Microbiome Analysis Research Articles

Comparative analysis of the human microbiome from four different regions of China and machine learning-based geographical inference.

The human microbiome, the community of microorganisms that reside on and inside the human body, is critically important for health and disease. However, it is influenced by various factors and may vary among individuals residing in distinct geographic regions. In this study, 220 samples, consisting of sterile swabs from palmar skin and oral and nasal cavities were collected from Chinese Han individuals living in Shanghai, Chifeng, Kunming, and Urumqi, representing the geographic regions of east, northeast, southwest, and northwest China. The full-length 16S rRNA gene of the microbiota in each sample was sequenced using the PacBio single-molecule real-time sequencing platform, followed by clustering the sequences into operational taxonomic units (OTUs). The analysis revealed significant differences in microbial communities among the four regions. Cutibacterium was the most abundant bacterium in palmar samples from Shanghai and Kunming, Psychrobacter in Chifeng samples, and Psychrobacillus in Urumqi samples. Additionally, Streptococcus and Staphylococcus were the dominant bacteria in the oral and nasal cavities. Individuals from the four regions could be distinguished and predicted based on a model constructed using the random forest algorithm, with the predictive effect of palmar microbiota being better than that of oral and nasal cavities. The prediction accuracy using hypervariable regions (V3-V4 and V4-V5) was comparable with that of using the entire 16S rRNA. Overall, our study highlights the distinctiveness of the human microbiome in individuals living in these four regions. Furthermore, the microbiome can serve as a biomarker for geographic origin inference, which has immense application value in forensic science.IMPORTANCEMicrobial communities in human hosts play a significant role in health and disease, varying in species, quantity, and composition due to factors such as gender, ethnicity, health status, lifestyle, and living environment. The characteristics of microbial composition at various body sites of individuals from different regions remain largely unexplored. This study utilized single-molecule real-time sequencing technology to detect the entire 16S rRNA gene of bacteria residing in the palmar skin, oral, and nasal cavities of Han individuals from four regions in China. The composition and structure of the bacteria at these three body sites were well characterized and found to differ regionally. The results elucidate the differences in bacterial communities colonizing these body sites across different regions and reveal the influence of geographical factors on human bacteria. These findings not only contribute to a deeper understanding of the diversity and geographical distribution of human bacteria but also enrich the microbiome data of the Asian population for further studies.

Effect of pre-farrowing hygiene routine (sub-standard vs. optimal) and creep feeding regime (dry pelleted starter diet vs. liquid mixture of milk replacer and starter diet) on post-weaning intestinal parameters and growth to slaughter in pigs.

The objective was to evaluate the effect of providing dry pelleted starter diet (DPS) or a liquid mixture of milk replacer and starter diet (LMR+S) to suckling pigs housed in farrowing pens of sub-standard or optimal hygiene conditions on pig growth to slaughter, and post-weaning (PW) intestinal parameters. On day (d) 107 of gestation, 87 sows were randomly allocated to one of four treatments in a 2×2 factorial arrangement. The factors were creep feeding (DPS or LMR+S) and pre-farrowing hygiene routine (SUB-STANDARD or OPTIMAL). Pigs were provided with DPS (manually) from d11 to weaning (at d28±1.2 of age) or LMR+S using an automatic liquid feeding system from d4 to weaning. The SUB-STANDARD hygiene routine (pens washed and dried for ~18h, sows not washed or disinfected) and the OPTIMAL hygiene routine (pens pre-soaked, detergent applied, washed, dried for 3 days, chlorocresol-based disinfectant applied, dried for 3 more days and sows washed and disinfected with Virkon) were used to obtain SUB-STANDARD or OPTIMAL hygiene conditions, respectively in farrowing rooms prior to entry of sows. Microbiome analysis was performed on fecal samples from 8 focal pigs/treatment, before weaning and at d21 and d114 PW. On d4 PW, 10 pigs/treatment were euthanized to collect intestinal tissue and digesta samples for histological, enzyme activity and microbiome analysis. Feeding LMR+S to pigs born into the OPTIMAL hygiene increased total dry matter intake compared to all of the other groups (P ≤ 0.05) and increased weaning weight compared to DPS feeding under both OPTIMAL and SUB-STANDARD hygiene conditions (P ≤ 0.05). Pigs from OPTIMAL farrowing pens had lower clinical cases of disease, diarrhoea prevalence and were slaughtered 3.8 days earlier than those from SUB-STANDARD farrowing pens (P ≤ 0.05). Suckling piglet mortality was reduced with LMR+S (P ≤ 0.05). On d4 PW, jejunal and ileal villus height were increased by OPTIMAL hygiene and ileal sucrase activity was increased by LMR+S (P ≤ 0.05). On d4 PW, LMR+S-fed pigs from OPTIMAL farrowing pens had lower relative abundance of Clostridium_P in the jejunum. In conclusion, the OPTIMAL hygiene routine increased pre-weaning LMR+S feed intake, reduced clinical cases of disease, improved intestinal structure and reduced the weaning to slaughter duration, while LMR+S feeding increased weaning weight, intestinal maturity and reduced pre-weaning mortality.

