Effects of early postnatal gastric and colonic microbiota transplantation on piglet gut health

Background Diarrhea is a major cause of reduced growth and mortality in piglets during the suckling and weaning periods and poses a major threat to the global pig industry.Diarrhea and gut dysbiosis may in part be prevented via improved early postnatal microbial colonization of the gut.To secure better postnatal gut colonization,we hypothesized that transplantation of colonic or gastric content from healthy donors to newborn recipients would prevent diarrhea in the recipients in the post-weaning period.Our objective was to examine the impact of transplanting colonic or gastric content on health and growth parameters and paraclinical parameters in recipient single-housed piglets exposed to a weaning transition and challenged with enterotoxigenic Escherichia coli(ETEC).Methods Seventy-two 1-day-old piglets were randomized to four groups:colonic microbiota transplantation(CMT,n=18),colonic content filtrate transplantation(CcFT,n=18),gastric microbiota transplantation(GMT,n=18),or saline(CON,n=18).Inoculations were given on d 2 and 3 of life,and all piglets were milk-fed until weaning(d 20)and shortly after challenged with ETEC(d 24).We assessed growth,diarrhea prevalence,ETEC concentration,organ weight,blood parameters,small intestinal morphology and histology,gut mucosal function,and microbiota composition and diversity.Results Episodes of diarrhea were seen in all groups during both the milk-and the solid-feeding phase,possibly due to stress associated with single housing.However,CcFT showed lower diarrhea prevalence on d 27,28,and 29 compared to CON(all P<0.05).CcFT also showed a lower ETEC prevalence on d 27(P<0.05).CMT showed a higher alpha diversity and a difference in beta diversity compared to CON(P<0.05).Growth and other paraclinical endpoints were similar across groups.Conclusion In conclusion,only CcFT reduced ETEC-related post-weaning diarrhea.However,the protective effect was marginal,suggesting that higher doses,more effective modalities of administration,longer treatment periods,and better donor quality should be explored by future research to optimize the protective effects of transplantation.

Dietary protein levels and amino acid supplementation patterns alter the composition and functions of colonic microbiota in pigs

Different dietary nitrogen(N) patterns may have different effects on gut microbiota.To investigate the effects of different crude protein(CP) levels or essential amino acids(EAA) supplementation patterns on the structure and functions of colonic microbiota,42 barrows(25±0.39 kg) were randomly assigned to 7 dietary treatments including:diet 1,a high CP diet with balanced 10 EAA;diet 2,a medium CP diet with approximately 2% decreased CP level from diet 1 and balanced 10 EAA;diets 3,4,5,6 and 7,low CP diets with 4% decreased CP level from diet 1.Specifically,diet 3 was only balanced for Lys,Met,Thr and Trp;diets 4,5 and 6 were further supplemented with IIe,Val and IIe+Val on the basis of diet 3,respectively;and diet 7 was balanced for 10 EAA.Results over a 110-d trial showed that reducing the CP level by 2% or4% dramatically decreased N intake and excretion(P <0.05) in the presence of balanced 10 EAA,which was not observed when altering the EAA supplementation patterns in low CP diet(-4%).With balanced10 EAA,2% reduction in dietary CP significantly reduced Firmicutes-to-Bacteroidetes(F:B) ratio and significantly elevated the abundance of Prevotellaceae NK3 B31(P <0.05);whereas 4% reduction evidently increased the abundances of Proteobacteria,Succinivibrio and Lachnospiraceae XPB1014(P <0.05).Among the 5 low CP diets(-4%),supplementation with Ile,or Val+Ile,or balanced 10 EAA increased F:B ratio and the abundance of Proteobacteria.In addition,the predicted functions revealed that different CP levels and EAA balanced patterns dramatically altered the mRNA expression profiles of N-metabolizing genes,the "N and energy metabolism" pathways or the metabolism of some small substances,such as amino acids(AA) and vitamins.Our findings suggested that reducing the dietary CP levels by 2% to 4% with balancing 10 EAA,or only further supplementation with Ile or Val+Ile to a low protein diet(-4%) reduced the N contents entering the hindgut to various degrees,altered the abundances of N-metabolizing bacteria,and improved the abilities of N utilization.

