Microbial Ecology Research Articles

Comparative analysis of size-fractional eukaryotic microbes in subtropical riverine systems inferred from 18S rRNA gene V4 and V9 regions

Eukaryotic microbes play key ecological roles in riverine ecosystems. Amplicon sequencing has greatly facilitated the identification and characterization of eukaryotic microbial communities. Currently, 18S rRNA gene V4 and V9 hypervariable regions are widely used for sequencing eukaryotic microbes. Identifying optimal regions for the profiling of size-fractional eukaryotic microbial communities is critical for microbial ecological studies. In this study, we spanned three rivers with typical natural-human influenced transition gradients to evaluate the performance of the 18S rRNA gene V4 and V9 hypervariable regions for sequencing size-fractional eukaryotic microbes (>180 μm, 20-180 μm, 5-20 μm, 3-5 μm, 0.8-3 μm). Our comparative analysis revealed that amplicon results depend on the specific species and microbial size. The V9 region was most effective for detecting a broad taxonomic range of species. The V4 region was superior to the V9 region for the identification of microbes in the minor 3 μm and at the family and genus levels, especially for specific microbial groups, such as Labyrinthulomycetes. However, the V9 region was more effective for studies of diverse eukaryotic groups, including Archamoebae, Heterolobosea, and Microsporidia, and various algae, such as Haptophyta, Florideophycidae, and Bangiales. Our results highlight the importance of accounting for potential misclassifications when employing both V4 and V9 regions for the identification of microbial sequences. The use of optimal regions for amplification could enhance the utility of amplicon sequencing in environmental studies. The insights gained from this work will aid future studies that employ amplicon-based identification approaches for the characterization of eukaryotic microbial communities and contribute to our understanding of microbial ecology within aquatic systems.

Molecular Identification of Soil Bacteria

The molecular identity of soil micro-organism is pivotal for expertise soil ecosystems that are important for agricultural productiveness, environmental sustainability, and biogeochemical cycles. This take a look at employs superior molecular techniques, inclusive of high-throughput sequencing and bioinformatics gear, to identify and classify micro-organism in various soil samples. Our comprehensive evaluation discovered a wealthy variety of bacterial species, which include both known and novel taxa, illustrating the complexity of soil micro biomes. These findings emphasize the vast function of soil micro-organism in nutrient cycling and plant health, supplying treasured insights for enhancing soil management practices and agricultural results. Additionally, the observe highlights the efficacy of molecular strategies in microbial ecology, suggesting potential future research instructions to take advantage of soil bacteria for environmental and biotechnological packages. This paintings underscores the crucial importance of molecular identification in unveiling the intricacies of soil bacterial groups and their useful roles within the ecosystem.

Gut metagenome in Insects and Its Potential in Agricultural and Industrial Applications

Metagenomics has greatly improved our understanding of microbial ecology by revealing the metagenomes of uncultured bacteria, including those associated with insects. These bacteria play important roles in insect defence, reproduction, and food absorption, which influences pest management tactics. Recent next generation sequencing advances in low-cost nucleotide and user-friendly bioinformatics have improved our ability to investigate microbial diversity in economically significant pests. Meta-omics technologies are now essential for defining microbial ecosystems, bio-surveillance, food safety, and commercial advances. Genome studies on insects and plants provide insights into pest control by discovering resistance genotypes and understanding stress-induced genomic changes. This review is focused on how herbivore specialists, such as the diamondback Plutella xylostella, adapt to host plants using gut microbiota. Metagenomic sequencing has discovered important bacteria-Enterobacter cloacae, Enterobacter asburiae, and Carnobacterium maltaromaticum- that help detoxify plant defences, breakdown cell walls, and synthesise amino acids. These findings indicate novel pest management tactics that target gut microbiota interactions. The paper continues with an overview of metagenomic DNA extraction, library creation and screening methods, especially on their applications in biotechnology and bioprocessing.

Breed-Driven Microbiome Heterogeneity Regulates Intestinal Stem Cell Proliferation via Lactobacillus-Lactate-GPR81 Signaling.

