Observed Operational Taxonomic Units Research Articles

Associations of Sedentary Behavior and Screen Time with Human Gut Microbiome Composition and Diversity.

Human gut microbiome richness, diversity, and composition are associated with physical activity and impaired glycemic control; however, the associations with sedentary behavior and screen time are not as well-established. This study evaluated associations of sedentary behavior and screen time with the alpha diversity and composition of the human gut microbiome in adults with and without impaired glycemic control. Sedentary behavior and screen time data were collected via survey from 47 adults (38% with impaired glycemic control). Microbiome composition and alpha diversity were determined in fecal microbial DNA. Sedentary behavior was negatively associated with the number of observed operational taxonomic units (OTUs), Chao 1 Index, and Fisher's Alpha Index. These associations were slightly attenuated but remained significant when controlling for covariates. Screen time was negatively associated with the number of observed OTUs, Shannon Index, and Fisher's Alpha Index; however, only the association with observed OTUs was independent of all covariates. Our findings suggest sedentary behavior and screen time may be significant influencers of compositional changes in human gut microbiota. This may be a potential mechanism linking sedentary behavior and screen time to an increased risk of type 2 diabetes.

Biochar Mitigates the Negative Effects of Microplastics on Sugarcane Growth by Altering Soil Nutrients and Microbial Community Structure and Function.

Microplastic pollution in sugarcane areas of China is severe, and reducing the ecological risks is critical. Biochar has been widely used in soil remediation. This study aims to explore the effects and mechanisms of microplastics combined with or without biochar on sugarcane biomass, soil biochemical properties in red soil through a potted experiment. The results show that, compared with control (CK), treatments with microplastics alone reduced the dry biomass of sugarcane, soil pH, and nitrogen (N) and phosphorus (P) contents by an average of 8.8%, 2.1%, 1.1%, and 2.0%, respectively. Interestingly, microplastics combined with biochar could alleviate the negative effects of microplastic accumulation on sugarcane growth and soil quality. There were significant differences in the bacterial community alpha diversity indices and compositions among different treatments. Compared with CK, treatments with microplastics alone obviously decreased the observed operational taxonomic units (OTUs) and the Chao1 and Shannon indices of soil total bacteria (16S rRNA gene-based bacteria) while increasing them in phoD-harboring bacteria. Microplastics combined with biochar treatments significantly increased the abundance of Subgroup_10 for the 16S rRNA gene and treatments with microplastics alone significantly increased the relative abundance of Streptomyces for the phoD gene compared to CK. Moreover, compared with microplastics alone, the treatments with microplastics combined with biochar increased the relative abundance of Subgroup_10, Bacillus, Pseudomonas in soil total bacteria, and Amycolatopsis and Bradyrhizobium in phoD-harboring bacteria, most of which can inhibit harmful bacteria and promote plant growth. Additionally, different treatments also changed the abundance of potential microbial functional genes. Compared to CK, other treatments increased the abundance of aerobic ammonia oxidation and denitrification but decreased the abundance of nitrate respiration and nitrogen respiration; meanwhile, these four functional genes involved in N cycling processes were obviously higher in treatments with microplastics combined with biochar than in treatments with microplastics alone. In conclusion, microplastics combined with biochar could alleviate the negative effects of microplastic accumulation on sugarcane biomass by altering soil nutrients and microbial community structure and function.

Rhodotorula benthica culture as an alternative to antibiotics improves growth performance by improving nutrients digestibility and intestinal morphology, and modulating gut microbiota of weaned piglets.

The effects of Rhodotorula benthica culture (RBC) and antibiotics (AB) on the growth performance, nutrients digestibility, morphological indicators, and colonic microbiota of weaning piglets were explored. Ninety-six (Duroc × Landrace × Large) weaned piglets (21-day-old) weighing 7.7 ± 0.83 kg, were randomly allocated to 4 dietary treatments. They were fed with basal diet (CON), basal diet + 25 mg/kg bacitracin zinc + 5 mg/kg colistin sulfate (AB), 5 g/kg reduction in soybean meal of basal diet + 5 g/kg RBC (RBC1), or 10 g/kg reduction in soybean meal of basal diet + 10 g/kg RBC (RBC2). The results showed that dietary RBC1 improved the body gain/feed intake (G/F) of weaned piglets than the CON diet, and the RBC2 diet improved the average daily gain and G/F than CON and AB diets from days 15 to 28 (P < 0.05). Supplementation of RBC2 improved the apparent total tract digestibility of dry matter, nitrogen, and gross energy in weaned piglets compared to controls from days 15 to 28 (P < 0.05). Dietary AB, RBC1, and RBC2 enhanced the ileal villus height (VH) and VH/crypt depth (CD), and these two indicators were greater in the RBC2-treated piglets than in the AB- and RBC1-treated piglets (P < 0.05). The activity of serum superoxide dismutase (SOD) was enhanced by dietary AB, RBC1, and RBC2 (P < 0.05). Serum glutathione (GSH) concentration was elevated by dietary RBC1 and RBC2 (P < 0.05). According to 16S rRNA sequence analysis, AB- and RBC2-treated piglets had a higher relative abundance of Firmicutes and Lachnospiraceae in the colon digesta, and more abundant Lactobacillus was found in RBC1-treated piglets, as compared to the CON group. Additionally, RBC2 supplementation increased the α diversity [Chao1, PD-whole-tree, and observed operational taxonomic units (OTUs)] compared to the CON group. Taken together, the dietary RBC improved the growth performance of weaned piglets. In addition, 10 g/kg of RBC2 in the diet achieved better effects on higher ADG, ileal villi morphology, and stronger antioxidant capacity than dietary AB and RBC1 in weaning piglets.

