Korelasi Gen Virulen gelE terhadap Pembentukan Biofilm dan Gelatinase pada Isolat Enterococcus faecalis

Abstract

Seratus dua puluh total sampel arsip digunakan pada penilitian ini. Sampel arsip terdiri atas 60 sampel asal inang ayam pedaging berupa sampel usap kloaka yang dikoleksi dari peternakan ayam pedaging di daerah Cianjur dan 60 sampel asal inang kucing berupa sampel usap anus (22 sampel) dan laring (38 sampel) dari pasien klinik hewan di Kota Depok dengan gejala infeksi saluran pencernaan. Penelitian ini bertujuan untuk mengetahui keberadaan faktor virulensi gelatinase yang dikodekan oleh gen virulen gelE serta menguji terkait korelasinya terhadap pembentukan biofilm dan aktivitas gelatinase pada isolat Enterococcus faecalis dari inang ayam dan kucing. Isolasi bakteri dilakukan dengan menggunakan media agar selektif diferensial KF Streptococcus. Konfirmasi molekuler menggunakan teknik polymerase chain reaction (PCR) dengan gen spesifik Efac. Deteksi faktor virulen gelatinase menggunakan gen virulen gelE. Uji fenotipik pembentukan biofilm menggunakan teknik spektrofotometri pada densitas optik 630 nm, sedangkan uji aktivitas gelatinase dilakukan dengan menginkubasi bakteri pada agar Brain Heart Infusion (BHI) dengan gelatin 4%. Analisis hasil dilakukan secara kuantitatif menggunakan uji chi-square dengan P < 0,05 dianggap signifikan secara statistik. Hasil isolasi bakteri didapatkan 21 dari 40 (52,50%) isolat dari usap kloaka ayam dan 10 dari 20 (50%) isolat dari usap anus kucing terkonfirmasi positif E. faecalis secara molekuler. Studi faktor virulensi menunjukkan bahwa 21 isolat (100%) ayam dan 10 isolat (100%) kucing E. faecalis terdeteksi memiliki gen virulen gelE. Isolat kloaka ayam dengan uji fenotipik biofilm didapatkan sebanyak 20 isolat (95,23%) dikategorikan positif kuat dan 1 isolat (4,76%) dikategorikan positif lemah, sedangkan pada isolat anus kucing didapatkan sebanyak 5 isolat (50%) dikategorikan positif kuat dan 5 isolat (50%) dikategorikan positif lemah. Uji fenotipik gelatinase pada isolat ayam adalah 17 isolat (80,95%) positif memiliki aktivitas gelatinase dan 4 isolat (19,04%) negatif gelatinase, sedangkan pada isolat kucing didapatkan sebanyak 6 isolat (60%) positif dan 4 isolat (40%) negatif aktivitas gelatinase. Kedua uji fenotipik memiliki nilai P < 0,05 atau memiliki hubungan yang signifikan antara keberadaan gen virulen gelE dengan pembentukan biofilm dan aktivitas gelatinase. Kesimpulannya adalah gen virulen gelE pada bakteri E. faecalis ditemukan pada semua isolat dari kloaka ayam dan usap anus kucing. Kedua jenis isolat tersebut diketahui mampu untuk pembentukan biofilm dan aktivitas gelatinase. Gen virulen gelE berkorelasi positif pada pembentukan biofilm lemah hingga kuat pada bakteri E. faecalis yang berasal dari isolat asal kloaka ayam dan anus kucing serta berkorelasi positif terhadap aktivitas gelatinase.

One hundred and twenty archival samples were used in this study. The archive samples consisted of 60 samples from broiler hosts in the form of cloacal swab samples collected from broiler farms in the Cianjur area and 60 samples from cat hosts in the form of anal swab samples (22 samples) and larynx swabs (38 samples) from veterinary clinic patients in Depok City with symptoms of gastrointestinal tract infections. This study aimed to explore the presence of gelatinase virulence factor encoded by the virulent gene of gelE and to examine its correlation to biofilm formation and gelatinase activity in Enterococcus faecalis isolates from chickens and cats. Bacteria isolation conducted by using KF Streptococcus selective-diffential agar media. Molecular confirmation using polymerase chain reaction (PCR) technique with the specific Efac gene. Detection of gelatinase virulent factor using the virulent gene of gelE. Phenotypic test of biofilm formation used spectrophotometric techniques at an optical density of 630 nm, while gelatinase activity was carried out by incubating bacteria on Brain Heart Infusion (BHI) agar with 4% gelatin. Result analysis was carried out quantitatively using the chi-square test with P < 0,05 considered statistically significant. The results of bacterial isolation were carried out 21 of the 40 (52,50%) isolates from chicken’s cloaca swab and 10 of the 20 (50%) isolates from cat’s anal swab molecularly confirmed positive for Enterococcus faecalis. The virulence factor study showed that 21 (100%) chicken’s isolates and 10 (100%) cat’s isolates of E. faecalis detected had the virulent gene of gelE. The isolates from chicken with biofilm formation test, 20 isolates (95,23%) strongly positive and 1 isolate (4,76%) weak positive, furthermore in cat’s anus isolates, 5 isolates (50%) strongly positive and 5 isolates (50%) weak positive. The gelatinase activity test on isolates from chicken were 17 isolates (80,95%) positive and 4 isolates (19,04%) negative, while isolates from cat were 6 isolates (60%) positive and 4 isolates (40%) negative for gelatinase activity. Both phenotypic tests had a P value < 0,05 or had a significant relationship between the presence of the gelE gene with biofilm formation and gelatinase activity. The conclusion was the gelE gene of E. faecalis bacteria was found in all isolates from chicken’s cloaca and cat’s anal swab. Both types of isolates were known to be able for biofilm formation and gelatinase activity. The virulent factor of gelE was positively correlated with weak to strong biofilm formation in Enterococcus faecalis bacteria from chicken’s cloaca and cat’s anal were positively correlated with gelatinase activity