Fasiha Shah et al. Int. Res. J. Pharm. 2013, 4 (11) Page 40 INTERNATIONAL RESEARCH JOURNAL OF PHARMACY www.irjponline.com ISSN 2230 – 8407 Research Article ISOLATION AND FREEZE DRYING OF CERTAIN PROBIOTIC STRAINS Fasiha Shah 1 *, Kahlid Idrees Khan 2 and Kausar Tasneem Jaffery 3 1 Rak Medical and Health Sciences University, UAE 2 Punjab University Lahore, Pakistan 3 Vetenary Research Institute Lahore, Pakistan *Corresponding Author Email: cookiepharmacist@yahoo.com Article Received on: 30/09/13 Revised on: 31/10/13 Approved for publication: 11/11/13 DOI: 10.7897/2230-8407.041110 IRJP is an official publication of Moksha Publishing House. Website: www.mokshaph.com © All rights reserved. ABSTRACT The aim of the present study was to evaluate few probiotics strains available commercially in Lahore city and isolation and identification of strains from these commercial probiotics. The identified and isolated probiotic strains were freeze dried. These strains included Lactobacillus acidophilus, Bifidumbacterium bifidus and Saccharomyces boulradii. Both the commercial and laboratory freeze dried probiotic micro-organisms were subjected to microbial studies. The isolation of the microbes was done by using different types of growth media and identification of microbes was based on fermentation of different carbohydrates, gas production from glucose, growth at various temperature and tolerance to sodium chloride, colony characteristics and staining. The actual microbial count in laboratory of commercial probiotics indicated that there was a discrepancy in the viable count stated on the label of Lactobacillus acidophilus, Bifidumbacterium bifidus i.e., the actual viable count is less than the stated viable count on the label. This laboratory evidence suggest that bacteria cannot survive longer time on shelves which is probably the main reason that probiotics inspite of their in vivo effectiveness have not found a good way in physician prescription pattern. However, such discrepancy was not observed in case of yeast. The microbiological studies suggested that it is feasible to isolate, identify and freeze dry the microbes in the laboratory and pilot plant scale. The microbial count after freeze drying of laboratory prepared probiotics had shown that anaerobic bacteria (Bifidumbacterium bifidus) survived more during the freeze drying as compared to Lactobacillus acidophilus and yeast survive more than the both bacterial strains. Keywords: Probiotics, fermentation and freeze drying. INTRODUCTION As defined by the Food and Agriculture Organization of the United Nations and the World Health Organization in October, 2001, probiotics "are live microorganisms which when administered in adequate amounts confer a health benefit on the host". The word Probiotic was coined in the 1960s to name substances produced by microorganisms which promoted the growth of other microorganisms 1 . The probiotic bacteria used in commercial products today are mainly members of the genera Lactobacillus, Bifidobacterium and Yeast but other microbes are also used. Lactobacillus species from which probiotic strains have been isolated include are L. acidophilus BG2FO4, INT-9, NCFB 1748 NCFM and DDS-1, L. plantarum ST31, L. johnsonii LA1, L. casei Shirota, L. fermentum and L. rhamnosus strain such as GG 60 .Specific strains of bifidobacteria used as probiotics include B. breve strain Yakult and RO7O, B. lactis Bb12, B. longum RO23, B. bifidum RO71, B. infantis RO33, B. longum BB536 and SBT-292811; Streptococcus thermophiles a probiotic micro-organism. Streptococcus salivarus subspecies thermophilus type 1131 is another strain used in commercial probiotics 2 . The principal probiotic yeast is Saccharomyces boulardii 3 . Probiotics exhibit antimicrobial, immunomodulatory, anti carcinogenic, anti diarrheal and antioxidant activities. The antimicrobial activity of probiotics is thought to be accounted for, in large part, by their ability to colonize the colon and reinforce the barrier function of the intestinal mucosa. In addition, some probiotics have been found to secrete antimicrobial substances. These substances are known as bacteriocins. Such bacteriocin has been isolated from Lactobacillus plantarum ST31, a probiotic derived from sour dough 4 . Saccharomyces boulardii has been found to secrete a protease which digests two protein exotoxins, toxin A and B, which appear to mediate diarrhea and colitis caused by Clostridium difficile. The proposed uses of probiotics are, in the treatment of various digestive illnesses such as ulcer due to Helicobacter pylori 5 , clearing up constipation 6 , reducing episodes of chronic diarrhea 7 , antibiotic associated diarrhea, alleviation of symptoms of lactose malabsorption 8 , acute diarrhea 9 , candiasis 10 , antibiotic effects such as antibacterial effects against E. coli 11 , Crohn’s disease 12 , liver injury 13 , food poisoning 14 , ulcerative colitis 15 , act as anti hypertensive 16 , reduce the risk of heart attack 17 and reduce cholesterol level in the blood 18 , can prevent urinary tract and vaginal infections, can be used against mucosal infections 19 and antiviral effects such as gastroenteritis in children 20 . Today there are more than 70 lactic acid bacteria containing products worldwide. More than 53 different types of probiotic milk products are marketed in Japan alone, including frozen desserts like frozen yogurt 21 , frozen dough 22 , sour cream and ice cream 23 . Probiotics have been incorporated into drinks e.g., kefir like beverages 24 and whey beverages 25 and effervescent preparations 26 . Those are also being marketed as tablets such as enteric soluble royal jelly tablets 27 , enteric coated tablets 28 and chewable tablets 29 , as capsules such as gel encapsulated probiotics 30,31 and as microcapsules 32 . These have been marketed as mineral water composition 33 . Probiotics are also marketed in combination with antibiotics 34 , vitamins and prebiotics 35 . The Fermented Milks and Lactic Acid Beverages Association in Japan have introduced a standard of a minimum of 107 CFU/ml viable probiotic cells for fresh dairy products (minimum concentration of probiotic required for beneficial effect). The scientist’s have 36 suggested a minimum viable number of 106 CFU/ml or gram but recommends 108 CFU/g to compensate for reduction through passage through the gut. A typical daily dose should supply about 3 to 5 billion live microorganisms 37 . The typical dose of Saccharomyces boulardii yeast is 250 mg twice daily 38 .