Diversity of lactic acid bacteria isolated from fermented mare's milk products based on PCR-RFLP analysis

International Conference on Green Agro-industry and Bioeconomy

农业科学;工业技术（总论）;经济学

Mulyawati, A.I.^1 ; Jatmiko, Y.D.^1 ; Mustafa, I.^1 ; Ardyati, T.^1 ; Suharjono^1

Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Brawijaya, Malang, Indonesia^1

关键词: Good Manufacturing Practices;    Gram-negative bacteria;    Internal transcribed spacers;    Lactic acid bacteria;    Lactobacillus species;    Restriction fragment length polymorphism analysis;    Saline solutions;    Treatment methods;   

Others  :  https://iopscience.iop.org/article/10.1088/1755-1315/230/1/012104/pdf
DOI  :  10.1088/1755-1315/230/1/012104

来源: IOP

【 摘 要 】

This study aimed to characterize the diversity of lactic acid bacteria (LAB) isolated from fermented mare's milk based on PCR-RFLP. The LAB was isolated from three fermented mare's milk products from three different regions i.e Dompu, Bima, and Sumbawa, Province of West Nusa Tenggara, Indonesia. Fermented milk samples (25 mL) was diluted into 225 mL of saline solution and spread plated onto MRS and M17 agar containing 1 % CaCO3, then incubated at 37 °C for 48 hours. Single colonies showing a clear zone were randomly selected and purified. The DNA was extracted by the heat-treatment method and amplified using an internal transcribed spacer (ITS) and 5.8S region spacer primers. The amplicons of the 5.8S region spacer were used for restriction fragment length polymorphism (RFLP) analysis using HaeIII and HindIII enzymes. The DNA bands were characterized to obtain the LAB clusters. The representative of each LAB cluster was amplified using 16S rDNA primer then sequenced. Total of 41 isolates was grouped into 10 clusters based on the PCR-RFLP analysis. However, only three clusters were identified as LAB from Lactobacillus genus. The Lactobacillus species obtained were L. plantarum and two strains of L. rhamnosus. Four clusters were identified as Staphylococcus genus and three others were Ochrobactrum genus. Thus, the LAB isolates from fermented mare milk obtained were the member of Lactobacillus genus. However, the Gram-negative bacteria and Staphylococcus were still found in the fermented products indicating the poor quality of the products. Therefore, a good manufacturing practice should be implemented during the product preparation.