Putative probiotic lactic acid bacteria isolated from sauerkraut fermentations

Tiago Touret, Manuela Oliveira, Teresa Semedo-Lemsaddek, Tiago Touret, Manuela Oliveira, Teresa Semedo-Lemsaddek

Abstract

Probiotics are live microorganisms which confer health benefits to the host, and may be isolated from several sources, such as vegetable foodstuffs. Sauerkraut is a cabbage product resulting from fermentation by a lactic acid bacteria microbial succession, and is a potential source for probiotics. The aim of the present study was the isolation and characterization of probiotic microorganisms from sauerkraut fermentations. Four distinct fermentations were performed, from which lactic acid bacteria were recovered. Overall, 114 isolates were obtained, phenotypically and genotypically characterized, identified to the genus level and evaluated regarding safety and probiotic potential. Representative bacteria were selected for further analysis, 52% being Lactobacillus spp. and 33% belonging to Leuconostoc spp. genus. One isolate revealed to be β-hemolytic, 42% possessed potentially mobile antimicrobial resistance, 88% were resistant to bile and 20% to low pH. The six most promising candidates were further characterized and presented antimicrobial activity against Listeria monocytogenes, three being resistant to lower pH values. Thus, global analysis of data gathered during this study highlighted the identification of three Lactobacillus strains with putative probiotic potential, suggesting the applicability of sauerkraut fermentations as a source for probiotic isolation. Due to their origin these strains should be suited for future application in the food industry, namely vegetable products such as sauerkraut itself.

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1. Variation of pH (A), acid…
Fig 1. Variation of pH (A), acid content (B) and viable lactic acid bacteria (C) along fermentation time for the four distinct fermentations.
Fig 2. Dendrogram built using M13 and…
Fig 2. Dendrogram built using M13 and OPC-15 PCR-fingerprinting profiles of the 95 isolates from the four sauerkraut fermentations.
The vertical line represents the reproducibility level, which was used as a cut-off value for the definition of genomically distinct LAB. Isolates highlighted with a dot (●) were chosen as representatives for further studies.
Fig 3. Incidence of Leuconostoc and Lactobacillus…
Fig 3. Incidence of Leuconostoc and Lactobacillus isolates among representatives from the four sauerkraut fermentations.
Fisher’s exact test was used to determine statistically significant differences between fermentations. *—P

Fig 4. Dendrograms built using the PCR-fingerprinting…

Fig 4. Dendrograms built using the PCR-fingerprinting profiles of Lactobacillus (A) or Leuconostoc (B) isolates.

Fig 4. Dendrograms built using the PCR-fingerprinting profiles of Lactobacillus (A) or Leuconostoc (B) isolates.
The vertical line represents the reproducibility level, which was used as a cut-off value for the definition of genomically similar groups. Groups containing more than one isolate are represented. Isolates indicated with an arrow were considered genomically similar to others within the same fermentation/time-point, and were removed from subsequent characterization.

Fig 5. Percentage of isolates resistant to…

Fig 5. Percentage of isolates resistant to the studied antimicrobial compounds.

AMP- Ampicillin; C- Chloramphenicol;…

Fig 5. Percentage of isolates resistant to the studied antimicrobial compounds.
AMP- Ampicillin; C- Chloramphenicol; DA- Clindamycin; E- Erythromycin; CN- Gentamicin; K- Kanamycin; S- Streptomycin; TE- Tetracycline. Fisher’s exact test was used to determine statistically significant differences between genera. *—P

Fig 6. Summary of results for the…

Fig 6. Summary of results for the 95 isolates with a LAB-like phenotype.

