Synergistic hemolysins of coagulase-negative staphylococci (CoNS)
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A total of 104 coagulase negative staphylococci, belonging to S. capitis, S. hominis, S. haemolyticus and S. warneri, originating from the collection of the Department of Pharmaceutical Microbiology (ZMF), Medical University of Lodz, Poland, were tested for their synergistic hemolytic activity. 83% of strains produced δ-hemolysin, however, the percentage of positive strains of S. haemolyticus, S. warneri, S. capitis and S. hominis was different - 98%, 78%, 75% and 68%, respectively. Highly pure hemolysins were obtained from culture supernatants by protein precipitation with ammonium sulphate (0-70% of saturation) and extraction by using a mixture of organic solvents. The purity and molecular mass of hemolysins was determined by TRIS/Tricine PAGE. All CoNS hemolysins were small peptides with a molar mass of about 3.5 kDa; they possessed cytotoxic activity against the line of human foreskin fibroblasts ATCC Hs27 and lysed red cells from different mammalian species, however, the highest activity was observed when guinea pig, dog and human red blood cells were used. The cytotoxic effect on fibroblasts occurred within 30 minutes. The S. cohnii ssp. urealyticus strain was used as a control. The antimicrobial activity was examined using hemolysins of S. capitis, S. hominis, S. cohnii ssp. cohnii and S. cohnii ssp. urealyticus. Hemolysins of the two S. cohnii subspecies did not demonstrate antimicrobial activity. Cytolysins of S. capitis and S. hominis had a very narrow spectrum of action; out of 37 examined strains, the growth of only Micrococcus luteus, Corynebacterium diphtheriae and Pasteurella multocida was inhibited.
- Allaker RP, Whitlock M, Lloyd DH (1991) Cytotoxic activity of Staphylococcus hyicus. Vet Microbiol 26: 161-166.
- Bhakdi S (1985) Positive camp-phenomenon elicited by coagulase-negative staphylococci. Med Microbiol Immunol 174: 275-279.
- Bohach GA, Dinges MM, Mitchel DT, Ohlendorf DH, Schlievert PM (1997) Exotoxins. In The Staphylococci in Human Disease. Crossley KB, Archer GL, eds, pp 83-111. Churchil Livingstone, New York.
- Beaudet RJ, Bisaillon S, Saheb A, Sylvestre M (1982) Production, purification, and preliminary characterization of a gonococcal growth inhibitor produced by a coagulase-negative staphylococci isolated from a urogenital flora. Antimicrob Agents Chemother 22: 277-283.
- Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.
- Butt HL, Dunstan RH, McGregor NR, Roberts TK, Zerbes M, Klineberg IJ (1998) An association of membrane-damaging toxins from coagulase-negative staphylococci and chronic orofacial muscle pain. J Med Microbiol 47: 577-584.
- Cheung GY, Joo HJ, Chatterjee SS, Otto M (2014) Phenol-soluble modulins - critical determinants of staphylococcal virulence. FEMS Microbiol Rev 38: 698-719.
- Dinges MM, Orwin PM, Schlievert PM (2000) Exotoxins of Staphylococcus aureus. Clin Microbiol Rev 13: 16-34.
- Donvito B, Etienne J, Denoroy L, Greenland T, Benito Y, Vandenesch F (1997) Synergistic hemolytic activity of Staphylococcus lugdunensis is mediated by three peptides encoded by non-agr genetic locus. Infect Immun 65: 95-100.
- Fitton JE, Dell A, Shaw WV (1980) The amino acid sequence of the delta haemolysin of Staphylococcus aureusi. FEBS Letters 115: 209-212.
- Foster TJ (2002) Staphylococcus aureus. In Molecular Medical Microbiology, Sussman M, eds, pp 839-888. Academic Press, London, San Diego.
- Frenette M, Beaudet R, Bisaillon JG, Sylvestre M, Portelance V (1984) Chemical and biological characterization of on a gonococcal growth inhibitor produced by Staphylococcus haemolyticus isolated from urogenital flora. Infect Immun 46: 340-345.
- Heatley NG (1971) A new method for the preparation and some properties of staphylococcal delta-haemolysin. J Gen Microbiol 69: 269-278.
- Hebert GA, Hancock GA (1985) Synergistic hemolysis exhibited by species of staphylococci. J Clin Microbiol 22: 409-415.
- Hebert GA, Cooksey RC, Clark NC, Hill BC, Jarvis WR, Thornsberry C (1988a) Biotyping coagulase-negative staphylococci. J Clin Microbiol 26: 1950-1956.
- Hebert GA, Crowder CG, Hancock GA, Jarvis WR, Thornsberry C (1988b) Characteristics of coagulase-negative staphylococci that help differentiate these species and other members of the family Micrococcaceae. J Clin Microbiol 26: 1939-1949.
- Hebert GA (1990) Hemolysins and other characteristics that help differentiate and biotype Staphylococcus lugdunensis and Staphylococcus schleiferii. J Clin Microbiol 28: 2425-2431.
