Identification of Bifidobacterium animalis, Bifidobacterium adolescentis and Bifidobacterium bifidum from Stool of Children and Detection of Their Antibacterial Properties

Mansour Amin, Ahmad Farajzadeh Sheikh, Hamed Goodarzi, Mehdi Sormeh

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Department of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences (AJUMS), Ahvaz, Iran.

DOI: 10.4236/aid.2013.33029                

Abstract

Introduction and Objective: The genus Bifidobacterium can generally be found in quantity in the habitats such as human and animal gastrointestinal tract, dental caries, vagina and oral cavity. The aim of this study was to isolate Bifidobacterium from stool and determine their inhibitory effect against some pathogens. Materials and Methods: 130 samples were collected by wet swabs and kept in sterile tubes containing MRS broth media. And Bifidobacterium isolated from stool was enriched in Man-Rogosa-Sharpe medium (MRS) broth and isolated by growing on MRS agar medium and characterized by phenotypic characteristics and PCR technique at genus and species levels. The antimicrobial substance was extracted from ethyl acetate solvent and the antimicrobial activity against some pathogenic bacteria, such as Salmonella typhi and Shigella sonnei, were investigated. Results: Eleven Bifidobacterium bifidum and four Bifidobacterium adolescentis, which were isolated from fresh stool, were identified by PCR. Antimicrobial substance from MRS broth medium was extracted. This antimicrobial compound showed a potent inhibitory activity against four tested bacteria. These bacteria produced acetic acid and lactic acid as inhibitory substances that were different from bacteriocins. Conclusion: Fresh stool may be used as a source of antimicrobial lactic acids bacteria, Bifidobacterium bifidum and adolescentis as two probiotics can establish themselves in gut and urogenital tract to prevent the human body from adverse effects of pathogens.

