丁酸梭菌和凝结芽孢杆菌对虹鳟生长性能、肝功能及肠道菌群的影响
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摘要
为研究饲料中添加丁酸梭菌Clostridium butyricum(CGMCC1.336)、凝结芽孢杆菌Bacillus coagulans(CGMCC1.3220)对虹鳟Oncorhynchus mykiss生长性能、肝功能及肠道菌群的影响,试验设T1、T2、T3 3个益生菌处理组,分别投喂基础饲料中添加丁酸梭菌(7.35×10~9 CFU/kg)、凝结芽孢杆菌(2.4×10~9 CFU/kg)和丁酸梭菌+凝结芽孢杆菌(7.35×10~9 CFU/kg+2.4×10~9 CFU/kg)的试验饲料,对照组(T0)投喂基础饲料,每组设3个重复,每个重复放20尾鱼,将初始体质量为(73.0±2.0)g的虹鳟饲养于水族箱(80 cm×50 cm×60 cm)中,养殖试验共进行40 d。结果表明:饲料中添加丁酸梭菌、凝结芽孢杆菌均能显著提高虹鳟的特定生长率(SGR)(P<0.05),T1和T3组SGR最高,均达1.41%/d,而饲料系数(FCR)则较低,分别为1.08和1.10;T1和T3组虹鳟体成分粗蛋白质含量显著高于对照组(P<0.05),各组粗灰分和水分含量无显著性差异(P>0.05);各益生菌组虹鳟肝脏谷草转氨酶(GOT)、γ-谷氨酰转肽酶(γ-GT)活性、总胆红素(TBiL)含量和肝指数均显著低于对照组(P<0.05),T3组效果最好,而胆碱酯酶(CHE)活性则显著高于对照组(P<0.05),其中T2、T3组效果较好;T3组肠道菌群的Chao1、Simpson、Shannon指数较对照组增加最显著(P<0.05);从肠道菌群结构的门水平上看,T2和T3组变形菌门比例明显降低,而厚壁菌门、拟杆菌门比例则明显提高,T1和T3组梭杆菌门比例明显提高。研究表明,饲料中添加丁酸梭菌、凝结芽孢杆菌可促进虹鳟生长,改善其机体营养成分,维护肝功能,改善肠道菌群结构。
Rainbow trout Oncorhynchus mykiss with average body weight of(73.0±2.0) g were reared in aquarium(80 cm×50 cm×60 cm) and fed basal diet as control group(T0) at stocking density of 20 individuals per aquarium, and the basal diet supplemented with Clostridium butyricum(7.35×10~9 CFU/kg, T1), Bacillus coagulans(2.4×10~9 CFU/kg, T2), and Clostridium butyricum+ Bacillus coagulans(7.35×10~9 CFU/kg+2.4×10~9 CFU/kg, T3) for 40 days in order to investigate the effects of Clostridium butyricum(CGMCC1.336) and Bacillus coagulans(CGMCC1.3220) on growth performance, hepatic function and intestinal microbiota of rainbow trout with 3 replicates. The specific growth rate(SGR) were found to be significantly higher in the fish fed the diets containing Clostridium butyricum and Bacillus coagulans(P<0.05), with the maximal SGR in groups T1 and T3(1.41%/d)(P<0.05), and with food conversion ratio(FCR) of 1.08 and 1.10, respectively(P<0.05). There was significantly higher crude protein in groups T1 and T3 than that in the control(P<0.05), without significant differences in the crude ash content and moisture in all groups(P>0.05). There were significantly lower activities of glutamic oxaloacetic transaminase(GOT), and γ-glutamyl transpeptidase(γ-GT), total bilirubin content(TBiL) and hepato-somatic index in live in T3 group(P<0.05), with significantly higher activity of cholinesterase(CHE) in groups T2 and T3(P<0.05). The indices of Chao1, Simpson and Shannon were increased in T3(P<0.05), meanwhile the phylum level of bacterial community was changed, with significant decrease in proportion of Proteobacteria in T2 and T3, and with increase in proportions of Firmicutes, Bacteroidetes and Fusobacteria in T1 and T3. The findings indicate that dietary C.butyricum and B.coagulans leads to promote the growth performance and body composition, to protect the hepatic function, and to optimize the intestinal microbiota in rainbow trout.
