Clostridium Cluster Research Articles

The association between gut butyrate-producing bacteria and non-small-cell lung cancer.

BackgroundRecently, it has been found that the gut microbiota may affect the development of lung cancer through the “gut‐lung axis.” To investigate this relationship, we performed this study to determine whether the gut microbiota in non‐small‐cell lung cancer (NSCLC) patients is different from that in healthy adults.MethodsQuantitative PCR (qPCR) was used to detect the expression levels of eight gut butyrate‐producing bacteria in healthy adults and NSCLC patients. We enrolled 30 patients with NSCLC and 30 subjects from 100 healthy adults after matching for age and sex.ResultsCompared to healthy adults, most of the gut butyrate‐producing bacteria in NSCLC patients were significantly decreased; these included Faecalibacterium prausnitzii, Clostridium leptum, Clostridial cluster I, Ruminococcus spp., Clostridial Cluster XIVa, and Roseburia spp. Among the gut butyrate‐producing bacteria, we analyzed Clostridial cluster IV and Eubacterium rectale were not decreased in NSCLC patients.ConclusionsWe conclude that NSCLC patients had gut butyrate‐producing bacteria dysbiosis. Further studies should be performed to investigate the underlying mechanisms of how these specific bacteria affect lung cancer progression and prognosis.

α-Linolenic Acid-Rich Diet Influences Microbiota Composition and Villus Morphology of the Mouse Small Intestine.

α-Linolenic acid (ALA) is well-known for its anti-inflammatory activity. In contrast, the influence of an ALA-rich diet on intestinal microbiota composition and its impact on small intestine morphology are not fully understood. In the current study, we kept adult C57BL/6J mice for 4 weeks on an ALA-rich or control diet. Characterization of the microbial composition of the small intestine revealed that the ALA diet was associated with an enrichment in Prevotella and Parabacteroides. In contrast, taxa belonging to the Firmicutes phylum, including Lactobacillus, Clostridium cluster XIVa, Lachnospiraceae and Streptococcus, had significantly lower abundance compared to control diet. Metagenome prediction indicated an enrichment in functional pathways such as bacterial secretion system in the ALA group, whereas the two-component system and ALA metabolism pathways were downregulated. We also observed increased levels of ALA and its metabolites eicosapentanoic and docosahexanoic acid, but reduced levels of arachidonic acid in the intestinal tissue of ALA-fed mice. Furthermore, intestinal morphology in the ALA group was characterized by elongated villus structures with increased counts of epithelial cells and reduced epithelial proliferation rate. Interestingly, the ALA diet reduced relative goblet and Paneth cell counts. Of note, high-fat Western-type diet feeding resulted in a comparable adaptation of the small intestine. Collectively, our study demonstrates the impact of ALA on the gut microbiome and reveals the nutritional regulation of gut morphology.

Hermetia illucens larvae as a Fishmeal replacement alters intestinal specific bacterial populations and immune homeostasis in weanling piglets

Hermetia illucens larvae meal (HILM) are rich in proteins and chitin, and represent an innovative feed ingredient for animals. However, little is known about the intestinal bacteria and immune homeostasis response of HILM as a fishmeal replacement on weanling piglets. Thus, this study aimed to investigate the changes in specific ileal and cecal bacterial populations and their metabolic profiles, and ileal immune indexes in weanling piglets fed with a diet containing HILM. A total of 128 weanling piglets were fed either a basal diet or 1 of 3 diets with 1%, 2%, and 4% HILM (HI0, HI1, HI2, and HI4, respectively). Each group consisted of 8 pens (replicates), with 4 pigs per pen. After 28 d of feeding, 8 barrows per treatment were euthanized, the ileal and cecal digesta, and ileal mucosa were collected for analyzing bacterial population and metabolic profiles, and immune indexes, respectively. Results showed that HILM increased (P < 0.05, maximum in HI2) the number of Lactobacillus and Bifidobacterium in the ileum and cecum, but quadratically decreased (P < 0.05, minimum in HI2) the number of Escherichia coli. In the cecum, the number of Firmicutes, Ruminococcus, Clostridium cluster IV, and Prevotella showed a quadratic response to increasing (P < 0.05, maximum in HI2) HILM levels. Lactate and butyrate concentrations in the ileum and cecum were quadratically increased (P < 0.05, maximum in HI2) with increasing HILM levels. In the cecum, the amines, phenol, and indole compounds concentrations were quadratically decreased (P < 0.05, minimum in HI2) with increasing HILM levels, while total short-chain fatty acids and acetate concentrations were quadratically increased (P < 0.05, maximum in HI2). In the ileum, the TLR4, NF-κB, MyD88, and TNF-α mRNA expressions were quadratically decreased (P < 0.05, minimum in HI2) with increasing HILM levels, while the mRNA expression of IL-10, barrier function (MUC1, ZO-1, Occludin, and Claudin-2), and development-related genes (IGF-1, GLP-2, and EGF) was quadratically increased (P < 0.05, maximum in HI2). Furthermore, the changes in the mucosal gene expression were associated with changes in the bacterial populations and their metabolites. Collectively, these results showed that a diet supplemented with 2% HILM affected specific bacterial populations and metabolic profiles, and maintained ileal immune status. These findings provide new insights into the use of insect meal as a suitable alternative protein source for swine feeding.

