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UJI AKTIVITAS ANTIBAKTERI HASIL HIDROLISIS ENZIMATIS MINYAK KELAPA MURNI TERHADAP SALMONELLA TYPHI DAN
SALMONELLA TYPHIMURIUM SECARA IN-VITRO DAN IN-VIVO
Abstrak
Salmonellosis merupakan salah satu penyakit zoonosis yang disebabkan oleh bakteri patogen Salmonella melalui kontaminasi silang makanan (foodborne disease). Pengobatan Salmonellosis menggunakan antibiotik selain mengoreksi dehidrasi dan gangguan elektrolit.Minyak kelapa murni (Virgin Coconut Oil, VCO) mempunyai potensi medis seperti sifat antibakteri, antivirus, dan antijamur dari kandungan asam lemak rantai sedang (Medium Chain Fatty Acid, MCFA), khususnya asam laurat (C12:0) atau dalam bentuk monogliserida (monolaurin atau ML) yang terdapat didalamnya. Tujuan penelitian ini adalah untukmenguji aktivitas antibakteri hasil hidrolisis enzimatis VCO (VCOH) yang menghasilkan kombinasiasam laurat dan monolaurin terhadap Salmonella secara vitro dan in-vivo yang diharapkan memberikan efek sinergis.
Sampel VCO yang digunakan adalahPalem Mustika VCO-Virgin Coconut Oilyang diproduksi oleh Siti Nurbaya, Sumatera Barat. VCO dihidrolisis dengan enzim lipase (LIPOZYME® TL IM) selama 18 jam dengan interval waktu pengamatan nilai bilangan asam setiap 2 jam. VCOH yang mempunyai nilai bilangan asam yang optimal, selanjutnya diuji aktivitas antibakterinya terhadap Salmonella secara in-vitrodilakukan dengan metode difusi agar menggunakan pencadang kertas (6 mm, Oxoid) dengan mengamati zona hambat. Zona hambat yang paling baik, selanjutnya digunakan untuk pengujian aktivitas antibakteri VCOH terhadap Salmonella secara in-vivo pada mencit galur mus musculus yang diinfeksikan dengan S. typhimurium, diamati tanda-tanda klinis dan dihitung jumlah koloni dalam feses mencit serta perubahan histopatologi jaringan hati dan limpa.Aktivitas antibakteri bahan uji dibandingkan dengan kontrol positif, negatif dan pembanding kloramfenikol. Hasil pengujian dianalisis statistik dengan analisis variansi (ANAVA; α0,05) dan dilanjutkan dengan uji Post HocDuncan untuk melihat perbedaan nilai rata-rata signifikan antar kelompok perlakuan.
Nilai bilangan asam yang konstan diperoleh pada masa inkubasi setelah 14 jam. Hasil hidrolisis enzimatis VCO menunjukkan bahwa waktu hidrolisis optimal adalah 14 jam. Hasil pengujian aktivitas antibakteri VCOH terhadap S. typhi (ATCC 00786) dan S. typhimurium (ATCC 14028) secara in-vitro menunjukkan
adanya daya hambat dengan peningkatan konsentrasi. Konsentrasi 1000 μl/ml
menunjukkan daya hambat paling baik 13,900±0,3033 mm terhadap S. typhi dan 14,333±0,6623 mm terhadap S. typhimurium. Secara in-vivo, mencit sebelumnya diinfeksi S.typhimurium kemudiandiberi bahan uji.Mencit yang dipajankan dengan bahan uji menunjukkan penurunan jumlah koloni bakteri dalam feses mencit. Tanda klinis dan kerusakan histologis jarang dijumpai pada mencit yang diberi bahan uji, sedangkankelompok yang tidak diobati dan kelompok pembanding menunjukkan adanya tanda-tanda klinis diare, berat badan menurun dan kerusakan histopatologis pada hati dan limpa. Dapat disimpulkan bahwa hasil hidrolisis enzimatis VCO memiliki potensi sebagai anti-Salmonella.
Kata kunci: antibakteri, minyak kelapa murni (VCO), enzimatis,Salmonella
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IN-VITRO AND IN-VIVO ANTIBACTERIAL ACTIVITY OF ENZYMATIC HYDROLYSIS OF VIRGIN COCONUT OILAGAINST
SALMONELLA TYPHI AND SALMONELLA TYPHIMURIUM
Abstract
Salmonellosis is a zoonotic disease caused by pathogenic bacteria Salmonellawhich the route of infection is usually through cross-contamination of food (foodborne disease). Treatment of Salmonellosis commonly used antibiotics in addition to correcting dehydration and electrolyte disturbances. Virgin Coconut Oil (VCO) has the medical potential such as antibacterial, antiviral, and antifungal activity of its medium chain fatty acids (MCFAs), particularly lauric acid (C12: 0) in its monoglycerides form (monolaurin or ML). The aim of this study was to examine the in-vitro and in-vivo antibacterial activity of the enzymatic hydrolysis of VCO (VCOH)which generates a combination of lauric acid and monolaurin against Salmonellawhich are expected to provide a synergistic effect.
The sample of VCOthatin this study wasPalem Mustika VCO-Virgin Coconut Oil, produced by Siti Nurbaya, West Sumatra. VCO was hydrolyzed by enzyme lipase (Lipozyme® TL IM) for 18 hours with interval observation time of free fatty acids value every 2 hours.VCOH which has the optimum free fatty acid value, then investigated the in-vitro antibacterial activity of VCOH against Salmonellaby diffusion agar method using the paper discs (6 mm, Oxoid)by observing the zone of inhibition.The best inhibitory zone, then used for investigated in-vivo antibacterial activity of VCOHagainst Salmonella was examined in a S. typhimurium infection mice mus musculus model, clinical signs were observed and counted the number of colonies S. typhimurium from feces and histopathological changes in liver and spleen tissue.Antibacterial activity of VCOHwas compared with the positive control, negative control and comparison of chloramphenicol. Data were analyzed statistically with analysis of variance
(ANOVA; α 0,05), followed by Duncan's Post Hoc test to see the difference in the average value significantly between treatment groups.
The fatty acids value was obtained constant after incubation for 14 hours. The result of enzymatic hydrolysis VCO show that the optimum hydrolysis time was 14 hours. The in-vitroantibacterial activity of VCOHagainst S.typhi (ATCC 00786) andS.typhimurium (ATCC 14028) was found to have inhibition with increased concentration. The concentration of1000μl/ml showed the best inhibition of the 13.900 ± 0.3033 mm to S.typhi and 14.333 ± 0.6623 mm to S.typhimurium. In in-vivo, mice were initially infected with S. typhimurium and then withVCOH. The feces of miceshowed a decrease in the number of colonies S. typhimurium in the feces. Clinical signs and histological damage were rarely observed in treated mice, whereas the untreated group and the comparison group showed clinical signs of diarrhea, weight loss and histological damage in the liver and spleen. It can be concluded that the results of this study indicate that the enzymatic hydrolysis of VCO has potential as an anti-Salmonella.
Key word: antibacterial, Virgin Coconut Oil (VCO), enzymatic,Salmonella
