Identification and Characterization of <i>Lactococcus lactis ssp.lactis</i> Isolated from Raw Milk and Milk Products of Turkey and Iran


  • Shila Vahabzadeh Istanbul Aydin Univeristy (Istanbul,Turkey)
  • Haydar Özpınar Istanbul Aydin Univeristy (Istanbul,Turkey)
  • Indrani Kalkan Istanbul Aydin Univeristy (Istanbul,Turkey)
  • Serap Andaç Öztürk Istanbul Aydin Univeristy (Istanbul,Turkey)


Lactic acid bacteria, L.lactis ssp.lactis, starter culture, dairy products, antimicrobial, antibiotic


Lactococcus lactis ssp.lactis strains are the best known and characterized species of lactic acid bacteria which plays an important role as starter cultures in industrial dairy fermentation, owing to its pH lowering effect, production of antimicrobial metabolites and enhancement of the organoleptic attributes. The objective of this study was to identify and evaluate the varied properties of the natural L.lactis ssp.lactis strains from raw milk and dairy products obtained from different provinces of Turkey and Iran during the period of April to June 2016. Out of 262 dairy samples, L.lactis ssp.lactis were isolated from 98 samples. In order to evaluate the obtained strain as a good quality starter culture, litmus milk reaction, total titratable acidity, skim milk agar, antimicrobial activity and antibiotic resistance tests were conducted. The genetic basis of the observed phenotypic resistance obtained by disc diffusion test was further confirmed by PCR. Lactic acid production by Iranian cheese samples were significantly higher than Turkish samples (p<0.05) although difference between their proteolytic activities were statistically insignificant (p>0.05). Iranian cheese isolates showed higher antimicrobial activity againts E. faecalis and E. coli as compared to Turkish isolates. (p<0.05).  Difference in antibiotic resistance between Turkish and Iranian cheese samples were not statistically significant (p>0.05). However, tetracycline and streptomycin resistance gene was only confirmed by PCR in Turkish cheese samples. Emphasis must be given to selection of the milk source for obtaining good quality starter cultures.  Further studies may be conducted to learn the effect of ecological parameters on bacterial strains.


Author Biographies

Shila Vahabzadeh, Istanbul Aydin Univeristy (Istanbul,Turkey)

Department of Food Engineering

Haydar Özpınar, Istanbul Aydin Univeristy (Istanbul,Turkey)

Department of Food Engineering

Indrani Kalkan, Istanbul Aydin Univeristy (Istanbul,Turkey)

Department of Nutrition and Dietetics

Serap Andaç Öztürk, Istanbul Aydin Univeristy (Istanbul,Turkey)

Department of Nutrition and Dietetics


Atanasova J, Moncheva P, Ivanova I, “Proteolytic and antimicrobial activity of lactic acid bacteria grown in goat milkâ€, Biotechnology & Biotechnological Equipment,vol.28, no.6, pp. 1073-1078, 2014.

Aziz T, Khan H, Bakhtair SM, Naurin M, “Incidence and relative abundance of lactic acid bacteria in raw milk of buffalo,cow and sheepâ€,The Journal of Animal & Plant Sciences, vol.19, no.4, pp. 168-173, 2009.

Aziz KhE, Abdulraman ZFA, Essa RH, “Antibiotic Resistance pattern and Effect of Some Growth Condition on Lactococcus lactis subsp.lactis Isolated from Cow Milkâ€, Int.J.Curr.Microbiol, vol.4, no.6, pp. 388-405, 2015.

Bernardeau M, Vernoux JP, Henri-Dubernet S, Gueguen M, “Safety assessment of dairy microorganisms:The Lactobacillus genusâ€, Journal of Food Microbiology, vol.126, pp.278-285, 2008.

Bluma A, Ciprovica I, “Diversity of lactic acid bacteria in raw milkâ€, Research for rural development,vol.1, pp.157-161, 2015.

Boziaris IS, Humpheson L, Adams MR, “Effect of nisin on heat injury and inactivation of Salmonella enteritidis PT4â€, International Journal of Food Microbiology, vol.43, pp.7-13, 1998.

Boziaris IS, Adams MR,“Transient sensitivity to nisin in cold shocked Gram negativesâ€, Letters in Applied Microbiology, vol.31, pp.233-237, 2000.

Buyukyoruk S, Cibik R, Cetinkaya F, Soyutemis GE, Goksoy EO, Kirkan S, “Isolation,phenotypic and molecular identification of Lactococcus lactis isolation from traditionally produced village cheesesâ€, Journal of Animal and veterinary advanced, vol.9, no.16, pp.2154-2158, 2010.

Compaoré CS, Jensen LB, Diawara B, Ouédraogo GA, Jakobsen M, Ouoba Lll, “Resistance to antimicrobials and acid and bile tolerance of Bacillus spp isolated from Bikalga,fermented seeds of Hibiscus sabdariffaâ€, African J Food Sci, vol.7, no.11, pp.408-414, 2013.