Effects of an alginate-containing variable-viscosity enteral nutrition formula on defecation, intestinal microbiota, and short-chain fatty acid production

Abstract This study aimed to elucidate the effects of a liquid polymeric formula containing alginate, a source of water- soluble dietary fiber, on stool form, intestinal microbiota, and the production of short-chain fatty acid production. Participants were 11 elderly patients with malnutrition and poor oral intake, who required enteral feeding. The alginate-containing liquid formula, which characteristically changes state from liquid to semi-solidified depending on pH, was administered for 4 weeks. This variable-viscosity liquid formula significantly improved the values of nutritional indices and stool form. Analysis of fecal specimens revealed significant decreases in pH. The concentrations of short-chain fatty acids did not change significantly in stool samples, but acetic acid and propionic acid were significantly increased in blood samples. The proportion of Clostridium cluster XI in fecal microbiota was also significantly increased. The alginate-containing variable-viscosity liquid formula significantly improved stool form, lowered fecal pH, and short-chain fatty acid concentrations in blood.

A randomized controlled trial investigating the effect of a diet low in fermentable oligosaccharides, disaccharides, monosaccharides, and polyols on the intestinal microbiome and inflammation in patients with ulcerative colitis: study protocol for a randomized controlled trial

BackgroundNo conclusive treatment is available for irritable bowel disease (IBD). Adherence to a diet low in fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAPs) might alleviate clinical symptoms of IBD. However, no study has investigated the effect of low FODMAPs diet on the intestinal microbiota and inflammatory biomarkers in patients with IBD. The aim of current study is to examine the effect a low FODMAP diet on IBD symptoms, inflammation, and the intestinal microbiota in patients with ulcerative colitis.Methods and analysisThis study is a randomized clinical trial. Thirty patients with mild to moderate ulcerative colitis will be randomly allocated to receive a low FODMAP diet (n = 15) or to continue their usual diet as control (n = 15), for 4 weeks. The quantity of intestinal microbiota including Clostridium cluster IV, Faecalibacterium prausnitzii, Rosburia spp., Lactobacillus spp., Bifidobacteria spp., Akkermansia muciniphila, Bacteroides fragilis, and Ruminococcus spp., and the Firmicutes to Bacteroidetes ratio and calprotectin and lactoferrin levels will be explored in fecal samples from patients. In addition, anthropometric measures and biochemical assessments including serum concentrations of highly sensitive-C reactive protein (hs-CRP), tumour necrosis factor-α (TNF-α) and IL-1β will be taken from patients at baseline and end of the study. The study has been registered in IRCT (IRCT20181126041763N1; registration date: 2019-01-18).DiscussionConsumption of a low-FODMAP diet might decrease systemic and intestinal inflammation, change the bacterial population in the gut, and modulate clinical symptoms in patients with ulcerative colitis. Further studies investigating the effect of such a diet on other variables, including other bacterial species and inflammatory cytokines, are required to confirm future findings of this trial.

DNA damage in colon mucosa of Pirc rats, an Apc-driven model of colon tumorigenesis

APC mutation is the first event triggering colon carcinogenesis (CRC). The contribution of APC to colon mucosa DNA damage is not well characterized yet. Similarly, the role of genotoxin-producer gut microorganisms is unclear. DNA strand breaks and oxidative damage were measured in Pirc rats, mutated in Apc, with the COMET assay at age 1 (T1) and 11 months (T11), i.e. in absence and presence of colon adenomas. In Pirc colon mucosa a 2-fold increase in the mean level of DNA oxidative damage was found at T11 compared to T1. Moreover, the analysis of DNA damage distribution showed that the proportion of Pirc mucosa cells in the highest DNA damage class was increased compared to wt rats at T1 and T11 months (p < 0.05 and <0.001, respectively). The analysis of colon mucosa-associated microbiota composition showed that this result was not attributable to the presence of genotoxin-producer bacteria B. fragilis nor E. coli. However, Pirc colon mucosa was enriched in Clostridium cluster XI, harmful bacteria in the large intestine, while the wt colon mucosa was enriched in Clostridium cluster IV. This work provides an original way to investigate the interplay between Apc and gut microbiota in affecting DNA stability during CRC.

Hop-derived prenylflavonoid isoxanthohumol suppresses insulin resistance by changing the intestinal microbiota and suppressing chronic inflammation in high fat diet-fed mice.

