Background:The objective of this study was to evaluate the replacement effects of milk powder(MK)and fish meal(FM)by enzymatic soybean(ESB)in diets on growth performance,immunological parameters,SCFAs production and g...Background:The objective of this study was to evaluate the replacement effects of milk powder(MK)and fish meal(FM)by enzymatic soybean(ESB)in diets on growth performance,immunological parameters,SCFAs production and gut microbiome of weaned piglets.Methods:A total of 128 piglets with initial body weight at 6.95±0.46 kg,were randomly assigned into 4 dietary treatments with 8 replicates per treatment and 4 piglets per replicate for a period of 14 d.Piglets were offered isonitrogenous and iso-energetic diets as follows:CON diet with MK and FM as high quality protein sources,ESB plus FM diet with ESB replacing MK,ESB plus MK diet with ESB replacing FM,and ESB diet with ESB replacing both MK and FM.Results:No significant differences were observed in growth performance among all treatments(P>0.05).However,piglets fed ESB plus FM or ESB diet had increased diarrhea index(P<0.01),and lower digestibility of dry matter(DM),gross energy(GE)or crude protein(CP),relative to piglets fed CON diet(P<0.01).Moreover,the inclusion of ESB in diet markedly decreased the plasma concentration of HPT and fecal concentration of butyric acid(BA)(P<0.01).The High-throughput sequencing of 16S rRNA gene V3−V4 region of gut microbiome revealed that the inclusion of ESB in diet increased the alpha diversity,and the linear discriminant analysis effect size(LEfSe)showed that piglets fed with ESB plus FM or ESB diet contained more gut pathogenic bacteria,such as g_Peptococcus,g_Veillonella and g_Helicobacter.Conclusion:The inclusion of ESB in diet did not markedly affect growth performance of piglets,but the replacement of MK or both MK and FM by ESB increased diarrhea index,which could be associated with lower nutrients digestibility and more gut pathogenic bacteria.However,piglets fed diet using ESB to replace FM did not markedly affect gut health-related parameters,indicating the potential for replacing FM with ESB in weaning diet.展开更多
Resistant starch type 3 (RS3) produced from high amylose food sources through retrogradation or enzymatic process is known to have physiological function as dietary fiber. Fermentation of RS3 by colonic microorganisms...Resistant starch type 3 (RS3) produced from high amylose food sources through retrogradation or enzymatic process is known to have physiological function as dietary fiber. Fermentation of RS3 by colonic microorganisms produced SCFA (acetate, propionate, and butyrate), maintained the health of colon, balance of gut microbiota, preventing inflammatory bowel diseases (IBD) and colon cancer. RS3 in this study was produced from IR-42 and Inpari-16 broken rice by enzymatic treatment (combination of amylase-pullulanase). The Resistant Starch was fermented for 12 and 24 h by colonic microbiota (extracted from healthy human subject), Clostiridium butyricum BCC-B2571, or Eubacterium rectale DSM 17629. SCFA produced was analyzed by gas chromatography. Treatment by amylase-pullulanase combination was advantageous to increase their RS3 content. The result showed that after enzymatic process, the RS3 content of IR-42 (41.13%) was not significantly different (p 0.05) from that of Inpari-16 (37.70%). High concentration of acetate (82.5 mM) and propionate (7.5 mM) were produced by colonic microbiota after 12 h fermentation and best concentration of butyrate (6.8 mM) was produced by colonic microbiota after 24 h fermentation. It is clear that utilization of colonic microbiota rather than single strain was better in the production of SCFA.展开更多
