Interleukin-22-mediated host glycosylation prevents Clostridioides difficile infection by modulating the metabolic activity of the gut microbiota.

The involvement of host immunity in the gut microbiota-mediated colonization resistance to Clostridioides difficile infection (CDI) is incompletely understood. Here, we show that interleukin (IL)-22, induced by colonization of the gut microbiota, is crucial for the prevention of CDI in human microbiota-associated (HMA) mice. IL-22 signaling in HMA mice regulated host glycosylation, which enabled the growth of succinate-consuming bacteria Phascolarctobacterium spp. within the gut microbiome. Phascolarctobacterium reduced the availability of luminal succinate, a crucial metabolite for the growth of C. difficile, and therefore prevented the growth of C. difficile. IL-22-mediated host N-glycosylation is likely impaired in patients with ulcerative colitis (UC) and renders UC-HMA mice more susceptible to CDI. Transplantation of healthy human-derived microbiota or Phascolarctobacterium reduced luminal succinate levels and restored colonization resistance in UC-HMA mice. IL-22-mediated host glycosylation thus fosters the growth of commensal bacteria that compete with C. difficile for the nutritional niche.

Exposure to Amoxicillin in Early Life Is Associated With Changes in Gut Microbiota and Reduction in Blood Pressure: Findings From a Study on Rat Dams and Offspring.

Background Pediatric hypertension is recognized as an emerging global health concern. Although new guidelines are developed for facilitating clinical management, the reasons for the prevalence of hypertension in children remain unknown. Genetics and environmental factors do not fully account for the growing incidence of pediatric hypertension. Because stable bacterial flora in early life are linked with health outcomes later in life, we hypothesized that reshaping of gut microbiota in early life affects blood pressure (BP) of pediatric subjects. Methods and Results To test this hypothesis, we administered amoxicillin, the most commonly prescribed pediatric antibiotic, to alter gut microbiota of young, genetically hypertensive rats (study 1) and dams during gestation and lactation (study 2) and recorded their BP. Reshaping of microbiota with reductions in Firmicutes/Bacteriodetes ratio were observed. Amoxicillin treated rats had lower BP compared with untreated rats. In young rats treated with amoxicillin, the lowering effect on BP persisted even after antibiotics were discontinued. Similarly, offspring from dams treated with amoxicillin showed lower systolic BP compared with control rats. Remarkably, in all cases, a decrease in BP was associated with lowering of Veillonellaceae, which are succinate-producing bacteria. Elevated plasma succinate is reported in hypertension. Accordingly, serum succinate was measured and found lower in animals treated with amoxicillin. Conclusions Our results demonstrate a direct correlation between succinate-producing gut microbiota and early development of hypertension and indicate that reshaping gut microbiota, especially by depleting succinate-producing microbiota early in life, may have long-term benefits for hypertension-prone individuals.

Effects of different methionine levels on offspring piglets during late gestation and lactation.

Maternal dietary supplementation during gestation and lactation improves the health of piglets. The purpose of this study was to determine the effect of different levels of methionine (Met) supplementation in the sows' diet during late gestation and lactation on piglets. Thirty sows were randomly divided into three groups and fed the following diets from day 90 of gestation to day 21 of lactation: (a) control group (a basal diet containing 0.36% Met), (b) 0.48% Met group (a basal diet with additional 0.12% Met), and (c) 0.60% Met group (a basal diet with additional 0.24% Met). On day 21 after farrowing, piglets of average body weight (n = 10 per group) were selected for sample collection. The results showed that the 0.48% Met and 0.60% Met diets significantly lowered the malondialdehyde content in the piglets' serum (P < 0.05). In addition, the glutathione peroxidase content was significantly increased in the 0.48% Met group (P < 0.05) and the total glutathione content was significantly reduced in the 0.60% Met group (P < 0.05) compared to the control group. Furthermore, Met supplementation of the sows' diet was associated with alterations in 37 plasma metabolites in the piglets. In the piglets' intestinal microbiota, the relative abundances of Phascolarctobacterium and Bacteroidetes in the 0.48% Met group were higher than those in the other two groups (P < 0.05). Our results suggest that a diet including 0.48% Met during late gestation and lactation can maintain the health of piglets by increasing the antioxidant capacity and changing the intestinal microbiota composition, but a higher level of Met supplementation may increase the potential risk to piglets.

