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Synergistic effects of probiotic Leuconostoc mesenteroides and Bacillus subtilis in malted ragi (Eleucine corocana) food for antagonistic activity against V. cholerae and other beneficial properties

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Abstract

Finger millet (Elucine corocana), locally known as ragi, and probiotics have been recognized for their health benefits. In the present work we describe novel probiotic ragi malt (functional food) that has been prepared using ragi and probiotic Leuconostoc mesenteroides (Lm) and Bacillus subtilis natto (Bs), alone and in combination, for antagonistic activity against Vibrio cholerae (Vc). In vitro studies using pure cultures showed that each probiotic strain (Lm or Bs) was able to inhibit the planktonic growth of Vc as well as its ability to make biofilms and adhere to extracellular matrix proteins (fibronectin, Fn) that may function in vivo as initial ports of entrance of the pathogen. Interestingly, the combination of both probiotic strains (Lm plus Bs) produced the strongest activity against the Vc. When both cultures were used together in the ragi malt the antimicrobial activity against Vc was enhanced due to synergistic effect of both probiotic strains. The inclusion of both probiotic strains in the functional food produced higher amounts of beneficial fatty acids like linoleic and linolenic acid and increased the mineral content (iron and zinc). The viability and activity of Lm and Bs against Vc was further enhanced with the use of adjuvants like ascorbic acid, tryptone, cysteine hydrochloride and casein hydrolysate in the ragi malt. In sum, the intake of probiotic ragi malt supplemented with Lm and Bs may provide nutrition, energy, compounds of therapeutic importance and antagonistic activity against Vc to a large extent to the consumer.

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References

  • Agrawal R, Rati ER, Vijayendra SVN, Varadaraj MC, Prasad MS, Krishnanand (2000) Flavor profile of idli batter prepared from defined microbial starter cultures. World J Microbiol Biotechnol 16:687–690

    Article  Google Scholar 

  • AOAC (2007) analytical Official methods of analysis. AOAC International 18th ed (985.35 and 968.080 ISBN: 978–0935584783

  • Arabolaza A, Nakamura A, Pedrido ME, Martelotto L, Orsaria L, Grau R (2003) Characterization of a novel inhibitory feedback of the anti-anti-sigma factor SpoIIAA on activation of SpooA transcription factor during development in Bacillus subtilis. Mol Microbiol 47:1251–1263

    Article  CAS  Google Scholar 

  • Barák I, Ricca E, Cutting S (2005) From fundamental studies of sporulation to applied spore research. Mol Microbiol 55:330–338

    Article  Google Scholar 

  • Bassler BL, Losick R (2006) Bacterially speaking. Cell 125:237–246

    Article  CAS  Google Scholar 

  • Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Canadian J Biochem Physiol 37(911):917

    Google Scholar 

  • Bryskier A (1998) Roxithromycin: review of its antimicrobial activity. J Antimicrobial Chemotherapy 41(supplB):1–21

    Article  CAS  Google Scholar 

  • Carvalho AS, Silvaa J, Hob P, Teixeira P, Malcataa FX, Gibbsa P (2004) Relevant factors for the preparation of freeze dried lactic acid bacteria. Int Dairy J 14:835–847

    Article  CAS  Google Scholar 

  • Casula G, Cutting S (2002) Bacillus probiotics: spore germination in the gastrointestinal tract. Appl Environ Microbiol 68:2344–2352

    Article  CAS  Google Scholar 

  • Champevolier D, LeNoc P, Bryskier A, Croize J (1993) Comparative in vitro activity of 9 macrolides against 50 Vibrio cholerae strains. In: Proceedings of the 33rd Interscience Conference on antimicrobial agents and chemotherapy. New Orleans. LA, American Society for Microbiology. Washington, DC. 231:161

  • Chavan UD, Chavan JK, Kadam SS (1989) Effect of fermentation on soluble proteins and in vitro protein digestibility of sorghum, green gram and sorghum green gram blends. J Fd Sci 53:1574–1575

    Article  Google Scholar 

  • Chavarri FJ, De Paz M, Nueez MM (1988) Cryoprotective agents for frozen concentrated starters from non- bitter Streptococcus lactis strains. Biotech Lett 10:11–16

    Article  CAS  Google Scholar 

  • Da Silva SH, Vieira EC, Nicoli JR (1998) Antagonism against Vibrio cholerae by bacterial diffusible compound in the fecal microbiota of rodents. Rev Microbiol 29:3

