Seyhan KAYNARCA, SÜHEYLA TÜRKYILMAZ

Sığır mastitislerinden izole edilen stafilokoklarda metisilin direnci ve slaym pozitifliği

Bu çalışmanın amaçları sığır mastitislerine neden olan stafilokok suşlarında metisilin direnci ve slaym oluşumunun incelenmesidir. Materyal olarak, 152 sağmal inekten alınan 339 mastitisli süt örneği kullanılmıştır. İzole edilen mikroorganizmaların cins düzeyinde identifikasyonu standart biyokimyasal yöntemlerle; koagulaz pozitif stafilokok (KPS) ve koagulaz negatif stafilokok (KNS) suşlarının tür düzeyinde identifikasyonları ise sekans analizi ile yapılmıştır. Stafilokok suşlarının metisilin dirençleri sefoksitin diski kullanılarak disk diffüzyon yöntemiyle, slaym varlığı ise Kongo Red Agar yöntemine göre belirlenmiştir. Mastitis etkeni olarak ilk sırayı KNS’lar (%24.5) alırken, bunu KPS’lar (%20.9) takip etmiştir. Bunlardan 16 (%10.4) suş metisilin dirençli, 55 (%35.7) suş slaym pozitif olarak saptanırken; KPS ve KNS suşlarında slaym oluşumunun metisilin direncinde artışa yol açmadığı tespit edilmiştir. Metisilin direncinin Aydın yöresinde bir sorun oluşturmaya başladığı, bundan sonraki yıllarda stafilokokal mastitis tedavisinde metisilin direnci ve slaym üretiminin dikkate alınması gerekeceği düşünülmektedir.

Anahtar Kelimeler:

süt ineği, biyokimya, biyofilm, sığır meme yangısı, koagülaz negatif stafilokoklar, süt sığırı, ilaç direnci, meme yangısı, metisilin, süt, nükleotit dizileri

Methicillin resistance and slime positivity of staphylococci isolated from bovine mastitis

The aims of this study were to determine the slime formation and methicilline resistance in isolated staphylococcus species from bovine mastitis. Material of this study was consisted of 339 milk samples with mastitis taken from 152 dairy cattle rearing. Identifications of isolated microorganisms on genus basis were performed with standart biochemical methods while sequence analysis was used for species basis identification of coagulase negative (CNS) and coagulase positive (CPS) staphylococcus strains. Methicilline resistance of stapylococcus strains were performed with disk diffusion method by using cefoxitin disk and slime formation was detected with Congo Red Agar method. CNS were the most common cause (24.5%) of mastitis in cows while CPS (S. aureus) were the most second (20.9%). 16 (10.4%) staphylococcus strains were found as methicilline resistance while 55 (35.7%) were slime positive from 154 stapylococcus strains. It was determined that slime formation has no effect on methicilline resistance in CNS and CPS staphylococcus strains. It was thought that methicilline resistance is beginning a problem, methicilline resistance and slime production may be important factors in the treatment of staphylococcal mastitis in the near future in Aydın.

Keywords:

dairy cows, biochemistry, biofilms, bovine mastitis, coagulase negative staphylococci, dairy cattle, drug resistance, mastitis, methicillin, milk, nucleotide sequences,

