脱氧雪腐镰刀菌烯醇与黄曲霉毒素B_1单一及联合染毒对小鼠肠道菌群的影响
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摘要
本试验旨在研究脱氧雪腐镰刀菌烯醇(DON)与黄曲霉毒素B1(AFB_1)单一及联合染毒对小鼠肠道菌群多样性、物种丰度的影响。选用18日龄昆明系雄性小鼠96只,随机分成4组,每组24只。其中,对照组灌服生理盐水,DON组灌服500μg/kg DON,AFB_1组灌服200μg/kg AFB_1,DON+AFB_1组灌服200μg/kg AFB_1+500μg/kg DON,连续灌服45 d。分别在试验第0天(试验开始前)、第45天时,无菌条件下采取直肠内新鲜粪便,每组随机采取3份,用Illumina HiSeq高通量测序技术分析肠道菌群结构和组成的变化。结果显示:与对照组相比,DON组、AFB_1组以及DON+AFB_1组拟杆菌门和拟杆菌纲的丰度均极显著低于对照组(P<0.01)。与DON组和AFB_1组相比,DON+AFB_1组拟杆菌纲的丰度极显著降低(P<0.01)。DON组、AFB_1组以及DON+AFB_1组脱铁杆菌门的丰度与对照组相比均极显著降低(P<0.01)。DON组、AFB_1组以及DON+AFB_1组梭菌纲的丰度极显著高于对照组(P<0.01)。DON组和DON+AFB_1组螺旋体门的丰度与对照组相比极显著升高(P<0.01)。综上所述,DON与AFB_1单一及联合染毒均可改变肠道内菌群的丰度,降低有益菌丰度,升高有害菌丰度,并且DON和AFB_1表现出明显的互作性,具有协同效应。
The present experiment was conducted to evaluate the effects of single and combined exposure of deoxynivalenol(DON) and aflatoxin B1(AFB_1) on intestinal microflora diversity and species abundance in mice.Ninety-six Kunming mice(male,18-day-old) were randomly divided into 4 groups with 24 mice in each group.Mice in control group were fed physiological saline by gavage.Mice in DON group were fed 500 μg/kg DON by gavage.Mice in AFB_1 group were fed 200 μg/kg AFB_1 by gavage.Mice in DON+AFB_1 group were fed 500 μg/kg DON+200 μg/kg AFB_1 by gavage.The trial was conducted for 45 d.On the days 0(before the experiment) and 45 of the experiment,the fresh dejections in the rectum from each group(each group was randomly taken 3 samples) were randomly taken under aseptic conditions,and then the changes of the structure and composition of the microbial community were analyzed using Illumina HiSeq high-throughput sequencing technology.The results showed that compared with the control group,the abundances of Bacteroidetes and Bacteroidia in DON,AFB_1 and DON+AFB_1 groups was significantly reduced(P<0.01),and the abundance of Bacteroidia in DON+AFB_1 group was significantly lower than that in DON and AFB_1 groups(P<0.01).Compared with the control group,the abundance of Deferribacteres in DON,AFB_1 and DON +AFB_1 groups was significantly reduced(P<0.01),while the abundance of Clostridia in DON,AFB_1 and DON + AFB_1 groups was significantly increased(P<0.01).Moreover,the abundance of Spirochaetes in DON and DON+AFB_1 groups was significantly higher than that in control group(P<0.01).In conclusion,single and combined exposure of DON and AFB_1 all can alter the abundances of intestinal microflora,decrease the abundances of beneficial bacteria,and increase the abundances of harmful bacteria.DON and AFB_1 exhibit a significant interaction with synergistic effects.
