Abundant Operational Taxonomic Units Research Articles

Strength Lies in Diversity: How Community Diversity Limits Salmonella Abundance in the Chicken Intestine.

The transfer of the intestinal microbiota from adult to juvenile animals reduces Salmonella prevalence and abundance. The mechanism behind this exclusion is unknown, however, certain member species may exclude or promote pathogen colonization and Salmonella abundance in chickens correlates with intestinal community composition. In this study, newly hatched chicks were colonized with Salmonella Typhimurium and 16S rRNA libraries were generated from the cecal bacterial community at 21, 28, 35, and 42 days of age. Salmonella was quantified by real-time PCR. Operational taxonomic units (OTUs) were assigned, and taxonomic assignments were made, using the Ribosomal Database Project. Bacterial diversity was inversely proportional to the Salmonella abundance in the chicken cecum (p < 0.01). In addition, cecal communities with no detectable Salmonella (exclusive community) displayed an increase in the abundance of OTUs related to specific clostridial families (Ruminococcaceae, Eubacteriaceae, and Oscillospiraceae), genera (Faecalibacterium and Turicibacter) and member species (Ethanoligenens harbinense, Oscillibacter ruminantium, and Faecalibacterium prausnitzii). For cecal communities with high Salmonella abundance (permissive community), there was a positive correlation with the presence of unclassified Lachnospiraceae, clostridial genera Blautia and clostridial species Roseburia hominis, Eubacterium biforme, and Robinsoniella peoriensis. These findings strongly support the link between the intestinal bacterial species diversity and the presence of specific member species with Salmonella abundance in the chicken ceca. Exclusive bacterial species could prove effective as direct-fed microbials for reducing Salmonella in poultry while permissive species could be used to predict which birds will be super-shedders.

High-throughput sequencing-based analysis of bacterial communities associated with Barbour's seahorses (Hippocampus barbouri) from Surigao del Norte, Philippines.

This study identified the bacterial community composition associated with Barbour's seahorses (Hippocampus barbouri). Seahorses and samples from the surrounding environment (sediment and water) were collected from Cantiasay Island in Surigao del Norte, Philippines. Genomic DNA was isolated from these samples, and the V1-V3 region of the 16S rRNA gene was amplified and sequenced on the Illumina MiSeq platform. There were 929803 sequence reads corresponding to 859 operational taxonomic units (OTUs), which were obtained from the six genomic libraries. The results showed that the most abundant OTUs were affiliated to the phylum Proteobacteria, particularly those belonging to the genera Shewanella, Pseudomonas, Acinetobacter and Aeromonas, which were found to be dominant in seahorse gut samples. The only exception was for the skin of male seahorses, which was dominated by members belonging to the genus Bacillus (phylum Firmicutes). These findings on bacterial community structure and composition may shed light on therapeutic claims of Barbour's seahorses and help in the conservation of this species.

Microbial colonization and community stability in a marine post-smolt RAS inoculated with a commercial starter culture

The performance of a commercial starter culture was investigated in a new marine post-smolt RAS, analyzing the microbial communities of 100 samples collected monthly over a year from biofilter biofilm carriers, tank wall biofilm, production water and fish skin. Totally 139 operational taxonomic units (OTUs) were defined in the starter culture, of which the classified members of Rhodobacterales, Bacteroidetes, Alteromonadales and Planctomycetes were largely the first colonizers of the biofilter carriers. Early colonizing OTUs that dominated biofilter biofilm carriers (> 5% relative abundance) were stably present over time, but the development went slowly from a few OTUs with very high relative abundance to several dominant ones with lower relative abundance. Operating taxonomic units not associated with the starting culture became prominent on the biofilter biofilm carriers only towards the end of the trial period. These were termed environmental OTUs. Comparing the two OTU quantitives in a ratio, where counts were based on all OTUs in the sample, the starter culture OTUs:environmental OTUs were 1.2 and 0.9 at the first and last sampling time for the biofilter biofilm carriers. Correspondingly, for all defined OTUs in the RAS sampling sites together, the ratio changed from 0.8 to 0.6 during experiment. Independent of origin, omniscient OTUs at a sampling site, did also have the highest relative abundances and were normally shared between biofilter biofilm carriers and the production water. New and lost OTUs between sampling times were on average 44 % of the OTUs defined, and this OTUflow was strongest for low abundant environmental OTUs. The maturation of the biofilter with respect to nitrification took long time, and the Nitrospira strain in the starter culture was not adapted to marine salinities. Still, we report a controlled colonization of the marine RAS by the starter culture.

