16S rRNA Amplicon Research Articles

Faecal microbiota and cytokine profiles of rural Cambodian infants linked to diet and diarrhoeal episodes

The gut microbiota of infants in low- to middle-income countries is underrepresented in microbiome research. This study explored the faecal microbiota composition and faecal cytokine profiles in a cohort of infants in a rural province of Cambodia and investigated the impact of sample storage conditions and infant environment on microbiota composition. Faecal samples collected at three time points from 32 infants were analysed for microbiota composition using 16S rRNA amplicon sequencing and concentrations of faecal cytokines. Faecal bacterial isolates were subjected to whole genome sequencing and genomic analysis. We compared the effects of two sample collection methods due to the challenges of faecal sample collection in a rural location. Storage of faecal samples in a DNA preservation solution preserved Bacteroides abundance. Microbiota analysis of preserved samples showed that Bifidobacterium was the most abundant genus with Bifidobacterium longum the most abundant species, with higher abundance in breast-fed infants. Most infants had detectable pathogenic taxa, with Shigella and Klebsiella more abundant in infants with recent diarrhoeal illness. Neither antibiotics nor infant growth were associated with gut microbiota composition. Genomic analysis of isolates showed gene clusters encoding the ability to digest human milk oligosaccharides in B. longum and B. breve isolates. Antibiotic-resistant genes were present in both potentially pathogenic species and in Bifidobacterium. Faecal concentrations of Interlukin-1alpha and vascular endothelial growth factor were higher in breast-fed infants. This study provides insights into an underrepresented population of rural Cambodian infants, showing pathogen exposure and breastfeeding impact gut microbiota composition and faecal immune profiles.

PSX-14 Effect of starch supplementation on rumen bacterial abundance in Bos taurus taurus steers consuming low-quality forage

Abstract Low-quality forage provides a limited supply of ruminally available nitrogen and is generally resistant to digestion, inhibiting microbial growth in the rumen and ultimately decreasing forage utilization. Supplementation with protein is well documented to increase forage utilization by promoting microbial growth. Even when protein requirements are met, additional energy, often provided as starch, may be required to meet performance expectations. The objective of this study was to evaluate the effect of increasing starch supplementation on rumen bacterial populations in Bos taurus taurus steers supplemented protein and consuming low-quality forage. Ruminal samples were collected from ruminally-cannulated Angus steers [n = 5; body weight (BW) = 375 ± 45 kg] used in a 4 × 4 Latin square fed a basal diet of King Ranch Bluestem Hay (3.5% CP, 73% NDF) ad libitum, and supplemented with one of four isonitrogenous (130 mg N/kg BW) supplements with increasing starch (2% starch = 100% soybean meal; 20% starch = 73% soybean meal, 26.2% corn, 0.8% urea; 38% starch = 47% soybean meal, 51.6% corn, 1.4% urea; 56% starch = 19% soybean meal, 78.6% corn, 2.4% urea). Rumen content samples were collected via rumen cannula 4 h after supplementation and immediately frozen. Microbial DNA was extracted; full length 16S rRNA amplicon libraries were prepared using Kinnex kits and sequenced on the PacBio Revio platform. Reads were analyzed using DADA2 and QIIME2. Statistical analysis was performed with the GLIMMIX procedure of SAS 9.4 (SAS Inst. Inc., Cary, NC). Firmicutes and Bacteriodata were the most abundant phyla in all treatments (82% and 11%, respectively), and were not affected by starch supplementation (P ≥ 0.45). Actinobacteriota and Euryarchaeota abundance responded quadratically (P < 0.01) to increasing starch provision. Christensenellaceae R-7 group, Rikenellaceae RC9 gut group, Lachnospiraceae XPB1014 group, Butyrvibrio, Lachnospiraceae NK3A20 group, and Prevotella represented the most abundant genera for all treatments. Increasing supplemental starch resulted in a quadratic response (P < 0.01) for abundance of Christensenellaceae R-7 and Lachnospiraceae XPB1014 groups. Relative abundance of Butyrvibrio was 4.05% for steers fed 2% starch supplement and increased to 18.24 and 11.85% for 20 and 38% starch, respectively and then decreased to 4.34% for steers fed 56% starch supplement (quadratic, P < 0.01) Abundance of Pseudobutyrvibrio followed a similar but less dramatic pattern (quadratic, P < 0.01). In contrast, abundance of Streptococcus and Methanobrevibacter decreased with the first two levels of starch and then increased with 56% (quadratic; P < 0.01). In conclusion, responses to the first two increments of starch supplementation suggest increased fibrolytic and decreased methanogenic activity. These results are supported by the previously reported increase in TDOMI for 20 and 38% starch 49.0 and 48.2 g/kg MBW, respectively versus 45.7 and 44.3 g/kg MBW for 2 and 56%, respectively.

