16S rRNA Gene Sequence Research Articles

Deciphering the microbial communities in ticks of Inner Mongolia: ecological determinants and pathogen profiles

BackgroundTicks are vectors of numerous pathogens, with their bacterial composition, abundance, diversity, and interaction influencing both their growth and disease transmission efficiency. Despite the abundance of ticks in Inner Mongolia, China, comprehensive data on their microbial communities are lacking. This study aims to analyze the microbial communities within ticks from Inner Mongolia to inform innovative control strategies for interrupting pathogen transmission.MethodsTick samples were collected from animals and vegetation in multiple locations across Inner Mongolia and stored at − 80 °C. Ticks were identified using morphological keys and molecular biology methods. Full-length 16S rRNA gene sequencing was performed on collected samples. Bacterial community composition and diversity were mainly analyzed using bioinformatic tools such as QIIME, phyloseq, and DESeq2. Alpha diversity was assessed using Chao1, ACE, and Shannon indices, while beta diversity was evaluated using Bray-Curtis dissimilarity matrices. LEfSe analysis was applied to identify taxa associated with ecological and biological variables.ResultsA total of 5,048,137 high-quality read counts were obtained, forming an average of 789.3 OTUs per sample. Proteobacteria, Firmicutes, and Bacteroidetes were the most dominant phyla. Bacterial community composition varied significantly with geography, with Dermacentor nuttalli showing a higher abundance of Rickettsia in Xilingol League, while other regions had different dominant genera. The microbial community also differed based on the feeding status of ticks. Additionally, the microbiota of engorged ticks showed organ specificity. Pathogen detection efforts revealed the presence of nine pathogens across all three tick species. D. nuttalli was found to carry a significantly higher burden of pathogenic bacteria, making it the most potentially threatening tick species in Inner Mongolia.ConclusionsThe study highlights significant variations in tick microbiomes influenced by geographic location, feeding status, and tick species. It underscores the importance of enhancing tick and tick-borne disease surveillance in Inner Mongolia for early detection and control of emerging pathogens.Graphical

Recurrent association between Trichodesmium colonies and calcifying amoebae

Abstract Colonies of the N2-fixing cyanobacterium Trichodesmium spp. constitute a consortium with multiple microorganisms that collectively exert ecosystem-level influence on marine carbon and nitrogen cycling, shunting newly fixed nitrogen to low nitrogen systems, and exporting both carbon and nitrogen to the deep sea. Here we identify a seasonally recurrent association between puff colonies and amoebae through a two-year survey involving over 10 000 Trichodesmium colonies in the Red Sea. This association was most commonly found in near-shore populations during spring. Microscopic observations revealed consistent amoebae morphology throughout the study, and both morphological characteristics and 18S rRNA gene sequencing suggested that these amoebae are likely to belong to the species Trichosphaerium micrum, an amoeba that forms a CaCO3 shell. Co-cultures of Trichosphaerium micrum and Trichodesmium grown in the laboratory suggest that the amoebae feed on heterotrophic bacteria and not Trichodesmium, which adds a consumer dynamic to the complex microbial interactions within these colonies. Sinking experiments with fresh colonies indicated that the presence of the CaCO3-shelled amoebae decreased colony buoyancy. As such, this novel association may accelerate Trichodesmium sinking rates and facilitate carbon and nitrogen export to the deep ocean. Amoebae have previously been identified in Trichodesmium colonies in the western North Atlantic (Bermuda and Barbados), suggesting that this type of association may be widespread. This association may add a new critical facet to the microbial interactions underpinning carbon and nitrogen fixation and fate in the present and future ocean.

