Microbiome Community Research Articles

Diversity of gut microbiome in Rocky Mountainsnail across its native range.

The animal gut microbiome is often a key requirement for host nutrition, digestion, and immunity, and can shift in relation to host geography and environmental factors. However, ecological drivers of microbiome community assembly across large geographic ranges have rarely been examined in invertebrates. Oreohelix strigosa (Rocky Mountainsnail) is a widespread land snail found in heterogeneous environments across the mountainous western United States. It is ideally suited for biogeography studies due to its broad distribution, low migration, and low likelihood of passive transport via other animals. This study aims to uncover large-scale geographic shifts in the composition of O. strigosa gut microbiomes by using 16S rRNA gene sequencing on samples from across its native range. Additionally, we elucidate smaller-scale microbiome variation using samples collected only within Colorado. Results show that gut microbiomes vary significantly across broad geographic ranges. Several possible ecological drivers, including soil and vegetation composition, habitat complexity, habitat type, and human impact, collectively explained 27% of the variation across Coloradan O. strigosa gut microbiomes. Snail gut microbiomes show more similarity to vegetation than soil microbiomes. Gut microbial richness was highest in the rocky habitats and increased significantly in the most disturbed habitats (low complexity, high human impact), potentially indicating signs of dysbiosis in the snails' gut microbiomes. These small-scale environmental factors may be driving changes in O. strigosa gut microbiome composition seen across large-scale geography. This knowledge will also help us better understand how microbial associations influence species survival in diverse environments and aid wildlife conservation efforts.

Total Astragalus saponins can reverse type 2 diabetes mellitus-related intestinal dysbiosis and hepatic insulin resistance in vivo.

Intestinal flora homeostasis in rats with type 2 diabetes mellitus (T2DM) was evaluated to explore the effects of total Astragalus saponins (TAS) on hepatic insulin resistance (IR). Six-week-old male Sprague-Dawley rats were fed high-fat and high-sugar diet for 4 weeks and intraperitoneally injected with streptozotocin to induce T2DM, and they were then randomly divided into control, model, metformin, and TAS groups. Stool, serum, colon, and liver samples were collected after 8 weeks of drug administration for relevant analyses. TAS reduced fasting blood glucose, 2-hour postprandial blood glucose, area under the curve of oral glucose tolerance test, glycated serum protein, homeostasis model assessment of insulin resistance, total cholesterol, triglyceride, and low-density lipoprotein cholesterol levels in T2DM rats but increased insulin, C-peptide, and high-density lipoprotein cholesterol levels. Moreover, TAS improved the morphology and structure of liver and colon tissues and improved the composition of the intestinal microbiome and bacterial community structure at different taxonomic levels. In addition, TAS increased the protein expression of hepatic IRS-1, PI3K, PDK1, and p-AKT and decreased the protein expression of p-GSK-3β. Meanwhile, TAS increased the mRNA expression of liver PDK1, PI3K, and GS and decreased the mRNA expression of GSK-3β. TAS can ameliorate T2DM-related abnormal glucose and blood lipid metabolism, intestinal dysbiosis, and IR.

The effects of enrofloxacin exposure on responses to oxidative stress, intestinal structure and intestinal microbiome community of largemouth bass (Micropterus salmoides)

Antibiotic residues in the aquaculture environments may lead to antibiotic resistance, and potentially exert adverse effects on health of the non-target organisms and humans. In order to evaluate the effect of enrofloxacin of environmental concentrations on largemouth bass (Micropterus salmoides). Two hundred and seventy largemouth basses (with an average weight of 7.88 ± 0.60 g) were randomly divided into three groups, and separately exposed to 0, 1, 100 μg/L enrofloxacin (Control, ENR1, ENR100) for 30 days to detect the effect of enrofloxacin on the growth performance, oxidative stress, intestinal microbiota structure, inflammatory response and structure of the intestine. The results showed that ENR significantly reduced the final body weight (FBW) and weight gain rate (WGR), and increased feed conversion ratio (FCR) (P < 0.05). The histopathological analysis revealed that the villus width and muscular thickness of anterior intestine were significantly decreased with the increasing of enrofloxacin concentration. The activity of SOD was significantly increased at enrofloxacin stress, while CAT and POD activity were significantly decreased compared to control group (P < 0.05). The activities of lysozyme (LZM), alkaline phosphatase (AKP) and peroxidase (POD) in ENR1 was higher than that of control and ENR100 groups. Enrofloxacin treatment up-regulated the expression IL-1β and TNF-α, and down-regulated IL-10, and decreasing the expression level ZO-1, claudin-1, and occludin. Furthermore, the enrofloxacin treatment significantly decreased the intestinal bacterial diversity (P < 0.05). Exposure to 100 μg/L enrofloxacin obviously increased the relative abundance of Bacteroidota, Myxococcota, and Zixibacteria of fish gut, and reduced Firmicutes; 1 μg/L enrofloxacin considerably increased Bacteroidota, Myxococcota, and Actinobacteria, and reduced Firmicutes. The relative abundance of DTB120 and Elusimicrobiota was positively correlated with the occludin and claudin-1 gene. Taken together, exposure to enrofloxacin inhibited the growth of largemouth bass, influenced intestinal health, and induced dysbiosis of the intestinal microbiota.

