Autochthonous lactic acid bacteria—presentation of potential probiotics application

  • Mirjana Ž. GrujovićEmail author
  • Katarina G. Mladenović
  • Danijela D. Nikodijević
  • Ljiljana R. Čomić
Original Research Paper



The objective of this study was to evaluate the probiotic potential as well as the ability of adhesion and aggregation of natural and autochthonous lactic acid bacteria, isolated from traditionally made cheese.


Lactic acid bacteria from natural food sources can be promising probiotic candidates and they can be used in natural food preservation or like starter cultures. Tested autochthonous isolates showed tolerance to the simulated gastrointestinal condition as well as the sensitivity to clinically relevant antibiotics, especially to ampicillin (MIC at 0.195 μg mL−1 for lactobacilli and from 0.195 to 3.125 μg mL−1 for lactococci). Among isolates, the highest percentage of adhesion was detected with chloroform, while the adhesion ability of selected isolates to pig intestinal epithelium was in the correlation with the results of adhesion ability with solvents. The auto-aggregation ability of isolates was demonstrated, while co-aggregation with Escherichia coli was strain specific.


The results indicated the potential probiotic properties of the isolates and give evidence for further investigation and potential application in the dairy industry.


Antibiotic Adhesion Aggregation ability Lactic acid bacteria Probiotics Pig intestinal epithelium 



This investigation was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (Grant No. 41010).

Author contributions

MŽG conceived and designed the experiments; DDN designed and photographed samples by florescent microscope and interpreted the results of adhesion; MŽG and KGM processed the results; KGM and LRČ take charge of the preparation of the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest with the current work or its publication.