To assess whether the hop-derived polyphenol isoxanthohumol suppresses insulin resistance by changing the intestinal microbiota. Male C57BL/6J mice (7 weeks of age) were divided into five groups (n = 9-10): Normal Diet (ND), High Fat Diet (HFD), HFD + low dose isoxanthohumol (0.01%IX), HFD + medium dose isoxanthohumol (0.03% IX), and HFD + high dose isoxanthohumol (0.1% IX). Oral glucose tolerance tests (OGTTs) were performed at 4 and 8 weeks, and insulin tolerance tests (ITTs) were performed at 13 weeks. 16S rRNA gene sequencing analyses revealed the fecal microbiota profiles, and the relative abundance of Akkermansia muciniphila and Clostridium cluster XI was calculated by qRT-PCR. Plasma lipopolysaccharide (LPS) levels were measured by ELISA, and mRNA expression levels of tumor necrosis factor (TNF)-α, and interleukin (IL)-1β in epididymal adipose tissues were measured by qRT-PCR. Isoxanthohumol showed antibacterial activity towards several bacterial species and mitigated impaired glucose tolerance and insulin resistance induced by the HFD in a dose-dependent manner, as shown by OGTTs and ITTs. The concentration of phylum Verrucomicrobia bacteria dramatically increased in the 0.1% IX group, the relative abundance of A. muciniphila increased, and that of Clostridium cluster XI decreased. Moreover, the intake of isoxanthohumol decreased the levels of plasma LPS and mRNA expression of TNF-α and IL-1β in epididymal adipose tissues. We found that isoxanthohumol can suppress HFD-induced insulin resistance by changing the intestinal microbiota and reducing the expression of inflammation factors.

Effects of Antibiotic Pretreatment of an Ulcerative Colitis-Derived Fecal Microbial Community on the Integration of Therapeutic Bacteria In Vitro.

Fecal microbiota transplantation (FMT) is a proposedly useful strategy for the treatment of gastrointestinal (GI) disorders through remediation of the patient gut microbiota. However, its therapeutic success has been variable, necessitating research to uncover mechanisms that improve patient response. Antibiotic pretreatment has been proposed as one method to enhance the success rate by increasing niche availability for introduced species. Several limitations hinder exploring this hypothesis in clinical studies, such as deleterious side effects and the development of antimicrobial resistance in patients. Thus, the purpose of this study was to evaluate the use of an in vitro, bioreactor-based, colonic ecosystem model as a form of preclinical testing by determining how pretreatment with the antibiotic rifaximin influenced engraftment of bacterial strains sourced from a healthy donor into an ulcerative colitis-derived defined microbial community. Distinct species integrated under the pretreated and untreated conditions, with the relative rifaximin resistance of the microbial strains being an important influencer. However, both conditions resulted in the integration of taxa from Clostridium clusters IV and XIVa, a concomitant reduction of Proteobacteria, and similar decreases in metabolites associated with poor health status. Our results agree with the findings of similar research in the clinic by others, which observed no difference in primary patient outcomes whether or not patients were given rifaximin prior to FMT. We therefore conclude that our model is useful for screening for antibiotics that could improve efficacy of FMT when used as a pretreatment.IMPORTANCE Patients with gastrointestinal disorders often exhibit derangements in their gut microbiota, which can exacerbate their symptoms. Replenishing these ecosystems with beneficial bacteria through fecal microbiota transplantation is thus a proposedly useful therapeutic; however, clinical success has varied, necessitating research into strategies to improve outcomes. Antibiotic pretreatment has been suggested as one such approach, but concerns over harmful side effects have hindered testing this hypothesis clinically. Here, we evaluate the use of bioreactors supporting defined microbial communities derived from human fecal samples as models of the colonic microbiota in determining the effectiveness of antibiotic pretreatment. We found that relative antimicrobial resistance was a key determinant of successful microbial engraftment with rifaximin (broad-spectrum antibiotic) pretreatment, despite careful timing of the application of the therapeutic agents, resulting in distinct species profiles from those of the control but with similar overall outcomes. Our model had results comparable to the clinical findings and thus can be used to screen for useful antibiotics.

