Advertisement

Annals of Microbiology

, Volume 69, Issue 12, pp 1275–1287 | Cite as

Isolation, technological characterization and in vitro probiotic evaluation of Lactococcus strains from traditional Turkish skin bag Tulum cheeses

  • Erhan Kazancıgil
  • Talha Demirci
  • Hale İnci Öztürk-NegişEmail author
  • Nihat Akın
Original Article
  • 178 Downloads

Abstract

Purpose

The present study was undertaken to evaluate in vitro prerequisite probiotic and technological characteristics of ten Lactococcus strains isolated from traditional goat skin bags of Tulum cheeses from the Central Taurus mountain range in Turkey.

Methods

All isolates were identified based on the nucleotide sequences of the 16S rRNA gene. Eight isolates belonged to Lactococcus lactis and two belonged to Lactococcus garvieae. Probiotic potential was determined from resistance to acid and bile salt, resistance to gastric and pancreatic juices, resistance to antibiotic, auto-aggregation, co-aggregation, diacetyl, hydrogen peroxide and exopolysaccharide productions. Technological properties were verified by alcohol, NaCl and hydrogen peroxide resistance and temperature tests.

Results

L. lactis NTH7 displayed high growth at all alcohol concentrations while L. lactis NTH4 grew very well even at NaCl concentrations of 10%. All strains showed to some extent resistance to acid and bile. Five strains exhibited desirable survival in gastric juice (pH 2.0), while three strains survived in pancreatic juice (pH 8.0). All Lactococcus isolates were sensitive to ampicillin, chloramphenicol, erythromycin, vancomycin, kanamycin, gentamycin and tetracycline. Also, only L. lactis NTH7 from among the isolates showed resistance against penicillin. L. lactis NTH10 and L. lactis NTH7 had higher auto-aggregation values in comparison with all other strains. All the strains demonstrated a co-aggregation ability against model food pathogens, particularly, L. lactis NTH10 which showed a superior ability with L. monocytogenes. All the ten strains produced H2O2 and exopolysaccharide (EPS); however, diacetyl production was detected for only four strains including L. lactis NTH10.

Conclusion

These results demonstrate that the L. lactis NTH10 isolate could be regarded as a favorable probiotic candidate for future in vivo studies.

Keywords

Probiotic Technological characteristics Lactococcus Tulum cheese 

Notes

Funding information

TUBITAK (Scientific and Technological Research Council of Turkey) supported the isolation and identification of Lactococcus strains from traditional Turkish skin bag Tulum cheeses by the project number TOAG-214Z054.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

Not applicable.

Informed consent

Informed consent was obtained from all individual participants included in the study.