Genetically lean and obese individuals have distinct intestinal microbiota and function. However, the underlying mechanisms of the microbiome heterogeneity and its regulation on epithelial function such as intestinal stem cell (ISC) fate remain unclear. Employing pigs of genetically distinct breeds (obese Meishan and lean Yorkshire), this study reveals transcriptome-wide variations in microbial ecology of the jejunum, characterized by enrichment of active Lactobacillus species, notably the predominant Lactobacillus amylovorus (L. amylovorus), and lactate metabolism network in obese breeds. The L. amylovorus-dominant heterogeneity is paralleled with epithelial functionality difference as reflected by highly expressed GPR81, more proliferative ISCs and activated Wnt/β-catenin signaling. Experiments using in-house developed porcine jejunal organoids prove that live L. amylovorus and its metabolite lactate promote intestinal organoid growth. Mechanistically, L. amylovorus and lactate activate Wnt/β-catenin signaling in a GPR81-dependent manner to promote ISC-mediated epithelial proliferation. However, heat-killed L. amylovorus fail to cause these changes. These findings uncover a previously underrepresented role of L. amylovorus in regulating jejunal stem cells via Lactobacillus-lactate-GPR81 axis, a key mechanism bridging breed-driven intestinal microbiome heterogeneity with ISC fate. Thus, results from this study provide new insights into the role of gut microbiome and stem cell interactions in maintaining intestinal homeostasis.

Long-Term Exposure to Polystyrene Microspheres and High-Fat Diet-Induced Obesity in Mice: Evaluating a Role for Microbiota Dysbiosis.

Microplastics (MPs) have become a global environmental problem, emerging as contaminants with potentially alarming consequences. However, long-term exposure to polystyrene microspheres (PS-MS) and its effects on diet-induced obesity are not yet fully understood. We aimed to investigate the effect of PS-MS exposure on high-fat diet (HFD)-induced obesity and underlying mechanisms. In the present study, C57BL/6J mice were fed a normal diet (ND) or a HFD in the absence or presence of PS-MS via oral administration for 8 wk. Antibiotic depletion of the microbiota and fecal microbiota transplantation (FMT) were performed to assess the influence of PS-MS on intestinal microbial ecology. We performed 16S rRNA sequencing to dissect microbial discrepancies and investigated the dysbiosis-associated intestinal integrity and inflammation in serum. Compared with HFD mice, mice fed the HFD with PS-MS exhibited higher body weight, liver weight, metabolic dysfunction-associated steatotic liver disease (MASLD) activity scores, and mass of white adipose tissue, as well as higher blood glucose and serum lipid concentrations. Furthermore, 16S rRNA sequencing of the fecal microbiota revealed that mice fed the HFD with PS-MS had greater -diversity and greater relative abundances of Lachnospiraceae, Oscillospiraceae, Bacteroidaceae, Akkermansiaceae, Marinifilaceae, Deferribacteres, and Desulfovibrio, but lower relative abundances of Atopobiaceae, Bifidobacterium, and Parabacteroides. Mice fed the HFD with PS-MS exhibited lower expression of MUC2 mucin and higher levels of lipopolysaccharide and inflammatory cytokines [tumor necrosis factor- (TNF-), interleukin-6 (IL-6), IL-1, and IL-17A] in serum. Correlation analyses revealed that differences in the microbial flora of mice exposed to PS-MS were associated with obesity. Interestingly, microbiota-depleted mice did not show the same PS-MS-associated differences in Muc2 and Tjp1 expression in the distal colon, expression of inflammatory cytokines in serum, or obesity outcomes between HFD and HFD + PS-MS. Importantly, transplantation of feces from HFD + PS-MS mice to microbiota-depleted HFD-fed mice resulted in a lower expression of mucus proteins, higher expression of inflammatory cytokines, and obesity outcomes, similar to the findings in HFD + PS-MS mice. Our findings provide a new gut microbiota-driven mechanism for PS-MS-induced obesity in HFD-fed mice, suggesting the need to reevaluate the adverse health effects of MPs commonly found in daily life, particularly in susceptible populations. https://doi.org/10.1289/EHP13913.