Microbiome Signature and Diversity Profiling of Normal Skin of Human in Saudi Arabia

Background: Studying human skin-associated bacterial communities is crucial to understanding human diseases, disease progression, and their role in maintaining human health. Objectives: This study aimed to identify normal (healthy) skin microbiome signatures of eight individuals living in Jeddah, Makkah Al-Mukarramah region, Saudi Arabia. Methods: The study involved the analysis of resident skin microbiome in inner elbow of the right arm after ethical approval is issued and an informed consent form is signed by participant individuals. Results: Phylogenetic tree indicated the existence of four phyla, e.g., Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria. Firmicutes were shown to be the highest in abundance, while Bacteroidetes were the lowest. At the genus level, Staphylococcus was the highest in abundance, while Enterococcus was the lowest. At the species level, Bacillus cereus was the highest in abundance, while Roseomonas mucosa was the lowest. The analysis for the highly abundant operational taxonomic units (OTUs) indicated a dramatic difference between sexes referring to either genera or species of which Staphylococcus sp., Erwinia sp., Pseudomonas sp., Sphingomonas sp., Corynebacterium sp., Propionibacterium acnes, Kocuria palustris are higher in males, while Bacillus cereus, Bacillus sp., Erwinia sp., Corynebacterium sp., Micrococcus sp., Pseudomonas sp. are lower in males. Conclusions: The study succeeded in detecting the skin microbiome of individuals in Saudi Arabia.

Comparison of gut microbiota in autism spectrum disorders and neurotypical boys in China: A case-control study

BackgroundAutism spectrum disorders (ASDs) are a set of complex neurobiological disorders. Growing evidence has shown that the microbiota that resides in the gut can modulate brain development via the gut–brain axis. However, direct clinical evidence of the role of the microbiota–gut–brain axis in ASD is relatively limited. MethodsA case-control study of 71 boys with ASD and 18 neurotypical controls was conducted at China-Japan Friendship Hospital. Demographic information and fecal samples were collected, and the gut microbiome was evaluated and compared by 16S ribosomal RNA gene sequencing and metagenomic sequencing. ResultsA higher abundance of operational taxonomic units (OTUs) based on fecal bacterial profiling was observed in the ASD group. Significantly different microbiome profiles were observed between the two groups. At the genus level, we observed a decrease in the relative abundance of Escherichia, Shigella, Veillonella, Akkermansia, Provindencia, Dialister, Bifidobacterium, Streptococcus, Ruminococcaceae UCG_002, Megasphaera, Eubacterium_coprostanol, Citrobacter, Ruminiclostridium_5, and Ruminiclostridium_6 in the ASD cohort, while Eisenbergiella, Klebsiella, Faecalibacterium, and Blautia were significantly increased. Ten bacterial strains were selected for clinical discrimination between those with ASD and the neurotypical controls. The highest AUC value of the model was 0.947. ConclusionSignificant differences were observed in the composition of the gut microbiome between boys with ASD and neurotypical controls. These findings contribute to the knowledge of the alteration of the gut microbiome in ASD patients, which opens the possibility for early identification of this disease.

109 Is the equine gut colonised in utero by amniotic bacteria?

Until very recently it was believed that the uterus was sterile, with microbial colonisation of the foal's gastrointestinal tract (GIT) commencing at birth. GIT colonisation is an integral developmental process, critical for an optimal functioning immune system and ongoing health of the foal. The in utero colonisation paradigm has now challenged this concept, indicating that the amniotic fluid (AF) composition contains non-pathogenic microbiota that potentially commence colonisation of the GIT during the process of fetal swallowing. In turn, preparing the foal for life outside the womb. This study examined the presence of microbiota in the AF of healthy pregnant mares, identifying microorganisms that have the potential to colonise the fetal GIT in utero. AF samples were obtained from 14 healthy mares in the second stage of parturition, before the rupture of the amnion. When the amnion was clearly visible, the surface was seared with a sterile spatula and AF extracted with a sterile needle and syringe. The sample had no contact with any other surface of the mare and was immediately snap-frozen and stored at −80°C for microbiota analysis. Genomic DNA was extracted and processed for 16S rRNA gene amplicon sequencing, followed by bioinformatic analysis of the results using QIIME2 and CalypsoV8.84 software packages. Microbiota was detected in all AF samples. Communities were of low diversity, as indicated by the α diversity parameters Chao1, Evenness, Richness and Simpson's Index. Operational Taxonomic Units (OTUs) of the top 20 taxa showed some host variation. The most abundant OTUs across the majority of samples were classified to the phyla Opisthokonta (100% of samples), Bacteroidetes (79%), Actinobacteria (57%) and Firmicutes (57%). At the family level, Staphylococcacea (79%) and Microbacteriaceae (71%) were the most abundant. These microbes have been previously detected across other locations of the equine microbiome, including the oral, esophageal, distal GIT and vaginal ecosystems. Our results have demonstrated the presence of a complex microbial ecosystem of low diversity in the AF of healthy equine pregnancies, likely of maternal origin. The environmental and gastrointestinal microbes detected in the AF displayed similarities to other equine ecosystems, suggesting microbial translocation from the mare to the AF. Further research would be required to determine if these AF microbes are being ingested by the fetus, thus commencing GIT colonisation in utero . Deciphering microbial colonisation of the foal's GIT has important implications for maternal management before parturition and could lead to the development of strategies to enhance foal health outcomes.