Dal Bello D, Cocolin L, Zepa G, Field D, Cotter PD, Hill C, “Technological characterization of bacteriocin producing Lactococcus lactis strains employed to control Listeria monocytogenes in cottage cheeseâ€, Int.J.Food Microbiol, vol.153, pp.58-65, 2012.

Devirgilis C, Barile S, Caravelli A, Coppola D, Perozzi G, “Identification of Tetracycline and erythromycin-resistant Gram-positive cocci within the fermenting microflora of an Italian dairy food productâ€, Journal of Applied Microbiology, vol.109, pp.313-323, 2010.

Edalatian MR, Habibi MB, Mortazavi SA, Nasiri MR, Basami MR, Hashemi M, “Isolation and Identification the Indigenous Lactic Flora From Lighvan, as an Iranian Raw Milk Cheese From Milk to Ripened cheeseâ€, World Academy of Science, Engineering and Technology, vol.68, pp.1346-1351, 2010

El-Hassayeb HEA, AbdelAziz SMZ, “Screening, Production and Industrial Application of Protease Enzyme from Marine Bacteriaâ€, Int.J.Curr.Microbiol.App. Sci, vol.5, no.7, pp.863-874, 2016.

Enan G, Abdel-Shafi S, Ouda S, Negm S, “Novel Antibacterial Activity of Lactococcus Lactis Subspecies Lactis Z11 Isolated from Zabadyâ€, International journal of Biomedical science, vol.9, no.3, pp.174-180, 2013.

Facklam R, Elliott JA, “Identification,classification, and clinical relevance of catalase-negative,gram-positive cocci, excluding the streptococci and enterococciâ€, American Society for Microbiology ,Clinical Microbiology Reviews, vol.8, no.4, pp.479-495, 1995.

Feutry F, Torre P, Arana I, Garcia S, Desmasures N, Casalta E, “Lactococcus lactis strains from raw ewe's milk samples from the PDO Ossau-Iraty cheese area: levels, genotypic and technological diversityâ€, Dairy Science & Technology, vol.92, no.6, pp.655-670, 2012.

Florez AB, Danielsen M, Korhonen J, Zycka J, “Antibiotic survey of lactococcus lactis strains to six antibiotics by E-test and establishment of new susceptibility-resisrance cut-off valuesâ€, Journal of Dairy reaearch, vol.74, pp.262-268, 2007.

Gad GFM, Abdel-Hamid AM, Farag ZSH, “Antibiotic resistance in lactic acid bacteria isolated from some pharmaceutical and dairy productsâ€, Brazilian journal of Microbiology,vol.45, no.1, pp.25-33, 2014.

Gemechu T, “Review on lactic acid bacteria function in milk fermentation and preservationâ€, African Journal of Food Science, vol.9, no.4, pp.170-1749 ,2015.

Gevers D, Danielsen M, Huys G, Swings J, “Molecular characterization of tet(M)genes in Lactobacillus isolates from different types of fermented dry sausageâ€, Applied and Environmental Microbiology, vol.69, pp.1270-1275, 2003.

Gomez NC, Ramiro JMP, Quecan BXV, De melo Franco BDG, “use of potential probiotic lactic acid bacteria(LAB) biofilms for the control of listeria monocytogenes,salmonella typhimurium,and escherchia coli O157:H7 biofilms formationâ€, Frontiers in microbiology, vol.7, Article.863, 2016.

Guessas B, Kihal M, “Characterization of lactic acid bacteria isolated from Algerian arid zone raw goats' milkâ€, African Journal of Biotechnology, vol.3, no.6, pp.339-342, 2004.

Gutiérrez-Méndez N, Rodríguez-Figueroa JC, González-Córdova AF, Nevárez-Moorillón, GV, Rivera-Chavira B, Vallejo-Cordoba B, “Phenotypic and Genotypic Characteristics of Lactococcus lactis Strains Isolated from Different Ecosystemsâ€, Canadian Journal of Microbiology,vol.56, no.5, pp.432-439, 2010.

Hladíková Z, Smetanková J, Greif G, Greifová M, “Antimicrobial activity of selected lactic acid cocci and production of organic acidsâ€, Acta Chimica Slovaca, vol.5, no.1, pp.80-85, 2012.

Kahala M, Maki M, Lehtovaara A, Tapanainen JM, Katiska R, Juuruskorpi M, Juhola J, Joutsjoki V, “characterization of starter lactic acid bacteria from the finnish fermented milk product viiliâ€, Journal of Appled Microbiology, vol.105, pp.1929-1938, 2008.

Kalchayanand N, Hanlin MB, Ray B, “Sublethal injury makes Gram negative and resistant Gram positive bacteria sensitive to the bacteriocins,pediocin AcH and nisinâ€, Letters in Applied Microbiology,vol.15, pp.239-243, 1992.

