Skip to main content
Log in

Evaluation of the probiotic characteristics of Bacillus species isolated from different food sources

  • Original Article
  • Published:
Annals of Microbiology Aims and scope Submit manuscript

Abstract

Probiotics have established their efficacy as dietary adjuncts providing benefits to consumers. However, selection of probiotics before incorporation into diet requires close scrutiny in the form of in vitro as well as in vivo tests. Three bacteriocinogenic Bacillus sp., namely, B. licheniformis Me1, B. flexus Hk1, and B. subtilis Bn1 previously isolated from milk, cheese and fermented beans, respectively, were characterized for typical in vitro probiotic criteria. When compared to probiotic Bacillus coagulans, all three cultures were found to possess better acid and bile tolerance. Cultures Me1 and Bn1, except Hk1, showed bile salt hydrolase activity. A marked difference in adhesion to hydrocarbons and auto-aggregation properties from 10–80 and 60–99%, respectively, were observed for the tested cultures. Highest antioxidant activity was measured for culture Hk1 (66.6%), whereas least activity of 53% was observed for culture Bn1. Cultures Me1 and Bn1 were sensitive to all the antibiotics tested, whereas Hk1 and B. coagulans showed resistance to the penicillin group of β-lactum antibiotics. All the tested cultures showed a broad spectrum of activity against food-borne pathogens. In co-cultivation studies, B. licheniformis Me1 completely inhibited the growth of the indicator pathogen Listeria monocytogenes ScottA. Overall, the test cultures exhibiting potential probiotic characteristics, particularly B. licheniformis Me1, can serve as probiotics of commercial interest.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Abriouel H, Franz CMAP, Omar NB, Galvez A (2011) Diversity and applications of Bacillus bacteriocins. FEMS Microbiol Rev 35:201–232

    Article  PubMed  CAS  Google Scholar 

  • Bae HD, Yanke LJ, Cheng KJ, Selinger LB (1999) A novel staining method for detecting phytase activity. J Microbiol Methods 39:17–22

    Article  PubMed  CAS  Google Scholar 

  • Beaumont M (2002) Flavouring composition prepared by fermentation with Bacillus sp. Int J Food Microbiol 75:189–196

    Article  PubMed  CAS  Google Scholar 

  • Begley M, Hill C, Gahan CGM (2006) Bile salt hydrolase activity in probiotics. Appl Environ Microbiol 72:1729–1738

    Article  PubMed  CAS  Google Scholar 

  • Bernhard K, Schrempf H, Goebel W (1978) Bacteriocin and antibiotic resistance plasmids in Bacillus cereus and Bacillus subtilis. J Bacteriol 133:897–903

    PubMed  CAS  Google Scholar 

  • Brashears MM, Galyean ML, Loneragan GH, Killinger-Mann K (2003) Prevalence of Escherichia coli O157, H7 and performance by beef feedlot cattle given Lactobacillus direct-fed microbials. J Food Prot 66:748–754

    PubMed  CAS  Google Scholar 

  • Caldini G, Trotta F, Cenci G (2002) Inhibition of 4-nitroquinoline-1-oxide genotoxicity by Bacillus strains. Res Microbiol 153:165–171

    Article  PubMed  CAS  Google Scholar 

  • Canzi E, Guglielmetti S, Mora D, Tamagnini I, Parini C (2005) Conditions affecting cell surface properties of human intestinal Bifidobacteria. Antonie van Leeuwenhoek 88:207–219

    Article  PubMed  CAS  Google Scholar 

  • Cartman M, Ragione L, Woodward MJ (2007) Bacterial spore formers as probiotics for poultry. Food Sci Technol Bull 4:21–30

    Google Scholar 

  • Chateau N, Deschamps AM, Hadj-Sassi A (1994) Heterogeneity of bile salts resistance in the Lactobacillus isolates from a probiotic consortium. Lett Appl Microbiol 18:42–44

    Article  Google Scholar 

  • Chou L, Weimer B (1999) Isolation and characterization of acid- and bile-tolerant isolates from strains of Lactobacillus acidophilus. J Dairy Sci 82:23–31