Sample Preservation Solution Increases Nucleic Acid Yield and Environmental RNA Quality in Sediments Across an Estuarine Salinity Gradient

ABSTRACTEnvironmental nucleic acid‐based assessments are powerful tools for understanding microbial ecology, and environmental degradation in aquatic environments. This approach is particularly useful in guiding restoration in estuaries, some of the most degraded ecosystems in the world. The recent popularity of this approach has been accompanied by a parallel increase in the diversity of applied methods. A range of best practice methods exist across the field that can be employed and are selected based on environmental considerations such as physicochemical gradients, maximizing yield, and quality of nucleic acids sampled across sites within a study area. A consistent approach to intra‐study nucleic acid sampling also ensures accurate comparison between those sites. This study evaluates environmental nucleic acid (eNA) sampling methods across salinity gradients in aquatic ecosystems, focusing on the impact of preservation techniques on environmental DNA (eDNA) yield and environmental RNA (eRNA) yield and quality. Fieldwork was conducted at three sites within the Coorong estuary system in South Australia, representing low salinity, marine, and hypersaline conditions. Snap freezing and LifeGuard preservation solution treatments were applied in situ to compare their effects on nucleic acid yields and eRNA integrity. Snap freezing enhanced eDNA yield in low salinity sediments but negatively impacted eRNA integrity in marine and hypersaline conditions. Conversely, treatment with preservation solution consistently improved both eDNA and eRNA recovery across all salinity levels, which makes this approach a good candidate for preserving eNA molecules across environmental gradients. The study underscores the necessity of tailoring sample preservation methods to specific environmental conditions for accurate eNA‐based microbial community assessments in coastal ecosystems. These findings contribute to the development of robust eNA sampling protocols for benthic communities under varying salinity conditions.

Is everything everywhere? A hands-on activity to engage undergraduates with key concepts in quantitative microbial biogeography.

The ubiquity and ease with which microbial cells disperse over space is a key concept in microbiology, especially in microbial ecology. The phenomenon prompted Baas Becking's famous "everything is everywhere" statement that now acts as the null hypothesis in studies that test the dispersal limitation of microbial taxa. Despite covering the content in lectures, exam performance indicated that the concepts of dispersal and biogeography challenged undergraduate students in an upper-level Microbial Ecology course. Therefore, we iteratively designed a hands-on classroom activity to supplement the lecture content and reinforce fundamental microbial dispersal and biogeography concepts while also building quantitative reasoning and teamwork skills. In a class period soon after the lecture, the students formed three-to-five-person teams to engage in the activity, which included a hands-on dispersal simulation and worksheet to guide discussion. The simulation involved stepwise neutral immigration or emigration and then environmental selection on a random community of microbial taxa represented by craft poms. The students recorded the results at each step as microbial community data. A field guide was provided to identify the taxonomy based on the pom phenotype and a reference to each taxon's preferred environmental niches. The worksheet guided a reflection of student observations during the simulation. It also sharpened quantitative thinking by prompting the students to summarize and visualize their and other teams' microbial community data and then to compare the observed community distributions to the idealized expectation given only selection without dispersal. We found that the activity improved student performance on exam questions and general student satisfaction and comfort with the biogeography concepts. Activity instructions and a list of needed materials are included for instructors to reproduce for their classrooms.

A Microbial Phenomics Approach to Determine Metabolic Signatures to Enhance Seabream Sparus aurata Traceability, Differentiating between Wild-Caught and Farmed.

The need for efficient and simplified techniques for seafood traceability is growing. This study proposes the Biolog EcoPlate assay as an innovative method for assessing wild and farmed Sparus aurata traceability, offering advantages over other molecular techniques in terms of technical simplicity. The Biolog EcoPlate assay, known for its high-throughput capabilities in microbial ecology, was utilized to evaluate the functional diversity of microbial communities from various organs of S. aurata (seabream) from the Mediterranean area. Samples were taken from the anterior and posterior gut, cloaca swabs and gills to distinguish between farmed and wild-caught individuals. The analysis focused on color development in OmniLog Units for specific carbon sources at 48 h. Gills provided the most accurate clusterization of sample origin. The assay monitored the development of color for carbon sources such as α-cyclodextrin, D-cellobiose, glycogen, α-D-lactose, L-threonine and L-phenylalanine. A mock experiment using principal component analysis (PCA) successfully identified the origin of a blind sample. Shannon and Simpson indexes were used to statistically assess the diversity, reflecting the clusterization of different organ samples; Conclusions: The Biolog EcoPlate assay proves to be a quick, cost-effective method for discriminate S. aurata traceability (wild vs. farmed), demonstrating reliable reproducibility and effective differentiation between farmed and wild-caught seabream.

