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Novel enterococcus and streptococcus strains and bacteriocins

A technology of bacteriocin and streptococci, applied in inducer strains of Lactococcus, diseases in poultry, application fields of therapeutic compositions, can solve problems such as difficult adjustment of immune response

Inactive Publication Date: 2009-05-06
THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SEC OF AGRI (US) +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immune responses in the gastrointestinal (GI) tract are not easily regulated

Method used

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  • Novel enterococcus and streptococcus strains and bacteriocins
  • Novel enterococcus and streptococcus strains and bacteriocins
  • Novel enterococcus and streptococcus strains and bacteriocins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Two novel antagonistic strains, Enterococcus faecium strain 50-52 (NRRL B-30746) and Streptococcus hamster strain 760 (NRRL B-30745), isolated from the mucosal surface of caecum of approximately 1.0 g of broiler chickens, were suspended in Add about 10 ml of sterile 0.85% (w / v) saline solution (physiological saline) and heat to about 80°C for about 15 minutes. About 0.10 ml of the about 1:50 and 1:2500 suspensions were spread onto plate count agar or MRS agar. The plates were incubated at about 37°C for about 24 hours under microaerobic conditions. Colonies with different morphologies were streaked on MRS agar. These cultures were incubated at about 37°C for about 24 hours under microaerobic conditions.

[0057] Strain 760 was grown on MRS agar at about 37°C for about 24 hours. The strains are facultative aerobes and Gram-positive cocci which can grow between about 37°C and 45°C. The strain grows on nutrient agar or plate count agar and after about 24 hours of aerob...

Embodiment 2

[0060] Crude antimicrobial preparations were extracted from cultures of two antagonistic strains: Enterococcus 50-52 and Streptococcus 760. The antagonist was incubated aerobically with the inducer strain L. lactis strain 320 (NRRL B-30744, supra) in about 250 ml of 10% Brucella broth for about 14 hours at 37°C. The concentration of the producer strain was about 106 CFU / mL and the inducer strain was about 10 7 CFU / mL. The resulting culture was centrifuged at about 2,500 xg for about 10 minutes to remove most of the viable cells. The decanted supernatant was mixed with about 60% saturated ammonium sulfate and incubated at about 4°C for about 24 hours to precipitate the bacteriocin compounds. After centrifugation at about 10,000 xg for about 20 minutes, the pellet was resuspended in about 1.5 ml of about 10 mM sodium phosphate buffer, pH about 7.0, and dialyzed overnight against about 2.5 L of the same buffer. The resulting solution was referred to as crude antimicrobial prep...

Embodiment 3

[0061] The antimicrobial spectrum of the CAPs was determined using a spot test. About 1 ml of the sterilized crude antimicrobial preparation (CAP) obtained in Example 2 above was diluted with about 1 ml of phosphate-sodium salt buffer (pH about 7.0) and sterilized as in Example 2. About 10 microliters of each sample was plated onto blood-supplemented Campylobacter agar or nutrient agar (MPA or Meta-Peptone Agar) pre-inoculated with cells of the bacteria of interest. Plates containing Campylobacter jejuni were grown under microaerobic conditions at about 42°C, and Y.enterocolitica and Y.pseudotuberculosis were cultured aerobically at about 28°C. ), other bacterial strains were incubated aerobically at about 37°C for about 24 or 48 hours. Identification is based on regions of inhibition produced by the target bacteria. CAP activity is expressed in arbitrary units (AU) per milliliter of preparation where a visible region of culture growth inhibition occurs (Henderson et al., Ar...

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Abstract

Novel Enterococcus and Streptococcus bacteriocins produced by novel Enterococcus and Streptococcus strains are used for at least reducing the levels of colonization by at least one target bacteria in animals, especially poultry.

Description

Background of the invention [0001] field of invention [0001] The present invention relates to the control of disease in animals, especially poultry, by the use of novel bacteriocin-producing Enterococcus and Streptococcus strains and / or novel bacteriocins produced by these strains. The present invention also relates to a new bacteriocin, the amino acid sequence of said new bacteriocin, and to an Enterococcus or Streptococcus producing said new bacteriocin, and to an inducer strain of Lactococcus. Further, the present invention relates to therapeutic compositions containing said novel bacteriocins and / or Enterococci or Streptococci producing them, and uses of said therapeutic compositions. Description of related fields [0002] Intestinal disease has been caused in humans by consumption of improperly handled poultry products. Salmonella spp. has long been recognized as the causative bacterium for such diseases, and recently Campylobacter spp., especially Campylobacter ...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61K38/17C07K14/335C07K14/195C12N1/20
CPCC07K14/315C12R1/46A61K38/00A23K1/1631A23K20/147C12R2001/46C12N1/205
Inventor N·斯特恩J·莱恩E·斯韦托奇B·叶鲁斯拉诺夫V·佩雷列金E·米特西韦奇I·米特西韦奇V·列夫查克
Owner THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SEC OF AGRI (US)
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