Identifying the bacteria causing ovine gangrenous mastitis and detection of Staphylococcus aureus in gangrenous milk by pcr

Bu çalışmanın amacı, koyun gangrenöz mastitisine neden olan bakterilerin identifiye edilmesi ve süt örneklerindeki Staphylococcus aureus’un polimeraz zincir reaksiyonu (PCR) ile tespit edilerek, koagülaz geni yönünden restriksiyon fragman uzunluğu polimorfizminin (RFLP) belirlenmesidir. Bu amaçla Şanlıurfa ilinde yetiştirilmekte olan gangrenöz mastitisli İvesi koyunlarından 264 adet süt örneği toplanmıştır. Bakteriyolojik incelemede Staphylococcus aureus (%75), Mannheimia haemolytica (%7.2), koagulaz negatif stafilakok (%6.43), Escherichia coli (%4.55), Corynebacterium spp. (%0.76) ve Actinomyces pyogenes (%0.76) identifiye edildi. Tesadüfi olarak seçilen 155 süt örneğinden bakteriyel DNA izole edildi. Koagulaz geni kullanılarak yapılan PCR amplifikasyonu ile örneklerin %91.6’sında S. aureus tespit edildi. Herbiri spesifik RFLP profili gösteren 420 (%0.7), 580 (%65.3) ve 780 (%34) baz çifti uzunluğunda 3 farklı PCR ürünü gözlendi. Sonuç olarak, S. aureus’un, bölgede yetiştirilen koyunlarda gangrenöz mastitislerden büyük oranda sorumlu olduğu bulunmuştur. S. aureus’un süt örneklerinden izole edilen DNA kullanılarak PCR metoduyla kolaylıkla tespit edilebileceği, iki coa geno-tipinin yaygın olduğu ve sınırlı sayıda suşun bölgedeki gangrenöz mastitis olgularının çoğundan sorumlu olduğu kanaatine varılmıştır.

Anahtar Kelimeler:

İvesi, bakteriyel hastalıklar, sığır meme yangısı, koagülaz negatif stafilokoklar, teşhis, koyun sütü, gıda bulaşıklılığı, gıda hijyeni, gıda güvenirliği, gen ifadesi, genler, genotip, meme yangısı, süt, süt hijyeni, doku ölümü, polimeraz zincir reaksiyonu, kısıtlı fragment uzunluğu polimorfizmi

Koyunlarda gangrenöz mastitise neden olan bakterilerin identifikayonu ve gangrenli sütlerdeki Staphylococcus aureus’un pcr ile tespit edilmesi

The objective of this study was to identify bacteria causing gangrenous ovine mastitis as well as to detect and genotype Staphylococcus aureus in milk samples by using polymerase chain reaction (PCR) amplification and restriction fragment length polymorphism (RFLP) of coagulase gene. A total of 264 milk samples were collected from Awassi sheep with gangrenous mastitis raised in Sanliurfa province of Turkey. By using bacteriological examination Staphylococcus aureus (75%), Mannheimia haemolytica (7.2%), coagulase negative staphylococci (6.43%), Escherichia coli (4.55%), Corynebacterium spp. (0.76%) and Actinomyces pyogenes (0.76%) were identified. Bacterial DNA was isolated from 155 randomly selected milk sample. By PCR amplification of coagulase gene S. aureus was detected in 91.6% of the milk samples. Three different amplicons of 420 (0.7%), 580 (65.3%) and 780 (34%) bp band sizes, each showing a specific RFLP profile were observed. In conclusion, S. aureus was found to be mostly responsible for gangrenous mastitis in sheep raised in this region. S. aureus in milk samples could be more rapidly detected by PCR method. Two coa genotype of S. aureus were found to be common suggesting that limited number of strains were responsible for most cases of gangrenous mastitis in the region under the study.

Keywords:

Awassi, bacterial diseases, bovine mastitis, coagulase negative staphylococci, diagnosis, ewe milk, food contamination, food hygiene, food safety, gene expression, genes, genotypes, mastitis, milk, milk hygiene, necrosis, polymerase chain reaction, restriction fragment length polymorphism,

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