载锌凹凸棒石黏土对瘤胃体外发酵细菌多样性的影响（英文）
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摘要
【目的】研究载锌凹凸棒石黏土对瘤胃体外发酵细菌多样性的影响。【方法】本试验采用离子交换法制备载锌凹凸棒石黏土,利用16S rDNA测序技术分析了载锌凹凸棒石黏土对奶牛瘤胃体外发酵细菌菌群和多样性的影响。【结果】研究共获得1490959个有效序列和87662个OTUs。测序结果表明,与对照组相比,试验Ⅳ组中的细菌多样性在体外发酵24 h时提高,试验Ⅳ组中的细菌丰度在体外发酵48 h时提高。细菌菌群组成分析表明,60个样本中的优势菌门主要是厚壁菌门、拟杆菌门、变形菌门和黏胶球形菌门。与对照组相比,各试验组的厚壁菌门在体外发酵24 h和48 h时均显著增加(P<0.05),试验Ⅳ组中拟杆菌门在发酵48 h时显著降低(P<0.05);60个样本中共获得124个细菌菌属,其中对照组与试验组中普氏菌属含量均没有显著变化(P>0.05)。在体外发酵48 h时,试验Ⅳ组中密螺旋体属含量与对照组相比显著增加(P<0.05),而食物谷菌属和假丁酸弧菌属含量与对照组相比均显著降低(P<0.05)。【结论】载锌凹凸棒石黏土对奶牛瘤胃体外发酵中细菌多样性产生一定影响,其影响随发酵时间和添加剂量而不同。
[Objective] The aim of this experiment was to study the effects of zinc-bearing palygorskite(Zn-Pal) on rumen bacterial diversity in vitro. [Methods] We prepared Zn-Pal by the ion-exchange, and evaluated the compositions of bacterial communities in 60 samples based on 16 S rDNA genes. [Results] We obtained a total of 1490959 effective sequences and 87662 OTUs. The bacterial diversity in the treatment Ⅳ group increased at 24 h, and the abundance of the treatment Ⅳ group increased at 48 h. Bacterial community composition analysis shows that the dominant phyla were Bacteroidetes, Firmicutes, Proteobacteria and Lentisphaerae. Compared with the control group, Firmicutes in treatment groups significantly increased(P<0.05) at 24 h and 48 h, whereas Bacteroidetes in the treatment Ⅳ group was decreased(P<0.05) at 48 h. At the genus level, the sequences could be assigned to 124 different genera. The content of Prevotella had no significant difference between the control and treatments. The relative abundance of Treponema in the treatment Ⅳ group was significantly higher at 48 h than in the control group(P<0.05), whereas the relative abundance of Victivallis and Pseudobutyrivibrio in treatment Ⅳ was lower. [Conclusion] Zn-Pal may affect rumen fluid bacterial diversity in dairy cows, and the degrees of this influence varied with the dose and time of Zn-Pal.
[Objective] The aim of this experiment was to study the effects of zinc-bearing palygorskite(Zn-Pal) on rumen bacterial diversity in vitro. [Methods] We prepared Zn-Pal by the ion-exchange, and evaluated the compositions of bacterial communities in 60 samples based on 16 S rDNA genes. [Results] We obtained a total of 1490959 effective sequences and 87662 OTUs. The bacterial diversity in the treatment Ⅳ group increased at 24 h, and the abundance of the treatment Ⅳ group increased at 48 h. Bacterial community composition analysis shows that the dominant phyla were Bacteroidetes, Firmicutes, Proteobacteria and Lentisphaerae. Compared with the control group, Firmicutes in treatment groups significantly increased(P<0.05) at 24 h and 48 h, whereas Bacteroidetes in the treatment Ⅳ group was decreased(P<0.05) at 48 h. At the genus level, the sequences could be assigned to 124 different genera. The content of Prevotella had no significant difference between the control and treatments. The relative abundance of Treponema in the treatment Ⅳ group was significantly higher at 48 h than in the control group(P<0.05), whereas the relative abundance of Victivallis and Pseudobutyrivibrio in treatment Ⅳ was lower. [Conclusion] Zn-Pal may affect rumen fluid bacterial diversity in dairy cows, and the degrees of this influence varied with the dose and time of Zn-Pal.