Herbicide-treated soil as a reservoir of beneficial bacteria: microbiome analysis and PGP bioinoculants in maize

BackgroundHerbicides are integral to agricultural weed management but can adversely affect non-target organisms, soil health, and microbiome. We investigated the effects of herbicides on the total soil bacterial community composition using 16S rRNA gene amplicon community profiling. Further, we aimed to identify herbicide-tolerant bacteria with plant growth-promoting (PGP) capabilities as a mitigative strategy for these negative effects, thereby promoting sustainable agricultural practices.ResultsA bacterial community analysis explored the effects of long-term S-metolachlor application on soil bacterial diversity, revealing that the herbicide’s impact on microbial communities is less significant than the effects of temporal factors (summer vs. winter) or agricultural practices (continuous maize cultivation vs. maize-winter wheat rotation). Although S-metolachlor did not markedly alter the overall bacteriome structure in our environmental context, the application of enrichment techniques enabled the selection of genera such as Pseudomonas, Serratia, and Brucella, which were rare in metagenome analysis of soil samples. Strain isolation revealed a rich source of herbicide-tolerant PGP bacteria within the culturable microbiome fraction, termed the high herbicide concentration tolerant (HHCT) bacterial culture collection.Within the HHCT collection, we isolated 120 strains that demonstrated significant in vitro PGP and biocontrol potential, and soil quality improvement abilities. The most promising HHCT isolates were combined into three consortia, each exhibiting a comprehensive range of plant-beneficial traits. We evaluated the efficacy and persistence of these multi-strain consortia during 4-week in pot experiments on maize using both agronomic parameters and 16S rRNA gene community analysis assessing early-stage plant development, root colonization, and rhizosphere persistence. Notably, 7 out of 10 inoculated consortia partners successfully established themselves and persisted in the maize root microbiome without significantly altering host root biodiversity. Our results further evidenced that all three consortia positively impacted both seed germination and early-stage plant development, increasing shoot biomass by up to 47%.ConclusionsHerbicide-treated soil bacterial community analysis revealed that integrative agricultural practices can suppress the effects of continuous S-metolachlor application on soil microbial diversity and stabilize microbiome fluctuations. The HHCT bacterial collection holds promise as a source of beneficial bacteria that promote plant fitness while maintaining herbicide tolerance.

Alterations in Gut Microbiota Composition Are Associated with Changes in Emotional Distress in Children with Obstructive Sleep Apnea

Emerging evidence underscores the pivotal role of the gut microbiota in regulating emotional and behavioral responses via the microbiota–gut–brain axis. This study explores associations between pediatric obstructive sleep apnea (OSA), emotional distress (ED), and gut microbiome alterations before and after OSA treatment. Sixty-six children diagnosed with OSA via polysomnography participated, undergoing adenotonsillectomy alongside routine educational sessions. ED was assessed using the OSA-18 questionnaire, categorizing participants into high ED (scores ≥ 11, 52%) and low ED (scores < 11, 48%) groups. Gut microbiome analysis revealed significant diversity differences, with high ED linked to a reduced Shannon index (p = 0.03) and increased beta diversity (p = 0.01). Three months post-treatment, significant improvements were observed in OSA symptoms, ED scores, and gut microbiome alpha diversity metrics among 55 participants (all p < 0.04). Moreover, changes in the relative abundances of Veillonella, Bifidobacterium, Flavonifractor, and Agathobacter, as well as ultra-low frequency power and low frequency power of sleep heart rate variability, were independently associated with ED score alterations. These findings underscore the gut microbiome’s critical role in the emotional and behavioral symptoms associated with pediatric OSA, suggesting that microbiome-targeted interventions could complement traditional treatments for ED reduction and emphasizing the need for further research.

