Metagenomic Analysis Research Articles

Metagenome Analysis of the Bacterial Communities of mangrove sediments from Neil Island of Andamans

Metagenome Analysis of the Bacterial Communities of mangrove sediments from Neil Island of Andamans

Metabolic-methane mitigation by combination of probiotic Escherichia coli strain Nissle 1917 and biochar in rumen fluid in vitro fermentation of dairy cow

In response to climate change, there have been various trials to reduce methane emissions from ruminant animals in the livestock industry, and one of the typical candidates is probiotics used as feed additives. We employed Escherichia coli Nissle 1917 (EcN) as a promising probiotic for the reduction of methane emissions and investigated the molecular mechanism of methane-reducing effects using CRISPR/Cas9. The anaerobic culture of EcN with biochar (BC) could significantly increase acetate and hydrogen production. Additionally, RNA-seq analysis revealed an upregulation in the expression of acetate synthesis genes, namely, pta, poxB, and ackA. Subsequently, treatment of rumen fluid with EcN significantly decreased methane and increased acetate and propionate production, as observed through the analysis of short-chain fatty acids, and these effects could accelerate with additional supplementation of BC. The observation of changes in the microbial composition of rumen fluid following treatment with EcN and BC was made through rumen metagenomic analysis and RT-qPCR. The results revealed an increase in acetogen and propionate producing bacteria abundance and a decrease in methanogen abundance. Based on these findings, the CRISPR/Cas9 system was employed to elucidate the molecular mechanism of the methane-reducing capability of EcN. Gene deletion was performed targeting the genes poxB, ackA, and pta as key factors in acetate pathway in E. coli. The knockout of poxB, ackA, and pta could lead to the elimination of the methane-reduction of EcN as well as acetate-producing abilities. Collectively, the methane reduction ability of EcN in rumen fluid is associated with its acetate production capability, and the addition of BC significantly enhances methane mitigation capability of EcN.

Molecular Analysis of Indole and Skatole Decomposition Metabolism in Acinetobacter piscicola p38 Utilizing Biochemical and Omics Approaches.

Indole and skatole (3-methylindole, C9H9N) are common nitrogen-containing heterocyclic pollutants found in waste, wastewater treatment plants, and public restrooms and are the most notorious compounds in animal feces. Biodegradation was considered a feasible method for the removal of indole and skatole, but a comprehensive understanding of the metabolic pathways under both aerobic and anaerobic conditions was lacking, and the functional genes responsible for skatole biodegradation remained a mystery. Through metagenomic and gene cluster functional analysis, Acinetobacter piscicola p38 (NCBI: CP167896), genes 1650 (styrene monooxygenase: ACDW34_08180), and 1687 (styrene monooxygenase: ACDW34_08350) were identified as having the potential to degrade indole and skatole. The heterologous expression results demonstrate that the genes 1650 and 1651 (flavin reductase: ACDW34_08185), when combined, are capable of degrading indole, while the genes 1687 and 1688 (flavin reductase: ACDW34_08355), in combination, can degrade indole as well as skatole. These reactions necessitate the involvement of flavin reductase and NAD(P)H to catalyze the oxygenation process. This work aimed to provide new experimental evidence for the biodegradation of indole and skatole. This study offered new insights into our understanding of skatole degradation. The Acinetobacter_piscicola p38 strain provided an effective bacterial resource for the bioremediation of fecal indole and skatole.

Methanogenesis coupled hydrocarbon biodegradation enhanced by ferric and sulphate ions