引文
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[3]Rawls JF,Samuel BS,Gordon JI.Gnotobiotic zebrafish reveal evolutionarily conserved responses to the gut microbiota.Proceedings of the National Academy of Sciences of the United States of America,2004,101(13):4596-4601.
[4]Walker CF,Black RE.Zinc and the risk for infectious disease.Annual Review of Nutrition,2004,24:255-275.
[5]Kennedy DW,Bunting LD.Alterations in ruminal utilization of magnesium and zinc in lambs fed different ratios of concentrate:forage.International Journal for Vitamin and Nutrition Research,1991,61(1):67-71.
[6]Chen YP,Cheng YF,Li XH,Zhang H,Yang WL,Wen C,Zhou YM.Dietary palygorskite supplementation improves immunity,oxidative status,intestinal integrity,and barrier function of broilers at early age.Animal Feed Science and Technology,2016,219:200-209.
[7]Chalvatzi S,Kalamaki MS,Arsenos G,Fortomaris P.Dietary supplementation with the clay mineral palygorskite affects performance and beneficially modulates caecal microbiota in laying pullets.Journal of Applied Microbiology,2016,120(4):1033-1040.
[8]Zaid MRB,Hasan M,Khan AA.Attapulgite in the treatment of acute diarrhoea:a double-blind placebo-controlled study.Journal of Diarrhoeal Diseases Research,1995,13(1):44-46.
[9]Zhang JM,LüYF,Tang CH,Wang XQ.Effects of dietary supplementation with palygorskite on intestinal integrity in weaned piglets.Applied Clay Science,2013,86:185-189.
[10]Xu GL,Dong FQ,Zhu GP,Deng YQ,Feng QM,Li GW.Study on zinc-type antibacterial element material and its application in coating.Journal of Sichuan University(Engineering Science Edition),2005,37(1):52-56.(in Chinese)徐光亮,董发勤,朱桂平,邓跃全,冯启明,李国武.锌型抗菌基元材料及其在涂料中的应用研究.四川大学学报(工程科学版),2005,37(1):52-56.
[11]Luo YW,Wang LC,Guo F,Zhou YM,Wang T.Effects of Zinc-bearing attapulgite on growth performance,immunity and serum biochemical indexes of broilers.Journal of the Chinese Cereals and Oils Association,2008,23(3):131-135.(in Chinese)罗有文,王龙昌,郭芳,周岩民,王恬.载Zn2+凹凸棒石黏土对肉鸡生长、免疫和血液生化指标的影响.中国粮油学报,2008,23(3):131-135.
[12]Zhang ZF,Wang WB,Wang AQ.Highly effective removal of Methylene Blue using functionalized attapulgite via hydrothermal process.Journal of Environmental Sciences,2015,33(7):106-115.
[13]Hu CH,Gu LY,Luan ZS,Song J,Zhu K.Effects of montmorillonite-zinc oxide hybrid on performance,diarrhea,intestinal permeability and morphology of weanling pigs.Animal Feed Science and Technology,2012,177(1/2):108-115.
[14]Hu CH,Qian ZC,Song J,Luan ZS,Zuo AY.Effects of zinc oxide-montmorillonite hybrid on growth performance,intestinal structure,and function of broiler chicken.Poultry Science,2013,92(1):143-150.
[15]Tang ZG,Wen C,Li P,Wang T,Zhou YM.Effect of zinc-bearing zeolite clinoptilolite on growth performance,nutrient retention,digestive enzyme activities,and intestinal function of broiler chickens.Biological Trace Element Research,2014,158(1):51-57.
