Background： Colibacillosis caused by enterotoxigenic Escherichia coil （E. coil} results in economic losses in the poultry industry. Antibiotics are usually used to control colibacillosis, however, E. coli has varyin...Background： Colibacillosis caused by enterotoxigenic Escherichia coil （E. coil} results in economic losses in the poultry industry. Antibiotics are usually used to control colibacillosis, however, E. coli has varying degrees of resistance to different antibiotics. Therefore the use of probiotics is becoming accepted as an alternative to antibiotics. In this study, we evaluated the effects of Clostfidium butyricum （C. butyficum） on growth performance, immune response, intestinal barrier function, and digestive enzyme activity in broiler chickens challenged with Eschefichia coli （E. coil） K88. Methods： The chickens were randomly divided into four treatment groups for 28 days. Negative control treatment （NC） consisted of birds fed a basal diet without E. coil K88 challenge and positive control treatment （PC） consisted of birds fed a basal diet and challenged with E. coil K88. C. buO/ricum probiotic treatment （CB） consisted of birds fed a diet containing 2 x 107 cfu C. buO/ricum/kg of diet and challenged with E. coil K88. Colistin sulfate antibiotic treatment （CS） consisted of birds fed a diet containing 20 mg colistin sulfate/kg of diet and challenged with E. coil K88. Results： The body weight （BW） and average day gain （ADG） in the broilers of CB group were higher （P 〈 0.05） than the broilers in the PC group overall except the ADG in the 14-21 d post-challenge. The birds in CB treatment had higher （P 〈 0.05） concentration of tumor necrosis factor-a （TNF-a） at 3 and 7 d post-challenge, and higher （P 〈 0.05） concentration of interleukin-4 （IL-4） at 14 d post-challenge than those in the PC treatment group. The concentration of serum endotoxin in CB birds was lower （P 〈 0.05） at 21 d post-challenge, and the concentrations of serum diamine oxidase in CB birds were lower （P 〈 0.05） at 14 and 21 d post-challenge than in PC birds. Birds in CB treatment group had higher （P 〈 0.05） jejunum villi height than those in PC, NC, or CS treatment at 展开更多
Background:Enterotoxigenic Escherichia coli(ETEC)K88 commonly colonize in the small intestine and keep releasing enterotoxins to impair the intestinal barrier function and trigger inflammatory reaction.Although Lactob...Background:Enterotoxigenic Escherichia coli(ETEC)K88 commonly colonize in the small intestine and keep releasing enterotoxins to impair the intestinal barrier function and trigger inflammatory reaction.Although Lactobacillus salivarius(L.salivarius)has been reported to enhance intestinal health,it remains to be seen whether there is a functional role of L.salivarius in intestinal inflammatory response in intestinal porcine epithelial cell line(IPEC-J2)when stimulated with ETEC K88.In the present study,IPEC-J2 cells were first treated with L.salivarius followed by the stimulation of ETEC K88 for distinct time period.ETEC K88 adherent status,pattern recognition receptors(PRRs)mRNA,mitogen-activated protein kinase(MAPK)and nuclear factor-κB(NF-κB)activation,the release of pro-inflammation cytokines and cell integrity were examined.Results:Aside from an inhibited adhesion of ETEC K88 to IPEC-J2 cells,L.salivarius was capable of remarkably attenuating the expression levels of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-8,Toll-like receptor(TLR)4,nucleotide-binding oligomerization domain(NOD)-like receptor pyrin domain-containing protein(NLRP)3 and NLRP6.This alternation was accompanied by a significantly decreased phosphorylation of p38 MAPK and p65 NF-κB during ETEC K88 infection with L.salivarius pretreatment.Western blot analysis revealed that L.salivarius increased the expression levels of zona occludens 1(ZO-1)and occludin(P<0.05)in ETEC K88-infected IPEC-J2 cells.Compared with ETEC K88-infected groups,the addition of L.salivarius as well as extra inhibitors for MAPKs and NF-κB to ETEC K88-infected IPEC-J2 cells had the capability to reduce pro-inflammatory cytokines.Conclusions:Collectively,our results suggest that L.salivarius might reduce inflammation-related cytokines through attenuating phosphorylation of p38 MAPK and blocking the NF-κB signaling pathways.Besides,L.salivarius displayed a potency in the enhancement of IPEC-J2 cell integrity.展开更多
