Identifikasi dan karakterisasi isolat SU-KC1a dari air susu ibu = Identification and characterization SU-KC1a isolate from human breast milk

Kim, Youngchae (2021) Identifikasi dan karakterisasi isolat SU-KC1a dari air susu ibu = Identification and characterization SU-KC1a isolate from human breast milk. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Telah diketahui bahwa bifidobakteria dan Lactobacillus sp. termasuk L. plantarum merupakan probiotik yang dapat diisolasi dari air susu ibu (ASI). Penelitian ini bertujuan untuk mengidentifikasi serta mengkarakterisasi isolat SU-KC1a, SU-KC2, dan SU-KC3 yang berasal dari ASI, dan mengevaluasi potensi sebagai probiotik. Metode pada penelitian ini terdiri dari: pemurnian isolat menggunakan media mTPY (mupirocin-trypticase phytone yeast extract) secara anaerobik obligat pada suhu 37 oC selama 72 jam; identifikasi morfologi dengan pewarnaan Gram dan acid-fast, uji aktivitas enzim katalase dengan menggunakan H2O2 (3%) dan uji motilitas agar pada tabung agar mTPY; serta uji potensi sebagai probiotik berupa uji ketahanan garam empedu dengan konsentrasi 0,5% - 2% pada media agar mTPY, uji kerentanan terhadap 13 antibiotik dengan metode paper disc dan uji aktivitas antimikroba dengan metode well-diffusion. Identifikasi molekular dilakukan dengan sequencing 16S-rRNA dan dianalisis menggunakan BLAST. Berdasarkan hasil yang didapatkan, ketiga isolat memiliki karakterisik Gram-positif, acid-fast negatif, morfologi sel yang basil pendek, non-motile, dan katalase negatif. Uji potensi probiotik ketiga isolat menunjukkan tahan pada garam empedu 0,5% - 2%, kerentanan pada 7 antibiotik (ampicillin, amoxicillin, erthromycin, chloramphenicol, rifampin, dan neomycin) dan 6 resisten (bacitracin, vancomycin, streptomycin, kanamycin, Nalidix acid, dan tetracycline). Uji aktivitas antimikroba menunjukkan penghambatan terhadap E. coli dan S. aureus. Berdasarkan hasil identifikasi molekular 16S rRNA, isolat SU-KC1a, SU-KC2 dan SU-KC3 merupakan strain yang identik, yaitu Lactiplantibacillus plantarum. Dapat disimpulkan, bahwa isolat SU-KC1a berpotensi sebagai probiotik, namun masih diperlukan penelitian lebih lanjut untuk digunakan sebagai probiotik./ It is known that bifidobacteria and Lactobacillus sp. such as L. plantarum are probiotics which can be isolated from human breast milks. This study aims to identify and characterize SU-KC1a, SU-KC2, and SU-KC3 isolates derived from breast milk, and evaluate their potential as probiotics. The methods in this study consisted of: purification of isolates using mTPY (mupirocin-trypticase phytone yeast extract) media in obligate anaerobic way at 37 oC for 72 hours; morphological identification with Gram and acid-fast staining, test for catalase enzyme activity using H2O2 (3%) and agar motilation test on mTPY agar tubes; the probiotic potential test were evaluated through bile salt resistance test with concentration of 0.5% - 2% on mTPY agar media, the susceptibility test to 13 antibiotics using the paper disc method and the antimicrobial activity test using the well-diffusion method. Molecular identification was performed by 16S-rRNA sequencing and analyzed using BLAST. Based on the results obtained, the three isolates were Gram-positive, acid-fast negative, cell morphology with short bacilli, non-motile, and catalase negative. The evaluation of potential probiotic of the three isolates showed resistance to 0.5%-2% bile salts, susceptibility to 7 antibiotics (ampicillin, amoxicillin, erthromycin, chloramphenicol, rifampin, and neomycin) and 6 resistance (bacitracin, vancomycin, streptomycin, kanamycin, Nalidix acid and tetracycline). Antimicrobial activity test showed a zone of inhibition to E. coli and S. aureus. Based on the results of the molecular identification of 16S rRNA, SU-KC1a, SU-KC2 and SU-KC3 isolates were identical strains, which is Lactiplantibacillus plantarum. Therefore, it can be concluded that the SU-KC1a isolate can potentially be used as a probiotic. However, further research is required to support this claim.

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