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Relationships between the genome and some phenotypical properties of Lactobacillus fermentum CECT 5716, a probiotic strain isolated from human milk

  • Applied genetics and molecular biotechnology
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Abstract

Lactobacillus fermentum CECT 5716, isolated from human milk, has immunomodulatory, anti-inflammatory, and anti-infectious properties, as revealed by several in vitro and in vivo assays, which suggests a strong potential as a probiotic strain. In this work, some phenotypic properties of L. fermentum CECT 5716 were evaluated, and the genetic basis for the obtained results was searched for in the strain genome. L. fermentum CECT 5716 does not contain plasmids and showed neither bacteriocin nor biogenic amine biosynthesis ability but was able to produce organic acids, glutathione, riboflavin, and folates and to moderately stimulate the maturation of mouse dendritic cells. No prophages could be induced, and the strain was sensitive to all antibiotics proposed by European Food Safety Authority (EFSA) standards, while no transmissible genes potentially involved in antibiotic resistance were detected in its genome. Globally, there was an agreement between the phenotype properties of L. fermentum CECT 5716 and the genetic information contained in its genome.

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References

  • Ardite E, Sans M, Panes J, Romero FJ, Pique JM, Fernandez-Checa JC (2000) Replenishment of glutathione levels improves mucosal function in experimental colitis. Lab Invest 80:735–744

    Article  CAS  PubMed  Google Scholar 

  • Arribas B, Rodriguez-Cabezas ME, Comalada M, Bailon E, Camuesco D, Olivares M, Xaus J, Zarzuelo A, Gálvez J (2009) Evaluation of the preventative effects exerted by Lactobacillus fermentum in an experimental model of septic shock induced in mice. Br J Nutr 101:2–4

    Article  Google Scholar 

  • Asrar FM, O’Connor DL (2005) Bacterially synthesized folate and supplemental folic acid are absorbed across the large intestine of piglets. J Nutr Biochem 16:587–593

    Article  CAS  PubMed  Google Scholar 

  • Bañuelos O, Fernández L, Corral JM, Valdivieso-Ugarte M, Adrio JL, Velasco J (2008) Metabolism of prebiotic products containing beta(2–1) fructan mixtures by two Lactobacillus strains. Anaerobe 14:184–189

    Article  PubMed  Google Scholar 

  • Bover-Cid S, Holzapfel WH (1999) Improved screening procedure for biogenic amine production by lactic acid bacteria. Int J Food Microbiol 53:33–41

    Article  CAS  PubMed  Google Scholar 

  • Camuesco D, Comalada M, Rodriguez-Cabezas ME, Nieto A, Lorente MD, Concha A, Zarzuelo A, Galvez J (2004) The intestinal anti-inflammatory effect of quercitrin is associated with an inhibition in iNOS expression. Br J Pharmacol 143:908–918

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Cárdenas N, Calzada J, Peirotén A, Jiménez E, Escudero R, Rodríguez JM, Medina M, Fernández L (2014) Development of a potential probiotic fresh cheese using two Lactobacillus salivarius strains isolated from human milk. Biomed Res Int. doi:10.1155/2014/801918

    PubMed Central  PubMed  Google Scholar 

  • Church FC, Swaisgood HE, Porter DH, Catignain GL (1983) Spectrophotometric assay using o-phthaldialdehyde for determination of proteolysis in milk and isolated milk proteins. J Dairy Sci 66:1219–1227

    Article  CAS  Google Scholar 

  • Connell SR, Tracz DM, Nierhaus KH, Taylor DE (2003) Ribosomal protection proteins and their mechanism of tetracycline resistance. Antimicrob Agents Chemother 47:3675–3681

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Cueva C, Moreno-Arribas MV, Martín-Alvarez PJ, Bills G, Vicente MF, Basilio A, Rivas CL, Requena T, Rodríguez JM, Bartolomé B (2010) Antimicrobial activity of phenolic acids against commensal, probiotic and pathogenic bacteria. Res Microbiol 161:372–382