Casein glycomacropeptide(CGMP) is a bioactive peptide derived from milk with multiple functions. This study was aimed at evaluating the effects of CGMP as a potential feed additive on growth performance,intestinal mor...Casein glycomacropeptide(CGMP) is a bioactive peptide derived from milk with multiple functions. This study was aimed at evaluating the effects of CGMP as a potential feed additive on growth performance,intestinal morphology, intestinal barrier permeability and inflammatory responses of Escherichia coli K88(E. coli K88) challenged piglets. Eighteen weaning piglets were randomly assigned to three groups.Control group and K88 challenged group received a basal diet, and CGMP treated group received the basal diet supplemented with 1% of CGMP powder. The trail lasted for 12 days, K88 was orally administered to the piglets of K88 challenged group and CGMP treated group on days 8-10. The results showed that the diet containing 1% CGMP significantly alleviated the decrease in average daily gain(P <0.05),increase in pathogenic bacteria amounts in intestinal contents(P < 0.05), intestinal morphology(P > 0.05) and barrier permeability damage(P < 0.05), and acute inflammatory response(P < 0.05)induced by E. coli K88 infection. In conclusion, CGMP supplementation in the diet protected the weaning piglets against E. coli K88 infection.展开更多
The adsorption properties of Cu 2＋ -loaded montmorillonite clays （MMT-Cu） for Escherichia coli K 88 as a function of time,bacteria concentrations,pH,ionic strength and temperature were investigated.The results show...The adsorption properties of Cu 2＋ -loaded montmorillonite clays （MMT-Cu） for Escherichia coli K 88 as a function of time,bacteria concentrations,pH,ionic strength and temperature were investigated.The results showed that the bacteria adsorption onto MMT-Cu surface reached equilibrium after 90 min.The percentages of E.coli K 88 adsorbed onto the surfaces of MMT-Cu and montmorillonite clays （MMT） at equilibrium were 88.9% and 56.5%,respectively.Scanning electron microscopy revealed that a lot of E.coli K 88 adhered to the surface of MMT-Cu.The zeta potential of MMT-Cu was relatively high as compared to that of MMT.The adsorptive ability of MMT-Cu for E.coli K 88 was higher than that of MMT （P 0.05）.Moreover,pH,ionic strength and temperature produced a strong influence on the extent of E.coli K 88 adsorption to surface of MMT-Cu and MMT.The mechanism of adsorption of E.coli onto MMT-Cu may involve electrostatic attraction and physiochemical properties of bacterial cell walls and minerals surfaces.展开更多
Background:This study aimed to investigate the effects of oral administration of Enterococcus faecium NCIMB10415(E.faecium)on intestinal development,immunological parameters and gut microbiota of neonatal piglets chal...Background:This study aimed to investigate the effects of oral administration of Enterococcus faecium NCIMB10415(E.faecium)on intestinal development,immunological parameters and gut microbiota of neonatal piglets challenged with enterotoxigenic Escherichia coli K88(ETEC).A total of 961-day-old sow-reared piglets were randomly assigned to 2 groups,with 48 piglets in each group.The piglets were from 16 litters(6 piglets each litter),and 3 piglets each litter were allocated to the E.faecium-supplemented(PRO)group,while the other 3 piglets were allocated to the control(CON)group.After colostrum intake,piglets in the PRO group were orally administrated with 3×10~9 CFU E.faecium per day for a period of one week.On day 8,one piglet per litter from each group was challenged(CON+ETEC,PRO+ETEC)or not(CON-ETEC,PRO-ETEC)with ETEC in a 2×2 factorial arrangement of treatments.On day 10(2 days after challenge),blood and tissue samples were obtained from piglets.Results:Before ETEC challenge,there were no significant differences for the average daily gain(ADG)and fecal score between the two groups of piglets.After ETEC challenge,the challenged piglets had greater fecal score compared to the non-challenged piglets,whereas E.faecium administration was able to decrease the fecal score.Piglets challenged with ETEC had shorter villous height,deeper crypt depth,and reduced number of goblet cells in the jejunum and decreased m RNA abundance of claudin-1 in the ileum,whereas increased the percentage of lymphocytes,concentrations of IL-1βin the plasma and TNF-αin the ileal mucosa,as well as increased the m RNA abundances of innate immunity-related genes in the ileum tissue.These deleterious effects caused by ETEC were partly alleviated by feeding E.faecium.In addition,piglets in PRO-ETEC group had decreased the percentage of CD8^+T cells of the peripheral blood when compared to those in CON-ETEC group.Moreover,E.faecium administration increased Verrucomicrobia at phylum level and decreased Bilophila at genus level.Conclusions:These r展开更多