Host species and habitats shape the bacterial community of gut microbiota of three non-human primates: Siamangs, white-handed gibbons, and Bornean orangutans.

The gut microbiome is essential for a host to digest food, maintain health, and adapt to environments. Bacterial communities of gut microbiota are influenced by diverse factors including host physiology and the environment. Many non-human primates (NHPs), which are physiologically close to humans, are in danger of extinction. In this study, the community structure of the gut microbiota in three NHPs: siamangs (Symphalangus syndactylus, Ss), Bornean orangutans (Pongo pygmaeus, Pp), and white-handed gibbons (Hylobates lar, Hl)—housed at the largest Zoo in Taiwan were analyzed. Pp and Ss were housed in the Asian tropical rainforest area, while Hl was housed in two separate areas, the Asian tropical rainforest area and the conservation area. Bacterial community diversity of Ss, indicated by the Shannon index, was significantly higher compared with that of Hl and Pp, while the richness (Chao 1) and observed operational taxonomic units (OTUs) were similar across the three species of NHPs. Host species was the dominant factor shaping the gut microbial community structure. Beta-diversity analysis including non-metric multidimensional scaling (NMDS) and unweighted pair group method with arithmetic mean (UPGMA) suggested gut bacterial communities of Hl housed in the conservation area were closely related to each other, while the bacterial communities of Hl in the rainforest area were dispersedly positioned. Further analysis revealed significantly higher abundances of Lactobacillus fermentum, L. murinus, and an unclassified species of Lactobacillus, and a lower abundance of Escherichia-Shigella in Hl from the conservation area relative to the rainforest area. The ratio of Lactobacillus to Escherichia-Shigella was 489.35 and 0.013 in Hl inhabiting the conservation and rainforest areas, respectively. High abundances of Lactobacillus and Bifidobacterium and a high ratio of Lactobacillus to Escherichia-Shigella were also observed in one siamang with notable longevity of 53 years. Data from the study reveal that host species acted as the fundamental driving factor in modulating the community structure of gut microbiota, but that habitats also acted as key determinants within species. The presence and high abundance of probiotics, such as Bifidobacterium and Lactobacillus, provide potential indicators for future diet and habitat optimization for NHPs, especially in zoological settings.

Effect of commercial slow-release urea product on in vitro rumen fermentation and ruminal microbial community using RUSITEC technique

BackgroundThe objectives of this study were to determine the effect of commercial slow-release urea (SRU) on in vitro fermentation characteristics, nutrient digestibility, gas production, microbial protein synthesis and bacterial community using a rumen simulation technique (RUSITEC). The experiment was a completely randomized design with four treatments and four replications of each treatment. Treatments were: control diet (no SRU addition), control diet plus 0.28% SRU (U28), or plus 0.56% SRU (U56), and control diet that was modified substituting a part of soybean meal equivalent to 0.35% SRU (MU35; dry matter [DM] basis). The experiment consisted of 8 d of adaptation and 7 d of data and sample collection. Rumen inoculum was obtained from three ruminally fistulated Angus cows fed the same diet to the substrate incubated.ResultsDigestibility of DM, organic matter (OM), crude protein (CP), fibre and starch was not affected, but daily production of gas (P < 0.07) and methane (P < 0.05) was quadratically increased with increasing SRU supplementation. The increase of SRU addition did not affect fermentation pH and total volatile fatty acid (VFA) production, whereas linearly (P < 0.01) decreased proportion of propionate, and linearly (P < 0.01) increased acetate to propionate ratio and ammonia nitrogen (N) concentration. The microbial N efficiency was also linearly (P < 0.03) improved with increasing supplementation of SRU. In comparison with control diet, the dietary substitution of SRU for part of soybean meal increased (P < 0.05) the digestibility of DM, OM and CP and decreased (P < 0.02) the total gas production. The total VFA production and acetate to propionate ratio did not differ between control and MU35, whereas the proportion of butyrate was lower (P < 0.05) and that of branched-chain VFA was greater (P < 0.05) with MU35 than control diet. Total and liquid-associated microbial N production as well as ammonia N concentration were greater (P < 0.03) with MU35 than control diet. Observed operational taxonomic units (OTUs), Shannon diversity index, and beta diversity of the microbial community did not differ among treatments. Taxonomic analysis revealed no effect of adding SRU on the relative abundance of bacteria at the phylum level, while at the genus level, the beneficial impact of SRU on relative abundance of Rikenellaceae and Prevotellaceae in feed particle-associated bacteria, and the abundance of Roseburia in liquid associate bacteria was greater (P < 0.05) with MU35.ConclusionsSupplementation of a dairy cow diet with SRU showed potential of increase in ammonia N concentration and microbial protein production, and change fermentation pattern to more acetate production. Adding SRU in dairy cow diet also showed beneficial effect on improving digestibility of OM and fibre. The results suggest that SRU can partially substitute soybean meal in dairy cow diet to increase microbial protein production without impairing rumen fermentation.

