This study was conducted to evaluate the effects of dietary crude protein(CP)and rumen-protected lysine(RPL)supplementation on lactation performance,amino acid(AA)balance,nitrogen(N)utilization and hindgut microbiota ...This study was conducted to evaluate the effects of dietary crude protein(CP)and rumen-protected lysine(RPL)supplementation on lactation performance,amino acid(AA)balance,nitrogen(N)utilization and hindgut microbiota in dairy cows.Treatments were in a 2×2 factorial arrangement,and the main effects were CP concentration(16%vs.18%)and RPL supplementation(with or without RPL at40 g/cow per day).Forty cows were randomly allocated to 4 groups:low-CP diet(LP),low-CP diet plus RPL(LPL),high-CP diet(HP),high-CP diet plus RPL(HPL).The experiment was conducted for 8 weeks.Results showed that RPL increased the dry matter intake(P<0.01),milk protein yield(P=0.04)and energy corrected milk(P=0.04),and tended to increase milk fat yield(P=0.06)and fat corrected milk(P=0.05).Cows in the HP group tended to have higher milk urea N(P=0.07).Plasma concentrations of Arg,Ile,Lys,Met,Pro,total essential AA and total nonessential AA were increased by RPL(P<0.05).The total essential AA,total nonessential AA and most AA(except Ile,Phe,Gly and Pro)were increased in the HP group(P<0.05).N excretion was increased in the HP group through an increase in urea N excretion(P<0.01)and an upward trend in plasma urea N(P=0.07).In addition,RPL tended to increase milk protein N secretion(P=0.08),milk N(P=0.07)and microbial protein synthesis(P=0.06),and decreased plasma urea N(P<0.001).In the hindgut,the bacterial community were different between the LP and LPL groups(P<0.01).The probiotic abundances of Christensenellaceae_R-7_group and Acinetobacter were increased by RPL(P=0.03 and 0.03,respectively).The pathogenic abundances of Clostridium_sensu_stricto_1(P<0.001)and Turicibacter(P<0.01)were decreased by RPL.In conclusion,supplementing RPL with low dietary CP could balance AA supply and increase milk protein yield,resulting in an improvement in N utilization efficiency,and altered the composition of the hindgut microbiota to favor the lactation performance of dairy cows.展开更多
In forested stream ecosystems of the north and eastern United States, larvae of the aquatic crane fly Tipula abdominalis are important shredders of leaf litter detritus. T. abdominalis larvae harbor a dense and divers...In forested stream ecosystems of the north and eastern United States, larvae of the aquatic crane fly Tipula abdominalis are important shredders of leaf litter detritus. T. abdominalis larvae harbor a dense and diverse microbial community in their hindgut that may aide in the degradation oflignocellulose. In this study, the activities ofcellulolytic and hemicellulolytic enzymes were demonstrated from hindgut extracts and from bacterial isolates using model sugar substrates. One of the bacterial isolates was further characterized as a member of the family Microbacteriaceae. Taxonomic position of the isolate within this family was determined by a polyphasic approach, as is commonly employed for the separation of genera within the family Microbacteriaceae. The bacterial isolate is Gramtype positive, motile, non-sporulating, and rod-shaped. The G + C content of the DNA is 64.9 mol%. The cell wall contains B2y type peptidoglycan, D- and L-diaminobutyric acid as the diamino acid, and rhamnose as the predominant sugar. The predominant fatty acids are 12-methyltetradecanoic acid (ai-C15:0) and 14-methylhexadecanoic acid (ai-C17:0). The isolate forms a distinct lineage within the family Microbacteriaceae, as determined by 16S rRNA sequence analysis. We propose the name Crocebacterium ilecola gen. nov., sp. nov., to accommodate this bacterial isolate. The type species is T202T (ATCC BAA-1359; GenBank Accession DQ826511).展开更多