Background:Necroptosis and pyroptosis are newly identified forms of programmed cell death,which play a vital role in development of many gastrointestinal disorders.Although plant polyphenols have been reported to prot...Background:Necroptosis and pyroptosis are newly identified forms of programmed cell death,which play a vital role in development of many gastrointestinal disorders.Although plant polyphenols have been reported to protect intestinal health,it is still unclear whether there is a beneficial role of plant polyphenols in modulating necroptosis and pyroptosis in intestinal porcine epithelial cell line(IPEC-1)infected with enterotoxigenic Escherichia coli(ETEC)K88.This research was conducted to explore whether plant polyphenols including protocatechuic acid(PCA)and quercetin(Que),attenuated inflammation and injury of IPEC-1 caused by ETEC K88 through regulating necroptosis and pyroptosis signaling pathways.Methods:IPEC-1 cells were treated with PCA(40μmol/L)or Que(10μmol/L)in the presence or absence of ETEC K88.Results:PCA and Que decreased ETEC K88 adhesion and endotoxin level(P<0.05)in cell supernatant.PCA and Que increased cell number(P<0.001)and decreased lactate dehydrogenases(LDH)activity(P<0.05)in cell supernatant after ETEC infection.PCA and Que improved transepithelial electrical resistance(TEER)(P<0.001)and reduced fluorescein isothiocyanate-labeled dextran(FD4)flux(P<0.001),and enhanced membrane protein abundance of occludin,claudin-1 and ZO-1(P<0.05),and rescued distribution of these tight junction proteins(P<0.05)after ETEC infection.PCA and Que also declined cell necrosis ratio(P<0.05).PCA and Que reduced mRNA abundance and concentration of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-8(P<0.001),and down-regulated gene expression of toll-like receptors 4(TLR4)and its downstream signals(P<0.001)after ETEC infection.PCA and Que down-regulated protein abundance of total receptor interacting protein kinase 1(t-RIP1),phosphorylated-RIP1(p-RIP1),p-RIP1/t-RIP1,t-RIP3,p-RIP3,mixed lineage kinase domain-like protein(MLKL),p-MLKL,dynamin-related protein 1(DRP1),phosphoglycerate mutase 5(PGAM5)and high mobility group box 1(HMGB1)(P<0.05)after ETEC infection.Moreover,PCA and Que reduced protein abundanc展开更多
基金supported by the International Cooperation Project of Zhejiang Province(No.2012C14031)Innovative Research Team Program of Zhejiang Province(No.2011R50025)
文摘Background： Colibacillosis caused by enterotoxigenic Escherichia coil （E. coil} results in economic losses in the poultry industry. Antibiotics are usually used to control colibacillosis, however, E. coli has varying degrees of resistance to different antibiotics. Therefore the use of probiotics is becoming accepted as an alternative to antibiotics. In this study, we evaluated the effects of Clostfidium butyricum （C. butyficum） on growth performance, immune response, intestinal barrier function, and digestive enzyme activity in broiler chickens challenged with Eschefichia coli （E. coil） K88. Methods： The chickens were randomly divided into four treatment groups for 28 days. Negative control treatment （NC） consisted of birds fed a basal diet without E. coil K88 challenge and positive control treatment （PC） consisted of birds fed a basal diet and challenged with E. coil K88. C. buO/ricum probiotic treatment （CB） consisted of birds fed a diet containing 2 x 107 cfu C. buO/ricum/kg of diet and challenged with E. coil K88. Colistin sulfate antibiotic treatment （CS） consisted of birds fed a diet containing 20 mg colistin sulfate/kg of diet and challenged with E. coil K88. Results： The body weight （BW） and average day gain （ADG） in the broilers of CB group were higher （P 〈 0.05） than the broilers in the PC group overall except the ADG in the 14-21 d post-challenge. The birds in CB treatment had higher （P 〈 0.05） concentration of tumor necrosis factor-a （TNF-a） at 3 and 7 d post-challenge, and higher （P 〈 0.05） concentration of interleukin-4 （IL-4） at 14 d post-challenge than those in the PC treatment group. The concentration of serum endotoxin in CB birds was lower （P 〈 0.05） at 21 d post-challenge, and the concentrations of serum diamine oxidase in CB birds were lower （P 〈 0.05） at 14 and 21 d post-challenge than in PC birds. Birds in CB treatment group had higher （P 〈 0.05） jejunum villi height than those in PC, NC, or CS treatment at
基金funded by the Tianjin Natural Science Foundation(18JCYBJC30000)the National Natural Science Foundation of China(31702147)the Tianjin“131”Innovative Talents Team(20180338).