[16]Tang ZG,Wen C,Wang LC,Wang T,Zhou YM.Effects of zinc-bearing clinoptilolite on growth performance,cecal microflora and intestinal mucosal function of broiler chickens.Animal Feed Science and Technology,2014,189:98-106.
[17]Yan R,Zhang L,Yang X,Wen C,Zhou YM.Bioavailability evaluation of zinc-bearing palygorskite as a zinc source for broiler chickens.Applied Clay Science,2015,119:155-160.
[18]Qiao LH,Chen YP,Wen C,Zhou YM.Effects of natural and heat modified palygorskite supplementation on the laying performance,egg quality,intestinal morphology,digestive enzyme activity and pancreatic enzyme m RNA expression of laying hens.Applied Clay Science,2015,104:303-308.
[19]Theodorou MK,Williams BA,Dhanoa MS,Mcallan AB,France J.A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds.Animal Feed Science and Technology,1994,48(3/4):185-197.
[20]Hamady M,Walker JJ,Harris JK,Gold NJ,Knight R.Error-correcting barcoded primers for pyrosequencing hundreds of samples in multiplex.Nature Methods,2008,5(3):235-237.
[21]Edgar RC.UPARSE:highly accurate OTU sequences from microbial amplicon reads.Nature Methods,2013,10(10):996-998.
[22]Chao A.Nonparametric estimation of the number of classes in a population.Scandinavian Journal of Statistics,1984,11(4):265-270.
[23]Chao A,Lee SM.Estimating the number of classes via sample coverage.Journal of the American Statistical Association,1992,87(417):210-217.
[24]Brown KR.Diagnosis and management of anaerobic infections.Tropical Doctor,1977,7(3):111-114.
[25]Wang Q,Garrity GM,Tiedje JM,Cole JR.Na?ve Bayesian classifier for rapid assignment of r RNA sequences into the new bacterial taxonomy.Applied and Environmental Microbiology,2007,73(16):5261-5267.
[26]Quast C,Pruesse E,Yilmaz P,Gerken J,Schweer T,Yarza P,Peplies J,Gl?ckner FO.The SILVA ribosomal RNA gene database project:improved data processing and web-based tools.Nucleic Acids Research,2012,41(D1):D590-D596.
[27]Marchuk A,Rengasamy P.Clay behaviour in suspension is related to the ionicity of clay-cation bonds.Applied Clay Science,2011,53(4):754-759.
[28]Rusmin R,Sarkar B,Biswas B,Churchman J,Liu YJ,Naidu R.Structural,electrokinetic and surface properties of activated palygorskite for environmental application.Applied Clay Science,2016,134:95-102.
[29]Kittelmann S,Seedorf H,Walters WA,Clemente JC,Knight R,Gordon JI,Janssen PH.Simultaneous amplicon sequencing to explore co-occurrence patterns of bacterial,archaeal and eukaryotic microorganisms in rumen microbial communities.PLo S One,2013,8(2):e47879.
[30]Tajima K,Nonaka I,Higuchi K,Takusari N,Kurihara M,Takenaka A,Mitsumori M,Kajikawa H,Aminov RI.Influence of high temperature and humidity on rumen bacterial diversity in Holstein heifers.Anaerobe,2007,13(2):57-64.
[31]Evans NJ,Brown JM,Murray RD,Getty B,Birtles RJ,Hart CA,Carter SD.Characterization of novel bovine gastrointestinal tract Treponema isolates and comparison with bovine digital dermatitis treponemes.Applied and Environmental Microbiology,2011,77(1):138-147.
[32]Li XH,Zhu YP,Wen C,Zhou YM.Zinc desorption and antibacterial activity on E.coli K88 of Zinc-bearing Palygorskite.Non-Metallic Mines,2015,38(3):9-12.(in Chinese)李晓晗,朱玉萍,温超,周岩民.载锌凹凸棒石黏土中锌的解吸及抑菌作用研究.非金属矿,2015,38(3):9-12.
