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
Helminth-induced Th2 immunity and gut microbiota have been recently shown to be highly effective in modulating metabolic syndromes in animal models.This study aimed to determine whether maternal immunity and microbial...Helminth-induced Th2 immunity and gut microbiota have been recently shown to be highly effective in modulating metabolic syndromes in animal models.This study aimed to determine whether maternal immunity and microbial factors affect the induction and development of obesity in offspring.Here,Heligomosomoides polygyrus(Hp)-infected or control female C57BL/6J mice mated with normal males and their offspring were fed a high-fat diet(HFD)for 9 weeks after weaning.Our results showed that Hp-induced maternal outcomes during gestation and lactation significantly impacted offspring metabolic phenotypes.This was evidenced by results showing that offspring from helminth-infected mothers on an HFD(Hp-offspring+HFD)gained significantly less body weight than those from uninfected mothers(Cont-offspring+HFD).Hp-offspring+HFD exhibited no Th2 phenotype but displayed a pattern of gut microbiota composition similar to that of Hp-infected mothers.Cross-fostering experiments confirmed that the helminth-induced maternal attenuation of offspring obesity was mediated through both prenatal and postnatal effects.Our results further showed that helminth-infected dams and their offspring had a markedly altered gut microbiome composition,with increased production of short-chain fatty acids(SCFAs).Intriguingly,Hp-infected mothers and Hp-offspring+HFD showed increased SCFA receptor(GPR)expression in adipose and colonic tissues compared to noninfected mothers and Cont-offspring+HFD,respectively.Moreover,SCFA supplementation to the pups of uninfected control mothers during lactation protected against HFD-induced weight gain,which corresponded with changes in gut bacterial colonization.Collectively,our findings provide new insights into the complex interaction of maternal immune status and gut microbiome,Hp infection,and the immunity and gut microbiome in obese-prone offspring in infant life.展开更多
Many marine bacteria are difcult to culture because they are dormant,rare or found in low-abundances.Enrichment culturing has been widely tested as an important strategy to isolate rare or dormant microbes.However,man...Many marine bacteria are difcult to culture because they are dormant,rare or found in low-abundances.Enrichment culturing has been widely tested as an important strategy to isolate rare or dormant microbes.However,many more mechanisms remain uncertain.Here,based on 16S rRNA gene high-throughput sequencing and metabolomics technology,it was found that the short-chain fatty acids(SCFAs)in metabolites were signifcantly correlated with uncultured bacterial groups during enrichment cultures.A pure culture analysis showed that the addition of SCFAs to media also resulted in high efciency for the isolation of uncultured strains from marine sediments.As a result,238 strains belonging to 10 phyla,26 families and 82 species were successfully isolated.Some uncultured rare taxa within Chlorobi and Kiritimatiellaeota were successfully cultured.Amongst the newly isolated uncultured microbes,most genomes,e.g.bacteria,possess SCFA oxidative degradation genes,and these features might aid these microbes in better adapting to the culture media.A further resuscitation analysis of a viable but non-culturable(VBNC)Marinilabiliales strain verifed that the addition of SCFAs could break the dormancy of Marinilabiliales in 5 days,and the growth curve test showed that the SCFAs could shorten the lag phase and increase the growth rate.Overall,this study provides new insights into SCFAs,which were frst studied as resuscitation factors in uncultured marine bacteria.Thus,this study can help improve the utilisation and excavation of marine microbial resources,especially for the most-wanted or key players.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grants from the National Institutes of Health-R21 AI121997(to HNS)and R21 AI144738-01A1(to CS)and by the Nutrition Obesity Research Center at Harvard(P30 DK040561)CS was supported by a Pilot Feasibility Grant from the Nutrition Obesity Research Center at Harvard(P30 DK040561)LJ and TM were sponsored by the China Scholarship Council.The funders had no role in the study design,collection,analysis,or interpretation of data.
文摘Helminth-induced Th2 immunity and gut microbiota have been recently shown to be highly effective in modulating metabolic syndromes in animal models.This study aimed to determine whether maternal immunity and microbial factors affect the induction and development of obesity in offspring.Here,Heligomosomoides polygyrus(Hp)-infected or control female C57BL/6J mice mated with normal males and their offspring were fed a high-fat diet(HFD)for 9 weeks after weaning.Our results showed that Hp-induced maternal outcomes during gestation and lactation significantly impacted offspring metabolic phenotypes.This was evidenced by results showing that offspring from helminth-infected mothers on an HFD(Hp-offspring+HFD)gained significantly less body weight than those from uninfected mothers(Cont-offspring+HFD).Hp-offspring+HFD exhibited no Th2 phenotype but displayed a pattern of gut microbiota composition similar to that of Hp-infected mothers.Cross-fostering experiments confirmed that the helminth-induced maternal attenuation of offspring obesity was mediated through both prenatal and postnatal effects.Our results further showed that helminth-infected dams and their offspring had a markedly altered gut microbiome composition,with increased production of short-chain fatty acids(SCFAs).Intriguingly,Hp-infected mothers and Hp-offspring+HFD showed increased SCFA receptor(GPR)expression in adipose and colonic tissues compared to noninfected mothers and Cont-offspring+HFD,respectively.Moreover,SCFA supplementation to the pups of uninfected control mothers during lactation protected against HFD-induced weight gain,which corresponded with changes in gut bacterial colonization.Collectively,our findings provide new insights into the complex interaction of maternal immune status and gut microbiome,Hp infection,and the immunity and gut microbiome in obese-prone offspring in infant life.
基金This work was supported by the National Natural Science Foundation of China(41876166)Natural Science Foundation of Shandong Province,China(Grant No.ZR2022QD126)Science and Technology Fundamental Resources Investigation Program(Grant No.2022FY101100,2019FY100700).
文摘Many marine bacteria are difcult to culture because they are dormant,rare or found in low-abundances.Enrichment culturing has been widely tested as an important strategy to isolate rare or dormant microbes.However,many more mechanisms remain uncertain.Here,based on 16S rRNA gene high-throughput sequencing and metabolomics technology,it was found that the short-chain fatty acids(SCFAs)in metabolites were signifcantly correlated with uncultured bacterial groups during enrichment cultures.A pure culture analysis showed that the addition of SCFAs to media also resulted in high efciency for the isolation of uncultured strains from marine sediments.As a result,238 strains belonging to 10 phyla,26 families and 82 species were successfully isolated.Some uncultured rare taxa within Chlorobi and Kiritimatiellaeota were successfully cultured.Amongst the newly isolated uncultured microbes,most genomes,e.g.bacteria,possess SCFA oxidative degradation genes,and these features might aid these microbes in better adapting to the culture media.A further resuscitation analysis of a viable but non-culturable(VBNC)Marinilabiliales strain verifed that the addition of SCFAs could break the dormancy of Marinilabiliales in 5 days,and the growth curve test showed that the SCFAs could shorten the lag phase and increase the growth rate.Overall,this study provides new insights into SCFAs,which were frst studied as resuscitation factors in uncultured marine bacteria.Thus,this study can help improve the utilisation and excavation of marine microbial resources,especially for the most-wanted or key players.
