Deteksi enterotoksin dan karakterisasi staphylococcus aureus dari sampel susu sapi perah di daerah Kaliurang Yogyakarta

KHUSNAN, Prof. drh. Soesanto Mangkoewidjojo, M.Sc., Ph.D

2003 | Tesis | S2 Sain Veteriner

Staphylococcus aureus merupakan salah satu penyebab utama mastitis pada sapi perah yang menimbulkan kerugian ekonomi yang cukup besar akibat turunnya produksi susu. S. aureus dalam susu segar dan produk pangan menyebabkan toxic schock syndrome sebagai akibat dari keracunan pangan. Informasi mengenai kemampuan S. aureus dalam memproduksi toksin terutama enterotoksin di Indonesia belum banyak dilaporkan. Penelitian ini bertujuan untuk isolasi dan karakterisasi S. aureus secara fenotip dan deteksi adanya enterotoksin yang diproduksi S. aureus yang diisolasi dari susu sapi perah di peternakan sapi di Kaliurang Yogyakarta. Karakterisasi S. aureus meliputi uji clumping factor, uji koagulase, produksi hemolisin, produksi pigmen dan uji kepekaan S. aureus terhadap beberapa antibiotika. Deteksi enterotoksin dilakukan dengan teknik polymerase chain reaction (PCR) dengan menggunakan primer spesifik. Dari 22 sampel susu yang digunakan dalam penelitian ini berhasil diisolasi dan diidentifikasi 11 isolat S. aureus. Semua isolat positif pada uji clumping factor dan koagulase. Alfa-hemolisis dapat diamati pada 1 kultur, 2 kultur mempunyai sifat a dan b-hemolitik dan 9 kultur bersifat non-hemolitik. Berdasar produksi pigmen, 4 kultur menghasilkan pigmen berwarna oranye dan 7 kultur menghasilkan pigmen kuning. Hasil uji resistensi antibiotik terlihat bahwa 8 isolat (78,72%) resisten terhadap ampisilin atau penisilin dan hanya 3 isolat (27,27%) sensitif terhadap kedua antibiotika tersebut. Semua isolat S. aureus yang digunakan dalam penelitian ini diketahui sensitif (100%) terhadap klindamisin, minosiklin dan oksasilin dan bersifat intermedier (100%) terhadap gentamisin. Uji terhadap linkomisin tampak bahwa 8 S. aureus (72,72%) bersifat intermedier dan 3 isolat (27,27%) sensitif, terhadap tetrasiklin memperlihatkan sifat intermedier pada 10 isolat (90,91%) dan hanya 1 isolat (9,09%) bersifat sensitif. Diantara 11 kultur S. aureus yang digunakan dalam penelitian ini terdapat 3 kultur mengandung gen seg dan sej, 1 kultur mengandung gen seb dan seh. Keempat kultur S. aureus yang mengandung gen enterotoksin tersebut, mengandung pigmen berwarna oranye dan 3 diantaranya menghasilkan hemolisin. Karakteristik S. aureus isolat asal sapi perah di Kaliurang, Yogyakarta ini dapat digunakan sebagai penanda virulensi

Staphylococcus aureus is recognized as a major pathogen causing mastitis in dairy cows. The disease causes significant economic loss due to decreasing of milk production. Enterotoxin of S. aureus in the raw milk and food could cause a toxic shock syndrome and food borne poisoning. The ability S. aureus to produce toxins mainly enterotoxins in dairy cows in Indonesia have not been well reported. The research was designed to isolate and characterize S. aureus from milk samples of dairy cows in Kaliurang Yogyakarta and further detection of their enterotoxin. The characterization of S. aureus included clumping factor and coagulation assay, production of hemolysin and pigmen, and antibiotic sensitivity tests. The enterotoxins of S. aureus were detected by polymerase chain reaction (PCR) using specific primers. Eleven S. aureus isolates were identified from 22 milk samples. All 11 cultures were positive for clumping factors and coagulase. An a-hemolysis could be observed in 1 culture, a, b-hemolysis for 2 cultures and non-hemolysis in 9 cultures. Among 11 cultures revealed that 4 cultures produced an orange pigment and 7 cultures were yellow pigmented. The antibiotic resistance tests showed that 8 cultures (78,72%) were resistant to ampicilin or penicilin, only 3 cultures (27,27%) were susceptible to both antibiotics. All 11 cultures used in the investigation (100%) were susceptible to clindamycin, minocyclin dan oxacillin and intermediate to gentamicin. Eight cultures (72,72%) were intermediate and 3 cultures (27,27%) were susceptible to lincomycin, 10 cultures (90,91%) were intermediate and only 1 culture (9,09%) were susceptible to tetracycline. Among the 11 cultures used in this study 3 cultures harboured the gene seg and sei, and 1 culture the gene seb and seh. The 4 enterotoxin cultures were produced an orange pigments, three of them were produced hemolysin. This characteristics of S. aureus isolated from dairy cows in Kaliurang,Yogyakata could be used as a virulence marker.

Kata Kunci : Susu Sapi Perah,Enterotoksin dan Staphylococcus, Staphylococcus aureus, dairy cow milk, enterotoxin