References

  1. Abushelaibi A, Al-Mahadin S, El-Tarabily K, Shah NP, Ayyash M (2017) Characterization of potential probiotic lactic acid bacteria isolated from camel milk. LWT-Food Sci Technol 79:316–325Google Scholar
  2. Adamberg K, Kask S, Laht T-M, Paalme T (2003) The effect of temperature and pH on the growth of lactic acid bacteria: a pH-auxostat study. Int J Food Microbiol 85:171–183PubMedGoogle Scholar
  3. An YH, Dickinson RB, Doyle RJ (2000) Mechanisms of bacterial adhesion and pathogenesis of implant and tissue infections. Handbook of Bacterial Adhesion. Springer, In, pp 1–27Google Scholar
  4. Attar MA, Yavarmanesh M, Mortazavi A, Dovom MRE, Najafi MBH (2018) Antibacterial effects of Lactococcus lactis isolated from Lighvan cheese regarding the recognition of Nisin, Lacticin and Lactococcin structural genes. LWT-Food Sci Technol 89:186–191Google Scholar
  5. Badis A, Guetarni D, Boudjema BM, Henni D, Kihal M (2004) Identification and technological properties of lactic acid bacteria isolated from raw goat milk of four Algerian races. Food Microbiol 21:579–588Google Scholar
  6. Bao Y et al (2010) Screening of potential probiotic properties of Lactobacillus fermentum isolated from traditional dairy products. Food Control 21:695–701Google Scholar
  7. Bautista-Gallego J, Arroyo-López F, Rantsiou K, Jiménez-Díaz R, Garrido-Fernández A, Cocolin L (2013) Screening of lactic acid bacteria isolated from fermented table olives with probiotic potential. Food Res Int 50:135–142Google Scholar
  8. Bengoa AA, Llamas MG, Iraporda C, Dueñas MT, Abraham AG, Garrote GL (2018) Impact of growth temperature on exopolysaccharide production and probiotic properties of Lactobacillus paracasei strains isolated from kefir grains. Food Microbiol 69:212–218PubMedGoogle Scholar
  9. Cakmakci S, Dagdemir E, Hayaloglu A, Gurses M, Gundogdu E (2008) Influence of ripening container on the lactic acid bacteria population in Tulum cheese. World J Microbiol Biotechnol 24:293–299Google Scholar
  10. Chalas R, Janczarek M, Bachanek T, Mazur E, Cieszko-Buk M, Szymanska J (2016) Characteristics of oral probiotics–a review. Curr Issues Pharm Med Sci 29:8–10Google Scholar
  11. Charteris WP, Kelly MP, Morelli L, Collins KJ (1998a) Antibiotic susceptibility of potentially probiotic Lactobacillus species. Journal Food Prot 61(12):1636–1164Google Scholar
  12. Charteris WP, Kelly MP, Morelli L, Collins KJ (1998b) Development and application of an in vitro methodology to determine the transit tolerance of potentially probiotic Lactobacillus and Bifidobacterium species in the upper human gastrointestinal tract. J Appl Microbiol 84:759–768PubMedGoogle Scholar
  13. Cogan TM et al (1997) Characterization of the lactic acid bacteria in artisanal dairy products. J Dairy Res 64:409–421Google Scholar
  14. Collins MD, Gibson GR (1999) Probiotics, prebiotics, and synbiotics: approaches for modulating the microbial ecology of the gut. Am J Clin Nutr 69:1052s-1057sPubMedGoogle Scholar
  15. Danielsen M, Wind A (2003) Susceptibility of Lactobacillus spp. to antimicrobial agents. Int J Food Microbiol 82(1):1–11PubMedGoogle Scholar
  16. Das P, Khowala S, Biswas S (2016) In vitro probiotic characterization of Lactobacillus casei isolated from marine samples. LWT-Food Sci Technol 73:383–390Google Scholar
  17. de Almeida Júnior WLG, da Silva FÍ, de Souza JV, da Silva CDA, da Costa MM, Dias FS (2015) Characterization and evaluation of lactic acid bacteria isolated from goat milk. Food Control 53:96–103Google Scholar
  18. DeLisle S, Perl TM (2003) Vancomycin-resistant enterococci: a road map on how to prevent the emergence and transmission of antimicrobial resistance. Chest 123(5):504S–518SPubMedGoogle Scholar
  19. Domingos-Lopes M, Stanton C, Ross P, Dapkevicius M, Silva C (2017) Genetic diversity, safety and technological characterization of lactic acid bacteria isolated from artisanal Pico cheese. Food Microbiol 63:178–190PubMedGoogle Scholar
  20. FAO/WHO (2001) Report of a joint FAO/WHO expert consultation on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. CórdobaGoogle Scholar