Assessing soil microbial biodiversity as affected by grazing and woody vegetation cover in a temperate savannah

AbstractAlthough it is well established that land use and management affects soil microbial biodiversity, with consequences for soil‐mediated ecosystem services, knowledge on human‐induced variability in soil bacterial and fungal communities is more abundant for cropping than for livestock systems. In particular, grazing management affects vegetation cover and light availability to lower vegetation layers, particularly in wooded grazing ecosystems. Our objective was to evaluate the use of T‐RFLP analysis to characterise soil microbial community patterns. We worked in a grazed temperate savannah in Entre Ríos (Argentina) and focused on two farms with different grazing management, resulting in different vegetation cover. At each farm, we sampled soils from four conditions forming a two shrub cover conditions (open and close) by two light availability conditions (high and low) factorial set with three replicates. T‐RFLP data for bacteria and fungi were used to characterise biodiversity patterns (diversity indexes, relative abundance of operational taxonomic units and community structure). Analyses were performed, between and within farms, using nonmetric multidimensional scaling analysis, non‐parametric multivariate analysis of variance and network analysis. Variation patterns of soil microbial communities were detected using T‐RFLP analysis in savannah soils from these two livestock farms. Farm, light and shrub conditions were statistically significant factors shaping variation patterns. For bacterial communities, light availability and shrub cover affected biodiversity and community structure respectively, but this impact was detected only for the farm with lower grazing intensity. For fungal communities, light availability was the factor shaping community structure, also with slightly higher impact on the farm with lower grazing intensity. These results suggest that in temperate woodlands lower grazing intensity, associated with higher vegetation cover, increases bacterial biodiversity while it decreases fungal community structure. Soil bacterial and fungal communities, showed contrasting patterns, and appear as sensitive indicators to monitor grazing management effects. We showed that T‐RFLP analysis is a cost‐effective methodology (compared to Illumina sequencing) that would be useful to disentangle impact of grazing intensity and vegetation cover in grazed temperate savannah.

Ectomycorrhizal fungal communities associated with Larix gemelinii Rupr. in the Great Khingan Mountains, China.

Larix gemelinii is an important tree species in the Great Khingan Mountains in Northeast China with a high economic and ecological value for its role in carbon sequestration and as a source of lumber and nuts. However, the ectomycorrhizal (EM) fungal diversity and community composition of this tree remain largely undefined. We examined EM fungal communities associated with L. gemelinii from three sites in the Great Khingan Mountains using Illumina Miseq to sequence the rDNA ITS2 region and evaluated the impact of spatial, soil, and climatic variables on the EM fungal community. A total of 122 EM fungal operational taxonomic units (OTUs) were identified from 21 pooled-root samples, and the dominant EM fungal lineages were /tricholoma, /tomentella-thelephora, /suillus-rhizopogon, and /piloderma. A high proportion of unique EM fungal OTUs were present; some abundant OTUs largely restricted to specific sites. EM fungal richness and community assembly were significantly correlated with spatial distance and climatic and soil variables, with mean annual temperature being the most important predictor for fungal richness and geographic distance as the largest determinant for community turnover. Our findings indicate that L. gemelinii has a rich and distinctive EM fungal community contributing to our understanding of the montane EM fungal community structure from the perspective of a single host plant that has not been previously reported.