Host-microbiome interactions in human type 2 diabetes following prebiotic fibre (galacto-oligosaccharide) intake.

Aberrant microbiota composition and function have been linked to several pathologies, including type 2 diabetes. In animal models, prebiotics induce favourable changes in the intestinal microbiota, intestinal permeability (IP) and endotoxaemia, which are linked to concurrent improvement in glucose tolerance. This is the first study to investigate the link between IP, glucose tolerance and intestinal bacteria in human type 2 diabetes. In all, twenty-nine men with well-controlled type 2 diabetes were randomised to a prebiotic (galacto-oligosaccharide mixture) or placebo (maltodextrin) supplement (5·5 g/d for 12 weeks). Intestinal microbial community structure, IP, endotoxaemia, inflammatory markers and glucose tolerance were assessed at baseline and post intervention. IP was estimated by the urinary recovery of oral 51Cr-EDTA and glucose tolerance by insulin-modified intravenous glucose tolerance test. Intestinal microbial community analysis was performed by high-throughput next-generation sequencing of 16S rRNA amplicons and quantitative PCR. Prebiotic fibre supplementation had no significant effects on clinical outcomes or bacterial abundances compared with placebo; however, changes in the bacterial family Veillonellaceae correlated inversely with changes in glucose response and IL-6 levels (r -0·90, P=0·042 for both) following prebiotic intake. The absence of significant changes to the microbial community structure at a prebiotic dosage/length of supplementation shown to be effective in healthy individuals is an important finding. We propose that concurrent metformin treatment and the high heterogeneity of human type 2 diabetes may have played a significant role. The current study does not provide evidence for the role of prebiotics in the treatment of type 2 diabetes.

A case of bacteremia caused by Dialister pneumosintes and Slackia exigua in a patient with periapical abscess.

Dialister pneumosintes and Slackia exigua are both obligatory anaerobe and known to be associated with periodontal diseases and other oral infection. We report a case of blood stream infection caused by D. pneumosintes and S. exigua. This occurred in a 78-year-old female patient that presented with general weakness and fever. We revealed that she had a periapical abscess. The blood culture was positive for D. pneumosintes and S. exigua; however, identifying them was challenging. Ultimately, 16S rRNA sequencing was used to identify the organisms. The patient recovered after being treated with ceftriaxone and clindamycin. To the best of our knowledge, this is the first report of bacteremia caused by mixed infection of D. pneumosintes and S. exigua.

Interactions between oil substrates and glucose on pure cultures of ruminal lipase-producing bacteria.

The hydrolysis of free fatty acids from lipids is a prerequisite for biohydrogenation, a process that effectively saturates free fatty acids. Anaerovibrio lipolyticus 5s and Butyrivibrio fibrisolvens have long been thought to be the major contributors to ruminal lipolysis; however, Propionibacterium avidum and acnes recently have been identified as contributing lipase activity in the rumen. In order to further characterize the lipase activity of these bacterial populations, each was grown with three different lipid substrates, olive oil, corn oil, and flaxseed oil (3 %). Because different finishing rations contain varying levels of glycogen (a source of free glucose) this study also documented the effects of glucose on lipolysis. P. avidum and A. lipolyticus 5s demonstrated the most rapid rates (P < 0.05) of lipolysis for cultures grown with olive oil and flaxseed oil, respectively. A. lipolyticus, B. fibrisolvens, and P. avidum more effectively hydrolyzed flaxseed oil than olive oil or corn oil, especially in the presence of 0.02 % glucose. Conversely, P. acnes hydrolyzed corn oil more readily than olive oil or flaxseed oil and glucose had no effect on lipolytic rate. Thus, these bacterial species demonstrated different specificities for oil substrates and different sensitivities to glucose.

Antimicrobial susceptibilities and clinical sources of Dialister species.

Seventy-four strains representing the four species of the genus Dialister were isolated from various clinical samples. Dialister pneumosintes and Dialister micraerophilus were the two mainly encountered species. Fifty-five isolates were tested against 14 antimicrobial agents. Decreased susceptibilities to piperacillin, metronidazole, macrolides, fluoroquinolones, and rifampin were demonstrated. The clinical impact of these decreased susceptibilities remains to be investigated but should prompt microbiologists to perform antimicrobial susceptibility testing for clinically important Dialister spp.

Factors affecting glucose and maltose phosphorylation by the ruminal bacterium Megasphaera elsdenii.