Bioremediation provides an environmentally sound solution for hydrocarbon removal. Although bioremediation under anoxic conditions is slow, it can be coupled with methanogenesis and is suitable for energy recovery. By altering conditions and supplementing alternative terminal electron acceptors to the system to induce syntrophic partners of the methanogens, this process can be enhanced. In this study, we investigated a hydrocarbon-degrading microbial community derived from chronically contaminated soil. Various hydrocarbon mixtures were used during our experiments in the presence of different electron acceptors. In addition, we performed whole metagenome sequencing to identify the main actors of hydrocarbon biodegradation in the samples. Our results showed that the addition of ferric ions or sulphate increased the methane yield. Furthermore, the addition of CO2, ferric ion or sulphate enhanced the biodegradation of alkanes. A significant increase in biodegradation was observed in the presence of ferric ions or sulphate in the case of all aromatic components, while naphthalene and phenanthrene degradation was also enhanced by CO2. Metagenome analysis revealed that Cellulomonas sp. is the most abundant in the presence of alkanes, while Ruminococcus and Faecalibacterium spp. are prevalent in aromatics-supplemented samples. From the recovery of 25 genomes, it was concluded that the main pathway of hydrocarbon activation was fumarate addition in both Cellulomonas, Ruminococcus and Faecalibacterium. Chloroflexota bacteria can utilise the central metabolites of aromatics biodegradation via ATP-independent benzoyl-CoA reduction.Key points• Methanogenesis and hydrocarbon biodegradation were enhanced by Fe3+ or SO42−• Cellulomonas, Ruminococcus and Faecalibacterium can be candidates for the main hydrocarbon degraders• Chloroflexota bacteria can utilise the central metabolites of aromatics degradationGraphical

Domestication of halotolerant methanogens for energy-efficient microbial electrosynthesis of methane from CO2

Microbial electrosynthesis (MES) has great potential for CO2 reduction and renewable energy storage. However, high energy consumption and investment costs constrained the practical application of MES. Here, the domestication of halotolerant methanogens was proposed to improve the energy efficiency of MES, and methane metabolism under low and high Na+ concentrations was explored by metagenomic analysis. The Na+ concentration of the catholyte was increased from 4.07 g·L-1 to 23.01 g·L-1, and the reactor maintained great performance after initial inhibition. High-salinity catholyte improved the energy efficiency to 47% and reduced the cathode voltage consumption. Increasing Na+ concentration, microorganisms resisted damage by secreting more extracellular polymers, and maintained methane production through community evolution. The relative abundance of Euryarchaeota increased from 50% to 80%. Methanobrevibacter and Methanobacterium were dominant genera at Na+ of 23.01 g·L-1. Incorporating metagenomic analysis, the methane metabolism at the initial period (Na+:4.07 g·L-1) included a CO2-reduction pathway and acetoclastic pathway. However, when Na+ concentration increased to 23.01 g·L-1, the CO2-reduction pathway with 4Fe–4S ferredoxin and common pathways was significantly enhanced, in addition, the acetyl metabolism pathway using CO2 as substrate was also enhanced. Overall, methane metabolism was enhanced, and the metabolic pathways changed with increasing Na⁺ concentration.

Effect of Organic and Chemical Fertilizer on the Diversity of Rhizosphere and Leaf Microbial Composition in Sunflower Plant.

Applying organic manure to crops positively impacts the soil microbial community which is negatively impacted when chemical fertilizers are used. Organic manures also add new microbes to the soil in addition to influencing the growth of native ones. Metagenomic analysis of different organic manures, soil, and pot culture experiments conducted under various fertilizer conditions constitute the primary methodologies employed in this study. We compared the effect of two organic manure combinations and an inorganic fertilizer combination on microbial community of rhizosphere soil and leaves of sunflower plants. Metagenomic sequencing data analysis revealed that the diversity of bacteria and fungi is higher in organic manure than in chemical fertilizers. Each organic manure combination selectively increased population of some specific microbes and supported new microbes. Application of chemical fertilizer hurts many plant beneficial fungi and bacteria. In summary, our study points out the superiority of organic manure combinations in enhancing microbial diversity and supporting beneficial microbes. These findings enhance the profound influence of fertilizer types on sunflower microbial communities, shedding light on the intricate dynamics within the rhizosphere and leaf microbiome. Bacterial genera such as Bacillus, Serratia, Sphingomonas, Pseudomonas, Methylobacterium, Acinetobacter, Stenotrophomonas, and fungal genera such as Wallemia, Aspergillus, Cladosporium, and Penicillium constitute the key microbes of sunflower plants.