Khemariya P, Singh S, Nath G, Gulati Ak, “Isolation,identification,and antibiotic suscep¬tibility of nis+ Lactococcus lactis from dairy and non-dairy sourcesâ€, Czech J.Food Sci, vol.31, pp.323-331, 2013.

Kim SY, Kim JE, Lee KW, Lee HJ, “L.lactis ssp.lactis inhibits the proliferation of SNU-1 human stomach cancer cells through induction of G0/G1 cell cycle arrest and apoptosis via p53 and p21 expressionâ€, Annals of the New York Academy of Sciences, vol.1171, pp.270-275, 2009.

Klare I, Konstabel C, Werner G, Huys G, Vankerckhoven V, Kahlmeter G, Hildebra B, Muller-Bertling S, Witte W, Goossens H, “Antimi¬crobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for pro¬biotic or nutritional useâ€, Journal of Antimicrobial Chemo¬therapy, vol.59, pp.900-912, 2007.

Ma C, Zhang L, Ma D, Du M, Han X, Yi H, Zhang L, Feng Z, Zhang Y, Zhang Y, Song W, “Technological characterization of Lactobacilli isolated from Chinese artisanal fermented milksâ€, International Journal of Dairy Technology. Vol.65, pp.132-139, 2012.

Ortigosa M, Arizcun C, Torre P, Marfa Izco J, “Use of wild Lactobacillus strainsin an adjunct culture for a Roncal-type cheeseâ€, Journal of Dairy Research, vol.72, pp.168-178, 2005.

Özkalp B, Özdena B, Tuncer Y¸ Åžanlibaba P, Akçelik M, “Technological characterization of wild-type Lactococcus lactis strains isolated from raw milk and traditional fermented milk products in Turkeyâ€, INRA EDP Sciences.Lait, vol.87, pp.521-534, 2007.

Pu ZY, Dobos M, Limsowtin GKY, Powell IB, “Integrated polymerase chain reaction-based procedures for the detection and identification of species and subspecies of the Gram-positive bacterial genus Lactococcusâ€, Journal of Applied Microbiology, vol.93, pp.353-361, 2002.

Rodríguez-Figueroa JC, González-Córdova AF, Astiazaran-García H, Vallejo-Cordoba B, “Hypotensive and heart rate-lowering effects in rats receiving milk fermented by specific Lactococcus lactis strainsâ€, British Journal of Nutrition, vo.109, pp.827-833, 2013.

Sharma P, Tomar SK, Goswami P, Sangwan V, Singh R, “Antibiotic resistance among commercially available probioticsâ€, Food Research International, vol.57, pp.176-195, 2014.

Suzuki S, Ogo M, Koike T, Takada H, Newman B, “Sulfonamide and tetracycline resistance genes intotal and culturable bacterial assemblages in South African aquatic environmentsâ€, Frontiers in Microbiology, vol.6, Article 796, 2015.

Titouche Y, Hakem A, Houali K, Yabrir B, Malki O, Chergui A, Chenouf N, Yahiaoui S, Labiad M, Ghenim H, Kechih-bounar S, Chirila F, NadaÅŸ G, Fit NI, “Detection of Antibiotics Residues in Raw milk Produced in Freha Area(Tizi-Ouzou),Algeriaâ€, Veterinary Medicine,vol.70, no.1, pp.83-87, 2013.

Tsakalidou E, Papadimitriou K, “Non-Bovine milk and milk productsâ€, Academic Press, London, UK, 2016.

Tükel Ç, Akçelik M, “Identification of Lactoce Plasmids in Lactococcus lastic subsp. lactis Stainsâ€, Turk Journal of Biology, vol.24, pp.405-424, 2000.

Vernile A, Giammanco GM, Spano G, Beresford T, Fox PF, Massa S, “Genotipic characterization of lactic acid bacteria isolated from traditional Pecorino Siciliano Cheeseâ€, Diary Science and Technology, vol.88, pp.619-629, 2008.

Walstra P, Walstra P, Wouters JTM, Geurts TJ, “Dairy science and technologyâ€, CRC Press, New York, US, 2005.

Wegener HC, “Antibiotics in animal feed and their role in resistance developmentâ€, Current Opinion in Microbiology, vol.6, pp.439-445, 2003.

Zycka-krzesinska J, Boguslawska J, Aleksandrzak-Piekarczyk T, Jopek J, Bardowski JK, “Identification and characterization of tetracycline resistance in Lactococcus lactis isolated from Polish raw milk and fermented artisanal productsâ€, International Journal of Food Microbiology, vol.211, pp.134-141, 2015.




How to Cite

Vahabzadeh, S., Özpınar, H., Kalkan, I., & Andaç Öztürk, S. (2017). Identification and Characterization of <i>Lactococcus lactis ssp.lactis</i> Isolated from Raw Milk and Milk Products of Turkey and Iran. Asian Journal of Agriculture and Food Sciences, 5(4). Retrieved from