    Article  PubMed  CAS  Google Scholar 

  • Collado MC, Grześkowiak L, Salminen S (2007) Probiotic strains and their combination inhibit in vitro adhesion of pathogens to pig intestinal mucosa. Curr Microbiol 55:260–265

    Article  PubMed  CAS  Google Scholar 

  • Conway PL, Gorbach SL, Goldin BR (1987) Survival of lactic acid bacteria in the human stomach and adhesion to intestinal cells. J Dairy Sci 70:1–12

    Article  PubMed  CAS  Google Scholar 

  • Cosson C, Deschamps AM (1994) Behavior of probiotic strains in the presence of bile and bile salts. Nutr Food Microbiol 12:93–98

    Google Scholar 

  • Cotelle N, Bemier JL, Catteau JP, Pommery J, Wallet JC, Gaydou EM (1996) Antioxidant properties of hydroxyl flavones. Free Radical Biol Med 20:35–43

    Article  CAS  Google Scholar 

  • Cutting SM (2011) Bacillus probiotics. Food Microbiol 28:214–220

    Article  PubMed  Google Scholar 

  • De Vecchi E, Drago L (2006) Lactobacillus sporogenes or Bacillus coagulans: misidentification or mislabelling? Int J Probio Prebio 1:3–10

    Google Scholar 

  • Duan XJ, Zhang WW, Li XM, Wang BG (2006) Evaluation of antioxidant property of extract and fractions obtained from a red alga, Polysiphonia urceolata. Food Chem 95:37–43

    Article  CAS  Google Scholar 

  • Dunne C, O’Mahony L, Murphy L, Thornton G, Morrissey D, O’Halloran S, Feeney M, Flynn S, Fitzgerald G, Daly C, Kiely B, O’Sullivan GC, Shanahan F, Collins JK (2001) In vitro selection criteria for probiotic bacteria of human origin: correlation with in vivo findings. Am J Clin Nutr 73:386S–392S

    PubMed  CAS  Google Scholar 

  • Durkee DL (2010) Coming out of the dairy case: New developments in shelf stable probiotic foods. http://www.foodmaster.com

  • EUCAST (2011). Breakpoint tables for interpretation of MICs and zone diameters. Version 1.3, January 2011. Available at http://www.eucast.org/clinical_breakpoints/. Accessed: March 18, 2011

  • FAO/WHO (2006) Probiotics in food. Health and nutritional properties and guidelines for evaluation. FAO Food and Nutrition Paper 85, ftp://ftp.fao.org/docrep/fao/009/a0512e/a0512e00.pdf

  • Fuller R (1989) Probiotics in man and animals. J Appl Bacteriol 66:365–378

    Article  PubMed  CAS  Google Scholar 

  • Ganeshan P, Kumar CS, Bhaker N (2008) Antioxident properties of methanol extract and its solvent fractions obtained from selected Indian red seaweeds. Bioresource Technol 99:2717–2723

    Article  Google Scholar 

  • Gilbert P, Evans DJ, Evans E, Duguid IG, Brown MRW (1991) Surface characteristics and adhesion of Escherichia coli and Staphylococcus epidermidis. J Appl Bacteriol 71:72–77

    PubMed  CAS  Google Scholar 

  • Gilliland SE, Staley TE, Bush LJ (1984) Importance in bile tolerance of Lactobacillus acidophilus used as a diatery adjunct. J Dairy Sci 67:3045–3051

    Article  PubMed  CAS  Google Scholar 

  • Gracheva NM, Gavrilov AF, Soloveva AI, Smirnov VV, Sorokulova IB, Reznik SR, Chudnovskaia NV (1996) The efficacy of the new bacterial preparation biosporin in treating acute intestinal infections. Zh Mikrobiol Epidemiol Immunobiol 1:75–77

    PubMed  Google Scholar 

  • Guo XH, Kim JM, Nam HM, Park SY, Kim JM (2010) Screening lactic acid bacteria from swine origins for multistrain probiotics based on in vitro functional properties. Anaerobe 16:321–326

    Article  PubMed  Google Scholar 

  • Havenaar R, Huis in’t Veld JHJ (1992) Probiotics: a general view. The lactic acid bacteria. In: Wood BJB (ed) The lactic acid bacteria in health and disease, vol 1. Elsevier, London, pp 151–71