Genetic Diversity in Marine Planktonic Ciliates (Alveolata, Ciliophora) Suggests Distinct Geographical Patterns – Data From Chinese and European Coastal Waters

Unraveling geographic distribution patterns of planktonic protists is a central goal in marine microbial ecology. Using a novel combination of recently developed phylogenetic and network analyses on a V4 18S rDNA metabarcoding dataset, we here analyzed the genetic diversity of marine planktonic ciliate communities in Chinese and European coastal waters. Thereby, our approach provided an unprecedented perspective on geographic patterns inferred from ciliate genetic diversity and accomplished a very fine resolution down to single nucleotides within operational taxonomic units (OTUs). While most OTUs (87%) exclusively contained sequences of either Chinese or European origin, those OTUs detected in both regions comprised the vast majority of reads (84%). Phylogenetic analyses of OTUs belonging to the same taxon revealed genetically distinct clades that were geographically restricted to either Chinese or European coastal waters. The detection of signature nucleotides emphasized this genetic distinction of Chinese and European clades. Second-level clustering of OTUs and reference sequences in two selected taxa (the oligotrichid Spirotontonia and the tintinnid Tintinnidium) revealed the presence of several potentially new species or ones lacking genetic reference data. Geographic patterns were also discovered by network analyses within 700 widespread and abundant OTUs; in 77 of these OTUs, European and Chinese sequences formed significantly assortative groups. These assortative groupings indicated a higher genetic similarity among sequences from the same region than between sequences from different regions. Our results demonstrate that detailed analyses of metabarcoding data down to single nucleotide differences expand our perception of geographical distribution patterns and provide insights into historic and ongoing effective dispersal in protists. The congruent discovery of geographic patterns at different levels of resolution (between and within OTUs) suggests that cosmopolitan distribution in marine planktonic ciliates is less common than previously postulated.

Dynamics of microbial diversity during the composting of agricultural straw

The dynamic changes in microbial diversity during the aerobic composting of agricultural crop straw with additives were evaluated using high-throughput sequencing at four phases of composting (mesophilic, thermophilic, cooling and maturation phases). In addition, the physicochemical parameters of the composting system were determined in this study. The fermentation time of the thermophilic period was prolonged with the addition of urea or urea combined with a microbial agent. The ratio of C/N and germination index variation indicated that the additives were favorable for composting, because the additives directly changed the physicochemical properties of the compost and had effects on the diversity and abundance of bacteria and fungi. The abundance of operational taxonomic units (OTUs), diversity index (Shannon) and richness index (Chao1) of fungi and bacteria were found to significantly increase when urea+microbial agents were added to straw in the thermophilic phase. The relative abundance of the predominant bacteria and fungi at the phylum and genus levels differed during different composting phases. The abundance of the phyla Firmicutes and Proteobacteria declined in the order of treatments SNW>SN>S (S is straw only compost; SN is straw+5 kg t−1 urea compost; and SNW is straw+5 kg t−1 urea+ 1 kg t−1 microbial agent compost) in the thermophilic phase. The abundance of the genera Staphylococcus, Bacillus and Thermobifida followed the same order in the mesophilic phase. Ascomycota accounted for more than 92% of the total fungal sequences. With the progression of the composting process, the abundance of Ascomycota decreased gradually. The abundance of Ascomycota followed the order of S>SN>SNW during the thermophilic phase. The abundance of Aspergillus accounted for 4–59% of the total abundance of fungi and increased during the first two sampling periods. Aspergillus abundance followed the order of SNW>SN>S. Additionally, principal component analysis (PCA) revealed that the community compositions in the straw and straw+urea treatments were similar, and that the bacterial communities in treatments S, SN and SNW in the mesophilic phase (at day 1) were different from those observed in three other phases (at days 5, 11, and 19, respectively), while the fungal communities showed only slight variations in their structure in response to changes in the composting process. Canonical correlation analysis (CCA) and redundancy analysis (RDA) showed that total carbon (TC), NO3−-N (NN), electrical conductivity (EC) and pH were highly correlated with community composition. Therefore, this study highlights that the additives are beneficial to straw composting and result in good quality compost.

Gut microbial differences in breast and prostate cancer cases from two randomised controlled trials compared to matched cancer-free controls.