Dietary supplementation with Rehmannia glutinosa catalpol improves broiler performance by regulating intestinal immune microenvironment

Plant extracts have been widely considered as a feed additive. Rehmannia glutinosa catalpol can promote the growth performance of the host, improve the immune response, and strengthen the intestinal barrier function of the host. The broilers were fed with Rehmannia glutinosa catalpol (0.01%), and the composition of caecum microbiota in broilers was explored by 16S rRNA gene sequencing. Finally, the changes of intestinal mucosa morphology and immune function were analysed by tissue staining and RT-qPCR. The proportion of thick-walled bacteria in the treatment group was higher than that in the control group. At the genus level, the relative abundance of Faecalibacterium, Paraacteroides and Barnesiella increased after the addition of Rehmannia glutinosa catalpol. The function prediction results of PICRUSt2 showed that the microbiota mainly participated in the biosynthesis and degradation/utilization/association pathway. The results of tissue staining and RT-qPCR showed that the caecum tissue morphology and immune barrier function of broilers were significantly improved after the addition of catalpol. The results of this study show that Rehmannia glutinosa catalpol in the diet may be beneficial to the intestinal health of broilers, which may be achieved by regulating the intestinal microbiota and enhancing the immune function of broilers.

Impact of Dietary Fatty Acid Composition on the Intestinal Microbiota and Fecal Metabolism of Rats Fed a High-Fructose/High-Fat Diet.

Background/Objectives: An inappropriate intake of dietary fats can disrupt the homeostasis of intestinal microbiota, affect the host's metabolic status, and increase the risk of chronic diseases. The impact of dietary fat types on the composition and metabolic functionality of the intestinal microbiota has become a research focus over recent years. The objective of this study was to explore the effects of regular peanut oil (PO) and high-oleic-acid peanut oil (HOPO) on the composition and metabolic function of the intestinal microbiota. Methods: A dietary intervention test was conducted on SD rats fed a high-fat/high-fructose (HFF) diet. The composition and metabolic functionality of the intestinal microbiota of the experimental rats were investigated by 16S rRNA gene sequencing and fecal metabolomics. Results: Compared with saturated fat, PO and HOPO enhanced the diversity of intestinal microbiota in HFF diet-fed rats. Compared with PO, HOPO significantly increased the relative abundance of Lachnospiraceae_NK4A136_group and Harryflintia (p < 0.05), which are able to generate butyrate and acetate. Compared with saturated fat, 318 and 271 fecal biomarkers were identified in PO and HOPO groups, respectively. In contrast, 68 fecal biomarkers were identified between the PO and HOPO groups. The inhibition of harmful proteolytic fermentation in the colon may represent the main regulatory mechanism. With regard to metabolic status, HOPO provided better control of body weight and insulin sensitivity than PO. Conclusions: Compared with saturated fat, peanut oils better regulated the composition and metabolic function of the intestinal microbiota. In addition, HOPO exhibited better regulatory effects than PO.

Sulfate-driven microbial collaboration for synergistic remediation of chloroethene-heavy metal pollution

The treatment of heavy metal(loid) (HM) composite pollution has long posed a challenge for the bioremediation of organohalide-contaminated sites. Given the prevalent cohabitation of sulfate-reducing bacteria (SRB) with organohalide-respiring bacteria (OHRB), we proposed a sulfate-amendment strategy to achieve synergistic remediation of trichloroethene and diverse HMs [50μM of As(III), Ni(II), Cu(II), Pb(II)]. Correspondingly, 50–75 μM sulfate was introduced to HM inhibitory batches to investigate the enhancement effect of sulfate amendment on bio-dechlorination. Dechlorination kinetics and MATLAB modeling indicated that sulfate amendment comprehensively improved the reductive dechlorination performance in the presence of As(III), Ni(II), Pb(II) and mixed HMs, while no enhancement was observed under Cu(II) exposure. Additionally, sulfate introduction effectively accelerated the detoxification of Ni(II), Pb(II), Cu(II), and As(III), achieving removal efficiencies of 76.87 %, 64.01 %, 86.37 %, and 95.50 % within the first three days, respectively. Meanwhile, propionate dynamics and acetogenesis indicated enhanced carbon source and e-donor supply. 16S rRNA gene sequencing and metagenomic analysis results demonstrated that HM sequestration was accomplished jointly by SRB and HM-resistant bacteria via extracellular precipitation (metal sulfide) and intracellular sequestration, while their contribution depended on the specific coexisting HM species present. This study highlights the critical role of sulfate in the concurrent bioremediation of HM-organohalide composite contamination and provides insights for developing a cost-effective in-situ bioremediation strategy.

Polluticaenibacter yanchengensis gen. nov., sp. nov., A Novel Taxon Within the Family Chitinophagaceae Isolated from Activated Sludge.