Effects of Growth Stage and Rearing Pattern on Pig Gut Microbiota

Understanding the developmental period or the patterns of gut microbiota is important for nutritionists when designing a feed formula or adjusting a feeding strategy. The effects of growth stage or rearing pattern on pig gut microbiota have not been fully investigated. Herein, 39 fecal samples from pigs aged 3-9months under two rearing patterns were collected to analyze the gut microbiome. Samples were clustered into three distinct groups, namely, early (3months), middle (5months), and late (7 and 9months) stages, using principal coordinate analysis and analysis of similarities test. The rearing-pattern effects were very minimal, and no differences were observed in the alpha diversity [observed operational taxonomic units (OTUs) and Shannon index] of gut microbiota. From early and middle to late stage, Shannon index gradually decreased and OTUs gradually increased. Pigs at early stage were enriched with bacteria from family Prevotellaceae, including the genera Prevotella_9 and Prevotellaceae_NK3B31, whereas pigs at late stage were enriched with family Ruminococcaceae, including genera Ruminococcaceae_UCG-005 and Oscillospira. Pigs in the semi-free-grazing farm group were significantly enriched with bacteria from order Clostridiales. Growth stage better explained the changes in porcine gut microbiota than rearing patterns.

Gut Microbiota From Sjögren syndrome Patients Causes Decreased T Regulatory Cells in the Lymphoid Organs and Desiccation-Induced Corneal Barrier Disruption in Mice.

Sjögren syndrome (SS) is an autoimmune inflammatory disorder characterized by secretory dysfunction in the eye and mouth; in the eye, this results in tear film instability, reduced tear production, and corneal barrier disruption. A growing number of studies show that homeostasis of the ocular surface is impacted by the intestinal microbiome, and several 16S sequencing studies have demonstrated dysbiosis of the intestinal microbiota in SS patients. In this study, we utilized metagenomic sequencing to perform a deeper analysis of the intestinal microbiome using stools collected from sex- and age-matched healthy (n = 20), dry eye (n = 4) and SS (n = 7) subjects. The observed Operational Taxonomic Units (OTUs) and Shannon alpha diversity were significantly decreased in SS compared to healthy controls, and there was a significant inverse correlation between observed OTUs and ocular severity score. We also identified specific bacterial strains that are differentially modulated in SS vs. healthy subjects. To investigate if the differential composition of intestinal microbiome would have an impact on the immune and eye phenotype, we performed functional studies using germ-free mice colonized with human intestinal microbiota from SS patients and healthy controls. Flow cytometry analysis demonstrated reduced frequency of CD4+ FOXP3+ cells in ocular draining cervical lymph nodes (CLN) in mice colonized with SS patient intestinal microbiota 4 weeks post-colonization. We also found that offspring of SS-humanized mice also have fewer CD4+FOXP3+ cells in the CLN as well as spleen, demonstrating vertical transmission. SS-humanized mice subjected to desiccating stress exhibited greater corneal barrier disruption as compared to healthy control humanized mice under the same conditions. Taken together, these data support the hypothesis that the intestinal microbiota can modulate ocular surface health, possibly by influencing development of CD4+ FOXP3+ regulatory T cells (Tregs) in the ocular draining lymph nodes.

The Gut Microbiome, Seleno-Compounds, and Acute Myocardial Infarction.

