Method for separating, identifying and preliminary screening of probiotic yak-derived lactic acid bacteria
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A technology of probiotics and lactic acid bacteria, applied in the biological field, can solve problems such as irrational use of antibiotics and multi-drug resistance, and achieve the effects of protecting intestinal mucosa, reducing inflammation and diarrhea, and regulating immunity
Inactive Publication Date: 2018-08-24
TIBET AGRI & ANIMAL HUSBANDRY COLLEGE
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Due to the irrational use of antibiotics in the treatment of the disease, th
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Embodiment 1
[0021] A method for the separation, identification and preliminary screening of probiotic yak-derived lactic acid bacteria of the present invention comprises the following steps:
[0022] 1 Materials and methods
[0023] 1.1 Sample collection
[0024] After slaughtering a 2.5-year-old healthy yak in Nyingchi, Tibet, its duodenum, jejunum, ileum, cecum, colon, rectum and their contents were aseptically collected, put into a sterilized plate, and then put into a refrigerated anaerobic box , and bring it back to the lab as soon as possible.
[0025] 1.2 Reagents and equipment
[0026] MRS agar, MRS broth, and MacConkey agar medium are products of Qingdao High-tech Park Haibo Biotechnology Co., Ltd.; blood agar medium is purchased from Chengdu Jierui Technology Co., Ltd.; pig bile salt is a product of Biotopped; hydrochloric acid and glycerin are products of Sinopharm Products of Group Chemical Trial Manufacturing Co., Ltd. Bacterial Genome DNA Extraction Kit was purchased fro...
Embodiment 2
[0074] A method for the separation, identification and preliminary screening of probiotic yak-derived lactic acid bacteria of the present invention comprises the following steps:
[0075] 1 Materials and methods
[0076] 1.1 Sample collection
[0077] After slaughtering a 2.5-year-old healthy yak in Nyingchi, Tibet, its duodenum, jejunum, ileum, cecum, colon, rectum and their contents were aseptically collected, put into a sterilized plate, and then put into a refrigerated anaerobic box , and bring it back to the lab as soon as possible.
[0078] 1.2 Reagents and equipment
[0079] MRS agar, MRS broth, and MacConkey agar medium are products of Qingdao High-tech Park Haibo Biotechnology Co., Ltd.; blood agar medium is purchased from Chengdu Jierui Technology Co., Ltd.; pig bile salt is a product of Biotopped; hydrochloric acid and glycerin are products of Sinopharm Products of Group Chemical Trial Manufacturing Co., Ltd. Bacterial Genome DNA Extraction Kit was purchased fro...
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Abstract
The invention belongs to the field of biotechnology, and discloses a method for separating, identifying and preliminary screening of probiotic yak-derived lactic acid bacteria. The separation steps comprise: taking about 1 g of the middle portion of a manure sample, adding into a sterilized and refrigerated peptone buffer solution, sucking 1 mL of the manure supernatant under a sterile condition,inoculating into a MRS liquid culture medium, respectively carrying out aerobic culture and anaerobic culture for 24 h at a temperature of 37 DEG C, taking 200 [mu]l of the bacterial suspension, coating a MRS plate with the bacterial suspension, respectively carrying out aerobic culture and anaerobic culture for 24 h, selecting single suspected colony, inoculating into a corresponding plate culture medium, carrying out streak culture, respectively carrying out aerobic culture and anaerobic culture for 24 h at a temperature of 37 DEG C, re-selecting single colony, inoculating into a corresponding plate culture medium, respectively carrying out aerobic culture and anaerobic culture at a temperature of 37 DEG C, purifying twice, selecting single colony, inoculating into a corresponding slopeculture medium, respectively carrying out aerobic culture and anaerobic culture for 24 h at a temperature of 37 DEG C, and storing in a 4 DEG C a refrigerator so as to be spare. According to the present invention, the probiotic yak-derived lactic acid bacteria can protect the intestinal mucosa so as to reduce intestinal epithelium inflammation and diarrhea.
Description
technical field [0001] The invention specifically relates to a method for the isolation, identification and preliminary screening of probiotic yak-derived lactic acid bacteria, which belongs to the field of biotechnology. Background technique [0002] Calf diarrhea disease has always been an important disease affecting the yak breeding industry in Tibet. Its causes are complex and mostly occur in the alternate seasons of winter and spring. It occurs in all counties of Tibet. According to the survey, in Bangor County, among the calves that died in spring Diarrhea killed up to 70%. Due to the irrational use of antibiotics in the treatment of the disease, there are serious multi-drug resistances in many pathogenic bacteria in yaks. Therefore, there is an urgent need to find effective and low-risk methods for the treatment and prevention of diarrheal disease in calves. [0003] Probiotics are live microorganisms that are beneficial to the health of the host when ingested in su...
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