  21. Faye T, Tamburello A, Vegarud GE, Skeie S (2012) Survival of lactic acid bacteria from fermented milks in an in vitro digestion model exploiting sequential incubation in human gastric and duodenum juice. J Dairy Sci 95:558–566PubMedGoogle Scholar
  22. Flórez AB, Danielsen M, Korhonen J, Zycka J, von Wright A, Bardowski J, Mayo B (2007) Antibiotic survey of Lactococcus lactis strains to six antibiotics by Etest, and establishment of new susceptibility-resistance cut-off values. J Dairy Res 74(3):262–268PubMedGoogle Scholar
  23. Franciosi E, Settanni L, Cavazza A, Poznanski E (2009) Biodiversity and technological potential of wild lactic acid bacteria from raw cows' milk. Int Dairy J 19:3–11Google Scholar
  24. G-Alegría E et al (2004) High tolerance of wild Lactobacillus plantarum and Oenococcus oeni strains to lyophilisation and stress environmental conditions of acid pH and ethanol. FEMS Microbiol Lett 230:53–61PubMedGoogle Scholar
  25. Gurses M, Erdogan A (2006) Identification of lactic acid bacteria isolated from Tulum cheese during ripening period. Int J Food Prop 9:551–557Google Scholar
  26. Hoque M, Akter F, Hossain K, Rahman M, Billah M, Islam K (2010) Isolation, identification and analysis of probiotic properties of Lactobacillus spp. from selective regional yoghurts. World J Dairy Food Sci 5:39–46Google Scholar
  27. Hviid A-MM, Ruhdal-Jensen P, Kilstrup M (2017) Butanol is cytotoxic to Lactococcus lactis while ethanol and hexanol are cytostatic. Microbiol 163:453–461Google Scholar
  28. Karakas-Sen A, Karakas E (2018) Isolation, identification and technological properties of lactic acid bacteria from raw cow milk. Biosci J 34:385–399Google Scholar
  29. Khemariya P, Singh S, Nath G, Gulati AK (2013) Isolation, identification and antibiotic susceptibility of nis+ Lactococcus lactis from dairy and non-dairy sources. Czech J Food Sci 31(4):323–331Google Scholar
  30. Kimoto H, Kurisaki J, Tsuji N, Ohmomo S, Okamoto T (1999) Lactococci as probiotic strains: adhesion to human enterocyte-like Caco-2 cells and tolerance to low pH and bile. Lett Appl Microbiol 29:313–316PubMedGoogle Scholar
  31. Kondrotiene K et al (2018) Characterization and application of newly isolated nisin producing Lactococcus lactis strains for control of Listeria monocytogenes growth in fresh cheese. LWT-Food Sci Technol 87:507–514Google Scholar
  32. 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 94:981–987PubMedGoogle Scholar
  33. Kumar BV, Vijayendra SVN, Reddy OVS (2015) Trends in dairy and non-dairy probiotic products-a review. J Food Sci Technol 52:6112–6124Google Scholar
  34. Li S et al (2012) Antioxidant activity of Lactobacillus plantarum strains isolated from traditional Chinese fermented foods. Food Chem 135:1914–1919PubMedGoogle Scholar
  35. Lopez-Dıaz T, Alonso C, Roman C, Garcıa-Lopez M, Moreno B (2000) Lactic acid bacteria isolated from a hand-made blue cheese. Food Microbiol 17:23–32Google Scholar
  36. Meira SMM, Helfer VE, Velho RV, Lopes FC, Brandelli A (2012) Probiotic potential of Lactobacillus spp. isolated from Brazilian regional ovine cheese. J Dairy Res 79:119–127PubMedGoogle Scholar
  37. Mishra V, Prasad D (2005) Application of in vitro methods for selection of Lactobacillus casei strains as potential probiotics. Int J Food Microbiol 103:109–115PubMedGoogle Scholar
  38. Obis D, Guillot A, Mistou MY (2001) Tolerance to high osmolality of Lactococcus lactis subsp. lactis and cremoris is related to the activity of a betaine transport system. FEMS Microbiol Lett 202:39–44PubMedGoogle Scholar
  39. Ooi L-G, Liong M-T (2010) Cholesterol-lowering effects of probiotics and prebiotics: a review of in vivo and in vitro findings. Int J Mol Sci 11:2499–2522PubMedPubMedCentralGoogle Scholar
  40. Patel A, Lindström C, Patel A, Prajapati J, Holst O (2012) Probiotic properties of exopolysaccharide producing lactic acid bacteria isolated from vegetables and traditional Indian fermented foods. Int J Fermented Foods 1:87–101Google Scholar
