微生态制剂调控比格幼犬生长性能和肠道菌群的研究

doi:10.16431/j.cnki.1671-7236.2022.02.012

摘要/Abstract

摘要： 【目的】研究复合微生态制剂对哺乳期比格幼犬生长性能与肠道菌群结构的影响。【方法】选取12只15日龄比格犬, 随机分为对照组和微生态制剂组, 每组6只。微生态制剂组幼犬每日灌喂犬用微生态制剂20 mL, 对照组幼犬灌喂生理盐水20 mL, 于试验第0、3、9、12、15、18、21、25、28 d测量体重并计算净增重, 第28天采集全血样、血清样和粪样。全血样进行红细胞(RBC)计数和血红蛋白(HGB)浓度测定, 血清样检测碱性磷酸酶(AKP)、无机磷(IP)、总胆固醇(TCHO)、甘油三酯(TG)、免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、白介素-1β(IL-1β)、脂多糖(LPS)含量; 粪样进行菌群测序, 统计Goods coverage指数、Observed species指数和Shannon指数, 对样本进行PCoA分析, 并统计各样本在门、属、种水平上的物种分布。【结果】与对照组相比, 微生态制剂可显著提高幼犬体重(P<0.05), 显著增加血清中RBC数量及HGB、AKP和IP含量(P<0.05), 促进比格幼犬生长发育; 微生态制剂可显著降低血清TG含量(P<0.05), 但对TCHO、IgA、IgG、IL-1β、LPS含量无显著影响(P>0.05)。Observed species指数和PCoA分析结果表明, 微生态制剂可显著改变幼犬肠道菌群结构(P<0.05)。在门水平上, 微生态制剂组幼犬肠道菌群拟杆菌门和放线菌门相对丰度显著降低(P<0.05), 梭菌门、酸杆菌门和疣微菌门相对丰度均显著增高(P<0.05);在属水平上, Escherichia-Shigella、霍尔德曼氏菌属、双歧杆菌属和布劳特氏菌属相对丰度均显著降低(P<0.05), 梭菌属、链球菌属、梭杆菌属、乳球菌属和片球菌属相对丰度均显著增高(P<0.05)。在种水平上, 大肠杆菌和短双歧杆菌相对丰度均显著降低(P<0.05), 犬大肠梭菌、格氏乳球菌、Vagococcus teuberi、贝莱斯芽孢杆菌、粪肠球菌和死亡梭杆菌相对丰度均显著增高(P<0.05)。【结论】犬用微生态制剂可显著促进幼犬生长, 提高其血氧含量, 改变肠道菌群结构, 优化肠道菌群组成。

关键词: 微生态制剂; 比格犬; 生长性能; 肠道菌群

Abstract: 【Objective】 This experiment was to explore the effects of compound probiotics on the growth performance and intestinal flora structure of Beagle puppies during lactation.【Method】 Twelve 15-day-old Beagle puppies were selected and randomly divided into control group and probiotics group, with 6 in each group.The puppies in the probiotics group were fed with 20 mL probiotic preparation daily, and the puppies in control group were fed with 20 mL normal saline.The body weight was measured and the weight gain was calculated on days 0, 3, 9, 12, 15, 18, 21, 25 and 28.Whole blood, serum and fecal samples were collected at the 28th day.Red blood cell count (RBC) and hemoglobin (HGB) concentrations were measured in whole blood samples, alkaline phosphatase (AKP), inorganic phosphorus (IP), total cholesterol (TCHO) content, triglyceride (TG), immunoglobulin A (IgA), immunoglobulin G (IgG), interleukin-1β (IL-1β), lipopolysaccharide (LPS) contents were measured in serum samples.The fecal samples were sequenced for flora, the Goods coverage index, Observed species index, and Shannon index were counted.The samples were analyzed by PCoA, and the species distribution at the phylum, genus and species levels of each sample were counted.【Result】 Compared with control group, probiotics could significantly increase the weight of puppies (P < 0.05), and significantly increase the number of RBC and the content of HGB, AKP and IP in serum (P < 0.05), and promote the growth and development of Beagle puppies.Probiotics could significantly reduce serum TG content (P < 0.05), but had no significant effect on TCHO, IgA, IgG, IL-1β and LPS (P < 0.05).Observed species index and PCoA analysis results showed that the probiotics could significantly change the structure of puppies intestinal flora (P < 0.05).At the phylum level, the relative abundance of Bacteroides and Actinobacteriota in the puppies of the microecological preparation group was significantly lower (P < 0.05), the relative abundance of Fusobacteriota, Acidobacteria and Verrucomicrobia were significantly increased (P < 0.05).At the genus level, the relative abundance of Escherichia-Shigella, Holdemanella, Bifidobacterium and Blautia was significantly reduced (P < 0.05), the relative abundance of Clostridium sensu stricto 1, Streptococcus, Fusobacteria, Lactococcus and Pediococcus was significantly increased (P < 0.05).At the species level, the relative abundance of Escherichia coli and Bifidobacterium breve was significantly decreased (P < 0.05), the relative abundance of Clostridium colicanis, Lactococcus garvieae, Vagococcus teuberi, Bacillus velezensis, Enterococcus faecium and Fusobacterium mortiferum was significantly increased (P < 0.05).【Conclusion】 The probiotics for puppies could significantly promote the growth of puppies, increase their blood oxygen content, change the structure of the intestinal flora, and optimize the composition of the intestinal flora.