文摘Background:Enterotoxigenic Escherichia coli(ETEC)K88 commonly colonize in the small intestine and keep releasing enterotoxins to impair the intestinal barrier function and trigger inflammatory reaction.Although Lactobacillus salivarius(L.salivarius)has been reported to enhance intestinal health,it remains to be seen whether there is a functional role of L.salivarius in intestinal inflammatory response in intestinal porcine epithelial cell line(IPEC-J2)when stimulated with ETEC K88.In the present study,IPEC-J2 cells were first treated with L.salivarius followed by the stimulation of ETEC K88 for distinct time period.ETEC K88 adherent status,pattern recognition receptors(PRRs)mRNA,mitogen-activated protein kinase(MAPK)and nuclear factor-κB(NF-κB)activation,the release of pro-inflammation cytokines and cell integrity were examined.Results:Aside from an inhibited adhesion of ETEC K88 to IPEC-J2 cells,L.salivarius was capable of remarkably attenuating the expression levels of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-8,Toll-like receptor(TLR)4,nucleotide-binding oligomerization domain(NOD)-like receptor pyrin domain-containing protein(NLRP)3 and NLRP6.This alternation was accompanied by a significantly decreased phosphorylation of p38 MAPK and p65 NF-κB during ETEC K88 infection with L.salivarius pretreatment.Western blot analysis revealed that L.salivarius increased the expression levels of zona occludens 1(ZO-1)and occludin(P<0.05)in ETEC K88-infected IPEC-J2 cells.Compared with ETEC K88-infected groups,the addition of L.salivarius as well as extra inhibitors for MAPKs and NF-κB to ETEC K88-infected IPEC-J2 cells had the capability to reduce pro-inflammatory cytokines.Conclusions:Collectively,our results suggest that L.salivarius might reduce inflammation-related cytokines through attenuating phosphorylation of p38 MAPK and blocking the NF-κB signaling pathways.Besides,L.salivarius displayed a potency in the enhancement of IPEC-J2 cell integrity.
基金supported by the earmarked fund for Modern Agro-industry Technology Research System(CARS-36)
文摘Casein glycomacropeptide(CGMP) is a bioactive peptide derived from milk with multiple functions. This study was aimed at evaluating the effects of CGMP as a potential feed additive on growth performance,intestinal morphology, intestinal barrier permeability and inflammatory responses of Escherichia coli K88(E. coli K88) challenged piglets. Eighteen weaning piglets were randomly assigned to three groups.Control group and K88 challenged group received a basal diet, and CGMP treated group received the basal diet supplemented with 1% of CGMP powder. The trail lasted for 12 days, K88 was orally administered to the piglets of K88 challenged group and CGMP treated group on days 8-10. The results showed that the diet containing 1% CGMP significantly alleviated the decrease in average daily gain(P <0.05),increase in pathogenic bacteria amounts in intestinal contents(P < 0.05), intestinal morphology(P > 0.05) and barrier permeability damage(P < 0.05), and acute inflammatory response(P < 0.05)induced by E. coli K88 infection. In conclusion, CGMP supplementation in the diet protected the weaning piglets against E. coli K88 infection.
基金supported by the National Natural Science Foundation of China (No.30471255)the Program for New Century Excellent Talents in University(No.NCET-06-0913)
文摘The adsorption properties of Cu 2＋ -loaded montmorillonite clays （MMT-Cu） for Escherichia coli K 88 as a function of time,bacteria concentrations,pH,ionic strength and temperature were investigated.The results showed that the bacteria adsorption onto MMT-Cu surface reached equilibrium after 90 min.The percentages of E.coli K 88 adsorbed onto the surfaces of MMT-Cu and montmorillonite clays （MMT） at equilibrium were 88.9% and 56.5%,respectively.Scanning electron microscopy revealed that a lot of E.coli K 88 adhered to the surface of MMT-Cu.The zeta potential of MMT-Cu was relatively high as compared to that of MMT.The adsorptive ability of MMT-Cu for E.coli K 88 was higher than that of MMT （P 0.05）.Moreover,pH,ionic strength and temperature produced a strong influence on the extent of E.coli K 88 adsorption to surface of MMT-Cu and MMT.The mechanism of adsorption of E.coli onto MMT-Cu may involve electrostatic attraction and physiochemical properties of bacterial cell walls and minerals surfaces.