  41. Patrick WA, Wagner HB (1949) Determination of hydrogen peroxide in small concentrations. Anal Chem 21:1279–1280Google Scholar
  42. Piard J, Desmazeaud M (1992) Inhibiting factors produced by lactic acid bacteria. 2. Bacteriocins and other antibacterial substances. Lait 72:113–142Google Scholar
  43. Picon A, Garde S, Ávila M, Nuñez M (2016) Microbiota dynamics and lactic acid bacteria biodiversity in raw goat milk cheeses. Int Dairy J 58:14–22Google Scholar
  44. Prasad J, Gill H, Smart J, Gopal PK (1998) Selection and characterisation of Lactobacillus and Bifidobacterium strains for use as probiotics. Int Dairy J 8:993–1002Google Scholar
  45. Randazzo CL, Vaughan EE, Caggia C (2006) Artisanal and experimental pecorino Siciliano cheese: microbial dynamics during manufacture assessed by culturing and PCR–DGGE analyses. Int J Food Microbiol 109:1–8PubMedGoogle Scholar
  46. Ribeiro S, Coelho M, Todorov S, Franco B, Dapkevicius M, Silva C (2014) Technological properties of bacteriocin-producing lactic acid bacteria isolated from Pico cheese an artisanal cow's milk cheese. J Appl Microbiol 116:573–585PubMedGoogle Scholar
  47. Rochat T, Miyoshi A, Gratadoux J, Duwat P, Sourice S, Azevedo V, Langella P (2005) High-level resistance to oxidative stress in Lactococcus lactis conferred by Bacillus subtilis catalase KatE. Microbiol 151:3011–3018Google Scholar
  48. Rodgers S (2008) Novel applications of live bacteria in food services: probiotics and protective cultures. Trends Food Sci Technol 19:188–197Google Scholar
  49. Sabir F, Beyatli Y, Cokmus C, Onal-Darilmaz D (2010) Assessment of potential probiotic properties of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains isolated from kefir. J Food Sci 75:M568–M573PubMedGoogle Scholar
  50. Shehata M, El Sohaimy S, El-Sahn MA, Youssef M (2016) Screening of isolated potential probiotic lactic acid bacteria for cholesterol lowering property and bile salt hydrolase activity. Ann Clin Lab Sci 61:65–75Google Scholar
  51. Solem C, Dehli T, Jensen PR (2013) Rewiring Lactococcus lactis for ethanol production. J Appl Environ Microbiol 79:2512–2518Google Scholar
  52. Takeda S et al (2011) The investigation of probiotic potential of lactic acid bacteria isolated from traditional Mongolian dairy products. Animal Sci J 82:571–579Google Scholar
  53. Tambekar D, Bhutada S (2010) Studies on antimicrobial activity and characteristics of bacteriocins produced by Lactobacillus strains isolated from milk of domestic animals. Internet J Microbiol 8:1–6Google Scholar
  54. Tulumoğlu Ş, Kaya Hİ, Şimşek Ö (2014) Probiotic characteristics of Lactobacillus fermentum strains isolated from tulum cheese. Anaerobe 30:120–125PubMedGoogle Scholar
  55. Umer Khan S (2014) Probiotics in dairy foods: a review Nutrition & Food Science 44:71-88Google Scholar
  56. Van Geel-Schutten G, Flesch F, Ten Brink B, Smith M, Dijkhuizen L (1998) Screening and characterization of Lactobacillus strains producing large amounts of exopolysaccharides. Appl Microbiol Biotechnol 50:697–703Google Scholar
  57. Vidhyasagar V, Saraniya A, Jeevaratnam K (2013) Identification of pectin degrading lactic acid bacteria from fermented food sources. Int J Adv Lif Sci 6:8–12Google Scholar
  58. Vinderola C, Reinheimer J (2003) Lactic acid starter and probiotic bacteria: a comparative “in vitro” study of probiotic characteristics and biological barrier resistance. Food Res Int 36:895–904Google Scholar
  59. Weichselbaum E (2009) Probiotics and health: a review of the evidence. Nutr Bull 34:340–373Google Scholar
  60. Yüksekdağ Z, Beyatli Y, Aslim B (2004) Determination of some characteristics coccoid forms of lactic acid bacteria isolated from Turkish kefirs with natural probiotic. LWT-Food Sci Technol 37:663–667Google Scholar

Copyright information

© Università degli studi di Milano 2019

Authors and Affiliations

  1. 1.Department of Food EngineeringUniversity of SelcukKonyaTurkey
  2. 2.Department of Food EngineeringKonya Food and Agriculture UniversityKonyaTurkey

Personalised recommendations