Implicated in several chronic diseases, the gastrointestinal microbiome is hypothesised to influence carcinogenesis. We compared faecal microbiota of newly diagnosed treatment-naïve overweight and obese cancer patients and matched controls. Cases were enrolled in presurgical weight-loss trials for breast (NCT02224807) and prostate (NCT01886677) cancers and had a body mass index (BMI) ≥25 kg/m2. Cancer-free controls were matched 1:1 by age (±5 years), race, gender, and BMI (±5 kg/m2). All participants provided faecal samples; isolated bacterial DNA were PCR amplified at the V4 region of the 16S rRNA gene and analysed using the QIIME pipeline. Tests compared cases versus controls, then separately by gender. Microbial alpha-diversity and beta-diversity were assessed, and relative abundance of Operational Taxonomic Units (OTU's) were compared at the genus level, with false discovery rate (FDR) correction. 22 overweight and obese cancer patients were matched with 22 cancer-free controls, with an average BMI of 30.5±4.3 kg/m2, age 54.4±5.3 years, and 54.5% were black. Fourteen matches were made between breast cancer cases and healthy female controls, and 8 matches were made with prostate cancer cases and healthy male controls. Comparison of all cases and controls revealed no differences in alpha diversity, though prostate cancer patients had higher Chao1 (P=0.006) and Observed Species (P=0.036) than cancer-free males. Beta-diversity metrics were significantly different between cases and controls (P<0.03 for all tests in whole sample and in men), though only unweighted Unifrac was different in women (P=0.005). Kruskal Wallis tests indicated significant differences among 16 genera in all matches, 9 in female, and 51 in male. This study suggests the faecal microbiota of treatment-naive breast and prostate cancer patients differs from controls, though larger samples are needed to substantiate these findings. Trial registration: NIH Clinical Trials, NCT01886677, NCT02224807, registered 26 June 2013, 25 Aug 2014 (respectively) - retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT01886677; https://clinicaltrials.gov/ct2/show/NCT02224807.

Xylanase Supplementation Modulates the Microbiota of the Large Intestine of Pigs Fed Corn-Based Fiber by Means of a Stimbiotic Mechanism of Action.

This research tested the hypothesis that xylanase modulates microbial communities within the large intestine of growing pigs fed corn-based fiber through a stimbiotic mechanism(s) of action (MOA). Sixty gilts were blocked by initial body weight, individually housed, and randomly assigned to one of four dietary treatments (n = 15): a low-fiber (LF) control, a high-fiber (HF) control containing 30% corn bran, HF+100 mg/kg xylanase (HF+XY), and HF+50 mg/kg arabinoxylan-oligosaccharide (HF+AX). Pigs were fed dietary treatments for 46 days. On day 46, pigs were euthanized, and mucosa and lumen contents were collected from the cecum and the colon. The V4 region of 16S rRNA genes was sequenced and clustered into 5,889, 4,657, 2,822, and 4,516 operational taxonomic units (OTUs), in the cecal contents and mucosa and colonic contents and mucosa, respectively. In cecal contents, HF+XY increased measures of α-diversity compared to LF (p < 0.001). Relative to LF, HF increased the prevalence of 44, 36, 26, and 8, and decreased 19, 9, 21, and 10, of the 200 most abundant OTUs from the cecal contents and mucosa and colonic contents and mucosa, respectively (Q < 0.05). Compared to LF, HF increased the abundance of OTUs from the Treponema_2, Ruminococcus_1 genera, from the Lachnospiraceae, Ruminococcaceae, and Prevotellaceae families. In contrast, relative to LF, HF decreased Turicibacter and Lactobacillus in the cecal contents, and Megasphaera and Streptococcus in the mucosa. Relative to HF, HF+XY increased 32, 16, 29, and 19 and decreased 27, 11, 15, and 10 of the 200 most abundant OTUs from the cecal contents and mucosa and colonic contents and mucosa, respectively (Q < 0.05). The addition of xylanase to HF further increased the abundance of OTUs from the Lachnospiraceae and Ruminococcaceae families across the large intestine. Compared to HF, HF+XY increased the abundance of Lactobacillus, Bifidobacterium, and Faecalibacterium among all locations (Q < 0.05). However, HF+AX did not increase the prevalence of these genera in the large intestine. Supplementing xylanase to HF increased hidden-state predictions of microbial enzymes associated with arabinoxylan degradation, xylose metabolism, and short-chain fatty acid production. These data suggest xylanase elicits a stimbiotic MOA in the large intestine of pigs fed corn-based fiber.

Fungal Diversity and Community Assembly of Ectomycorrhizal Fungi Associated With Five Pine Species in Inner Mongolia, China.