The present experiment was conducted to evaluate the effects of single and combined exposure of deoxynivalenol(DON) and aflatoxin B1(AFB_1) on intestinal microflora diversity and species abundance in mice.Ninety-six Kunming mice(male,18-day-old) were randomly divided into 4 groups with 24 mice in each group.Mice in control group were fed physiological saline by gavage.Mice in DON group were fed 500 μg/kg DON by gavage.Mice in AFB_1 group were fed 200 μg/kg AFB_1 by gavage.Mice in DON+AFB_1 group were fed 500 μg/kg DON+200 μg/kg AFB_1 by gavage.The trial was conducted for 45 d.On the days 0(before the experiment) and 45 of the experiment,the fresh dejections in the rectum from each group(each group was randomly taken 3 samples) were randomly taken under aseptic conditions,and then the changes of the structure and composition of the microbial community were analyzed using Illumina HiSeq high-throughput sequencing technology.The results showed that compared with the control group,the abundances of Bacteroidetes and Bacteroidia in DON,AFB_1 and DON+AFB_1 groups was significantly reduced(P<0.01),and the abundance of Bacteroidia in DON+AFB_1 group was significantly lower than that in DON and AFB_1 groups(P<0.01).Compared with the control group,the abundance of Deferribacteres in DON,AFB_1 and DON +AFB_1 groups was significantly reduced(P<0.01),while the abundance of Clostridia in DON,AFB_1 and DON + AFB_1 groups was significantly increased(P<0.01).Moreover,the abundance of Spirochaetes in DON and DON+AFB_1 groups was significantly higher than that in control group(P<0.01).In conclusion,single and combined exposure of DON and AFB_1 all can alter the abundances of intestinal microflora,decrease the abundances of beneficial bacteria,and increase the abundances of harmful bacteria.DON and AFB_1 exhibit a significant interaction with synergistic effects.
引文
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[2]XIAO H,WU M M,TAN B E,et al.Effects of composite antimicrobial peptides in weanling piglets challenged with deoxynivalenol:Ⅰ.Growth performance,immune function,and antioxidation capacity[J].Journal of Animal Science,2013,91(10):4772-4780.
[3]MANKEVICIENE6)A,JABLONSKYTE6)-RACE6)D,MAIKTE6)NIENE6)S.Occurrence of mycotoxins in spelt and common wheat grain and their products[J].Food Additives&Contaminants:Part A,2014,31(1):132-138.
[4]DUAN J L,YIN J,WU M M,et al.Dietary glutamate supplementation ameliorates mycotoxin-induced abnormalities in the intestinal structure and expression of amino acid transporters in young pigs[J].PLoS One,2013,9(11):e112357.
[5]LI X Y,ZHAO L H,FAN Y,et al.Occurrence of mycotoxins in feed ingredients and complete feeds obtained from the Beijing region of China[J].Journal of Animal Science and Biotechnology,2014,5(4):37.
[6]CAO X Q,WU S C,YUE Y,et al.A high-throughput method for the simultaneous determination of multiple mycotoxins in human and laboratory animal biological fluids and tissues by PLE and HPLC-MS/MS[J].Journal of Chromatography B,2013,942-943:113-125.
[7]李荣佳,李治忠,周闯,等.新型复合吸附剂HG对黄曲霉毒素B1和呕吐毒素的吸附脱毒研究[J].南京农业大学学报,2015,38(1):113-119.
[8]RIEDL R A,ATKINSON S N,BURNETT C M L,et al.The gut microbiome,energy homeostasis,and implications for hypertension[J].Current Hypertension Reports,2017,19(4):217.
[9]BELKAID Y,HAND T W.Role of the microbiota in immunity and inflammation[J].Cell,2014,157(1):121-141.
[10]ZHANG W,GU Y R,CHEN Y M,et al.Intestinal flora imbalance results in altered bacterial translocation and liver function in rats with experimental cirrhosis[J].European Journal of Gastroenterology&Hepatology,2010,22(12):1481-1486.
[11]LEE S Y,KIM H,KIM K,et al.Arhgap17,a RhoGTPase activating protein,regulates mucosal and epithelial barrier function in the mouse colon[J].Scientific Reports,2016,6:26923.