    Google Scholar 

  • Hong HA, Duc LH, Cutting SM (2005) The use of bacterial spore formers as probiotics. FEMS Microbiol Rev 29:813–835

    Article  PubMed  CAS  Google Scholar 

  • Hong HA, Huang JM, Khaneja R, Hiep LV, Urdaci MC, Cutting SM (2008) The safety of Bacillus subtilis and Bacillus indicus as food probiotics. J Appl Microbiol 105:510–520

    Article  PubMed  CAS  Google Scholar 

  • Hosoi T, Kiuchi, K (2004) Production and probiotic effects of natto. In: Ricca E, Henriques AO, Cutting SM (eds) Bacterial spore formers: probiotics and emerging applications. Horizon Bioscience, Norfolk, pp 143–154.

  • Hyronimus B, Le Marrec C, Hadj Sassi A, Deschamps A (2000) Acid and bile tolerance of spore-forming lactic acid bacteria. Int J Food Microbiol 61:193–197

    Article  PubMed  CAS  Google Scholar 

  • Inatsu Y, Nakamura N, Yuriko Y, Fushimi T, Watanasiritum L, Kawamoto S (2006) Characterization of Bacillus subtilis strains in Thua nao, a traditional fermented soybean food in northern Thailand. Lett Appl Microbiol 43:237–242

    Article  PubMed  CAS  Google Scholar 

  • Kim KP, Rhee CH, Park HD (2002) Degradation of cholesterol by Bacillus subtilis SFF34 isolated from Korean traditional fermented flatfish. Lett Appl Microbiol 35:468–472

    Article  PubMed  CAS  Google Scholar 

  • Lee Y-K, Nomoto K, Salminen S, Gorbach SL (1999) Handbook of Probiotics. Wiley, New York

  • Maruo T, Sakamoto M, Toda T, Benno Y (2006) Monitoring the cell number of Lactococcus lactis subsp. cremoris FC in human feces by real-time PCR with strain-specific primers designed using the RAPD technique. Int J Food Microbiol 110:69–76

    Article  PubMed  CAS  Google Scholar 

  • Mazza P (1994) The use of Bacillus subtilis as an antidiarrhoeal microorganism. Boll Chim Farm 133:3–18

    PubMed  CAS  Google Scholar 

  • Meroni PL, Palmieri R, Barcellini W, De Bartolo G, Zanussi C (1983) Effect of long-term treatment with B. subtilis on the frequency of urinary tract infections in older patients. Chemioterapia 2:142–144

    Google Scholar 

  • Mora D, Fortina MG, Parini C, Ricci G, Gatti M, Giraffa G, Manachini PL (2002) Genetic diversity and technological properties of Streptococcus thermophilus strains isolated from dairy products. J Appl Microbiol 93:278–287

    Article  PubMed  CAS  Google Scholar 

  • Nedelcheva P, Denkova Z, Denev P, Slavchev A, Krastanov A (2010) Probiotic strain Lactobacillus plantarum NBIMCC 2415 with antioxidant activity as a starter culture in the production of dried fermented meat products. Biotech Biotechnological Equip 24:1624–1630

    Article  Google Scholar 

  • Otero MC, Ocana VS, Nader-Macias EM (2004) Bacterial surface characteristics applied to selection of probiotic microorganisms. Methods Mol Biol 268:435–440

    PubMed  Google Scholar 

  • Pereira DIA, mc Cartney AL, Gibson GR (2003) An in vitro study of the probiotic potential of a bile salt hydrolysing Lactobacillus fermentum strain and determination of its cholesterol lowering properties. Appl Environ Microbiol 69:4743–4752

    Article  PubMed  CAS  Google Scholar 

  • Pinchuk IV, Bressollier P, Verneuil B, Fenet B, Sorokulova IB, Megraud F, Urdaci MC (2001) In vitro anti- Helicobacter pylori activity of the probiotic strain Bacillus subtilis is due to secretion of antibiotics. Antimicrob Agents Chemother 45:3156–3161

