ISSN 2348-313X (Print) International Journal of Life Sciences Research ISSN 2348-3148 (online) Vol. 8, Issue 3, pp: (1-7), Month: July - September 2020, Available at: www.researchpublish.com
Isolation of Probiotic Lactobacillus Salivarius from Traditional Cow Curd Chenchu Lakshmi Kandati, Pramoda Kumari J* Dept of Microbiology, S. V. University Tirupati, AP, India
Abstract: A probiotic is a live microbial food supplement which beneficially affects the host animal by improving its intestinal microbial balance. In the present study, we isolated probiotic Lactobacillus salivarius from cow curd. Pour plate technique was used to isolate the organisms. Samples were used directly and also diluted to 10-1, 10-2 and 10-3 using sterile peptone water. 1 ml aliquot of the samples and dilutions were plated into MRS (Man, Rogosa and Sharpe) agar (Oxoid LTD, Basingstoke, England) selective medium according to Dave and Shah,. The plates were incubated at 37°C for 3 days under aerobic conditions. All isolates were sub-cultured by streaking method to get pure cultures. Pure culture of specific probiotic isolate from the sample was identified on the basis of biochemical tests such as catalase, oxidase and IMVIC (indole, methyl-red, voges proskaur, citrate utilization) tests. Gram-staining was carried out. Identification of isolates was carried out based on the method described by Sakazaki and Shimad Collins et al., (1989). The identified Lactobacillus species was confirmed through PCR reaction using genus specific primer. The curd has suggested as a source of potential probiotic strain which has probiotic properties. Keywords: Lactobacillus, probiotics, biochemical tests, rRNA sequence.
I. INTRODUCTION Probiotics are commonly defined as mono or mixed cultures of live microbes that when applied to an animal or human posses a beneficial effect on the health of host by improving the balance of the indigenous microflora, and also improved nutrition, growth and prevention of various gastrointestinal disorders. (R Fuller 1999). Probiotics are widely used to prepare fermented dairy products like curd, yogurt, and these are important source of probiotic Lactobacillus (Kamrunnahar Islam et al., 2015). These are a group of bacteria that are frequently used as probiotics. They have a long history in their use as probiotics in the food industry and the Lactobacillus strains are “generally recognized as safe” (GRAS). They are commonly found in the environment such as soil, water, decaying plant materials, as well as in the normal microflora of the gastrointestinal tract (GIT) of animals ( Kizerwetter – Swida and Binek, 2005). L Salivarius found in the mouth and small intestine has antimicrobial activities and breaks down proteinase and produces vitamin B, enzymes and Lactic acid. L salivarius helps to inhibit Salmonella and ulcer Lactobacillus causing by bacteria H. Pylori. Lactobacillus salivarius belongs to the terrabacteria group in the class Firmicutes. It is in the class Bacilli, of the order Lactobacillales, and belongs to the family Lactobacillus (“European Bioinformatics” 2018). Lactobacillus salivarius is a well characterized, rod-shaped Gram-positive species of probiotic bacteria (Chapot-Chartier et al., 2014). L salivarius is characterized as a lactic acid bacterium (LAB), which denotes its ability to produce lactic acid by fermentation (Kuratsu et al., 2010).
II. MATERIALS AND METHODS Cow curd sample was collected from the local area. Pour plate technique was used to isolate the organisms. Samples were used directly and also diluted to 10-1, 10-2 , 10-3, using sterile peptone water. 1 ml aliquot of the samples and dilutions were plated into MRS (Man, Rogosa and Sharpe) agar (Oxoid LTD, Basingstoke, England) according to Dave and selective medium (Shah, 1996). The plates were incubated at 37°C for 3 days under aerobic conditions. All isolates were sub-cultured by streaking method to get pure cultures.
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