Skip to main content
Log in

Isolation and Characterization of Putative Probiotic Bacterial Strain, Bacillus amyloliquefaciens, from North East Himalayan Soil Based on In Vitro and In Vivo Functional Properties

  • Published:
Probiotics and Antimicrobial Proteins Aims and scope Submit manuscript

Abstract

This study is aimed to isolate some novel probiotics from the soils of North East Himalayas. Eleven Gram positive isolates were obtained in MRS with Oxgall media from soil samples. Four of the isolates withstood the in vitro gastric juice pH 3.0 and 0.45% bile salt tolerance screening. Among these, PBT 3 showed high cell surface hydrophobicity and adhered to the Caco-2 cells. 16s rDNA gene sequences of this probiotic strains were identified as Bacillus amyloliquefaciens (accession no: JF836079). In in vivo bioefficacy evaluation by DSS-induced colitis animals, B. amyloliquefaciens significantly ameliorated the loss in body weight. Further, the treatment altered the levels of myeloperoxidase, lipoperoxides, and mucous content in the colon tissues compared with normal colon. It reduced the protein and mRNA levels of pro-inflammatory cytokines such as TNF-α and IL-1β. These biochemical findings were supported by histopathological evidences. Our study reports the use of B. amyloliquefaciens isolated from Himalayan soil as probiotic and its beneficial effect on IBD for the first time and suggests that this could be used as potential probiotics in functional foods or as a curative agent.

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
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Bhakta JN, Ohnishi K, Munekage Y, Iwasaki K (2010) Isolation and probiotic characterization of arsenic-resistant lactic acid bacteria for uptaking arsenic. World Academy of Science. Eng Technol 71:470–471

    Google Scholar 

  2. Blandizzi C, Natale G, Gherardi G, Lazzeri G, Marveggio C, Colucci R, Carignani D, Del Tacca M (1999) Acid-independent gastroprotective effects of lansoprazole in experimental mucosal injury. Dig Dis Sci 44:2039–2050

    Article  CAS  Google Scholar 

  3. Bobin-Dubigeon C, Collin X, Grimaud N, Robert JM, Le Baut G, Petit JY (2001) Effects of tumor necrosis factor-synthesis inhibitors on rat trinitrobenzene sulphonic acid-induced chronic colitis. Eur J Pharmacol 421:103–110

    Article  Google Scholar 

  4. Cetinkaya A, Bulbuloglu E, Kantarceken B, Ciralik H, Kurutas EB, Buyukbese MA, Gumusalan Y (2006) Effects of L-carnitine on oxidant/antioxidant status in acetic acid-induced colitis. Dig Dis Sci 51:488–494

    Article  CAS  Google Scholar 

  5. Chen CC, Walker WA (2005) Probiotics and prebiotics: role in clinical disease states. Adv Pediatr 52:77–113

    Article  Google Scholar 

  6. Deniz M, Cetinel S, Kurtel H (2004) Blood flow alterations in tnbs-induced colitis: role of endothelin receptors. Inflamm Res 53:329–336

    Article  CAS  Google Scholar 

  7. EFSA (2004) Opinion of the scientific panel on additives and products or substances used in animal feed on a request from the Commission on the safety of the product “Kemzyme W Dry” regarding the ability of Bacillus strains involved in the manufacture to produce toxins. EFSA J 118:1–4

    Google Scholar 

  8. EFSA (2007) European food safety authority. Introduction of a qualified presumption of safety (QPS) approach for assessment of selected microorganisms referred to EFSA opinion of the scientific committee (Question No EFSA-Q-2005–293). Adopted on 19 November 2007. EFSA J 587:1–16

    Google Scholar 

  9. EFSA (2008) Scientific opinion of the panel on additives and products or substances used in animal feed (FEEDAP) on a request from the European Commission on the safety and efficacy of Ecobiol® (Bacillus amyloliquefaciens) as feed additive for chickens for fattening. EFSA J 773:1–13

    Google Scholar 

  10. Einerhand AW, Renes IB, Makkink MK, Vander Sluis M, Buller HA, Dekker J (2002) Role of mucins in inflammatory bowel disease: important lessons from experimental models. Eur J Gastroenterol Hepatol 14:757–765

    Article  CAS  Google Scholar 

  11. Gaggia F, Mattarelli P, Biavati B (2010) Probiotics and prebiotics in animal feeding for safe food production. Int J Food Microbiol 141:S15–S28