Background: Gut microbiome alterations might be considered a metabolic disorder. However, the relationship between the microbiome and acute myocardial infarction (AMI) has not been properly validated. Methods: The feces of 44 subjects (AMI: 19; control: 25) were collected for fecal genomic DNA extraction. The variable region V3–V4 of the 16S rRNA gene was sequenced using the Illumina MiSeq platform. The metabolite amounts were analyzed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathways. Results: The bacteria were more enriched in the AMI group both in the observed operational taxonomic units (OTUs) and faith phylogenetic diversity (PD) (p-value = 0.01 and <0.001 with 95% CI, individually). The Selenomonadales were less enriched in the AMI group at the family, genus, and species levels (all linear discriminant analysis (LDA) scores > 2). Seleno-compounds were more abundant in the AMI group at the family, genus, and species levels (all LDA scores > 2). Conclusions: This is the first study to demonstrate the association of Selenomonadales and seleno-compounds with the occurrence of AMI. Our findings provide an opportunity to identify a novel approach to prevent and treat AMI.

Responses of bacterial taxonomic attributes to mercury species in rhizosphere paddy soil under natural sulphur-rich biochar amendment

Biochar and sulphur (S) are important factors regulating the level, speciation and transformation of mercury (Hg), leading to alterations in the assemblage of the soil microbial community. However, variations in the taxonomic attributes of the rhizosphere soil bacterial community arising from the Hg speciation in paddy soil, amended with natural S-rich biochar (NSBC) derived from the pyrolysis of S-rich oilseed rape straw, remain unclear. Herein, a rice pot experiment was conducted. Hg-polluted paddy soils were amended with NSBC and low-S biochar (LSBC) to evaluate the role of Hg chemical form affected by NSBC in regulating the taxonomic attributes of rhizosphere soil, including microbial abundance, composition, and ecological clusters within the co-occurrence network of microbial communities. Results showed that microbial abundance was higher in soils with lower Hg levels, and mean increases of 149 observed operational taxonomic units (OTUs) and 238 predicted OTUs (Chao 1) were observed, with a 1 mg kg−1 decrease in the total Hg (T-Hg) content. Among the 13 predictor variables, the T-Hg content was the strongest and most consistent predictor of the bacterial taxonomic attributes. This finding may be attributed to the fact that the drastic reduction in T-Hg and Hg bioavailability induced by NSBC results in the decrease of Hg stress on the soil microbiome. Moreover, NSBC amendment shifted the ecological clusters toward the amelioration of Hg pollution.

Microbial community alteration in tongue squamous cell carcinoma.

Tongue squamous cell carcinoma (TSCC) is the most common oral cavity malignancy. The role of the microbial community in TSCC development and progression is unclear. In the present study, 23 patients with TSCC were recruited. Tissue DNA was extracted from cancer and paracancerous normal tissues from all participants. Next-generation 16S rDNA amplicon sequencing and functional prediction were applied for taxonomic analysis. Alpha diversity measurements using the Shannon and Simpson diversity indexes indicated a significant increase in the microbiotic diversity of cancer samples (Shannon index: P = 0.001, Simpson index: P = 0.015); otherwise, no differences were found when using observed operational taxonomic units (OTUs) and Chao1 index (observed OTUs: P = 0.261, Chao1 index: P = 0.054). The dominant phyla of the microbiota included Bacteroidetes, Proteobacteria, Firmicutes, Actinobacteria, and Fusobacteria. Multivariate analysis of variance (Adonis) and nonparametric analysis of similarities (ANOSIM) based on unweighted unifrac distances demonstrated differences in the bacterial community structure between the two groups (P = 0.001 for Adonis, P = 0.001 for ANOSIM). Compared with the normal samples, Neisseria, Streptococcus, and Actinomyces levels decreased significantly in cancer samples. Co-occurrence network analysis implied that the bacterial community in cancer was more conserved than that in normal tissue. Matched-pair analysis of cancer and control samples revealed a significant alteration in the relative abundance of specific taxa. These findings will enrich our knowledge of the association between the oral microbial community and TSCC. Further experiments should investigate the potential carcinogenic mechanism of microbial community alterations in TSCC. KEY POINTS: • Microbial community role in tongue squamous cell carcinoma. • Significant alteration of microbiome found between cancer and normal tissues. • Microbial community alteration and potential carcinogenic mechanism.

Oral microbiome in Proliferative Verrucous Leukoplakia exhibits loss of diversity and enrichment of pathogens

ObjectivesOral microbiome plays an important role in oral diseases. Among them, proliferative verrucous leucoplakia (PVL) is an uncommon form of progressive multifocal leukoplakia with a worryingly rate of malignant transformation. Here, we aimed to characterize the oral microbiome of PVL patients and compare it with those of healthy controls. Material and methodsOral biopsies from ten PVL patients and five healthy individuals were obtained and used to compare their microbial communities. The sequence of the V3-V4 region of 16S rRNA gene was used as the taxonomic basis to estimate and analyze the composition and diversity of bacterial populations present in the samples. ResultsOur results show that the oral microbial composition and diversity are significantly different among PVL patients and healthy donors. The average number of observed operational taxonomic units (OTUs) was higher for healthy donors than for PVL, proving a loss of diversity in PVL. Several OTUs were found to be more abundant in either group. Among those that were significantly enriched in PVL patients, potential protumorigenic pathogens like Oribacterium sp. oral taxon 108, Campylobacter jejuni, uncultured Eubacterium sp., Tannerella, and Porphyromonas were identified. ConclusionOral microbiome dysbiosis was found in patients suffering from PVL. To the best of our knowledge, this is the first study investigating the oral microbiome alterations in PVL and, due to the limited number of participants, additional studies are needed. Oral microbiota-based biomarkers may be helpful in predicting the risks for the development of PVL.