Aedes aegypti microbiome composition covaries with the density of Wolbachia infection

BackgroundWolbachia is a widespread bacterial endosymbiont that can inhibit vector competency when stably transinfected into the mosquito, Aedes aegypti, a primary vector of the dengue virus (DENV) and other arboviruses. Although a complete mechanistic understanding of pathogen blocking is lacking, it is likely to involve host immunity induction and resource competition between Wolbachia and DENV, both of which may be impacted by microbiome composition. The potential impact of Wolbachia transinfection on host fitness is also of importance given the widespread release of mosquitos infected with the Drosophila melanogaster strain of Wolbachia (wMel) in wild populations. Here, population-level genomic data from Ae. aegypti was surveyed to establish the relationship between the density of wMel infection and the composition of the host microbiome.ResultsAnalysis of genomic data from 172 Ae. aegypti females across six populations resulted in an expanded and quantitatively refined, species-level characterization of the bacterial, archaeal, and fungal microbiome. This included 844 species of bacteria across 23 phyla, of which 54 species were found to be ubiquitous microbiome members across these populations. The density of wMel infection was highly variable between individuals and negatively correlated with microbiome diversity. Network analyses revealed wMel as a hub comprised solely of negative interactions with other bacterial species. This contrasted with the large and highly interconnected network of other microbiome species that may represent members of the midgut microbiome community in this population.ConclusionOur bioinformatic survey provided a species-level characterization of Ae. aegypti microbiome composition and variation. wMel load varied substantially across populations and individuals and, importantly, wMel was a major hub of a negative interactions across the microbiome. These interactions may be an inherent consequence of heightened pathogen blocking in densely infected individuals or, alternatively, may result from antagonistic Wolbachia-incompatible bacteria that could impede the efficacy of wMel as a biological control agent in future applications. The relationship between wMel infection variation and the microbiome warrants further investigation in the context of developing wMel as a multivalent control agent against other arboviruses.AACNEMfgWJ36enPNiFi-k-Video

Getting to the Root of Tree Soil Microbiome Sampling

Microbiomes play critical roles in host functioning and, therefore, there is increasing interest in the microbiome assembly of plants. However, sampling strategies for long-lived perennial trees need to be standardized to produce robust data that accurately represent the microbiome over time. This issue is currently unresolved because there is little evidence indicating which portion of perennial tree species (e.g., root region or surrounding soil) is the best to sample to produce the most accurate measure of microbiome communities. Our aim was to sample different compartments of a plant's belowground microbiome to identify the optimal sampling strategy to account for the microbial community present. We found that the structure of the microbial community depends most strongly on the environment (site) and compartment of sample collected (bulk soil, rhizosphere, or rhizoplane), rather than the depth or cardinal direction of the sample. We also found that the microbial community increased in diversity with increased distance from the tree within the rhizoplane and rhizosphere. The data presented here provide systematic evidence for a pragmatic and robust sampling regime that was tested and validated across different environments and soil types while controlling for host genotype. This sampling regime will enable effective partitioning of root compartments when studying the microbiome associated with perennial tree species, allowing targeted questions about the microbiome to be explored with greater accuracy.

The Oral Microbiome Profile of Water Polo Players Aged 16-20.

