一株鸡源枯草芽孢杆菌的分离鉴定及其对雏鸡腹泻的保护作用

doi:10.11843/j.issn.0366-6964.2025.12.037

摘要/Abstract

摘要：

旨在从健康蛋鸡粪便样品中分离鉴定枯草芽孢杆菌(Bacillus subtilis, B.S)，并探究其对大肠杆菌(Escherichia coli, E.coli)诱导蛋雏鸡腹泻的保护作用。试验采用形态鉴定和16S rDNA测序法分离、筛选鉴定B.S，检测其对pH、胆盐和人工胃肠液的耐受性及溶血性。动物试验选取96只1日龄健康大午金凤蛋鸡，随机分为8组，每组12只鸡，分别为对照组(饲喂基础日粮，CON)、低剂量组(饲喂基础日粮的基础上，口服1 mL×10⁹ CFU的B.S JZH 9，LBS)、中剂量组(饲喂基础日粮的基础上，口服1 mL×10¹⁰ CFU的B.S JZH 9，MBS)、高剂量组(饲喂基础日粮的基础上，口服1 mL×10¹¹ CFU的B.S JZH 9，HBS)、模型组(饲喂基础日粮的基础上，口服1 mL×4.5×10¹² CFU的E.coli O78，Model)、低剂量预防组(LBS + E.coli)、中剂量预防组(MBS + E.coli)、高剂量预防组(HBS + E.coli)，研究口服B.S JZH 9对健康及E.coli O78攻毒下蛋雏鸡生长性能和肠道健康的影响，试验周期为21 d，1~18 d口服B.S JZH 9，19~21 d口服E.coli O78。结果表明：分离菌株为革兰阳性菌，结合16S rDNA鉴定确定为B.S，命名为B.S JZH 9，耐酸、耐胆盐，无溶血反应，对人工胃肠液具有耐受性。动物试验结果表明：与CON相比，无攻毒的LBS、MBS、HBS对1~21 d的平均日增重(average daily gain, ADG)、平均日采食量(average daily feed intake, ADFI)、饲料转化率(feed conversion rate, FCR)无显著影响；与Model相比，LBS + E.coli有降低1~21 d FCR的趋势；LBS + E.coli、MBS + E.coli、HBS + E.coli显著提高了十二指肠绒毛高度和绒隐比，MBS + E.coli组、HBS + E.coli组显著降低了十二指肠隐窝深度；与CON相比，Model空肠E.coli含量显著升高，饲喂不同浓度的B.S JZH 9不同程度地降低了E.coli的相对含量。综上所述，源自健康蛋鸡粪便的B.S JZH 9，通过促进雏鸡生长性能，增加小肠绒毛高度和绒隐比，降低隐窝深度，增大小肠的吸收面积，降低空肠E.coli的含量，从而调节肠道健康，减缓大肠杆菌对肠道造成的损伤。

关键词: 枯草芽孢杆菌, 蛋鸡, 生长性能, 肠道健康

Abstract:

This experiment mainly carried on isolation and identification Bacillus subtilis (B.S) from healthy egg chicken fecal samples, and explored the protective effect on E.coli O78 induced diarrhea in chicks. The experiment used morphological identification and 16S rDNA sequencing to isolate, screen and identify B.S. The tolerance to pH, bile salts, and artificial gastrointestinal fluids and hemolysis was assessed. Ninty-six healthy 1-day-old Dawu Jinfeng laying hens were selected for animal experiments and randomly divided into eight groups, 12 chickens per group: The control group (fed with basal diet, CON), low dose group (fed with basal diet and orally administeres 1 mL×10⁹ CFU of B.S JZH 9, LBS), medium dose group (fed with basal diet and orally administeres 1 mL×10¹⁰ CFU of B.S JZH 9, MBS), and high dose group (fed with basal diet and orally administeres 1 mL×10¹¹ CFU of B.S JZH 9, HBS). The model group (fed with basal diet and orally administered 1 mL×4.5×10¹² CFU of E.coli O78, Model), low dose prevention group (LBS + E.coli), medium dose prevention group (MBS + E.coli), and high dose prevention group (HBS + E.coli). The study analyzed the effects of the B.S JZH 9 on the growth performance and intestinal health of E.coli O78 challenged laying chicks. The experiment lasted for 21 days, with oral administration of B.S JZH 9 fod 1~18 days and oral administration of E.coli O78 for 19~21 days. The results showed that the isolated Gram-positive bacteria, identified as B.S by 16S rDNA, named B.S JZH 9. With the characteristics of acid and bile salt resistance, no hemolytic reaction, and tolerance to artificial gastrointestinal fluids. The results of animal testing showed that compared with the CON, the LBS, MBS, HBS had no significant effect on average daily gain (ADG), average daily feed intake (ADFI), and feed conversion rate (FCR) from 1 to 21 days. Compared with the Model, the LBS + E.coli showed a trend of reducing FCR from 1 to 21 days and the LBS + E.coli, MBS + E.coli and HBS + E.coli significantly increased the villus height and villus-crypt ratio, while the MBS + E.coli and HBS + E.coli significantly reduced the crypt depth of duodenal. Compared with the CON, the content of E.coli was significantly increased in the jejunum of the Model, while feed different concentrations of B.S JZH 9 reduced the relative content of E.coli to varying degrees. In summary, B.S JZH 9 isolated from the feces of healthy laying hens, promoted growth performance, increased the villus height and villus-crypt ratio of small intestinal, reduced crypt depth, increased the absorption area of the small intestine, and reduced the content of E.coli in the jejunum, thereby regulating intestinal health and slowing down the damage caused by E.coli O78 to the intestine.