Herbs and Spices Modulate Gut Microbiota Composition: A Randomized, Crossover, Controlled-Feeding Study

Abstract Objectives The effect of culinary doses of herbs and spices, consumed as part of a well-defined dietary pattern, on gut microbiota composition has not been previously studied. The aim of this study was to examine gut microbial composition following an average American diet (carbohydrate: 50% kcal; protein: 17%; total fat: 33%; saturated fat: 11%; sodium: 3000 mg/d) containing herbs and spices at 0.5 (Low Spice Diet; LSD), 3.3 (Moderate Spice Diet; MSD) and 6.6 (High Spice Diet; HSD) g/day/2100 kcal in adults at-risk for cardiovascular disease (CVD). Methods Fifty-four adults (57% female; 45 ± 11 years, BMI 29.8 ± 2.9 kg/m2; waist circumference 102.8 ± 7.1 cm) were included in this three-period, randomized, crossover, controlled-feeding study. Each diet was provided for 4-weeks with a minimum 2-week wash-out period. At baseline and the end of each diet period subjects provided a fecal sample for 16S rRNA gene sequencing. QIIME2 was used for data filtration, sequence clustering, taxonomy assignment and statistical analysis. Linear discriminant analysis Effect Size (LEfSe) analyses were used to determine enriched biomarker taxa following each diet. Results No between-diet differences in alpha diversity were observed based on Faith's Phylogenetic Diversity, Observed Operational Taxonomic Units (OTUs), and Pielou's Evenness (Kruskal-Walllis, P &amp;gt; 0.05). Principal coordinate analysis showed beta-diversity following the HSD significantly differed from baseline (PERMANOVA, P = 0.030). LEfSe pairwise analyses identified multiple taxa that were differentially abundant following the spice-containing diets vs. baseline. Compared to baseline, 34 taxa were enriched with the HSD, 22 taxa were enriched with the MSD and 20 taxa were enriched with the LSD. Members of the taxa enriched with the HSD are known short-chain fatty acid producers (Faecalibacterium and Butyricimonas). Conclusions The addition of herbs and spices at 0.5, 3.3, and 6.6 g/day/2100 kcal to an average American diet altered gut microbiota composition, and resulted in enrichment of short-chain fatty acid producers, after 4-weeks in adults at elevated risk for CVD. Funding Sources McCormick Science Institute; NCATS NIH (1UL1TR002014–01); Grant from the Howard Hughes Medical Institute through the Precollege and Undergraduate Science Education Program to Juniata College supported the computational research.

Environmental Influences on the Human Gut Microbiota: A Longitudinal Pilot Study

ObjectivesUrbanization has reduced environmental microorganism exposure, with most Americans spending over 90% of their time indoors. However, gardening remains a viable means of exposure to soil microorganisms and harvesting of edible produce. Accordingly, we aimed to determine relations between gardening, dietary habits, and gut microbiota. MethodsGardening families (N = 10) and non-gardening (control) families (N = 9) were enrolled in a longitudinal pilot trial. Families included two adults and a child (5–18 years) for a total sample size of 54 participants. Fecal samples were collected prior to and at the end of the gardening season. Garden soil samples (n = 9) were collected prior to and at the end of the season. Diet history questionnaires were collected at the beginning and end of the study to measure Healthy Eating Index (HEI) scores. Fecal and soil DNA were extracted, sequenced (V4 region of 16S rDNA gene), and analyzed using DADA2 and QIIME2. Alpha diversity measures were assessed, including Faith’s phylogenetic diversity (PD) and observed operational taxonomic units (OTUs). ResultsGardening families had significantly more fecal OTUs compared to control families (172.3 ± 44.2 vs. 157.0 ± 44.2, respectively; P = 0.03). Gardening families had greater (P = 0.02) Faith’s PD scores and tended (P = 0.08) to have more fecal OTUs than the control group at peak gardening season. In the gardening families, fecal OTUs and Faith’s PD were numerically but not statistically greater at the end of the season compared to baseline (all p’s > 0.05). Prior to the gardening season, gardening adults had greater HEI scores compared to control families (57 ± 9.1 vs. 49 ± 8.8, P = 0.03). HEI scores were not different between groups at the end of the study. ConclusionsThis study revealed that the fecal microbiota of families that garden differs from non-gardening families, and there are detectable changes in the fecal microbial community of gardeners and their family members over the course of the gardening season. Further research is needed to understand the role of diet in these changes and if microbes within the soil move between the soil and gastrointestinal environments. Funding SourcesThis research was funded by the Christopher Family Foundation Food and Family Grant Program.