Linking feed, biodiversity, and filtration performance in a Self-Forming Dynamic Membrane BioReactor (SFD MBR) treating canning wastewater

A bench-scale self-forming dynamic membrane bioreactor (SFD MBR) treating canning wastewater showed instable filtration performance, with a worsening in correspondence of a change in the influent quality. To unveil the possible reasons for such behaviour, a microbial investigation was carried out to complement conventional filterability and settleability tests. The microbial communities of the influent wastewater, the activated sludge, and the filtering sludge cake (dynamic membrane) were characterised through metagenomic and differential abundance analyses. In parallel, the production of exopolymers was measured. The outputs indicate that under the tested conditions the canning wastewater promoted the development of a very peculiar microbial ecosystem, with a strong selection of the genus Thiothrix (relative abundance above 90 %). The Shannon index values in activated sludge were below 2, indicating a very low bacterial diversity. This may have limited the ecosystem resilience towards changes of external conditions, causing instability in the process of the dynamic membrane formation. Furthermore, the diversity analysis showed that the bacterial compositions of the feed and the sludge resulted not significantly linked to each other, confirming the main role of the chemical composition of the influent wastewater on overall process performances. When treating agro-industrial streams, in-depth feed characterisation is recommended in order to prevent the factors that may generate instability in SFD MBR performance.

Metagenomic mining of two Egyptian Red Sea sponges associated microbial community

The Red Sea is a promising habitat for the discovery of new bioactive marine natural products. Sponges associated microorganisms represent a wealthy source of compounds with unique chemical structures and diverse biological activities. Metagenomics is an important omics-based culture-independent technique that is used as an effective tool to get genomic and functional information on sponge symbionts. In this study, we used metagenomic analysis of two Egyptian Red Sea sponges Hyrtios erectus and Phorbas topsenti microbiomes to study the biodiversity and the biosynthetic potential of the Red Sea sponges to produce bioactive compounds. Our data revealed high biodiversity of the two sponges’ microbiota with phylum Proteobacteria as the most dominant phylum in the associated microbial community with an average of 31% and 70% respectively. The analysis also revealed high biosynthetic potential of sponge Hyrtios erectus microbiome through detecting diverse types of biosynthetic gene clusters (BGCs) with predicted cytotoxic, antibacterial and inhibitory action. Most of these BGCs were predicted to be novel as they did not show any similarity with any MIBiG database known cluster. This study highlights the importance of the microbiome of the collected Red Sea sponge Hyrtios erectus as a valuable source of new bioactive natural products.

Metagenomic Analysis of Bottom Sediments of the Karst Meromictic Lake Black Kichier Revealed Abundant Unculturable Thermoplasmatota

Investigations into the microbial community of bottom sediments in the karst lake Black Kichier were conducted. These sediments exhibited elevated levels of sulfide, dissolved methane, and organic matter. Direct radiotracer experiments revealed substantial rates of microbial processes involved in the decomposition of organic matter. Uncultivated archaea belonging to the phylum Thermoplasmatota were identified within the microbial community. Metagenomic analysis unveiled representatives from five orders: Methanomassiliicoccales, Thermoprofundales (formerly known as Marine Benthic Group D and DHVEG-1), DTX01, SG8-5, and Candidatus Gimiplasmatales (formerly UBA10834). These archaea were previously believed to occur exclusively in deep marine sediments characterized by extreme organic matter scarcity. This discovery reshapes our understanding of the role played by Thermoplasmatota archaea, spanning five orders, in the degradation segment of the carbon cycle.

Consistent signatures in the human gut microbiome of longevous populations

ABSTRACT Gut microbiota of centenarians has garnered significant attention in recent years, with most studies concentrating on the analysis of microbial composition. However, there is still limited knowledge regarding the consistent signatures of specific species and their biological functions, as well as the potential causal relationship between gut microbiota and longevity. To address this, we performed the fecal metagenomic analysis of eight longevous populations at the species and functional level, and employed the Mendelian randomization (MR) analysis to infer the causal associations between microbial taxa and longevity-related traits. We observed that several species including Eisenbergiella tayi, Methanobrevibacter smithii, Hungatella hathewayi, and Desulfovibrio fairfieldensis were consistently enriched in the gut microbiota of long-lived individuals compared to younger elderly and young adults across multiple cohorts. Analysis of microbial pathways and enzymes indicated that E. tayi plays a role in the protein N-glycosylation, while M. smithii is involved in the 3-dehydroquinate and chorismate biosynthesis. Furthermore, H. hathewayi makes a distinct contribution to the purine nucleobase degradation I pathway, potentially assisting the elderly in maintaining purine homeostasis. D. fairfieldensis contributes to the menaquinone (vitamin K2) biosynthesis, which may help prevent age-related diseases such as osteoporosis-induced fractures. According to MR results, Hungatella was significantly positively correlated with parental longevity, and Desulfovibrio also exhibited positive associations with lifespan and multiple traits related to parental longevity. Additionally, Alistipes and Akkermansia muciniphila were consistently enriched in the gut microbiota of the three largest cohorts of long-lived individuals, and MR analysis also suggests their potential causal relationships with longevity. Our findings reveal longevity-associated gut microbial signatures, which are informative for understanding the role of microbiota in regulating longevity and aging.