A Mathematical Model of Diel Activity and Long Time Survival in Phototrophic Mixed-Species Subaerial Biofilms

Subaerial biofilms (SAB) are intricate microbial communities living on terrestrial surfaces, of interest in a variety of contexts including cultural heritage preservation, microbial ecology, biogeochemical cycling, and biotechnology. Here we propose a mathematical model aimed at better understanding the interplay between cyanobacteria and heterotrophic bacteria, common microbial SAB constituents, and their mutual dependence on local environmental conditions. SABs are modeled as thin mixed biofilm-liquid water layers sitting on stone. A system of ordinary differential equations regulates the dynamics of key SAB components: cyanobacteria, heterotrophs, polysaccharides and decayed biomass, as well as cellular levels of organic carbon, nitrogen and energy. These components are interconnected through a network of energetically dominant metabolic pathways, modeled with limitation terms reflecting the impact of biotic and abiotic factors. Daily cylces of temperature, humidity, and light intensity are considered as input model variables that regulate microbial activity by influencing water availability and metabolic kinetics. Relevant physico-chemical processes, including pH regulation, further contribute to a description of the SAB ecology. Numerical simulations explore the dynamics of SABs in a real-world context, revealing distinct daily activity periods shaped by water activity and light availability, as well as longer time scale survivability conditions. Results also suggest that heterotrophs could play a substantial role in decomposing non-volatile carbon compounds and regulating pH, thus influencing the overall composition and stability of the biofilm.

Metagenomic insights into bacterial dynamics and niche partitioning in response to varying oxygen gradients in the Arabian Sea oxygen minimum zone (OMZ)

This study investigates the impact of oxygen gradients on bacterial diversity and community composition within the Arabian Sea oxygen minimum zone (OMZ), providing insight into their ecology in this unique environment. Vertical profiling of physicochemical parameters and bacterial community composition was conducted at two stations within the OMZ using Illumina sequencing of the V3-V4 regions of the 16S rRNA gene. The oxygen levels in the water column of the study locations varied from oxic to hypoxic and Station-I exhibited intense anoxic conditions. Bacterial diversity analyses revealed a rich assemblage of 24 bacterial phyla dominated by Proteobacteria, with significant presence of Actinobacteria and Planctomycetes. SIMPER (Similarity percentage) analysis highlights taxa such as Synechococcophycideae, Flavobacteriia in oxic water and Phycisphaerae as significant contributors to community dissimilarity in oxygen-deficient water, emphasizing their ecological roles in nutrient cycling and the impact of oxygen availability on marine biodiversity. Canonical Correspondence Analysis (CCA) further elucidates the relationships between bacterial communities and environmental variables, with oxygen, temperature, and nutrient concentrations playing crucial roles in shaping bacterial community distributions. This research enhances our understanding of microbial ecology in the Arabian Sea OMZ by revealing how microbial communities respond to changing oxygen conditions, providing insights into the ecological dynamics of the region. Furthermore, these findings underscore the need for continued research to predict and mitigate the effects of environmental changes on marine ecosystems, particularly in the context of expanding OMZs and global biogeochemical cycles.

A pangenomic atlas reveals eco-evolutionary dynamics that shape type VI secretion systems in plant-pathogenic Ralstonia.