Effect of amount of milk replacer fed and the processing of corn in starter on growth performance, nutrient digestibility, and rumen and fecal fibrolytic bacteria of dairy calves

The aim of this study was to evaluate effects of milk replacer (MR) feeding rate and processing of corn in calf starter (CS) on growth performance, nutrient digestibility, and rumen and fecal fibrolytic bacteria in dairy calves. Holstein male calves (n = 48, 2-3 d of age) were randomly assigned to 1 of 4 treatments with a 2 × 2 factorial arrangement of MR level of 0.749 kg of MR/d (LO) or up to 1.498 kg of MR/d (HI); and whole corn or flaked corn in textured CS. Calves were weaned by reducing MR offered by 50% during wk 6. Intakes of MR and CS were recorded daily, whereas body weight (BW) was measured weekly. Rumen fluid and fecal matter were collected at wk 5 and 8 to quantify fibrolytic bacteria and nutrient digestibility. Data were analyzed as a completely randomized design using mixed model ANOVA. Repeated measures were used as appropriate. Calves fed HI had greater average daily gain than calves fed LO at wk 2, 3, 4, and 5, yet at wk 7 calves fed HI had lower average daily gain compared with calves fed LO. Starter intake was greater for calves fed LO compared with HI at wk 4, 5, 6, and 7. During wk 5 and 8, calves fed LO had increased ADF and NDF digestibility compared with calves fed HI. During wk 5, dry matter and organic matter digestibility were lower for LO-fed calves compared with HI-fed calves, but during wk 8 the opposite was observed, with HI-fed calves having lower dry matter and organic matter digestibility than LO-fed calves. At wk 5, Clostridium cluster IV and Butyrivibrio fibrisolvens proportions in rumen fluid tended to be higher and Clostridium cluster IV, Fecalibacterium sp., and Prevotella sp. proportions in fecal matter were higher in calves fed LO compared with HI. From wk 8 to 16, dry matter intake was unaffected by treatment; however, energy efficiency was greater in calves fed LO, causing LO calves to have higher BW gain during this period. Greater starter digestibility was observed for calves fed LO versus HI in concert with increased fibrolytic bacteria proportions (wk 5) in fecal and rumen samples, which resulted in greater postweaning BW gain and similar BW and frame measurements by 16 wk of age. Overall the results show that rate of MR feeding has a larger effect than the processing of corn in CS on performance, fiber digestibility, and rumen and fecal fibrolytic bacterial communities.

Worm expulsion is independent of alterations in composition of the colonic bacteria that occur during experimental Hymenolepis diminuta-infection in mice

ABSTRACT The tapeworm Hymenolepis diminuta fails to establish in mice. Given the potential for helminth-bacteria interaction in the gut and the influence that commensal bacteria exert on host immunity, we tested if worm expulsion was related to alterations in the gut microbiota. Specific pathogen-free (SPF) mice, treated with broad-spectrum antibiotics, or germ-free wild-type mice were infected with H. diminuta, gut bacterial composition assessed by 16S rRNA gene sequencing, and worm counts, blood eosinophilia, goblet cells, splenic IL-4, -5 and -10, and colonic cytokines/chemokines mRNA were assessed. Effects of a PBS-soluble extract of adult H. diminuta on bacterial growth in vitro was tested. H. diminuta-infected mice displayed increased α and β diversity in colonic mucosa-associated and fecal bacterial communities, characterized by increased Lachnospiraceae and clostridium cluster XIVa. In vitro analysis revealed that the worm extract promoted the growth of anaerobic bacteria on M2GSC agar. H. diminuta-infection was accompanied by increased Th2 immune responses, and colon from infected mice had increased levels of IL-10, IL-25, Muc2, trefoil factor 3, and β2-defensin mRNA. SPF-mice treated with antibiotics, or germ-free mice, expelled H. diminuta with kinetics similar to control SPF mice. In both settings, measurements of Th2-immune responses were not significantly different across the groups. Thus, while infection with H. diminuta results in subtle but distinct changes to the colonic microbiota, we have no evidence to support an essential role for gut bacteria in the expulsion of the worm from the mouse host.

In Vitro Evaluation of the Effects of Tylosin on the Composition and Metabolism of Canine Fecal Microbiota