基金supported by the Projects of The National Key Research and Development Program of China(grant number 2016YFD0501204)Sichuan provincial project on S&T application and demonstration(grant number2016CC0070)the project on commercialization of research findings under funding of government of Sichuan province(grant number16ZHSF0385).
文摘Background:This study aimed to investigate the effects of oral administration of Enterococcus faecium NCIMB10415(E.faecium)on intestinal development,immunological parameters and gut microbiota of neonatal piglets challenged with enterotoxigenic Escherichia coli K88(ETEC).A total of 961-day-old sow-reared piglets were randomly assigned to 2 groups,with 48 piglets in each group.The piglets were from 16 litters(6 piglets each litter),and 3 piglets each litter were allocated to the E.faecium-supplemented(PRO)group,while the other 3 piglets were allocated to the control(CON)group.After colostrum intake,piglets in the PRO group were orally administrated with 3×10~9 CFU E.faecium per day for a period of one week.On day 8,one piglet per litter from each group was challenged(CON+ETEC,PRO+ETEC)or not(CON-ETEC,PRO-ETEC)with ETEC in a 2×2 factorial arrangement of treatments.On day 10(2 days after challenge),blood and tissue samples were obtained from piglets.Results:Before ETEC challenge,there were no significant differences for the average daily gain(ADG)and fecal score between the two groups of piglets.After ETEC challenge,the challenged piglets had greater fecal score compared to the non-challenged piglets,whereas E.faecium administration was able to decrease the fecal score.Piglets challenged with ETEC had shorter villous height,deeper crypt depth,and reduced number of goblet cells in the jejunum and decreased m RNA abundance of claudin-1 in the ileum,whereas increased the percentage of lymphocytes,concentrations of IL-1βin the plasma and TNF-αin the ileal mucosa,as well as increased the m RNA abundances of innate immunity-related genes in the ileum tissue.These deleterious effects caused by ETEC were partly alleviated by feeding E.faecium.In addition,piglets in PRO-ETEC group had decreased the percentage of CD8^+T cells of the peripheral blood when compared to those in CON-ETEC group.Moreover,E.faecium administration increased Verrucomicrobia at phylum level and decreased Bilophila at genus level.Conclusions:These r
基金provided by National Key R&D Program of China(2022YFD1300403)National Natural Science Foundation of China(No.U22A20517,32272906,and 31802070)Wuhan Science and Technology Bureau(No.2022020801010391)。
文摘Background:Necroptosis and pyroptosis are newly identified forms of programmed cell death,which play a vital role in development of many gastrointestinal disorders.Although plant polyphenols have been reported to protect intestinal health,it is still unclear whether there is a beneficial role of plant polyphenols in modulating necroptosis and pyroptosis in intestinal porcine epithelial cell line(IPEC-1)infected with enterotoxigenic Escherichia coli(ETEC)K88.This research was conducted to explore whether plant polyphenols including protocatechuic acid(PCA)and quercetin(Que),attenuated inflammation and injury of IPEC-1 caused by ETEC K88 through regulating necroptosis and pyroptosis signaling pathways.Methods:IPEC-1 cells were treated with PCA(40μmol/L)or Que(10μmol/L)in the presence or absence of ETEC K88.Results:PCA and Que decreased ETEC K88 adhesion and endotoxin level(P<0.05)in cell supernatant.PCA and Que increased cell number(P<0.001)and decreased lactate dehydrogenases(LDH)activity(P<0.05)in cell supernatant after ETEC infection.PCA and Que improved transepithelial electrical resistance(TEER)(P<0.001)and reduced fluorescein isothiocyanate-labeled dextran(FD4)flux(P<0.001),and enhanced membrane protein abundance of occludin,claudin-1 and ZO-1(P<0.05),and rescued distribution of these tight junction proteins(P<0.05)after ETEC infection.PCA and Que also declined cell necrosis ratio(P<0.05).PCA and Que reduced mRNA abundance and concentration of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-8(P<0.001),and down-regulated gene expression of toll-like receptors 4(TLR4)and its downstream signals(P<0.001)after ETEC infection.PCA and Que down-regulated protein abundance of total receptor interacting protein kinase 1(t-RIP1),phosphorylated-RIP1(p-RIP1),p-RIP1/t-RIP1,t-RIP3,p-RIP3,mixed lineage kinase domain-like protein(MLKL),p-MLKL,dynamin-related protein 1(DRP1),phosphoglycerate mutase 5(PGAM5)and high mobility group box 1(HMGB1)(P<0.05)after ETEC infection.Moreover,PCA and Que reduced protein abundanc