Millennial-scale microbiome analysis reveals ancient antimicrobial resistance conserved despite modern selection pressures

BackgroundAntimicrobial resistance presents a formidable challenge, yet its existence predates the introduction of antibiotics. Our study delves into the presence of antimicrobial resistance genes (ARGs) in ancient permafrost microbiomes, comparing them with contemporary soil and pristine environments. Majority of the samples are from regions around Beringia, encompassing parts of Russia and Alaska, with only one sample originating from the Tien Shan Mountain range in Kyrgyzstan.ResultsFrom over 2.3 tera basepairs of raw metagenomic data, retrieved from samples ranging in age from approximately 7,000 years to 1.1 million years, we assembled about 1.3 billion metagenomic contigs and explored the prevalence of ARGs within them. Our findings reveal a diverse array of ARGs in ancient microbiomes, akin to contemporary counterparts. On average, we identified 2 ARGs per rRNA gene in ancient samples. Actinomycetota, Bacillota, and several thermophiles were prominent carriers of ARGs in Chukochi and Kamchatkan samples. Conversely, ancient permafrost from the Tien Shan Mountain range exhibited no Thermophiles or Actinomycetota carrying ARGs. Both ancient and contemporary microbiomes showcased numerous divergent ARGs, majority of which have identity between 40 and 60% to genes in antibiotic resistance gene databases. To study the selection pressure on ARGs, we performed dN/dS analysis specifically on antibiotic inactivation-type ARGs, which exhibited purifying selection compared to contemporary genes.ConclusionAntibiotic resistance has existed throughout microbial evolution and will likely persist, as microbes have the capacity to develop and retain resistance genes through evolutionary processes. The classes of antimicrobial resistance genes profiled and the function of antibiotic-inactivating enzymes from ancient permafrost microbiomes do not seem to be very different from the genes found in the antibiotic era. Additionally, we retrieved 359 putative complete viruses from ancient microbiomes and none of them harboured any ARGs.

Metabarcoding analysis of oral microbiome during pregnancy

Pregnancy is a dynamic physiological process involving significant hormonal, immune, and metabolic changes to support fetal growth and development. This study investigates the changes in salivary microbiome and biochemical markers from the second to the third trimester of pregnancy. Saliva samples were collected from 45 pregnant women enrolled in the Qatar Birth Cohort study at two time points (second and third trimesters). DNA was extracted and subjected to 16S rRNA gene sequencing using Oxford Nanopore Technology. Microbial diversity and taxonomic analyses were performed, along with correlation analyses between microbial abundance and clinical parameters. Biochemically, significant increases in BMI, pulse rate, HbA1c, LDL, total cholesterol, and triglycerides were observed in the third trimester compared to the second. Microbial diversity analysis revealed significant changes in microbial richness and composition. Taxonomy analysis showed a significant 3-fold increase in Bacteroidota. Also, a significant decline in Selenomonas and a significant increase in Veillonella, specifically Veillonella dispar and Veillonella atypica, as well as an increase in Granulicatella were observed in the third trimester, along with a significant decrease in Streptococcus sanguinis. Correlation analysis during the second trimester revealed positive associations between BMI, cholesterol, LDL, and Selenomonas, and negative correlations with Streptococcus and Gemella. In the third trimester, BMI was negatively correlated with Campylobacter, glucose levels were negatively correlated with Neisseria, and triglyceride levels were negatively correlated with Prevotella. These findings highlight significant biochemical and microbial shifts during pregnancy, underscoring the importance of monitoring oral health and metabolic changes in pregnant women.

Dietary prevention of antibiotic-induced dysbiosis and mortality upon aging in mice.

Oral antibiotic use is both widespread and frequent in older adults and has been linked to dysbiosis of the gut microbiota, enteric infection, and chronic diseases. Diet and nutrients, particularly prebiotics, may modify the susceptibility of the gut microbiome to antibiotic-induced dysbiosis. We fed 12-month-old mice a high glycemic (HG) or low glycemic (LG) diet with or without antibiotics (ampicillin and neomycin) for an additional 11 months. The glycemic index was modulated by the ratio of rapidly digested amylopectin starch to slowly digested amylose, a type-2-resistant starch. We observed a significant decrease in survival of mice fed a HG diet containing antibiotics (HGAbx) relative to those fed a LG diet containing antibiotics (LGAbx). HGAbx mice died with an enlarged and hemorrhagic cecum, which is associated with colonic hyperplasia and goblet cell depletion. Gut microbiome analysis revealed a pronounced expansion of Proteobacteria and a near-complete loss of Bacteroidota and Firmicutes commensal bacteria in HGAbx, whereas the LGAbx group maintained a population of Bacteroides and more closely resembled the LG microbiome. The predicted functional capacity for bile salt hydrolase activity was lost in HGAbx mice but retained in LGAbx mice. An LG diet containing amylose may therefore be a potential therapeutic to prevent antibiotic-induced dysbiosis and morbidity.