Simple SummaryThe antibiotic-responsive enteropathy is a common canine chronic disorder for which tylosin represents an effective widely used therapeutic option, although its mechanism of action, beyond the well-known antibacterial activity, is still unclear. Given the beneficial role of prebiotic substrates for gut health, positive outcomes deriving from the association of tylosin with some prebiotic oligosaccharides might be supposed. The present study investigated in vitro the effects of tylosin, alone or supplemented with fructooligosaccharides, galactooligosaccharides, or xylooligosaccharides, on the composition and activity of the fecal microbiota of healthy dogs. It was partially confirmed that the antibacterial effect of tylosin, given the reduction of some microbial populations and metabolites, e.g., volatile fatty acids. Interestingly, the association of tylosin with prebiotics revealed counteracting effects on some undesirable changes exerted by tylosin, e.g., the reduction of bacteria generally considered beneficial such as lactobacilli and Clostridium cluster XIVa as well as volatile fatty acids, i.e., microbial fermentative end-products that are recognized as essential for enterocytes homeostasis.The present study investigated the in vitro effects of tylosin (TYL), alone or associated with prebiotics (PRE), on selected canine fecal parameters. Eight treatments were set up: control diet with no addition of substrates; TYL; Fructooligosaccharides (FOS); Galactooligosaccharides (GOS); Xylooligosaccharides (XOS); TYL + FOS; TYL + GOS; TYL + XOS. The flasks (five for treatment), containing a canine fecal suspension (prepared with the feces of healthy adult dogs) and the residue of an in vitro digested dry dog food, were incubated in an anaerobic chamber at 39 °C. TYL and PRE were added at a concentration of 0.2 and 1 g/L, respectively. Samples were collected after 6 and 24 h for analyses. PRE decreased pH values, iso-butyrate, and iso-valerate throughout the incubation; increased lactobacilli, cadaverine, and, tendentiously, total volatile fatty acids after 6 h; increased n-butyrate, putrescine, spermidine, and reduced spermine and E. coli after 24 h. TYL resulted in lower total volatile fatty acids and lactobacilli and higher Clostridium cluster I after 6 h and higher pH values, spermidine, and E. coli throughout the study. When associated with TYL, PRE counteracted some undesirable effects of the antibiotic such as the decrease of lactobacilli and Clostridium cluster XIVa at both 6 and 24 h. In the present study, TYL exhibited inhibitory effects on canine fecal microbiota partially counteracted by PRE supplementation.

Effect of early antibiotic intervention on specific bacterial communities and immune parameters in the small intestine of growing pigs fed different protein level diets

Antibiotics are designed to affect gut microbiota and subsequently gut homeostasis. However, limited information exists about short- and long-term effects of early antibiotic intervention (EAI) on gut homeostasis (especially for the small intestine) of pigs following antibiotic withdrawal. We investigated the impact of EAI on specific bacterial communities, microbial metabolites and mucosal immune parameters in the small intestine of later-growth-stage pigs fed with diets differing in CP levels. Eighteen litters of piglets were fed creep feed with or without antibiotics from day 7 to day 42. At day 42, pigs within each group were offered a normal- or low-CP diet. Five pigs per group were slaughtered at days 77 and 120. At day 77, EAI increased Enterobacteriaceae counts in the jejunum and ileum and decreased Bifidobacterium counts in the jejunum and ileum (P < 0.05). Moreover, tryptamine, putrescine, secretory immunoglobulin (Ig) A and IgG concentrations in the ileum and interleukin-10 (IL-10) mRNA and protein levels in the jejunum and ileum were decreased in pigs with EAI (P < 0.05). At day 120, EAI only suppressed Clostridium cluster XIVa counts in the jejunum and ileum (P < 0.05). These results suggest that EAI has a short-term effect on specific bacterial communities, amino acid decarboxylation and mucosal immune parameters in the small intestine (particularly in the ileum). At days 77 and 120, feeding a low-CP diet affected Bifidobacterium, Clostridium cluster IV, Clostridium cluster XIVa and Enterobacteriaceae counts in the jejunum or ileum (P < 0.05). Moreover, feeding a low-CP diet increased the concentrations of Igs in the jejunum and decreased pro-inflammatory cytokines levels in the jejunum and ileum (P < 0.05). At day 120, feeding a low-CP diet increased short-chain fatty acid concentrations, reduced ammonia and spermidine concentrations and up-regulated genes related to barrier function in the jejunum and ileum (P < 0.05). These results suggest that feeding a low-CP diet changes specific bacterial communities and intestinal metabolite concentrations and modifies mucosal immune parameters. These findings contribute to our understanding on the duration of the impact of EAI on gut homeostasis and may provide basis data for nutritional modification in young pigs after antibiotic treatment.

Health-promoting effects of Lactobacillus-fermented barley in weaned pigs challenged with Escherichia coli K88 +

