This study aimed to detect methicillin resistant and slime producing Staphylococcus aureus in cases of bovine mastitis. A triplex PCR was optimized targetting 16S rRNA, nuc and mecA genes for detection of Staphylococcus species, S. aureus and methicillin resistance, respectively. Furthermore, for detection of slime producing strains, a PCR assay targetting icaA and icaD genes was performed. In this study, 59 strains were detected as S. aureus by both conventional tests and PCR, and 13 of them were found to be methicillin resistant and 4 (30.7%) were positive for mecA gene. Although 22 of 59 (37.2%) S. aureus isolates were slime-producing in Congo Red Agar, in PCR analysis only 15 were positive for both icaA and icaD genes. Sixteen and 38 out of 59 strains were positive for icaA and icaD gene, respectively. Only 2 of 59 strains were positive for both methicillin resistance and slime producing, phenotypically, suggesting lack of correlation between methicillin resistance and slime production in these isolates. In conclusion, the optimized triplex PCR in this study was useful for rapid and reliable detection of methicillin resistant S. aureus. Furthermore, only PCR targetting icaA and icaD may not sufficient to detect slime production and further studies targetting other ica genes should be conducted for accurate evaluation of slime production characters of S. aureus strains.

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SummaryIn this study, phenotypic and genotypic features of 10 L. garvieae strains isolated from rainbow trout farms were examined with 3 reference strains (Spain, England and ATCC 43921) comperativly. Rapid 32 STREP and conventional microbiologic tests were used for determining phenotipic features of L. garvieae strains. Altough there are diff erences, in Rapid 32 STREP system, between strains in terms of β-Glucuronidase, D-ribose, sorbitol, lactose, raffinose, alanyl-phenylalanyl-proline arylamidase, pyrrolidonyl arylamidase, hippurate hydrolysis, urease tests, all strains have been confirmed as L. garvieae by API web. In RAPD PCR analysis, in which ERIC 2 primer is used, L. garvieae isolates were genotyped within 3 separate clusters according to similarity coefficient index of 70%, and it was detected that a vast majority of isolates with Turkey-origin (8 isolates) belongs to predominant type LG1 genotype. In addition to this, antimicrobial tests of L. garvieae strains shows that there are resistance against gentamycin, neomycin, lincomycin, sulfamethoxazole-trimethoprim, oxytetracycline, erythromycin, amoxicillin, fl orfenicol and doxycycline, which are frequently used on fish in our country. Keywords: Oncorhynchus mykiss, Lactococcus garvieae, Rapid32 STREP, RAPD PCR, Antimicrobial sensitivity Lactococcus garvieae Suşlarının Fenotipik, Genotipik Karakterizasyonu ve Antimikrobiyal Duyarlılıklarının Belirlenmesi ÖzetAraştırmada, gökkuşağı alabalığı işletmelerinden izole edilmiş olan 10 adet Lactococcus garvieae suşunun 3 adet referans suşla (İspanya, İngiltere ve ATCC 43921) karşılaştırmalı olarak fenotipik ve genotipik özellikleri incelenmiştir. L. garvieae suşlarının fenotipik özelliklerinin belirlenmesinde konvansiyonel mikrobiyolojik ve Rapid 32 STREP testleri kullanılmıştır. Rapid 32 STREP sistemde suşlar arasında β-Glucuronidase, D-ribose, sorbitol, lactose, raffinose, alanyl-phenylalanyl-proline arylamidase, pyrrolidonyl arylamidase, hippurate hydrolysis, urease testleri yönüyle farklıklar olmasına rağmen API Web'de tüm suşlar L. garvieae olarak doğrulanmıştır. ERIC2 primerinin kullanıldığı RAPD PCR analizinde L. garveae izolatları %70 benzerlik katsayısına göre 3 farklı genotipe ayrılmış ve Türkiye kökenli izolatların büyük bir bölümü (8 izolat) predominant tip olan LG1 genotipine dahil olduğu belirlenmiştir. Ayrıca bu çalışmada L. garvieae suşlarının antibiyotik duyarlılık testlerinde ülkemizde balıklarda sıklıkla kullanılan gentamisin, neomycin, lincomycin, sulphamethoxazoletrimetoprim, oksitetrasiklin, eritromycin, amoxycillin, fl orfenikol ve doksisiklin'e karşı direnç geliştirmiş oldukları belirlenmiştir.

In this study, a total of 98 lactic acid bacteria isolated from rainbow trout intestines were screened for their probiotic properties. The isolates were tested for their ability to inhibit growth of Vagococcus salmoninarum and Lactococcus garvieae. Based on in vitro antagonism, 10 isolates were selected and evaluated pathogenicity in rainbow trout. Isolates were further investigated for hydrophobicity, bile salts and acid tolerance. These isolates were able to survive low pH and high bile concentrations and showed good adherence characteristics. Isolates were characterized phenotypically, and then, 16S rRNA gene sequence analysis was used for confirmation. Selected lactis M17 2-2, respectively. Our results suggest that these strains could provide an alternative for lactococcosis control in aquaculture. K E Y W O R D Saquaculture, lactic acid bacteria, Lactococcus garvieae, probiotic, rainbow trout, Vagococcus salmoninarum