Chlorine has a strong antibacterial property and is the disinfectant most frequently used in swimming pools. Therefore, the microbiota community in the oral cavity of those who practice water sports is assumed to be special due to their regular immersion in water. Adverse changes in the composition of oral cavity microbiota may have serious health consequences. We aimed to compare the oral microbiome between water polo players and non-athletes. We hypothesized that the oral cavity microbiota community differed between water polo players and non-athletes. Altogether, 124 water polo players (62 males and 62 females, aged between 9 and 20 years) and 16 non-athlete youths (control group, eight males and eight females, aged between 16 and 20 years, mean age + SD = 17.1 + 1.4 years) who participated in body structure examinations voluntarily agreed to participate in the study. In a randomly selected subsample of water polo players (n: 29, aged between 16 and 20 years, mean age + SD = 17.3 + 1.0 years), saliva samples were also collected. Saliva samples were collected from all non-athlete youths (n: 16, aged between 16 and 20 years). The oral microbiome was determined from a saliva sample, and DNA was isolated using the QIAmp DNA Blood Mini Kit. The 16S rRNA gene amplicon sequencing method was used to analyze the microbiome community. PCR primers were trimmed from the sequence reads with Cutadapt. R library DADA2 was used to process reads in the abundance analysis. In general, Streptococcus, Veilonella, and Prevotella genera constituted more than 50% of the oral microbiome community in the two participant groups combined (n = 45). The oral microbial profile had significant sexual dimorphism and differed between water polo players and the non-athletes. Compared to females, males had a higher (p < 0.05) relative abundance of the Atopobium (medium effect size) and Pravotella_7 (very large effect size) genera and a lower (p < 0.05) relative abundance of the Fusobacterium (large effect size), Gemella (large effect size), and Streptococcus (large effect size) genera. Compared to non-athletes, water polo players had higher (p < 0.05, medium effect size) relative abundance of the genus Veillonella and lower (p < 0.05, large effect size) relative abundance of the genus Gemella. The results suggest that regular water training can unfavorably alter the composition of the oral microbial community.

PSXI-11 Microbiome Taxa Involved in Carbohydrate Breakdown and Synthesis of Short Chain Fatty Acids in the Piglet Gut

Abstract The gut microbiome plays a critical role in swine health and growth, and diet is a key factor which shapes microbiome communities. In particular, the fermentation of indigestible carbohydrates by microbes has important consequences for microbiome composition and host health. Pigs do not encode all the enzymes needed to breakdown dietary polysaccharides, but bacteria contain numerous carbohydrate-active enzymes (CAZys) which contribute to this task. In addition, microbial fermentation of carbohydrates produces many bioactive metabolites such as short chain fatty acids (SCFAs). The relationships between microbes, dietary components, and the pig are complex, but a thorough understanding of these interactions would promote the design of interventions which target the microbiome and improve swine growth, especially during the critical weaning transition. At weaning, the microbiome undergoes rapid compositional changes as the diet shifts from milk to plant based. Here, we use quantitative (TMT)-based liquid chromatography tandem mass spectrometry (LC-MS/MS) to identify and quantitate piglet and microbial proteins in the gut during the weaning transition, with a focus on those involved in carbohydrate breakdown and nutrient synthesis. We analyzed 6 fecal samples from healthy, nursery piglets at weaning [day 21 postnatal (PN)] and 6 samples taken two weeks later, after acclimation to nursery diet (day 49 PN). In total, we detected 14,319 bacterial and fungal protein groups and 1,835 pig protein groups, and abundances of roughly one-half of the protein groups changed significantly over time (P = 0.05, fold change &amp;gt; 1.5). As expected, from weaning to day 49 PN, expression of glycoside hydrolases (GHs) which degrade milk oligosaccharides decreased, while the abundance of most enzymes which degrade plant carbohydrates such as xylan, cellulose, and starch, increased. Some plant-degrading enzymes were most abundant during weaning, which may reflect the tendency for piglets to steal solid feed from their mothers during nursing. Subdoligranulum was one of the top producers of multiple GHs capable of degrading milk or plant carbohydrates, while other genera specialized in the production of enzymes which targeted either milk oligosaccharides (Blautia, Lachnoclostridium Clostridium, Fournierella, Faecalibacterium, Eisenbergiella) or plant carbohydrates (Prevotella, Selemonas, Anaerovibrio). Aspergillus produced an uncharacterized protein predicted to degrade plant xyloglucan, highlighting the contributions of fungi in swine digestion. Host glycan-degrading enzymes decreased over time. Last, we detected acetate and butyrate production enzymes including acetate kinase (26 genera) and phosphate acetyltransferase (9 genera), and butyrate kinase and phosphate butyryltransferase from Subdoligranulum. Together, these data identify specific enzymes and the microbes that produce them during the transition from a milk to plant-based diet. This information provides a more robust understanding of digestion in the weanling pig and will ultimately provide a basis for effective design of probiotics and other therapeutics which target the gut microbiome to enhance swine growth and productivity.