Ectomycorrhizal (EM) fungi play vital roles in ensuring host plants’ health, plant diversity, and the functionality of the ecosystem. However, EM fungal diversity, community composition, and underlying assembly processes in Inner Mongolia, China, where forests are typically semiarid and cold-temperate zones, attract less attention. In this study, we investigated EM fungal communities from 63 root samples of five common pine plants in Inner Mongolia across 1,900 km using Illumina Miseq sequencing of the fungal internal transcribed spacer 2 region. We evaluated the impact of host plant phylogeny, soil, climatic, and spatial variables on EM fungal diversity and community turnover. Deterministic vs. stochastic processes for EM fungal community assembly were quantified using β-nearest taxon index scores. In total, we identified 288 EM fungal operational taxonomic units (OTUs) belonging to 31 lineages, of which the most abundant lineages were Tomentella–Thelephora, Wilcoxina, Tricholoma, and Suillus–Rhizopogon. Variations in EM fungal OTU richness and community composition were significantly predicted by host phylogeny, soil (total nitrogen, phosphorus, nitrogen–phosphorus ratio, and magnesium), climate, and spatial distance, with the host plant being the most important factor. β-nearest taxon index demonstrated that both deterministic and stochastic processes jointly determined the community assembly of EM fungi, with the predominance of stochastic processes. At the Saihanwula site selected for preference analysis, all plant species (100%) presented significant preferences for EM fungi, 54% of abundant EM fungal OTUs showed significant preferences for host plants, and 26% of pairs of plant species and abundant fungal OTUs exhibited remarkably strong preferences. Overall, we inferred that the high diversity and distinctive community composition of EM fungi associated with natural pine species in Inner Mongolia and the stochastic processes prevailed in determining the community assembly of EM fungi. Our study shed light on the diversity and community assembly of EM fungi associated with common pine species in semiarid and cold temperate forests in Inner Mongolia, China, for the first time and provided a better understanding of the ecological processes underlying the community assembly of mutualistic fungi.

Microbial Interactions as Drivers of a Nitrification Process in a Chemostat.

This article deals with the inclusion of microbial ecology measurements such as abundances of operational taxonomic units in bioprocess modelling. The first part presents the mathematical analysis of a model that may be framed within the class of Lotka–Volterra models fitted to experimental data in a chemostat setting where a nitrification process was operated for over 500 days. The limitations and the insights of such an approach are discussed. In the second part, the use of an optimal tracking technique (developed within the framework of control theory) for the integration of data from genetic sequencing in chemostat models is presented. The optimal tracking revisits the data used in the aforementioned chemostat setting. The resulting model is an explanatory model, not a predictive one, it is able to reconstruct the different forms of nitrogen in the reactor by using the abundances of the operational taxonomic units, providing some insights into the growth rate of microbes in a complex community.

Ammonium Removal in Aquaponics Indicates Participation of Comammox Nitrospira

Aquaponic systems are sustainable solutions for food production combining fish growth (aquaculture) and production of vegetables (hydroponic) in one recirculating system. In aquaponics, nitrogen-enriched wastewater from fish in the aquaculture serves as fertilizer for the plants in the hydroponics, while the nitrogen-depleted and detoxified water flows back to the aquaculture. To investigate bacterial nitrogen-cycling in such an aquaponic system, measurements of nitrogen species were coupled with time-resolved 16S rRNA gene profiling and the functional capacity of organisms was studied using metagenomics. The aquaponic system was consistently removing ammonia and nitrite below 23 µM and 19 µM, and nitrate to steady-state concentrations of about 0.5 mM. 16S rRNA gene amplicon sequencing of sediments exposed in the pump sump revealed that typical signatures of canonical ammonia-oxidising microorganisms were below detection limit. However, one of the most abundant operational taxonomic units (OTU) was classified as a member of the genus Nitrospira with a relative abundance of 3.8%. For this genus, also genome scaffolds were recovered encoding the only ammonia monooxygenase genes identified in the metagenome. This study indicates that even in highly efficient aquaponic systems, comammox Nitrospira were found to participate in ammonium removal at low steady-state ammonia concentrations.

Effect of organic and conventional farming on soil bacterial diversity of pecan tree (Carya illinoensis K. Kosh) orchard across two phenological stages.

We described the bacterial diversity of walnut grove soils under organic and conventional farming. The bacterial communities of rhizospheric and nonrhizospheric soils of pecan tree (Carya illinoensis K. Koch) were compared considering two phenological stages (sprouting and ripening). Sixteen operational taxonomic units (OTUs) were identified significantly more abundant according to the plant development, only one according to the farming condition, and none according to the soil origin. The OTUs specificaly abundant according to plant development included Actinobateria (2) and Betaproteobacteria (1) related OTUs more abundant at the sprouting stage, while at the fruit ripening (FR) stage the more abundant OTUs were related to Actinobacteria (6), Alphaproteobacteria (6), and unclassified Bacteria (1). The Gaiellaceae OTU18 (Actinobacteria) was more abundant under conventional farming. Thus, our study revealed that the plant development stage was the main factor shaping the bacterial community structure, while less influence was noticed for the farming condition. The bacterial communities exhibited specific metabolic capacities, a large range of carbon sources being used at the FR stage. The identified OTUs specifically more abundant represent indicators providing useful information on soil condition, potential tools for the management of soil bacterial communities.