Soilborne Ralstonia solanacearum species complex (RSSC) pathogens disrupt microbial communities as they invade roots and fatally wilt plants. RSSC pathogens secrete antimicrobial toxins using a type VI secretion system (T6SS). To investigate how evolution and ecology have shaped the T6SS of these bacterial pathogens, we analyzed the T6SS gene content and architecture across the RSSC and their evolutionary relatives. Our analysis reveals that two ecologically similar Burkholderiaceae taxa, xylem-pathogenic RSSC and Paracidovorax, have convergently evolved to wield large arsenals of T6SS toxins. To understand the mechanisms underlying genomic enrichment of T6SS toxins, we compiled an atlas of 1,066 auxiliary T6SS toxin clusters ("aux" clusters) across 99 high-quality RSSC genomes. We classified 25 types of aux clusters with toxins that predominantly target lipids, nucleic acids, or unknown cellular substrates. The aux clusters were located in diverse genetic neighborhoods and had complex phylogenetic distributions, suggesting frequent horizontal gene flow. Phages and other mobile genetic elements account for most of the aux cluster acquisition on the chromosome but very little on the megaplasmid. Nevertheless, RSSC genomes were more enriched in aux clusters on the megaplasmid. Although the single, ancestral T6SS was broadly conserved in the RSSC, the T6SS has been convergently lost in atypical, non-soilborne lineages. Overall, our data suggest dynamic interplay between the lifestyle of RSSC lineages and the evolution of T6SSes with robust arsenals of toxins. This pangenomic atlas poises the RSSC as an emerging, tractable model to understand the role of the T6SS in shaping pathogen populations.IMPORTANCEWe explored the eco-evolutionary dynamics that shape the inter-microbial warfare mechanisms of a globally significant plant pathogen, the Ralstonia solanacearum species complex. We discovered that most Ralstonia wilt pathogens have evolved extensive and diverse repertoires of type VI secretion system-associated antimicrobial toxins. These expansive toxin arsenals potentially enhance the ability of Ralstonia pathogens to invade plant microbiomes, enabling them to rapidly colonize and kill their host plants. We devised a classification system to categorize the Ralstonia toxins. Interestingly, many of the toxin gene clusters are encoded on mobile genetic elements, including prophages, which may be mutualistic symbionts that enhance the inter-microbial competitiveness of Ralstonia wilt pathogens. Moreover, our findings suggest that the convergent loss of this multi-gene trait contributes to genome reduction in two vector-transmitted lineages of Ralstonia pathogens. Our findings demonstrate that the interplay between microbial ecology and pathogen lifestyle shapes the evolution of a genetically complex antimicrobial weapon.

Quantifying the functional genes of C, N, P, and S cycling in a deep lake: Depth patterns and drivers

Microbial functional composition is important for biogeochemical cycles, and is usually constrained by taxonomic species pool and natural environmental gradients. However, the distribution of functional genes in deep lakes and the driving factors remains elusive. Here, we quantified the abundance of 71 functional genes relevant to carbon degradation, carbon fixation, methane metabolism, nitrogen cycling, phosphorus cycling, and sulfur cycling in 38 sediments along the water depth ranging from 0 to 90 m in Lugu Lake, China, using Quantitative Microbial Ecology Chip (QMEC) and then explored their water-depth diversity pattern and abiotic and biotic drivers. Functional gene diversity showed a hump-shaped pattern along the depth and peaked at around 50 m which is the low boundary of thermocline layer. There were specific environmental preferences among functional gene subgroups such as nitrogen and sulfur cycling genes preferring to deeper and shallow waters, respectively. The dissimilarity of total functional genes increased with water depth distance indicating a distance-decay relationship. There was a congruence between functional and taxonomic composition by showing the positive correlations between the compositions of functional genes and bacteria or archaea. This phenomenon is consistently observed for the six functional gene subgroups, and the congruence strength was highest for nitrogen cycling while lowest for sulfur cycling. Compared to abiotic factors, biotic factors were more relevant to the functional gene diversity and composition. Biotic factors explained 28.5 % of the variance of functional gene diversity, while water depth and other environmental factors such as water total phosphorus and sediment carbon explained 4.2 % and 3.8 %, respectively. For functional gene composition, biotic factors accounted for 25.2 % of the variance, whereas water depth and other environmental factors contributed to 0.7 % and 4.4 %, respectively. Among all explanatory variables for function genes, bacterial composition had the highest contribution, which was further supported by showing its direct effects of 1.45 and 0.86 on functional diversity and composition, respectively. This study for the first time quantified the microbial functional genes along water depth in deep lakes and provided a more comprehensive understanding of the functional dynamics in microbial communities within aquatic ecosystems.