Tissue-specific transcriptional response of post-larval clownfish to ocean warming

Anthropogenically driven climate change is predicted to increase average sea surface temperatures, as well as the frequency and intensity of marine heatwaves in the future. This increasing temperature is predicted to have a range of negative physiological impacts on multiple life-stages of coral reef fish. Nevertheless, studies of early-life stages remain limited, and tissue-specific transcriptomic studies of post-larval coral reef fish are yet to be conducted. Here, in an aquaria-based study we investigate the tissue-specific (brain, liver, muscle, and digestive tract) transcriptomic response of post-larval (20 dph) Amphiprion ocellaris to temperatures associated with future climate change (+3 °C). Additionally, we utilized metatranscriptomic sequencing to investigate how the microbiome of the digestive tract changes at +3 °C. Our results show that the transcriptional response to elevated temperatures is highly tissue-specific, as the number of differentially expressed genes (DEGs) and gene functions varied amongst the brain (102), liver (1785), digestive tract (380), and muscle (447). All tissues displayed DEGs associated with thermal stress, as 23 heat-shock protein genes were upregulated in all tissues. Our results indicate that post-larval clownfish may experience liver fibrosis-like symptoms at +3 °C as genes associated with extracellular matrix structure, oxidative stress, inflammation, glucose transport, and metabolism were all upregulated. We also observe a shift in the digestive tract microbiome community structure, as Vibrio sp. replace Escherichia coli as the dominant bacteria. This shift is coupled with the dysregulation of various genes involved in immune response in the digestive tract. Overall, this study highlights post-larval clownfish will display tissue-specific transcriptomic responses to future increases in temperature, with many potentially harmful pathways activated at +3 °C.

Urinary microbiome community types associated with urinary incontinence severity in women

BackgroundUrinary microbiome (urobiome) studies have previously reported on specific taxa and community differences in women with mixed urinary incontinence compared to controls. Therefore, we hypothesized that higher urinary and vaginal microbiome diversity would be associated with increased urinary incontinence severity. ObjectivesTo test if specific urinary or vaginal microbiome community-types are associated with urinary incontinence severity in a population of women with mixed urinary incontinence. Study DesignThis planned secondary, cross-sectional analysis evaluated associations between the urinary/vaginal microbiomes and urinary incontinence severity in a subset of ESTEEM trial participants with urinary incontinence. Incontinence severity was measured using bladder diaries and Urinary Distress Inventory (UDI) questionnaires collected at baseline. Catheterized urine samples and vaginal swabs were concurrently collected prior to treatment at baseline to assess the urinary and vaginal microbiomes.16S rRNA V4-V6 variable regions were sequenced, characterizing bacterial taxa to the genus level using the DADA2 pipeline and SILVA database. Using Dirichlet multinomial mixture methods, samples were clustered into community-types based on core taxa. Associations between community-types and severity measures (UDI total, UDI subscale scores, and the number of urinary incontinence episodes (total, urgency, and stress) from bladder diary) were evaluated using linear regression models adjusted for age and body mass index. Alpha diversity measures for richness (total taxa numbers) and evenness (proportional distribution of taxa abundance) were also analyzed for associations with urinary incontinence episodes and community-type. ResultsSix urinary microbiome community-types were identified, characterized by varying levels of common genera (Lactobacillus, Gardnerella, Prevotella, Tepidomonas, Acidovorax, Escherichia, and others). Analysis of urinary incontinence severity in 126 participants with MUI identified a Lactobacillus-dominated reference group with the highest abundance of Lactobacillus (76% mean relative abundance). A community characterized by fewer Lactobacilli (19% mean relative abundance) and greater alpha diversity, was associated with higher total urinary incontinence episodes (2.67 daily leaks, 95% CI: 0.76 - 4.59, p=0.007) and urgency urinary incontinence episodes (1.75 daily leaks, 95% CI: 0.24 - 3.27, p=0.02) compared to the reference group. No significant association was observed between community-type and stress urinary incontinence episodes or UDI total or subscores. The composition of vaginal community-types and urinary community-types were similar but comprised of slightly different bacterial taxa. Vaginal community-types were not associated with urinary incontinence severity, as measured by bladder diary or UDI total and subscale scores. Alpha diversity indicated that greater sample richness was associated with more incontinence episodes (observed genera p=0.01) in urine. Measures of evenness (Shannon/Pielou) were not associated with incontinence severity in the urinary or vaginal microbiomes. ConclusionsIn the urobiome of women with mixed urinary incontinence, a community-type with fewer Lactobacilli and more diverse bacteria was associated with more severe urinary incontinence episodes (total and urgency) compared to a community-type with high predominance of a single genus, Lactobacillus. Whether mixed urinary incontinence severity is due to lesser predominance of Lactobacillus, greater presence of other non-Lactobacillus genera, or the complement of bacteria comprising urobiome community-types, remains to be determined.