    Google Scholar 

  • Damiani P, Gobbetti M, Cossignanai L, Corsetti A, Simonetti MS, Rossi J (1996) The sourdough microflora characterization of hetero- and homo fermentative lactic acid bacteria, yeasts and their interactions on the basis of the volatile compounds produced. Lebensm Wiss Technol 29:63–70

    Article  CAS  Google Scholar 

  • Dave RI, Shah NP (1998) Ingredient supplementation effects on viability of probiotic bacteria in yoghurt. Dairy Sci 81:2804–2816

    Article  CAS  Google Scholar 

  • Duc L, Hong H, Barbosa T, Henriques A, Cutting S (2004) Characterization of Bacillus probiotics available for human use. Appl Environ Microbiol 70:2161–2171

    Article  CAS  Google Scholar 

  • Fujita M, Musch MW, Nakagawa YHS, Alverdy J, Kohgo Y, Schneewind O, Jabri B, Chang EB (2007) The Bacillus subtilis quorum-sensing molecule CSF contributes to intestinal homeostasis via OCTN2, a host cell membrane transporter. Cell Host Microbe 4:299–308

    Article  Google Scholar 

  • Fukuda S, Toh H, Hase K, Oshima K, Nakanishi Y, Yoshimura K, Tobe T, Clarke JM, Topping DL, Suzuki T, Taylor TD, Itoh K, Kikuchi J, Morita H, Hattori M, Ohno H (2011) Bifidobacteria can protect from enteropathogenic infection through production of acetate. Nature 469:543–547

    Article  CAS  Google Scholar 

  • Hamon M, Lazazzera B (2001) The sporulation transcription factor SpooA is required for biofilm development in Bacillus subtilis. Mol Microbiol 42:1199–1209

    Article  CAS  Google Scholar 

  • Heinzmann S, Entian K, Stein T (2006) Engineering Bacillus subtilis ATCC 6633 for improved production of the lantibiotic subtilin. Appl Microbiol Biotechnol 69:532–536

    Article  CAS  Google Scholar 

  • Helland MH, Wicklund T, Narvhus JA (2004) Growth and metabolosim of selected strains of probiotic bacteria in maize porridge with added malted barley. Int J Food Microbiol 91:305–313

    Article  CAS  Google Scholar 

  • Hosoi T, Kiuchi K (2004) Production and probiotic effects of natto. In: Ricca E, Henriques AO, Cutting S (eds) Bacterial spore formers: probiotics and emerging applications. Horizon Bioscience, UK, Publisher

    Google Scholar 

  • Ingham K, Brew S, Vaz D, Sauder D, McGavin M (2004) Interaction of Staphylococcus aureus fibronectin-binding protein with fibronectin. J Biol Chem 279:42945–42953

    Article  CAS  Google Scholar 

  • Ismail MM Soliman WSE (2010) Studies on probiotic effects of lactic acid bacteria against Vibrio vulnificus in freshwater prawn Macrobrachium osenbergii. J Amer Sci 6 (12). Available on line http://www.jofamericanscience.org/journals/am-sci/am0612/

  • Kalui CM, Mathara JM, Kutima PM (2010) Probiotic potential of spontaneously fermented cereal based foods. A review 9:2490–2498

    CAS  Google Scholar 

  • Kemperman RA, Bolca S, Roger LC, Vaughan EE (2010) Novel approaches for analyzing gut microbes and dietary polyphenols: challenges and opportunities. Microbiology 156:3224–3231

    Article  CAS  Google Scholar 

  • Lombardía E, Rovetto A, Arabolaza A, Grau R (2006) A Lux-S dependent cell to cell language regulates social behavior and development in Bacillus subtilis. J Bacteriol 188:4442–4452

    Article  Google Scholar 

  • Nishito Y, Osana Y, Hachiya T, Popendorf K, Toyoda A, Fujiyama A, Itaya M, Sakakibara Y (2010) Whole genome assembly of a natto production strain Bacillus subtilis natto from very short read data. BMC Genomis 11:243

    Article  Google Scholar 

  • Nout MJR (1991) Ecology of accelerated natural lactic acid fermentation of sorghum-based infant food formulas. Int J Food Microbiol 12:217–224

    Article  CAS  Google Scholar 

  • Oyewole OB (1997) Lactic fermented foods in Africa and their benefits. Fd Control 8:289–297