Fermented feeds are being considered as practical alternatives to antimicrobial growth promoters (AGP) supplemented in nursery pig diets. This study aimed to investigate health-promoting effects of fermented barley in weaned pigs challenged with Escherichia coli K88 +. A total of 37 piglets were weaned at 21 ± 1 day of age (6.41 ± 0.47 kg of BW) and assigned to either of the following five treatment groups: (1) unchallenged control (UCC; n = 7), (2) challenged control (CC; n = 7), (3) AGP (CC + 0.1% AGP; n = 7), (4) Ferm1 (challenged and fed homofermentative Lactobacillus plantarum (Homo)-fermented barley; n = 8) and (5) Ferm2 (challenged and fed heterofermentative L. buchneri (Hetero)-fermented barley; n = 8). The control diet included unfermented barley. Barley was fermented with either Homo or Hetero for 90 days under anaerobic conditions. On day 10, all pigs except those in UCC group were orally inoculated with E. coli K88 + (6 × 109 colony forming units/ml). The pre-planned orthogonal test was performed to compare (1) UCC and CC, (2) CC and AGP, (3) CC and Ferm1 + Ferm2, as well as (4) Ferm1 and Ferm2. Challenged control pigs showed shorter (P < 0.05) villus height (VH) in the duodenum and deeper (P < 0.05) crypt depth (CD) in the jejunum than UCC pigs. The AGP group had higher (P < 0.05) VH and lower (P < 0.05) IL-6 gene expression in the jejunum compared with CC group. Compared to CC, Ferm1 and Ferm2 had decreased (P < 0.05) CD in the duodenum, IL-6 gene expression in the jejunum and rectal temperature at 24 h post-challenge. Pigs fed fermented barley diets showed greater (P < 0.05) faecal abundance of Clostridium Cluster IV and Lactobacilli than those fed UCC diet. Ferm2-fed pigs showed lower (P < 0.05) concentrations of band cells, eosinophils and lymphocytes at 6, 24 and 48 h after challenge, respectively, and lower (P < 0.05) faecal abundance of Enterobacteriaceae 24 h after challenge than the Ferm1-fed pigs. In conclusion, the substitution of unfermented barley with fermented barley in a nursery diet showed similar results as those shown by AGP supplementation in terms of enhancing the intestinal morphology and modulating faecal microbiota composition, as well as down-regulating the pro-inflammatory cytokines; therefore, fermented barley can be a possible nutritional strategy for managing nursery pigs fed diets without in-feed AGP.

The effects of a nutritional supplement containing salacinol in neonatal Thoroughbred foals.

ABSTRACTA nutritional supplement containing salacinol (NSS) was administered to Thoroughbred foals daily beginning 21 days after birth, and clinical signs and intestinal microbiota were analyzed. The average number of days for which foals exhibited a fever between 21 and 110 days after birth was determined. The number of days was significantly reduced, by approximately 1/3, in the NSS group compared with the control group. Furthermore, improved weight gain was observed in the NSS group compared with the control group. By analyzing the intestinal microbiota, it was determined that the ratio of Clostridium cluster XIVa increased after 3 weeks of NSS administration. These results demonstrate that the daily administration of NSS might improve the intestinal environment of neonatal foals and be useful for health.

132 Short-term and long-term intake of high-level pea fiber specifically affects the bacterial community and metabolites in the cecum of pigs

Abstract Current study was conducted to investigate the influence of short-term and long-term feeding of high-level pea-hull fiber (PF) on the bacterial community and metabolites in the cecum of pigs. A total of 50 healthy weaned Duroc × Landrace × Yorkshire piglets (7.2 ± 0.5 kg) were selected and randomly allocated in two groups with 5 replicates (5 piglets per replicate) in each group. Pigs in control group were given basal diets. For pigs in another group, expanded corn in the basal diet was replaced by PF at 10%, 20% or 30% for the post-weaning period (from weaning to 30 d post-weaning), growing period (30–90 d post-weaning), and finishing period (90–160 d post-weaning), respectively. At the end of the first and the last period, one pig from each replicate was sacrificed and the cecal digesta was collected immediately. Results showed that: 1) short-term or long-term intake of high-level PF had no significant effect on the ADFI and ADG of the pigs (P &amp;gt; 0.05), but the F/G was significantly decreased in the long-term fed pigs (P &amp;lt; 0.05). 2) The long-term feeding of PF significantly increased the concentration of cecal volatile fatty acids and decreased the ratio of propionate (P &amp;lt; 0.05). 3) Compared with control group, the ratio of phylum Firmicutes in the cecum of piglets with short-term feeding of PF was reduced by 5.6%, with an increase of Bacteroides-Prevotella-Porphyromonas (BPP), Enterococcus, Clostridium cluster IV, Bacteroidetes, Lactobacillus and Desulfovibrio desulfuricans (P &amp;lt; 0.05). While in the cecum of pigs with long-term feeding of PF, the ratio of phylum Bacteroidetes was increased by 4.8%, with an increase of D. desulfuricans (P &amp;lt; 0.01) and a decrease of BPP, Helicobacter-Flexispira-Wollinella, Streptococcus, Enterococcus and Clostridium cluster I (P &amp;lt; 0.05). Results indicate that bacteria in the hindgut of pigs can rapidly response to the high-level PF in the diet.

Effects of gastrointestinal digested polyphenolic enriched extracts of Chilean currants (Ribes magellanicum and Ribes punctatum) on in vitro fecal microbiota.

