卷曲乳杆菌对生长猪生长性能、粪便菌群和短链脂肪酸组成以及血清长链脂肪酸组成的影响
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摘要
本试验旨在研究猪源卷曲乳杆菌对生长猪生长性能、粪便菌群和短链脂肪酸组成以及血清长链脂肪酸组成的影响。选取体重(20.35±0.53) kg的二元杂交(长×大)生长猪120头,随机分成2组,每组4个重复,每个重复15头。对照组饲喂基础饲粮,试验组在基础饲粮中添加卷曲乳杆菌冻干制剂(卷曲乳杆菌添加量为5.5×109CFU/kg),饲喂期为30 d。饲养试验结束当天采集新鲜粪便用于16S rRNA V3～V4区菌群和短链脂肪酸组成的测定,采集血清用于长链脂肪酸组成的测定。结果显示:1)与对照组相比,试验组生长猪平均日增重提高了7. 32%(P <0.05),料重比降低了5.68%(P<0. 05); 2)饲粮中添加卷曲乳杆菌能够提高生长猪粪便菌群物种丰富度,增加厚壁菌门及乳酸杆菌属和光岗菌属的比例; 3)与对照组相比,试验组生长猪粪便中丁酸含量增加了6.20%(P<0.05),乙酸含量减少了9.88%(P<0.05); 4)试验组猪群血清中亚油酸和花生四烯酸含量分别比对照组提高了11.12%(P<0.05)和6.28%(P<0.05)。上述结果表明,在饲粮中添加5.5×109CFU/kg卷曲乳杆菌能够影响生长猪的粪便菌群组成,增加粪便中丁酸的含量,并改善血清中不饱和脂肪酸的组成,从而对生长猪的肠道健康及生长起到促进作用。
This experiment was conducted to study the effects of Lactobacillus crispatus on growth performance,fecal microbiota and short chain fatty acid composition and serum long chain fatty acid composition of growing pigs. A total of 120 two-way cross( Landrace×Yorkshire) growing pigs with an average body weight of( 20.35±0.53) kg were randomly distributed into 2 groups with 4 replicates per group and 15 pigs per replicate. Piglets in control group were fed a basal diet,and those in trial group were fed the basal diet supplemented with Lactobacillus crispatus preparation( the Lactobacillus crispatus supplemental level was 5. 5 × 109 CFU/kg). The experiment period was 30 d. Feces samples were collected and used for sequencing of the V3 to V4 hypervariable region of the 16S rRNA gene. Feces samples were also used to measure the short chain fatty acid composition and serum samples were analyzed for long chain fatty acid composition. The results showed as follows: 1) compared with the control group,the average daily gain( ADG) was increased by 7.32%( P < 0.05),and feed/gain( F/G) was decreased by 5.68%( P<0.05) in the trial group. 2) Dietary supplemented with Lactobacillus crispatus increased the microbiota richness,the Firmicutes at phylum,the Lactobacillus and Megasphaera at genus were also increased compared with the control group. 3) The content of butyric acid in feces was increased by 6.20%( P<0.05),and the content of acetic acid in feces was decreased by 9.88%( P<0.05) in the trial group compared with the control group. 4) The contents of serum C18 ∶2 n6 c and C20∶4 n6 were increased by 11.12%( P<0.05) and6.28%( P<0.05) in trial group,respectively. The results indicate that Lactobacillus crispatus has a beneficial effect on the intestinal microbiota composition,and increases the content of butyric acid in feces,improves serum long chain fatty acid composition,and then promotes the intestinal health and growth of growing pigs.
This experiment was conducted to study the effects of Lactobacillus crispatus on growth performance,fecal microbiota and short chain fatty acid composition and serum long chain fatty acid composition of growing pigs. A total of 120 two-way cross( Landrace×Yorkshire) growing pigs with an average body weight of( 20.35±0.53) kg were randomly distributed into 2 groups with 4 replicates per group and 15 pigs per replicate. Piglets in control group were fed a basal diet,and those in trial group were fed the basal diet supplemented with Lactobacillus crispatus preparation( the Lactobacillus crispatus supplemental level was 5. 5 × 109 CFU/kg). The experiment period was 30 d. Feces samples were collected and used for sequencing of the V3 to V4 hypervariable region of the 16S rRNA gene. Feces samples were also used to measure the short chain fatty acid composition and serum samples were analyzed for long chain fatty acid composition. The results showed as follows: 1) compared with the control group,the average daily gain( ADG) was increased by 7.32%( P < 0.05),and feed/gain( F/G) was decreased by 5.68%( P<0.05) in the trial group. 2) Dietary supplemented with Lactobacillus crispatus increased the microbiota richness,the Firmicutes at phylum,the Lactobacillus and Megasphaera at genus were also increased compared with the control group. 3) The content of butyric acid in feces was increased by 6.20%( P<0.05),and the content of acetic acid in feces was decreased by 9.88%( P<0.05) in the trial group compared with the control group. 4) The contents of serum C18 ∶2 n6 c and C20∶4 n6 were increased by 11.12%( P<0.05) and6.28%( P<0.05) in trial group,respectively. The results indicate that Lactobacillus crispatus has a beneficial effect on the intestinal microbiota composition,and increases the content of butyric acid in feces,improves serum long chain fatty acid composition,and then promotes the intestinal health and growth of growing pigs.