Early weaning induces intestinal injury,leading to a series of long-term symptoms such as inflammation,malabsorption and diarrhea.In this study,we hypothesized that microbes and theirmetabolitesmodulate the host's...Early weaning induces intestinal injury,leading to a series of long-term symptoms such as inflammation,malabsorption and diarrhea.In this study,we hypothesized that microbes and theirmetabolitesmodulate the host's inflammatory response to early weaning stress in a goatmodel.A total of 18 female Tibetan goat kids(n?9)wereweaned fromtheirmothers at 28 d(D28)and 60 d(D60)postpartum.D60 and D28 groupswere fed the same solid diet ad libitum fromweaning to 75 d of age.The colonic epithelium was subject to RNAsequencing,the caecal digesta metabolomics were assessed by liquid chromatographyetandem mass spectrometry(LC-MS/MS),and the caecal microbiota composition was analysed by 16S ribosomal RNA gene sequencing.We foundthatearlyweaningsubstantially increased the colonic pro-apoptotic gene expressionof B-cell lymphoma associated X(Bax),caspase-9,and caspase-3,and decreased the expression of zonula occludens-1(ZO-1)and claudin-1(P<0.01).In addition,a significant Bacteroides acidifaciens enrichmentwas observed in the hindgut of early-weaned goats(P<0.01),which negatively correlated with lysophosphatidylcholine products.Similarly,the chemokine signaling,IL-17 signaling,and peroxisome proliferatorsactivated receptor(PPAR)signaling pathways were upregulated in the colonic mucosa of the early-weaned goats.By applying caecal microbiota transplantation from goats to defaunated C57/6J mice,we confirmed that caecalmicrobiota of D28 goat kids increased the relative abundance of B.acidifaciens and significantly upregulated the genes of Bax,G proteinecoupled receptor(GPR)109A,GPR 43,fatty acid binding protein 6,nuclear receptor subfamily 1 group H member 3,angiotensin converting enzyme 2,and IL-6 expression(P<0.05),and decreased ZO-1,and claudin-1 protein expression in the mice jejunum and colon(P<0.001).These results proposed that the hindgut microbiota andmetabolites mediate the barrier functionweakening duringearlyweaning,and the relative abundance of B.acidifacienswas negatively correlatedwiththe hindgut barrier gene expression.This studydemonstrateshowweaningstress canaffectkeyhostemicrobe interaction regulators in the hindgut,in a lysophosphatidylcholine dependent and independent manner.Furthermore,based on our mice data,these results are transferable to other mammal species.展开更多
Chitin deacetylases(CDAs,including CDAI and CDA2)are considered key enzymes for body cuticle formation and tracheal morphogenesis in various insect species.However,their functions in the formation of the cuticular int...Chitin deacetylases(CDAs,including CDAI and CDA2)are considered key enzymes for body cuticle formation and tracheal morphogenesis in various insect species.However,their functions in the formation of the cuticular intima of the foregut and hindgut are unclear.Here,we investigated the roles of their respective genes LmCDAI and Lm-CD42 in this process,in the hemimetabolous insect Locusta migratoria.Transcripts of LmCDAI and LmCDA2 were highly expressed both before and after molting in the foregut.In the hindgut,their expression was high only before molting.In both the foregut and hindgut,LmCDA1 protein was localized in the basal half of the chitin matrix(procuti-cle),whereas LmCDA2 was detected in the upper half of the procuticle.K nockdown of LmCDAI by RNA interference(RNAi)in 5th-instar nymphs caused no visible defcts of the hindgut cuticle.By contrast,the chitinous lamellae of the cuticular intima in the foregut of knockdown animals were less compact than in control animals.RNAi against LmCDA2 led to thickening of both the foregut and hindgut cuticles,with a greater number of thinner laminae than in the respective control cuticles.Taken together,our results show that LmCDAI and LmCDA2 have distinct,but overlapping,functions in chitin organization in the foregut cuticle.However,in the hindgut,this process seems independent of LmCDA1 activity but requires LmCDA2 function.Thus,the CDAs reflect tissue-specific differences in cuticular organization and function,which need further detailed molecular and histological analyses for full comprehension.展开更多