PSX-15 Effect of starch supplementation on rumen bacterial abundance in Bos taurus indicus steers consuming low-quality forage

Abstract Deficient ruminally available nitrogen caused by a basal diet of low-quality forage can limit microbial growth in the rumen and decrease forage utilization. Protein supplementation is well-documented to improve forage digestibility and performance. Additionally, starch has been evaluated to increase energy supply to grazing cattle when the forage cannot support adequate levels of production. However, most research in this area has been conducted in Bos taurus taurus, especially the response of the rumen microbiome to supplementation. Accordingly, the objective of this study was to evaluate the effects of increasing starch supplementation on rumen bacterial populations in Bos taurus indicus steers supplemented protein and consuming low-quality forage. Ruminal samples were collected from five ruminally-cannulated Brahman steers (BW 420 ± 55 kg) used in a 4×4 Latin square fed a basal diet of King Ranch Bluestem Hay (3.5% CP and 73% NDF) provided ad libitum, and one of four isonitrogenous (130 mg N/kg BW) supplements with increasing levels of starch (2% starch= 100% soybean meal; 20% starch= 73% soybean meal, 26.2% corn, 0.8% urea; 38% starch = 47% soybean meal, 51.6% corn, 1.4% urea; 56% starch= 19% soybean meal, 78.6% corn, 2.4% urea). Rumen content samples were collected via rumen cannula at 4h post-feeding. Microbial DNA was extracted; full length 16S rRNA amplicon libraries were prepared using Kinnex kits and then sequenced on the PacBio Revio platform. Reads were analyzed using DADA2 and QIIME2. Statistical analysis was performed with the GLIMMIX procedure of SAS 9.4 (SAS Inst. Inc., Cary, NC). Firmicutes, Bacteroidata, Actinobacteriota, Verrucomicrobiota, and Euryarchaeota were the most abundant phyla in all treatments (83, 10, 3, 0.4, and 1%, respectively), and were not affected by treatment (P ≥ 0.10). Christensenellaceae R-7 group, Rikenellaceae RC9 gut group, Lachnospiraceae-XPB1014-group, Butyrvibrio, Lachnospiraceae-NK3A20-group, and Prevotella represented the most abundant genera for all treatments (7, 5, 11, 9, 7, and 3%, respectively). Lachnospiraceae-XPB1014-group and Lachnospiraceae-NK3A20-group responded quadratically (P ≤ 0.02) to increasing starch with decreased abundance for the 20% and 38% starch supplement. Comparatively, relative abundance of Butyrvibrio responded quadratically (P < 0.01) with increased abundance in the 20% and 38% starch supplement. Pseudobutyrvibrio increased with supplementation of 20% starch and then decreased as starch supplementation increased (quadratic, P = 0.04). In conclusion, supplementation of starch elicited changes in microbial relative abundance at the genus level with the response in Butyrivibrio suggesting greater fibrolytic activity. Forage and total digestible NDF intake in these steers decreased with increased starch supplementation; however, TDOMI was not affected by starch provision.

Microbial consortia in commercial salmon products: A comparative study using cultivation-dependent and high-throughput sequencing approaches

Despite the widespread consumption of salmon products, an all-round microbiological survey on fresh salmon and smoked salmon products sold on the market directly to consumers remains to be undertaken. The present study conducted a comprehensive microbial investigation employing both conventional cultivation-based techniques and 16S rRNA amplicon sequencing. A total of 120 fresh salmon and 95 smoked salmon products were systematically collected across all four seasons (spring, summer, fall, and winter), accompanied by temperature assessments of each product’s interior. The average aerobic plate counts for fresh salmon and smoked salmon were 3.63 log and 1.36 log CFU/g, respectively, and the highest seasonal concentrations occurred in summer. The average coliform count was higher in fresh salmon (1.50 log CFU/g) than in smoked salmon products (1.12 log CFU/g). Although smoked salmon exhibited CFU counts similar to those of fresh salmon according to culture-based methods, 16S sequencing unveiled a distinct microbial composition. Photobacterium and Pseudomonas, recognized as major spoilage-related microorganisms, were the predominant genera in all products, but especially in fresh salmon. Smoked salmon exhibited diverse variations in bacterial composition compared to fresh salmon. Three types of lactic acid bacteria—Leuconostoc, Lactobacillus, and Lactococcus—were found to be significantly more abundant in smoked salmon than in fresh salmon. The outcomes of this study contribute to an enhanced understanding of retail salmon products, shedding light on crucial areas for investigation and suggesting potential interventions regarding both sales and production of salmon products.