Objective: The objective is to explore the mechanism of inhibitory effect of three main SCFAs (acetate, propionate and butyrate) on inflammatory response of A549 cells. Methods: Human lung adenocarcinoma cells (A549 c...Objective: The objective is to explore the mechanism of inhibitory effect of three main SCFAs (acetate, propionate and butyrate) on inflammatory response of A549 cells. Methods: Human lung adenocarcinoma cells (A549 cells) were cultured, and were divided into normal control group (NC group), A. baumannii infection group (A. baumannii group), NF-κB inhibitor group (JSH group), A. baumannii infection + sodium acetate group (NaAc group), A. baumannii infection + sodium propionate group (NaPc group) and A. baumannii infection + sodium butyrate group (NaB group). Real-time quantitative PCR was used to detect the mRNA expression of NLRP3, Caspase-1, IL-1β, IL-6, and TGF-β in A549 cells. Western blotting assay was used to determine the expression of autophagy and “pyroptosis” related proteins of NRLP3, cleaved-Caspase-1 (P20), GSDMD (P30), LC-3 and Beclin-1. At the same time, the expression of NF-κB p65 protein in nucleus and cytoplasm of A549 cells was detected. The level of reactive oxygen species in A549 cells was detected by flow cytometry. Results: Compared with A. baumannii group, the mRNA expression of NLRP3, IL-1β and IL-6 in NaAc group, NaPc group and NaB group decreased significantly, the mRNA expression of Caspase-1 in NaPc group and NaB group decreased significantly, only the mRNA expression of TGF-β in NaB group increased significantly;LC3-II expression increased significantly in NaPc group and NaB group, only Beclin-1 expression increased and GSDMD (p30) expression decreased significantly in NaB group. All three kinds of SCFAs could significantly inhibit the expression of cleaved-Caspase-1 (p20) after A. baumannii infection, but there was no significant change in the protein expression of NLRP3. Compared with NC group, the production of reactive oxygen species in A. baumannii group increased significantly at 3 h after A. baumannii infection. Compared with A. baumannii group, NaB could significantly suppress the production of reactive oxygen species induced by A. baumannii. Compared with A. baumannii group, the expression of NF-κB p65 in nucleus was significantly decreased and the expression of NF-κB p65 in cytoplasm was significantly increased after 24 h pre-incubation with NaB, NaPc and NaAc, respectively. Conclusion: A. baumannii can induce inflammatory injury of pulmonary epithelial cells, and the three major SCFAs can inhibit the activation of NLRP3 inflammasome and the release of pro-inflammatory factors through NF-κB/ROS/NLRP3 pathway, which provides a new way for clinical prevention of severe inflammatory injury caused by A. baumannii infection.展开更多
Milk fat globule membrane(MFGM)possesses various nutritional and biological benefits for mammals,whereas its effects on neonatal gut microbiota and barrier integrity remained unclear.This study investigated the effect...Milk fat globule membrane(MFGM)possesses various nutritional and biological benefits for mammals,whereas its effects on neonatal gut microbiota and barrier integrity remained unclear.This study investigated the effects of MFGM administration on microbial compositions and intestinal barrier functions of neonatal piglets.Sixteen newborn piglets were randomly allocated into a CON group or MFGM group,orally administered with saline or MFGM solution(1 g/kg body weight)respectively during the first postnatal week,and all piglets were breastfed during the whole neonatal period.The present study found that the MFGM oral administration during the first postnatal week increased the plasma immunoglobulin(lg)G level,body weight and average daily gain of piglets(P<0.05)on 21 d.Addi-tionally,MFGM administration enriched fecal SCFA-producing bacteria(Ruminococaceae_UCG-002,Ruminococaceae_UCG-010,Ruminococaceae_UCG-004,Ruminococaceae_UCG-014 and[Ruminococcus]_gauvrearuii_group),SCFA concentrations(acetate,propionate and butyrate;P<0.05)and their receptor(G-protein coupled receptor 41,GPR41).Furthermore,MFGM administration promoted intestinal villus morphology(P<0.05)and barrier functions by upregulating genes of tight junctions(E-cadherin,claudin-1,occludin and zonula occludin 1[ZO-1]),mucins(mucin-13 and mucin-20)and interleukin(IL)-22(P<0.05).Positive correlation was found between the beneficial microbes and SCFA levels pairwise with the intestinal barrier genes(P<0.05).In conclusion,orally administrating MFGM during the first postnatal week stimulated SCFA-producing bacteria colonization and SCFA generation,enhanced intes-tinal barrier functions and consequently improved growth performance of neonatal piglets on 21 d.Our findings will provide new insights about MFGM intervention for microbial colonization and intestinal development of neonates during their early life.展开更多
