UJI SENSITIVITAS DAN TIME KILL ASSAY BAKTERI ENTEROCOCCUS FAECALIS ATCC 29212 TERHADAP EKSTRAK ROYAL JELLY

RESTU AYU PUSPASARI, drg. Heni Susilowati, M.Kes., Ph.D ; Prof. Dr.drg. Juni Handajani, M.Kes., Ph.D

2022 | Skripsi | S1 HIGIENE GIGI

Bakteri Enteroccocus faecalis merupakan salah satu flora normal di dalam rongga mulut yang dapat menyebabkan infeksi saluran akar. Selain itu, faktor yang menyebabkan E. faecalis dapat bertahan hidup di dalam saluran akar sebagai organisme tunggal dan resisten terhadap obat-obat antimikrobial sehingga sulit dieliminasi dari saluran akar secara sempurna. Saat ini banyak dikembangkan penggunaan tanaman sebagai alternatif, dikarenakan adanya resistensi bakteri E. faecalis terhadap obat-obatan saluran akar terdahulu. Royal jelly mengandung senyawa-senyawa yang bersifat antibakteri, diantaranya major royal jelly protein dan royalisin. Tujuan penelitian ini adalah untuk mengetahui sensitivitas bakteri E. faecalis dan time kill assay ekstrak royal jelly terhadap E. faecalis ATCC 29212. Uji sensitivitas E. faecalis dilakukan dengan metode serial dilusi. Pada media BHI diinokulasikan suspensi E. faecalis dan dimasukkan ke microplate 96 sumuran. Larutan uji royal jelly dari setiap kelompok (n=4) dimasukkan ke dalam microplate sebanyak 100 µL. Setelah diinkubasi selama 24 jam pada suhu 37oC, sensitivitas dibaca menggunakan spektrofotometer untuk mendapatkan nilai densitas optik. Data diolah dengan uji One-way ANOVA dan Post-hoc Scheffe. Time-kill assay dilakukan dengan cara mencampurkan suspensi 100 µL E. faecalis dengan 1 ml larutan uji setiap kelompok (n=4). 100 µL cuplikan diambil setiap 24 jam dan 48 jam lalu ditanam pada media Mueller Hinton Agar. Setelah diinkubasi selama 24 jam pada suhu 37oC, dilakukan penghitungan jumlah koloni bakteri yang tumbuh pada agar. Data diolah dengan uji Kruskall-Wallis dilanjutkan Post-hoc Scheffe. Hasil uji sensitivitas menunjukkan kelompok ekstrak royal jelly dengan konsentrasi 25% menunjukkan adanya sensitivitas dan terdapat perbedaan signifikan (p<0,05). Hasil time-kill assay menunjukkan pada kelompok ciprofloxacin (kontrol positif) tidak terdapat adanya koloni E. faecalis yang tumbuh pada semua waktu. Hasil ini berbeda bermakna dengan kelompok ekstrak royal jelly 25% dan kontrol negatif. Ekstrak royal jelly terbukti bersifat antibakteri terhadap E. faecalis, waktu papar 48 jam ekstrak royal jelly 25% selama 48 jam lebih efektif menekan pertumbuhan bakteri daripada 24 jam.

Enterococcus faecalis is one of the normal flora in the oral cavity that can cause root canal infections. In addition, the factors that cause E. faecalis to survive in the root canal as a single organism and are resistant to antimicrobial drugs make it difficult to completely eliminate it from the root canal. Currently, the use of plants as an alternative has been widely developed, due to the resistance properties of E. faecalis and some of the disadvantages of previous root canal drugs. Royalisin compounds in royal jelly make royal jelly can be used as an inhibitor of bacterial growth. The purpose of this study was to determine the antibacterial effect of royal jelly extract and its effectiveness as a substitute for antibiotics in inhibiting the growth and time-kill assay of E. faecalis ATCC 29212. In this study, the sensitivity test of E. faecalis and time-kill assay was carried out. Sensitivity test was performed by serial dilution method. In BHI media, E. faecalis and put into microplate-wells. The test solution from each group (n=4) was added to microplate-wells 100 µL after being incubated for 24 hours at 37oC, the sensitivity was read using a spectrophotometer to see the results. Data were processed by One-way ANOVA and Post-hoc Scheffe. The time-kill assay was carried out by mixing 100 L of E. faecalis with 1 ml of the test solution for each group (n=4). 100 L of samples were taken every 24 hours and 48 hours and then planted on MHA media. After being incubated for 24 hours at 37oCcolonies E. faecalis counted.data were processed using the Kruskall-Wallis followed by Post-hoc Scheffe. The results of the sensitivity test showed that the royal jelly extract group with a concentration of 25% showed a clear zone or sensitivity and there was a significant difference (p<0.05) compared to the negative control group. The results of the time-kill assay showed that in the combination group there were no E. faecalis growing in all seconds. The results of the time-kill assay showed that in the positive control group (ciprofloxacin) there were no E. faecalis growing at all times. These results were significantly different from the 25% royal jelly extract group and the negative control group. Royal jelly 25% extract has antibacterial effect and is more effective at 48 hours exposure which can be used as dressing material in root canal treatment.

Kata Kunci : Royal jelly, Royalisin, Enterococcus faecalis, Uji sensitivitas, Time-kill assay.