Interaction and effects of temperature preference under a controlled environment on the diversity and abundance of the microbiome in Lutzomyia longipalpis (Diptera: Psychodidae)

Characterization of the temperature effects on the abundance and richness of the microbiota of Lutzomyia longipalpis, insect vector of Leishmania infantum in America, is an aspect of pivotal importance to understand the interactions between temperature, bacteria, and Leishmania infection. We developed and used a customized device with a temperature gradient (21–34 °C) to assess the temperature preferences of wild females of Lu. longipalpis collected in a rural area (Ricaurte, Cundinamarca, Colombia). Each replicate consisted of 50 females exposed to the gradient for an hour. At the end of the exposure time, insects were collected and separated by the temperature ranges selected varying from 21 °C to 34 °C. They were organized in 17 pools from which total DNA extracts were obtained, and samples were subjected to 16S rRNA amplicon sequencing analyzes. The most abundant phyla across the different temperature ranges were Proteobacteria (17.22–90.73 %), Firmicutes (5.99–77.21 %) and Actinobacteria (1.56–59.85 %). Results also showed an abundance (30 % to 57.36 %) of Pseudomonas (mainly at temperatures of 21–29 °C and 34 °C) that decreased to 6.55 %-13.20 % at temperatures of 31–33 °C, while Bacillus increase its abundance to 67.24 % at 29–33 °C. Serratia also had a greater representation (49.79 %), specifically in sand flies recovered at 25–27 °C. No significant differences were found at α-diversity level when comparing richness using the Shannon-Wiener, Simpson, and Chao1 indices, while β-diversity differences were found using the Bray-Curtis index (F-value of 3.5073, p-value < 0.013, R-squared of 0,4889), especially in the groups of Lu. longipalpis associated at higher temperatures (29–33 °C). It was also possible to detect the presence of endosymbionts such as Spiroplasma and Arsenophonus in the range of 29–33 °C. Rickettsia was only detected in Lu. longipalpis sand flies recovered between 25–27 °C. It was possible to characterize Lu. longipalpis microbiota in response to intraspecific temperature preferences and observe changes in bacterial communities and endosymbionts at different ranges of said environmental variable, which may be important in its vector competence and environmental plasticity to adapt to new climate change scenarios.

Adaptation towards catabolic biodegradation of trace organic contaminants in activated sludge

Trace organic contaminants (TrOCs) are omnipresent in wastewater treatment plants (WWTPs), yet, their removal during wastewater treatment is oftentimes incomplete and underlying biotransformation mechanisms are not fully understood. In this study, we elucidate how different factors, including pre-exposure levels and duration, influence microbial adaptation towards catabolic TrOC biodegradation and its potential role in biological wastewater treatment. Four sequencing batch reactors (SBRs) were operated in parallel in three succeeding phases, adding and removing a selection of 26 TrOCs at different concentration levels. After each phase of SBR operation, a series of batch experiments was conducted to monitor biotransformation kinetics of those same TrOCs across various spike concentrations. For half of our test TrOCs, we detected increased biotransformation in sludge pre-exposed to TrOC concentrations ≥5 µg L−1 over a 30-day period, with most significant differences observed for the insect repellent DEET and the artificial sweetener saccharin. Accordingly, 16S rRNA amplicon sequencing revealed enrichment of taxa that have previously been linked to catabolic biodegradation of several test TrOCs, e.g., Bosea sp. and Shinella sp. for acesulfame degradation, and Pseudomonas sp. for caffeine, cyclamate, DEET, metformin, paracetamol, and isoproturon degradation. We further conducted shotgun metagenomics to query for gene products previously reported to be involved in the TrOCs’ biodegradation pathways. In the future, directed microbial adaptation may be a solution to improve bioremediation of TrOCs in contaminated environments or in WWTPs.

Sward type alters enteric methane emissions, nitrogen output and the relative abundance of the rumen microbial ecosystem in sheep.