引文
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[14]LYBERG K,LUNDH T,PEDERSEN C.Influence of soaking,fermentation and phytase supplementation on nutrient digestibility in pigs offered a grower diet based on wheat and barley[J].Journal of Animal Science,2006,82(6):853-858.
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[16]LU X M,LU P Z,ZHANG H.Bacterial communities in manures of piglets and adult pigs bred with different feeds revealed by 16S r DNA 454 pyrosequencing[J].Applied Microbiology and Biotechnology,2014,98(6):2657-2665.
[17]王四新,季海峰,石国华,等.干酪乳杆菌对北京黑猪保育阶段生长性能及肠道菌群的影响[J].动物营养学报,2018,30(1):326-335.
[18]HILDEBRAND F,NGUYEN T,BRINKMAN B,et al.Inflammation-associated enterotypes,host genotype,cage and inter-individual effects drive gut microbiota variation in common laboratory mice[J].Genome Biology,2013,14:R4
[19]ZHAO W J,WANF Y P,LIU S Y,et al.The dynamic distribution of porcine microbiota across different ages and gastrointestinal tract segments[J].PLoS One,2015,10(2):e0117441,doi:10.1371/journal.pone.0117441.
[20]ARUMUGAM M,RAES J,PELLETIER E,et al.Enterotypes of the human gut microbiome[J].Nature,2011,473(7346):174-180.doi:10.1038/nature09944.
[21]LEY R E,TURNBAUGH P J,KLEIN S,et al.Microbial ecology:human gut microbes associated with obesity[J].Nature,2006,444(7122):1022-1023.
[22]PAJARILLO E A,CHAE J P,BALOLONG M P,et al.Pyrosequencing-based analysis of fecal microbial communities in three purebred pig lines[J].Journal of Microbiology,2014,52(8):646-651.
[23]ISAACSON R,KIM H B.The intestinal microbiome of the pig[J].Animal Health Research Reviews,2012,13(1):100-109.
[24]KASUBUCHI M,HASEGAWA S,HIRAMATSU T,et al.Dietary gut microbial metabolites,short-chain fatty acids,and host metabolic regulation[J].Nutrients,2015,7(4):2839-2849.
[25]KOH A,DE VADDER F,KOVATCHEVA-DATCHARYP,et al.From dietary fiber to host physiology:shortchain fatty acids as key bacterial metabolites[J].Cell,2016,165(6):1332-1345.
[26]DUNCAN S H,HOLTROP G,LOBLEY G E,et al.Contribution of acetate to butyrate formation by human faecal bacteria[J].British Journal of Nutrition,2004,91(6):915-923.
[27]ZHANG Y J,LIU Q,ZHANG W M,et al.Gastrointestinal microbial diversity and short-chain fatty acid production in pigs fed different fibrous diets with or without cell wall-degrading enzyme supplementation[J].Livestock Science,2017,207:105-116.
[28]MEEHAN C J,BEIKO R G.A phylogenomic view of ecological specialization in the Lachnospiraceae,a family of digestive tract associated bacteria[J].Genome Biology and Evolution,2014,6(3):703-713.
[29]LOUIS P,HOLD G L,FLINT H J.The gut microbiota,bacterial metabolites and colorectal cancer[J].Nature Reviews Microbiology,2014,12(10):661-672.
[30]FLINT H J,BAYER E A,RINCON M T,et al.Polysaccharide utilization by gut bacteria:potential for new insights from genomic analysis[J].Nature Reviews Microbiology,2008,6(2):121-131.