[12]程连平,舒迎霜,贺濛初,等.富硒酵母和枯草芽孢杆菌对湖羊羔羊小肠黏膜形态和直肠菌群的影响[J].动物营养学报,2018,30(11):4660-4669.
[13]FADROSH D W,MA B,GAJER P,et al.An improved dual-indexing approach for multiplexed 16SrRNA gene sequencing on the Illumina MiSeq platform[J].Microbiome,2014,2:6.
[14]MAGOCT,SALZBERG S L.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,2011,27(21):2957-2963.
[15]EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[16]TILG H,ADOLPH T E.Influence of the human intestinal microbiome on obesity and metabolic dysfunction[J].Current Opinion in Pediatrics,2015,27(4):496-501.
[17]POSSEMIERS S,BOLCA S,VERSTRAETE W,et al.The intestinal microbiome:a separate organ inside the body with the metabolic potential to influence the bioactivity of botanicals[J].Fitoterapia,2011,82(1):53-66.
[18]GERRITSEN J,SMIDT H,RIJKERS G T,et al.Intestinal microbiota in human health and disease:the impact of probiotics[J].Genes&Nutrition,2011,6:229.
[19]BLASER M J,KIRSCHNER D.The equilibria that allow bacterial persistence in human hosts[J].Nature,2007,449(7164):843-849.
[20]PETCHEY O L,EKLF A,BORRVALL C,et al.Trophically unique species are vulnerable to cascading extinction[J].The American Naturalist,2008,171(5):568-579.
[21]GILL S R,POP M,DEBOY R T,et al.Metagenomic analysis of the human distal gut microbiome[J].Science,2006,312(5778):1355-1359.
[22]WAIDMANN M,BECHTOLD O,FRICK J S,et al.Bacteroides vulgatus protects against Escherichia coliinduced colitis in gnotobiotic interleukin-2-deficient mice[J].Gastroenterology,2003,125(1):162-177.
[23]HOOPER L V,WONG M H,THELIN A,et al.Molecular analysis of commensal host-microbial relationships in the intestine[J].Science,2001,291(5505):881-884.
[24]BCKHED F,LEY R E,SONNENBURG J L,et al.Host-bacterial mutualism in the human intestine[J].Science,2005,307(5717):1915-1920.
[25]BCKHED F,DING H,WANG T,et al.The gut microbiota as an environmental factor that regulates fat storage[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(44):15718-15723.
[26]杨俊花,赵志辉,郭文博,等.应用Illumina-MiSeq高通量测序技术分析脱氧雪腐镰刀菌烯醇对小鼠肠道菌群的影响[J].动物营养学报,2017,29(1):158-167.
[27]WANG J C,TANG L L,GLENN T C,et al.Aflatoxin B1 induced compositional changes in gut microbial communities of male F344 rats[J].Toxicological Sciences,2015,150(1):54-63.
[28]SCALDAFERRI F,GERARDI V,LOPETUSO L R,et al.Gut gicrobial flora,prebiotics,and probiotics in IBD:their current usage and utility[J].Biomed Research International,2013,2013:435268.
[29]XIN J G,ZENG D,WANG H S,et al.Preventing nonalcoholic fatty liver disease through Lactobacillus johnsonii BS15 by attenuating inflammation and mitochondrial injury and improving gut environment in obese mice[J].Applied Microbiology and Biotechnology,2014,98(15):6517-6529.
[30]RITZE Y,BRDOS G,CLAUS A,et al.Lactobacillus rhamnosus GG protects against non-alcoholic fatty liver disease in mice[J].PLoS One,2014,9(1):e80169.
[31]KOCIOLEK L K,GERDING D N.Breakthroughs in the treatment and prevention of Clostridium difficile infection[J].Nature Reviews Gastroenterology&Hepatology,2016,13(3):150-160.
[32]MARIAT D,FIRMESSE O,LEVENEZ F,et al.The Firmicutes/Bacteroidetes ratio of the human microbiota changes with age[J].BMC Microbiology,2009,9:123.