    Article  Google Scholar 

  12. Gionchetti P, Rizzello F, Helwig U, Venturi A, Lammers KM, Brigidi P, Vitali B, Poggioli G, Miglioli M, Campieri M (2003) Prophylaxis of pouchitis onset with probiotic therapy: a double-blind, placebo-controlled trial. Gastroenterology 124:1202–1209

    Article  Google Scholar 

  13. Goldin BR, Gorbach SL, Saxelin M, Barakat S, Gualtieri L, Salminen S (1992) Survival of Lactobacillus species (strain GG) in the human gastrointestinal tract. Dig Dis Sci 37:121

    Article  CAS  Google Scholar 

  14. Grisham MB, Beniot JN, Granger DN (1990) Assessment of leukocyte involvement during ischemia and reperfusion of intestine. Methods Enzymol 186:729–742

    Article  CAS  Google Scholar 

  15. Guarnera F, Schaafsmab GJ (1998) Probiotics. Int J Food Microbiol 39:237–238

    Article  Google Scholar 

  16. Havenaar R, Brink NG, Huis In’t Ved JHJ (1992) Selection of strains for probiotics use. In: Fuller R (ed) Probiotics, the scientific basis. Chapman & Hall, London, pp 210–224

    Google Scholar 

  17. Immerseel FV, Ducatelle R, De Vos M, Boon N, Wiele TVD, Verbeke K, Rutgeerts P, Sas B, Louis P, Flint HJ (2010) Butyric acid-producing anaerobic bacteria as a novel probiotic treatment approach for inflammatory bowel disease. J Med Microbiol 59:141–143

    Article  Google Scholar 

  18. Ishikawa H, Akedo I, Umesaki Y et al (2003) Randomized controlled trial of the effect of Bifidobacteria-fermented milk on ulcerative colitis. J Am Coll Nutr 22:56–63

    Google Scholar 

  19. Kim YG, Moon JT, Lee KM et al (2006) The effects of probiotics on symptoms of irritable bowel syndrome. Korean J Gastroenterol 47:413–419

    Google Scholar 

  20. Kos B, Suskovic J, Vukovic S, Simpraga M, Frece J, Matosic S (2003) Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J Appl Microbiol 94:981–987

    Article  CAS  Google Scholar 

  21. Kravtsov EG, Yermolayev AV, Anokhina IV, Yashina NV, Chesnokova VL, Dalin MV (2008) Adhesion characteristics of Lactobacillus is a criterion of the probiotic choice. Bull Exp Biol Med 145:232–234

    Article  CAS  Google Scholar 

  22. Kruis W, Schutz E, Fric P et al (1997) Double-blind comparison of an oral Escherichia coli preparation and mesalazine in maintaining remission of ulcerative colitis. Aliment Pharmacol Ther 11:853–858

    Article  CAS  Google Scholar 

  23. Madsen KL, Doyle JS, Jewell LD, Tavernini MM, Fedorak RN (1999) Lactobacillus species prevents colitis in interleukin 10 gene-deficient mice. Gastroenterology 116:1107–1114

    Article  CAS  Google Scholar 

  24. Makharia GK, Sood A, Midha V (2008) 701 A randomised, double blind, placebo controlled trial of a probiotic preparation, VSL#3, for the treatment of mild to moderate active ulcerative colitis. Gastroenterology 134:A-99

    Article  Google Scholar 

  25. Markert M, Andrews PC, Babior BM (1984) Measurement of O2 production by human neutrophils. The preparation and assay of NADPH oxidase-containing particles from human neutrophils. Methods Enzymol 105:358–365

    Article  CAS  Google Scholar 

  26. Mayra-Makinen A, Manninen M, Gyllenberg H (1983) The adherence of lactic acid bacteria to the columnar epithelial cells of pigs and calves. J Appl Bacteriol 55:241–245

    Article  CAS  Google Scholar 

  27. Mileti E, Matteoli G, Iliev ID, Rescigno M (2009) Comparison of the immunomodulatory properties of three probiotic strains of Lactobacilli using complex culture systems: prediction for in vivo efficacy. PLoS One 4:7056

    Article  Google Scholar 

  28. Ng SC, Plamondon S, Al-Hassi HO et al (2008) Effective probiotic treatment (VSL#3), but not placebo, in acute ulcerative colitis is associated with downregulation of inflammatory intestinal dendritic cells. Gut 57:96

    Google Scholar 

  29. Pelletier C, Bouley C, Cayuela C, Bouttier S, Bourlioux P, Bellon-Fontaine MN (1997) Cell surface characteristics of Lactobacillus casei subsp. casei, Lactobacillus paracasei subsp. paracasei, and Lactobacillus rhamnosus strains. Appl Environ Microbiol 63:1725–1731