Relationship between microorganisms and milk metabolites during quality changes in refrigerated raw milk: A metagenomic and metabolomic exploration

Although cold storage at 4 °C can effectively prolong the shelf life of raw milk, it cannot prevent its eventual spoilage. In this study, we analyzed the main physicochemical and microbial indexes of raw milk stored at 4 °C for 6 days. The changes in microbial profiles and milk metabolites and their relationship during refrigeration were also explored. Metagenomic analysis performed using the Illumina Hiseq Xten sequencing platform revealed that the dominant genera in raw milk evolved from Acinetobacter, Streptococcus, Staphylococcus, and Anaplasma to Flavobacterium, Pseudomonas, and Lactococcus during cold storage. Using the UHPLC-Q-TOF MS method, 77 significantly different metabolites (p < 0.05) were identified, among which lipids were the most abundant (37). The most significant metabolic changes largely occurred at 3–4 days of refrigeration, coinciding with the rapid increase in dominant psychrotrophic bacteria. Subsequently, correlation analysis demonstrated that these lipid-related metabolites were significantly associated with Acinetobacter, Flavobacterium, and Pseudomonas. Both macro indicators and microanalysis indicated that the key stage of quality changes in raw milk was 3–4 days. Thus, this stage can be targeted for the quality control of raw milk.

Shotgun metagenomic analysis reveals the emergence of plasmid-encoded mcr-5.1 gene in hospital wastewater in Bangladesh

Colistin is considered the last line therapy for treating multidrug-resistant (MDR) bacterial infections in humans. Therefore, the spread of colistin resistance poses a serious threat to human, and environmental health. Though Bangladesh is known as a hotspot of AMR, limited studies have been carried out regarding the status of colistin resistance. Information on the emerging bacterial resistance is inevitable for protecting public health. Nowadays, wastewater analysis has been prioritized for metagenomics-enabled AMR surveillance. Our study on the metagenomic analysis of untreated hospital effluents first detected the colistin resistance-conferring mcr-5.1 gene in the hospital environment of Bangladesh. Phylogenetic tree and in silico AMR analysis confirmed the detection of this mcr-5 variant, which is located in a plasmid contig. The plasmid was untypeable and belonged to the bacteria from the Enterobacteriaceae family. The mcr-5.1 operon was embedded in a Tn3 transposon, suggesting the mobility of the gene. Tnshfr1 transposon, chromate resistance protein ChrB, DNA invertase hin, and two MFS-type proteins were present in the genetic environment of mcr-5.1. Our findings provide evidence of the occurrence of mcr-5.1 in a hospital environment in Bangladesh, which calls for immediate attention and effective measures to contain the dissemination of colistin resistance in the environment.

"Candidatus Uabimicrobium helgolandensis"-a planctomycetal bacterium with phagocytosis-like prey cell engulfment, surface-dependent motility, and cell division.

