussels will help support the sustainability of mussel aquaculture production.

Microplastics predominantly affect gut microbiota by altering community structure rather than richness and diversity: A meta-analysis of aquatic animals

The impacts of microplastics on the gut microbiota, a crucial component of the health of aquatic animals, remain inadequately understood. This phylogenetically controlled meta-analysis aims to identify general patterns of microplastic effects on the alpha diversity (richness and Shannon index), beta diversity, and community structure of gut microbiota in aquatic animals. Data from 63 peer-reviewed articles on the Web of Science were synthesized, encompassing 424 observations across 31 aquatic species. The analysis showed that microplastics significantly altered the community structure of gut microbiota, with between-group distances being 87.75% higher than within-group distances. This effect was significant even at environmentally relevant concentrations (≤1 mg L−1). However, their effects on richness, Shannon index, and beta diversity (community variation) were found to be insignificant. The study also indicated that the effects of microplastics were primarily dependent on their concentration and size, while the phylogeny of tested species explained limited heterogeneity. Furthermore, variations in gut microbiota alpha diversity, beta diversity, and community structure were correlated with changes in antioxidant enzyme activities from the liver and hepatopancreas. This implies that gut microbiota attributes of aquatic animals may provide insights into host antioxidant levels. In summary, this study illuminates the impacts of microplastics on the gut microbiota of aquatic animals and examines the implications of these effects for host health. It emphasizes that microplastics mainly alter the community structure of gut microbiota rather than significantly affecting richness and diversity.

Impact of Nutrient Enrichment on Community Structure and Co-Occurrence Networks of Coral Symbiotic Microbiota in Duncanopsammia peltata: Zooxanthellae, Bacteria, and Archaea.

Symbiotic microorganisms in reef-building corals, including algae, bacteria, archaea, fungi, and viruses, play critical roles in the adaptation of coral hosts to adverse environmental conditions. However, their adaptation and functional relationships in nutrient-rich environments have yet to be fully explored. This study investigated Duncanopsammia peltata and the surrounding seawater and sediments from protected and non-protected areas in the summer and winter in Dongshan Bay. High-throughput sequencing was used to characterize community changes, co-occurrence patterns, and factors influencing symbiotic coral microorganisms (zooxanthellae, bacteria, and archaea) in different environments. The results showed that nutrient enrichment in the protected and non-protected areas was the greatest in December, followed by the non-protected area in August. In contrast, the August protected area had the lowest nutrient enrichment. Significant differences were found in the composition of the bacterial and archaeal communities in seawater and sediments from different regions. Among the coral symbiotic microorganisms, the main dominant species of zooxanthellae is the C1 subspecies (42.22-56.35%). The dominant phyla of bacteria were Proteobacteria, Cyanobacteria, Firmicutes, and Bacteroidota. Only in the August protected area did a large number (41.98%) of SAR324_cladeMarine_group_B exist. The August protected and non-protected areas and December protected and non-protected areas contained beneficial bacteria as biomarkers. They were Nisaea, Spiroplasma, Endozoicomonas, and Bacillus. No pathogenic bacteria appeared in the protected area in August. The dominant phylum in Archaea was Crenarchaeota. These symbiotic coral microorganisms' relative abundances and compositions vary with environmental changes. The enrichment of dissolved inorganic nitrogen in environmental media is a key factor affecting the composition of coral microbial communities. Co-occurrence analysis showed that nutrient enrichment under anthropogenic disturbances enhanced the interactions between coral symbiotic microorganisms. These findings improve our understanding of the adaptations of coral holobionts to various nutritional environments.

Dairy effluent applications to a pasture enhance soil fertility and microbial activity without impacting soil bacterial and fungal community composition