A Gram-stain-negative bacterium, designated strain LY-5T, was isolated from an activated sludge sample collected from a river in Yancheng city, Jiangsu province, China. Cells of strain LY-5T, were strictly aerobic, non-motile and rod-shaped. Growth occurred at 15-37°C (optimum, 30°C), at pH 5.0-9.0 (optimum 7.0) and at 0-3% (w/v) NaCl (optimum, 0%). Phylogenetic analysis based on 16S rRNA gene and genome sequences indicated that strain LY-5T formed a distinct phyletic branch within the family Chitinophagaceae, with closest relatives being members of the genera Phnomibacter, Aurantisolimonas, and Chitinophaga, sharing 88.5-90.3% sequence similarity. Moreover, the average amino acid identity (AAI) between strain LY-5T and its closed phylogenetic neighbors was below 70%, indicating it belongs to a novel genus. The predominant cellular fatty acids of LY-5T were iso-C15:0, iso-C15:1 G and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), and the only isoprenoid quinone was menaquinone-7 (MK-7). The major polar lipids identified in strain LY-5T comprised phosphatidylethanolamine, two unidentified aminolipids and five unidentified lipids. The genome size of strain LY-5T was 4.8 Mbp with a G + C content of 38.9%. Based on the evidence presented in this study, strain LY-5T represents a novel species of a new genus in the family Chitinophagaceae, for which the name Polluticaenibacter yanchengensis gen. nov., sp. nov. (Type strain LY-5T = CCTCC AB 2023260T = KCTC 102218T) is proposed.

Comparative Ecotoxicity Assessment of highly bioactive Isomeric monoterpenes carvacrol and thymol on aquatic and edaphic indicators and communities

The growing demand for sustainable natural products to replace harmful synthetic ones requires comprehensive ecotoxicity assessments to ensure their eco-friendly nature. This study explored for the first time the changes in microbial community growth and metabolic profiles from river and natural soil samples exposed to the two structural isomers, thymol (THY) and carvacrol (CARV), utilizing Biolog EcoPlate™ assays and 16S rRNA gene sequencing for taxonomic analysis. In addition, we addressed existing ecotoxicity data gaps for these two compounds by using aquatic (Daphnia magna and Vibrio fischeri) and soil (Eisenia fetida and Allium cepa) indicators. Results show acute toxicity of both CARV and THY on all indicators. V. fischeri (LC50=0.59 mg/L) >D. magna (4.75 mg/L) >A. cepa (6.47 mg/L) for CARV, and V. fischeri (LC50=1.71 mg/L) >A. cepa (4.05 mg/L) >D. magna (8.13 mg/L) for THY. E. fetida showed LC50 = 7.68 mg/Kg for THY and 1.04 for CARV. River and soil microbial communities showed resilience, likely because they contain taxa capable of biodegrading these products. No significant growth inhibition effects were observed up to 100 mg/L, though substrate utilization decreased at higher concentrations, particularly for polymers and amines in soil microorganisms and polymers in aquatic communities. Soil microorganisms were more affected than aquatic ones, with CARV being more toxic than THY (EC50120h = THY 94.13 and CARV 29.79 mg/L in soil microorganisms). These findings suggest that an increase in the consumption of these products and their subsequent ecotoxicity effects from environmental discharge should still be monitored before being ruled out. However, long-term effects are unlikely due to microbial degradation of these natural products, potentially reducing risks to other target species and opening the way for their use as substitutes for commercial antibiotics.

Electrochemical enrichment of a community of weak electricigens and characterisation of three halotolerant electroactive isolates: Micrococcus sp. YH-1, Gordonia sp. RH-1 and Stutzerimonas sp. CH-1