The unique cell biology presented by members of the phylum Planctomycetota has puzzled researchers ever since their discovery. Initially thought to have eukaryotic-like features, their traits are now recognized as exceptional but distinctly bacterial. However, recently discovered strains again added novel and stunning aspects to the planctomycetal cell biology-shapeshifting by members of the "Saltatorellus" clade to an extent that is unprecedented in any other bacterial phylum, and phagocytosis-like cell engulfment in the bacterium "Candidatus Uabimicrobium amorphum." These recent additions to the phylum Planctomycetota indicate hitherto unexplored members with unique cell biology, which we aimed to make accessible for further investigations. Targeting bacteria with features like "Ca. U. amorphum", we first studied both the morphology and behavior of this microorganism in more detail. While similar to eukaryotic amoeboid organisms at first sight, we found "Ca. U. amorphum" to be rather distinct in many regards. Presenting a detailed description of "Ca. U. amorphum," we furthermore found this organism to divide in a fashion that has never been described in any other organism. Employing the obtained knowledge, we isolated a second "bacterium of prey" from the harbor of Heligoland Island (North Sea, Germany). Our isolate shares key features with "Ca. U. amorphum": phagocytosis-like cell engulfment, surface-dependent motility, and the same novel mode of cell division. Being related to "Ca. U. amorphum" within genus thresholds, we propose the name "Ca. Uabimicrobium helgolandensis" for this strain.IMPORTANCE"Candidatus Uabimicrobium helgolandensis" HlEnr_7 adds to the explored bacterial biodiversity with its phagocytosis-like uptake of prey bacteria. Enrichment of this strain indicates that there might be "impossible" microbes out there, missed by metagenomic analyses. Such organisms have the potential to challenge our understanding of nature. For example, the origin of eukaryotes remains enigmatic, with a contentious debate surrounding both the mitochondrial host entity and the moment of uptake. Currently, favored models involve a proteobacterium as the mitochondrial progenitor and an Asgard archaeon as the fusion partner. Models in which a eukaryotic ancestor engulfed the mitochondrial ancestor via phagocytosis had been largely rejected due to bioenergetic constraints. Thus, the phagocytosis-like abilities of planctomycetal bacteria might influence the debate, demonstrating that prey engulfment is possible in a prokaryotic cellular framework.

Effect of symbiotic microbiota and arginine on host plant selection by the tea green leafhopper in tea cultivation

The tea green leafhopper (TGLH), Empoasca (Matsumurasca) onukii Matsuda, poses a significant challenge to tea cultivation in East Asia because of its ability to infest specific tea plant cultivars. Here, we investigated the complex interactions between TGLHs, various tea cultivars, and their symbiotic microbiota, especially the effect of arginine on host selection. A detailed two-year field study involving 16 tea cultivars, coupled with controlled laboratory and field experiments, was conducted to establish a clear link between TGLH infestation and the arginine content of tea plants. Metagenomic analysis revealed a predominance of bacterial taxa in TGLHs, primarily from the phylum Proteobacteria, with significant contributions from orders such as Pseudomonadales, Enterobacterales, and Flavobacteriales, which are crucial for the biosynthesis of essential amino acids but lack the argF gene necessary for arginine synthesis. The absence of the argF gene indicates the potential presence of microbial-mediated dietary adaptation strategies in TGLHs. Additionally, our research examines microbial diversity within TGLH populations and confirms the critical role that symbiotic bacteria play in influencing dietary preferences. The results of our study provide insights into the dynamic interplay between pests, plants, and microorganisms. Our findings are preliminary, and further research is needed to explore the potential applications of symbiotic bacteria in sustainable tea cultivation and integrated pest management strategies.

Sewerage surveillance tracking characteristics of human antibiotic resistance genes in sewer system

Sewage surveillance is widely applied to track valid human excretion information and identify public health conditions during corona virus disease 2019 (COVID-19) pandemic. This approach can be applied to monitor the antibiotic resistance level in sewers and to assess the risk of spreading antibiotic resistance in municipal wastewater systems. However, there is still little information about human antibiotic resistance occurrence characteristics in sewer system. This study conducted a field trial for whole year to advance understanding on spatial and temporal occurrence of antibiotic resistance genes (ARGs) in gravity sewerage. The spatial distribution of ARGs along the drainage pipe line (from human settlements to wastewater treatement pant (WWTP)) was insignificant, which may be affected by irregular human emission alongside the pipeline. The correlation between ARGs and antibiotics in sewage was insignificant. The temporal distribution showed that the effect of temperature on ARGs abundance was evident, the ARGs abundance in sewage was generally higher during the cold season. Metagenomic analysis revealed that the detected ARGs were mainly distributed in Proteobacteria (47.51 %) and Antinobacteria (20.11 %). Potential hosts of ARGs in sewage were mainly identified as human gut microorganisms, including human pathogenic bacteria, such as Prevotella, Kocuria, and Propionibacterium, etc. This study provides a new insight into the sewerage surveillance tracking characteristics of human ARGs in sewer system, and suggesting that the sewage-carried ARGs surveillance is a promising method for assessment and management of antibiotic resistance level on population size.