Metagenomic Profiling of Bacterial Communities and Functional Genes in Penaeus monodon

Penaeus monodon is one of the world’s most important aquaculture species, with its host-associated microbial community playing a crucial role in its growth, metabolism, immune response, and adaptation. In our study, we utilized Illumina high-throughput sequencing to investigate the composition, structure, and function of the intestinal microbial communities of P. monodon from two different regions in Guangdong. Our results identified 176 phyla across both populations, with Proteobacteria and Firmicutes being predominant. Furthermore, we identified 3095 genera, with Photobacterium, Vibrio, and Aliiroseovarius being the most dominant. Functional gene analysis based on KEGG data indicated that the carbohydrate metabolism and amino acid metabolism were significant at the secondary metabolic pathway level. The eggNOG functional annotation revealed that the genes involved in replication, recombination, and repair are of paramount importance. The CAZy annotation results indicated that Glycoside Hydrolases (GH) have the highest abundance. The Pfam annotation analysis showed that the two most prevalent domains are P-loop NTPase and NADP Rossmann. Our investigation provides a reference for species-level and functional-level analyses of the intestinal microbiota of P. monodon, contributing valuable insights into its microbial ecology.

Reproductive Tract Microbial Transitions from Late Gestation to Early Postpartum Using 16S rRNA Metagenetic Profiling in First-Pregnancy Heifers.

Studies in recent years indicate that reproductive tract microbial communities are crucial for shaping mammals' health and reproductive outcomes. Following parturition, uterine bacterial contamination often occurs due to the open cervix, which may lead to postpartum uterine inflammatory diseases, especially in primiparous individuals. However, investigations into spatio-temporal microbial transitions in the reproductive tract of primigravid females remain limited. Our objective was to describe and compare the microbial community compositions in the vagina at late gestation and in the vagina and uterus at early postpartum in first-pregnancy heifers. Three swab samples were collected from 33 first-pregnancy Holstein Friesian heifers: one vaginal sample at gestation day 258 ± 4, and vaginal and uterine samples at postpartum day 7 ± 2. Each sample underwent 16S rRNA V4 region metagenetic analysis via Illumina MiSeq, with bioinformatics following Mothur MiSeq SOP. The reproductive tract bacterial communities were assigned to 1255 genus-level OTUs across 30 phyla. Dominant phyla, accounting for approximately 90% of the communities, included Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes, and Fusobacteria. However, the results revealed distinct shifts in microbial composition between the prepartum vagina (Vag-pre), postpartum vagina (Vag-post), and postpartum uterus (Utr-post). The Vag-pre and Utr-post microbial profiles were the most distinct. The Utr-post group had lower relative abundances of Proteobacteria but higher abundances of Bacteroidetes, Fusobacteria, and Tenericutes compared to Vag-pre, while Vag-post displayed intermediate values for these phyla, suggesting a transitional profile. Additionally, the Utr-post group exhibited lower bacterial richness and diversity compared to both Vag-pre and Vag-post. The unsupervised probabilistic Dirichlet Multinomial Mixtures model identified two distinct community types: most Vag-pre samples clustered into one type and Utr-post samples into another, while Vag-post samples were distributed evenly between the two. LEfSe analysis revealed distinct microbial profiles at the genus level. Overall, specific microbial markers were associated with anatomical and temporal transitions, revealing a dynamic microbial landscape during the first pregnancy and parturition. These differences highlight the complexity of these ecosystems and open new avenues for research in reproductive biology and microbial ecology.

Variation in Bacterial and Fungal Communities in Soils from Three Major Apple Pear (Pyrus bretschneideri Rehd.) Orchards.

Microbial communities are closely related to the overall health and quality of soil, but studies on microbial ecology in apple pear orchard soils are limited. In the current study, 28 soil samples were collected from three apple pear orchards, and the composition and structure of fungal and bacterial communities were investigated by high-throughput sequencing. The molecular ecological network showed that the keystone taxa of bacterial communities were Actinobacteria, Proteobacteria, Gemmatimonadetes, Acidobacteria, Nitrospirae, and Chloroflexi, and the keystone taxon of fungal communities was Ascomycota. Mantel tests showed that soil texture and pH were important factors shaping soil bacterial and fungal communities, and soil water soluble organic carbon (WSOC) and nitrate nitrogen (NO3--N) were also closely related to soil bacterial communities. Canonical correspondence analysis (CCA) and variation partition analysis (VPA) revealed that geographic distance, soil texture, pH, and other soil properties could explain 10.55%, 13.5%, and 19.03% of the overall variation in bacterial communities, and 11.61%, 13.03%, and 20.26% of the overall variation in fungal communities, respectively. The keystone taxa of bacterial and fungal communities in apple pear orchard soils and their strong correlation with soil properties could provide useful clues toward sustainable management of orchards.