Microbial electroactivity in elevated salt conditions is seldom studied, despite both fundamental and applied relevance. This work reports the enrichment of electroactive microorganisms under saline conditions (2% NaCl), performed on screen-printed carbon electrodes with applied anodic potential. From the weakly electroactive enriched community, three halotolerant electroactive isolates were obtained in pure culture and identified via 16S rRNA gene sequencing. Two isolates were Gram positive (proposed names Micrococcus sp. YH-1 and Gordonia sp. RH-1) and one was Gram negative (proposed name Stutzerimonas sp. CH-1). The isolates were electrochemically characterised on conventional carbon felt electrodes using chronoamperometry. After two days of growth, a current density of 8.29 ± 0.85 μA/cm2 was observed in Micrococcus sp. YH-1, comparable to reported values for the model electroactive microorganism Shewanella oneidensis. Gordonia sp. RH-1 and Stutzerimonas sp. CH-1 appeared to be weak electricigens due to their low current output (2.19 ± 0.46 μA/cm2 and 1.73 ± 0.47 μA/cm2, respectively). The isolates are notable as there are very limited reports of electroactivity in the genera Micrococcus and Gordonia. Cyclic voltammetry revealed prominent redox peaks in Micrococcus sp. YH-1 and Stutzerimonas sp. CH-1. Scanning electron microscopy demonstrated colonisation of the electrode by each isolate, along with thin cellular appendages in Micrococcus sp. YH-1 and Stutzerimonas sp. CH-1. This work extends the catalogue of characterised halotolerant electroactive microbes while also enhancing our understanding of weak electricigens, extracellular electron transfer in Gram positives, and the inherent electroactivity of the natural environment.

No association between the early-life gut microbiota and childhood body mass index and body composition.

The gut microbiota has been implicated in adult obesity, but the causality is still unclear. It has been hypothesized that an obesity-prone gut microbiota can be established in infancy, but only few studies have examined the early-life gut microbiota in relation to obesity in childhood, and no consistent associations have been reported. Here, we examine the association between the early-life gut microbiota and body mass index (BMI) development and body composition throughout childhood. Gut microbiota from stool were collected from 700 children in the Copenhagen Prospective Studies on Asthma in Childhood2010 (COPSAC2010) cohort at ages of 1week, 1month, 1 year, 4 years, and 6 years and analyzed by 16S rRNA gene sequencing. Outcomes included BMI World Health Organization (WHO) Z scores (zBMI), overweight (zBMI>1.04) and obesity (zBMI>1.64) (0-10 years), and adiposity rebound and body composition from dual-energy X-ray absorptiometry at 6 years. The early-life gut microbiota diversity, overall composition, and individual taxon abundances in unsupervised and supervised models were not consistently associated with either current or later BMI Z scores, overweight, obesity, adiposity rebound, or body composition in childhood. In a deeply characterized longitudinal birth cohort, we did not observe any consistent associations between the early-life gut microbiota and BMI or risk of obesity in later childhood. While this does not conclusively rule out a relationship, it suggests that if such associations exist, they may be more complex and potentially influenced by factors emerging later in life, including lifestyle changes. COPSAC is funded by private and public research funds (all listed on www.copsac.com).

Pseudomonas huanghezhanensis sp. nov. and Pseudomonas fjordensis sp. nov., two novel species isolated from seawater of Arctic Kongsfjorden in the Ny-Ålesund area, Svalbard.

Two Gram-stain-negative bacterial strains, BSw22131T and DMSP-1T, were isolated from seawater of Arctic Kongsfjorden in the Ny-Ålesund area, Svalbard. Strains BSw22131T and DMSP-1T were within the genus Pseudomonas and presented 98.1 and 99.8% 16S rRNA gene sequence similarity to Pseudomonas piscicola P50T and Pseudomonas paracarnis V5/DAB/2/5T, respectively. However, the results of multilocus sequence analysis (concatenated 16S rRNA, gyrB, rpoB and rpoD gene sequences) and whole-genome sequence analysis indicated that BSw22131T and DMSP-1T were most closely related to Pseudomonas petrae P2653T and Pseudomonas lactis DSM 29167T, respectively. The genome G+C contents of strains BSw22131T and DMSP-1T were 58.2 and 60.0%, respectively. The two isolates presented digital DNA-DNA hybridization and average nucleotide identity values of less than 70 and 96%, respectively, with type strain genomes of their close relatives. BSw22131T contained summed feature 3 (C16 : 1 ω6c/C16 : 1 ω7c), C16 : 0 and summed feature 8 (C18 : 1 ω6c/C18 : 1 ω7c) as the major cellular fatty acids, whereas DMSP-1T contained an additional C17 : 0-cyclo as the major fatty acid. The major quinones of both strains were Q-9 (57.5-77.2%) and Q-8 (22.8-37.2%), and their major cellular polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. BSw22131T could be differentiated from its related species by positive ornithine decarboxylase, esterase (C8) and l-rhamnose and d-fructose-6-PO4 utilization, whereas DMSP-1T could be differentiated by positive acid phosphatase but negative p-hydroxyphenylacetic acid utilization. The two strains were negative for the reduction of nitrates but contained genes related to dimethylsulfoniopropionate catabolism. The results of the polyphasic taxonomy analysis revealed that strains BSw22131T and DMSP-1T represent two novel species of the genus Pseudomonas, for which the names Pseudomonas huanghezhanensis sp. nov. (type strain BSw22131T = CCTCC AB 2019135T = JCM 36635T = DSM 117336T) and Pseudomonas fjordensis sp. nov. (type strain DMSP-1T = CCTCC AB 2019124T = JCM 36636T = DSM 117337T) are proposed.