    Article  CAS  PubMed  Google Scholar 

  • Danielsen M, Wind A (2003) Susceptibility of Lactobacillus spp. to antimicrobial agents. Int J Food Microbiol 82:1–11

    Article  CAS  PubMed  Google Scholar 

  • Díaz-Ropero M, Martin R, Sierra S, Lara-Villoslada F, Rodriguez JM, Xaus J, Olivares M (2006) Two Lactobacillus strains, isolated from breast milk, differently modulate the immune response. J Appl Microbiol 102:337–343

    Google Scholar 

  • Dudeja PK, Kode A, Alnounou M, Tyagi S, Torania S, Subramanian VS, Said HM (2001) Mechanism of folate transport across the human colonic basolateral membrane. Am J Physiol Gastrointest Liver Physiol 281:G54–60

    CAS  PubMed  Google Scholar 

  • EFSA (2012) Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA J 10:2740

    Google Scholar 

  • Egervarn M, Danielsen M, Roos S, Lindmark H, Lindgren S (2007) Antibiotic susceptibility profiles of Lactobacillus reuteri and Lactobacillus fermentum. J Food Prot 70:412–418

    PubMed  Google Scholar 

  • Fernández L, Langa S, Martín V, Maldonado A, Jiménez E, Martín R, Rodríguez JM (2013) The human milk microbiota: origin and potential roles in health and disease. Pharmacol Res 69:1–10

    Article  PubMed  Google Scholar 

  • Gil-Campos M, López MÁ, Rodriguez-Benítez MV, Romero J, Roncero I, Linares MD, Maldonado J, López-Huertas E, Berwind R, Ritzenthaler KL, Navas V, Sierra C, Sempere L, Geerlings A, Maldonado-Lobón JA, Valero AD, Lara-Villoslada F, Olivares M (2012) Lactobacillus fermentum CECT 5716 is safe and well tolerated in infants of 1–6 months of age: a randomized controlled trial. Pharmacol Res 65:231–238

    Article  PubMed  Google Scholar 

  • Gonzalez R, Sanchez de Medina F, Galvez J, Rodriguez-Cabezas ME, Duarte J, Zarzuelo A (2001) Dietary vitamin E supplementation protects the rat large intestine from experimental inflammation. Int J Vitam Nutr Res 71:243–250

    Article  CAS  PubMed  Google Scholar 

  • Grover S, Sharma VK, Mallapa RH, Batish VK (2013) Draft genome sequence of Lactobacillus fermentum Lf1, an Indian isolate of human gut origin. Genome Announc 1:e00883–13

    PubMed Central  PubMed  Google Scholar 

  • Horwitz W (2000) Official methods of analysis of AOAC international. AOAC International, Gaithersburg

    Google Scholar 

  • Jayashree S, Rajendhran J, Jayaraman K, Kalaichelvan G, Gunasekaran P (2011) Improvement of riboflavin production by Lactobacillus fermentum isolated from yogurt. Food Biotechnol 25:240–251

    Article  CAS  Google Scholar 

  • Jeurink PV, van Bergenhenegouwen J, Jiménez E, Knippels LM, Fernández L, Garssen J, Knol J, Rodríguez JM, Martín R (2013) Human milk: a source of more life than we imagine. Ben Microbes 4:17–30

    Article  CAS  Google Scholar 

  • Jiménez E, Langa S, Martín V, Arroyo R, Martín R, Fernández L, Rodríguez JM (2010) Complete genome sequence of Lactobacillus fermentum CECT 5716, a probiotic strain isolated from human milk. J Bacteriol 192:4800

    Article  PubMed Central  PubMed  Google Scholar 

  • Juarez del Valle M, Laiño J, Savoy de Giori G, LeBlanc JG (2014) Use of lactic acid bacteria as a biotechnological strategy to increase riboflavin levels in soymilk. Food Res Int 62:1015–1019