    Article  PubMed  CAS  Google Scholar 

  • Raghavendra P, Halami PM (2009) Screening, selection and characterization of phytic acid degrading lactic acid bacteria from chicken intestine. Int J Food Microbiol 133:129–134

    Article  PubMed  CAS  Google Scholar 

  • Rolfe RD (2000) The role of probiotic cultures in the control of gastrointestinal health. J Nutr 130:396S–402S

    PubMed  CAS  Google Scholar 

  • Rowland I (1999) Probiotics and benefits to human health-the evidence in favour. Environ Microbiol 1:375–382

    Article  PubMed  CAS  Google Scholar 

  • Salvetti S, Celandroni F, Ghelardi E, Baggiani A, Senesi S (2003) Rapid determination of vitamin B2 secretion by bacteria growing on solid media. J Appl Microbiol 95:1255–1260

    Article  PubMed  CAS  Google Scholar 

  • Sanders ME, Morelli L, Tompkins TA (2003) Sporeformers as human probiotics: Bacillus, Sporolactobacillus and Brevibacillus. Compre Rev Food Sci Food Safe 2:101–110

    Article  Google Scholar 

  • Schillinger U, Guigas C, Holzapfel WH (2005) In vitro adherence and other properties of lactobacilli used in probiotic yogurt-like products. Int Dairy J 15:1289–1297

    Article  CAS  Google Scholar 

  • Sorokulova IB (1996) Outlook for using bacteria of the genus Bacillus for the design of new biopreparations. Antibiot Khimioter 41:13–15

    PubMed  CAS  Google Scholar 

  • Sorokulova IB, Pinchuk IV, Denayrolles M, Osipova IG, Huang JM, Cutting SM, Urdaci MC (2008) The safety of two Bacillus probiotic strains for human use. Dig Dis Sci 53:954–963

    Article  PubMed  Google Scholar 

  • Stecchini ML, Torre MD, Munari M (2001) Determination of peroxy radical scavenging of lactic acid bacteria. Int J Food Microbiol 64:183–188

    Article  PubMed  CAS  Google Scholar 

  • Sumi H, Nakajima N, Yatagai C (1995) A unique strong fibrinolytic enzyme (datsuwokinase) in skipjack “Shiokara”, a Japanese traditional fermented food. Comp Biochem Physiol 112:543–547

    Article  CAS  Google Scholar 

  • Tagg JR, Mac Given AR (1971) Assay system for bacteriocin. J Appl Bacteriol 21:943

    CAS  Google Scholar 

  • Tam NKM, Uyen NQ, Hong HA, Duc LH, Hoa TT, Serra CR, Henriques AO, Cutting SM (2006) The intestinal life cycle of Bacillus subtilis and close relatives. J Bacteriol 188:2692–700

    Article  PubMed  CAS  Google Scholar 

  • Urdaci MC, Pinchuk I (2004) Antimicrobial activity of Bacillus probiotics In: Ricca E, Henriques AO, Cutting SM (eds) Bacterial spore formers: probiotics and emerging applications. Horizon Bioscience, Norfolk, pp 171–182.

  • Verschuere L, Rombaut G, Sorgeloos P, Verstraete W (2000) Probiotic bacteria as biological control agents in aquaculture. Mic Mol Biol Rev 64:655–671

    Article  CAS  Google Scholar 

  • Vijayendra SVN, Gupta RC (2011) Assessment of probiotic and sensory properties of dahi and yoghurt prepared using bulk freeze-dried cultures in buffalo milk. Ann Microbiol. doi:10.1007/s13213-011-0331-5

Download references

Acknowledgements

The authors wish to acknowledge The Director, CFTRI, Mysore and Head, Food Microbiology Department, CFTRI, for providing the facilities. Work was carried out under MLP83. N.V. acknowledges CSIR for the fellowship. We thank Mrs. Vyshnavi BS for her technical involvement in the initial stage of this work and also Dr. Vijayendra SVN for editing the manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Prakash M. Halami.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nithya, V., Halami, P.M. Evaluation of the probiotic characteristics of Bacillus species isolated from different food sources. Ann Microbiol 63, 129–137 (2013). https://doi.org/10.1007/s13213-012-0453-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13213-012-0453-4

Keywords

Navigation