Caecal pH is a biomarker of excessive colonic fermentation

AIM: To ascertain whether caecal pH is different in patients with irritable bowel syndrome (IBS), whose primary symptoms are bloating and distension, to healthy controls.

Impact of an oligosaccharide-based polymer on the metabolic profiles and microbial ecology of weanling pigs experimentally infected with a pathogenic E.coli

Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to carbadox.The objective of this study was to investigate the impacts of oligosaccharide-based polymer or antibiotic on the host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.Results Multivariate analysis highlighted the differences in the metabolic profiles of serum and colon digesta which were predominantly found between pigs supplemented with oligosaccharide-based polymer and antibiotic.The relative abundance of metabolic markers of immune responses and nutrient metabolisms,such as amino acids and carbohydrates,were significantly differentiated between the oligosaccharide-based polymer and antibiotic groups(q<0.2 and fold change>2.0).In addition,pigs in antibiotic had a reduced(P<0.05)relative abundance of Lachnospiraceae and Lactobacillaceae,whereas had greater(P<0.05)Clostridiaceae and Streptococcaceae in the colon digesta on d 11 post-inoculation(PI)compared with d 5 PI.Conclusions The impact of oligosaccharide-based polymer on the metabolic and microbial profiles of pigs is not fully understood,and further exploration is needed.However,current research suggest that various mechanisms are involved in the enhanced disease resistance and performance in ETEC-challenged pigs by supplementing this polymer.

Impact of humic acids on the colonic microbiome in healthy volunteers

AIMTo test the effects of humic acids on innate microbial communities of the colon.METHODSWe followed the effects of oral supplementation with humic acids (Activomin®) on concentrations and composition of colonic microbiome in 14 healthy volunteers for 45 d. 3 × 800 mg Activomin® were taken orally for 10 d followed by 3 × 400 mg for 35 d. Colonic microbiota were investigated using multicolor fluorescence in situ hybridization (FISH) of Carnoy fixated and paraffin embedded stool cylinders. Two stool samples were collected a week prior to therapy and one stool sample on days 10, 31 and 45. Forty-one FISH probes representing different bacterial groups were used.RESULTSThe sum concentration of colonic microbiota increased from 20% at day 10 to 30% by day 31 and remained stable until day 45 (32%) of humic acid supplementation (P < 0.001). The increase in the concentrations in each person was due to growth of preexisting groups. The individual microbial profile of the patients remained unchanged. Similarly, the bacterial diversity remained stable. Concentrations of 24 of the 35 substantial groups increased from 20% to 96%. Two bacterial groups detected with Bac303 (Bacteroides) and Myc657 (mycolic acid-containing Actinomycetes) FISH probes decreased (P > 0.05). The others remained unaffected. Bacterial groups with initially marginal concentrations (< 0.1 × 109/mL) demonstrated no response to humic acids. The concentrations of pioneer groups of Bifidobacteriaceae, Enterobacteriaceae and Clostridium difficile increased but the observed differences were statistically not significant.CONCLUSIONHumic acids have a profound effect on healthy colonic microbiome and may be potentially interesting substances for the development of drugs that control the innate colonic microbiome.

Mucosa-associated bacteria in two middle-aged women diagnosed with collagenous colitis

AIM:To characterize the colon microbiota in two women histologically diagnosed with collagenous colitis using a culture-independent method.METHODS:Biopsies were taken from the ascending colon and the total DNA was extracted.Universal bacterial primers were used to amplify the bacterial 16S rRNA genes.The amplicons were then cloned into competent Escherichia coli cells.The clones were sequenced and identified by comparison to known sequences.RESULTS:The clones could be divided into 44 different phylotypes.The microbiota was dominated by Firmicutes and Bacteroidetes.Seven phylotypes werefound in both patients and constituted 47.5% of the total number of clones.Of these,the most dominating were clones similar to Bacteroides cellulosilyticus,Bacteroides caccae,Bacteroides thetaiotaomicron,Bacteroides uniformis and Bacteroides dorei within Bacteroidetes.Sequences similar to Faecalibacterium prausnitzii and Clostridium citroniae were also found in both patients.CONCLUSION:A predominance of potentially pathogenic Bacteroides spp.,and the presence of clones showing similarity to Clostridium clostridioforme were found but the overall colon microbiota showed similarities to a healthy one.Etiologies for collagenous colitis other than an adverse bacterial flora must also be considered.