The objectives of this study were to examine the effects of growth substrate and extracellular pH on phosphoenolpyruvate-dependent glucose phosphorylation as well as to examine how maltose is phosphorylated by the ruminal bacterium Megasphaera elsdenii B159. Phosphoenolpyruvate-dependent glucose phosphorylation by toluene-treated cells was constitutive, and glucose phosphorylation was reduced by 69% at pH 5.0. When toluene-treated cells were incubated in histidine buffer, little maltose phosphorylation occurred in the absence of inorganic phosphate. However, the addition of increasing concentrations of either potassium or sodium phosphate increased maltose phosphorylation. Maximal phosphorylation activity was observed at between 25 and 50 mM of either inorganic phosphate source. Compared with the control incubations, maltose phosphorylation was increased over threefold with 25 mM of either potassium or sodium phosphate. Phosphoglucomutase activity was detected in cell extracts of M. elsdenii B159, and this enzyme had a K(m) of 3.2 mM for glucose-1-P and a V(max) of 1836 nmol of NADP(+) reduced/mg of protein per min. Maltose was also hydrolyzed by an inducible maltase (K(m), 1.19 mM). To our knowledge, this is the first report of a maltose phosphorylase and a maltase in M. elsdenii.

Presence of lactate dehydrogenase and lactate racemase in Megasphaera elsdenii grown on glucose or lactate.

Activity of D-lactate dehydrogenase (D-LDH) was shown not only in cell extracts from Megasphaera elsdenii grown on DL-lactate, but also in cell extracts from glucose-grown cells, although glucose-grown cells contained approximately half as much D-LDH as DL-lactate-grown cells. This indicates that the D-LDH of M. elsdenii is a constitutive enzyme. However, lactate racemase (LR) activity was present in DL-lactate-grown cells, but was not detected in glucose-grown cells, suggesting that LR is induced by lactate. Acetate, propionate, and butyrate were produced similarly from both D- and L-lactate, indicating that LR can be induced by both D- and L-lactate. These results suggest that the primary reason for the inability of M. elsdenii to produce propionate from glucose is that cells fermenting glucose do not synthesize LR, which is induced by lactate.

Monensin has no effect on growth and metabolism of Megasphaera elsdenii.

A rumen strain of Megasphaera elsdenii was grown on glucose and lactate in monensin-free and monensin-supplemented medium (10 mg/L). Monensin had no effect on growth rate, growth yields, metabolic pattern and composition of cells. Growth yields of dry matter and protein were higher in cultures supplied with glucose than in cultures supplied with lactate. The bacterium compensated the lower gain of energy from fermentation of lactate by rapid utilization of this substrate. Cells grown on glucose contained more saccharide and less protein than lactate-grown cells.

[Megasphaera elsdenii--occurrence in 2,255 fecal samples of humans, chimpanzees and mice]. / Megasphaera elsdenii--über ihr Vorkommen in 2255 Stuhlproben von Menschen, Schimpansen und Mävsen.

In human gnotobiotic studies the number of cfu/g feces expresses the interspecies-specific relations within this biotop. In our study we tried to culture the resident anaerobe microflora diluting 1 g feces to 10(-6) - 10(-9), and using the cystein-blood-agar (Beerens mod. Werner) together with the dextrose-water-blue agar. M. elsdenii appear as big, smooth and yellow colonies, which may be recognised at "first sight" diagnosis. In a retrospective study of 2255 fecal samples we compared the isolation-rate of M. elsdenii in the range of greater than or equal to 10(7)/g feces from healthy people to that of persons with gastrointestinal troubles. M. elsdenii has not been isolated from fecal samples of healthy people but in 12% from those of adults and 7.4% from children, both suffering from gastrointestinal disorders. So we consider that M. elsdenii in the range of greater than or equal to 10(7)/g feces does not belong to the resident anaerobe flora of the intestine. Further studies have to establish wether its normal range may be 10(3)-10(4)/g feces like that of C. perfringens. Comparing the isolation-rate from patients with colon-carcinoma to that of patients with dermatological affections, we found a relation of 8% to 20%. The biochemical activities of our isolates were according to the Anaerobe Laboratory Manual. Additional tests e.g. API-Zym and others make us assume an activity inner the metabolism of fatty-acids and amino acids e.g. inner the aspartat and pyruvat-family.