Compositional Differences in the Oral Microbiome of E-cigarette Users.

Electronic (e)-cigarettes have been advocated as a safer alternative to conventional tobacco cigarettes. However, there is a paucity of data regarding the impact of e-cigarette aerosol deposition on the human oral microbiome, a key component in human health and disease. We aimed to fill this knowledge gap through a comparative analysis of the microbial community profiles from e-cigarette users and healthy controls [non-smokers/non-vapers (NSNV)]. Moreover, we sought to determine whether e-cigarette aerosol exposure from vaping induces persistent changes in the oral microbiome. To accomplish this, salivary and buccal mucosa samples were collected from e-cigarette users and NSNV controls, with additional oral samples collected from e-cigarette users after 2 weeks of decreased use. Total DNA was extracted from all samples and subjected to PCR amplification and sequencing of the V3-V4 hypervariable regions of the 16S rRNA gene. Our analysis revealed several prominent differences associated with vaping, specific to the sample type (i.e., saliva and buccal). In the saliva, e-cigarette users had a significantly higher alpha diversity, observed operational taxonomic units (OTUs) and Faith’s phylogenetic diversity (PD) compared to NSNV controls, which declined with decreased vaping. The buccal mucosa swab samples were marked by a significant shift in beta diversity between e-cigarette users and NSNV controls. There were also significant differences in the relative abundance of several bacterial taxa, with a significant increase in Veillonella and Haemophilus in e-cigarette users. In addition, nasal swabs demonstrated a trend toward higher colonization rates with Staphylococcus aureus in e-cigarette users relative to controls (19 vs. 7.1%; p = n.s.). Overall, these data reveal several notable differences in the oral bacterial community composition and diversity in e-cigarette users as compared to NSNV controls.

Characterization of the core bacteriobiome in the rhizosphere of greenhouse vegetables: taxonomic diversity and putative functions

The aim of the study. The aim was to profile 16S rRNA gene diversity and to assess functional potential of bacterial assemblages in the rhizosphere of some unconventional vegetables grown in protected greenhouse conditions in West Siberia. Location and time of the study. Novosibirsk, Russia, 2016. Methodology. At the end of the growing season in the middle of September the rhizosphere soil was collected from the plants of wax gourd (Benincasa hispida), bitter melon (Momordica charantia), kiwano (Cucumis metuliferus) and cowpea (Vigna unguiculata) grown on peat-based substrate in a polyethylene-protected greenhouse that has been in operation for more than 40 years. The metagenomic DNA was extracted and amplified with V3-V4 primers for 16S rRNA genes, and the amplicons sequenced with Illumina MiSeq. The obtained OTUs tables were used to predict putative functions by running through the FAPROTAX database. Main results. The rhizosphere bacteriobiome was dominated by Proteobacteria (32±11% of the total number of sequence reads), Acidobacteria (23±7%) and Actinobacteria (18±3%) phyla, together accounting for about three quarters of the rhizosphere bacteriobiome. In total 20 bacterial phyla were found. The rhizosphere bacteriobiome was surprisingly diverse with Shannon index ranging 7.0–7.5. The number of the observed operational taxonomic units (OTUs) per sample was very high, ranging 4,500–4,900, and the potential number of OTUs estimated as 5,100–5,700; all those OTUs were evenly and equitably represented in the bacteriobiome, and dominance indices (Simpson dominance and Berger-Parker) were very low. The main dominant OTU represented Bradyrhizobiaceae family and accounted for just 1% on average. Overall the study identified 27 OTUs belonging to the Bradyrhizobiaceae family, but only four of them were ascribed to nitrogen fixation by FAPROTAX. Function prediction by FAPROTAX also suggested that bacteriobiome had a marked potential for the carbon cycle, denitrification, aromatic compound and plant polymer degradation, but no plant pathogens. The biggest difference in rhizosphere bacteriobiome diversity was observed between the bitter melon and the other three vegetable crops: bitter melon had much increased abundance of Arthrobacter and Sphingomonas as compared with wax gourd, kiwano and cowpea, and increased number of bacterial species associated with aromatic compounds degradation. Conclusion. Based on the finding that the studied rhizosphere bacteriobiomes were very diverse, we conclude that the crops were able to recruit diverse microbiota from the peat-based soil substrate, which, in its turn, means that diverse soil substrate microbiota has been sustained over several decades of the greenhouse operation. All crops apparently shaped distinct bacteriobiomes in their rhizosphere, which ideally should be included into studies of plant-associated bacterial diversity profiles for breeding and sustainable production.