    Article  CAS  Google Scholar 

  • Katla A-K, Kruse H, Johnsen G, Herikstad H (2001) Antimicrobial susceptibility of starter culture bacteria used in Norwegian dairy products. Int J Food Microbiol 67:147e52

    Article  Google Scholar 

  • Krause LJ, Forsberg CW, O’Connor DL (1996) Feeding human milk to rats increases Bifidobacterium in the cecum and colon which correlates with enhanced folate status. J Nutr 126:1505–1511

    CAS  PubMed  Google Scholar 

  • Kullisaar T, Zilmer M, Mikelsaar M, Vihalemm T, Annuk H, Kairane C, Kilk A (2002) Two antioxidative lactobacilli strains as promising probiotics. Int J Food Microbiol 72:215–224

    Article  CAS  PubMed  Google Scholar 

  • Laiño JE, LeBlanc JG, Savoy de Giori GS (2012) Selection of folate producing starter cultures of yogurt isolated from Northwestern Argentina. Can J Microbiol 58:581–588

    Article  PubMed  Google Scholar 

  • Langa S, Maldonado A, Delgado S, Martín R, Martín V, Jiménez E, Ruíz-Barba JL, Mayo B, Connor RI, Suárez JE, Rodríguez JM (2012) Characterization of Lactobacillus salivarius CECT 5713, a strain isolated from human milk: from genotype to phenotype. Appl Microbiol Biotechnol 94:1279–1287

    Article  CAS  PubMed  Google Scholar 

  • Lara-Villoslada F, Sierra S, Díaz-Ropero MP, Rodríguez JM, Xaus J, Olivares M (2009) Safety assessment of Lactobacillus fermentum CECT5716, a probiotic strain isolated from human milk. J Dairy Res 76:216–221

    Article  CAS  PubMed  Google Scholar 

  • LeBlanc JG, Piard JC, Sesma F, de Giori GS (2005) Lactobacillus fermentum CRL 722 is able to deliver active alpha-galactosidase activity in the small intestine of rats. FEMS Microbiol Lett 248:177–182

    Article  CAS  PubMed  Google Scholar 

  • LeBlanc JG, Giori GSD, Smid EJ, Hugenholtz J, Sesma F (2007) Folate production by lactic acid bacteria and other food-grade microorganisms. Commun Curr Res Educ Topics Trends Appl Microbiol 1:329–339

    Google Scholar 

  • LeBlanc JG, Ledue-Clier F, Bensaada M, de Giori GS, Guerekobaya T, Sesma F, Juillard V, Rabot S, Piard JC (2008) Ability of Lactobacillus fermentum to overcome host alpha-galactosidase deficiency, as evidenced by reduction of hydrogen excretion in rats consuming soya alpha-galacto-oligosaccharides. BMC Microbiol 8:22

    Article  PubMed Central  PubMed  Google Scholar 

  • LeBlanc JG, Milani C, Savoy de Giori GS, Sesma F, van Sinderen D, Ventura M (2013) Bacteria as vitamin suppliers to their host: a gut microbiota perspective. Curr Opin Biotechnol 24:160–168

    Article  CAS  PubMed  Google Scholar 

  • Loguercio C, D’Argenio G, Delle Cave M, Cosenza V, Della Valle N, Mazzacca G, Del Vecchio BC (2003) Glutathione supplementation improves oxidative damage in experimental colitis. Dig Liver Dis 35:635–641

    Article  CAS  PubMed  Google Scholar 

  • Lu L, Hsieh M, Oriss TB, Morelf PA, Starzl TE, Rao AS, Thomson AW (1995) Generation of DC from mouse spleen cell cultures in response to GM-CSF: immunophenotypic and functional analyses. Immunology 84:127–134

