PROBIOTIC BACTERIA AS YOGHURT STARTER AND ITS IMPLICATION EFFECT TO THE PATHOGENIC AND NON PATHOGENIC BACTERIA IN MICE GASTROINTESTINAL

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1 PROBIOTIC BACTERIA AS YOGHURT STARTER AND ITS IMPLICATION EFFECT TO THE PATHOGENIC AND NON PATHOGENIC BACTERIA IN MICE GASTROINTESTINAL Lovita Adriani, Hendronoto A.W. Lengkey Faculty of Animal Husbandry, University of Padjadjaran, Bandung, INDONESIA The purpose of the research was to study the effect of bacteria consortium of Lactobacillus bulgaricus, Streptococcus thermophillus, Lactobacillus acidophilus and Bifidobacteria on the ecosystem of gastrointestinal in mice.the aim of this study was to explore the differences between yoghurt content from different consortium, with 1,25% dosage; in mice during three until five weeks. Also, the effect on the number of population of non pathogenic bacteria (Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus and Bifidobacteria ) and the total pathogenic bacteria in the segment of the mice gut (jejunum, ilium and colon). Results indicated that the bacteria mixture have a good implementation in microbial intestine of mice, which increased the population of non pathogenic and decreased of pathogenic bacteria. Keywords: total numbers of bacteria, pathogenic bacteria, and non pathogenic bacteria. According to the previous studies, some genera of lactic acid bacteria and bifidobacteria make an extremely important group of probiotic bacteria. Microflora of the gastrointestinal tract of human or animal, they offer considerable potensial as probiotic because of their history of save use and the general body of evidence that supports their positives role (Björkstén, et al, 2001, Guarner and Malagelada, 2003a, Sears,2005 and Steinhoff, 2005). Namely probiotics are microorganism which had been included in food without any adverse effects and which were present in the gastrointestinal tract for health. At present, these microorganism, called probiotics, have been selected from mostly lactic acid bacteria, e.g. Lactobacillus acidophilus. Bifidobacteria is a part of the normal intestinal microflora of human, since the microorganism are indigenous to the colon. The importance of an indigenous microflora in the gastrointestinal tract as a natural resistance factor against potential pathogenic microorganism was already recognised by Metchnicoff. Probiotic strain can be used only, if the microorganism active in the body of the host if the fulfill a large number of criteria. On the other side, lactic acid and acetic acid caused intestine acidity and can prevented the growth of pathogenic bacteria. Those acids reduce absorbsion of ammonia and amine since the large number of ammonia and amine can rise blood pressure, cholesterol,and cancer because of nitrosamine. Lactobacillus is a group of gram positive anaerobic colonic bacteria that produce lactic acid. Supplementation with Lactobacillus and the resultant increased colonic levels of this organism has maintained the health of subjects with several intestinal disorders including diarrhea, ulcerative colitis (Bettelheim, et al, 1974 and Tap, et al, 2009). Research has shown that Lactobacillus effectively competes with pathogenic bacteria in the colon for binding to the epithelial cells that line the intestines (Schwiertz, 2003). It also inhibits pathogenic bacteria by producing lactic acid and increasing epithelial mucous production. 262

2 MATERIALS AND METHODS Lucrări Științifice vol 53 seria Medicină Veterinară Raw milk used for making yoghurt, from farm animal at the faculty. The bacteria are pure cultivated Bifidobacterium, Lactobacillus acidophilus, Lactobacillus bulgaricus and Streptococcus thermophilus. The identification of the cultures was based on the characteristics of lactobacilli and streptococci as described in Bergey s Manual of Determinative Bacteriology (Holt et al, 1994). 120 mice, from a commercial hatchery. The experimental design were Completely Randomized Design (CRD), with six treatment and four replication. Every cages was filled with five mice. RESULTS AND DISCUSSION Total non pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal (cfu/ml). In Table 1, presents the total non pathogenic bacteria in each segment of 6 weeks mice gastro intestinal (cfu/ml). In Table 1, the total of non pathogenic bacteria in the colon are higher than the total bacteria in the stomach, jejunum and ileum, especially with ration + Bifidobacteria starter (R3, R4 and R5). Helpful bacteria prevent the growth of pathogenic species by competing for nutrition and attachment sites to the epithelium of the colon. Symbiotic bacteria are more at home in this ecological niche and are thus more successful in the competition. Indigenous gut floras also produce bacteriocins which are proteinacious toxins that inhibit growth of similar bacterial strains, substances which kill harmful microbes and the levels of which can be regulated by enzymes produced by the host (Guarner and Malagelada, 2003a). The resident gut microflora positively control the intestinal epithelial cell differentiation and proliferation through the production of short chain fatty acids. They also mediate other metabolic effects such as the syntheses of vitamins like biotin and folate as well as absorption of ions including Magnesium, Calcium and Iron (O'Hara and Shanahan, 2006). 263