Dendrogram built…

Fig 6. Summary of results for the 95 isolates with a LAB-like phenotype.
Dendrogram built based on PCR-fingerprinting profiles, with the red vertical line representing the reproducibility level. Information regarding hemolytic activity, antimicrobial resistance, low pH (3.5) and bile resistance, genus identification, group attributed after PCR-fingerprinting analysis and source of the isolate is also shown. Isolates written in red were chosen as representatives after PCR-fingerprinting, and isolates written in blue were also chosen, but excluded after further analysis. Isolates marked with a box were selected for subsequent analysis based on the information presented in the figure. Black squares represent the presence of AMP—Ampicillin; C—Chloramphenicol; DA—Clindamycin; E—Erythromycin; CN—Gentamicin; K—Kanamycin; S—Streptomycin; or TE—Tetracycline resistance.
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References
    1. FAO/WHO. Probiotics in food—Health and nutritional properties and guidelines for evaluation Food Nutr Pap. 2001;85.
    1. Fontana L, Bermudez-Brito M, Plaza-Diaz J, Muñoz-Quezada S, Gil A. Sources, isolation, characterisation and evaluation of probiotics. Br J Nutr. 2013;109 Suppl:S35–50. 10.1017/S0007114512004011 - DOI - PubMed
    1. Yadav R. Probiotics for Human Health: Current Progress and Applications In: Shukla P, editor. Recent advances in Applied Microbiology. Springer; 2017. p. 133–47. 10.1007/978-981-10-5275-0 - DOI
    1. Aureli P, Capurso L, Castellazzi AM, Clerici M, Giovannini M, Morelli L, et al. Probiotics and health: An evidence-based review. Pharmacol Res. 2011;63(5):366–76. 10.1016/j.phrs.2011.02.006 - DOI - PubMed
    1. Papadimitriou K, Zoumpopoulou G, Foligné B, Alexandraki V, Kazou M, Pot B, et al. Discovering probiotic microorganisms: in vitro, in vivo, genetic and omics approaches. Front Microbiol. 2015;6(58). 10.3389/fmicb.2015.00058 - DOI - PMC - PubMed
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The authors would like to thank CIISA – Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon for the financial support (Project UID/CVT/276/2013). T. Semedo-Lemsaddek holds a scholarship from “Fundação para a Ciência e Tecnologia” (SFRH/BPD/108123/2015).
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Fig 4. Dendrograms built using the PCR-fingerprinting…
Fig 4. Dendrograms built using the PCR-fingerprinting profiles of Lactobacillus (A) or Leuconostoc (B) isolates.
The vertical line represents the reproducibility level, which was used as a cut-off value for the definition of genomically similar groups. Groups containing more than one isolate are represented. Isolates indicated with an arrow were considered genomically similar to others within the same fermentation/time-point, and were removed from subsequent characterization.
Fig 5. Percentage of isolates resistant to…
Fig 5. Percentage of isolates resistant to the studied antimicrobial compounds.
AMP- Ampicillin; C- Chloramphenicol; DA- Clindamycin; E- Erythromycin; CN- Gentamicin; K- Kanamycin; S- Streptomycin; TE- Tetracycline. Fisher’s exact test was used to determine statistically significant differences between genera. *—P

Fig 6. Summary of results for the…

Fig 6. Summary of results for the 95 isolates with a LAB-like phenotype.

Dendrogram built…

Fig 6. Summary of results for the 95 isolates with a LAB-like phenotype.
Dendrogram built based on PCR-fingerprinting profiles, with the red vertical line representing the reproducibility level. Information regarding hemolytic activity, antimicrobial resistance, low pH (3.5) and bile resistance, genus identification, group attributed after PCR-fingerprinting analysis and source of the isolate is also shown. Isolates written in red were chosen as representatives after PCR-fingerprinting, and isolates written in blue were also chosen, but excluded after further analysis. Isolates marked with a box were selected for subsequent analysis based on the information presented in the figure. Black squares represent the presence of AMP—Ampicillin; C—Chloramphenicol; DA—Clindamycin; E—Erythromycin; CN—Gentamicin; K—Kanamycin; S—Streptomycin; or TE—Tetracycline resistance.
Fig 6. Summary of results for the…
Fig 6. Summary of results for the 95 isolates with a LAB-like phenotype.
Dendrogram built based on PCR-fingerprinting profiles, with the red vertical line representing the reproducibility level. Information regarding hemolytic activity, antimicrobial resistance, low pH (3.5) and bile resistance, genus identification, group attributed after PCR-fingerprinting analysis and source of the isolate is also shown. Isolates written in red were chosen as representatives after PCR-fingerprinting, and isolates written in blue were also chosen, but excluded after further analysis. Isolates marked with a box were selected for subsequent analysis based on the information presented in the figure. Black squares represent the presence of AMP—Ampicillin; C—Chloramphenicol; DA—Clindamycin; E—Erythromycin; CN—Gentamicin; K—Kanamycin; S—Streptomycin; or TE—Tetracycline resistance.