    CAS  Google Scholar 

  30. Phelps RJ, McKillip JL (2002) Enterotoxin production in natural isolates of Bacillaceae outside the Bacillus cereus group. Appl Environ Microbiol 68:3147–3151

    Article  CAS  Google Scholar 

  31. Prakash S, Urbanska AM (2008) Colon-targeted delivery of live bacterial cell biotherapeutics including microencapsulated live bacterial cells. Biologics 2:355–378

    CAS  Google Scholar 

  32. Prasad J, Gill H, Smart J, Gopal PK (1998) Selection and characterization of Lactobacillus and Bifidobacterium strains for use as probiotics. Int Dairy J 8:993–1002

    Article  Google Scholar 

  33. Prisciandaro LMG, Butler R, Cummins A, Howarth G (2009) Probiotics and their derivatives as treatments for inflammatory bowel disease. Inflamm Bowel Dis 15:1906–1914

    Article  Google Scholar 

  34. Ramon MA, Villegas I, Sanchez-Hidalgo M, Catalina AL (2006) The effects of resveratrol, a phytoalexin derived from red wines, on chronic inflammation induced in an experimentally induced colitis model. Br J Pharmacol 147:873–885

    Article  Google Scholar 

  35. Rochat T, Bermudez-Humaran L, Gratadoux J, Fourage C, Hoebler C, Corthier G, Langella P (2007) Anti-inflammatory effects of Lactobacillus casei BL23 producing or not a manganese-dependant catalase on DSS-induced colitis in mice. Microb Cell Fact 6:22

    Article  Google Scholar 

  36. Saavedra JM, Bauman NA, oung L, Perman JA, Yolken RH (1994) Feeding of Bifidobacterium and Streptococcus thermophilus to infants in hospital to prevent diarrhea and shedding of rotavirus. Lancet 344:1046–1049

    Article  CAS  Google Scholar 

  37. Sartor RB (1994) Cytokines in intestinal inflammation: pathophysiological and clinical consideration. Gastroenterology 106:533–539

    CAS  Google Scholar 

  38. Sonia M, Harshavardhan K (2011) Gut microbiota is not modified by randomized, double-blind, placebo-controlled trial of VSL#3 in diarrhea-predominant irritable bowel syndrome. Probiotics Antimicrob Proteins 3:1–7

    Article  Google Scholar 

  39. Succi M, Tremonte P, Reale A, Sorrentino E, Grazia L, Pacifico S, Coppola R (2005) Bile salt and acid tolerance of Lactobacillus rhamnosus strains isolated from Parmigiano Reggiano cheese. FEMS Microbiol 244:129–137

    Article  CAS  Google Scholar 

  40. Torres MI, Garci′A-Martin M, Gil A, Rios A (1999) Experimental colitis induced by trinitrobenzenesulphonic acid. An ultrastructural and histochemical study. Digest Dis Sci 44:2523–2529

    Article  CAS  Google Scholar 

  41. Ueda M, Mozaffar S, Tanaka A (1990) Catalase from Candida boidinii 2201. Methods Enzymol 188:463–467

    Article  CAS  Google Scholar 

  42. Vesterlund S, Paltta J, Karp M, Ouwehand AC (2005) Measurement of bacterial adhesion: in vitro evaluation of different methods. J Microbiol Methods 60:225–233

    Article  CAS  Google Scholar 

  43. Whorwell PJ, Altringer L, Morel J et al (2006) Efficacy of an encapsulated probiotic Bifidobacterium infantis 35624 in women with irritable bowel syndrome. Am J Gastroenterol 101:1581–1590

    Article  Google Scholar 

  44. Xu L, Yang ZL, Li P, Zhou YQ (2009) Modulating effect of hesperidin on experimental murine colitis induced by dextran sulfate sodium. Phytomedicine 16:989–995

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful to Entomology research institute for financial assistance. The authors are thankful to the reviewers for their useful comments and feedback.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to S. Ignacimuthu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hairul Islam, V.I., Prakash Babu, N., Pandikumar, P. et al. Isolation and Characterization of Putative Probiotic Bacterial Strain, Bacillus amyloliquefaciens, from North East Himalayan Soil Based on In Vitro and In Vivo Functional Properties. Probiotics & Antimicro. Prot. 3, 175–185 (2011). https://doi.org/10.1007/s12602-011-9081-8

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12602-011-9081-8

Keywords

Navigation