Observed improvements in animal and sward performance, coupled with a desire for more sustainable pasture-based feeding systems, has triggered a surge in the implementation of more botanically diverse pastures. However, thus far, there has been limited research investigating the effects of botanically diverse sward types on enteric methane (CH4) or nitrogen (N) excretion, alongside the ruminal microbiota and fermentation profile, in sheep. Hence, this study investigates the effect of sward type on CH4 production and N excretion, in addition to assessing the rumen microbiome, volatile fatty acid proportions, and ammonia nitrogen (NH3-N) concentration in sheep. A 5 × 5 Latin square design experiment was implemented to investigate 5 dietary treatments; perennial ryegrass (Lolium perenne L.; PRG) only or PRG plus white clover (Trifolium repens L.; PRG + WC), red clover (Trifolium pratense L.; PRG + RC), chicory (Chicorium intybus L.; PRG + Chic) or plantain (Plantago lanceolata L.; PRG + Plan). Diets were mixed at a ratio of 75% PRG and 25% of the respective companion forage and 100% PRG for the PRG treatment, on a dry matter basis. Twenty castrated male sheep were housed in metabolism crates across 5 feeding periods. Methane measurements were acquired utilizing portable accumulation chambers. Rumen fluid was harvested using a transoesophageal sampling device. Microbial rumen DNA was extracted and subjected to 16S rRNA amplicon sequencing and fermentation analysis. Data were analyzed using PROC MIXED in SAS. Results show that animals consuming PRG + WC ranked lower for CH4 production (g/d) than sheep offered PRG, PRG + Chic or PRG + Plan (P < 0.01) while the addition of any companion forage ranked CH4 yield (g/kg dry matter intake (DMI)) lower (P < 0.001) than PRG. There was a moderate positive correlation between DMI and CH4 (g/d; r = 0.51). Ruminal NH3-N was lowest in animals consuming the PRG diet (P < 0.01). There was a greater abundance of Methanobrevibacter and reduced abundance of Methanosphaera (P < 0.001) in sheep offered PRG, compared with any binary sward. On average, herb diets (PRG + Chic or PRG + Plan) reduced the urinary nitrogenconcentration of sheep by 34% in comparison to legume diets (PRG + WC or PRG + RC) and 13% relative to the PRG diet (P < 0.001). Sheep offered PRG + Chic had a greater dietary nitrogen use efficiency than PRG + RC (P < 0.05). This study demonstrates the potential for sward type to influence rumen function and the microbial community, along with CH4 and N output from sheep.

Multi-omics study unravels gut microbiota and metabolites alteration in patients with Wilson's disease

Hepatolenticular degeneration (HLD), also known as Wilson's disease (WD), is a rare autosomal recessive disorder regarding copper metabolism. Whether gut microbiota imbalance is involved in developing HLD remains unknown. A comprehensive 16S rRNA amplicon sequencing, metagenomic sequencing, and metabonomic analysis were undertaken in patients with WD to analyze the composition and function profiles of gut microbiota in patients with WD. The data demonstrated differences in gut microbiota and metabolic pathways between WD patients and normal individuals, significantly decreasing bacterial richness and diversity. The levels of Selenomonaceae and Megamonas in WD patients are significantly higher than those in healthy individuals. The relative abundances of Roseburia inulinivorans in patients with WD are lower than in healthy individuals. Compared with healthy people, the level of metabolites in patients with WD is abnormal. Leucylproline, 5-Phenylvaleric Acid and N-Desmethylclobazam, which have nutritional and protective effects, are significantly reduced fecal metabolites in patients with WD. D-Gluconic acid, which can chelate metal ions, may be a potential treatment for WD. The positive correlation it demonstrates with Alistipes indistinctus and Prevotella stercora indicates potential bacteria able to treat WD. These metabolites are mainly related to the biosynthesis of antibiotics, alpha-linolenic acid metabolism, one carbon pool by folate, nicotinate and nicotinamide metabolism. In conclusion, the data from this study elucidate novel mechanisms describing how abnormal gut miccrobiota contribute to the pathogenesis of WD and outlines new molecules for the treatment of WD.

Co-treatment of agri-food waste streams using black soldier fly larvae (Hermetia illucens L.): A sustainable solution for rural waste management

The management of rural waste, particularly agri-food waste, poses a major challenge to the ecosystem health. This study investigated the efficacy of black soldier fly larvae (Hermetia illucens L., BSFL) bioconversion for agri-food waste under independent treatment or co-treatment strategies using chicken manure and food waste as a model system. The results showed a synergistic effect of co-treating agri-food waste from different sources. The co-treatment strategy enhanced bioconversion efficiency, resulting in a 1.31-fold waste reduction rate and a 1.93-fold bioconversion rate. Additionally, larval growth performance and biomass quality of BSFL were improved, while lauric acid and oleic acid were enriched in the larval fat from the co-treatment strategy. 16S rRNA amplicon sequencing revealed that the co-treatment strategy reshaped both the residue and larval gut microbiota, with distinct enrichment of taxonomical biomarkers. Furthermore, under this strategy, metabolic functions of the residue microbiota were significantly activated, especially carbohydrate, amino acid, and lipid metabolism were enhanced by 16.3%, 23.5%, and 20.2%, respectively. The early colonization of lactic acid bacteria (Weisella and Aerococcus) in the residue, coupled with a symbiotic relationship between Enterococcus in the larval gut and the host, likely promoted organic matter degradation and larval growth performance. Scaling up the findings to a national level in China suggests that the co-treatment strategy can increase waste reduction quantity by 86,329 tonnes annually and produce more larval protein and fat with a market value of approximately US$237 million. Therefore, co-treatment of agri-food waste streams using BSFL presents a sustainable solution for rural waste management that potentially contributes to the achievement of SDG2 (Zero Hunger), SDG3 (Good Health and Well-Being), and SDG12 (Responsible Consumption and Production).