    PubMed Central  CAS  PubMed  Google Scholar 

  • Maldonado J, Cañabate F, Sempere L, Vela F, Sánchez AR, Narbona E, López-Huertas E, Geerlings A, Valero AD, Olivares M, Lara-Villoslada F (2012) Human milk probiotic Lactobacillus fermentum CECT5716 reduces the incidence of gastrointestinal and upper respiratory tract infections in infants. J Pediatr Gastroenterol Nutr 54:55–61

    Article  PubMed  Google Scholar 

  • Mañé J, Lorén V, Pedrosa E, Ojanguren I, Xaus J, Cabré E, Domènech E, Gassull MA (2009) Lactobacillus fermentum CECT5716 prevents and reverts intestinal damage on TNBS-induced colitis in mice. Inflamm Bowel Dis 15:1155–1163

    Article  PubMed  Google Scholar 

  • Martín R, Langa S, Reviriego C, Jimenez E, Marin ML, Xaus J, Fernández L, Rodríguez JM (2003) Human milk is a source of lactic acid bacteria for the infant gut. J Pediatr 143:754–758

    Article  PubMed  Google Scholar 

  • Martin R, Olivares M, Marin ML, Fernandez L, Xaus J, Rodríguez JM (2005) Probiotic potential of 3 lactobacilli strains isolated from breast milk. J Hum Lact 21:8–17

    Article  PubMed  Google Scholar 

  • Martín R, Olivares M, Marín ML, Xaus J, Fernández L, Rodríguez JM (2005) Characterization of a reuterin-producing Lactobacillus coryniformis strain isolated from a goat’s milk cheese. Int J Food Microbiol 104:267–277

    Article  PubMed  Google Scholar 

  • Martín R, Olivares M, Pérez M, Xaus J, Torre C, Fernández L, Rodríguez JM (2010) Identification and evaluation of the probiotic potential of lactobacilli isolated from canine milk. Vet J 185:193–198

    Article  PubMed  Google Scholar 

  • Miralles-Barrachina O, Savoye G, Belmonte-Zalar L, Hochain P, Ducrotte P, Hecketsweiler B, Lerebours E, Dechelotte P (1999) Low levels of glutathione in endoscopic biopsies of patients with Crohn’s colitis: the role of malnutrition. Clin Nutr 18:313–317

    Article  CAS  PubMed  Google Scholar 

  • Morita H, Toh H, Fukuda S, Horikawa H, Oshima K, Suzuki T, Murakami M, Hisamatsu S, Kato Y, Takizawa T, Fukuoka H, Yoshimura T, Itoh K, O’Sullivan DJ, McKay LL, Ohno H, Kikuchi J, Masaoka T, Hattori M (2008) Comparative genome analysis of Lactobacillus reuteri and Lactobacillus fermentum reveal a genomic island for reuterin and cobalamin production. DNA Res 15:151–161

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Olivares M, Díaz-Ropero MP, Martín R, Rodríguez JM, Xaus J (2006) Antimicrobial potential of four Lactobacillus strains isolated from breast milk. J Appl Microbiol 101:72–79

    Article  CAS  PubMed  Google Scholar 

  • Olivares M, Díaz-Ropero MP, Sierra S, Lara-Villoslada F, Fonollá J, Navas M, Rodríguez JM, Xaus J (2007) Oral intake of Lactobacillus fermentum CECT5716 enhances the effects of influenza vaccination. Nutrition 23:254–260

    Article  CAS  PubMed  Google Scholar 

  • Peran L, Camuesco D, Comalada M, Nieto A, Concha A, Adrio JL, Olivares M, Xaus J, Zarzuelo A, Galvez J (2006) Lactobacillus fermentum, a probiotic capable to release glutathione, prevents colonic inflammation in the TNBS model of rat colitis. Int J Colorectal Dis 21:737–746

    Article  PubMed  Google Scholar 

  • Pérez-Cano FJ, Dong H, Yaqoob P (2010) In vitro immunomodulatory activity of Lactobacillus fermentum CECT5716 and Lactobacillus salivarius CECT5713: two probiotic strains isolated from human breast milk. Immunobiology 12:996–1004