Human oral microbiome as forensic biomarkers for individual identification: A systematic review

: The oral microbiome is a promising and novel source of biomarkers for forensic identification. It offers distinct signatures that may differentiate individuals, sparking new avenues of research and discovery.: This systematic review is meticulously designed to comprehensively consolidate current research on oral microbiome signatures in forensic contexts, leaving no stone unturned in our quest for knowledge. It focuses explicitly on microbial diversity, community structure, and specific taxa as potential markers for individual identification.s: To identify relevant studies published between 2014 and 2024, a comprehensive search of major scientific databases was conducted. Inclusion criteria encompassed peer-reviewed articles investigating oral microbiome diversity and specific microbial markers relevant to forensic individual identification. These studies were employed to evaluate the reliability and consistency of these biomarkers across diverse populations and environmental conditions.: The review included 13 studies that met the criteria, revealing significant associations between specific oral microbial taxa and individual identification. This systematic review highlighted consistent discriminatory power and stability of microbial signatures across varied populations.: Oral microbiome analysis is promising to enhance forensic investigations by providing unique biomarkers for individual identification. However, challenges such as standardization of microbiome studies and influences of environmental factors on microbiome underscore the need for further research to validate and effectively implement oral microbiome data in forensic practice.

Phylosymbiosis and Elevated Cancer Risk in Genetically Depauperate Channel Island Foxes.

Examination of the host-associated microbiome in wildlife can provide critical insights into the eco-evolutionary factors driving species diversification and response to disease. This is particularly relevant for isolated populations lacking genomic variation, a phenomenon that is increasingly common as human activities create habitat 'islands' for wildlife. Here, we characterised the gut and otic microbial communities of one such species: Channel Island foxes (Urocyon littoralis). The gut microbiome provided evidence of phylosymbiosis by reflecting the host phylogeny, geographic proximity, history of island colonisation and contemporary ecological differences, whereas the otic microbiome primarily reflected geography and disease. Santa Catalina Island foxes are uniquely predisposed to ceruminous gland tumours following infection with Otodectes cynotis ear mites, while San Clemente and San Nicolas Island foxes exhibit ear mite infections without evidence of tumours. Comparative analyses of otic microbiomes revealed that mite-infected Santa Catalina and San Clemente Island foxes exhibited reduced bacterial diversity, skewed abundance towards the opportunistic pathogen Staphylococcus pseudintermedius and disrupted microbial community networks. However, Santa Catalina Island foxes uniquely harboured Fusobacterium and Prevotella bacteria as potential keystone taxa. These bacteria have previously been associated with colorectal cancer and may predispose Santa Catalina Island foxes to an elevated cancer risk. In contrast, mite-infected San Nicolas Island foxes maintained high bacterial diversity and robust microbial community networks, suggesting that they harbour more resilient microbiomes. Considered together, our results highlight the diverse eco-evolutionary factors influencing commensal microbial communities and their hosts and underscore how the microbiome can contribute to disease outcomes.

Two rice cultivars recruit different rhizospheric bacteria to promote aboveground regrowth after mechanical defoliation.