Treatment of Anaerobic Digester Liquids via Membrane Biofilm Reactors: Simultaneous Aerobic Methanotrophy and Nitrogen Removal.

Anaerobic digestion (AD) produces useful biogas and waste streams with high levels of dissolved methane (CH4) and ammonium (NH4+), among other nutrients. Membrane biofilm reactors (MBfRs), which support dissolved methane oxidation in the same reactor as simultaneous nitrification and denitrification (ME-SND), are a potential bubble-less treatment method. Here, we demonstrate ME-SND taking place in single-stage, AD digestate liquid-fed MBfRs, where oxygen (O2) and supplemental CH4 were delivered via pressurized membranes. The effects of two O2 pressures, leading to different O2 fluxes, on CH4 and N removal were examined. MBfRs achieved up to 98% and 67% CH4 and N removal efficiencies, respectively. The maximum N removal rates ranged from 57 to 94 mg N L-1 d-1, with higher overall rates observed in reactors with lower O2 pressures. The higher-O2-flux condition showed NO2- as a partial nitrification endpoint, with a lower total N removal rate due to low N2 gas production compared to lower-O2-pressure reactors, which favored complete nitrification and denitrification. Membrane biofilm 16S rRNA amplicon sequencing showed an abundance of aerobic methanotrophs (especially Methylobacter, Methylomonas, and Methylotenera) and enrichment of nitrifiers (especially Nitrosomonas and Nitrospira) and anammox bacteria (especially Ca. Annamoxoglobus and Ca. Brocadia) in high-O2 and low-O2 reactors, respectively. Supplementation of the influent with nitrite supported evidence that anammox bacteria in the low-O2 condition were nitrite-limited. This work highlights coupling of aerobic methanotrophy and nitrogen removal in AD digestate-fed reactors, demonstrating the potential application of ME-SND in MBfRs for the treatment of AD's residual liquids and wastewater. Sensor-based tuning of membrane O2 pressure holds promise for the optimization of bubble-less treatment of excess CH4 and NH4+ in wastewater.

Metagenomic analysis unveiled the response of microbial community and antimicrobial resistome in natural water body to duck farm sewage

Sewages from duck farms are often recognized as a major source of antimicrobial resistance and pathogenic bacteria discharged to natural water bodies, but few studies depicted the dynamic changes in resistome and microbial communities in the rivers under immense exposure of sewage discharge. In this study, we investigated the ecological and environmental risks of duck sewages to the rivers that geographically near to the duck farms with short-distance (<1 km) using 16S rRNA amplicon and metagenomic sequencing. The results showed that a total of 20 ARG types were identified with abundances ranged from 0.61 to 1.33 cpc. Of note, the genes modulate resistances against aminoglycoside, bacitracin and beta-lactam were the most abundant ARGs. Limnohabitans, Fluviibacter and Cyanobium were the top 3 predominant genera in the microbial community. The alpha diversity of overall microbial community decrease while the abundance of pathogen increase during the input of sewage within 200 m. Sul1 and bacA were the dominant ARGs brought from duck farm sewage. The community variations of ARGs and microbiome were primarily driven by pH and temperature. Total phosphorus was significantly correlated to alpha diversity and top 30 ARGs subtype. Stochastic processes was the dominated microbial assembly pattern and did not be altered by sewage. We also highlighted the ecological risk caused by blaGES which possibly could be mitigated by Cyanobacteria, and the natural water body can purify partial ARGs as well as microbiome from duck farms sewage. These findings expanded our knowledge regarding the ecological risks by wastes from the livestock farm, and underscoring the necessity to monitor ARGs in farm-surrounding water bodies.

Adaptation of a microbial consortium to pelagic Sargassum modifies its taxonomic and functional profile that improves biomethane potential.