[31]PARK J Y,LEE S H,KIM K R,et al.Production of13S-hydroxy-9(Z)-octadecenoic acid from linoleic acid by whole recombinant cells expressing linoleate 13-hydratase from Lactobacillus acidophilus[J].Journal of Biotechnology,2015,208:1-10.
[32]ORTEGA-ANAY J,HERNNDEZ-SANTOYO A.Production of bioactive conjugated linoleic acid by the multifunctional enolase from Lactobacillus plantarum[J].International Journal of Biological Macromolecules,2016,91:524-535.
[33]ZHANG D Y,SHANG T T,HUANG Y,et al.Gene expression profile changes in the jejunum of weaned piglets after oral administration of Lactobacillus or an antibiotic[J].Scientific Reports,2017,7:15816,doi:10.1038/s41598-017-16158-y.
[34]S'WIATKIEWICZ M,OCZKOWICZ M,ROPKA-MOLIKK,et al.The effect of dietary fatty acid composition on adipose tissue quality and expression of genes related to lipid metabolism in porcine livers[J].Animal Feed Science and Technology,2016,216:204-215.
[2]LIU X T,HOU C L,ZHANG J,et al.Fermentation conditions influence the fatty acid composition of the membranes of Lactobacillus reuteri I5007 and its survival following freeze-drying[J].Letters in Applied Microbiology,2014,59(4):398-403.
[3]DEEPIKA G,CHARALAMPOPOULOS D.Surface and adhesion properties of Lactobacilli[J].Advances in Applied Microbiology,2010,70:127-152.
[4]DOWARAH R,VERMA A K,AGARWAL N,et al.Effect of swine based probiotic on performance,diarrhoea scores,intestinal microbiota and gut health of grower-finisher crossbred pigs[J].Livestock Science,2017,195:74-79.
[5]CHIANG M L,CHEN H C,CHEN K N,et al.Optimizing production of two potential probiotic Lactobacilli strains isolated from piglet feces as feed Additives for weaned piglets[J].Asian-Australasian Journal of Animal Science,2015,28(8):1163-1170.
[6]ZHANG D Y,JI H F,LIU H,et al.Changes in the diversity and composition of gut microbiota of weaned piglets after oral administration of Lactobacillus or an antibiotic[J].Applied Microbiology and Biotechnology,2016,100(23):10081-10093.
[7]SUN Z L,HUANG L H,KONG J,et al.In vitro evaluation of Lactobacillus crispatus K313 and K243:high-adhesion activity and anti-inflammatory effect on Salmonella braenderup infected intestinal epithelial cell[J].Veterinary Microbiology,2012,159(1/2):212-220.
[8]ABRAMOV V,KHLEBNIKOV V,KOSAREV I,et al.Probiotic properties of Lactobacillus crispatus 2,029:homeostatic interaction with cervicovaginal epithelial cells and antagonistic activity to genitourinary pathogens[J].Probiotics and Antimicrobial Proteins,2014,6(3/4):165-176.
[9]QIAO J Y,LI H H,WANG Z X,et al.Effects of Lactobacillus acidophilus dietary supplementation on the performance,intestinal barrier function,rectal microflora and serum immune function in weaned piglets challenged with Escherichia coli lipopolysaccharide[J].Antonie van Leeuwenhoek,2015,107(4):883-891.
[10]YEUNG C Y,CHIANG C J S,CHAN W T,et al.In vitro prevention of Salmonella lipopolysaccharide-induced damages in epithelial barrier function by various Lactobacillus strains[J].Gastroenterology Research and Practice,2013,2013:973209,doi:10.1155/2013/973209.
[11]肖驰,罗建模,周淑兰,等.日粮中添加乳酸菌制剂对生长猪的效果试验[J].养猪,1997(4):2-3.
[12]侯改凤.德氏乳杆菌对育肥猪生长性能、猪肉品质及脂肪沉积影响研究[D].硕士学位论文.长沙:湖南农业大学,2015.
[13]CANIBE N,JENSEN B B.Fermented and nonfermented liquid feed to growing pigs:effects on aspects of gastrointestinal ecology and growth performance[J].Journal of Animal Science,2003,81(8):2019-2031.
[14]LYBERG K,LUNDH T,PEDERSEN C.Influence of soaking,fermentation and phytase supplementation on nutrient digestibility in pigs offered a grower diet based on wheat and barley[J].Journal of Animal Science,2006,82(6):853-858.
[15]COLLINGTON G K,PARKER D S,ARMSTRONG DG.The influence of inclusion of either an antibiotic or a probiotic in the diet on the development of digestive enzyme activity in the pig[J].British Journal of Nutrition,1990,64(1):59-70.