Plants have evolved the ability to regrow after mechanical defoliation and environmental stresses. However, it is unclear whether and how defoliated plants exploit beneficial microbiota from the soil to promote aboveground regrowth. Here, we compared the defoliation-triggered changes in the root exudation and bacterial microbiome of two rice cultivars (Oryza sativa L ssp.), indica/xian cultivar Minghui63 and japonica/geng cultivar Nipponbare. The results show that reciprocal growth promotion existed between defoliated Minghui63 seedlings and soil bacteria. After the leaves were removed, the Minghui63 seedlings displayed approximately 1.5- and 2.1-fold higher root exudation and leaf regrowth rates, respectively, than did the Nipponbare seedlings. In field trials, Minghui63 and Nipponbare enriched taxonomically and functionally distinct bacteria in the rhizosphere and root. In particular, Minghui63 rhizosphere and root communities depleted bacteria whose functions are related to xenobiotics biodegradation and metabolism. The microbiome data implied that the bacterial family Rhodocyclaceae was specifically enriched during the regrowth of defoliated Minghui63 rice. We further isolated a Rhodocyclaceae strain, Uliginosibacterium gangwonense MDD1, from rice root. Compared with germ-free conditions, MDD1 inoculation promoted the aboveground regrowth of defoliated Minghui63 by 61% but had a weaker effect on Nipponbare plants, suggesting cultivar-specific associations between regrowth-promoting bacteria and rice. This study provides novel insight into microbiota‒root‒shoot communication, which is implicated in the belowground microbiome and aboveground regrowth in defoliated rice. These data will be helpful for microbiome engineering to increase rice resilience to defoliation and environmental stresses.IMPORTANCEAs sessile organisms, plants face a multitude of abiotic and biotic stresses which often result in defoliation. To survive, plants have evolved the ability to regrow leaves after stresses and wounding. Previous studies revealed that the rhizosphere microbiome affected plant growth and stress resilience; however, how belowground microbiota modulates the aboveground shoot regrowth is unclear. To address this question, we used rice, an important crop worldwide, to analyze the role of rhizosphere microbiota in leaf regrowth after defoliation. Our data indicate mutual growth promotion between defoliated rice and rhizosphere bacteria and such beneficial effect is cultivar specific. The microbiome analysis also led us to find a Uliginosibacterium gangwonense strain that promoted rice cv. MH63 leaf regrowth. Our findings therefore present a novel insight into plant-microbiome function and provide beneficial strains that potentially enhance rice stress resilience.

Bibliometric and visual analysis of human microbiome—breast cancer interactions: current insights and future directions

The composition of the gut microbiome differs from that of healthy individuals and is closely linked to the progression and development of breast cancer. Recent studies have increasingly examined the relationship between microbial communities and breast cancer. This study analyzed the research landscape of microbiome and breast cancer, focusing on 736 qualified publications from the Web of Science Core Collection (WoSCC). Publications in this field are on the rise, with the United States leading in contributions, followed by China and Italy. Despite this strong output, the centrality value of China in this field is comparatively low at ninth, highlighting a gap between the quantity of research and its global impact. This pattern is repetitively observed in institutional contributions, with a predominance of Western institutes among the top contributors, underscoring a potential research quality gap in China. Keyword analysis reveals that research hotspots are focused on the effect of microbiome on breast cancer pathogenesis and tumor metabolism, with risk factors and metabolic pathways being the most interesting areas. Publications point to a shift toward anti-tumor therapies and personalized medicine, with clusters such as “anti-tumor” and “potential regulatory agent” gaining prominence. Additionally, intratumor bacteria studies have emerged as a new area of significant interest, reflecting a new direction in research. The University of Helsinki and Adlercreutz H are influential institutions and researchers in this field. Current trends in microbiome and breast cancer research indicate a significant shift toward therapeutic applications and personalized medicine. Strengthening international collaborations and focusing on research quality is crucial for advancing microbiome and breast cancer research.

Integrating microbial profiling and machine learning for inference of drowning sites: a forensic investigation in the Northwest River.

Drowning incidents present significant challenges for forensic investigators in determining the exact site of occurrence. Traditional forensic methods often rely on physical evidence and circumstantial clues, but the emerging field of forensic microbiology offers a promising avenue for enhancing precision and reliability in site inference. Our study investigates the application of microbiome analysis in inferring drowning sites, focusing on microbial diversity in water samples and lung tissues of drowned animals from different sites in the Northwest River. We utilized 16S rDNA sequencing to analyze microbial diversity in water samples and lung tissues, revealing distinct microbial signatures associated with drowning sites. Our findings highlight variations in species richness and diversity across different sampling points, indicating the influence of environmental factors on microbial community structure. Machine learning models trained on microbial data from lung tissues demonstrated high accuracy in predicting drowning sites, with cross-validation accuracy ranging from 83.53% ± 3.99% to 95.07% ± 3.17%. Notably, the Gradient Boosting Machine (GBM) method achieved a classification accuracy of 95.07% ± 3.17% for different sampling points at a submersion time of 1 day. Moreover, our cross-species site inference results revealed that utilizing data from drowned mice to predict the drowning sites of rabbits in location W5 achieved an accuracy of 72.22%. In conclusion, our study underscores the potential of microbiome analysis in forensic investigations of drowning incidents. By integrating microbial data with traditional forensic techniques, there is significant potential to enhance the reliability of scene inferences, thereby making substantial contributions to case investigations and judicial trials.IMPORTANCEBy employing advanced techniques like microbial profiling and machine learning, the study aims to enhance the accuracy of determining drowning sites, which is crucial for both legal proceedings. By analyzing microbial diversity in water samples and drowned animal lung tissues, the study sheds light on how environmental factors and victim-related variables influence microbial communities. The findings not only advance our understanding of forensic microbiology but also offer practical implications for improving investigative techniques in cases of drowning.