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4 Universitatea de Științe Agricole și Medicină Veterinară Iași Notes : Lb = Lactobacillus bulgaricus, La = Lactobacillus acidophilus R0 = Controle ration (without yoghurt) R2 = Controle ration + Lb, St and La starter R4 = Controle ration + Lb, St, La and B starter St = Streptococcus thermophilus, B = Bifidobacterium spp. R1 = Controle ration + Lb and St starter R3 = Controle ration + Lb, St and B starter R5 = Controle ration + La and B starter Total pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal (cfu/ml). In Table 2, presents the total pathogenic bacteria in each segment of 6 weeks mice gastro intestinal (cfu/ml). Tabel 2. Total pathogenic bacteria in each segment of 6 weeks Mice Gastro Intestinal Notes : B = Bacillus, Mcc = Micrococci, Stp = Staphylococcus aureus. R0 = Controle ration (without yoghurt) R1 = Controle ration + Lb and St starter R2 = Controle ration + Lb, St and La starter R3 = Controle ration + Lb, St and B starter R4 = Controle ration + Lb, St, La and B starter R5 = Controle ration + La and B starter From the data from Table 2, the mice that have been feed with R5, has reducing the pathogenic bacteria especially in the colon. This fact are agree with Beaugerie and Petit (2004) that fermentation process, since it produces lactic acid and different fatty acids, also serves to lower the ph in the colon, preventing the proliferation of harmful species of bacteria and facilitating that of helpful species. The ph may also enhance the excretion of carcinogens. Metabolic function The gut flora plays a major role in metabolizing dietary carcinogens (Junjie Qin; et al (2009); the microcomponents and the macrocomponents. The microcomponents are genotoxic and the major focus is on recent advances in heterocyclic amines (HCAs) which are produced by cooking proteinaceous food such as meat and fish which can then induce tumors in organs like breast, colon and prostate. HCAs are naturally occurring therefore the complete avoidance ofthem is impractical which is why the metabolic function of gut flora of such components is of 265

5 Lucrări Științifice vol 53 seria Medicină Veterinară great importance to our body as this would help in prevention of such tumors that are difficult to avoid. The macrocomponents consists of the excessive intake of fat and sodium chloride which can later promote tumors such as in breasts and colons from fat and gastric carcinogenesis from sodium chloride(guarner and Malagelada, 2003b) CONCLUSION The yoghurt with consortium starters with Lactobacillus acidophilus and Bifidobacterium spp. with 1.25% dosage of mice body weight have been raised the non pathogenic population and decreased the pathogenic bacteria in colon. BIBLIOGRAPHY 1. Beaugerie L, Petit JC (2004). "Microbial gut interactions in health and disease. Antibiotic associated diarrhoea". Best Pract Res Clin Gastroenterol 18 (2): doi: /j.bpg PMID Bettelheim KA, Breadon A, Faiers MC, O'Farrell SM, Shooter RA (1974). "The origin of O serotypes of Escherichia coli in babies after normal delivery". J Hyg (Lond) 72 (1): PMID Björkstén B, Sepp E, Julge K, Voor T, Mikelsaar M (2001). "Allergy development and the intestinal microflora during the first year of life". J. Allergy Clin. Immunol. 108 (4): doi: /mai PMID (01) Guarner F, Malagelada JR (2003a). "Gut flora in health and disease". Lancet 361 (9356): doi: /s (03) PMID (03) Guarner F, Malagelada JR (2003b). "Role of bacteria in experimental colitis". Best Pract Res Clin Gastroenterol 17 (5): doi: /s (03) PMID Holt, J.G., N.R. Krieg, P.H.A. Sneath, J. T. Staley, and S. T. Williams, Bergey s Manual of determination Bacteriology, 9th ed. Williams and Williams. Baltimore. p Junjie Qin; et al (2009), "A human gut microbial gene catalogue established by metagenomic sequencing", Nature (464): 59 65, doi: /nature08821, retrieved O'Hara AM, Shanahan F (2006). "The gut flora as a forgotten organ". EMBO Rep. 7 (7): doi: /sj.embor PMID Schwiertz A, Gruhl B, Löbnitz M, Michel P, Radke M, Blaut M (2003). "Development of the intestinal bacterial composition in hospitalized preterm infants in comparison with breast fed, full term infants". Pediatr. Res. 54 (3): doi: /01.pdr a. PMID core/templatejournal/ lwwgateway/media/landingpage.htm?issn= &volume=54&issue=3&spage= Sears CL (2005). "A dynamic partnership: celebrating our gut flora". Anaerobe 11 (5): doi: /j.anaerobe PMID (05) Steinhoff U (2005). "Who controls the crowd? New findings and old questions about the intestinal microflora". Immunol. Lett. 99 (1): doi: /j.imlet PMID (05) Tap J, Mondot S, Levenez F, Pelletier E, Caron C, Furet JP, Ugarte E, Muñoz Tamayo R, Paslier DL, Nalin R, Dore J, Leclerc M (2009). "Towards the human intestinal microbiota phylogenetic core.". Env. Microbiol. 11 (10): doi: /j x. PMID