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Isolasi dan Identifikasi Bakteri Asam Laktat dari Susu Kambing Sebagai Bakteri Antagonis Listeria monocytogenes dan Escherichia coli Penyebab Foodborne Disease

*Annisa Yulina Susilowati  -  Department of Biology, Universitas Diponegoro, Indonesia
Siti Nur Jannah  -  Departemen Biologi, Fakultas Sains dan Matematika, Universitas Diponegoro, Semarang, Indonesia
Hermin Pancasakti Kusumaningrum orcid scopus  -  Departemen Biologi, Fakultas Sains dan Matematika, Universitas Diponegoro, Semarang, Indonesia
Sulistiani Sulistiani Sulistiani scopus  -  Bidang Mikrobiologi, Pusat Penelitian Biologi LIPI, Cibinong Science Center (CSC), Bogor, Indonesia
Received: 12 Dec 2020; Published: 31 Dec 2022.
Editor(s): Siti Susanti, Ph.D

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Abstract

Listeria monocytogenes dan Escherichia coli merupakan kontaminan pangan berbahaya yang dapat mengakibatkan timbulnya foodborne disease. Mikrobia dengan aktivitas antibakteri seperti Bakteri Asam Laktat (BAL) yang berasal dari susu kambing dapat dijadikan solusi untuk mengatasi permasalahan tersebut. Aktivitas antibakteri BAL disebabkan produksi asam laktat yang dapat menurunkan pH medium dan beberapa senyawa bioaktif seperti asam organik, H2O2, diasetil dan bakteriosin. Penelitian dilakukan untuk mengetahui jenis dan potensi BAL susu kambing sebagai antibakteri Listeria monocytogenes dan Escherichia coli. Metode penelitian meliputi isolasi dan pengamatan morfologi BAL, uji ketahanan isolat BAL terhadap suhu dan pH, pewarnaan Gram, uji katalase (H2O2)uji aktivitas antibakteri BAL, serta identifikasi secara molekuler isolat BAL yang menunjukkan potensi antibakteri. Isolat yang mampu menghambat Listeria monocytogenes adalah isolat 372 dengan diameter zona bening 5,2 mm serta isolat 469 menghambat Escherichia coli dengan zona bening berdiameter 5,8 mm. Identifikasi molekuler menunjukkan kedua isolat memiliki ukuran DNA genom 1500 bp tetapi hanya isolat 372 yang dilanjutkan hingga tahap sekuensing karena menunjukkan aktivitas penghambatan terhadap bakteri patogen lebih baik dibandingkan isolat 469 yang ditandai dengan zona bening terlihat lebih jelas. Berdasarkan analisis hasil sekuensing DNA melalui BLAST dapat diketahui bahwa isolat 372 merupakan Pediococcus pentosaceus strain 1931 dengan kemiripan homologi sebesar 99,72%.

Listeria monocytogenes and Escherichia coli are dangerous food contaminants that can cause foodborne disease. Microbes with antibacterial activity such as Lactic Acid Bacteria (LAB) from goat's milk can be used as a solution to overcome these problems. LAB antibacterial activity is due to the production of lactic acid which can lower the pH of the medium and several bioactive compounds such as organic acids, H2O2, diacetyl, and bacteriocins. The study was conducted to determine the type and potential of LAB goat milk as antibacterial agents for Listeria monocytogenes and Escherichia coli. The research methods include isolation and observation of LAB morphology, resistance test of LAB isolates against temperature and pH, Gram staining, catalase test (H2O2), LAB antibacterial activity test, and molecular identification of LAB isolates that show antibacterial potential. Isolates capable of inhibiting Listeria monocytogenes were isolate 372 with clear zone diameter 5.2 mm and isolate 469 inhibited Escherichia coli with clear zone diameter 5.8 mm. Molecular identification showed that both isolates had a genomic DNA size of 1500 bp but only isolate 372 was continued to sequencing because it showed better inhibitory activity against pathogenic bacteria than isolate 469 which was marked with clearer visible zones. Based on the analysis of the results of DNA sequencing through BLAST, it can be seen that isolate 372 is Pediococcus pentosaceus strain 1931 with homology similarity 99.72%.

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Keywords: BAL; foodborne disease; antibakteri; identifikasi

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