Gill Health in Fish Farmed in Recirculating Aquaculture Systems (RAS): A Review.

Recirculating Aquaculture Systems (RAS) have been proposed as the future of aquaculture, because they can be used anywhere regardless of access to water, they offer high level of control over farming environment, including biosecurity, and are considered to be sustainable. However, despite of continuous development, there can be still issues with water quality affecting gill health of fish farmed in these systems. This review provides an overview of fish gill structure and gill immune response, and discusses the known impacts of RAS on gill health. Several experimental studies have inadequately reported conditions, particularly water quality, making it difficult to determine if the observed effects were due to water quality issues or RAS system itself. It is crucial for studies investigating the impact of RAS on fish to report water quality during the study. Furthermore, assessments of RAS effects on gill health should include sufficient independent replicates and flow through controls using a common water source. Various methods have been used to assess gill health in RAS, including gill histology, presence of pathogens, gene expression in the gills and gill microbiome analysis. Differences in gill health in fish from RAS and a flow through system have been shown for a number of freshwater and marine fish species. However, these results have been inconsistent across studies, and some results have been challenging to interpret as indicators of gill health. Holistic studies including a number of different methods to assess fish gills would give more conclusive results. More research is needed, in particular, on brackish and marine RAS, to fully understand their impacts on gill health.

Oral Microbiome Research in Biopsy Samples of Oral Potentially Malignant Disorders and Oral Squamous Cell Carcinoma and Its Challenges

This study aims to evaluate the potential benefits and challenges of integrating oral microbiome research into the clinical management of oral potentially malignant disorders (OPMD) and oral squamous cell carcinoma (OSCC). The oral microbiome has gained significant attention for its role in the pathogenesis and progression of these conditions, with emerging evidence suggesting its value as a diagnostic and prognostic tool. By critically analyzing current evidence and methodological considerations, this manuscript examines whether microbiome analysis in biopsy samples can aid in the early detection, prognosis, and management of OPMD and OSCC. The complexity and dynamic nature of the oral microbiome require a multifaceted approach to fully understand its clinical utility. Based on this review, we conclude that studying the oral microbiome in this context holds significant promise but also faces notable challenges, including methodological variability and the need for standardization. Ultimately, this manuscript addresses the question, “Should such research be undertaken, given the intricate interactions of various factors and the inherent obstacles involved?”, and also emphasizes the importance of further research to optimize clinical applications and improve patient outcomes.

Omnigene-Guttm ensures fecal microbiome stability in the pediatric population

Increasing evidence exists that the gut microbiome influences toxicity as well as outcomes in a variety of cancers. To investigate the role of the gut microbiome in pediatric neuro-oncology, microbiome analysis has been included in multiple prospective pediatric neuro-oncology clinical trials (NCT05009992, NCT04732065, NCT04775485). In these trials, the OMNIgene-GUTtm preservation tubes are used for the collection of the feces. OMNIgene-GUTtm has demonstrated reliability in preserving the composition of the gut microbiome in adults; however, its validation for use in the pediatric population remains limited. Therefore, we compared the quality of the DNA by 16S rRNA gene sequencing after various methods of stabilizing fecal samples in pediatric populations, from the direct freeze method at − 80 °C to preserving samples with OMNIgene-GUTtm at room temperature for various durations. Our results showed that there were no statistically significant differences between the alpha-diversity, and beta-diversity. However, pairwise differential abundance analyses demonstrated that OMNIgene-GUT™ is superior in maintaining microbial community structure compared to storing samples without any preservation method. With the OMNIgene-GUTtm’s stabilization of the fecal samples being superior and its ease-of-use benefits, it proves to be a valid and ideal method of stabilizing fecal samples for current and future pediatric clinical trials.