Yogurt-derived Lactobacillus plantarum Q16 alleviated high-fat diet-induced non-alcoholic fatty liver disease in mice

Accumulating evidence revealed that some probiotics regulated lipid metabolism and alleviated diet-induced non-alcoholic fatty liver disease(NAFLD).This study mainly explored whether yogurt-derived Lactobacillus plantarum Q16 modulated lipid and energy metabolism,and suppressed microbial dysbiosis in high-fat diet(HFD)-fed mice.Results showed that oral administration of L.plantarum Q16 improved serum and hepatic lipid profile.Protein analysis showed that L.plantarum Q16 could reduce hepatic lipid content by reducing the expression of FAS,ACC,SCD-1,Srebp-lc and ATGL,but increasing expression levels of CPT-1α,PPAR-αand ATGL.Meanwhile,L.plantarum Q16 also improved hepatic energy metabolism by regulating FGF21/adiponectin/AMPKα/PGC-1αsignaling pathway.Metagenomic analysis also discovered that L.plantarum Q16 increased species diversity and richness of intestinal microbiota,promoted proliferation of beneficial commensals and suppressed the growth of endotoxin-producing microorganisms in the colon of HFD-fed mice.Overall,L.plantarum Q16 protected against HFD-induced NAFLD by improving hepatic profile and regulating colonic microbiota composition.

Porcine intestinal antimicrobial peptide as an in-feed antibiotic alternative improves intestinal digestion and immunity by shaping the gut microbiota in weaned piglets

Antibiotic resistance of pathogens,which is caused by the abuse of in-feed antibiotics,threatens the sustainable development of livestock production.The present study aimed to investigate the efficiency of porcine intestinal antimicrobial peptide(PIAP)as an alternative to in-feed antibiotics in terms of growth performance,intestinal morphology,digestive enzymes and immunity,and microbiota community of the post-weaning piglets.A total of 204 piglets(Duroc×Landrace×Yorkshire,weaned at 28 d age)with a similar body weight of 7.97±1.04 kg were randomly allocated to 4 groups(51 piglets per group):(1)control group:basal diet;(2)AB group:antibiotic,basal diet+chlortetracycline(1000 mg/kg from d 1 to24;500 mg/kg from d 25 to 37);(3)P1 group:basal diet+a relatively low dose of PIAP(400 mg/kg from d 1 to 24;300 mg/kg from d 25 to 37);(4)P2 group,basal diet+a relatively high dose of PIAP(600 mg/kg from d 1 to 24;500 mg/kg from d 25 to 37).The results showed that serum indicators of hepatocyte damage and relative organ weight were not affected by these treatments(P>0.05).Compared with the AB treatment,the P1 treatment remarkably decreased jejunal crypt depth and increased jejunal and ileal villus height:crypt depth ratio(P<0.05).The values of jejunal maltase,lactase,sucrase,intestinal alkaline phosphatase,and secretory immunoglobulin A(SIg A)in the P1 group were sharply increased compared with those in the control and P2 groups(P<0.05).Compared with the control group,the P1 group decreased serum concentrations of D-lactate,diamine oxidase,and endotoxin(P<0.05),and increased the abundance of Lactobacillus reuteri(P<0.05)in the colonic feces.Furthermore,there was a positive correlation between the abundance of L.reuteri and the concentrations of maltase,lactase,sucrase,and SIg A(P<0.05).Collectively,dietary supplementation with a relatively low dose of PIAP(400 mg/kg from d 1 to 24;300 mg/kg from d 25 to 37)demonstrates beneficial effects on intestinal morphology,digestive enzymes,immunity,and permeability by shaping the gut microbiota composition in weaned piglets.This study will provide a valuable reference for using PIAP as an in-feed antibiotic alternative in swine production.