- Hechard Y, Ferraz S, Bruneteau E, Steinert M, Berjeaud JM (2005) Isolation and characterization of a Staphylococcus warneri strain producing an anti-Legionella peptide. FEMS Microbiol Lett 252: 19-23.
- Heikens E, Fleer A, Paauw A, Florijn A, Fluit AC (2005) Comparison of genotypic and phenotypic methods for species-level identification of clinical isolates of coagulase-negative staphylococci. J Clin Microbiol 43: 2286-2290.
- Hertle R, Hilger M, Weingardt-Kocher S, Walev I (1999) Cytotoxic action of Serratia marcescens hemolysin on human epithelial cells. Infect Immun 67: 817-825.
- Kreger AS, Kim K, Zaboretzky F, Bernheimer AW (1971) Purification and properties of staphylococcal delta-hemolysin. Infect Immun 3: 449-465.
- Mak P, Maszewska A, Różalska M (2008) The amino acid sequences and activities of synergistic hemolysins from Staphylococcus cohnii. FEMS Microbiol Lett 287: 230-235.
- Marchand A, Verdon J, Lacombe C, Crapart S, Hechard Y, Berjeaud JM (2011) Anti-Legionella activity of staphylococcal hemolytic peptides. Peptides 32: 845-851.
- Marchand A, Augenstreich J, Loiseau C, Verdon J, Lecomte S, Berjeaud JM (2015) Effect of amino acid substitution in the staphylococcal peptides warnericin RK and PSMα on their anti-Legionella and hemolytic activities. Mol Cell Biochem 405: 159-167.
- McKevitt AI, Bjornson GL, Mauracher CA, Scheifele DW (1990) Amino acid sequence of delta-like toxin from Staphylococcus epidermidis. Infect Immun 58: 1473-1475.
- Molnar C, Hevessy Z, Rozgonyi F, Gemmell CG (1994) Pathogenicity and virulence of coagulase negative staphylococci in relation to adherence, hydrophobicity and toxin production. J Clin Pathol 47: 743-748.
- Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63.
- Otto M (2004) Virulence factors of the coagulase-negative staphylococci. Front Biosci 9: 841-863.
- Różalska M, Józefowicz-Korczyńska M (2001) Microflora of the posterior pharyngeal wall, larynx and postoperative wounds in patients after laryngectomy. Med Dośw Mikrobiol 53: 63-70 (in Polish).
- Różalska M, Szewczyk E, Piotrowski A (1993) Bacterial flora of the hospital environment: identification, susceptibility to disinfecting agents, antibiotics and chemotherapeutics. Med Dośw Mikrobiol 45: 373-378 (in Polish).
- Różalska M, Szewczyk EM (2008) Staphylococcus cohnii hemolysins - isolation, purification and properties. Folia Microbiol (Praha) 53: 521-526.
- Schagger H, von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166: 368-379.
- Scheifele DW, Bjornson GL, Dyer RA, Dimmick JE (1987) Delta-like toxin produced by coagulase-negative staphylococci is associated with neonatal necrotizing enterocolitis. Infect Immun 55: 2268-2273.
- Scheifele DW, Bjornson GL (1988) Delta toxin activity in coagulase-negative staphylococci from the bowels of neonates. J Clin Microbiol 26: 279-282.
- Szewczyk EM, Piotrowski A, Różalska M (2000) Predominant staphylococci in the intensive care unit of a paediatric hospital. J Hosp Infect 45: 145-154.
- Verdon J, Berjeaud JM, Lacombe C, Hechard Y (2008) Characterization of anti-Legionella activity of warnericin RK and delta-lysin I from Staphylococcus warneri. Peptides 29: 978-984.
- Verdon J, Falge M, Maier E, Bruhn H, Steinert M, Faber C, Benz R, Hechard Y (2009a) Detergent-like activity and α-helical structure of warnericin RK, an anti-Legionella peptide. Biophys J 97: 1933-1940.
- Verdon J, Girardin N, Lacombe C, Berjeaud JM, Hechard Y (2009b) δ-hemolysin, an update on a membrane-interacting peptide. Peptides 30: 817-823.
- Watson DC, Yaguchi M, Bisaillon JG, Beaudet R, Morosoli R (1988) The amino acid sequence of a gonococcal growth inhibitor from Staphylococcus haemolyticus. Biochem J 252: 87-93.
- Watts JL, Owens WE (1987) Synergistic hemolysis associated with coagulase negative staphylococci isolated from bovine mammary glands. J Clin Microbiol 25: 2037-2039.
- Wiseman GM (1975) The hemolysins of Staphylococcus aureus. Bacteriol Rev 39: 317-344.
- Wladyka B, Wielebska K, Wloka M, Bochenska O, Dubin G, Dubin A, Mak P (2013) Isolation, biochemical characterization, and cloning of a bacteriocin from the poultry-associated Staphylococcus aureus strain CH-91 Appl Microbiol Biotechnol 97: 7229-7239.
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