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    PCR and ELISA for staphylococcal enterotoxins and detection of some exotoxins from Staphylococcus spp. strains by PCR
    (Polish Soc Veterinary Sciences Editorial Office, 2016) Kahya, Serpil; Guran, Husnu Sahan; Yilmaz, Ozge
    The aims of this study were to determine the existence of some staphylococcal enterotoxin (SE) (sea, seb, sec, sed, and see) proteins and genes in coagulase-positive staphylococci (CPS) by ELISA and PCR and to assess SE-like toxin (SEI) (seg, seh, sei, sej, sem, sen, and seo), exfoliative toxin (eta and etb), toxic shock syndrome toxin-1 (tst) and 16S rRNA genes in 11 different Staphylococcus strains [90 CPS and 118 coagulase-negative staphylococci (CNS)] isolated from 250 ground meat samples by either monoplex or multiplex PCR. SEs were identified in 36 (40%) out of 90 CPS isolates by both ELISA and PCR, with the following distribution: sea was identified in 7 (7.7%), seb in 5 (5.5%), sec in 3 (3.3%), sed in 4 (4.4%), and see in 17 (18.8%). In addition, a total of 90 CPS and 118 CNS isolates were investigated for the presence of 11 SE, SEI, eta-etb, tst, and 16S rRNA genes. Overall, 145 (69.7%) of the Staphylococcus spp. isolates tested positive for one or more toxin genes. These results indicate that CNS may play an important role in food poisoning and that SEI toxins must be investigated in greater detail in future studies of both CPS and CNS.
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    Species Diversity and Pheno- and Genotypic Antibiotic Resistance Patterns of Staphylococci Isolated from Retail Ground Meats
    (Wiley, 2015) Guran, Husnu Sahan; Kahya, Serpil
    The presence and species diversity of staphylococci in 250 ground beef and lamb meat samples obtained from Diyarbakir, Turkey were investigated. The presence of the 16S rRNA gene, mecA, nuc, pvl, and femA was analyzed by multiplex PCR. Pheno- and genotypic antibiotic resistance profiles of 208 staphylococci isolates were established. Of the ground beef and ground lamb samples, 86.4% and 62.4% were positive for staphylococci, respectively. Staphylococcus aureus, S. saprophyticus, S. hominis, S. lentus, S. pasteuri, S. warneri, S. intermedius, and S. vitulinus made up 40.8%, 28.8%, 11%, 3.8%, 3.8%, 2.4%, 2.4%, and 2.4% of isolates, respectively. Of the 85 S. aureus isolates, 40%, 47%, and 5.8% carried femA, mecA, and pvl, respectively, whereas the corresponding rates for the 118 coagulase-negative staphylococci (CoNS) were 0%, 10.1%, and 0%, respectively. We determined from the 208 isolates, the highest antibiotic resistances were to tetracycline and oxytetracycline (85.5%), followed by penicillin (51.4%), novobiocin (45.6%), ampicillin (39.9%), and doxycycline (31.7%), using the Clinical and Laboratory Standards Inst. (CLSI) method. All isolates were sensitive to gentamycin, ofloxacin, and tobramycin, but 2.3% of the S. aureus isolates had resistance to vancomycin. The staphylococci isolates carried tet(K), blaZ, tet(L), tet(W), cat, tet(S), tet(M), ermB, ermA, and ermC antibiotic resistance genes at rates of 59%, 51.7%, 36.9%, 31.8%, 27.2%, 27.2%, 24.4%, 18.1%, 7.9%, and 3.9%, respectively. Practical Application Our study is the 1st to detect pvl in MRSA isolates from ground meat samples in Turkey. Moreover, we detected 2 instances of vancomycin resistance in MRSA isolates and also detected a high level of multidrug resistance in staphylococci isolates. The results of this study show that ground meat can be a source of staphylococci of concern to public health because of potential for pathogenicity and as a reservoir for antibiotic resistance genes.