Mercury-tolerant metalophiles: A bio tool for remediation of mercury (Hg) affected Environs

Due to their low solubility, carcinogenic and mutagenic effects, heavy metals are of critical concern to humans and the environment. Mercury (Hg) is one of the rarest and least concentrated elements in the world, but it is ranked third in terms of toxicity. The present investigation evaluates the response and tolerance limit of bacteria isolated from the saffron soil of the Kashmir Valley to various mercury concentrations (50 ppm to 400 ppm). The minimum inhibitory concentration (MIC) was calculated after 24hours by measuring the optical density. Isolated bacteria have been characterized using two efficient methods, i.e. 16S rRNA gene sequence and MALDI-TOF MS. A total of 72 isolates were investigated for the mercury tolerance and among them, only 8 isolates (Staphylococcus equorum, Leclercia adecarboxylata, Staphylococcus warneri, Klebsiella pneumoniae, Serratia plymuthica, Bacillus megaterium, Pseudomonas putida and Bacillus safensis) were found tolerant to mercury with varying MIC. In comparison, Pseudomonas putida has been found to have the maximum resistance (400 ppm) for mercury and S. plymuthica and L. adecarboxylate had the lowest resistance level (200 ppm). The sequestration potential results revealed that P. putida executed the highest sequestration potential (73%) while K. pneumoniae showed the lowest sequestration potential (51.63%). Thus mercury tolerance capacity of these species may be utilised for detoxification and removal of mercury from polluted sites, providing an eco-friendly technique for bioremediation of mercury-affected environments.

Anthropogenic activities affect the diverse autotrophic communities of coastal sediments

Coastal sediments are a critical domain for carbon sequestration and are profoundly impacted by human activities. Therefore, it is essential to understand the structure and components of benthic autotrophs that play a crucial role in carbon sequestration processes, as well as the influence of anthropogenic activities on their communities. This study utilized an urban estuary, an industrial sea bay, a maricultural sea region, and two mangrove coastlines within the coastal areas of Guangdong Province, China. The micro-benthos in these environments, including prokaryotes and eukaryotes, were identified through high-throughput sequencing of 16S rRNA and 18S rRNA genes. The findings show that the autotrophic composition was altered by the interactions of anthropogenic heavy metals (Cd and Zn) and micro-eukaryotes (protazoa, metazoa, and parasitic organisms). Industrial pollution reduced the abundance of both prokaryotic and eukaryotic autotrophs. Mangroves induced a substantial transformation in the sediment eukaryotic and prokaryotic composition, increasing the proportion of autotrophs, notably sulfur-oxidizing and iron-oxidizing bacteria and microalgae. This alteration suggests an increase in specific sulfur and iron cycling with simultaneous carbon sequestration within mangrove sediments. These results indicate that anthropogenic activities affect sediment carbon sequestration by altering autotrophic assemblages along coastlines, thereby inducing consequential shifts in overall elemental cycling processes.