Research involving human and animal participants

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. Bassyouni RH, Abdel-all WS, Fadl MG, Abdel-all S, Kamel Z (2012) Characterization of lactic acid bacteria isolated from dairy products in Egypt as a probiotic. Life Sci J 9:2924–2933Google Scholar
  2. Carasi P, Ambrosis NM, De Antoni GL, Bressollier P, Urdaci MC, de los Angeles Serradell M (2014) Adhesion properties of potentially probiotic Lactobacillus kefiri to gastrointestinal mucus. J Dairy Res 81(1):16–23CrossRefGoogle Scholar
  3. Collado MC, Meriluoto J, Salminen S (2008) Adhesion and aggregation properties of probiotic and pathogen strains. Eur Food Res Technol 226:1065–1073CrossRefGoogle Scholar
  4. Dowarah R, Verma AK, Agarwal N, Singh P, Singh BR (2018) Selection and characterization of probiotic lactic acid bacteria and its impact on growth, nutrient digestibility, health and antioxidant status in weaned piglets. PLoS ONE 13(3):e0192978. CrossRefGoogle Scholar
  5. Elhadidy M, Zahran E (2014) Biofilm mediates Enterococcus faecalis adhesion, invasion and survival into bovine mammary epithelial cells. Lett Appl Microbiol 58:248–254CrossRefGoogle Scholar
  6. European Food Safety Authority-EFSA (2012) Guidance on the assessment of bacterial susceptibility to antimicrobials of human or veterinary importance. EFSA J 10:1–10Google Scholar
  7. FAO/WHO (World Health Organization) (2006) Probiotics in food. Health and nutritional properties and guidelines for evaluation. FAO Food Nutr Pap 85:2.Google Scholar
  8. Garriga M, Rubio R, Aymerich T, Ruas-Madiedo P (2014) Potentially probiotic and bioprotective lactic acid bacteria starter cultures antagonise the Listeria monocytogenes adhesion to HT29 colonocyte-like cells. Benef Microbes 6(3):337–343CrossRefGoogle Scholar
  9. Hernandez-Hernandez O, Muthaiyan A, Moreno FJ, Montilla A, Sanz ML, Rickeet SC (2012) Effect of prebiotic carbohydrates on the growth and tolerance of Lactobacillus. Food Microbiol 30:355–361CrossRefGoogle Scholar
  10. Huang Y, Adams MC (2004) In vitro assessment of the upper gastrointestinal tolerance of potential probiotic dairy propionibacteria. Int J Food Microbiol 91:253–260CrossRefGoogle Scholar
  11. Idou T (2014) Probiotic properties of Lactobacillus strains isolated from gizzard of local poultry. Iran J Microbiol 6(2):120–126Google Scholar
  12. Janković T, Frece J, Abram M, Gobin I (2012) Aggregation ability of potential probiotic Lactobacillus plantarum strains. Int J Sanit Eng Res 6:19–24Google Scholar
  13. Jeong DW, Lee JH (2015) Antibiotic resistance, hemolysis and biogenic amine production assessments of Leuconostoc and Weissella isolates for kimchi starter development. LWT Food Sci Technol 64:1078–1084CrossRefGoogle Scholar
  14. Kaewnopparat S, Dangmanee N, Kaewnopparat N, Srichana T, Chulasiri M, Settharaksa S (2013) In vitro probiotic properties of Lactobacillus fermentum SK5 isolated from vagina of a healthy woman. Anaerobe 22:6–13CrossRefGoogle Scholar
  15. Kaktcham PM, Zambou NF, Tchouanguep FM, El-Soda M, Choudhary MI (2012) Antimicrobial and safety properties of lactobacilli isolated from two Cameroonian traditional fermented foods. Sci Pharm 80:189–203CrossRefGoogle Scholar
  16. Kos B, Šušković J, Vuković S, Šimpraga M, Frece J, Matošić S (2003) Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J Appl Microbiol 94:981–987CrossRefGoogle Scholar
  17. Kumar AM, Murugalatha N (2012) Isolation of Lactobacillus plantarum from cow milk and screening for the presence of sugar alcohol producing gene. J Clin Microbiol Antimicrob 4:16–22Google Scholar
  18. Lee YK, Salminen S (2009) Handbook of probiotics and prebiotics. Wiley, pp 386Google Scholar
  19. Leite AMO, Miguel MAL, Peixoto RS, Ruas-Madiedo P, Paschoalin VMF, Mayo B, Delgado S (2015) Probiotic potential of selected lactic acid bacteria strains isolated from Brazilian kefir grains. J Dairy Sci 98:3622–3632CrossRefGoogle Scholar
  20. Li Q, Liu X, Dong M, Zhou J, Wang Y (2015) Aggregation and adhesion abilities of 18 lactic acid bacteria strains isolated from traditional fermented food. Int J Agric Policy Res 3(2):84–92Google Scholar
  21. Magdoub MNI, Hassan ZMR, Effat BAM, Sadek ZIM, Tawfik NF, Mabrouk AMM (2015) Probiotic properties of some lactic acid bacteria isolated from Egyptian dairy products. Int J Curr Microbiol Appl Sci 4:758–766Google Scholar
  22. Muruzović MŽ, Mladenović KG, Stefanović OD, Vasić SM, Čomić LR (2016) Extracts of Agrimonia eupatoria L. as sources of biologically active compounds and evaluation of their antioxidant, antimicrobial, and antibiofilm activities. J Food Drug Anal 24:539–547CrossRefGoogle Scholar
  23. Muruzović MŽ, Mladenović KG, Žugić Petrović TD, Čomić LR (2018a) Characterization of lactic acid bacteria isolated from traditionally made Serbian cheese and evaluation of their antagonistic potential against Enterobacteriaceae. J Food Process Preserv 42(4):e13577. CrossRefGoogle Scholar
  24. Muruzović MŽ, Mladenović KG, Đilas MD, Stefanović OD, Čomić LR (2018b) In vitro evaluation of antimicrobial potential and ability of biofilm formation of autochthonous Lactobacillus spp. and Lactococcus spp. isolated from traditionally made cheese from Southeastern Serbia. J Food Process Preserv 42(11):13776Google Scholar