Key words: Bacillus subtilis, laying hen, growth properties, intestinal health

中图分类号:

S852.6

吕紫一, 张新博, 王红志, 郭宏磊, 赵春旭, 刘顺波, 马宁. 一株鸡源枯草芽孢杆菌的分离鉴定及其对雏鸡腹泻的保护作用[J]. 畜牧兽医学报, 2025, 56(12): 6351-6363.

LÜ Ziyi, ZHANG Xinbo, WANG Hongzhi, GUO Honglei, ZHAO Chunxu, LIU Shunbo, MA Ning. Isolation and Identification of a Chicken Source Bacillus subtilis Strain and Its Protective Effect on Diarrhea in Chicks[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6351-6363.

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组别Group	1~18 d		19~21 d
组别Group	B.S JZH 9	E.coli O78	B.S JZH 9	E.coli O78
对照组CON	/	/	/	/
低剂量组LBS	1 mL×10⁹ CFU·d^-1	/	/	/
中剂量组MBS	1 mL×10¹⁰ CFU·d^-1	/	/	/
高剂量组HBS	1 mL×10¹¹ CFU·d^-1	/	/	/
模型组Model	/	/	/	1 mL×4.5×10¹² CFU·d^-1
低剂量预防组LBS + E.coli	1 mL×10⁹ CFU·d^-1	/	/	1 mL×4.5×10¹² CFU·d^-1
中剂量预防组MBS + E.coli	1 mL×10¹⁰ CFU·d^-1	/	/	1 mL×4.5×10¹² CFU·d^-1
高剂量预防组HBS + E.coli	1 mL×10¹¹ CFU·d^-1	/	/	1 mL×4.5×10¹² CFU·d^-1

项目 Item	初始体重/g Initial weight	末重/g Final weight	平均日增重/(g·d^-1) ADG	平均日采食量/(g·d^-1) ADFI	饲料转化率(F/G) FCR
CON	77.3±9.5	329±28.6	12.0±1.2^a	25.5±7.3	2.1±0.21^a
LBS	75.0±6.7	309±28.1	11.6±1.1^ab	22.2±6.6	2.0±0.21^a
MBS	76.2±6.0	311±21.3	11.2±0.9^ab	23.5±6.4	2.1±0.16^a
HBS	74.3±7.1	306±41.3	11.0±1.8^ab	22.5±5.5	2.1±0.35^a
Model	78.7±6.9	311±34.3	11.1±1.6^ab	24.2±6.2	2.2±0.37^ab
LBS + E.coli	77.4±6.4	315±21.2	11.4±0.9^ab	24.0±5.7	2.1±0.15^a
MBS + E.coli	74.3±7.9	282±52.0	9.7±2.6^c	22.5±4.3	2.5±0.55^b
HBS + E.coli	76.0±5.7	292±27.3	10.3±1.1^bc	22.9±5.0	2.2±0.23^ab

组别 Group	绒毛高度/μm Villus height	隐窝深度/μm Crypt depth	绒隐比Villus to crypt depth ratio
CON	1 193±58.2^b	108±34.4^c	11.7±3.13^a
LBS	1 001±49.0^c	122±20.6^bc	8.36±1.19^b
MBS	1 223±103^ab	125±21.2^bc	9.90±1.61^ab
HBS	1 355±144^a	145±18.2^ab	9.34±0.40^ab
Model	874±54.8^d	169±16.6^a	5.16±0.18^c
LBS + E.coli	1 239±30.0^ab	146±7.7^ab	8.48±0.33^b
MBS + E.coli	1 198±21.5^b	134±10.2^bc	8.96±0.73^b
HBS + E.coli	1 301±33.3^ab	121±8.4^bc	10.8±0.63^ab

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