Undernutrition disrupts jejunal and ileal microbiota and epithelial tissue homeostasis in a pregnant sheep model

Nutrition consistently affects microbe-host interactions in the gastrointestinal tract. This study aimed to unravel how undernutrition reshapes the microbial composition and the homeostasis of epithelium in the jejunum and ileum. Sixteen late-gestation Hu-sheep were randomly assigned to the control group (n = 8, 100% ad libitum feeding levels) or the undernutrition group (n = 8, which received 30% ad libitum feeding levels). After 15-d treatment, all ewes were slaughtered, and jejunal and ileal digesta and epithelium samples were collected for 16S rRNA gene sequencing and transcriptome sequencing, respectively. Results indicated that undernutrition decreased the jejunal and ileal tissue weights (P = 0.005 and P = 0.022) and the levels of volatile fatty acids (P = 0.019 and P = 0.007) and microbial protein levels (P = 0.019 and P = 0.031) in jejunal and ileal digesta. The relative abundance of acetate producing microbiota, including Clostridia UCG-014 norank, Ruminococcus, [Ruminococcus] gauvreauii, and Lachnospiraceae_Blautia, were significantly reduced (P < 0.05) in the jejunum and ileum. Undernutrition up-regulated (P < 0.05) the expression of genes involved in amino acid synthesis and fatty acid oxidation, but down-regulated (P < 0.05) the expression of genes associated with amino acid degradation, fatty acid synthesis, and extracellular structures in jejunal and ileal epithelium. In the jejunal epithelium, genes associated with extracellular matrix-receptor interactions, cell growth, and immune response were down-regulated (P < 0.05) upon undernutrition. Taken together, undernutrition changed the microbial community in the jejunum and ileum, which altered the fermentation mode and the production of volatile fatty acids and microbial protein. These affected the energy and protein system in the epithelium and reprogrammed substance metabolism and extracellular structures, which probably further influenced cell growth and immune response. These insights provide a foundation for completely clarifying the crosstalk between small intestinal microbiota and the host.

16S rRNA gene sequencing for bacterial identification and infectious disease diagnosis

16S rRNA gene sequencing for bacterial identification and infectious disease diagnosis

Features of the oropharyngeal microbiota of healthy children and those with acute respiratory infections. A prospective single-center randomized study

Background. The oropharyngeal microbiota is involved in the development of colonization resistance, affecting viral adhesion and metabolism. Any deviations in the stability of the environmental microbial niches of the oropharynx alter the local immune response and can trigger severe chronic somatic disorders. Aim. To compare the composition of the microbiota of children during an episode of respiratory infection and healthy volunteers examined in different periods of convalescence. The obtained data can be used to predict the course of the disease, analyze the risk of complications, and assess the frequency of the diseases in the future. Materials and methods. From 20.01.2022 to 23.12.2022, an open-label prospective single-center randomized comparative study was conducted, which included 120 children aged 5-10 who presented with symptoms of acute respiratory infection. The control group consisted of 15 asymptomatic children examined at different periods of convalescence. The study compared changes in the oral microbiota composition of patients and healthy children. The evaluation was performed using complete 16S rRNA gene sequencing on the Oxford Nanopore platform. Taxonometric analysis at the species and genera level was performed using the Knomics-Biota platform. The R programming language was used for statistical processing. In addition, the parameters of α- and β-diversity of the microbiota were assessed. The Chao1 and Shannon indices were calculated to assess α-diversity (diversity within one community). The Chao1 index is based on the number of species found in the sample and also rare species that are found only 1 or 2 times, thus preventing underestimation of diversity. The Shannon index includes both the number of species and their uniformity in the community. A change in the index indicates the dominance of one or more species. The following indicators were used to assess the β-diversity describing the differences between two microbiota samples at the species and genera level: Bray–Curtis dissimilarity (characterizing the ratios of common and different microorganisms between the samples) and the Aitchison distance (reflecting the differences in the proportions of microorganisms). In addition, the balance between the two groups of microorganisms was evaluated using the NearesBalance method, which characterizes the differences between the microbiota of the main and control groups. Results. The results of our study show that during respiratory infection, the state of the oropharyngeal microbiota is characterized by an increase in α-diversity, which is associated with an increase in the proportion of species of Streptococcus salivarius, Streptococcus pneumoniae, Streptococcus pseudopneumoniae, Streptococcus pyogenes, Streptococcus thermophilus and A12 versus the proportion of Streptococcus mitis, Streptococcus oralis, Streptococcus gwangjuense, Streptococcus sanguinis, Streptococcus gordonii and FDAARGOS_192. In the control group, the oropharyngeal microbiota was divided into two clusters. The dominance of streptococci was observed in the first group (control), while the second group had a more uniform representation. Analysis of the changes in microbial communities in the main and control groups can show the stages of oral microbiota recovery after an acute respiratory infection episode. Conclusion. More research is needed in the field of the oropharyngeal microbiome on a larger sample of healthy children, to standardize the communities of the oropharyngeal microbiota, to study the interaction within communities and with the body as a whole. Analysis of the impact of the microbiota on the frequency and course of respiratory infections and the rate of complications opens up new prospects in treating, rehabilitating, and preventing diseases. These research areas can contribute to improving children's health and quality of life and introducing new approaches into clinical practice.

