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Screening and application of probiotic Enterococcus faecium

A technology of Enterococcus faecium and Staphylococcus, applied in the application, microorganism-based methods, bacteria and other directions, can solve problems such as harming the healthy development of aquaculture, affecting human food safety, and normal flora imbalance in animals, so as to improve livestock and poultry production. Effects of performance, human and animal safety, production and ease of use

Active Publication Date: 2012-10-24
武汉华大瑞尔科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the extensive use of antibiotics, especially unscientific abuse, problems such as the emergence of a large number of drug-resistant strains, the imbalance of normal animal flora, and drug residues in animal products have become increasingly prominent, seriously endangering the healthy development of the aquaculture industry and affecting human food safety. , has aroused the great attention of governments and industry insiders all over the world

Method used

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  • Screening and application of probiotic Enterococcus faecium
  • Screening and application of probiotic Enterococcus faecium
  • Screening and application of probiotic Enterococcus faecium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Isolation and identification of probiotic strains

[0047] 1. Isolation of strains

[0048] The samples were taken from the 100-day-old and 21-day-old Chinese local pig breed Hubei "Tongcheng Pig" (the breed comes from the breeding farm of Tongcheng County, Hubei Province), and the rectal contents of the pigs were collected from the anus with sterilized cotton swabs. After the sample is collected, use a cotton swab to streak and culture it in KF Streptococcus agar medium (purchased from Hangzhou Microbial Reagent Co., Ltd.). Colonies of large size were purely cultured. The pure culture was preliminarily screened by colony morphology observation, Gram staining and peroxidase test. Bacteria in the form of single, paired or short-chain oval Gram-positive cocci and strains negative for peroxidase tests are all passaged and preserved. The growth characteristic of the bacterial strain of this invention on the KF Streptococcus agar medium sees figure 1 , for its...

Embodiment 2

[0089] Embodiment 2: Bacteriostatic performance test of Enterococcus faecium strain HDRsEf1

[0090] Staphylococcus aureus (ATCC25923, purchased from the Clinical Inspection Center of the Ministry of Health), Salmonella choleraesuis C78-1 (purchased from China Veterinary Drug Administration) and porcine pathogenic Escherichia coli O138:K88 (C83902, purchased from China Supervision Institute) is indicator bacterium, with HDRsEf1 bacterial strain fermentation supernatant of the present invention is antibacterial agent, detects the in vitro bacteriostatic performance of Enterococcus faecium bacterial strain HDRsEf1, concrete operation is as follows:

[0091] 1) Preparation and treatment of bacterial fermentation broth: Inoculate fresh Enterococcus faecium HDRsEf1 bacterial fluid into MRS broth medium (product of BD Company, USA) at 2% (v / v), culture it statically for 18-24 hours, and take it by centrifugation. The supernatant was centrifuged at 10000rpm for 30min, and the supernata...

Embodiment 3

[0100] Example 3: Safety evaluation of Enterococcus faecium strains

[0101] 1. Amplification of virulence factors

[0102] The amplification of virulence factors mainly includes cylA (accession number: AAA62652.1; Gilmore et al., 1994), gelE (accession number: D85392; Mannu et al., 2003), esp (accession number: AF034779; Shankar et al. ., 1999), agg (no accession number: Galli et al., 1990), ace (accession number: AF26083; Mannu et al., 2003), efaAfs (accession number: EFU03756) and efaA fm (Accession number: FJ609170.1; Mannu et al., 2003).

[0103] Using the Enterococcus genome prepared above as a template, PCR amplification was performed with specific primer pairs for each virulence factor. Each primer was synthesized by Shanghai Yingjun Biotechnology Co., Ltd., its sequence, PCR product size, amplification conditions and references See Table 4, and see Table 5 for the virulence gene PCR amplification system.

[0104] Table 4 Test primer sequences, PCR product size and ...

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Abstract

The invention belongs to the technical field of veterinary microbial additive preparation, and specifically relates to a strain of separated and screened Enterococcus faecium providing significant bacterial inhibition effects for common enteropathogenic bacteria such as staphylococcus aureus, escherichia coli and salmonella in breeding animals, and an application thereof. The probiotic Enterococcus faecium of the present invention is characterized in that: the strain is Enterococcus faecium HDRsEf1, and is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CCTCC NO:M2011031. The probiotic Enterococcus faecium of the present invention has characteristics of fast growth, high acid production capacity, strong stress resistance, safety, disease resistance and growth promotion, and can be used as the microbial feed additive for livestock and poultry feeds.

Description

technical field [0001] The invention belongs to the technical field of veterinary microbial additives and is related to the field of antibiotic additives. The invention relates to an Enterococcus faecium (Enterococcus faecium) which has obvious antibacterial effects on common intestinal pathogenic bacteria such as Staphylococcus aureus, Escherichia coli and Salmonella. faecium) strain isolation and identification, safety evaluation, stress resistance performance and probiotic performance identification and use as a feed additive. Background technique [0002] Since Moore first reported in 1946 that adding antibiotics to feed can significantly increase the daily weight gain of broilers, more than 60 kinds of antibiotics have been used in animal husbandry, playing an important role in the prevention and control of animal diseases and improving the performance of livestock and poultry. However, with the extensive use of antibiotics, especially unscientific abuse, problems such ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K1/16A61P31/04C12R1/01
Inventor 石德时肖运才毕丁仁熊嫒嫒王喜亮高晓聪许光胜李自力周祖涛刘梅许青荣郝晓莉肖宏德
Owner 武汉华大瑞尔科技有限公司
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