Enhancing the performance of microalgal-bacterial systems with sodium bicarbonate: A step forward to carbon neutrality of municipal wastewater treatment

The microalgal-bacterial granular sludge (MBGS) process, enhanced with sodium bicarbonate (NaHCO3), offers a sustainable alternative for wastewater treatment aiming for carbon neutrality. This study demonstrates that NaHCO3, which can be derived from the flue gases and alkaline textile wastewater, significantly enhances pollutant removal and biomass production. Optimal addition of NaHCO3 was found to achieve an inorganic-to-organic carbon ratio of 1.0 and a total carbon-to-nitrogen ratio of 5.0. Metagenomic analysis and structural equation modeling showed that NaHCO3 addition increased dissolved oxygen concentrations and pH levels, creating a more favorable environment for key microbial communities, including Proteobacteria, Chloroflexi, and Cyanobacteria. Confocal laser scanning microscopy further confirmed enhanced interactions between Cyanobacteria and Proteobacteria/Chloroflexi, facilitating the MBGS process. These microbes harbored functional genes (gap2, GLU, and ppk) critical for removing organics, nitrogen, and phosphorus. Carbon footprint analysis revealed significant reductions in CO2 emissions by the NaHCO3-added MBGS process in representative countries (China, Australia, Canada, Germany, and Morocco), compared to the conventional activated sludge process. These findings highlight the effectiveness of NaHCO3 in optimizing MBGS process, establishing it as a key strategy in achieving carbon-neutral wastewater treatment globally.

Metagenomic analyses of aerobic bacterial enrichment cultures that degraded Tris(2-chloroethyl) phosphate (TCEP) and its transformation products

Tris(2-chloroethyl) phosphate (TCEP) is of growing public concern worldwide because of its ubiquitous contamination, toxicity, and persistence. In this study, we investigated bacterial communities in aerobic enrichment cultures with TCEP and its two major transformation products bis(2-chloroethyl) phosphate (BCEP) and 2-chloroethanol (2-CE) as the sole carbon source. Burkholderiales and Rhizobiales were likely two main bacterial guilds involved in the hydrolysis of TCEP, while Rhizobiales and Sphingomonadales may play an important role in the hydrolysis of BCEP, given the increase of Rhizobiales and Sphingomonadales-related phosphoesterase genes when the carbon source was switched from TCEP to BCEP. All Burkholderiales, Rhizobiales, Sphingomonadales were probably efficient in 2-CE metabolism, because their dehydrogenase genes and dehalogenase genes increased after 2-CE enrichment. The different substrate preference of different bacterial guilds highlighted the importance to understand the diversity and collaboration among functional bacteria. Meanwhile, two TCEP-degrading strains affiliated with Xanthobacter and Ancylobacter were isolated. Xanthobacter sp. strain T2-1 was able to degrade both TCEP and BCEP following the pseudo-first-order kinetics with reaction rates of 1.66 h−1 for TCEP and 1.02 h−1 for BCEP, respectively. Ancylobacter sp. strain T3-4 could degrade TCEP following the pseudo-first-order kinetics with a reaction rate of 2.54 h−1, but is unable to degrade BCEP. Additionally, strains that were phylogenetically closely related were found to have different degradation capabilities toward TCEP and/or BCEP, indicating the importance to investigate functional genes such as phosphoesterase genes.

Metagenomics analysis of sewage for surveillance of antimicrobial resistance in South Africa.