Rainbow trout Oncorhynchus mykiss with average body weight of(73.0±2.0) g were reared in aquarium(80 cm×50 cm×60 cm) and fed basal diet as control group(T0) at stocking density of 20 individuals per aquarium, and the basal diet supplemented with Clostridium butyricum(7.35×10~9 CFU/kg, T1), Bacillus coagulans(2.4×10~9 CFU/kg, T2), and Clostridium butyricum+ Bacillus coagulans(7.35×10~9 CFU/kg+2.4×10~9 CFU/kg, T3) for 40 days in order to investigate the effects of Clostridium butyricum(CGMCC1.336) and Bacillus coagulans(CGMCC1.3220) on growth performance, hepatic function and intestinal microbiota of rainbow trout with 3 replicates. The specific growth rate(SGR) were found to be significantly higher in the fish fed the diets containing Clostridium butyricum and Bacillus coagulans(P<0.05), with the maximal SGR in groups T1 and T3(1.41%/d)(P<0.05), and with food conversion ratio(FCR) of 1.08 and 1.10, respectively(P<0.05). There was significantly higher crude protein in groups T1 and T3 than that in the control(P<0.05), without significant differences in the crude ash content and moisture in all groups(P>0.05). There were significantly lower activities of glutamic oxaloacetic transaminase(GOT), and γ-glutamyl transpeptidase(γ-GT), total bilirubin content(TBiL) and hepato-somatic index in live in T3 group(P<0.05), with significantly higher activity of cholinesterase(CHE) in groups T2 and T3(P<0.05). The indices of Chao1, Simpson and Shannon were increased in T3(P<0.05), meanwhile the phylum level of bacterial community was changed, with significant decrease in proportion of Proteobacteria in T2 and T3, and with increase in proportions of Firmicutes, Bacteroidetes and Fusobacteria in T1 and T3. The findings indicate that dietary C.butyricum and B.coagulans leads to promote the growth performance and body composition, to protect the hepatic function, and to optimize the intestinal microbiota in rainbow trout.
引文
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[9] 何伟聪,董晓慧,谭北平,等.益生菌对军曹鱼幼鱼生长性能、消化酶和免疫酶活性的影响[J].动物营养学报,2015,27(12):3821-3830.
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[11] 华洵璐,钱仁界,张宪中,等.芽孢杆菌复合制剂对断奶仔猪生长、腹泻和饲料消化率的影响[J].安徽农业科学,2015,43(3):166-170.
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[18] 曲亚玲,袁莉刚,钱续,等.中草药复合添加剂对虹鳟鱼增重及肝功能和结构的影响[J].中国兽医科学,2012,42(8):865-869.
[19] 王珊珊,王佳堃,刘建新.肠道微生物对宿主免疫系统的调节及其可能机制[J].动物营养学报,2015,27(2):375-382.
[20] Wang Xianghong,Li Huirong,Zhang Xiaohua,et al.Microbial flora in the digestive tract of adult penaeid shrimp(Penaeus chinensis)[J].Journal of Ocean University of Qingdao,2000,30(3):493-498.
[21] Li Jiqiu,Tan Beiping,Mai Kangsen.Dietary probiotic Bacillus OJ and isomaltooligosaccharides influence the intestine microbial populations,immune responses and resistance to white spot syndrome virus in shrimp (Litopenaeus vannamei)[J].Aquaculture,2009,291(1-2):35-40.
[22] Zhou Zhigang,Ding Zhaokun,Huiyuan L V.Effects of dietary short-chain fructooligosaccharides on intestinal microflora,survival,and growth performance of juvenile white shrimp,Litopenaeus vannamei[J].Journal of the World Aquaculture Society,2007,38(2):296-301.
[23] Rendueles O,Ferrières L,Frétaud M,et al.A new zebrafish model of Oro-intestinal pathogen colonization reveals a key role for adhesion in protection by probiotic bacteria[J].PLoS Pathogens,2012,8(7):e1002815.
[24] Hung A T,Lin Shuyuan,Yang T Y,et al.Effects of Bacillus coagulans ATCC7050 on growth performance,intestinal morphology,and microflora composition in broiler chickens[J].Animal Production Science,2012,52(9):874-879.
[25] 肖英平,杨彩梅,代兵,等.基于高通量测序的丁酸梭菌对肉鸡盲肠菌群结构的影响[J].浙江农业学报,2017,29(3):373-379.
[2] 王瑶,刘赛.微生物饲料添加剂的功能与安全性探讨[J].农业与技术,2016,36(16):22.
[3] Liu C H,Chiu C H,Wang Shiwei,et al.Dietary administration of the probiotic,Bacillus subtilis E20,enhances the growth,innate immune responses,and disease resistance of the grouper,Epinephelus coioides[J].Fish & Shellfish Immunology,2012,33(4):699-706.
[4] 夏汉钦,杨红玲,孙云章.活性和灭活嗜冷杆菌SE6对斜带石斑鱼肠道菌群和肠黏膜免疫基因表达的影响[C]//第九届世界华人鱼虾营养学术研讨会论文集.厦门:中国水产学会,2013.
[5] Zhang Qin,Ma Hongming,Mai Kangsen,et al.Interaction of dietary Bacillus subtilis and fructooligosaccharide on the growth performance,non-specific immunity of sea cucumber,Apostichopus japonicus[J].Fish & Shellfish Immunology,2010,29(2):204-211.