Supplementation of a lacto-fermented rapeseed-seaweed blend promotes gut microbial-and gut immune-modulation in weaner piglets

Background:The direct use of medical zinc oxide in feed will be abandoned after 2022 in Europe,leaving an urgent need for substitutes to prevent post-weaning disorders.Results:This study investigated the effect of using rapeseed-seaweed blend(rapeseed meal added two brown macroalgae species Ascophylum nodosum and Saccharina latissima)fermented by lactobacilli(FRS)as feed ingredients in piglet weaning.From d 28 of life to d 85,the piglets were fed one of three different feeding regimens(n=230 each)with inclusion of 0%,2.5% and 5% FRS.In this period,no significant difference of piglet performance was found among the three groups.From a subset of piglets(n=10 from each treatment),blood samples for hematology,biochemistry and immunoglobulin analysis,colon digesta for microbiome analysis,and jejunum and colon tissues for histopathological analyses were collected.The piglets fed with 2.5% FRS manifested alleviated intraepithelial and stromal lymphocytes infiltration in the gut,enhanced colon mucosa barrier relative to the 0% FRS group.The colon microbiota composition was determined using V3 and V1-V8 region 16S rRNA gene amplicon sequencing by Illumina NextSeq and Oxford Nanopore MinION,respectively.The two amplicon sequencing strategies showed high consistency between the detected bacteria.Both sequencing strategies indicated that inclusion of FRS reshaped the colon microbiome of weaned piglets with increased Shannon diversity.Prevotella stercorea was verified by both methods to be more abundant in the piglets supplied with FRS feed,and its abundance was positively correlated with colonic mucosa thickness but negatively correlated with blood concentrations of leucocytes and IgG.Conclusions:FRS supplementation relieved the gut lymphocyte infiltration of the weaned piglets,improved the colon mucosa barrier with altered microbiota composition.Increasing the dietary inclusion of FRS from 2.5% to 5% did not lead to further improvements.

Trace amounts of antibiotic altered metabolomic and microbial profiles of weaned pigs infected with a pathogenic E.coli

Background:Our previous study has shown that supplementation of trace amounts of antibiotic exacerbated the detrimental effects of enterotoxigenic E.coli(ETEC)infection and delayed the recovery of pigs that may be associated with modified metabolites and metabolic pathways.Therefore,the objective of this study was to explore the impacts of trace levels of antibiotic(carbadox)on host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.Results:The multivariate analysis highlighted a distinct metabolomic profile of serum and colon digesta between trace amounts of antibiotic(TRA;0.5 mg/kg carbadox)and label-recommended dose antibiotic(REC;50 mg/kg carbadox)on d 5 post-inoculation(PI).The relative abundance of metabolomic markers of amino acids,carbohydrates,and purine metabolism were significantly differentiated between the TRA and REC groups(q<0.2).In addition,pigs in REC group had the highest(P<0.05)relative abundance of Lactobacillaceae and tended to have increased(P<0.10)relative abundance of Lachnospiraceae in the colon digesta on d 5 PI.On d 11 PI,pigs in REC had greater(P<0.05)relative abundance of Clostridiaceae compared with other groups,whereas had reduced(P<0.05)relative abundance of Prevotellaceae than pigs in control group.Conclusions:Trace amounts of antibiotic resulted in differential metabolites and metabolic pathways that may be associated with its slow responses against ETEC F18 infection.The altered gut microbiota profiles by labelrecommended dose antibiotic may contribute to the promotion of disease resistance in weaned pigs.

Perioperative changes in lower gut microbiocenosis in patients with colon and rectum cancer

This clinical study was designed to determinate the value of colon microbiota microflora in colorectal cancer patients.We investigate pre-and post-operation stool samples in patients with different tumor localizations,trying to correlate those to postoperative complications.Our findings shows that if opportunistic pathogenic microflora levels are elevated before operations,it can lead to further microbiota disbalance in postoperative period,especially in cases of antibacterial prophylaxis or treatment.Further,this changes increase a chance of receiving some complications after surgery.So,we suggest an analysis of stool microbiota in patients with colorectal cancer to be performed prior and after the operation.Furthermore,aperioperative decontamination therapy should be performed in cases of pathogenic microflora levels elevation.Still,monitoring aperioperative microbiota changes is not a routine in most recommendations and we assume that including this relatively unencumbered patient testing in preoperative survey will help to reduce a number of postoperative complications.