Prospects of iron solubilizing Bacillus species for improving growth and iron in maize (Zea mays L.) under axenic conditions

Iron (Fe) deficiency in calcareous soils is a significant agricultural challenge, affecting crop productivity and nutritional quality. This study aimed to isolate, characterize, and evaluate Fe solubilizing rhizobacterial isolates from maize rhizosphere in calcareous soils as potential biofertilizers. Forty bacterial isolates coded as SG1, SG2, …, SG40 were isolated and screened for siderophore production, with ten showing significant Fe solubilizing capabilities. These isolates were further assessed for phosphate solubilization and exopolysaccharides production. The selected bacterial isolates were also screened under axenic conditions for their ability to improve maize growth. The isolates SG8, SG13, SG24, SG30 and SG33 significantly enhanced growth parameters of maize. Notably, SG30 showed highest increment in shoot length (58%), root length (54%), root fresh and dry biomass (67% and 76%), SPAD value (67%), relative water contents (69%), root surface area (61%), and Fe concentration in shoots (79%) as compared to control. The biochemical characterization of these strains showed that all these strains have capability to solubilize insoluble phosphorus, produce indole-3-acetic acid (IAA), and ammonia with catalase, urease and protease activity. Molecular identification through 16s rRNA gene sequencing confirmed high similarity (99.7–100%) of the selected isolates to various Bacillus species, including B. pyramidoids, B. firmicutes, and B. cereus. The study provides a strong base for developing eco-friendly, cost-effective biofertilizers to address Fe deficiency in crops and promote sustainable agriculture.

Microbiome and network analysis reveal potential mechanisms underlying Carassius auratus diseases: The interactions between critical environmental and microbial factors

Despite the rapid development of research on aquatic environment microbiota, limited attention has been paid to exploring the complex interactions between microbial communities and aquatic environments. Particularly, the mechanisms underlying fish diseases based on such dynamic interactions remain unknown. This study aimed to address the gap by conducting microbiome and co-occurrence network analyses on the typical freshwater aquaculture systems. High-throughput 16S rRNA gene sequencing results revealed significant differences in the microbiota between the disease and healthy groups. Notably, disease mortality varied consistently with the gradient of relative abundance of Proteobacteria (intestine, R2 = 0.46, p < 0.05) and Cyanobacteria (gill, R2 = 0.67, p < 0.01), indicating their potential use as diagnostic criteria. Furthermore, the elevated hepatosomatic index, NO3-N, COD and TC (sediment) were directly correlated with diseases (r > 0.54, p < 0.01). Mean concentrations of NO3-N, COD and TC were elevated by 78.87%, 25.63% and 44.2%, respectively, in ponds where diseases occurred. Quantitative analysis (qPCR) revealed that Aeromonas sobria infected hosts through a potential pathway of “sediment (4.4 × 105 copy number/g)-water (1.1 × 103 copy number/mL)-intestine (1.2 × 106 copy number/g)”. Similarly, the potential route for Aeromonas veronii was sediment (4.9 × 106 copy number/g) to gill (5.1 × 105 copy number/g). Additionally, the complexity of microbial networks in the intestine, water, and sediment was significantly lower in the disease group, although no similar phenomenon was observed in the gill microbial network. In summary, these findings reveal that elevated concentrations of crucial environmental factors disrupt the linkages within microbiota, fostering the growth of opportunistic bacteria capable of colonizing fish gut or gills. This offers new insights into potential mechanisms by which environmental factors cause disease in fish.