Key words: probiotics; Beagle puppies; growth performance; intestinal flora

中图分类号:

S829.2

王志华, 贾俊鹏, RADEBE Stoffel Matjeke, 庾庆华. 微生态制剂调控比格幼犬生长性能和肠道菌群的研究[J]. 中国畜牧兽医, 2022, 49(2): 510-520.

WANG Zhihua, JIA Junpeng, RADEBE Stoffel Matjeke, YU Qinghua. Study on the Growth Performance and Intestinal Flora of Beagle Puppies Regulated by Probiotics[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 510-520.

参考文献

[1] CZARNECKI MAULDEN G. Effect of dietary modulation of intestinal microbiota on reproduction and early growth[J]. Theriogenology, 2008, 70(3): 286-290.
[2] SUN H Y, KIM K P, BAE C H, et al. Evaluation of Weissella cibaria JW15 probiotic derived from fermented korean vegetable product supplementation in diet on performance characteristics in adult beagle dog[J]. Animals, 2019, 9(8): 581.
[3] LEE H J, LEE S W. Effects of Lactobacillus acidophilus hy7032 and Lactobacillus reuteri hy7506 isolated from dog feces on welfare and hair in companion dogs[J]. Journal of Biomedical and Translational Research, 2021, 22(1): 32-41.
[4] XU H, ZHAO F, HOU Q, et al. Metagenomic analysis revealed beneficial effects of probiotics in improving the composition and function of the gut microbiota in dogs with diarrhoea[J]. Food & Function, 2019, 10(5): 2618-2629.
[5] 朱群, 常娟, 尹清强, 等. 益生菌对乳仔猪生长性能及肠道免疫和微生物区系的影响[A]. 中国畜牧兽医学会动物营养学分会第十二次动物营养学术研讨会[C]. 2016.
ZHU Q, CHANG J, YIN Q Q, et al. The effect of probiotics on the growth performance, intestinal immunity and microflora of suckling piglets[A]. The 12th Animal Nutrition Symposium of the Animal Nutrition Branch of the Chinese Society of Animal Husbandry and Veterinary Medicine[C]. 2016. (in Chinese)
[6] CUI C, SHEN C J, JIA G, et al. Effect of dietary Bacillus subtilis on proportion of Bacteroidetes and Firmicutes in swine intestine and lipid metabolism[J]. Genetics and Molecular Research, 2013, 12(2): 1766-1776.
[7] BIOURGE V, VALLET C L, LEVESQUE A, et al. The use of probiotics in the diet of dogs[J]. The Journal of Nutrition, 1998, 128(12): 2730S-2732S.
[8] 周顺伍. 动物生物化学[M]. 北京: 化学工业出版社, 2008.
ZHOU S W. Animal Biochemistry[M]. Beijing: Chemical Industry Press, 2008. (in Chinese)
[9] 李占占, 李勇, 丁雪, 等. 中药处方犬粮的安全性评价及其对犬创伤恢复能力的影响[J]. 安徽农业科学, 2014, 42(21): 7095-7098.
LI Z Z, LI Y, DING X, et al. Safety evaluation of Chinese medicine prescription dog food and its effect on dog's trauma recovery ability[J]. Journal of Anhui Agricultural Sciences, 2014, 42(21): 7095-7098. (in Chinese)
[10] SHARMA U, PAL D, PRASAD R. Alkaline phosphatase: An overview[J]. Indian Journal of Clinical Biochemistry, 2014, 29(3): 269-278.
[11] LAYDEN B T, ANGUEIRA A R, BRODSKY M, et al. Short chain fatty acids and their receptors: New metabolic targets[J]. Translational Research, 2013, 161(3): 131-140.
[12] MILLER T L, WOLIN M J. Pathways of acetate, propionate, and butyrate formation by the human fecal microbial flora[J]. Applied and Environmental Microbiology, 1996, 62(5): 1589-1592.
[13] LJUNGH A, WADSTROM T. Lactic acid bacteria as probiotics[J]. Current Issues in Intestinal Microbiology, 2006, 7(2): 73-90.
[14] BAILLON M L A, MARSHALL JONES Z V, BUTTERWICK R F. Effects of probiotic Lactobacillus acidophilus strain DSM13241 in healthy adult dogs[J]. American Journal of Veterinary Research, 2004, 65(3): 338-343.
[15] SILVA-MORITA F S D, BALBO S L, MENDES M C, et al. Short-chain fructooligosaccharides do not alter glucose homeostasis but improve the lipid profile in obese rats[J]. Acta Scientiarum Health Sciences, 2015, 37(2): 119-125.
[16] SEIFARTH C, HERING N A, ARNDT M, et al. Increased proinflammatory cytokines in mesenteric fat in major surgery and Crohn's disease[J]. Surgery, 2021, 169(6): 1328-1332.