[6] Munir M B,Hashim R,Chai Y H,et al.Dietary prebiotics and probiotics influence growth performance,nutrient digestibility and the expression of immune regulatory genes in snakehead (Channa striata) fingerlings[J].Aquaculture,2016,460:59-68.
[7] 严芳,胡炜,李成林,等.饲料中添加活性酵母制剂对刺参生长、免疫力和抗病力的影响[J].动物营养学报,2017,29(2):583-589.
[8] Huang Lu,Ran Chao,He Suxu,et al.Effects of dietary Saccharomyces cerevisiae culture or live cells with Bacillus amyloliquefaciens spores on growth performance,gut mucosal morphology,hsp70 gene expression,and disease resistance of juvenile common carp (Cyprinus carpio)[J].Aquaculture,2015,438:33-38.
[9] 何伟聪,董晓慧,谭北平,等.益生菌对军曹鱼幼鱼生长性能、消化酶和免疫酶活性的影响[J].动物营养学报,2015,27(12):3821-3830.
[10] 王纯,孙国祥,刘志培,等.解淀粉芽孢杆菌和胶红酵母复合菌对虹鳟生长性能及胃黏膜、肠黏膜菌群结构的影响[J].中国水产科学,2017,24(4):746-756.
[11] 华洵璐,钱仁界,张宪中,等.芽孢杆菌复合制剂对断奶仔猪生长、腹泻和饲料消化率的影响[J].安徽农业科学,2015,43(3):166-170.
[12] 王俊东,刘宗平.兽医临床诊断学[M].2版.北京:中国农业出版社,2010:217-228.
[13] Páez-Martiíinez N,Cruz S L,López-Rubalcava C.Comparative study of the effects of toluene,benzene,1,1,1-trichloroethane,diethyl ether,and flurothyl on anxiety and nociception in mice[J].Toxicology and Applied Pharmacology,2003,193(1):9-16.
[14] 孙金辉,范泽,张美静,等.饲料蛋白水平对鲤幼鱼肝功能和抗氧化能力的影响[J].南方水产科学,2017,13(3):113-119.
[15] Malbrouck C,Trausch G,Devos P,et al.Hepatic accumulation and effects of microcystin-LR on juvenile goldfish Carassius auratus L.[J].Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology,2003,135(1):39-48.
[16] Anderson F H,Zeng Lecheng,Rock N R,et al.An assessment of the clinical utility of serum ALT and AST in chronic hepatitis C[J].Hepatology Research,2000,18(1):63-71.
[17] 贾艳菊,杨虹,侯金良,等.饲料中添加乌贼肝粉和鱼浆蛋白对中华鳖生长性能和肝功能的影响[J].浙江大学学报:农业与生命科学版,2014,40(3):348-354.
[18] 曲亚玲,袁莉刚,钱续,等.中草药复合添加剂对虹鳟鱼增重及肝功能和结构的影响[J].中国兽医科学,2012,42(8):865-869.
[19] 王珊珊,王佳堃,刘建新.肠道微生物对宿主免疫系统的调节及其可能机制[J].动物营养学报,2015,27(2):375-382.
[20] Wang Xianghong,Li Huirong,Zhang Xiaohua,et al.Microbial flora in the digestive tract of adult penaeid shrimp(Penaeus chinensis)[J].Journal of Ocean University of Qingdao,2000,30(3):493-498.
[21] Li Jiqiu,Tan Beiping,Mai Kangsen.Dietary probiotic Bacillus OJ and isomaltooligosaccharides influence the intestine microbial populations,immune responses and resistance to white spot syndrome virus in shrimp (Litopenaeus vannamei)[J].Aquaculture,2009,291(1-2):35-40.
[22] Zhou Zhigang,Ding Zhaokun,Huiyuan L V.Effects of dietary short-chain fructooligosaccharides on intestinal microflora,survival,and growth performance of juvenile white shrimp,Litopenaeus vannamei[J].Journal of the World Aquaculture Society,2007,38(2):296-301.
[23] Rendueles O,Ferrières L,Frétaud M,et al.A new zebrafish model of Oro-intestinal pathogen colonization reveals a key role for adhesion in protection by probiotic bacteria[J].PLoS Pathogens,2012,8(7):e1002815.
[24] Hung A T,Lin Shuyuan,Yang T Y,et al.Effects of Bacillus coagulans ATCC7050 on growth performance,intestinal morphology,and microflora composition in broiler chickens[J].Animal Production Science,2012,52(9):874-879.
[25] 肖英平,杨彩梅,代兵,等.基于高通量测序的丁酸梭菌对肉鸡盲肠菌群结构的影响[J].浙江农业学报,2017,29(3):373-379.