Peribacillus aracenensis sp.nov., a plant growth promoting bacteria for agriculture in water-scarce conditions isolated from Pinus pinaster rhizosphere

A gram-positive, nonpathogenic, central endospore-forming, flagellated strain, was successfully isolated from the rhizosphere of Pinus pinaster in Aracena (Spain). Its optimal growth conditions are 28 °C, pH 6, and 0 % salinity. It is able to assimilate glucose, L-fucose, L-arabinose, b-metil-D-xylose and shows high catabolic capacity. The major fatty acids (>79.20 % of the total fatty acids) are anteiso C15:0 > iso C15:0 > C14:0.A phylogenetic analysis based on the 16S rRNA gene sequence revealed similarity to P. frigoritolerans DSM8801T (99.9 %), P. castrilensis CECT30509T (99.8 %), and P. simplex DSM1321T (99.6 %). Comparison of whole genomes revealed that strain BBB004T is more similar to P. simplex DSM1321T. According to ANI (93.54 %), AAI (94 %), dDDH (60.6), %G + C (0.12), TETRA (0.99822) and intergenomic distance (0.2835) values, therefore this species is different to the closest. A total of 133 genes unique to Peribacillus BBB004T were identified.Supported by these analyses, strain BBB004T (=LMG32742T = CCUG76477T) was proposed as the type strain for a new species, named “Peribacillus aracenensis sp. nov.”

Cryptic diversity patterns of subterranean estuaries.

Subterranean estuaries are coastal ecosystems characterized by vertically stratified groundwater. The biota within these ecosystems is relatively understudied due to the inherent difficulty of accessing such extreme environments. The fauna inhabiting these ecosystems is considered vulnerable to extinction, and the presence of cryptic species has major implications for research and conservation efforts. Most species lack molecular data; however, the evaluation of genetic data for some taxa has revealed that undocumented species are common. This study employs molecular species delimitation methods and DNA barcoding through the analysis of publicly and newly generated sequences, including individuals from type localities and non-crustacean phyla; the latter are typically overlooked in biodiversity assessments of subterranean estuaries. We analysed 376 cytochrome c oxidase subunit I (COI) gene sequences and 154 16S rRNA gene sequences. The COI sequences represented 32% of previously described species and 50% of stygobiont species from the Yucatan Peninsula and Cozumel Island, while sequences of the 16S rRNA represented 14% of described species and 22% of stygobionts. Our results revealed cryptic genetic lineages and taxonomic misidentification of species. As several species from these ecosystems are recognized as endangered, the use of molecular approaches will improve biodiversity estimates and highlight overlooked cryptic lineages in need of evaluation of conservation status.

Antibiotic Resistance Genes in Plague Ecosystems: Threatening the Emergence of Resistant Plague

The study aimed to investigate the prevalence of antibiotic resistance genes (ARGs) within the ecosystem of natural plague foci, assessing their potential impact on the efficacy of plague treatments. Employing 16S rRNA gene sequencing and high-throughput quantitative PCR, microbial communities and ARGs were detected, with subsequent analysis of interactions among ARGs, mobile genetic elements (MGEs), environmental factors, and microbial species. Tetracycline resistance genes were found to be dominant, with multidrug and tetracycline resistance ARGs primarily associated with marmots and ecological soil, while pikas predominantly harbored β-lactam resistance ARGs. High detection rates were observed for resistance genes rpsl and sul1, which are relevant to streptomycin and sulfonamides, antibiotics commonly used in plague treatment. The total dissolved solids (TDS) in soil significantly promoted the presence of tetR-02, and Ni was found to inhibit vanHB. The tnpA-03 MGE was identified as a significant contributor to the dissemination of the aadE gene. The high prevalence of ARGs, particularly rpsl and sul1, poses a potential risk to the efficacy of main antibiotic treatments for plague. The study suggests that environmental microbiomes may be the greatest risk factor for the emergence of drug-resistant Yersinia pestis, given the low misuse of antibiotics in animals within natural plague foci. Monitoring the risk of drug-resistant strain emergence and preparing alternative antibiotic or combination therapy strategies based on ARG pollution levels in plague-affected areas is deemed necessary.