采用Bead beating法和QIAamp DNA stool mini kit法提取蝗虫肠道微生物总DNA,并对2种方法提取DNA的得率、完整性以及16SrRNA基因扩增产物的变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)图谱等进行综合比较。结果表...采用Bead beating法和QIAamp DNA stool mini kit法提取蝗虫肠道微生物总DNA,并对2种方法提取DNA的得率、完整性以及16SrRNA基因扩增产物的变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)图谱等进行综合比较。结果表明,Bead beating法提取DNA的得率显著高于QIAamp DNA stool mini kit法(P=0.042),而QIAamp DNA stool mini kit法提取DNA片段更完整。PCR-DGGE检测微生物多样性结果显示,QIAamp DNA stool mini kit法提取DNA所代表的微生物群落多样性略高于Bead beating法,但Mann-Whitley统计学检验表明用2种方法检测蝗虫肠道微生物多样性无显著差异(P=0.17)。因此在蝗虫肠道微生物群落多样性的检测中QIAamp DNA stool mini kit法具一定的优势,而Bead beating法同样适用。展开更多
在常温下(25℃)解剖华山松大小蠹(Dendroctonus armandi Tsai et Li)的雌、雄虫,以正己烷为溶剂提取华山松大小蠹的后肠和粪便挥发物,并进行GC-MS分析。结果表明:在室温状态下,雌虫后肠挥发物中含有23种物质,主要为萜酸类、萜烯类、雌...在常温下(25℃)解剖华山松大小蠹(Dendroctonus armandi Tsai et Li)的雌、雄虫,以正己烷为溶剂提取华山松大小蠹的后肠和粪便挥发物,并进行GC-MS分析。结果表明:在室温状态下,雌虫后肠挥发物中含有23种物质,主要为萜酸类、萜烯类、雌雄甾类、醇类和醛类等;雄虫后肠中有25种,以有机酸(树脂型萜酸居多)、酯类化合物和萜烯类化合物为其主要成分;粪便中有33种,树脂型的萜酸最多。华山松大小蠹的化学活性物质以萜类化合物为主。展开更多
基金financially supported by National Natural Science Foundation of China (32172742 and 32202688)the Natural Science Foundation of Zhejiang Province (LQ22C170002)+1 种基金the Zhejiang Provincial Science and Technology Plan Project(2022C04038)the Zhejiang Animal Husbandry Industry Technology Project
文摘This study was conducted to evaluate the effects of dietary crude protein(CP)and rumen-protected lysine(RPL)supplementation on lactation performance,amino acid(AA)balance,nitrogen(N)utilization and hindgut microbiota in dairy cows.Treatments were in a 2×2 factorial arrangement,and the main effects were CP concentration(16%vs.18%)and RPL supplementation(with or without RPL at40 g/cow per day).Forty cows were randomly allocated to 4 groups:low-CP diet(LP),low-CP diet plus RPL(LPL),high-CP diet(HP),high-CP diet plus RPL(HPL).The experiment was conducted for 8 weeks.Results showed that RPL increased the dry matter intake(P<0.01),milk protein yield(P=0.04)and energy corrected milk(P=0.04),and tended to increase milk fat yield(P=0.06)and fat corrected milk(P=0.05).Cows in the HP group tended to have higher milk urea N(P=0.07).Plasma concentrations of Arg,Ile,Lys,Met,Pro,total essential AA and total nonessential AA were increased by RPL(P<0.05).The total essential AA,total nonessential AA and most AA(except Ile,Phe,Gly and Pro)were increased in the HP group(P<0.05).N excretion was increased in the HP group through an increase in urea N excretion(P<0.01)and an upward trend in plasma urea N(P=0.07).In addition,RPL tended to increase milk protein N secretion(P=0.08),milk N(P=0.07)and microbial protein synthesis(P=0.06),and decreased plasma urea N(P<0.001).In the hindgut,the bacterial community were different between the LP and LPL groups(P<0.01).The probiotic abundances of Christensenellaceae_R-7_group and Acinetobacter were increased by RPL(P=0.03 and 0.03,respectively).The pathogenic abundances of Clostridium_sensu_stricto_1(P<0.001)and Turicibacter(P<0.01)were decreased by RPL.In conclusion,supplementing RPL with low dietary CP could balance AA supply and increase milk protein yield,resulting in an improvement in N utilization efficiency,and altered the composition of the hindgut microbiota to favor the lactation performance of dairy cows.