References

    1. FAO/WHO. Probiotics in food—Health and nutritional properties and guidelines for evaluation Food Nutr Pap. 2001;85.
    1. Fontana L, Bermudez-Brito M, Plaza-Diaz J, Muñoz-Quezada S, Gil A. Sources, isolation, characterisation and evaluation of probiotics. Br J Nutr. 2013;109 Suppl:S35–50. 10.1017/S0007114512004011
    1. Yadav R. Probiotics for Human Health: Current Progress and Applications In: Shukla P, editor. Recent advances in Applied Microbiology. Springer; 2017. p. 133–47. 10.1007/978-981-10-5275-0
    1. Aureli P, Capurso L, Castellazzi AM, Clerici M, Giovannini M, Morelli L, et al. Probiotics and health: An evidence-based review. Pharmacol Res. 2011;63(5):366–76. 10.1016/j.phrs.2011.02.006
    1. Papadimitriou K, Zoumpopoulou G, Foligné B, Alexandraki V, Kazou M, Pot B, et al. Discovering probiotic microorganisms: in vitro, in vivo, genetic and omics approaches. Front Microbiol. 2015;6(58). 10.3389/fmicb.2015.00058
    1. Grazul H, Kanda LL, Gondek D. Impact of probiotic supplements on microbiome diversity following antibiotic treatment of mice. Gut Microbes. 2016;7(2):101–14. 10.1080/19490976.2016.1138197
    1. Gareau MG, Sherman PM, Walker WA. Probiotics and the gut microbiota in intestinal health and disease. Nat Rev Gastroenterol Hepatol. 2010;7(9):503–14. 10.1038/nrgastro.2010.117
    1. Dahiya DK, Renuka, Puniya M, Shandilya UK, Dhewa T, Kumar N, et al. Gut microbiota modulation and its relationship with obesity using prebiotic fibers and probiotics: A review. Front Microbiol. 2017;8(563). 10.3389/fmicb.2017.00563
    1. Yadav R, Shukla P. An overview of advanced technologies for selection of probiotics and their expediency: A review. Crit Rev Food Sci Nutr. 2017;57(15):3233–42. 10.1080/10408398.2015.1108957
    1. Peres CM, Peres C, Hernández-Mendoza A, Malcata FX. Review on fermented plant materials as carriers and sources of potentially probiotic lactic acid bacteria—With an emphasis on table olives. Trends Food Sci Technol. 2012;26(1):31–42.
    1. Harris LJ. The microbiology of vegetable fermentations In: Wood BJB, editor. Microbiology of Fermented Foods. Boston, MA: Springer US; 1998. p. 45–72.
    1. Holzapfel W, Schillinger U, Buckenhüskes H. Sauerkraut In: Farnworth ER, editor. Handbook of Fermented Functional Foods. 2nd ed CRC Press; 2008. p. 395–412.
    1. Zabat MA, Sano WH, Wurster JI, Cabral DJ, Belenky P. Microbial Community Analysis of Sauerkraut Fermentation Reveals a Stable and Rapidly Established Community. Foods. 2018;7(77). 10.3390/foods7050077
    1. Plengvidhya V, Breidt F, Lu Z, Fleming HP. DNA fingerprinting of lactic acid bacteria in sauerkraut fermentations. Appl Environ Microbiol. 2007;73(23):7697–702. 10.1128/AEM.01342-07
    1. Beganović J, Kos B, Leboš Pavunc A, Uroić K, Jokić M, Šušković J. Traditionally produced sauerkraut as source of autochthonous functional starter cultures. Microbiol Res. 2014;169(7–8):623–32. 10.1016/j.micres.2013.09.015
    1. Chang JH, Shim YY, Cha SK, Chee KM. Probiotic characteristics of lactic acid bacteria isolated from kimchi. J Appl Microbiol. 2010;109(1):220–30. 10.1111/j.1365-2672.2009.04648.x
    1. Yu Z, Zhang X, Li S, Li C, Li D, Yang Z. Evaluation of probiotic properties of Lactobacillus plantarum strains isolated from Chinese sauerkraut. World J Microbiol Biotechnol. 2013;29(3):489–98. 10.1007/s11274-012-1202-3
    1. Millar BC, Jiru X, Moore JE, Earle JA. A simple and sensitive method to extract bacterial, yeast and fungal DNA from blood culture material. J Microbiol Methods. 2000;42(2):139–47. 10.1016/S0167-7012(00)00174-3
    1. Dubernet S, Desmasures N, Guéguen M. A PCR-based method for identification of lactobacilli at the genus level. FEMS Microbiol Lett. 2002;214(2):271–5. 10.1016/S0378-1097(02)00895-9