Effect of structural characteristics of resistant starch prepared by various methods on microbial community and fermentative products

Resistant starch (RS) has been extensively studied because of its beneficial effects on gut microbiota. In this study, four RSs obtained through various preparation processes were utilized for in vitro fermentation, and their structural characteristics before and after fermentation were determined using chromatography, Fourier infrared spectroscopy, and scanning electron microscopy (SEM). It was observed that these RSs can be classified into two categories based on their fermentation and structural features. The autoclaving RS (ARS) and extruding RS (ERS) were classified as Class I Microbiome Community (MC-I), characterized by a higher proportion of butyrate and its producers, including unclassified_g_Megasphaera and Megasphaera elsdenii. While microwaving RS (MRS) and ultrasound RS (URS) belonged to Class II Microbiome Community (MC-II), marked by a higher proportion of acetate and its producer, Bifidobacterium pseudocatenulatum DSM 20438. MC-I had a lower molecular weight, shorter chain length, more chains with degree of polymerization (DP) 36–100, and a more ordered structure than MC-II. Furthermore, SEM observations revealed distinct degradation patterns between MC-I and MC-II, which may be attributed to their surface structural characteristics. These findings imply that the preparation methods employed for RS can determine its multilevel structural characteristics, and consequently influence its physiological properties.

Influence of organic plant breeding on the rhizosphere microbiome of common bean (Phaseolus vulgaris L.).

We now recognize that plant genotype affects the assembly of its microbiome, which in turn, affects essential plant functions. The production system for crop plants also influences the microbiome composition, and as a result, we would expect to find differences between conventional and organic production systems. Plant genotypes selected in an organic regime may host different microbiome assemblages than those selected in conventional environments. We aimed to address these questions using recombinant inbred populations of snap bean that differed in breeding history. Rhizosphere microbiomes of conventional and organic common beans (Phaseolus vulgaris L.) were characterized within a long-term organic research site. The fungal and bacterial communities were distinguished using pooled replications of 16S and ITS amplicon sequences, which originated from rhizosphere samples collected between flowering and pod set. Bacterial communities significantly varied between organic and conventional breeding histories, while fungal communities varied between breeding histories and parentage. Within the organically-bred populations, a higher abundance of a plant-growth-promoting bacteria, Arthrobacter pokkalii, was identified. Conventionally-bred beans hosted a higher abundance of nitrogen-fixing bacteria that normally do not form functional nodules with common beans. Fungal communities in the organically derived beans included more arbuscular mycorrhizae, as well as several plant pathogens. The results confirm that the breeding environment of crops can significantly alter the microbiome community composition of progeny. Characterizing changes in microbiome communities and the plant genes instrumental to these changes will provide essential information about how future breeding efforts may pursue microbiome manipulation.

Food desert residence has limited impact on veteran fecal microbiome composition: a U.S. Veteran Microbiome Project study.

Social and economic inequities can have a profound impact on human health. The inequities could result in alterations to the gut microbiome, an important factor that may have profound abilities to alter health outcomes. Moreover, the strong correlations between social and economic inequities have been long understood. However, to date, limited research regarding the microbiome and mental health within the context of socioeconomic inequities exists. One particular inequity that may influence both mental health and the gut microbiome is living in a food desert. Persons living in food deserts may lack access to sufficient and/or nutritious food and often experience other inequities, such as increased exposure to air pollution and poor access to healthcare. Together, these factors may confer a unique risk for microbial perturbation. Indeed, external factors beyond a food desert might compound over time to have a lasting effect on an individual's gut microbiome. Therefore, adoption of a life-course approach is expected to increase the ecological validity of research related to social inequities, the gut microbiome, and physical and mental health.