A vertically and temporally diverse bacterial community in a shallow lake-water sediment site of a eutrophic lake

ABSTRACT Aquatic sediment accumulates organic matter and provides microenvironments for the growth of a diverse microbial community with key roles in the biogeochemical nutrient cycling of the lacustrine ecosystem. We investigated vertical and temporal variations of the sedimentary bacterial community composition in a shallow coastal site of eutrophic Lake Pamvotis (northwestern Greece) across 3 horizons between the surface and 5 cm below lake floor (cmblf) during transition to the cold period of the year (Oct–Dec). For this analysis we used Illumina sequencing of the V3–V4 hypervariable regions of the 16S rRNA gene. Bacterial community composition was dominated by Proteobacteria, Bacteroidetes, and Chloroflexi. Rare operational taxonomic units (OTUs) represented 96.09% (standard deviation 0.61%) of richness per sample, contributing significantly to diversity, and only a few became abundant during the study. Abundant OTUs represented <0.005% of richness but accounted for 20–33% of reads in different samples, mainly associated with anaerobic/microaerobic taxa common in polluted aquatic systems, sludge digesters, or wastewater (e.g., Thiolamprovum pedioforme, Candidatus Competibacter, Petrimonas). About 50% of OTUs found at the surface persisted with depth while shared OTUs between any 2 months were always >70% of richness. Overall, sediment bacterial community composition was diverse but remained largely homogeneous in the sediment of this human-impacted ecosystem at the temporal and vertical scale studied here. The study revealed the potential of specific bacterial taxa (e.g., Anaerolineae, Bellilinea) to serve as monitors of specific pollutants (e.g., polycyclic aromatic hydrocarbons) as well as the need to investigate the role of the rare biosphere in ecosystem functioning.

Xylanase modulates the microbiota of ileal mucosa and digesta of pigs fed corn-based arabinoxylans likely through both a stimbiotic and prebiotic mechanism.

The experimental objective was to characterize the impact of insoluble corn-based fiber, xylanase, and an arabinoxylan-oligosaccharide on ileal digesta and mucosa microbiome of pigs. Three replicates of 20 gilts were blocked by initial body weight, individually-housed, and assigned to 1 of 4 dietary treatments: a low-fiber control (LF), a 30% corn bran high-fiber control (HF), HF+100 mg/kg xylanase (HF+XY), and HF+50 mg/kg arabinoxylan oligosaccharide (HF+AX). Gilts were fed their respective treatments for 46 days. On day 46, pigs were euthanized and ileal digesta and mucosa were collected. The V4 region of the 16S rRNA was amplified and sequenced, generating a total of 2,413,572 and 1,739,013 high-quality sequences from the digesta and mucosa, respectively. Sequences were classified into 1,538 mucosa and 2,495 digesta operational taxonomic units (OTU). Hidden-state predictions of 25 enzymes were made using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States 2 (PICRUST2). Compared to LF, HF increased Erysipelotrichaceae_UCG-002, and Turicibacter in the digesta, Lachnospiraceae_unclassified in the mucosa, and decreased Actinobacillus in both (Q<0.05). Relative to HF, HF+XY increased 19 and 14 of the 100 most abundant OTUs characterized from digesta and mucosa, respectively (Q<0.05). Notably, HF+XY increased the OTU_23_Faecalibacterium by nearly 6 log2-fold change, compared to HF. Relative to HF, HF+XY increased genera Bifidobacterium, and Lactobacillus, and decreased Streptococcus and Turicibacter in digesta (Q<0.05), and increased Bifidobacterium and decreased Escherichia-Shigella in the mucosa (Q<0.05). Compared to HF, HF+AX increased 5 and 6 of the 100 most abundant OTUs characterized from digesta and mucosa, respectively, (Q<0.05), but HF+AX did not modulate similar taxa as HF+XY. The PICRUST2 predictions revealed HF+XY increased gene-predictions for enzymes associated with arabinoxylan degradation and xylose metabolism in the digesta, and increased enzymes related to short-chain fatty acid production in the mucosa. Collectively, these data suggest xylanase elicits a stimbiotic and prebiotic mechanism.