[17] RAETZ C R, WHITFIELD C. Lipopolysaccharide endotoxins[J]. Annual Review of Biochemistry, 2002, 71(1): 635-700.
[18] YU H, GUO Z, SHEN S, et al. Effects of taurine on gut microbiota and metabolism in mice[J]. Amino Acids, 2016, 48(7): 1601-1617.
[19] 赵永玉. 微生态制剂对肉鸡生长性能、免疫功能、肠道菌群及血清生化指标的影响[J]. 饲料研究, 2021, 44(2): 50-53.
ZHAO Y Y. Effects of probioticss on growth performance, immune function, intestinal flora and serum biochemical indexes of broilers[J]. Feed Research, 2021, 44(2): 50-53. (in Chinese)
[20] MARTENS E C, NEUMANN M, DESAI M S. Interactions of commensal and pathogenic microorganisms with the intestinal mucosal barrier[J]. Nature Reviews Microbiology, 2018, 16(8): 457-470.
[21] ROSSI G, PENGO G, GALOSI L, et al. Effects of the probiotic mixture Slab51^{^®} (SivoMixx^{^®}) as food supplement in healthy dogs: Evaluation of fecal microbiota, clinical parameters and immune function[J]. Frontiers in Veterinary Science, 2020, 7: 613.
[22] PRENTICE A, EWING G, ROBERTS S B, et al. The nutritional role of breast-milk IgA and lactoferrin[J]. Acta Padiatrica, 1987, 76(4): 592-598.
[23] CABALLERO-FLORES G, SAKAMOTO K, ZENG M Y, et al. Maternal immunization confers protection to the offspring against an attaching and effacing pathogen through delivery of IgG in breast milk[J]. Cell Host & Microbe, 2019, 25(2): 313-323. e4.
[24] SHEN T D. Diet and gut microbiota in health and disease[J]. Nestle Nutrition Institute Workshop Series, 2017, 88: 117-126.
[25] 贾聪慧, 杨彩梅, 曾新福, 等. 丁酸梭菌对肉鸡生长性能、抗氧化能力、免疫功能和血清生化指标的影响[J]. 动物营养学报, 2016, 28(3): 908-915.
JIA C H, YANG C M, ZENG X F, et al. Effects of Clostridium butyricum on growth performance, antioxidant capacity, immune function and serum biochemical indexes of broilers[J]. Chinese Journal of Animal Nutrition, 2016, 28(3): 908-915. (in Chinese)
[26] CROXEN M A, FINLAY B B. Molecular mechanisms of Escherichia coli pathogenicity[J]. Nature Reviews Microbiology, 2010, 8(1): 26-38.
[27] PUJO J, PETITFILS C, LE FAOUDER P, et al. Bacteria-derived long chain fatty acid exhibits anti-inflammatory properties in colitis[J]. Gut, 2021, 70(6): 1088-1097.
[28] POYART C, QUESNE G, TRIEU-CUOT P. Taxonomic dissection of the Streptococcus bovis group by analysis of manganese-dependent superoxide dismutase gene (sodA) sequences: Reclassification of 'Streptococcus infantarius subsp. coli' as Streptococcus lutetiensis sp. nov. and of Streptococcus bovis biotype 11.2 as Streptococcus pasteurianus sp. nov[J]. International Journal of Systematic and Evolutionary Microbiology, 2002, 52(4): 1247-1255.
[29] 陈小云, 关孚时, 张存帅, 等. 产气荚膜梭菌主要外毒素最新研究进展[J]. 中国兽药杂志, 2005, 39(6): 29-33.
CHEN X Y, GUAN F S, ZHANG C S, et al. The latest research progress on the main exotoxins of Clostridium perfringens[J]. Chinese Journal of Veterinary Science, 2005, 39(6): 29-33. (in Chinese)
[30] SHEWMAKER P L, WHITNEY A M, GULVIK C A, et al. Vagococcus bubulae sp. nov., isolated from ground beef, and Vagococcus vulneris sp. nov., isolated from a human foot wound[J]. International Journal of Systematic and Evolutionary Microbiology, 2019, 69(8): 2268-2276.
[31] TORRES M, LLAMAS I, TORRES B, et al. Growth promotion on horticultural crops and antifungal activity of Bacillus velezensis XT1[J]. Applied Soil Ecology, 2020, 150: 103453.
[32] HOLZAPFEL W, ARINI A, AESCHBACHER M, et al. Enterococcus faecium SF68 as a model for efficacy and safety evaluation of pharmaceutical probiotics[J]. Beneficial Microbes, 2018, 9(3): 375-388.
[33] JUNGERSEN M, WIND A, JOHANSEN E, et al. The science behind the probiotic strain Bifidobacterium animalis subsp. lactis BB-12[J]. Microorganisms, 2014, 2(2): 92-110.