Viral Diversity and Ecological Impact of DNA Viruses in Dominant Tick Species in China.

Ticks are blood-feeding ectoparasites that also transmit various pathogens, posing severe risks to human and animal health. DNA viruses play a crucial role in the microbial ecology of ticks, but their distribution and ecological significance remain largely undetermined. Here, we assembled an extensive catalog encompassing 4320 viral operational taxonomic units (vOTUs) from six main dominant tick species in China, of which 94.8% have not been found in any other environment. To bridge the knowledge gap in tick DNA virus research and provide a crucial resource platform, we developed the Tick DNA Virus Database. This database includes the vOTUs that are known to cause diseases. Most of the predicted vOTUs are associated with dominant bacterial and archaeal phyla. We identified 105 virus-encoded putative auxiliary metabolic genes (AMGs) that are involved in host metabolism and environmental adaptation, potentially influencing ticks through both top-down and bottom-up mechanisms. The identification of microbial communities and antibiotic resistance in wild tick species suggests that wild ticks are reservoirs of antibiotic resistance and potential spreaders of antibiotic resistance. These findings reveal the potential role of tick viruses in ecosystems, highlighting the importance of monitoring tick microbiomes to address global public health challenges.

Microbial diversity, aflatoxin M1 contamination and potential lactic acid starters for commercially produced traditional fermented dairy beverages, a case of Bongo from Uganda

This was a cross sectional study that examined the microbial ecology and aflatoxin M1 contamination in commercially produced Bongo, a traditionally fermented dairy beverage from Uganda. It also involved isolation and preliminary evaluation of defined starter cultures for Bongo. Microbial analyses were done by agar plating. Microbial identification was initially done by biochemical testing and microscopy. Followed by 16S rRNA and Internal Transcribed Spacer (ITS) region sequencing for bacteria and yeasts, respectively. Community DNA was studied using Next Generation Sequencing (NGS). Aflatoxin M1 was determined using Ultra High Pressure Liquid Chromatography - Triple Quadrupole Mass Spectrometry. (GTG)5 Rep PCR fingerprinting was used to cluster candidates for starter culture evaluation. The rates of acidification of cow milk by candidate LAB coupled with sensory acceptability of the milk they fermented were the criteria for the selection of the starter cultures. Results indicated that lactic acid bacteria (LAB) and yeasts were the dominant groups. Molds, coliforms, enterococci and staphylococci were also detected. Aflatoxin M1 were insignificant. Based on agar plating technique, lactococci and lactobacilli were dominant, however, NGS showed Streptococci. Agar plating also showed Saccharomyces as dominant, but NGS showed Dipodascus. Cluster analysis identified 18 LAB, which were evaluated for starter culture properties. Three showed promise suggesting that they could be used for commercial production of cultured milks like Bongo.

Activated sludge microbial community assembly: the role of influent microbial community immigration.

Wastewater treatment plants (WWTPs) are host to diverse microbial communities and receive a constant influx of microbes from influent wastewater. However, the impact of immigrants on the structure and activities of the activated sludge (AS) microbial community remains unclear. To gain insight on this phenomenon known as perpetual community coalescence, the current study utilized controlled manipulative experiments that decoupled the influent wastewater composition from the microbial populations to reveal the fundamental mechanisms involved in immigration between sewers and AS-WWTP. The immigration dynamics of heterotrophs were analyzed by harvesting wastewater biomass solids from three different sewer systems and adding to synthetic wastewater. Immigrating influent populations were observed to contribute up to 14% of the sequencing reads in the AS. By modeling the net growth rate of taxa, it was revealed that immigrants primarily exhibited low or negative net growth rates. By developing a protocol to reproducibly grow AS-WWTP communities in the lab, we have laid down the foundational principles for the testing of operational factors creating community variations with low noise and appropriate replication. Understanding the processes that drive microbial community diversity and assembly is a key question in microbial ecology. In the future, this knowledge can be used to manipulate the structure of microbial communities and improve system performance in WWTPs.IMPORTANCEIn biological wastewater treatment processes, the microbial community composition is essential in the performance and stability of the system. This study developed a reproducible protocol to investigate the impact of influent immigration (or perpetual coalescence of the sewer and activated sludge communities) with appropriate reproducibility and controls, allowing intrinsic definitions of core and immigrant populations to be established. The method developed herein will allow sequential manipulative experiments to be performed to test specific hypothesis and optimize wastewater treatment processes to meet new treatment goals.