In recent years, pelagic Sargassum has invaded the Caribbean coasts, and anaerobic digestion has been proposed as a sustainable management option. However, the complex composition of these macroalgae acts as a barrier to microbial degradation, thereby limiting methane production. Microbial adaptation is a promising strategy to improve substrate utilization and stress tolerance. This study aimed to investigate the adaptation of a microbial consortium to enhance methane production from the pelagic Sargassum. Microbial adaptation was performed in a fed-batch mode for 100days by progressive feeding of Sargassum. The evolution of the microbial community was analyzed by high-throughput sequencing of 16S rRNA amplicons. Additionally, 16S rRNA data were used to predict functional profiles using the iVikodak platform. The results showed that, after adaptation, the consortium was dominated by the bacterial phyla Bacteroidota, Firmicutes, and Atribacterota, as well as methanogens of the families Methanotrichaceae and Methanoregulaceae. The abundance of predicted genes related to different metabolic functions was affected during the adaptation stage when Sargassum concentration was increased. At the end of the adaptation stage, the abundance of the predicted genes increased again. The adapted microbial consortium demonstrated a 60% increase in both biomethane potential and biodegradability index. This work offers valuable insights into the development of treatment technologies and the effective management of pelagic Sargassum in coastal regions, emphasizing the importance of microbial adaptation in this context.

The Microleaks study: 16S community profile and metagenomic shotgun sequencing signatures associated with anastomotic leak

Anastomotic leaks (AL) are the most severe complications of colorectal surgery. The cause of AL is unclear, but recent studies have implicated the intestinal microbiota in its development. We aimed to determine whether there is an identifiable microbial pattern in the mucosal microbiota associated with AL. A pragmatic series of 162 patients undergoing colorectal resection with anastomosis had swabs taken from the proximal and distal mucosa of the bowel resection immediately after the tissue was excised. DNA was extracted for 16S rRNA amplicon gene sequencing and a subset for metagenomic shotgun sequencing (MGS). The AL rate in the cohort was approximately 15% (25/162). The alpha diversity measures from the intraoperative swabs were all significantly increased for AL, and there were significant differences in the beta diversity measures for AL from both the 16S and MGS datasets. The predictive power of AL was more sensitive when both proximal and distal communities were considered, and the species-level classifier AUC-ROC was stronger for the MGS dataset than for the 16S data (AUC = 0.92 and 0.76, respectively). We also report, for the first time, the functional changes in intraoperative AL microbes and noted an increase in the relative abundance of pathways with fermentation end products. This result was also found in our murine model of anastomoses (n = 20). At the time of surgery, the mucosal microbiota of the anastomotic extremities exhibits subtle differences at the species level and altered fermentation capacity, which may be associated with AL outcomes. A greater understanding of these insights could improve AL prognosis and preoperative management to reduce the occurrence of this life-threatening condition.

Metabarcoding study of potential pathogens and zoonotic risks associated with dog feces in Seoul, South Korea.

A significant portion of South Korea's population, approximately a quarter, owns pets, with dogs being the most popular choice among them. However, studies analyzing the fecal organism communities of dogs in South Korea are lacking, and limited efforts have been exerted to identify pathogens with potential zoonotic implications. Therefore, this study aimed to investigate potential pathogens using metabarcoding analysis and evaluate the risk of zoonotic diseases in dog feces in Seoul, South Korea. Fecal samples were collected from both pet and stray dogs in the Mapo district of Seoul. Next-generation sequencing (NGS) was utilized, employing 16S rRNA amplicon sequencing to identify prokaryotic pathogens, and 18S rRNA amplicon sequencing for eukaryotic pathogens. The data obtained from the QIIME2 pipeline were subjected to various statistical analyses to identify different putative pathogens and their compositions. Significant variations in microbiota composition were found between stray and pet dogs, and putative prokaryotic and eukaryotic pathogens were identified. The most prevalent putative bacterial pathogens were Fusobacterium, Helicobacter, and Campylobacter. The most prevalent putative eukaryotic pathogens were Giardia, Pentatrichomonas, and Cystoisospora. Interestingly, Campylobacter, Giardia, and Pentatrichomonas were found to be significantly more prevalent in stray dogs than in pet dogs. The variation in the prevalence of potential pathogens in dog feces could be attributed to environmental factors, including dietary variances and interactions with wildlife, particularly in stray dogs. These factors likely contributed to the observed differences in pathogen occurrence between stray and pet dogs. This study offers valuable insights into the zoonotic risks associated with dog populations residing in diverse environments. By identifying and characterizing putative pathogens in dog feces, this research provides essential information on the impact of habitat on dog-associated pathogens, highlighting the importance of public health planning and zoonotic risk management.

Microbial Community Structure and Metabolism of Xinjiang Fine-Wool Sheep based on High-Throughput Sequencing Technology.