文摘In forested stream ecosystems of the north and eastern United States, larvae of the aquatic crane fly Tipula abdominalis are important shredders of leaf litter detritus. T. abdominalis larvae harbor a dense and diverse microbial community in their hindgut that may aide in the degradation oflignocellulose. In this study, the activities ofcellulolytic and hemicellulolytic enzymes were demonstrated from hindgut extracts and from bacterial isolates using model sugar substrates. One of the bacterial isolates was further characterized as a member of the family Microbacteriaceae. Taxonomic position of the isolate within this family was determined by a polyphasic approach, as is commonly employed for the separation of genera within the family Microbacteriaceae. The bacterial isolate is Gramtype positive, motile, non-sporulating, and rod-shaped. The G + C content of the DNA is 64.9 mol%. The cell wall contains B2y type peptidoglycan, D- and L-diaminobutyric acid as the diamino acid, and rhamnose as the predominant sugar. The predominant fatty acids are 12-methyltetradecanoic acid (ai-C15:0) and 14-methylhexadecanoic acid (ai-C17:0). The isolate forms a distinct lineage within the family Microbacteriaceae, as determined by 16S rRNA sequence analysis. We propose the name Crocebacterium ilecola gen. nov., sp. nov., to accommodate this bacterial isolate. The type species is T202T (ATCC BAA-1359; GenBank Accession DQ826511).
基金Qinghai Province Key R&D and Transformation Plan(2020-NK-127)Tibet Science and Technology Department's"13th Five-Year Plan"Major Agriculture Project(XZ201901NA02)+2 种基金the Key Technology R&D Program of Xinjiang Groups(2020AB016)Science and Technology Program of Tibet(XZ202001YD0017C)Agriculture Research System of China(CARS-39-12).
文摘Early weaning induces intestinal injury,leading to a series of long-term symptoms such as inflammation,malabsorption and diarrhea.In this study,we hypothesized that microbes and theirmetabolitesmodulate the host's inflammatory response to early weaning stress in a goatmodel.A total of 18 female Tibetan goat kids(n?9)wereweaned fromtheirmothers at 28 d(D28)and 60 d(D60)postpartum.D60 and D28 groupswere fed the same solid diet ad libitum fromweaning to 75 d of age.The colonic epithelium was subject to RNAsequencing,the caecal digesta metabolomics were assessed by liquid chromatographyetandem mass spectrometry(LC-MS/MS),and the caecal microbiota composition was analysed by 16S ribosomal RNA gene sequencing.We foundthatearlyweaningsubstantially increased the colonic pro-apoptotic gene expressionof B-cell lymphoma associated X(Bax),caspase-9,and caspase-3,and decreased the expression of zonula occludens-1(ZO-1)and claudin-1(P<0.01).In addition,a significant Bacteroides acidifaciens enrichmentwas observed in the hindgut of early-weaned goats(P<0.01),which negatively correlated with lysophosphatidylcholine products.Similarly,the chemokine signaling,IL-17 signaling,and peroxisome proliferatorsactivated receptor(PPAR)signaling pathways were upregulated in the colonic mucosa of the early-weaned goats.By applying caecal microbiota transplantation from goats to defaunated C57/6J mice,we confirmed that caecalmicrobiota of D28 goat kids increased the relative abundance of B.acidifaciens and significantly upregulated the genes of Bax,G proteinecoupled receptor(GPR)109A,GPR 43,fatty acid binding protein 6,nuclear receptor subfamily 1 group H member 3,angiotensin converting enzyme 2,and IL-6 expression(P<0.05),and decreased ZO-1,and claudin-1 protein expression in the mice jejunum and colon(P<0.001).These results proposed that the hindgut microbiota andmetabolites mediate the barrier functionweakening duringearlyweaning,and the relative abundance of B.acidifacienswas negatively correlatedwiththe hindgut barrier gene expression.This studydemonstrateshowweaningstress canaffectkeyhostemicrobe interaction regulators in the hindgut,in a lysophosphatidylcholine dependent and independent manner.Furthermore,based on our mice data,these results are transferable to other mammal species.