[33]Wang LC,Zhang TT,Wen C,Jiang ZY,Wang T,Zhou YM.Protective effects of zinc-bearing clinoptilolite on broilers challenged with Salmonella pullorum.Poultry Science,2012,91(8):1838-1845.
[34]Xia JL,Wang C,Liu XX.Research on antimicrobial agents and their mechanisms of actions.Journal of Central South University(Natural Science),2004,35(1):31-38.(in Chinese)夏金兰,王春,刘新星.抗菌剂及其抗菌机理.中南大学学报(自然科学版),2004,35(1):31-38.
[35]Wallace RJ,Onodera R,Cotta MA.Metabolism of nitrogen-containing compounds//Hobson PN,Stewart CS.The Rumen Microbial Ecosystem.Dordrecht,Netherlands:Springer,1997:283-328.
[36]Bekele AZ,Koike S,Kobayashi Y.Genetic diversity and diet specificity of ruminal Prevotella revealed by 16S r RNAgene-based analysis.FEMS Microbiology Letters,2010,305(1):49-57.
[37]Jami E,Mizrahi I.Similarity of the ruminal bacteria across individual lactating cows.Anaerobe,2012,18(3):338-343.
[38]Eryavuz A,Dehority BA.Effects of supplemental zinc concentration on cellulose digestion and cellulolytic and total bacterial numbers in vitro.Animal Feed Science and Technology,2009,151(3/4):175-183.
[39]Arelovich HM,Owens FN,Horn GW,Vizcarra JA.Effects of supplemental zinc and manganese on ruminal fermentation,forage intake,and digestion by cattle fed prairie hay and urea.Journal of Animal Science,2000,78(11):2972-2979.
[40]Vázquez-Armijo JF,Martínez-Tinajero JJ,López D,Salem AFZM,Rojo R.In vitro gas production and dry matter degradability of diets consumed by goats with or without copper and zinc supplementation.Biological Trace Element Research,2011,144(1/3):580-587.
[2]Mackie RI.Mutualistic fermentative digestion in the gastrointestinal tract:diversity and evolution.Integrative and Comparative Biology,2002,42(2):319-326.
[3]Rawls JF,Samuel BS,Gordon JI.Gnotobiotic zebrafish reveal evolutionarily conserved responses to the gut microbiota.Proceedings of the National Academy of Sciences of the United States of America,2004,101(13):4596-4601.
[4]Walker CF,Black RE.Zinc and the risk for infectious disease.Annual Review of Nutrition,2004,24:255-275.
[5]Kennedy DW,Bunting LD.Alterations in ruminal utilization of magnesium and zinc in lambs fed different ratios of concentrate:forage.International Journal for Vitamin and Nutrition Research,1991,61(1):67-71.
[6]Chen YP,Cheng YF,Li XH,Zhang H,Yang WL,Wen C,Zhou YM.Dietary palygorskite supplementation improves immunity,oxidative status,intestinal integrity,and barrier function of broilers at early age.Animal Feed Science and Technology,2016,219:200-209.
[7]Chalvatzi S,Kalamaki MS,Arsenos G,Fortomaris P.Dietary supplementation with the clay mineral palygorskite affects performance and beneficially modulates caecal microbiota in laying pullets.Journal of Applied Microbiology,2016,120(4):1033-1040.
[8]Zaid MRB,Hasan M,Khan AA.Attapulgite in the treatment of acute diarrhoea:a double-blind placebo-controlled study.Journal of Diarrhoeal Diseases Research,1995,13(1):44-46.
[9]Zhang JM,LüYF,Tang CH,Wang XQ.Effects of dietary supplementation with palygorskite on intestinal integrity in weaned piglets.Applied Clay Science,2013,86:185-189.