Farm dairy effluents (FDE) from washing the milking parlor contain manure, urine, and chemicals and constitute a large amount of wastewater. Applying FDE as soil fertilizers to pastures can enhance forage yield and improve soil nutrient status. Since the dairy industry is increasingly attempting to maximize returns through better utilization of forage with lesser inputs, there is demand for a supply of FDE as fertilizers. Nevertheless, the impact of this practice on soil microbiota remains largely unexplored. It must be studied before large-scale soil disposal to avoid diminishing microbial diversity or enhancing pathogen abundance. This study evaluated the effects of applying lagoon-stored (Lagoon) and raw dairy effluents (Raw) at a rate of 50 kg N ha−1 in four equal doses, in comparison to urea fertilization, on soil fertility and the activity, abundance, and community structure of soil microbiota. Raw was obtained after solid separation, and Lagoon corresponds to the Raw stationed in a two-lagoon system. Microbial activity was assessed as basal respiration, potentially mineralizable N, potential nitrification activity, and enzymatic activities. The catabolic activity of the microbial community was evaluated using Biolog Ecoplates™. Bacterial and fungal community composition and diversity were analyzed through amplicon sequencing of 16S rRNA and ITS2. The application of FDE benefited soil fertility and microbial activity. Lagoon had the most potent effects on soil available P and extractable K+, Na+, Mg2+ and Ca2+. Soil treated with Raw displayed higher microbial activities, such as dehydrogenase, basal respiration, urease, and potentially mineralizable N, than the other treatments. FDE did not significantly alter the microbial composition, abundance, or functional diversity. In conclusion, in this short-term trial, despite changes in soil chemical properties and microbial activity, the composition and diversity of the bacterial and fungal communities remained unaffected by FDE irrigation.

Integrating functional metagenomics to decipher microbiome-immune interactions.

Microbial metabolites can be viewed as the cytokines of the microbiome, transmitting information about the microbial and metabolic environment of the gut to orchestrate and modulate local and systemic immune responses. Still, many immunology studies focus solely on the taxonomy and community structure of the gut microbiota rather than its functions. Early sequencing-based microbiota profiling approaches relied on PCR amplification of small regions of bacterial and fungal genomes to facilitate identification of the microbes present. However, recent microbiome analysis methods, particularly shotgun metagenomic sequencing, now enable culture-independent profiling of microbiome functions and metabolites in addition to taxonomic characterization. In this review, we showcase recent advances in functional metagenomics methods and applications and discuss the current limitations and potential avenues for future development. Importantly, we highlight a few examples of key areas of opportunity in immunology research where integrating functional metagenomic analyses of the microbiome can substantially enhance a mechanistic understanding of microbiome-immune interactions and their contributions to health and disease states.

Genetic hypogonadal (Gnrh1hpg) mouse model uncovers influence of reproductive axis on maturation of the gut microbiome during puberty.

The gut microbiome plays a key role in human health and gut dysbiosis is linked to many sex-specific diseases including autoimmune, metabolic, and neurological disorders. Activation of the hypothalamic-pituitary-gonadal (HPG) axis during puberty leads to sexual maturation and development of sex differences through the action of gonadal sex steroids. While the gut microbiome also undergoes sex differentiation, the mechanisms involved remain poorly understood. Using a genetic hypogonadal (hpg) mouse model, we sampled the fecal microbiome of male and female wild-type and hpg mutant mice before and after puberty to determine how microbial taxonomy and function are influenced by age, sex, and the HPG axis. We showed that HPG axis activation during puberty is required for sexual maturation of the gut microbiota composition, community structure, and metabolic functions. We also demonstrated that some sex differences in taxonomic composition and amine metabolism developed independently of the HPG axis, indicating that sex chromosomes are sufficient for certain sex differences in the gut microbiome. In addition, we showed that age, independent of HPG axis activation, led to some aspects of pubertal maturation of the gut microbiota community composition and putative functions. These results have implications for microbiome-based treatments, indicating that sex, hormonal status, and age should be considered when designing microbiome-based therapeutics.

Dissecting the ecological risks of sulfadiazine degradation intermediates under different advanced oxidation systems: From toxicity to the fate of antibiotic resistance genes

The incomplete degradation of antibiotics in water can produce intermediates that carry environmental risks and thus warrant concerns. In this study, the degradation of high concentrations of antibiotic sulfadiazine (SDZ) by advanced oxidation processes that leverage different reactive oxide species was systematically evaluated in terms of the influence of different degradation intermediates on the propagation of antibiotic resistance genes (ARGs). The ozone, persulfate, and photocatalytic oxidation systems for SDZ degradation are dominated by ozone, direct electron transfer, and singlet oxygen, hole, and superoxide radicals, respectively. These processes produce 15 intermediates via six degradation pathways. Notably, it was determined that three specific intermediates produced by the ozone and persulfate systems were more toxic than SDZ. In contrast, the photocatalytic system did not produce any intermediates with toxicity exceeding that of SDZ. Microcosm experiments combined with metagenomics confirmed significant changes in microbiota community structure after treatment with SDZ and its intermediates, including significant changes in the abundance of Flavobacterium, Dungenella, Archangium, and Comamonas. This treatment also led to the emergence of sulfonamide ARGs. The total abundance of sulfonamide ARGs was found to be positively correlated with residual SDZ concentration, with the lowest total abundance observed in the photocatalytic system. Additionally, the correlation analysis unveiled microbiota carrying sulfonamide ARGs.