Distribution of Bacterial Endophytes in the Non-lesion Tissues of Potato and Their Response to Potato Common Scab

The response of plant endophytes to disease within infected tissues has been well demonstrated, but the corresponding response of endophytes in non-lesion tissues remains unclear. Here, we studied the composition and distribution of bacterial endophytes in potato roots (RE), stems (SE), and tubers (TE), and explored the response of endophytes in non-lesion tissues to potato common scab (PCS), which is a soil-borne disease caused by pathogenic Streptomyces and results in serious losses to the global economy every year. Via high-throughput sequencing, it was seen that the composition of endophytes in roots, stems, and tubers had significant differences (P < 0.05) and the distribution of the bacterial communities illustrated a gradient from soil to root to tuber/stem. PCS significantly reduced bacterial endophytes α-diversity indexes, including ACE and the number of observed operational taxonomic units (OTUs), of RE without significantly reducing the indexes of SE and TE. No significant effect on the composition of endophytes were caused by PCS in roots, tubers, or stems between high PCS severity (H) and low PCS severity (L) infections at the community level, but PCS did have a substantial impact on the relative abundance of several specific endophytes. Rhizobium and Sphingopyxis were significantly enriched in root endophytes with low PCS severity (REL); Delftia and Ochrobactrum were significantly enriched in stem endophytes with low PCS severity (SEL); Pedobacter, Delftia, and Asticcacaulis were significantly enriched in tuber endophytes with high PCS severity (TEH). OTU62, a potential PCS pathogen in this study, was capable of colonizing potato tubers, roots, and stems with few or no symptoms present. Co-occurrence networks showed that the number of correlations to OTU62 was higher than average in these three tissue types, suggesting the importance of OTU62 in endophytic communities. This study clarified the distribution and composition of potato endophytes in tubers, roots, and stems, and demonstrated the response of endophytes in non-lesion tissues to PCS.

The Association of Microbiota and Acute Myocardial Infarction

Background: There is an increasing interest about the mechanisms of acute myocardial infarction (AMI). To investigate the influence of the gut microbiota and the metabolic pathway in associated with AMI, we conduct this case-control study. Methods: We explore 44 subjects with/without AMI (AMI: 19 vs control: 25) and feces are collected from each subject. The fecal microbial genomic DNA is extracted and the variable region, V3–V4, of the 16S rRNA gene is sequencing by the platform of Illumina Miseq. The number of observed operational taxonomic units (OTUs) at 97% similarity level is classified. A histogram of linear discriminant analysis (LDA) distribution is also implemented by the LDA effect size analysis (LEfSe) to find bacteria with significant differences in relative abundance between two groups. The abundances of metabolites are analyzed by the KEGG metabolic pathways. Findings: The abundance of bacteria is more enriched in the AMI group both in the observed OTUs and faith phylogenetic diversity (PD) ( p -value= 0.01 and 2). There is also a difference of the seleno-compound distribution between the groups. The seleno-compound is more abundant in the AMI group at the Family, Genus, and Species levels (all LDA scores >2). Interpretation: This is the first study to demonstrate Selenomonadales and seleno-compound to be associated with the occurrence of AMI. Our findings provided an opportunity for the novel therapeutic strategies for prevention and treatment of AMI. Funding Statement: None. Declaration of Interests: All authors disclose no financial and personal relationships with other people or organisations that could inappropriately influence (bias) their work. Ethics Approval Statement: This study is approved by the Institutional Review Board committee on human research of the medical research department, National Taiwan University Hospital Taiwan.

Dynamics of Fecal Microbiota with and without Invasive Cervical Cancer and Its Application in Early Diagnosis.

Simple SummaryThe fecal microbiome has been suggested to be linked to invasive cervical cancer (ICC). Considering that ICC is common in women, it is important to identify bacterial signatures from fecal microbiota that contribute in classifying cervical cancer. Although previous studies have suggested possible biomarkers based on fecal microbiota, limited information exists in terms of the diagnostic ability using gut microbiota-derived signatures for detecting early ICC. The purpose of this study was to investigate the potential association between early ICC and fecal microbiota and to examine whether fecal microbiota-derived markers can be utilized as a non-invasive tool to diagnose early ICC using machine learning (ML) techniques. Further studies to incorporate quantitative and qualitative characterization of identified individual bacterial genus and validate our model in larger cohorts are imperative in terms of causality for the association between cervical cancer and microbes.The fecal microbiota is being increasingly implicated in the diagnosis of various diseases. However, evidence on changes in the fecal microbiota in invasive cervical cancer (ICC) remains scarce. Here, we aimed to investigate the fecal microbiota of our cohorts, develop a diagnostic model for predicting early ICC, and identify potential fecal microbiota-derived biomarkers using amplicon sequencing data. We obtained fecal samples from 29 healthy women (HC) and 17 women with clinically confirmed early ICC (CAN). Although Shannon’s diversity index was not reached at statistical significance, the Chao1 and Observed operational taxonomic units (OTUs) in fecal microbiota was significantly different between CAN and HC group. Furthermore, there were significant differences in the taxonomic profiles between HC and CAN; Prevotella was significantly more abundant in the CAN group and Clostridium in the HC group. Linear discriminant analysis effect size (LEfSe) analysis was applied to validate the taxonomic differences at the genus level. Furthermore, we identified a set of seven bacterial genera that were used to construct a machine learning (ML)-based classifier model to distinguish CAN from patients with HC. The model had high diagnostic utility (area under the curve [AUC] = 0.913) for predicting early ICC. Our study provides an initial step toward exploring the fecal microbiota and helps clinicians diagnose.