基金This work was supported by overseas Expertise Introduction Project for Discipline Innovation(111 Project)Sichuan Agricultural University Shuangzhi plan for discipline construction project。
文摘Background:The objective of this study was to evaluate the replacement effects of milk powder(MK)and fish meal(FM)by enzymatic soybean(ESB)in diets on growth performance,immunological parameters,SCFAs production and gut microbiome of weaned piglets.Methods:A total of 128 piglets with initial body weight at 6.95±0.46 kg,were randomly assigned into 4 dietary treatments with 8 replicates per treatment and 4 piglets per replicate for a period of 14 d.Piglets were offered isonitrogenous and iso-energetic diets as follows:CON diet with MK and FM as high quality protein sources,ESB plus FM diet with ESB replacing MK,ESB plus MK diet with ESB replacing FM,and ESB diet with ESB replacing both MK and FM.Results:No significant differences were observed in growth performance among all treatments(P>0.05).However,piglets fed ESB plus FM or ESB diet had increased diarrhea index(P<0.01),and lower digestibility of dry matter(DM),gross energy(GE)or crude protein(CP),relative to piglets fed CON diet(P<0.01).Moreover,the inclusion of ESB in diet markedly decreased the plasma concentration of HPT and fecal concentration of butyric acid(BA)(P<0.01).The High-throughput sequencing of 16S rRNA gene V3−V4 region of gut microbiome revealed that the inclusion of ESB in diet increased the alpha diversity,and the linear discriminant analysis effect size(LEfSe)showed that piglets fed with ESB plus FM or ESB diet contained more gut pathogenic bacteria,such as g_Peptococcus,g_Veillonella and g_Helicobacter.Conclusion:The inclusion of ESB in diet did not markedly affect growth performance of piglets,but the replacement of MK or both MK and FM by ESB increased diarrhea index,which could be associated with lower nutrients digestibility and more gut pathogenic bacteria.However,piglets fed diet using ESB to replace FM did not markedly affect gut health-related parameters,indicating the potential for replacing FM with ESB in weaning diet.
文摘Resistant starch type 3 (RS3) produced from high amylose food sources through retrogradation or enzymatic process is known to have physiological function as dietary fiber. Fermentation of RS3 by colonic microorganisms produced SCFA (acetate, propionate, and butyrate), maintained the health of colon, balance of gut microbiota, preventing inflammatory bowel diseases (IBD) and colon cancer. RS3 in this study was produced from IR-42 and Inpari-16 broken rice by enzymatic treatment (combination of amylase-pullulanase). The Resistant Starch was fermented for 12 and 24 h by colonic microbiota (extracted from healthy human subject), Clostiridium butyricum BCC-B2571, or Eubacterium rectale DSM 17629. SCFA produced was analyzed by gas chromatography. Treatment by amylase-pullulanase combination was advantageous to increase their RS3 content. The result showed that after enzymatic process, the RS3 content of IR-42 (41.13%) was not significantly different (p 0.05) from that of Inpari-16 (37.70%). High concentration of acetate (82.5 mM) and propionate (7.5 mM) were produced by colonic microbiota after 12 h fermentation and best concentration of butyrate (6.8 mM) was produced by colonic microbiota after 24 h fermentation. It is clear that utilization of colonic microbiota rather than single strain was better in the production of SCFA.
