Isolasi dan karakterisasi bakteri lactobacillus spp. dari susu sapi lokal

Kandou, Abigail Sabrina (2022) Isolasi dan karakterisasi bakteri lactobacillus spp. dari susu sapi lokal. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Lactobacillus spp. adalah sekelompok bakteri asam laktat (BAL) yang memiliki potensi sebagai probiotik. Salah satu sumber BAL yang dapat memberi keuntungan bagi kesehatan adalah susu sapi. Penelitian ini bertujuan untuk mengisolasi dan mengkarakterisasi bakteri Lactobacillus spp., terutama Lactobacillus delbrueckii, dari susu sapi lokal. Penelitian ini dilakukan dalam beberapa tahap yaitu: isolasi dan pemurnian isolat, karakterisasi morfologi (koloni dan sel) isolat, karakterisasi aktivitas biokimia (aktivitas katalase, hemolitik, dan fermentasi gula), dan uji ketahanan isolat (garam, pH rendah, dan suhu). Isolasi bakteri bermula dengan inokulasi sampel susu terdilusi pada MRS agar dengan metode spread plate. Koloni dimurnikan dengan metode 3-way streak. Kemudian, karakterisasi isolat dilakukan melalui pengamatan morfologi koloni dan sel. Pengamatan morfologi sel dilakukan dengan pewarnaan Gram, pewarnaan endospora, dan pewarnaan tahan asam. Uji aktivitas biokimia termasuk uji aktivitas katalase, uji aktivitas hemolitik, dan uji fermentasi 9 jenis gula (glukosa, fruktosa, sukrosa, laktosa, sorbitol, galaktosa, maltosa, manosa, dan manitol). Lalu, uji ketahanan isolat dilakukan terhadap NaCl (2%, 4%, dan 6%), pH (2,3,4,5,6, dan 7) dan suhu (7°C, 37°C, dan 45°C). Dari hasil isolasi, terdapat 19 koloni dengan morfologi koloni yang sesuai dengan koloni L. delbrueckii. Jumlah isolat berkurang menjadi 14 setelah morfologi sel diamati. Ke-14 isolat merupakan bakteri berbentuk batang Gram positif yang tidak menghasilkan endospora dan tidak memiliki lapisan lilin pada dinding sel. Uji biokimia menghasilkan 5 isolat yang memiliki profil kemampuan fermentasi gula yang sesuai dengan L. delbrueckii subsp. lactis (isolat R, P, dan N), Lactobacillus delbrueckii subsp. indicus (isolat M), dan Lactobacillus delbrueckii subsp. bulgaricus (isolat L). Isolat R, P, N, dan M bersifat gamma hemolitik sedangkan isolat L bersifat alpha hemolitik. Uji ketahanan garam mengungkapkan bahwa kelima isolat kandidat bakteri Lactobacillus delbrueckii bersifat toleran terhadap NaCl 1-4% dan sensitif terhadap NaCl 6%. Hanya isolat R dan P tidak dapat tumbuh pada pH 2 dan 3 sedangkan isolat N, M, dan L masih dapat tumbuh walaupun sangat sedikit. Semua isolat tumbuh pada 37°C dan 45°C tetapi tidak pada suhu 7°C. Hasil 16S rRNA sequencing mengungkapkan isolat L adalah bakteri Limosilactobacillus fermentum./Lactobacillus spp. are a group of lactic acid bacteria (LAB) that has great potential as probiotics. Cow milk is a great source of LAB. This study was aimed to isolate and characterize Lactobacillus spp., especially Lactobacillus delbrueckii, that was obtained from local cow milk. This study was conducted in several general phases which include: isolate isolation and purification, isolate morphological characterization (colony and cell), biochemical characterization (catalase, hemolytic, and sugar fermentation capabilities), and tolerance tests (salt, low pH, and temperature). Isolation began with the inoculation of the milk sample onto MRS agar using the spread plate method. The colonies were purified using the 3- way streak method. Isolate characterization was then done through colony and cell observation. Biochemical characterization included catalase test, hemolytic test and sugar fermentation test comprising of 9 sugars (glucose, fructose, sucrose, lactose, sorbitol, galactose, maltose, mannose, dan mannitol). Tolerance tests were then conducted against NaCl (2%, 4%, and 6%), pH (2,3,4,5,6, and 7) and temperature (7°C, 37°C, and 45°C). The isolation yielded 19 colonies matching the criteria of an L. delbrueckii colony. The number of isolates decreased to 14 after cell morphology was observed. Only 14 out of 19 isolates were Gram positive rods that did not produce endospores and did not have a waxy layer covering its cell wall. Biochemical tests revealed only 5 isolates had a sugar fermentation capability profile that matched with L. delbrueckii, specifically L. delbrueckii subsp. lactis (isolate R, P, and N), Lactobacillus delbrueckii subsp. indicus (isolate M), and Lactobacillus delbrueckii subsp. bulgaricus (isolate L). Isolate R, P, N, and M displayed gamma hemolytic capabilities whilst isolate L displayed alpha hemolytic capability. Tolerance tests revealed that all five of Lactobacillus delbrueckii isolate candidates were tolerant to NaCl 1-4% but growth was suppressed at NaCl 6%. Only isolate R and P were not tolerant to pH 2 and 3 whilst isolate N, M, and L still grew, albeit very weak. All the isolates grew at 37°C and 45°C but not at 7°C. 16S rRNA sequencing revealed isolate L as Limosilactobacillus fermentum.

Item Type: Thesis (Bachelor)
Creators:
CreatorsNIMEmail
Kandou, Abigail Sabrina01113180031abigail.sabrina2@gmail.com
Contributors:
ContributionContributorsNIDN/NIDKEmail
Thesis advisorJo, JuandyUNSPECIFIEDUNSPECIFIED
Thesis advisorSugata, MarceliaUNSPECIFIEDUNSPECIFIED
Uncontrolled Keywords: Isolasi, Karakterisasi, Probiotik, Lactobacillus delbreuckii, Bakteri Asam Laktat
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: University Subject > Current > Faculty/School - UPH Karawaci > Faculty of Science and Technology > Biology
Current > Faculty/School - UPH Karawaci > Faculty of Science and Technology > Biology
Depositing User: Abigail Sabrina Kandou
Date Deposited: 25 Aug 2022 05:50
Last Modified: 25 Aug 2022 05:50
URI: http://repository.uph.edu/id/eprint/49871

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