[16]LU X M,LU P Z,ZHANG H.Bacterial communities in manures of piglets and adult pigs bred with different feeds revealed by 16S r DNA 454 pyrosequencing[J].Applied Microbiology and Biotechnology,2014,98(6):2657-2665.
[17]王四新,季海峰,石国华,等.干酪乳杆菌对北京黑猪保育阶段生长性能及肠道菌群的影响[J].动物营养学报,2018,30(1):326-335.
[18]HILDEBRAND F,NGUYEN T,BRINKMAN B,et al.Inflammation-associated enterotypes,host genotype,cage and inter-individual effects drive gut microbiota variation in common laboratory mice[J].Genome Biology,2013,14:R4
[19]ZHAO W J,WANF Y P,LIU S Y,et al.The dynamic distribution of porcine microbiota across different ages and gastrointestinal tract segments[J].PLoS One,2015,10(2):e0117441,doi:10.1371/journal.pone.0117441.
[20]ARUMUGAM M,RAES J,PELLETIER E,et al.Enterotypes of the human gut microbiome[J].Nature,2011,473(7346):174-180.doi:10.1038/nature09944.
[21]LEY R E,TURNBAUGH P J,KLEIN S,et al.Microbial ecology:human gut microbes associated with obesity[J].Nature,2006,444(7122):1022-1023.
[22]PAJARILLO E A,CHAE J P,BALOLONG M P,et al.Pyrosequencing-based analysis of fecal microbial communities in three purebred pig lines[J].Journal of Microbiology,2014,52(8):646-651.
[23]ISAACSON R,KIM H B.The intestinal microbiome of the pig[J].Animal Health Research Reviews,2012,13(1):100-109.
[24]KASUBUCHI M,HASEGAWA S,HIRAMATSU T,et al.Dietary gut microbial metabolites,short-chain fatty acids,and host metabolic regulation[J].Nutrients,2015,7(4):2839-2849.
[25]KOH A,DE VADDER F,KOVATCHEVA-DATCHARYP,et al.From dietary fiber to host physiology:shortchain fatty acids as key bacterial metabolites[J].Cell,2016,165(6):1332-1345.
[26]DUNCAN S H,HOLTROP G,LOBLEY G E,et al.Contribution of acetate to butyrate formation by human faecal bacteria[J].British Journal of Nutrition,2004,91(6):915-923.
[27]ZHANG Y J,LIU Q,ZHANG W M,et al.Gastrointestinal microbial diversity and short-chain fatty acid production in pigs fed different fibrous diets with or without cell wall-degrading enzyme supplementation[J].Livestock Science,2017,207:105-116.
[28]MEEHAN C J,BEIKO R G.A phylogenomic view of ecological specialization in the Lachnospiraceae,a family of digestive tract associated bacteria[J].Genome Biology and Evolution,2014,6(3):703-713.
[29]LOUIS P,HOLD G L,FLINT H J.The gut microbiota,bacterial metabolites and colorectal cancer[J].Nature Reviews Microbiology,2014,12(10):661-672.
[30]FLINT H J,BAYER E A,RINCON M T,et al.Polysaccharide utilization by gut bacteria:potential for new insights from genomic analysis[J].Nature Reviews Microbiology,2008,6(2):121-131.
[31]PARK J Y,LEE S H,KIM K R,et al.Production of13S-hydroxy-9(Z)-octadecenoic acid from linoleic acid by whole recombinant cells expressing linoleate 13-hydratase from Lactobacillus acidophilus[J].Journal of Biotechnology,2015,208:1-10.
[32]ORTEGA-ANAY J,HERNNDEZ-SANTOYO A.Production of bioactive conjugated linoleic acid by the multifunctional enolase from Lactobacillus plantarum[J].International Journal of Biological Macromolecules,2016,91:524-535.
[33]ZHANG D Y,SHANG T T,HUANG Y,et al.Gene expression profile changes in the jejunum of weaned piglets after oral administration of Lactobacillus or an antibiotic[J].Scientific Reports,2017,7:15816,doi:10.1038/s41598-017-16158-y.
[34]S'WIATKIEWICZ M,OCZKOWICZ M,ROPKA-MOLIKK,et al.The effect of dietary fatty acid composition on adipose tissue quality and expression of genes related to lipid metabolism in porcine livers[J].Animal Feed Science and Technology,2016,216:204-215.
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