Metal transporter SLC39A14/ZIP14 modulates regulation between the gut microbiome and host metabolism.

Metal transporter SLC39A14/ZIP14 is localized on the basolateral side of the intestine, functioning to transport metals from blood to intestine epithelial cells. Deletion of Slc39a14/Zip14 causes spontaneous intestinal permeability with low-grade chronic inflammation, mild hyperinsulinemia, and greater body fat with insulin resistance in adipose. Importantly, antibiotic treatment reverses the adipocyte phenotype of Slc39a14/Zip14 knockout (KO), suggesting a potential gut microbial role in the metabolic alterations in the Slc39a14/Zip14 KO mice. Here, we investigated the hypothesis that increased intestinal permeability and subsequent metabolic alterations in the absence of Zip14 could be in part due to alterations in gut microbial composition. Dietary metals have been shown to be involved in the regulation of gut microbial diversity and composition. However, studies linking the action of intestinal metal transporters to gut microbial regulation are lacking. We showed the influence of deletion of metal transporter Slc39a14/Zip14 on gut microbiome composition and how ZIP14-linked changes to gut microbiome community composition are correlated with changes in host metabolism. Deletion of Slc39a14/Zip14 generated Zn-deficient epithelial cells and luminal content in the entire intestinal tract, a shift in gut microbial composition that partially overlapped with changes previously associated with obesity and inflammatory bowel disease (IBD), increased the fungi/bacteria ratio in the gut microbiome, altered the host metabolome, and shifted host energy metabolism toward glucose utilization. Collectively, our data suggest a potential predisease microbial susceptibility state dependent on host gene Slc39a14/Zip14 that contributes to intestinal permeability, a common trait of IBD, and metabolic disorders such as obesity and type 2 diabetes.NEW & NOTEWORTHY Metal dyshomeostasis, intestinal permeability, and gut dysbiosis are emerging signatures of chronic disorders, including inflammatory bowel diseases, type-2 diabetes, and obesity. Studies in reciprocal regulations between host intestinal metal transporters genes and gut microbiome are scarce. Our research revealed a potential predisease microbial susceptibility state dependent on the host metal transporter gene, Slc39a14/Zip14, that contributes to intestinal permeability providing new insight into understanding host metal transporter gene-microbiome interactions in developing chronic disease.

Improved phyllosphere microbiome composition of tea plant with the application of small peptides in combination with rhamnolipid

BackgroundSmall peptides play a crucial role in plant growth and adaptation to the environment. Exogenous small peptides are often applied together with surfactants as foliar fertilizers, but the impact of small peptides and surfactants on the tea phyllosphere microbiome remains unknown.ResultsIn this study, we investigated the effects of small peptides and different surfactants on the tea phyllosphere microbiome using 16S and ITS sequencing. Our results showed that the use of small peptides reduced the bacterial diversity of the tea phyllosphere microbiome and increased the fungal diversity, while the use of surfactants influenced the diversity of bacteria and fungi. Furthermore, the addition of rhamnolipid to small peptides significantly improved the tea phyllosphere microbiome community structure, making beneficial microorganisms such as Pseudomonas, Chryseobacterium, Meyerozyma, and Vishniacozyma dominant populations.ConclusionOur study suggests that the combined use of small peptides and surfactants can significantly modify the tea phyllosphere microbiome community structure, particularly for beneficial microorganisms closely related to tea plant health. Thus, this preliminary study offers initial insights that could guide the application of small peptides and surfactants in agricultural production, particularly with respect to their potential for modulating the phyllosphere microbiome community in tea plant management.