Sexually dimorphic metabolic effects of a high fat diet on knee osteoarthritis in mice

BackgroundWomen have a higher risk of developing osteoarthritis (OA) than men, including with obesity. To better understand this disparity, we investigated sex differences in metabolic and inflammatory factors associated with OA using a diet-induced mouse model of obesity. We hypothesized that 20 weeks of high-fat diet (HFD) would induce sexually dimorphic changes in both systemic and local risk factors of knee OA.MethodsMale and female C57BL/6J mice were fed Chow or HFD from 6 to 26 weeks of age (n = 12 per diet and sex). We performed broad metabolic phenotyping, 16 S gut microbiome analysis, targeted gene expression analysis of synovium-infrapatellar fat tissue, targeted gene expression and proteomic analysis of articular cartilage, chondrocyte metabolic profiling, and OA histopathology. Two-way ANOVA statistics were utilized to determine the contribution of sex and diet and their interaction on outcomes.ResultsMice fed HFD weighed 1.76-fold (p < 0.0001) and 1.60-fold (p < 0.0001) more than male and female Chow cohorts, respectively, with both sexes reaching similar body fat levels (male: 43.9 ± 2.2%; female: 44.1 ± 3.8%). HFD caused greater cartilage pathology (p < 0.024) and synovial hyperplasia (p < 0.038) versus Chow in both sexes. Cartilage pathology was greater in male versus female mice (p = 0.048), and only male mice developed osteophytes with HFD (p = 0.044). Both sexes exhibited metabolic inflexibility on HFD, but only male mice developed glucose intolerance (p < 0.0001), fatty liver (p < 0.0001), and elevated serum amylase (p < 0.0001) with HFD versus Chow. HFD treatment caused sex-dependent differences in gut microbiota beta diversity (p = 0.01) and alteration in specific microbiome clades, such as a HFD-dependent reduction in abundance of Bifidobacterium only in male mice. In knee synovium and infrapatellar fat tissue, HFD upregulated the expression of pro-inflammatory and pro-fibrotic genes predominantly in female mice. In cartilage, lipid metabolism proteins were more abundant with HFD in male mice, whereas proteins involved in glycolysis/gluconeogenesis and biosynthesis of amino acids were greater in cartilage of female mice. Sex-dependent metabolic differences were observed in cartilage from young, healthy mice prior to pubertal maturation, but not in primary juvenile chondrocytes studied in vitro.ConclusionsHFD induced numerous sex differences in metabolic and inflammatory outcomes, especially in joint tissues, suggesting that sex-specific cellular processes are involved during development of early-stage OA with obesity.

Wild-Mouse-Derived Gut Microbiome Transplantation in Laboratory Mice Partly Alleviates House-Dust-Mite-Induced Allergic Airway Inflammation

Laboratory mice are instrumental for preclinical research but there are serious concerns that the use of a clean standardized environment for specific-pathogen-free (SPF) mice results in poor bench-to-bedside translation due to their immature immune system. The aim of the present study was to test the importance of the gut microbiota in wild vs. SPF mice for evaluating host immune responses in a house-dust-mite-induced allergic airway inflammation model without the influence of pathogens. The wild mouse microbiome reduced histopathological changes and TNF-α in the lungs and serum when transplanted to microbiota-depleted mice compared to mice transplanted with the microbiome from SPF mice. Moreover, the colonic gene expression of Gata3 was significantly lower in the wild microbiome-associated mice, whereas Muc1 was more highly expressed in both the ileum and colon. Intestinal microbiome and metabolomic analyses revealed distinct profiles associated with the wild-derived microbiome. The wild-mouse microbiome thus partly reduced sensitivity to house-dust-mite-induced allergic airway inflammation compared to the SPF mouse microbiome, and preclinical studies using this model should consider using both ‘dirty’ rewilded and SPF mice for testing new therapeutic compounds due to the significant effects of their respective microbiomes and derived metabolites on host immune responses.

Therapeutic potential of Xihuang Pill in colorectal cancer: Metabolomic and microbiome-driven approaches.