[10]Xu GL,Dong FQ,Zhu GP,Deng YQ,Feng QM,Li GW.Study on zinc-type antibacterial element material and its application in coating.Journal of Sichuan University(Engineering Science Edition),2005,37(1):52-56.(in Chinese)徐光亮,董发勤,朱桂平,邓跃全,冯启明,李国武.锌型抗菌基元材料及其在涂料中的应用研究.四川大学学报(工程科学版),2005,37(1):52-56.
[11]Luo YW,Wang LC,Guo F,Zhou YM,Wang T.Effects of Zinc-bearing attapulgite on growth performance,immunity and serum biochemical indexes of broilers.Journal of the Chinese Cereals and Oils Association,2008,23(3):131-135.(in Chinese)罗有文,王龙昌,郭芳,周岩民,王恬.载Zn2+凹凸棒石黏土对肉鸡生长、免疫和血液生化指标的影响.中国粮油学报,2008,23(3):131-135.
[12]Zhang ZF,Wang WB,Wang AQ.Highly effective removal of Methylene Blue using functionalized attapulgite via hydrothermal process.Journal of Environmental Sciences,2015,33(7):106-115.
[13]Hu CH,Gu LY,Luan ZS,Song J,Zhu K.Effects of montmorillonite-zinc oxide hybrid on performance,diarrhea,intestinal permeability and morphology of weanling pigs.Animal Feed Science and Technology,2012,177(1/2):108-115.
[14]Hu CH,Qian ZC,Song J,Luan ZS,Zuo AY.Effects of zinc oxide-montmorillonite hybrid on growth performance,intestinal structure,and function of broiler chicken.Poultry Science,2013,92(1):143-150.
[15]Tang ZG,Wen C,Li P,Wang T,Zhou YM.Effect of zinc-bearing zeolite clinoptilolite on growth performance,nutrient retention,digestive enzyme activities,and intestinal function of broiler chickens.Biological Trace Element Research,2014,158(1):51-57.
[16]Tang ZG,Wen C,Wang LC,Wang T,Zhou YM.Effects of zinc-bearing clinoptilolite on growth performance,cecal microflora and intestinal mucosal function of broiler chickens.Animal Feed Science and Technology,2014,189:98-106.
[17]Yan R,Zhang L,Yang X,Wen C,Zhou YM.Bioavailability evaluation of zinc-bearing palygorskite as a zinc source for broiler chickens.Applied Clay Science,2015,119:155-160.
[18]Qiao LH,Chen YP,Wen C,Zhou YM.Effects of natural and heat modified palygorskite supplementation on the laying performance,egg quality,intestinal morphology,digestive enzyme activity and pancreatic enzyme m RNA expression of laying hens.Applied Clay Science,2015,104:303-308.
[19]Theodorou MK,Williams BA,Dhanoa MS,Mcallan AB,France J.A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds.Animal Feed Science and Technology,1994,48(3/4):185-197.
[20]Hamady M,Walker JJ,Harris JK,Gold NJ,Knight R.Error-correcting barcoded primers for pyrosequencing hundreds of samples in multiplex.Nature Methods,2008,5(3):235-237.
[21]Edgar RC.UPARSE:highly accurate OTU sequences from microbial amplicon reads.Nature Methods,2013,10(10):996-998.
[22]Chao A.Nonparametric estimation of the number of classes in a population.Scandinavian Journal of Statistics,1984,11(4):265-270.
[23]Chao A,Lee SM.Estimating the number of classes via sample coverage.Journal of the American Statistical Association,1992,87(417):210-217.
[24]Brown KR.Diagnosis and management of anaerobic infections.Tropical Doctor,1977,7(3):111-114.
[25]Wang Q,Garrity GM,Tiedje JM,Cole JR.Na?ve Bayesian classifier for rapid assignment of r RNA sequences into the new bacterial taxonomy.Applied and Environmental Microbiology,2007,73(16):5261-5267.