Chilean currants (Ribes magellanicum and Ribes punctatum) are wild polyphenol-rich berries with interesting bioactivities in several in vitro models. The aim of this study was to investigate the effects of the pre-digested PEE (polyphenol-enriched extract) in a simulated colon model. Fruits were extracted, submitted to simulated gastrointestinal digestion and further colonic fermentation with feces from healthy human donors. Samples were taken at 1, 4, 8 and 24h of incubation, monitoring pH, ammonia, branched-chain fatty acids (BCFA), short-chain fatty acids (SCFA) and bacterial growth. FOS (fructooligosaccharides) and fecal slurry without treatments were positive and negative control, respectively. Both Ribes species reduced (p<0.05) both BCFA and SCFA at 24h. R. punctatum promoted the growth (p<0.05) of beneficial bacteria such as Clostridium cluster XIVa, and Faecalibacterium prausnitzii; while a trend to increase Akkermansia muciniphila was observed. R. magellanicum increased (p<0.05) Clostridium cluster XIVa population. Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. remained unaffected. Our results suggest that polyphenols from R. punctatum and R. magellanicum may modulate both bacterial metabolism and some selected gut beneficial bacteria under simulated conditions. Therefore, Chilean currants might be useful as supplements to maintain a healthy colon; however, further in vivo studies are needed to confirm their effect and their mechanisms.

Characterization of the Cultivable Microbiota in Fresh and Stored Mature Human Breast Milk.

Besides nutritional components, breast milk contains diverse microbes, which may be involved in colonization of the infant gut. Expressed milk is often stored for few days in the refrigerator. The aim of this study was to determine the abundance, prevalence and diversity of facultative and strict anaerobic bacteria using culture-dependent and -independent methods, and to determine changes in milk microbial and chemical composition during storage. Samples of mature breast milk from 21 women were collected 3–6 months post-partum and were analyzed fresh or after anaerobic storage for 6 days at 4°C. The cultivable bacterial population was analyzed using the most probable number (MPN) method or plate counts and different media. The abundance of major bacterial groups was determined using quantitative PCR and 16S rRNA gene sequencing. Lactose, lactate, short chain fatty acids (SCFA) and human milk oligosaccharides (HMO) were analyzed using chromatography techniques. Highest mean viable cell counts were obtained in yeast casitone fatty acids (YCFA) broth supplied with mucin (log 4.2 ± 1.8 cells/ml) and lactose (log 3.9 ± 1.4 cells/ml), or Columbia broth (log 3.0 ± 0.7 cells/ml). Mean total bacterial counts estimated by qPCR was 5.3 ± 0.6 log cells/ml, with Firmicutes being the most abundant phylum. The most prevalent bacterial groups were Streptococcus spp. (15/19 samples), Enterobacteriaceae (13/19) and Lactobacillus/Lactococcus/Pediococcus group (12/19). While the average total number of bacterial cells did not change significantly during storage, the prevalence of strict anaerobic Bacteroidetes increased threefold, from 3/19 to 9/19, while in 7 samples Clostridium clusters IV or XIVa became detectable after storage. Major HMO were not degraded. Lactate was present in 18/21 samples after storage (2.3–18.0 mM). Butyrate was detected in 15/21 and 18/21 samples before and after storage, respectively, at concentrations ranging from 2.5 to 5.7 mM. We demonstrate enhanced prevalence and/or abundance of viable strict anaerobes from the Bacteroidetes and Clostridiales after 6-day anaerobic storage of human milk. Our data indicate that anaerobic cold storage did not markedly change total viable bacterial load, while HMO profiles were stable. Anaerobic cold storage of human milk for up to 6 days may be suitable for preserving milk quality for potential microbial transfer to the infant gut.

Comparison between a commercial blend of functional oils and monensin on the performance and microbiota of coccidiosis-challenged broilers

The aim of the study was to evaluate the effects of a cashew nut shell oil and commercial castor oil blend (CNSL-Castor oil) on the performance and microbiota of broiler chickens with and without coccidiosis challenge. A total of 864 one-day-old male chicks (Cobb) were randomly distributed to receive 6 treatments (8 pens/treatment; 18 chicks/pen) in a 3 × 2 factorial, with 3 additives (control [non-additives], 100ppm sodium monensin, or 0.15% CNSL-Castor oil blend), and 2 levels of coccidiosis challenge at 14 D of age (unchallenged or inoculated by gavage with 1mL of solution containing oocysts sporulated with Eimeria tenella, Eimeria acervulina, and Eimeria maxima). No differences in productive performance were observed among treatments in the pre-challenge period and in unchallenged birds (P > 0.05). Seven-days post-challenge, birds receiving monensin performed better than birds in the positive control group (non-additive and challenge) or in the CNSL-Castor oil group (P > 0.05). However, 14 D post-challenge, birds supplemented with CNSL-Castor oil presented higher weight gain and better feed conversion (P > 0.05), without any change in feed intake (P > 0.05). During the accumulated period (1 to 42 D of age), the live weight, weight gain, and feed intake did not differ between the CNSL-Castor oil and monensin groups, both of which presented higher values than the positive control. Lactobacillus spp. and Clostridium perfringens numbers were increased in the challenged birds (P < 0.05). CNSL-Castor oil supplementation reduced Clostridium cluster XIV, C. perfringens, and S. aureus, compared with the monensin and control groups (P > 0.05). In addition, the CNSL-Castor oil group presented the highest number of Lactobacillus spp. copies, followed by the monensin and positive control groups (P > 0.05). Thus, monensin and CNSL-Castor oil effectively minimized the impact of coccidiosis at different times. While monensin acts as an antimicrobial, CNSL-Castor oil modulates the intestinal microbiota with antimicrobial action against gram-positive bacteria, mainly C. perfringens and S. aureus.