The Xihuang Pill (XHP), a venerated traditional Chinese medicine, has demonstrated significant anti-cancer capabilities. Despite its proven efficacy, the scarcity of comprehensive pharmacological studies limits the widespread application of XHP. This research endeavor seeks to demystify the therapeutic underpinnings of XHP, particularly in the realm of colorectal cancer (CRC) therapy. In this study, mice harboring CT26 tumors were divided into four groups, each administered with either XHP monotherapy, 5-fluorouracil (5-FU), or a combination of both. The tumor growth trajectory was closely monitored to evaluate the effectiveness of these anti-neoplastic interventions. Advanced techniques, including 16S-rDNA gene sequencing and ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), were harnessed to scrutinize the gut microbiota and serum metabolite profiles. Immunohistochemical assays were employed to gauge the expression levels of CD4, CD8, and Foxp3, thereby providing insights into the dynamics of tumor-infiltrating lymphocytes within the tumor microenvironment. Our findings indicate that XHP effectively suppresses the initiation and progression of colorectal tumors. The combinatorial therapy of XHP with 5-FU exhibited an enhanced inhibitory effect on tumor growth. Metabolic profiling revealed that XHP induced notable metabolic shifts, particularly impacting pathways such as steroid hormone synthesis, arachidonic acid metabolism, purine biosynthesis, and renin secretion. Notably, 17α-ethinyl estradiol and α-ergocryptine were identified as serum metabolites with the most substantial increase following XHP administration. Analysis of the gut microbiome suggested that XHP promoted the expansion of specific bacterial taxa, including Lachnospiraceae_NK4A136_group, Clostridiales, Desulfovibrionaceae, and Anaerotignum_sp., while suppressing the proliferation of others such as Ligilactobacilus, Lactobacillus_taiwanensis, and Candidatus_saccharimonas. Immunohistochemical staining indicated an upregulation of CD4 and CD8 post-XHP treatment. This study delineates a potential mechanism by which XHP inhibits CRC tumorigenesis through modulating the gut microbiota, serum metabolites, and reshaping the tumor immune microenvironment in a murine CRC model. These findings contribute to a more profound understanding and potentially broaden the clinical utility of XHP in oncology.

Involvement of Mediterranean fever gene mutations in colchicine-responsive enterocolitis: a retrospective cohort study

The involvement of Mediterranean fever (MEFV) gene mutations in patients with inflammatory bowel disease unclassified (IBDU) remains unclear. This study aimed to determine the clinical characteristics and responsiveness to colchicine in Japanese patients with IBDU carrying MEFV mutations. In this retrospective cohort study, we examined MEFV mutations using gene analysis, clinical information, and colchicine responsiveness. Furthermore, we examined cytokine production in exon 2-mutated THP-1cells (a monocytic cell line) and microbiome analysis. Of the 396 patients diagnosed with IBDU, 60.1% had MEFV mutations. Exon 2 mutations were the most common (83.7%). Among patients with available clinical information, 43.3% of patients with IBDU had typical Familial Mediterranean fever (FMF). The efficacy of colchicine in patients with IBDU carrying MEFV mutations was 84.6%. Significant differences were noted in the production of inflammatory; cytokines between THP-1cells with and without MEFV mutations. Microbial compositions differed between patients with IBDU carrying MEFV mutations and patients with IBD and healthy controls. Patients with IBDU carrying MEFV mutations responded well to colchicine treatment. A notable subset of patients met the criteria for typical FMF. Alterations in intestinal microbiota may contribute to disease pathogenesis. This work was supported by the Japan Agency for Medical Research and Development (21ek0410057h0003), a grant from the Uehara Memorial Foundation, and the Health and Labour Sciences Research Grants for research on intractable diseases from the Ministry of Health, Labour and Welfare (MHLW) of Japan (Investigation and Research for Intractable Inflammatory Bowel Disease; Grant Number 20316729).

The characteristics of pharyngea microbiological in children with obstructive sleep apnea

Objective:This study aimed to explore the possible pathogenesis of OSA from the perspective of microbiology by evaluate the change in pharyngeal microbiome of OSA children, and provide new ideas for clinical prevention, diagnosis and treatment. Methods:Randomly enrolled 20 children with OSA as OSA group and 20 children without OSA as control group. The swallow swab of each children been collected. Using 16srDNA sequencing to investigate the characteristics of pharyngeal microbiome. Results:The α-diversity showed that the Chao1and Observe-Otus index has significantly increased in the OSA group, and the β-diversity was significantly different between the two groups. The relative abundance of Haemophilus（Proteobacteria） increased but that of Veillonella（member of Firmicutes） and Prevotella-7 and Prevotella（member of Bacteroidota） decreased in the OSA group compared to control group. Conclusion:The pharyngeal microbial richness are decreased significantly and composition are disrupted in children with OSA. This microbiome analysis provides a new understanding about the pathogenesis of OSA in children.