Construction and characterization of a genome-scale ordered mutant collection of Bacteroides thetaiotaomicron

Genomic analyses have revealed how the gut microbiota impacts human health. However, knowledge about the physiology of most gut commensals is largely lacking. Here, we sorted cells from a pooled library to construct an ordered collection of transposon-insertion mutants in the model commensal Bacteroides thetaiotaomicron. We applied a pooling strategy with barcode sequencing to locate mutants and created a condensed collection with single insertions in 2,565 genes. This effort enabled the development of an accurate model for progenitor-collection assembly, which identified strain-abundance biases and multi-insertion strains as important factors that limit coverage. To demonstrate the potential for phenotypic screening, we analyzed growth dynamics and morphology of the condensed collection and identified growth defects and altered cell shape in the sphingolipid-synthesis gene BT0870 and the thiamine-scavenging gene BT2397. Analyses of this collection and utilization of the platform described herein to construct future ordered libraries will increase understanding of gut commensal physiology and colonization strategies.

Unlocking the hydrolytic mechanism of GH92 α-1,2-mannosidases: computation inspires using C-glycosides as Michaelis complex mimics

The conformational changes in a sugar moiety along the hydrolytic pathway are key to understand the mechanism of glycoside hydrolases (GHs) and to design new inhibitors. The two predominant itineraries for mannosidases go via OS2  B2,5  1S5 and 3S  3H4  1C4. For the CAZy family 92, the conformational itinerary was unknown. Published complexes of Bacteroides thetaiotaomicron GH92 catalyst with a S-glycoside and mannoimidazole indicate a 4C1  4H5/1S5  1S5 mechanism. However, as observed with the GH125 family, S-glycosides may not act always as good mimics of GH’s natural substrate. Here we present a cooperative study between computations and experiments where our results predict the E5  B2,5/1S5  1S5 pathway for GH92 enzymes. Furthermore, we demonstrate the Michaelis complex mimicry of a new kind of C-disaccharides, whose biochemical applicability was still a chimera.

Oregano Essential Oils Promote Rumen Digestive Ability by Modulating Epithelial Development and Microbiota Composition in Beef Cattle

This study aimed to explore the effects of oregano essential oils (OEO) on the rumen digestive ability using multi-omics sequencing techniques. Twenty-seven castrated Pingliang red cattle were randomly separated into three groups (3 cattle/pen; n = 9) and fed on a daily basal diet supplemented with 0 (Con group), 130 mg (L group), and 260 mg (H group) OEO. The finishing trial lasted for 390 days, and all cattle were slaughtered to collect rumen tissue and content samples. We found that the rumen papillae length in the H group was higher than in the Con group. Amylase concentrations were decreased in the H group than the Con group, whereas the β-glucosidase and cellulase concentrations increased. Compared to the Con group, the relative abundance of propionate and butyrate in the H group was significantly higher. Higher relative abundance of Parabacteroides distasonis and Bacteroides thetaiotaomicron were observed with increasing OEO concentration. The function of rumen microbiota was enriched in the GH43_17 family, mainly encoding xylanase. Besides, metabolites, including heparin, pantetheine, sorbic acid, aspirin, and farnesene concentrations increased with increasing OEO dose. A positive correlation was observed between Parabacteroides distasonis, Bacteroides thetaiotaomicron, and β-glucosidase, cellulase and propionate. The abundance of Parabacteroides distasonis and Parabacteroides_sp._CAG:409 were positively correlated with sorbic acid and farnesene. In summary, OEO supplementation increased the rumen digestive ability by modulating epithelial development and microbiota composition in beef cattle. This study provides a comprehensive insight into the OEO application as an alternative strategy to improve ruminant health production.

The First Report of Bacteroides fragilis and Bacteroides thetaiotaomicron Relative Abundance in Obese Iranian Population

Background and Objectives: Obesity is a multifactorial disorder, and gut microbiota has a fundamental role in its pathophysiology. Bacteroides spp. has significant roles in gut microbiota- host interactions that determine health and disease development. Since the gut microbiota pattern changes based on different criteria in each population, we studied the abundance of two important Bacteroides strains, Bacteroides fragilis, and Bacteroides thetaiotaomicron, in Iranian obese and normal-weight subjects for the first time. Methods: In this study, 100 participants were recruited and classified based on their body mass index (BMI). The subjects were divided into normal (average BMI, 22.37 kg/m2) and obese (average BMI, 29.10 kg/m2) groups. Bacterial DNA was extracted from the samples, and quantitative polymerase chain reaction (qPCR) was conducted based on 16s rDNA universal primers. Finally, the correlation between bacterial abundance and obesity was investigated. Results: The results of qPCR showed that the relative abundance means of B. fragilis in normal weight and obese subjects was 8.68 × 1012 and 9.27 × 1012 cfu/mL, respectively. Also, the relative abundance mean of B. thetaiotaomicron in normal weight and obese subjects was 2.32 × 1012 and 5.39 × 1012 cfu/mL, respectively. Although obese subjects had more B. fragilis and B. thetaiotaomicron abundance compared to subjects with normal weight, no significant difference was identified between relative abundance of B. fragilis (P = 0.79) and B. thetaiotaomicron (P = 0.18) in the two groups. Conclusions: Although obese subjects had more B. fragilis and B. thetaiotaomicron abundance compared to normal-weight subjects, no significant difference was identified between the two groups. Since Bacteroides spp. have significant role in gut microbiota-host interaction, determination of their abundance in obesity development and targeting restoration of gut microbiota pattern could be valuable in controlling obesity. In this regard, dietary intervention could be based on determination of gut microbiota pattern in certain populations.