Effect of xylanase and xylo-oligosaccharide supplementation on growth performance and faecal bacterial community composition in growing pigs

This study was conducted to investigate the effects of xylanase (XYL) and xylo-oligosaccharide (XOS) supplementation on the growth performance and faecal bacterial community composition in growing pigs over time. In this 35-day trial, a total of 464 grower pigs with an average initial body weight (BW) of 14.5 kg (SD ±1.56 kg) were blocked into mixed sexed pens of 4–5 pigs balanced for BW, sex and litter origin. Pens were randomly allocated to 1 of 4 dietary treatments in a 2 × 2 factorial treatment arrangement with 2 concentrations of supplementary XYL (0 and 0.15 g/kg) and XOS (0 and 0.20 g/kg). On a weekly basis, pen feed intake and pigs were weighed to calculate pig performance. Faecal samples from 32 male pigs were collected on days 1, 14 and 35 of the trial and analysed to profile the bacterial communities through 16S ribosomal ribonucleic acid (rRNA) sequencing of the V4 region on the MiSeq platform (Illumina). There was no effect of XYL on pig performance, nor was there an interaction between XYL and XOS. Pigs supplemented with XOS had a poorer gain to feed ratio during the first week (Day 1–7) of the trial (P < 0.01). During the second week of the trial (Day 8–14), pigs fed XOS showed an improved average daily gain compared to those without XOS (P < 0.01), but there were no performance effects in the overall trial period (Day 1–35). Alpha diversity increased over time (P < 0.05), and as an index of bacterial community compositions, beta diversity also changed over time (P < 0.05), but there was no overall effect of treatment on alpha or beta diversity. Despite no overall treatment effect, certain operational taxonomic units (OTUs) associated with Muribaculaceae_ge and Prevotellaceae NK3B31 group were higher in all 3 dietary treatments compared to the unsupplemented control diet (P < 0.05). In summary, XYL and XOS had limited effect on pig performance in this trial. Faecal bacterial communities significantly changed over time but despite influencing certain OTUs, treatment had no overall effect on faecal bacterial community composition. Supplementation of XYL or XOS, individually or simultaneously, increased the abundance of OTUs belonging to the Muribaculaceae and Prevotellaceae families which are associated with carbohydrate metabolism, indicating that these bacteria are likely involved in the mechanistic pathways of XYL and XOS.

Microbiomic profiles of bile in patients with benign and malignant pancreaticobiliary disease.

417 Background: In pancreaticobiliary (PB) cancers, there is a paucity of data on predictive and pathophysiologic role of the biliary microbiome. We analyzed bile collected from patients with benign and malignant PB diseases to identify microbiomic signatures associated with malignancy. Methods: We collected bile samples from consenting patients during routine endoscopic retrograde cholangiopancreatography at the Cleveland Clinic, approved by the Institutional Review Board. DNA was extracted from bile specimens using PowerViral RNA/DNA Isolation kit. Bacterial 16S rRNA gene amplification and library construction were performed according to the 16S Metagenomic Sequencing Library Preparation guide from Illumina. Post-sequencing analysis was done using QIIME (Quantitative Insights Into Microbial Ecology), Bioconductor phyloseq, microbiomeSeq and mixMC packages. Results: Of 46 enrolled patients, 32 had PB cancers including pancreatic (N = 25), cholangiocarcinoma (N = 6), and gallbladder (N = 1). The rest (N = 14) had benign PB diseases including acute and chronic pancreatitis, and gallstones. Using multivariate approach in mixMC to classify Operational Taxonomic Units (OTUs), we found a predominance of genera Dicekeya (p = 0.0002), [ Eubacterium] hallii group (p = 0.0007) , Bacteroides (p = 0.00099) , Faecalibacterium (p = 0.007) , Facklamia (p = 0.013) , Peptococcus (p = 0.013) , Bergeyella (p = 0.0024) , Lachnospira (p = 0.026) , and Lactobacillus (p = 0.025) in bile samples from PB cancers as compared to benign PB diseases. Furthermore, bile samples from patients with pancreatic cancer showed an increased abundance of genera Enterobacter, Parabacteroides, Atopobium, Alloprevotella, Prevotella 7, Acinetobacter, Bergeyella, Clostridium sensu stricto, Lactobacillus, and Rothia; and a decreased abundance of genera Tannerella, Peptococcus, Colinsella, Capnocytophaga, Achromobacter, Ruminococcus 2, Bacteroides, Alistipes, Barnesiella,, Lachnoclostridium, Lautropia, Akkermansia, and Christensenellaceae R-7 group as compared to bile samples from patients with cholangiocarcinoma. Conclusions: Distinct microbiome signatures are associated with benign and malignant PB diseases. There is a difference in the relative abundance of OTUs in bile samples between patients with benign PB diseases vs PB cancers, and between pancreatic cancer vs cholangiocarcinoma. Our findings raise the possibility that either these OTUs have a role in carcinogenesis, or that changes in the microenvironment of benign PB diseases differ from PB cancers leading to distinct separation of the OTU clusters. Further studies to explore and validate our findings are needed.