Analysis of Gut Microbiota as a Diagnostic Biomarker for Lung Adenocarcinoma with Qi-Deficiency and Phlegm-Turbid Stagnation

Background: In lung adenocarcinoma (LUAD), Qi-deficiency and Phlegm-turbid stagnation (QP) are the most prevalent Traditional Chinese medicine (TCM) syndrome. Methods: Herein, we collected 90 fecal samples (Healthy individual (H): 30; other syndrome (O): 30; QP: 30) and explored the composition and diversity of gut microbiota in LUAD patients with QP syndrome using 16s-rRNA sequencing. Then, we identified biomarkers for QP syndrome in LUAD patients with linear discriminant analysis (LDA) effect size (LEfSe) and applied logistic regression analysis to construct a diagnostic model evaluated with the area under the receiver operating characteristic curve (AUC) and validated with data from metagenomics. Results: The α diversity and β diversity revealed that the microbiota community structure in LUAD patients with QP syndrome was different from that with healthy individuals and LUAD patients with other syndromes. At the phylum level, the QP group had more abundance of Bacteroidetes and less Proteobacteria than the O group. At the genus level, the abundance of 4 genera (Bacteroides, Parabacteroides, Prevotella, and Flavonifractor) was different between the QP group and O group. Moreover, LEfSe indicated that those 4 genera might be the biomarkers for LUAD patients with QP syndrome. Then, we used those 4 genera to develop a diagnostic model. The AUC based on 16s-rRNA sequencing and metagenomics was 0.989 and 1, respectively. Conclusion: A diagnostic model was developed, which would be an available tool for the clinical diagnosis of LUAD with QP syndrome.

Exploratory analysis of gut microbiota differences in patients with bronchial asthma of different inflammatory types

Objective: To observe the characteristics and differences of gut microbiota in asthma patients with different inflammatory types through metagenomic analysis. Methods: Adults aged ≥18 years who visited the Respiratory Clinic of Peking University Third Hospital from August 1, 2021 to August 31, 2022 and were primarily diagnosed with asthma were selected as the study subjects. Finally, 29 patients with stable asthma were included. Fresh fecal samples were collected and the fecal DNA was extracted for high-throughput 16sRNA sequencing of gut microbiota. The diversity and community structure of gut microbiota in different groups of asthma patients were compared, and the species differences were analyzed through random forest and LEfSe analysis. Results: There were sex-based differences in asthma patients with different types of inflammation, and the proportion of female patients was higher in neutrophilic asthma patients (χ2=4.14, P=0.042). There was no significant intergroup difference in the alpha diversity of gut microbiota among asthma patients with different inflammatory types, but there were significant differences in the microbiome. Patients with neutrophilic asthma had higher relative abundance of Bacillales (P=0.029) and Oscillospiraceae (P=0.015). In species LEfSe analysis, patients with eosinophilic asthma had a higher relative abundance of fungi. Conclusion: There are intergroup differences in the gut microbiota of asthma patients with different inflammation types, and fungi are biomarkers that distinguish the differences in gut microbiota between patients with eosinophilic asthma and neutrophilic asthma.

Type-2 diabetes mellitus enhances Klebsiella pneumoniae pathogenesis.

Klebsiella pneumoniae is an opportunistic pathogen and an important cause of pneumonia, bacteremia, and urinary tract infection. K. pneumoniae infections are historically associated with diabetes mellitus. There is a fundamental gap in our understanding of how diabetes mellitus, specifically type 2 diabetes, influences K. pneumoniae pathogenesis. K. pneumoniae pathogenesis is a multifactorial process that often begins with gut colonization, followed by an escape from the gut to peripheral sites, leading to host damage and infection. We hypothesized that type 2 diabetes enhances K. pneumoniae pathogenesis. To test this, we used well-established mouse models of K. pneumoniae colonization and lung infection in conjunction with a mouse model of spontaneous type 2 diabetes mellitus (T2DM). We show that T2DM enhances susceptibility to both K. pneumoniae colonization and infection. The enhancement of gut colonization is dependent on T2DM-induced modulation of the gut microbiota community structure. In contrast, lung infection is exacerbated by the increased availability of amino acids in the lung, which is associated with higher levels of vascular endothelial growth factor. These data lay the foundation for mechanistic interrogation of the relationship between K. pneumoniae pathogenesis and type 2 diabetes mellitus, and explicitly establish T2DM as a risk factor for K. pneumoniae disease.