Isolation of Staphylococcus simulans, a lysostaphin-producing organism, from bovine mastitis and its identification based on 16S rRNA sequencing method
Subject Areas : Applied Microbiology
Sajjad Yazdan Setad
1
(کارشناس ارشد، دانشگاه آزاد اسلامی، واحد دامغان، دانشکده علوم زیستی، گروه میکروبیولوژی)
Majid Moghbeli
2
(استادیار، دانشگاه آزاد اسلامی، واحد دامغان، دانشکده علوم زیستی، گروه میکروبیولوژی)
Hatef Ajoudanifar
3
(استادیار، دانشگاه آزاد اسلامی، واحد دامغان، دانشکده علوم زیستی، گروه میکروبیولوژی)
Keywords: 16S rRNA, Staphylococcus simulans, Lysostaphin, Bovine mastitis,
Abstract :
Background and Objectives: S. simulans is a gram-positive cocci that can be isolated from skin and urine, as well as some clinical samples in human and animals, such as bovine mastitis. The present study was aimed to isolate and identify S. simulans, a lysostaphin-producing organism, from bovine mastitis and to test therapeutic property of this species against the staphylococcal infections. Materials and Methods: This cross- sectional study was done on the collected samples from breast of the cows afflicted to mastitis in Damghan, Semnan Province. The samples were cultured on nutrient agar and were examined based on Gram staining and biochemical tests. Then, the genomic DNA of the samples suspected for S. simulans were extracted. The accurate identification of the bacteria and tracking of lysostaphin gene were evaluated by using PCR and sequencing. Results: One out of 61 gram-positive isolated cocci from bovine mastitis samples was detected as S. simulans. This result was verified using 16S rRNA sequencing. Conclusion: This study is the first report of isolation of S. simulans, a lysostsphin-producing organism, from bovine mastitis. This species is potentially useful for extraction of lysostsphin which is applied for treatment of the infections caused by antibiotic resistance bacteria.
1. Bergey D, Breed R, Murray E, Ilitchens A. Bergey's Manual of Determinative Bacteriology. 5th ed. Baltimore. Williams and Wilkins. 1939. P: 142-189.
2. Schindler C, Schuhardt V. Purification and properties of lysostaphin: a lytic agent for Staphylococcus aureus. Biochem Biophys Acta. 1965; 97: 242-250.
3. Schindler C, Schuhardt V. Lysostaphin: a new bacteriolytic agent for the Staphylococcus. Proc Natl Acad Sci USA. 1964; 51: 414-421.
4. DeHart H, Heath H, Heath L, LeBlanc P, Sloan G. The lysostaphin endopeptidase resistance gene (epr) specifies modification of peptidoglycan cross bridges in Staphylococcus simulans and Staphylococcus aureus. Appl Environ Microbiol. 1995; 61: 1475-1479.
5. Adegoke A, Komolafe A. Multi-drug resistant Staphylococcusaureus in clinical cases in Ile-Ife, Southwest Nigeria. Int J Med Sci. 2009; 1(3): 68-72.
6. Parsonnet J, Deresiewecz R, Braunwald E, Fauci AS. Staphylococcal infections. 15th ed. Harrison's Principles of Internal Medicine. 2001. P: 889- 901.
7. Bastos M, Coutinho BG, Coelho M. Lysostaphin: a Staphylococcal bacteriolysin with potential clinical applications. Pharmaceutic. 2010; 3(4): 1139-1161.
8. Taponen S, Simojoki H, Haveri M, Larsen H, Pyorala S. Clinical characteristics and persistence of bovine mastitis caused by different species of coagulase-negative staphylococci identified with API or AFLP. Vet Microbiol. 2006; 115: 199-207.
9. Kado C, Liu S. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981; 145: 1365-1373.
10. Sambrook J, Russell D. Molecular cloning: a laboratory manual. 3rd ed. Cold Spring Harbor Laboratory Press. 2001. P: 245-299.
11. Kloos W. Natural populations of the genus Staphylococcus. Ann Rev Microbiol. 1980; 34: 559-592.
12. Kloos W, Schleifer K. Isolation and characterization of staphylococci from human skin II. Descriptions of four new species: Staphylococcus warneri, Staphylococcus capitis, Staphylococcus hominis and Staphylococcus simulans. Int J Syst Bacteriol. 1975; 25: 62-79.
13. Angeliki R. Burriel. Isolation of coagulase-negative staphylococci from the milk and environment of sheep. J Dairy Res. 1998; 65: 139-142.
14. Heng N, Wescombe P, Burton J, Jack R, Tagg J. The diversity of bacteriocins in gram-positive bacteria. Springer: New York, NY, USA. 2007; P: 45-92.
15. Sloan G, Robinson J, Kloos W. Identification of ‘Staphylococcus staphylolyticus’ NRRL B-2628 as a biovar of Staphylococcus simulans. lnf J Syst Bacteriol. 1982; 32: 170-174.
16. Aarestrup F, Larsen H, Jensen N. Characterization of Staphylococcus simulans strains isolated from cases of bovine mastitis. Vet Microbiol. 1999; 66: 165-170.
17. Casaburi A, Blaiotta G, Mauriello G, Pepe O, Villani F. Technological activities of Staphylococcus carnosus and Staphylococcus simulans strains isolated from fermented sausages. Meat Sci. 2005; 71: 643-650.
18. Pyorala S, Taponen S. Coagulase-negative staphylococci: emerging mastitis pathogens. Vet Microbial. 2008; 134(1-2): 3-8.