Abstract B04: Gender-based impact of chemo-radiotherapy in the gut microbial diversity and short-chain fatty acid levels of adults with rectal cancers

Abstract Background: The gut microbiome is essential in maintaining the host immune system and may influence the host response to cancer therapeutics. Particularly, by the production of short-chain fatty acids (SCFA) through fermentation of dietary fibers, the gut microbiota helps regulate the immune system. SCFA levels can be affected by changes in gut microbiome composition either by chemo-radiotherapy treatment (CRT) or by gender disparities. We had previously observed that CRT significantly reduced Shannon diversity index and the observed operational taxonomic units (OTUs) during CRT in rectal cancers. Here, we examined the impact of CRT on the levels of SCFAs in adults with RC and investigated gender disparities in the gut microbiome associated with CRT. Methods: We evaluated the gut microbiome of 35 newly diagnosed RC patients scheduled to receive CRT. Demographics, health forms, and stool samples were collected before and after CRT (12-16 treatments). We sequenced the V3-V4 hypervariable region of the 16S rDNA and measured levels of SCFA (from Acetate [C2] to Octonoate [C8]) by LC-MS/MS and results were expressed in log2 (nmol/mg). Alpha diversity (Shannon index [community diversity] and Observed OTUs [community richness]) was calculated using the QIIME2 software and all statistical analyses were performed in R statistical software. Results: Study participants were 35 subjects (20 males vs. 15 females) with an average age of 61.1±9.8 years, even though females tended to be younger than males (57.9±9.8 vs. 63.8±9.3, p=0.08). While females had a trend for a higher Shannon index when compared to males before CRT (6.46±0.31 vs. 6.11±0.43, p=0.09), they had significantly lower Shannon index (5.26±0.31 vs. 5.94±0.60, p=0.02) and observed OTUs (117±7.6 vs. 170±61.0, p=0.03) after CRT, and females had a higher reduction than males (p=0.04). In regard to SCFA, there was an overall increase on the levels of propionate after CRT compared to before CRT (4.10±0.78 vs. 3.64±1.00, p=0.02). However, while there was no difference in the levels of SCFA based on biologic gender after CRT, females had significantly higher levels of propionate (4.32 ±0.68 vs. 3.60±0.90) and butyrate (2.73 ± 0.82 vs. 1.70 ±1.29) at the end of CRT, while men did not show any significant difference in SCFA during CRT. Conclusion: Chemo-radiotherapy resulted in dysbiosis of the gut microbiome and also changed the levels of SCFA. However, these changes in the microbiome associated with CRT were different in females versus males. Our findings suggest that biologic sex may play a key role in the development of personalized cancer treatment strategies. Citation Format: Carlos Sola-Morla, Velda Gonzalez-Mercado, Josue Perez-Santiago. Gender-based impact of chemo-radiotherapy in the gut microbial diversity and short-chain fatty acid levels of adults with rectal cancers [abstract]. In: Proceedings of the AACR Special Conference on the Microbiome, Viruses, and Cancer; 2020 Feb 21-24; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2020;80(8 Suppl):Abstract nr B04.

Microbiomes in the insectivorous bat species Mops condylurus rapidly converge in captivity.

Bats are well known reservoir hosts for RNA and DNA viruses. The use of captive bats in research has intensified over the past decade as researchers aim to examine the virus-reservoir host interface. In this study, we investigated the effects of captivity on the fecal bacterial microbiome of an insectivorous microbat, Mops condylurus, a species that roosts in close proximity to humans and has likely transmitted viral infections to humans. Using amplicon 16S rRNA gene sequencing, we characterized changes in fecal bacterial community composition for individual bats directly at the time of capture and again after six weeks in captivity. We found that microbial community richness by measure of the number of observed operational taxonomic units (OTUs) in bat feces increases in captivity. Importantly, we found the similarity of microbial community structures of fecal microbiomes between different bats to converge during captivity. We propose a six week-acclimatization period prior to carrying out infection studies or other research influenced by the microbiome composition, which may be advantageous to reduce variation in microbiome composition and minimize biological variation inherent to in vivo experimental studies.