鸡源凝结芽孢杆菌的分离鉴定及耐受性分析

doi:10.16431/j.cnki.1671-7236.2022.11.009

Abstract

Abstract: 【Objective】 The aim of this experiment was to find a Bacillus coagulans strain with stable performance and strong environmental resistance, so as to play its role instead of antibiotics in actual production.【Method】 The fecal samples from 160-day old healthy Hyline Brown laying hens were collected for isolation and purification of bacteria, and morphological characteristics observation, physiological and biochemical identification, 16S rDNA sequencing analysis and identification of the isolated bacteria were carried out.The growth rule was determined by plotting the growth dynamic curve of the bacterium.Acid and bile salt resistance tests were used to analyze the resistance of the strain.The culture conditions of isolated strains were optimized by single factor experiment.【Result】 The isolated bacteria grew gray white colonies with rough and irregular surface on the ordinary culture medium, and were rod-shaped and spore terminal Gram-positive bacteria under microscope.Combined with biochemical experiments and 16S rDNA sequence analysis, the strain was identified as Bacillus coagulans, named NJ001.The growth dynamic curve showed that 7-24 h was the logarithmic growth period of the strain, and the number of viable number reached the peak at 24 h.The optimal culture time of Bacillus coagulans NJ001 was 24 h.The results of tolerance test showed that the survival rate was 59.5% at pH 3.0 for 2 h, and 87.6% at pH 4.0, showing strong resistance to acid.The survival rate was 54.0% after treatment with 0.3% bile salt for 2 h, and the survival rate was still more than 50% when the concentration of bile salt increased to 0.5%.The results of single factor experiment showed that the optimum culture temperature was 40 ℃, inoculation amount 2%, initial pH 7.0 and shaking speed was 220 r/min.Based on the above optimum cultivation conditions, the maximum viable count of the strain was 1.93×10⁸ CFU/mL.【Conclusion】 The isolated strain was Bacillus coagulans with strong acid and bile salt resistance, and its optimal growth conditions which was basically consistent with the gastrointestinal environment of animals were easy to achieve, so it had the potential of application as a microecological agent.

Key words: chicken; Bacillus coagulans; isolation and identification; stress reistance

CLC Number:

S852.61⁺6

SUN Yanting, KANG Yu, ZHAN Yan, ZHANG Dongxuan, LIU Chang, BIAN Chuanzhou, ZHANG Xiaojing, BAI Jing, QIAO Hongxing. Isolation,Identification and Stress Resistance Analysis of Bacillus coagulans from Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4187-4196.

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Isolation,Identification and Stress Resistance Analysis of Bacillus coagulans from Chickens

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