[26]Quast C,Pruesse E,Yilmaz P,Gerken J,Schweer T,Yarza P,Peplies J,Gl?ckner FO.The SILVA ribosomal RNA gene database project:improved data processing and web-based tools.Nucleic Acids Research,2012,41(D1):D590-D596.
[27]Marchuk A,Rengasamy P.Clay behaviour in suspension is related to the ionicity of clay-cation bonds.Applied Clay Science,2011,53(4):754-759.
[28]Rusmin R,Sarkar B,Biswas B,Churchman J,Liu YJ,Naidu R.Structural,electrokinetic and surface properties of activated palygorskite for environmental application.Applied Clay Science,2016,134:95-102.
[29]Kittelmann S,Seedorf H,Walters WA,Clemente JC,Knight R,Gordon JI,Janssen PH.Simultaneous amplicon sequencing to explore co-occurrence patterns of bacterial,archaeal and eukaryotic microorganisms in rumen microbial communities.PLo S One,2013,8(2):e47879.
[30]Tajima K,Nonaka I,Higuchi K,Takusari N,Kurihara M,Takenaka A,Mitsumori M,Kajikawa H,Aminov RI.Influence of high temperature and humidity on rumen bacterial diversity in Holstein heifers.Anaerobe,2007,13(2):57-64.
[31]Evans NJ,Brown JM,Murray RD,Getty B,Birtles RJ,Hart CA,Carter SD.Characterization of novel bovine gastrointestinal tract Treponema isolates and comparison with bovine digital dermatitis treponemes.Applied and Environmental Microbiology,2011,77(1):138-147.
[32]Li XH,Zhu YP,Wen C,Zhou YM.Zinc desorption and antibacterial activity on E.coli K88 of Zinc-bearing Palygorskite.Non-Metallic Mines,2015,38(3):9-12.(in Chinese)李晓晗,朱玉萍,温超,周岩民.载锌凹凸棒石黏土中锌的解吸及抑菌作用研究.非金属矿,2015,38(3):9-12.
[33]Wang LC,Zhang TT,Wen C,Jiang ZY,Wang T,Zhou YM.Protective effects of zinc-bearing clinoptilolite on broilers challenged with Salmonella pullorum.Poultry Science,2012,91(8):1838-1845.
[34]Xia JL,Wang C,Liu XX.Research on antimicrobial agents and their mechanisms of actions.Journal of Central South University(Natural Science),2004,35(1):31-38.(in Chinese)夏金兰,王春,刘新星.抗菌剂及其抗菌机理.中南大学学报(自然科学版),2004,35(1):31-38.
[35]Wallace RJ,Onodera R,Cotta MA.Metabolism of nitrogen-containing compounds//Hobson PN,Stewart CS.The Rumen Microbial Ecosystem.Dordrecht,Netherlands:Springer,1997:283-328.
[36]Bekele AZ,Koike S,Kobayashi Y.Genetic diversity and diet specificity of ruminal Prevotella revealed by 16S r RNAgene-based analysis.FEMS Microbiology Letters,2010,305(1):49-57.
[37]Jami E,Mizrahi I.Similarity of the ruminal bacteria across individual lactating cows.Anaerobe,2012,18(3):338-343.
[38]Eryavuz A,Dehority BA.Effects of supplemental zinc concentration on cellulose digestion and cellulolytic and total bacterial numbers in vitro.Animal Feed Science and Technology,2009,151(3/4):175-183.
[39]Arelovich HM,Owens FN,Horn GW,Vizcarra JA.Effects of supplemental zinc and manganese on ruminal fermentation,forage intake,and digestion by cattle fed prairie hay and urea.Journal of Animal Science,2000,78(11):2972-2979.
[40]Vázquez-Armijo JF,Martínez-Tinajero JJ,López D,Salem AFZM,Rojo R.In vitro gas production and dry matter degradability of diets consumed by goats with or without copper and zinc supplementation.Biological Trace Element Research,2011,144(1/3):580-587.