文摘Objective: The objective is to explore the mechanism of inhibitory effect of three main SCFAs (acetate, propionate and butyrate) on inflammatory response of A549 cells. Methods: Human lung adenocarcinoma cells (A549 cells) were cultured, and were divided into normal control group (NC group), A. baumannii infection group (A. baumannii group), NF-κB inhibitor group (JSH group), A. baumannii infection + sodium acetate group (NaAc group), A. baumannii infection + sodium propionate group (NaPc group) and A. baumannii infection + sodium butyrate group (NaB group). Real-time quantitative PCR was used to detect the mRNA expression of NLRP3, Caspase-1, IL-1β, IL-6, and TGF-β in A549 cells. Western blotting assay was used to determine the expression of autophagy and “pyroptosis” related proteins of NRLP3, cleaved-Caspase-1 (P20), GSDMD (P30), LC-3 and Beclin-1. At the same time, the expression of NF-κB p65 protein in nucleus and cytoplasm of A549 cells was detected. The level of reactive oxygen species in A549 cells was detected by flow cytometry. Results: Compared with A. baumannii group, the mRNA expression of NLRP3, IL-1β and IL-6 in NaAc group, NaPc group and NaB group decreased significantly, the mRNA expression of Caspase-1 in NaPc group and NaB group decreased significantly, only the mRNA expression of TGF-β in NaB group increased significantly;LC3-II expression increased significantly in NaPc group and NaB group, only Beclin-1 expression increased and GSDMD (p30) expression decreased significantly in NaB group. All three kinds of SCFAs could significantly inhibit the expression of cleaved-Caspase-1 (p20) after A. baumannii infection, but there was no significant change in the protein expression of NLRP3. Compared with NC group, the production of reactive oxygen species in A. baumannii group increased significantly at 3 h after A. baumannii infection. Compared with A. baumannii group, NaB could significantly suppress the production of reactive oxygen species induced by A. baumannii. Compared with A. baumannii group, the expression of NF-κB p65 in nucleus was significantly decreased and the expression of NF-κB p65 in cytoplasm was significantly increased after 24 h pre-incubation with NaB, NaPc and NaAc, respectively. Conclusion: A. baumannii can induce inflammatory injury of pulmonary epithelial cells, and the three major SCFAs can inhibit the activation of NLRP3 inflammasome and the release of pro-inflammatory factors through NF-κB/ROS/NLRP3 pathway, which provides a new way for clinical prevention of severe inflammatory injury caused by A. baumannii infection.
基金the Beijing Municipal Natural Science Foundation(S170001)the National Natural Science Foundation of China(31630074,31902170,31972596 and 31902189)+2 种基金the National Key Research and Development Program of China(2016YFD0500506 and 2018YDF0501002)the Agriculture Research System of China(CARS-35)the Higher Education Discipline Innovation Project(B16044).
文摘Milk fat globule membrane(MFGM)possesses various nutritional and biological benefits for mammals,whereas its effects on neonatal gut microbiota and barrier integrity remained unclear.This study investigated the effects of MFGM administration on microbial compositions and intestinal barrier functions of neonatal piglets.Sixteen newborn piglets were randomly allocated into a CON group or MFGM group,orally administered with saline or MFGM solution(1 g/kg body weight)respectively during the first postnatal week,and all piglets were breastfed during the whole neonatal period.The present study found that the MFGM oral administration during the first postnatal week increased the plasma immunoglobulin(lg)G level,body weight and average daily gain of piglets(P<0.05)on 21 d.Addi-tionally,MFGM administration enriched fecal SCFA-producing bacteria(Ruminococaceae_UCG-002,Ruminococaceae_UCG-010,Ruminococaceae_UCG-004,Ruminococaceae_UCG-014 and[Ruminococcus]_gauvrearuii_group),SCFA concentrations(acetate,propionate and butyrate;P<0.05)and their receptor(G-protein coupled receptor 41,GPR41).Furthermore,MFGM administration promoted intestinal villus morphology(P<0.05)and barrier functions by upregulating genes of tight junctions(E-cadherin,claudin-1,occludin and zonula occludin 1[ZO-1]),mucins(mucin-13 and mucin-20)and interleukin(IL)-22(P<0.05).Positive correlation was found between the beneficial microbes and SCFA levels pairwise with the intestinal barrier genes(P<0.05).In conclusion,orally administrating MFGM during the first postnatal week stimulated SCFA-producing bacteria colonization and SCFA generation,enhanced intes-tinal barrier functions and consequently improved growth performance of neonatal piglets on 21 d.Our findings will provide new insights about MFGM intervention for microbial colonization and intestinal development of neonates during their early life.