Keywords

Bifidobacterium; PCR; Antimicrobial Substance; Stool

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	B. Biavati, M. Vescovo, S. Torriani and V. Bottazzi, “Bifidobacteria History, Ecology, Physiology and Applications,” Annals of Microbiology, Vol. 50, No. 2, 2000, pp. 117-131.
[2]	E. S. Klaassens, W. M. de Vos and E. E. Vaughan, “Molecular Approaches to Assess Activity and Functionality of Commensal and Ingested Bifidobacteria in the Human Intestinal Tract,” In: M. Saarela, Ed., Functional Dairy Products (Volume 2), Woodhead Publishing Limited, Cambridge, 2007, 153-159.
[3]	S. Anuradha and K. Rajeshwan, “Probiotics in Health and Disease,” JIACM, Vol. 6, No. 1, 2005, pp. 67-72.
[4]	S. Salminen, “Human Studies on Probiotics: Aspects of Scientific Documentation,” Scandinavian Journal of Nutrition, Vol. 45, 2001, pp. 8-14.
[5]	K. Harish and T. Varghese, “Probiotics in Humans— Evidence Based Review,” Calicut Medical Journal, Vol. 4, No. 4, 2006, p. e3.
[6]	M.-T. Liong, “Roles of Probiotics and Prebiotics in Colon Cancer Prevention: Postulated Mechanisms and In-Vivo Evidence,” International Journal of Molecular Sciences, Vol. 9, No. 5, 2008, pp. 854-863. doi:10.3390/ijms9050854
[7]	G. Marelli, E. Papaleo and A. Ferrari, “Lactobacilli for Prevention of Urogenital Infection: A Review,” European Review for Medical and Pharmacological Sciences, Vol. 8, No. 2, 2004, pp. 87-95.
[8]	M. E. Falagas, G. I. Betsi and S. Athanasiou, “Prebiotics for Prevention of Recurrent Vulvovaginal Candidiasis: A Review,” Journal of Antimicrobial Chemotherapy, Vol. 58, No. 2, 2006, pp. 266-272. doi:10.1093/jac/dkl246
[9]	B. Biavati, M. Vescovo, S. Torriani and V. Bottazzi, “Bifidobacteria: History, Ecology, Physiology and Applications,” Annals of Microbiology, Vol. 50, No. 2, 2000, pp. 117-131.
[10]	E. V. Mahmoud, L. O. Svensson, S. E. Olsson and A. Mardh, “Antichlamydial Activity of Vaginal Secretion,” American Journal of Obstetrics & Gynecology, Vol. 172, No. 4, 1995, pp. 1268-1272. doi:10.1016/0002-9378(95)91491-9
[11]	S. Garg, R. A. Anderson, J. Chany, D. P. Waller, X. H. Diao, K. Vermani and L. J. Zaneveld, “Properties of a New Acid-Buffering Bioadhesive Vaginal Formulation (ACID-FORM),” Contraception, Vol. 64, No. 1, 2001, pp. 67-75. doi:10.1016/S0010-7824(01)00217-7
[12]	W. M. De Vos and J. Hugenholtz, “Engineering Metabolic Highways in Lactococci and Other Lactic Acid Bacteria,” Trends in Biotechnology, Vol. 22, No. 2, 2004, pp. 72-79. doi:10.1016/j.tibtech.2003.11.011
[13]	B. A. Forbes, D. F. Sahm and A. S. Welssfeld, “Baily and Scotts Diagnostic Microbiology,” 12th Edition, Mosby Inc., St. Louis, 2007, pp. 109-214.
[14]	D. Roy and S. Sirois, “Molecular Differentiation of Bifidobacterium Species with Amplified Ribosomal DNA Restriction Analysis and Alignment of Short Regions, of the ldh Gene,” FEMS Microbiology Letters, Vol. 181, No. 1, 2000, pp. 17-24.
[15]	T. Matsuki, K. Watanabe, R. Tanaka and H. Oyaizu, “Rapid Identification of Human Intestinal Bifidobacteria by 16S rRNA-Targeted Species- and Group-Specific Primers,” FEMS Microbiology Letters, Vol. 167, No. 2, 1998, pp. 113-121. doi:10.1111/j.1574-6968.1998.tb13216.x
[16]	R. F. Wang, W. W. Cao and C. E. Cerniglia, “PCR Detection and Quantitation of Predominant Anaerobic Bacteria in Human and Animal Fecal Samples,” Applied and Environmental Microbiology, Vol. 62, No. 4, 1996, pp. 1242-1247.
[17]	M. Amin and B. P. Kapadnis, “Heat Stable Antimicrobial Activity of Allium ascalonicum against Bacteria and Fungi,” Indian Journal of Experimental Biology, Vol. 43, No. 8, 2005, pp. 751-754.
[18]	I. V. Chervinec, V. M. Bondarenco, H. A. Shabanova, A. M. Samoukina and V. N. Chervinec, “Bacteriocinogenic High Antagonistic Strains of Lactobacillus,” Zhurnal Mikrobiologii Epidemiologii i Immunobiologii, Vol. 7, 2006, pp. 79-81.
[19]	E. Kheadr, N. Dabour, C. Le Lay, C. Lacroix and I. Fliss, “Antibiotic Susceptibility Profile of Bifidobacteria as Affected by Oxgall, Acid, and Hydrogen Peroxide Stress,” Antimicrobial Agents and Chemotherapy, Vol. 51, No. 1, 2007, pp. 169-174. doi:10.1128/AAC.00261-06
[20]	M. Shuhaimi, A. M. Yazid, A. M. Ali, M. H. Ghazali, H. Zaitum and N. A. Nur Atiqah, “Antibacterial Activity, Antimicrobial Susceptibility and Adherence Properties of Bifidobacterium infantis G4,” Pakistan Journal of Biological Sciences, Vol. 2, No. 4, 1999, pp. 1231-1235. doi:10.3923/pjbs.1999.1231.1235
[21]	S. Kozhakhmetov, S. S. Oralbayeva, A. R. Kushugulova, K. Almagambetov, A. K. Abzhalelov and E. M. Ramankulov, “Creation of the Probiotic Consortium on the Base of Strains of Bifidobacterium spp.,” Malaysian Journal of Microbiology, Vol. 5, No. 2, 2009, pp. 67-72.
[22]	S. A. Ibrahim, S. R. Dharmavaram, C. W. Seo and G. Shahbazi, “Antimicrobial Activity of Bifidobacterium Longum (NCFB 2259) as Influenced by Spices Internet,” Journal of Food Safety, Vol. 2, 2001, pp. 6-8.