Sediment microbial community characteristics in sea cucumber restocking area

Variations of microbial species and functional composition in coastal sediment are usually taken as the results of the provision of supplementary nutrients affected by human activities. However, responses of microbiome stability to restocking biological resources remain less understood in coastal benthic systems without nutrient supplements. Here, combined with metagenomics and microbiome co-occurrence networks, the composition, function, and community stability of microbes were evaluated in a coastal area where sea cucumbers (Apostichopus japonicus) restocked after six months. Also, the physicochemical characteristics of sediments and bottom water were analyzed. We found the total organic carbon, total nitrogen, and total phosphorus of sediment did not change significantly in the restocking area after six months, whereas the concentration of dissolved inorganic nitrogen in bottom water increased significantly. Moreover, the relative abundance of Nitrospina at the class level was increased significantly in the restocking area. Also, enzymes related to nitrate reduction and nitrous oxide reductase were increased in the restocking area. Of note, stock enhancement of sea cucumbers altered associations between bacteria rather than their composition. The elimination of negative associations and reduction of the potential keystone taxa in the restocking area indicated destabilized bacterial communities. Our work may contribute to elucidating the response of microbial stability to stock enhancement. This finding also suggests that microbial community stability can be considered as an indicator of ecological risk under the influence of stock enhancement.

Chronic exposure to pesticides disrupts the bacterial and fungal co-existence and the cross-kingdom network characteristics of honey bee gut microbiome

Gut microbiome communities have a significant impact on bee health and disease and have been shown to be shaped by a variety of factors, including exposure to pesticides and inhive chemicals. However, it is unknown whether pesticide exposure affects the coexistence and cross-kingdom network parameters of bee gut microbiome communities because microbes may compete in the gut environment under different stressors. Therefore, we conducted additional analysis of the microbiome data from our previous study in which we discovered that exposure to two novel insecticides flupyradifurone (FPF) and sulfoxaflor (Sulf) or/and a fungicide, azoxystrobin (Azoxy) caused dysbiosis of bee gut microbiota that was associated with an increase in the relative abundance of opportunistic pathogens such as Serratia marcescens. We investigated for the first time the potential cross-kingdom fungal-bacterial interactions using co-occurrence pattern correlation and network analysis. We discovered that exposure to FPF or Sulf alone or in combination with Azoxy fungicide influenced the co-existence patterns of fungal and bacterial communities. Significant differences in degree centrality, closeness centrality, and eigenvector centrality distribution indices were also found in single and double-treatment groups compared to controls. The effects of FPF and Sulf alone on cross-kingdom parameters (bacterial to fungal node ratio, degree of centrality, closeness centrality, and eigenvector centrality) were distinct, but this was reversed when they were combined with Azoxy fungicide. The fungal and bacterial hub taxa identified differed, with only a few shared hubs across treatments, suggesting microbial cross-kingdom networks may be disrupted differently under different stressors. Our findings add to our understanding of pesticide effects on the bee gut microbiome and bee health in general, while also emphasizing the importance of cross-kingdom network analysis in future microbiome research.

Microbial signatures of vertebrate visitation in floral nectar: a case study with two endemic Aotearoa New Zealand plant species

ABSTRACT Microbes found in nectar and dispersed by animal visitors can mediate pollination and host fitness. While previous studies have characterised floral nectar microbiomes and their effects on invertebrate pollinators, fewer studies have focused on vertebrate pollinators, especially outside of the Northern Hemisphere. In Aotearoa New Zealand, vertebrates, such as birds and bats, are critical pollinators for many native plant species. Here, we present nectar microbiome profiles for two endemic, vertebrate-pollinated plant species in Aotearoa New Zealand, wharariki, mountain flax (Phormium cookianum), and pua o te Rēinga, wood rose (Dactylanthus taylorii). We used vertebrate exclusion treatments and camera traps to monitor visitation. Camera trap footage revealed silvereye (Zosterops lateralis), ship rat (Rattus rattus), and possum (Trichosurus vulpecula) visitation. We detected shifts in microbial species turnover in wood rose and varying relative abundances of fungal and bacterial taxa across the vertebrate exclusion treatments for both mountain flax and wood rose. However, we did not detect strong effects on floral nectar microbiome community composition or richness. Future work should move beyond profiling the microbial communities and identify fitness consequences and pollination outcomes, which could affect conservation and management decisions.