    Article  Google Scholar 

  • Piddock LJ (2006) Multidrug-resistance efflux pumps—not just for resistance. Nat Rev Microbiol 4:629–636

    Article  CAS  PubMed  Google Scholar 

  • Santos F, Vera JL, van der Heijden R, Valdez G, de Vos WM, Sesma F, Hugenholtz J (2008) The complete coenzyme B12 biosynthesis gene cluster of Lactobacillus reuteri CRL1098. Microbiology 154:81–93

    Article  CAS  PubMed  Google Scholar 

  • Song Y-L, Kato N, Matsumiya Y, Liu C-X, Kato H, Watanabe K (1999) Identification of and hydrogen peroxide production by fecal and vaginal lactobacilli isolated from Japanese women and newborn infants. J Clin Microbiol 37:3062–3064

    PubMed Central  CAS  PubMed  Google Scholar 

  • Sybesma W, Starrenburg M, Tijsseling L, Hoefnagel MH, Hugenholtz J (2003) Effects of cultivation conditions on folate production by lactic acid bacteria. Appl Environ Microbiol 69:4542–4548

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Taranto MP, Vera JL, Hugenholtz J, De Valdez GF, Sesma F (2003) Lactobacillus reuteri CRL1098 produces cobalamin. J Bacteriol 185:5643–5647

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Yap PS, Gilliland SE (2000) Comparison of newly isolated strains of Lactobacillus delbrueckii susp. lactis for hydrogen peroxide production at 5°C. J Dairy Sci 83:628–632

    Article  CAS  PubMed  Google Scholar 

  • Zhang Z, Schwartz S, Wagner L, Miller W (2000) A greedy algorithm for aligning DNA sequences. J Comput Biol 7:203–214

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by CSD2007-00063 (FUN-C-FOOD, Consolider-Ingenio 2010), CYTED-IBEROFUN, and AGL2013-41980-P projects from the Ministerio de Ciencia e Innovación (Spain), a research contract funded by Biosearch Life (Granada, Spain) and by funding from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) and Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), Argentina.

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Authors and Affiliations

  1. Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Universidad Complutense de Madrid, Ciudad Universitaria, Avda. Puerta de Hierro, s/n., 28040, Madrid, Spain

    Nivia Cárdenas, Esther Jiménez, Leónides Fernández & Juan M. Rodríguez

  2. Probisearch, Tres Cantos, 28760, Madrid, Spain

    Nivia Cárdenas, Esther Jiménez, Leónides Fernández & Juan M. Rodríguez

  3. CERELA-CONICET, Centro de Referencia para Lactobacilos, Chacabuco 145, 4000, San Miguel de Tucumán, Argentina

    Jonathan E. Laiño, Marianela Juárez del Valle, Graciela Savoy de Giori, Fernando Sesma & Jean Guy LeBlanc

  4. Instituto de Productos Lácteos de Asturias (IPLA-CSIC), 33300, Villaviciosa, Spain

    Susana Delgado & Baltasar Mayo

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  1. Nivia Cárdenas
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  2. Jonathan E. Laiño
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  3. Susana Delgado
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  4. Esther Jiménez
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  5. Marianela Juárez del Valle
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  6. Graciela Savoy de Giori
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  7. Fernando Sesma
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  8. Baltasar Mayo
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  9. Leónides Fernández
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  10. Jean Guy LeBlanc
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  11. Juan M. Rodríguez
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Correspondence to Juan M. Rodríguez.

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Cárdenas, N., Laiño, J.E., Delgado, S. et al. Relationships between the genome and some phenotypical properties of Lactobacillus fermentum CECT 5716, a probiotic strain isolated from human milk. Appl Microbiol Biotechnol 99, 4343–4353 (2015). https://doi.org/10.1007/s00253-015-6429-0

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  • DOI: https://doi.org/10.1007/s00253-015-6429-0

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