PDF

___

1. Hartman BJ, Tomas A: Low-affinity penicillin-binding protein associated with-lactam resistance in Staphylococcus aureus. J Bacteriol, 158, 513-516, 1984.
2.Chambers HF: Methicillin resistance in staphylococci: Molecular and biochemical basis and clinical implications. Clin Microbiol Rev, 10, 781-791, 1997.
3.Leonard FC, Markey BK: Meticillin-resistant Staphylococcus aureus in animals: A Review. Vet J, 175, 27-36, 2008.
4.Morgan M: Methicillin-resistant Staphylococcus aureus and animals: zoonosis or humanosis. J Antimicrob Chemoth, doi: 10.1093/jac/dkn405, 2008.
5.Chai H, Archambault M, Prescott JF: 16S ribosomal RNA sequence-based identification of veterinary clinical bacteria. J Vet Diagn Invest, 15, 465-469, 2003.
6.Telli M, Sümerkan B, Eşel D: Staphylococcus aureus’ta metisilin direncinin belirlenmesinde sefoksitin disk, oksasilin disk, oksasilin agar tarama ve PB2a lateks testlerinin karşdlaştdrdlmasd. İnfek Derg, 20, 93-96, 2006.
7.Cengiz AS, Us E, Cengiz AT: The clinical importance of slime production. Inönü Univ Tdp Fak Derg, 13, 193-197, 2006.
8.Arciola CRA, Campoccia D, Baldassarri L, Donati ME, Pirini V, Gamberini S, Montanaro L: Detection of biofilm formation in Staphylococcus epidermidis from implant infections. Comparison of a PCR-method that recognizes the presence of ica genes with two classic phenotypic methods. J Biomed Mater Res A, 76, 425-430, 2006.
9.Grinholc M, Wegrzyn G, Kurlenda J: Evaluation of biofilm production and prevalance of the icaD gene in methicillinresistant and methicillin-susceptible Staphylococcus aureus strains isolated from patients with nasocomial infections and carriers. FEMS Immunol Med Microbiol, 50, 375-379, 2007.
10.Juhász-Kaszanyitzky É, Jánosi S, Somogyi P, Dán Á, Bloois L G, Duijkeren E, Wagenaar JA: MRSA transmission between cows and humans. Emerg Infect Dis, 13, 630-632, 2007.
11.Rich M, Roberts L, Kearns AM: Methicillin-resistant Staphylococcus aureus isolates from companion animals. Vet Microbiol, 105, 313-314, 2005.
12.Türkyılmaz S, Tekblylk S, Oryasin E, Bozdoğan B: Molecular epidemiology and antimicrobial resistance mechanisms of methicillin-resistant Staphylococcus aureus isolated from bovine milk. Zoonoses Public Health, 57, 197-203, 2010.
13.Ak S, Horoz H, Ilgaz A: Trakya bölgesinde sığır mastitislerinden sorumlu bulaşdcd ve çevresel bakteriyel etkenler ve antibiyotik duyarlılıkları. Istanbul Univ Vet Fak Derg, 26, 353365, 2000.
14.Güler L, Ok Ü, Gündüz K, Gülcü Y, Hadimli HH: Antimicrobial susceptibility and coagulase gene typing of Staphylococcus aureus isolated from bovine clinical mastitis cases in Turkey. J Dairy Sci, 88, 3149-3154, 2005.
15.Kırkan Ş, Göksoy EÖ, Kaya O: Identification and antimicrobial susceptibility of Staphylococcus aureus and coagulase negative staphylococci from bovine mastitis in the Ayddn region of Turkey. Turk J Vet Anim Sci, 29, 791-796, 2005.
16.Kireçci E, Çolak A: Kuru dönem başlangdcdnda subklinik mastitisli ineklerden izole edilen stafilokok suşlardnda metisilin direnci. Kafkas Univ Vet Fak Derg, 8, 98-100, 2002.
17.Baştan A: İneklerde Meme Hastaldklard. s. 25-78. Hatipoğlu Basdm ve Yaydm San. Tic. Ltd. Şti., Ankara, 2002.
18.Koneman EW, Allen SD, Janda WM, Schreckenberger PC, Winn WC: Color Atlas and Textbook of Diagnostic Microbiology. The Gram-positive cocci: Part-1: Staphylococci and related organisms. pp. 539-576. Lippincott, New York, USA, 1997.
19.Sghir A, Antonopoulos D, Mackie RI: Design and evaluation of a lactobacillus group-specific ribosomal RNA-targeted hybridization probe and its application to the study of intestinal microecology in pigs. Syst Appl Microbiol, 21, 291296, 1998.
20.Suau A, Bonnet R, Sutren M, Godon JJ, Gibson G, Collins MD, Dore´J: Direct rDNA community analysis reveals a myriad of novel bacterial lineages within the human gut. Appl Environ Microbiol, 65, 4799-4807, 1999.
21.Freeman, DJ, Falkiner FR, Keane CT: New method for detecting slime producing by coagulase negative staphylococci. J Clin Pathol, 42, 872-874, 1989.
22.Bauer AW, Kirby WM, Sherris JC, Turck M: Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol, 45, 493, 1966.
23.Anonim: CASFM: Comite de l'antibiogramme de la Societe francaise de microbiologie: Communique, http://www.sfm.asso.fr/nouv/general.php?pa=2, Accessed: 16.06.2009.
24.Anonim: CLSI: Clinical and Laboratory Standards Institute Performance Standards for Antimicrobial Susceptibility Testing. 15th informational supplement. Approved Standard MS100-S16. Wayne, PA, USA, 2006.
25.Sümbüloğlu V, Sümbüloğlu K: Biyoistatistik. s. 156-174. Özdemir Yaydncdldk, Ankara, 1993.
26.Bradley AJ: Bovine mastitis: An evolging disease. Vet J, 164, 116-128, 2002.
27.Bayar S: Süt örneklerinden staphylococcus ve streptococcus türlerinin izolasyonu, identifikasyonu ve antibiyotik dirençlerinin belirlenmesi. Yüksek Lisans Tezi. Kahramanmaraş Sütçü İmam Üniv Fen Bil Enst Biyoloji AD, Kahramanmaraş, 2007.
28.Gülcü HB, Ertaş HB: Bacteriological investigation of udder lobes of cows with mastitis slaughtered in the Elazdğ region. Tr J Vet Anim Sci, 28, 91-94, 2002.
29.Tel OY, Keskin O, Zonturlu KA, Kaya NBA: Şanldurfa yöresinde subklinik mastitislerin görülme orand, aerobik bakteri izolasyonu ve duyarld antibiyotiklerin belirlenmesi. Fdrat Üniv Sağ Bil Vet Derg, 23, 101-106, 2009.
30.Taponen S, Pyorala S: Coagulase-negative staphylococci as cause of bovine mastitis-not so different from Staphylococcus aureus? Vet Microbiol, 134, 29-36, 2009.
31.Elçi S, Gül K, Özel F, Suay A, Mete Ö: Koagülaz negatif stafilokoklarda makro ve mikro yöntemle “slaym” oluşumunun saptanmasd ve antibiyotik direncinin araştdrdlmasd. İnfek Derg, 10, 203-206, 1996.
32.Çelik İ, Cihangiroğlu M, Çabalak M, Sevim E, Akbulut A, Klllş SS: Hastane kaynaklı koagulaz negatif stafilokoklarda fosfamisin duyarlılığı ile metisilin direnci ve slaym yapdmd arasdnda ilişki. Ankem Derg, 19, 139-143, 2005.
33.Çelik İ, Cihangiroğlu M, Sevim E, Çabalak M, Akbulut A: Sağlık çalışanlarının burunlarından izole edilen koagulaz pozitif ve koagulaz negatif stafilokoklarda metisilin direnci ve slaym pozitifliği. Fdrat Tdp Derg, 10, 123-126, 2005.
34. Fındık A, Akan N, Onuk EE, Çakıroğlu D, Çiftçi A: Methicillin resistance profile and molecular typing of Staphylococcus aureus strains isolated from noses of the healthy dogs. Kafkas Univ Vet Fak Derg, 15, 925-930, 2009.