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Detection of antibacterial activity in excretory secretory product of adult trichuris suis

a technology of excretory secretory products and antibacterial activity, which is applied in the direction of biocide, peptide/protein ingredients, peptide sources, etc., can solve the problems that many antibiotics have been rendered ineffective in preventing bacterial infections

Inactive Publication Date: 2006-09-28
BOARD OF TRUSTEES OPERATING MICHIGAN STATE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides an antibacterial activity from the excretory-secretory products of Trichuris suis that is resistant to boiling, freeze-thawing, trypsin, and pronase E. The activity has been found to inhibit the growth of various bacteria, including Escherichia coli, Campylobacter jejuni, and Staphylococcus aureus. The excretory-secretory product can be used in applications for killing or inhibiting the growth of microorganisms. The invention also provides a method for isolating the antibacterial activity from the excretory-secretory products of Trichuris suis. The isolated antibacterial activity has a minimum inhibitory concentration and a minimum bactericide concentration of about the same. The excretory-secretory product is effective in inhibiting at least 50% of at least one species of cream positive or cream negative bacteria.

Problems solved by technology

However, because many antibiotics have been used with relative abandon over the past half century, many microorganisms have developed resistance to many of the antibiotics.
As a consequence, many antibiotics have been rendered ineffective at preventing bacterial infections.

Method used

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  • Detection of antibacterial activity in excretory secretory product of adult trichuris suis
  • Detection of antibacterial activity in excretory secretory product of adult trichuris suis
  • Detection of antibacterial activity in excretory secretory product of adult trichuris suis

Examples

Experimental program
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Effect test

example 1

[0053] This example illustrates the discovery and subsequent isolation of the novel trichuricin from the ESP of Trichuris suis and further provides a preliminary analysis of the novel trichuricin.

Materials and Methods

[0054] Experimental infection of pigs and recovery of adult Trichuris suis. Weaned pigs, were maintained in confinement housing and provided pig chow and water ad libitum. The pigs were experimentally infected with approximately 2500 viable Trichuris suis eggs by oral gavage. Pigs were killed 45 to 50 days after infection with a captive bolt gun. The necropsy procedure involved opening the abdomen and removing the entire GI tract. The GI tract was slit open longitudinally, contents emptied into a bucket, and adult whipworms plucked from the colon with forceps. Intestinal contents were washed through a 2 mm sieve with tap water and additional whipworms were recovered from the sieve. Whipworms were picked individually using forceps into successive petri dishes of steril...

example 2

[0087] This example shows the sequencing and synthesis of trichuricin.

[0088] The trichuricin is prepared and isolated from the ESP as in Example 1. The trichuricin is sequenced using peptide sequencing methods well known in the art. The sequence of the trichuricin antibacterial activities enables degenerate PCR primers to be made which can be used to identify the Trichuris suis genomic DNA encoding the antibacterial activities. The amino acid sequence allows synthetic trichuricin to be chemically synthesized using peptide synthesis technologies well known in the art.

example 3

[0089] This example shows the preparation of monoclonal antibodies that recognize the trichuricin.

[0090] The trichuricin is prepared and isolated from the ESP as in Example 1 or synthesized as in Example 2. Then the trichuricin is used to make monoclonal antibodies according to the methods in Antibodies, A Laboratory Manual. Harlow and Lane (Eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1988), well known to those skilled in the art as a source for methods for making polyclonal and monoclonal antibodies. Because of the small size of the trichuricin (less than or equal to 100 amino acids), the trichuricin is preferably coupled to a carrier such as bovine serum albumin or keyhole limpet hemocyanin with a bifunctional reagent such as glutaraldehyde (amino to amino), m-Maleimidobenzoic acid-N-hydroxy-succinimide (MBS; amino to sulfhydryl), bisdiazo-benzidine (BDB; tyr to tyr), or carbodiimide (EDAC; amino to carboxyl) using methods well known in the art.

[0091] B...

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Abstract

The present invention provides a heat-stable and protease-resistant antibacterial activity in excretory-secretory products (ESP) of Trichuris suis. The antibacterial activity is not more than 10,000 MW; is resistant to boiling, trypsin, and pronase E; has a bactericidal mode of action; and is effective against Gram positive and Gram negative bacteria, including Escherichia coli, Campylobacter jejuni, Campylobacter coli, and Staphylococcus aureus. The antibacterial activity is useful in applications for killing or inhibiting the growth of microorganisms, in particular bacteria.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application claims priority to Provisional Application No. 60 / 246,203, which was filed Nov. 6, 2000.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT [0002] This invention was supported by National Institutes of Health Grant No. AI42348-03. The U.S. government has certain rights in this invention.REFERENCE TO A “COMPUTER LISTING APPENDIX SUBMITTED ON A COMPACT DISC”[0003] Not Applicable. BACKGROUND OF THE INVENTION [0004] (1) Field of the Invention [0005] The present invention relates to heat-stable and protease-resistant antibacterial activity in excretory-secretory products (ESP) of Trichuris suis. The antibacterial activity is not more than 10,000 MW; is resistant to boiling, trypsin, and pronase E; has a bactericidal mode of action; and is effective against Gram positive and Gram negative bacteria, including Escherichia coli, Campylobacter jejuni, Campylobacter coli, and Staphylococcus aureus. The antibacterial act...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01K67/033A61K35/12A01N63/12A61K35/62A61K38/17C07K14/435C12Q1/18
CPCA01N63/02C07K14/4354C12Q1/18A61K38/00A61K35/62A01N63/12
Inventor MANSFIELD, LINDA S.ABNER, SHEILA R.
Owner BOARD OF TRUSTEES OPERATING MICHIGAN STATE UNIV
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