A kind of lactobacillus and its application for inhibiting pathogenic Escherichia coli of avian origin
A technology of Lactobacillus and colibacillosis, which is applied in the direction of bacteria, antibacterial drugs, and methods based on microorganisms, can solve the problems of poultry farming, such as economic harm, chick death, and rough feathers, so as to protect food safety and human health , Inhibition of adhesion, good stability
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Embodiment 1
[0026] 1. The source of Lactobacillus SCGZ-9
[0027] The Lactobacillus was isolated by the inventor from healthy chickens in a Guangdong farm in mid-2013, and is now preserved and provided by the Poultry Disease Research Office of the Veterinary College of South China Agricultural University.
[0028] 2. Identification and classification of Lactobacillus SCGZ-9
[0029] (1) Biochemical identification
[0030] According to the "Berger's Bacterial Identification Manual", do the following biochemical tests: catalase test, nitrate reduction test, gelatin liquefaction test, indole test, hydrogen sulfide test, V-P test and dynamic test. The seven biochemical tests were all negative. It was determined that the screened SCGZ-9 strain belonged to the genus Lactobacillus.
[0031] (2) 16S rRNA gene sequencing
[0032] Design primer sequences: fD1: 5'-AGAGTTTGATCCTGGCTCAG-3', rD1: 5'-AAGGAGGTGATCCAGCC-3', PCR reaction was carried out according to the instruction manual of rTaq DNA p...
Embodiment 2
[0044] 1. In vivo environmental tolerance test
[0045] The digestive tract of the body is a complex environment, especially the stomach and intestinal tract. The environment in the stomach of chickens is strongly acidic, which can kill most of the microorganisms that enter the stomach, while there are bile and various secretions in the small intestine, which can inhibit microbial growth. For probiotics to enter the intestinal tract and play a role, in addition to providing protection through the production process strains, the strains themselves should also have a certain tolerance to environments such as low pH and the presence of bile salts. Many current studies use the method of simulating the gastrointestinal environment in vitro to evaluate the tolerance of probiotics to the in vivo environment.
[0046] (1) Acid resistance test
[0047] The SCGZ-9 strain was passaged three times in MRS broth, and the concentration was adjusted at 10 8 About CFU / mL, inoculate 5% (v / v)...
Embodiment 3
[0055] Example 3: Inhibitory effect of Lactobacillus SCGZ-9 on pathogenic Escherichia coli in vivo
[0056] 1. Agar Diffusion Test
[0057] The SCGZ-9 strain was inoculated in MRS broth, recovered at 37°C, subcultured twice after 24 hours, inoculated in MRS broth, centrifuged at 8000r / min for 20min, and the supernatant was taken, which was the suspension of Lactobacillus SCGZ-9. liquid.
[0058]Pathogenic Escherichia coli (Enteropathogenic Escherichia coli, EPEC) (chicken pathogenic Escherichia coli O 78 , serotype O78: K80, number GIM1.739, purchased from Guangdong Microbial Culture Collection Center) densely spread on LB nutrient agar plate (ingredients (g / L): tryptone 10.0, yeast extract powder 5.0, sodium chloride 10.0, agar 15.0, pH value 7.0±0.2), make chicken pathogenic Escherichia coli O 78 Cover the plate evenly and place it at 37°C for 20min. Take out the flat plate in the ultra-clean bench, and punch three holes vertically on the flat plate with a sterile punche...
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