Nisin a probiotic bacteriocin mitigates brain microbiome dysbiosis and Alzheimer’s disease-like neuroinflammation triggered by periodontal disease

IntroductionPeriodontitis-related oral microbial dysbiosis is thought to contribute to Alzheimer's disease (AD) neuroinflammation and brain amyloid production. Since probiotics can modulate periodontitis/oral dysbiosis, this study examined the effects of a probiotic/lantibiotic, nisin, in modulating brain pathology triggered by periodontitis.MethodsA polymicrobial mouse model of periodontal disease was used to evaluate the effects of this disease on brain microbiome dysbiosis, neuroinflammation, Alzheimer’s-related changes, and nisin’s therapeutic potential in this context.Results16S sequencing and real-time PCR data revealed that Nisin treatment mitigated the changes in the brain microbiome composition, diversity, and community structure, and reduced the levels of periodontal pathogen DNA in the brain induced by periodontal disease. Nisin treatment significantly decreased the mRNA expression of pro-inflammatory cytokines (Interleukin-1β/IL-1 β, Interleukin 6/IL-6, and Tumor Necrosis Factor α/TNF-α) in the brain that were elevated by periodontal infection. In addition, the concentrations of amyloid-β 42 (Aβ42), total Tau, and Tau (pS199) (445.69 ± 120.03, 1420.85 ± 331.40, 137.20 ± 36.01) were significantly higher in the infection group compared to the control group (193.01 ± 31.82, 384.27 ± 363.93, 6.09 ± 10.85), respectively. Nisin treatment markedly reduced the Aβ42 (261.80 ± 52.50), total Tau (865.37 ± 304.93), and phosphorylated Tau (82.53 ± 15.77) deposition in the brain of the infection group.DiscussionNisin abrogation of brain microbiome dysbiosis induces beneficial effects on AD-like pathogenic changes and neuroinflammation, and thereby may serve as a potential therapeutic for periodontal–dysbiosis-related AD.

In Vitro Dental Plaque Culture Model for Biofilm Structural Analyses.

Extensive research has focused on the compositional changes in dental plaque microbiome communities during the transition from health to disease, known as dysbiosis. However, alterations in the spatial composition of these communities throughout the progression from health to disease remain under-explored. We describe an in vitro dental plaque model for culturing oral biofilms seeded with dental plaque from human volunteers. Our model recapitulates important features of the in vivo environment including shear force induced by salivary flow over teeth and the nutritional milieu experienced by microbes that inhabit the transitional zone between supragingival and subgingival aspects of the teeth. Importantly, our model is amenable to multiplex fluorescent labeling and multispectral imaging for testing specific hypotheses regarding systems-level community structure and function. The model allows for precise manipulation of various environmental conditions, such as flow rate and nutrient availability to investigate their effects on biofilm development and spatial structure. Furthermore, this model can be used to test the effects of various therapeutic interventions, e.g., antimicrobial agents, on the biofilm composition and structure at the micron to millimeter scale, making it a valuable tool for studying the molecular and cellular basis of dental plaque-mediated diseases and for benchmarking new therapeutic interventions. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Dental plaque-on-a-chip in vitro model culture system Support Protocol: Gingival margin (GM) medium preparation Basic Protocol 2: Microcosm labeling and multispectral image acquisition.

Characteristics of gut microbiome communities in the invasive African giant snail under urbanization gradient

To investigate the diversity and community structure of gut microbiome of the invasive species, Achatina fulica, along an urbanization gradient, we collected 30 A. fulica samples from five parks in the urban, suburban, and rural areas of Xiamen City. Using full-length 16S rRNA gene sequencing performed by the third generation PacBio sequencing platform, we analyzed the community characteristics of gut microbiome and soil microbiome in different habitats. We found a significant disparity between the composition of gut microbiome of A. fulica and that of the soil microbiome in their habitats. Furthermore, the gut microbiome of A. fulica were more sensitive to urbanization. The microbial α-diversity indices (Sobs, Chao, Shannon indices) in the soil of A. fulica habitats were consistently higher than those within their guts. Despite the similar β-diversity indices of microbial communities in urban, suburban, and rural soils, we found a significant discrepancy in gut microbiome composition. Urbanization significantly influenced A. fulica gut microbiome composition. Gut microbiome of A. fulica in urban and suburban regions primarily consisted of Enterobacteriaceae, Xanthomonadaceae, and Mycoplasmataceae, while that in rural areas chiefly composed of Streptococcaceae and Paenibacillaceae. The diversity and abundance of potential human pathogenic bacteria within the gut microbiome of A. fulica significantly increased in urban environments, suggesting that urbanization escalated the risk of A. fulica transmitting potential pathogens.