It turns out that the more than trillion microorganisms living in the host's digestive tract are crucial for maintaining nutrient intake, environmental suitability, and physiological mechanism. Xinjiang fine-wool sheep is an exclusive breed for wool in China, which has excellent stress tolerance. In this study, we collected feces and blood samples of 20 Xinjiang fine-wool sheep under the same genetic characteristics, the Fine-Wool Sheep (FWS) group and the Control Fine-Wool Sheep (CFWS) group were set up according to the differs in phenotypic characteristics of their wool. By 16S rRNA amplicon sequence, ITS1 region amplicons and Targeted Metabolomics, we analyzed the microbial community structure of fecal microorganisms and Short Chain Fatty Acids (SCFAs) in serum of the Xinjiang fine-wool sheep. Fecal microbial sequencing showed that the bacterial composition and structure were similar between the two groups, whereas there were significant differences in the composition and structure of the fungal community. It was also found that the abundant of Neocallimastigomycota in the intestinal fungal community of FWS was higher. In addition, the results of the serum SCFAs content analysis showed that butyric acid was significantly differences than those two groups. Correlation analysis between SCFAs and bacteria found that butyric acid metabolism had positively correlated (P < 0.05) with Ruminococcus and UCG-005. Overall, our data provide more supplement about the gut microbes community composition and structure of the Xinjiang fine-wool sheep. These results might be useful for improving gut health of sheep and taking nutritional control measure to improve production traits of animals in future.

Faba bean in crop rotation shapes bacterial and fungal communities and nutrient contents under conventional tillage of triticale

Legumes in cropping sequence can strongly moderate soil biodiversity and its biochemical status, which could influence soil fertility and productivity. However, their impact on soil microbes and their relationships with soil chemical variables during the subsequent crop growth is not understood enough. In this study, we analyzed the changes in bacterial and fungal communities structure through 16S rRNA and ITS amplicon sequencing, respectively, across crop rotation with faba bean (faba bean-wheat-triticale, F) and without faba bean (wheat-wheat-triticale, W). Rhizosphere and bulk soil samples were taken during the triticale growth stages (stem elongation and maturity). Soil enzymatic activity and chemical properties were also determined. Study factors (crop rotation, soil compartment, growth stage) affected N, C, and P transformations, as indicated by the activity of soil enzymes, and F was more beneficial than W for protease, urease, and acid phosphomonoesterase activities, as opposed to cellulase activity. Changes in the chemical properties led to the shift in soil microbial communities with different bacterial and fungal communities' responses. F treatment enhanced the abundance of the bacterial genera representatives such as Streptomyces and Candidatus Udaeobacter while suppressing the abundance of Jatrophihabitans and Terrabacter. Crop rotation significantly influenced fungal genera and a greater abundance of Helgardia, Pseudogymnoascus, Monocillium, Fusarium, Chaetomium, and Vishniacozyma under F than W was noted and the adverse situation was noted for Peziza and Rhizopus. Rotation type significantly affected the alfa-diversity of the fungal, but not bacterial community. Beta-diversity analyses (nMDS, cluster) indicated that the main factor grouping samples were soil compartment and growth stage for the bacterial and fungal microbiome, respectively. The PERMANOVA results revealed significant effects of all factors on bacterial and fungal microbiomes. Different soil chemical variables governed bacterial and fungal communities. Corg, pH, P, Mg, and Corg, pH were the most important soil factors regulating bacterial and fungal community structure by crop rotation, respectively. Bacterial and fungal communities were more related to the content of Ntot in rotation with than without faba bean. The findings of this study contribute to a deeper insight into the relation between the faba bean in cropping sequence and soil microbiome, and modulation crucial soil conditions for the productivity of the successive crop.

The maternal microbiome in normal pregnancy and at delivery by cesarean section and the early developmental phase of the neonatal microbiome—presentation of a longitudinal pilot study

Abstract Aim In this study by Foessleitner et al., both the maternal microbiome in the third trimester of pregnancy and the factors that influence the development of the child’s microbiome after cesarean delivery were investigated. Methods Maternal vaginal and rectal swabs were collected at inclusion in the last trimester of pregnancy and on the day of the cesarean section. In addition, placental and intrauterine swabs as well as infant dermal, buccal, and meconium swabs were taken during the cesarean section immediately after birth and subsequently on the second/third day of life. All samples were analyzed for microbial composition using 16s rRNA amplicon sequencing. Results A total of 30 mothers and their newborns were included in the study, with microbiome samples available for all maternal, intrauterine cavity, and placenta samples, as well as for 18 out of the 30 newborns. The vaginal and rectal microbiome was stable over the course of the third trimester and showed no significant changes (permutational multivariate analysis of variance [PERMANOVA]; p &gt; 0.05). Both the intraoperative samples (placental, intrauterine) and the neonatal swabs at the time of birth were consistently sterile. However, rapid infant microbial colonization subsequently occurred, with neonatal buccal mucosa and stool samples showing significantly different microbial colonization from their mothers as early as the second/third day of life (PERMANOVA; p &lt; 0.01). Conclusion The conclusion of the presented study was therefore that the vaginal and rectal microbiome of healthy pregnant women does not change in the last trimester, the infant and the placenta are not microbially colonized at the time of birth, and the development of the newborn’s microbiome after birth appears to be influenced mainly by environmental exposure.

