Kajian faktor virulensi Staphylococcus aureus dari berbagai produk pangan asal ternakcNamiroh Setyo Anggraeni

ANGGRAENI, Namiroh Setyo, Prof. Dr. drh. Siti Isrina Oktavia Salasia

2008 | Tesis | S2 Sain Veteriner

Staphylococcus aureus mempunyai beberapa faktor virulensi yang potensial dalam menyebabkan penyakit pada manusia dan hewan. Enterotoksin yang dihasilkan oleh S. aureus apabila mencemari makanan dapat menyebabkan penyakit akibat keracunan makanan. Informasi mengenai berbagai faktor virulensi dan kemampuan S. aureus dalam memproduksi toksin, terutama keberadaan stafilokokal enterotoksin (SE) dalam produk pangan asal ternak belum banyak diungkap dan dilaporkan. Dalam penelitian ini dikaji berbagai faktor virulensi dan jenis toksin S. aureus yang diisolasi dari berbagai produk pangan asal ternak. Penelitian ini bertujuan untuk: (1) karakterisasi S. aureus, yang diisolasi dari berbagai produk pangan asal ternak, dan (2) deteksi enterotoksin dengan teknik polymerase chain reaction (PCR). Sampel yang digunakan adalah pangan asal ternak (susu segar dan daging ayam) yang diambil dari peternak dan pasar-pasar tradisional di daerah Yogya dan Solo, produk pangan asal ternak (susu kemasan, roti, keju, rolade, bakso dan sosis) yang diambil dari super market dan warung-warung tradisional. Karakterisasi S. aureus dilakukan berdasar produksi hemolisin, pigmen dan uji hemaglutinasi. Identifikasi molekuler dengan mengamplifikasi gen 23S rRNA menggunakan primer spesies spesifik. Deteksi gen enterotoksin dengan teknik PCR menggunakan primer spesifik dari berbagai jenis enterotoksin. Data yang diperoleh dari hasil karakterisasi, amplifikasi gen 23S rRNA dan gen enterotoksin dianalisis secara deskriptif dengan membandingkan karakter dan beberapa toksin S. aureus strain referen (kontrol positif) dan S. epidermidis yang tidak menghasilkan toksin (kontrol negatif). Berdasarkan hasil penelitian, sebanyak 32 isolat S. aureus dari susu segar, Yogya (5), susu segar, Solo (3), daging ayam, Yogya (5), susu kemasan, Yogya (4), roti, Yogya, Bandung, Jakarta (6), keju, Sidoarjo (1), rolade, Yogya (1), bakso, Yogya (1), sosis,Yogya (1) dan isolat asal manusia, Yogya (5) bersifat menfermentasi manitol, Gram positif, menghasilkan katalase dan koagulase positif. Hasil karakterisasi secara fenotipik diketahui bahwa S. aureus mampu memproduksi berbagai tipe hemolisin, yaitu tipe alfa (18,75%), beta (12,5%), gamma (62,5%) dan delta (6,2%). Staphylococcus aureus mampu memproduksi pigmen oranye (62,5%), kuning (21,87%) dan putih (15,62%), dan sebagian besar S. aureus mampu mengaglutinasi darah kelinci (96,87%). Berdasarkan identifikasi molekuler, semua isolat S. aureus dapat diamplifikasi terhadap gen 23S rRNA. Hasil evaluasi enterotoksin S. aureus terdapat lima isolat mengandung gen seb, lima isolat seh dan satu isolat mengandung kombinasi gen seb dan seh.

Staphylococcus aureus is a potential pathogen causing diseases in human and animals due to several virulence factors. Staphylococcal enterotoxins are responsible to foodborne disease by food contaminating toxins. The various virulence factors and the ability of S. aureus to produce toxins, especially existence of staphylococcal enterotoxins (SE) in various food of animal origins have not been reported. The research was done to study various virulence factors and enterotoxins produced by S. aureus isolated from various food of animal origins. The aims of the research were: (1) to characterize S. aureus isolated from various food of animal origins, and (2) to detect SE by using polymerase chain reaction (PCR) technique. The samples used in this study were collected from food of animal origins (fresh milks and chicken carcass) were obtain from the breeders and traditional markets in Yogya and Solo. Samples of processed food of animal origins (packaged milks, breads, cheeses, rolade, meat balls and sausages) were obtained from supermarkets and traditional markets. The characterizations of S. aureus were performed based on the production of hemolysins, pigments, and haemagglutination test. Molecular identification was conducted for the detection of the S. aureus 23S rRNA gene by using oligonecleotide species specific primers. The enterotoxins genes of S. aureus were investigated by using specific primers for enterotoxins sequences. The amplicons of 23S rRNA and enterotoxins gene of S. aureus were analyzed descriptively by comparing the virulence characteristics to S. aureus reference control (positive control) and S. epidermidis (negative control). The results of the studies showed that 32 isolates of S. aureus from fresh milks, Yogya (5), fresh milks, Solo (3), chicken carcass, Yogya (5), packaged milks, Yogya (4), breads, Yogya, Bandung, Jakarta (6), cheese, Sidoarjo (1), rolade, Yogya (1), meat ball, Yogya (1), sausage, Yogya (1) and isolates from human, Yogya (5) positive fermented mannitol, revealed Gram positive, catalase and coagulase positive. Phenotypically, S. aureus revealed an alfa (18,75%), beta (12,5%), gamma (62,5%) and delta (6,2%) -hemolysin on blood sheep agar, produced orange (62,5%), yellow (21,87%) and white (15,62%) pigments, and most of isolates could agglutinated erythrocytes of rabbit (96,87%). Staphylococcus aureus could be identified by using oligonucleotide species specific primers for 23S rRNA. Based on the detection of enterotoxins there were 5 isolates S. aureus containing genes seb, 5 isolates genes seh, and the combination of genes seb and seh was observed in 1 isolate.

Kata Kunci : Faktor virulensi,Staphylococcus aureus,Enterotoksin,Staphylococcus aureus, virulence factors, enterotoxins, food of animal origins