基金the National Natural Science Foundation of China(Grant No.31672364,31761133021,32072419,31701794,31801749)the Pro-gram for Top Young Academic Leaders of Higher Learn-ing Institutions of Shanxi(TYAL)2017+1 种基金Scientific and Technological Innovation Programs of Higher Edu-cation Institutions in Shanxi(2017104)BM's contribu-tion was supported by the German Research Foundation(DFG grant MO1714/10-1).
文摘Chitin deacetylases(CDAs,including CDAI and CDA2)are considered key enzymes for body cuticle formation and tracheal morphogenesis in various insect species.However,their functions in the formation of the cuticular intima of the foregut and hindgut are unclear.Here,we investigated the roles of their respective genes LmCDAI and Lm-CD42 in this process,in the hemimetabolous insect Locusta migratoria.Transcripts of LmCDAI and LmCDA2 were highly expressed both before and after molting in the foregut.In the hindgut,their expression was high only before molting.In both the foregut and hindgut,LmCDA1 protein was localized in the basal half of the chitin matrix(procuti-cle),whereas LmCDA2 was detected in the upper half of the procuticle.K nockdown of LmCDAI by RNA interference(RNAi)in 5th-instar nymphs caused no visible defcts of the hindgut cuticle.By contrast,the chitinous lamellae of the cuticular intima in the foregut of knockdown animals were less compact than in control animals.RNAi against LmCDA2 led to thickening of both the foregut and hindgut cuticles,with a greater number of thinner laminae than in the respective control cuticles.Taken together,our results show that LmCDAI and LmCDA2 have distinct,but overlapping,functions in chitin organization in the foregut cuticle.However,in the hindgut,this process seems independent of LmCDA1 activity but requires LmCDA2 function.Thus,the CDAs reflect tissue-specific differences in cuticular organization and function,which need further detailed molecular and histological analyses for full comprehension.
文摘采用Bead beating法和QIAamp DNA stool mini kit法提取蝗虫肠道微生物总DNA,并对2种方法提取DNA的得率、完整性以及16SrRNA基因扩增产物的变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)图谱等进行综合比较。结果表明,Bead beating法提取DNA的得率显著高于QIAamp DNA stool mini kit法(P=0.042),而QIAamp DNA stool mini kit法提取DNA片段更完整。PCR-DGGE检测微生物多样性结果显示,QIAamp DNA stool mini kit法提取DNA所代表的微生物群落多样性略高于Bead beating法,但Mann-Whitley统计学检验表明用2种方法检测蝗虫肠道微生物多样性无显著差异(P=0.17)。因此在蝗虫肠道微生物群落多样性的检测中QIAamp DNA stool mini kit法具一定的优势,而Bead beating法同样适用。
文摘在常温下(25℃)解剖华山松大小蠹(Dendroctonus armandi Tsai et Li)的雌、雄虫,以正己烷为溶剂提取华山松大小蠹的后肠和粪便挥发物,并进行GC-MS分析。结果表明:在室温状态下,雌虫后肠挥发物中含有23种物质,主要为萜酸类、萜烯类、雌雄甾类、醇类和醛类等;雄虫后肠中有25种,以有机酸(树脂型萜酸居多)、酯类化合物和萜烯类化合物为其主要成分;粪便中有33种,树脂型的萜酸最多。华山松大小蠹的化学活性物质以萜类化合物为主。