  25. Muruzović MŽ, Mladenović KG, Čomić LR (2018) In vitro evaluation of resistance to environmental stress by planktonic and biofilm form of lactic acid bacteria isolated from traditionally made cheese from Serbia. Food Biosci 23:54–59CrossRefGoogle Scholar
  26. Ocaña V, Nader-Macías ME (2002) Vaginal lactobacilli: self- and co-aggregation ability. Br J Biomed Sci 59:183–190CrossRefGoogle Scholar
  27. Palomares IC, Morales PR, Felix AE (2007) Evaluation of probiotic properties in Lactobacillus isolated from small intestine of piglets. Rev Latinoameric Microbiol 49(3–4):46–54Google Scholar
  28. Pavli PG, Argyri AA, Papadopoulou OS, Nychas GE, Chorianopoulos NG, Tassou CC (2016) Probiotic potential of lactic acid bacteria from traditional fermented dairy and meat products: assessment by in vitro tests and molecular characterization. J Probiotics Health 4:157–165CrossRefGoogle Scholar
  29. Pereira NG, Figueiredo FJB, Dias-Souza MV (2015) Antimicrobial susceptibility of commercial probiotic Lactobacillus strains. J Appl Pharm Sci 2(2):14–17Google Scholar
  30. Popović N, Dinić M, Tolinački M, Mihajlović S, Terzić-Vidojević A, Bojić S, Djokić J, Golić N, Veljović K (2018) New insight into biofilm formation ability, the presence of virulence genes and probiotic potential of Enterococcus sp. dairy isolates. Front Microbiol 9:78CrossRefGoogle Scholar
  31. Pringsulaka O, Rueangyotchanthana K, Suwannasai N, Watanapokasin N, Amnueysit P, Sunthornthummas S, Sukkhum A, Sarawaneeyaruk S, Rangsiruji A (2015) In vitro screening of lactic acid bacteria for multi-strain probiotics. Livestock Sci 174:66–73CrossRefGoogle Scholar
  32. Ramos CL, Thorsen L, Schwan RF, Jespersen L (2013) Strain-specific probiotics properties of Lactobacillus fermentum, Lactobacillus plantarum and Lactobacillus brevis isolates from Brazilian food products. Food Microbiol 36:22–29CrossRefGoogle Scholar
  33. Rosenberg M, Gutnick D, Rosenberg E (1980) Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Lett 9:29–33CrossRefGoogle Scholar
  34. Sarker SD, Nahar L, Kumarasamy Y (2007) Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods 42:321–324CrossRefGoogle Scholar
  35. Shokryazdan P, Sieo CC, Kalavathy R, Liang JB, Alithee NB, Jahromi MF, Ho YW (2014) Probiotic potential of Lactobacillus strains with antimicrobial activity against some human pathogenic strains. BioMed Res Int 2014:1–16CrossRefGoogle Scholar
  36. Sitepu GR, Nursyirwani N, Efriyeldi E (2016) Adhesion of lactic acid bacteria (LAB) to intestinal epithelial cells of red snapper (Lutjanus argentimaculatus) in inhibiting Vibrio alginolyticus. J Online Mahasiswa 3(2):1–10Google Scholar
  37. Solieri L, Bianchi A, Mottolese G, Lemmetti F, Giudici P (2014) Tailoring the probiotic potential of non-starter Lactobacillus strains from ripened Parmigiano Reggiano cheese by in vitro screening and principal component analysis. Food Microbiol 38:240–249CrossRefGoogle Scholar
  38. Šušković J, Kos B, Goreta J, Matosić S (2001) Role of lactic acid bacteria and bifidobacteria in symbiotic effect. Food Technol Biotechnol 39:227–235Google Scholar
  39. Tuo Y, Yu H, Ai L, Wu Z, Guo B, Chen W (2013) Aggregation and adhesion properties of 22 Lactobacillus strains. J Dairy Sci 96:4252–4257CrossRefGoogle Scholar
  40. Uroić K, Nikolić M, Koslć B, Pavunc L, Beganović J, Lukić J, Jovčić B, Filipić B, Miljković M, Golić N, Topisirović L, Čadež N, Raspor P, Šušković J (2014) Probiotic properties of lactic acid bacteria isolated from Croatian fresh soft cheese and Serbian white pickled cheese. Food Technol Biotechnol 52:232–241Google Scholar
  41. Vesković Moračanin S, Djukić D, Zdolec N, Milijašević M, Mašković P (2017) Antimicrobial resistance of lactic acid bacteria in fermented food. J Hyg Eng Des 18:25–35Google Scholar
  42. Walsham ADS, MacKenzie DA, Cook V, Wemyss-Holden S, Hews CL, Juge N, Schüller S (2016) Lactobacillus reuteri inhibition of enteropathogenic Escherichia coli adherence to human intestinal epithelium. Front Microbiol 7:244CrossRefGoogle Scholar
  43. Younes JA, van der Mei HC, van den Heuvel E, Busscher HJ, Reid G (2012) Adhesion forces and coaggregation between vaginal staphylococci and lactobacilli. PLoS ONE 7(5):1–8CrossRefGoogle Scholar
  44. Živković M, Miljković MS, Ruas-Madiedo P, Markelić MB, Veljović K, Tolinački M, Sokocić S, Korać A, Golić N (2016) EPS-SJ exopolisaccharide produced by the strain Lactobacillus paracasei subsp. paracasei BGSJ2-8 is involved in adhesion to epithelial intestinal cells and decrease on E. coli association to Caco-2 cells. Front Microbiol 7:286Google Scholar
  45. Zoumpopoulou G, Foligne B, Christodoulou K, Grangette C, Pot B, Tsakalidou E (2008) Lactobacillus fermentum ACA-DC 179 displays probiotic potential in vitro and protects against trinitrobenzene sulfonic acid (TNBS)-induced colitis and Salmonella infection in murine models. Int J Food Microbiol 121:18–26CrossRefGoogle Scholar

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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Biology and Ecology, Faculty of ScienceUniversity of KragujevacKragujevacRepublic of Serbia

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