    Article  Google Scholar 

  • Parsek MR, Greenberg EP (2005) Sociomicrobiology: the connections between quorum sensing and biofilms. Trends Microbiol 13:27–33

    Article  CAS  Google Scholar 

  • Qiu D, Fujita K, Sakuma Y, Tanka T, Oaci Y, Ohshima H, Tomita M, Itaya M (2004) Comparative analysis of physical maps of tour Bacillus subtilis (natto) genomes. Appl Environ Microbiol 70:6247–6256

    Article  CAS  Google Scholar 

  • Salminen S, Isolauri E, Salminen E (1996) Clinical uses of probiotics for stabilizing the gut mucosal barrier: successful strains and future challenges. Antonie Van Leewenhock 70:347–358

    Article  CAS  Google Scholar 

  • Sánchez B, Urdaci M, Margolles A (2010) Extracellular proteins secreted by probiotic bacteria as mediators of effects that promote mucosa-bacteria interactyions. Microbiology 156:3232–3242

    Article  Google Scholar 

  • Sankara RDS, Deosthala YG (1983) Mineral composition, ionizable iron and soluble zinc in malted grains of pearl millet and ragi. Food Chem 11:217–223

    Article  Google Scholar 

  • Schillinger U, Lucke FK (1989) Antibacterial activity of Lactobacillus sake isolated from meat. Appl Environ Microbiol 55(8):1901–1906

    CAS  Google Scholar 

  • Schwarz-Linek U, Höök M, Potts J (2004) The molecular basis of fibronectin-mediated bacterial adherence to host cells. Mol Microbiol 52:631–641

    Article  CAS  Google Scholar 

  • Senesi S (2004) Bacillus spores as probiotic products for human use, chapter 11. In: Ricca E, Henriques AO, Cutting S (eds) Bacterial spore formers: probiotics and emerging applications. Horizon Bioscience, Norwich, United Kingdom

    Google Scholar 

  • Shobha PR, Agrawal R (2007) Volatile compounds of therapeutic importance produced by Leuconostoc mesenteroides, a native laboratory isolate. Turk J Biol 31:35–40

    CAS  Google Scholar 

  • Styriak I, Laukov A, Fallgren C, Wadstrán T (1999) Binding of extracellular matrix proteins by animal strains of Staphylococcal species. Vet Microbiol 67:99–112

    Article  CAS  Google Scholar 

  • Svanberg U (1995) Lactic acid fermented foods for feeding infants. In: Steinkraus KH (ed) Handbook of indigenous fermented foods. Marcel Dekker, New York, pp 310–347

    Google Scholar 

  • Tettech GL, Sefa-Dedeh SK, Phillips RD, Beuchat LR (2004) Survival and growth of acid adapted and unadapted Shigella flexneri in a traditional fermented Ghanian weaning food as affected by fortification with cowpea. Int J Food Microbiol 90:189–195

    Article  Google Scholar 

  • Thomas C, Versalovic J (2010) Probiotic-host communication. Modulation of signaling pathways in the intestine. Gut Microbes 3:148–163

    Article  Google Scholar 

  • Vidya R, Iyer PR (2010) Antagonistic activity of probiotic organism against Vibrio cholerae and Cryptococcus neoformans. Malaysian J Microbiol 6:41–46

    Google Scholar 

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Authors and Affiliations

  1. Department of Food Microbiology, Central Food Technological Research Institute, Mysore, 570020, India

    B. VidyaLaxme & Renu Agrawal

  2. Department of Microbiology, School of Biochemistry and Pharmacy, Rosario National University-CONICET, Rosario, 2000, Argentina

    A. Rovetto & R. Grau

Authors
  1. B. VidyaLaxme
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  2. A. Rovetto
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  3. R. Grau
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  4. Renu Agrawal
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Correspondence to R. Grau or Renu Agrawal.

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VidyaLaxme, B., Rovetto, A., Grau, R. et al. Synergistic effects of probiotic Leuconostoc mesenteroides and Bacillus subtilis in malted ragi (Eleucine corocana) food for antagonistic activity against V. cholerae and other beneficial properties. J Food Sci Technol 51, 3072–3082 (2014). https://doi.org/10.1007/s13197-012-0834-5

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  • DOI: https://doi.org/10.1007/s13197-012-0834-5

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