Changes in cecum microbial community in response to total sulfur amino acid (TSAA: DL-methionine) in antibiotic-free and supplemented poultry birds

The effect of essential total sulfur amino acids (TSAA) like methionine and cysteine on the cecal microbiome of broilers was investigated at 2 different time points (days 21 and 42) of broiler rearing. A total of 360-day-old Cobb male broiler chicks were randomly distributed to 6 dietary treatments in a 2 × 3 factorial arrangement, with 2 levels of antibiotic growth promoters (AGP: 0 and 0.05%) and 3 levels of TSAA (DL-methionine) either for starter (0.7, 0.8, and 0.9%) or finisher chicks (0.52, 0.62, and 0.72%), labeled as diets 1 to 6. Cecal digesta from each replicate (n = 10) were sampled on days 21 and 42. DNA was extracted for the amplification of the V4 region of bacterial 16S rRNA genes and subjected to Illumina sequencing. Bioinformatic analyses were performed using QIIME, Mothur, and ad hoc tools and functional profiles of the inferred metagenome were analyzed using PICRUST. Statistical difference was determined by 2-way ANOVA and PERMANOVA. Clustering of cecal communities using PCoA showed clear separation of microbial communities based on age (P < 0.05) of birds and between low and medium/ high levels of TSAA (DL-methionine). At day 21, bacterial richness and diversity were higher than at day 42 where Clostridium cluster XI and Lactobacillus were found most abundant. No variability in taxonomic richness at the genus level was observed with AGP and DL-methionine supplementation. Interbird variation for richness was greater at day 42 compared to day 21. The mean fold difference of richness was greater (1.5 mean fold) with diets 1 and 6, suggesting interactive effects of AGP and TSAA (DL-methionine) in the diet. KEGG function profiles calculated by PICRUST suggest that the cecal microbiome increased glycolysis and energy generation correlated with increased dietary TSAA (DL-methionine) supplementation levels during the late broiler growth period (day 42). This study increases our knowledge of microbial dynamics and functions that are relevant to host nutrition and performance that may help us tailoring alternative strategies for raising poultry birds under antibiotic-free conditions.

Gut Microbiota in Bipolar Depression and Its Relationship to Brain Function: An Advanced Exploration.

The mechanism of bipolar disorder is unclear. Growing evidence indicates that gut microbiota plays a pivotal role in mental disorders. This study aimed to find out changes in the gut microbiota in bipolar depression (BD) subjects following treatment with quetiapine and evaluate their correlations with the brain and immune function. Totally 36 subjects with BD and 27 healthy controls (HCs) were recruited. The severity of depression was evaluated with the Montgomery-Asberg depression rating scale (MADRS). At baseline, fecal samples were collected and analyzed by quantitative polymerase chain reaction (qPCR). T lymphocyte subsets were measured to examine immune function. Near-infrared spectroscopy (NIRS) was used to assess brain function. All BD subjects received quetiapine treatment (300 mg/d) for four weeks, following which the fecal microbiota and immune profiles were reexamined. Here, we first put forward the new concept of brain-gut coefficient of balance (B-GCB), which referred to the ratio of [oxygenated hemoglobin]/(Bifidobacteria to Enterobacteriaceae ratio), to analyze the linkage between the gut microbiota and brain function. At baseline, the CD3+ T cell proportion was positively correlated with log10 Enterobacter spp count, whereas the correlativity between the other bacteria and immune profiles were negative. Log10 B-GCB was positively correlated with CD3+ T cell proportion. In subjects with BD, counts of Faecalibacterium prausnitzii, Bacteroides–Prevotella group, Atopobium Cluster, Enterobacter spp, and Clostridium Cluster IV were higher, whereas the log10 (B/E) were lower than HCs (B/E refers to Bifidobacteria to Enterobacteriaceae ratio and represents microbial colonization resistance). After treatment, MADRS scores were reduced, whereas the levels of Eubacterium rectale, Bifidobacteria, and B/E increased. The composition of the gut microbiota and its relationship to brain function were altered in BD subjects. Quetiapine treatment was effective for depression and influenced the composition of gut microbiota in patients. Clinical Trial Registration: http://www.chictr.org.cn/index.aspx, identifier ChiCTR-COC-17011401, URL: http://www.chictr.org.cn/listbycreater.aspx.