    1. Macián MC, Chenoll E, Aznar R. Simultaneous detection of Carnobacterium and Leuconostoc in meat products by multiplex PCR. J Appl Microbiol. 2004;97(2):384–94. 10.1111/j.1365-2672.2004.02317.x
    1. CLSI. Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved Standard M02-A11—Eleventh Edition. Clinical and Laboratory Standards Institute; 2012.
    1. Argyri AA, Zoumpopoulou G, Karatzas KA, Tsakalidou E, Nychas GJ, Panagou EZ, et al. Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests. Food Microbiol. 2013;33(2):282–91. 10.1016/j.fm.2012.10.005
    1. Peres CM, Alves M, Hernandez-Mendoza A, Moreira L, Silva S, Bronze MR, et al. Novel isolates of lactobacilli from fermented Portuguese olive as potential probiotics. LWT—Food Sci Technol. 2014;59(1):234–46. 10.1016/j.lwt.2014.03.003
    1. Botta C, Langerholc T, Cencič A, Cocolin L. In vitro selection and characterization of new probiotic candidates from table olive microbiota. PLoS One. 2014;9(4):e94457 10.1371/journal.pone.0094457
    1. Lu Z, Breidt F, Plengvidhya V, Fleming HP. Bacteriophage ecology in commercial sauerkraut fermentations. Appl Environ Microbiol. 2003;69(6):3192–202. 10.1128/AEM.69.6.3192-3202.2003
    1. Rosa E, David M, Gomes MH. Glucose, fructose and sucrose content in broccoli, white cabbage and Portuguese cabbage grown in early and late seasons. J Sci Food Agric. 2001;81(12):1145–9. 10.1002/jsfa.919
    1. Lee J, Yun HS, Cho KW, Oh S, Kim SH, Chun T, et al. Evaluation of probiotic characteristics of newly isolated Lactobacillus spp.: immune modulation and longevity. Int J Food Microbiol. 2011;148(2):80–6. 10.1016/j.ijfoodmicro.2011.05.003
    1. Bernardeau M, Vernoux JP, Henri-Dubernet S, Guéguen M. Safety assessment of dairy microorganisms: The Lactobacillus genus. Int J Food Microbiol. 2008;126(3):278–85. 10.1016/j.ijfoodmicro.2007.08.015
    1. Ammor MS, Flórez AB, Mayo B. Antibiotic resistance in non-enterococcal lactic acid bacteria and bifidobacteria. Food Microbiol. 2007;24(6):559–70. 10.1016/j.fm.2006.11.001
    1. Ogier JC, Casalta E, Farrokh C, Saïhi A. Safety assessment of dairy microorganisms: the Leuconostoc genus. Int J Food Microbiol. 2008;126(3):286–90. 10.1016/j.ijfoodmicro.2007.08.012
    1. Devirgiliis C, Zinno P, Perozzi G. Update on antibiotic resistance in foodborne Lactobacillus and Lactococcus species. Front Microbiol. 2013;4:301 10.3389/fmicb.2013.00301
    1. Fraqueza MJ. Antibiotic resistance of lactic acid bacteria isolated from dry-fermented sausages. Int J Food Microbiol. 2015;212:76–88. 10.1016/j.ijfoodmicro.2015.04.035
    1. Kastner S, Perreten V, Bleuler H, Hugenschmidt G, Lacroix C, Meile L. Antibiotic susceptibility patterns and resistance genes of starter cultures and probiotic bacteria used in food. Syst Appl Microbiol. 2006;29(2):145–55. 10.1016/j.syapm.2005.07.009
    1. Gahan CG, Hill C. Listeria monocytogenes: survival and adaptation in the gastrointestinal tract. Front Cell Infect Microbiol. 2014;4:9 10.3389/fcimb.2014.00009
    1. Cotter PD, Hill C, Ross RP. Bacteriocins: developing innate immunity for food. Nat Rev Microbiol. 2005;3(10):777–88. 10.1038/nrmicro1273
    1. Son SH, Jeon HL, Jeon EB, Lee NK, Park YS, Kang DK, et al. Potential probiotic Lactobacillus plantarum Ln4 from kimchi: Evaluation of β-galactosidase and antioxidant activities. LWT—Food Sci Technol. 2017;85:181–6. 10.1016/j.lwt.2017.07.018
    1. Lee H, Yoon H, Ji Y, Kim H, Park H, Lee J, et al. Functional properties of Lactobacillus strains isolated from kimchi. Int J Food Microbiol. 2011;145(1):155–61. 10.1016/j.ijfoodmicro.2010.12.003
    1. Min M, Bunt CR, Mason SL, Hussain MA. Non-dairy probiotic food products: An emerging group of functional foods. Crit Rev Food Sci Nutr. 2018;1–16. 10.1080/10408398.2018.1462760

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