Microbial assemblages and associated biogeochemical processes in Lake Bonney, a permanently ice-covered lake in the McMurdo Dry Valleys, Antarctica

BackgroundLake Bonney, which is divided into a west lobe (WLB) and an east lobe (ELB), is a perennially ice-covered lake located in the McMurdo Dry Valleys of Antarctica. Despite previous reports on the microbial community dynamics of ice-covered lakes in this region, there is a paucity of information on the relationship between microbial genomic diversity and associated nutrient cycling. Here, we applied gene- and genome-centric approaches to investigate the microbial ecology and reconstruct microbial metabolic potential along the depth gradient in Lake Bonney.ResultsLake Bonney is strongly chemically stratified with three distinct redox zones, yielding different microbial niches. Our genome enabled approach revealed that in the sunlit and relatively freshwater epilimnion, oxygenic photosynthetic production by the cyanobacterium Pseudanabaena and a diversity of protists and microalgae may provide new organic carbon to the environment. CO-oxidizing bacteria, such as Acidimicrobiales, Nanopelagicales, and Burkholderiaceae were also prominent in the epilimnion and their ability to oxidize carbon monoxide to carbon dioxide may serve as a supplementary energy conservation strategy. In the more saline metalimnion of ELB, an accumulation of inorganic nitrogen and phosphorus supports photosynthesis despite relatively low light levels. Conversely, in WLB the release of organic rich subglacial discharge from Taylor Glacier into WLB would be implicated in the possible high abundance of heterotrophs supported by increased potential for glycolysis, beta-oxidation, and glycoside hydrolase and may contribute to the growth of iron reducers in the dark and extremely saline hypolimnion of WLB. The suboxic and subzero temperature zones beneath the metalimnia in both lobes supported microorganisms capable of utilizing reduced nitrogens and sulfurs as electron donors. Heterotrophs, including nitrate reducing sulfur oxidizing bacteria, such as Acidimicrobiales (MAG72) and Salinisphaeraceae (MAG109), and denitrifying bacteria, such as Gracilimonas (MAG7), Acidimicrobiales (MAG72) and Salinisphaeraceae (MAG109), dominated the hypolimnion of WLB, whereas the environmental harshness of the hypolimnion of ELB was supported by the relatively low in metabolic potential, as well as the abundance of halophile Halomonas and endospore-forming Virgibacillus.ConclusionsThe vertical distribution of microbially driven C, N and S cycling genes/pathways in Lake Bonney reveals the importance of geochemical gradients to microbial diversity and biogeochemical cycles with the vertical water column.

Long-term legacy of phytoremediation on plant succession and soil microbial communities in petroleum-contaminated sub-Arctic soils

Abstract. Phytoremediation can be a cost-effective method of restoring contaminated soils using plants and associated microorganisms. Most studies follow the impacts of phytoremediation solely across the treatment period and have not explored long-term ecological effects. In 1995, a phytoremediation study was initiated near Fairbanks, Alaska, to determine how the introduction of annual grasses and/or fertilizer would influence degradation of petroleum hydrocarbons (PHCs). After 1 year, grass and/or fertilizer-treated soils showed greater decreases in PHC concentrations compared to untreated plots. The site was then left for 15 years with no active site management. In 2011, we re-examined the site to explore the legacy of phytoremediation on contaminant disappearance, as well as on plant and soil microbial ecology. We found that the recruited vegetation and the current bulk soil microbial community structure and functioning were all heavily influenced by initial phytoremediation treatment. The number of diesel-degrading microorganisms (DDMs) was positively correlated with the percentage cover of vegetation at the site, which was influenced by initial treatment. Even 15 years later, the initial use of fertilizer had significant effects on microbial biomass, community structure, and activity. We conclude that phytoremediation treatment has long-term, legacy effects on the plant community, which, in turn, impact microbial community structure and functioning. It is therefore important to consider phytoremediation strategies that not only influence site remediation rates in the short-term but also prime the site for the restoration of vegetation over the long-term.