Our 24-month study used metagenomics to investigate antimicrobial resistance (AMR) abundance in raw sewage from wastewater treatment works (WWTWs) in two municipalities in Gauteng Province, South Africa. At the AMR class level, data showed similar trends at all WWTWs, showing that aminoglycoside, beta-lactam, sulfonamide and tetracycline resistance was most abundant. AMR abundance differences were shown between municipalities, where Tshwane Metropolitan Municipality (TMM) WWTWs showed overall higher abundance of AMR compared to Ekurhuleni Metropolitan Municipality (EMM) WWTWs. Also, within each municipality, there were differing trends in AMR abundance. Notably, within TMM, certain AMR classes (macrolides and macrolides_streptogramin B) were in higher abundance at a WWTW serving an urban high-income area, while other AMR classes (aminoglycosides) were in higher abundance at a WWTW serving a semi-urban low income area. At the AMR gene level, all WWTWs samples showed the most abundance for the sul1 gene (encoding sulfonamide resistance). Following this, the next 14 most abundant genes encoded resistance to sulfonamides, aminoglycosides, macrolides, tetracyclines and beta-lactams. Notably, within TMM, some macrolide-encoding resistance genes (mefC, msrE, mphG and mphE) were in highest abundance at a WWTW serving an urban high-income area; while sul1, sul2 and tetC genes were in highest abundance at a WWTW serving a semi-urban low income area. Differential abundance analysis of AMR genes at WWTWs, following stratification of data by season, showed some notable variance in six AMR genes, of which blaKPC-2 and blaKPC-34 genes showed the highest prevalence of seasonal abundance differences when comparing data within a WWTW. The general trend was to see higher abundances of AMR genes in colder seasons, when comparing seasonal data within a WWTW. Our study investigated wastewater samples in only one province of South Africa, from WWTWs located within close proximity to one another. We would require a more widespread investigation at WWTWs distributed across all regions/provinces of South Africa, in order to describe a more comprehensive profile of AMR abundance across the country.

Lactobacillus plantarum Ameliorates Colorectal Cancer by Ameliorating the Intestinal Barrier through the CLA-PPAR-γ Axis.

Colorectal cancer (CRC) is the third-largest cancer worldwide. Lactobacillus can regulate the intestinal barrier and gut microbiota. However, the mechanisms of Lactobacillus that alleviate CRC remained unknown. This study aimed to explore the regulatory effect of Lactobacillus plantarum on CRC and its potential mechanism. CCFM8661 treatment significantly ameliorated CRC compared with phosphate-buffered solution (PBS) treatment in ApcMin/+ mice. In addition, conjugated linoleic acid (CLA) was proved to be the key metabolite for CCFM8661 in ameliorating CRC by molecular biology techniques. Peroxisome proliferator-activated receptor γ (PPAR-γ) was proved to be the key receptor in ameliorating CRC by inhibitor intervention experiments. Moreover, supplementation with CCFM8661 ameliorated CRC by producing CLA to inhibit NF-κB pathway and pro-inflammatory cytokines, up-regulate ZO-1, Claudin-1, and MUC2, and promote tumor cell apoptosis in a PPAR-γ-dependent manner. Metagenomic analysis showed that CCFM8661 treatment significantly increased Odoribacter splanchnicus, which could ameliorate CRC by repairing the intestinal barrier. Clinical results showed that intestinal CLA, butyric acid, PPAR-γ, and Lactobacillus were significantly decreased in CRC patients, and these indicators were significantly negatively correlated with CRC. CCFM8661 alleviated CRC by ameliorating the intestinal barrier through the CLA-PPAR-γ axis. These results will promote the development of dietary probiotic supplements for CRC.

Amugulang virus, a novel hantavirus harboured by small rodents in Hulunbuir, China

ABSTRACT The Hulunbuir region, known for its diverse terrain and rich wildlife, is a hotspot for various natural epidemic diseases. Between 2021 and 2023, we collected 885 wild rodent samples from this area, representing three families, seven genera, and eleven species. Metagenomic analysis identified three complete nucleic acid sequences from the S, M, and L segments of the Hantaviridae family, which were closely related to the Khabarovsk virus. The nucleotide coding sequences for S, M, and L (1392 nt, 3465 nt, and 6491 nt, respectively) exhibited similarities of 82.34%, 81.68%, and 81.94% to known sequences, respectively, while protein-level analysis indicated higher similarities of 94.92%, 94.41%, and 95.87%, respectively. Phylogenetic analysis placed these sequences within the same clade as the Khabarovsk, Puumala, Muju, Hokkaido, Topografov, and Tatenalense viruses, all of which are known to cause febrile diseases in humans. Immunofluorescence detection of nucleic acid-positive rodent kidney samples using sera from patients with hemorrhagic fever and renal syndrome confirmed the presence of viral particles. Based on these findings, we propose that this virus represents a new member of the Hantaviridae family, tentatively named the Amugulang virus, after its primary distribution area.