Extractable surface proteins of indigenous probiotic strains confer anti-adhesion knack and protect against Methicillin-resistant Staphylococcus aureus induced epithelial hyperpermeability in HT-29 cell line

Abstract Lactic acid bacteria (LAB) were isolated from the bile, gizzard, intestine and caecum of the Malaysian domestic non-broiler chicken. Of the 56 isolates grown on MRS agar plates, 7 - namely, B3L3, B4L4, G5L5, B5L6, B10L7, I1L8, and C4L10 - showed inhibition against the methicillin resistant Staphylococcus aureus (MRSA). These isolates were all Gram positive cocci, non-spore forming, and catalase negative. The purified fraction from the three phase partitioning (TPP) method, employing t-butanol and ammonium sulfate, produced inhibitory zones of 8 to 18 mm in diameter on the indicator organism, Staphylococcus aureus (MRSA). The TPP purified fraction contained heat stable proteins of about 10 kDa in size and was inactivated by trypsin. Based on the partial rDNA sequences (>89% similarity), isolates B4L4 and G5L5 belonged to Enterococcus hirae; isolate B3L3 belonged to Enterococcus faecium; isolates B5L6, B10L7, I1L8 to Enterococcus faecalis; and isolate C4L10 to Enterococcus mundtii. These probiotic strains, exclusively isolated from the Malaysian non-broiler chicken, produced an antimicrobial protein or bacteriocin which was active against a pathogenic strain of methicillin resistant Staphylococcus aureus (MRSA).

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Rapid Detection of Epidemic Strains of Methicillin-Resistant Staphylococcus aureus

Journal of Clinical Microbiology ◽

10.1128/jcm.37.3.690-693.1999 ◽

1999 ◽

Vol 37 (3) ◽

pp. 690-693 ◽

Cited By ~ 3

Author(s):

Thomas A. Wichelhaus ◽

Sylvia Kern ◽

Volker Schäfer ◽

Volker Brade

Keyword(s):

Staphylococcus Aureus ◽

Rapid Detection ◽

Methicillin Resistant Staphylococcus Aureus ◽

Susceptibility Testing ◽

Protein A ◽

Surface Proteins ◽

University Hospital ◽

Chromosomal Dna ◽

Methicillin Resistant ◽

Specific Antibodies

Fifty methicillin-resistant Staphylococcus aureus(MRSA) initial isolates obtained from patients hospitalized in the orthopedic clinic of the Frankfurt University Hospital and 150 methicillin-sensitive Staphylococcus aureus (MSSA) isolates were investigated in this study to determine whether the Slidex Staph-Kit is capable of differentiating between MRSA and MSSA owing to its unique performance characteristics. The Slidex Staph-Kit is a combined latex hemagglutination test designed to detect clumping factor, protein A, and a specific surface immunogen for S. aureus. Clumping factor-positive strains cause erythrocytes sensitized with fibrinogen to hemagglutinate, thereby resulting in visible red clumps. S. aureus strains deficient in clumping factor agglutinate latex particles sensitized with specific antibodies against surface proteins of S. aureus, thereby resulting in visible white clumps. Our results demonstrate that white clumping has a 99% specificity as well as a 98% positive predictive value for MRSA. Clumping factor-negative MRSA, which have been reported to occur in several countries, are epidemic in the Frankfurt area and account for 80% of all MRSA initial isolates in the orthopedic clinic of the Frankfurt University Hospital. Genotyping of all MRSA isolates by macrorestriction analysis of chromosomal DNA revealed that 83% of clumping factor-negative MRSA are closely related to the “southern-German” epidemic strain. This is the first study demonstrating the Slidex Staph-Kit’s capability for identifying epidemic clumping factor-negative S. aureus strains as methicillin resistant even prior to antimicrobial susceptibility testing.

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