Thermotogota diversity and distribution patterns revealed in Auka and JaichMaa 'ja 'ag hydrothermal vent fields in the Pescadero Basin, Gulf of California.

Discovering new deep hydrothermal vent systems is one of the biggest challenges in ocean exploration. They are a unique window to elucidate the physical, geochemical, and biological processes that occur on the seafloor and are involved in the evolution of life on Earth. In this study, we present a molecular analysis of the microbial composition within the newly discovered hydrothermal vent field, JaichMaa 'ja 'ag, situated in the Southern Pescadero Basin within the Gulf of California. During the cruise expedition FK181031 in 2018, 33 sediment cores were collected from various sites within the Pescadero vent fields and processed for 16S rRNA amplicon sequence variants (ASVs) and geochemical analysis. Correlative analysis of the chemical composition of hydrothermal pore fluids and microbial abundances identified several sediment-associated phyla, including Thermotogota, that appear to be enriched in sediment horizons impacted by hydrothermal fluid flow. Comparative analysis of Thermotogota with the previously explored Auka hydrothermal vent field situated 2 km away displayed broad similarity between the two locations, although at finer scales (e.g., ASV level), there were notable differences that point to core-to-core and site-level factors revealing distinct patterns of distribution and abundance within these two sediment-hosted hydrothermal vent fields. These patterns are intricately linked to the specific physical and geochemical conditions defining each vent, illuminating the complexity of this unique deep ocean chemosynthetic ecosystem.

A finding of potential coexisting bacteria and characterization of the bacterial communities in the fruiting body of Sarcodon aspratus.

Sarcodon aspratus (Berk.) S. Ito is a Japaneselocal dish with unique aroma and is effective against allergic diseases. However, its cultivation was still difficult. Recently, coexisting bacteria were regarded as an important factor for mycelium growth and fruiting body formation. Therefore, we performed 16S rRNA amplicon sequencing in the fruiting body of S. aspratus and its adhered soil to understand the bacterial communities in the fruiting body of S. aspratus. The fruiting body group showed lower alpha diversities and a significant difference in the structure of bacterial communities compared to the soil group. In addition, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium had the highest relative abundance in the fruiting body group, and it was also a potential coexisting bacterium in the fruiting body of S. aspratus by linear discriminant analysis effect size (LEfSe) analysis. This highest relative abundance phenomenon in Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium clade was also found in the fruiting body of Cantharellus cibarius. These findings suggested that Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium plays a key role in the bacterial communities in the fruiting body of S. aspratus. Bacteria in the fruit bodies of S. aspratus and C. cibarius probably present a similar coexistence model.

Specific Gut Microbiome Signatures in Children with Cow's Milk Allergy.

Although gut dysbiosis is associated with cow's milk allergy (CMA), causality remains uncertain. This study aimed to identify specific bacterial signatures that influence the development and outcome of the disease. We also investigated the effect of hypoallergenic formula (HF) consumption on the gut microbiome of milk-allergic children. 16S rRNA amplicon sequencing was applied to characterize the gut microbiome of 32 milk-allergic children aged 5-12 years and 36 age-matched healthy controls. We showed that the gut microbiome of children with CMA differed significantly from that of healthy children, regardless of whether they consumed cow's milk. Compared to that of healthy cow's milk consumers, it was depleted in Bifidobacterium, Coprococcus catus, Monoglobus, and Lachnospiraceae GCA-900066575, while being enriched in Oscillibacter valericigenes, Negativibacillus massiliensis, and three genera of the Ruminococcaceae family. Of these, only the Ruminococcaceae taxa were also enriched in healthy children not consuming cow's milk. Furthermore, the gut microbiome of children who developed tolerance and had received an HF was similar to that of healthy children, whereas that of children who had not received an HF was significantly different. Our results demonstrate that specific gut microbiome signatures are associated with CMA, which differ from those of dietary milk elimination. Moreover, HF consumption affects the gut microbiome of children who develop tolerance.