Sakarya’da geleneksel olarak üretilen bazı yöresel peynirlerinden izole edilen laktik asit bakterilerinin MALDI-TOF MS Yöntemi ile tanımlanması ve antibiyotik dirençlerinin belirlenmesi

Year 2018, Volume: 22 Issue: 3, 1055 - 1062, 01.06.2018

Eda Kılıç Kanak , Suzan Öztürk Yılmaz

https://doi.org/10.16984/saufenbilder.346212

Abstract

Son yıllarda yöresel peynirler
tüketici tarafından daha fazla talep edilmektedir. Ülkemizde lezzetli birçok
yöresel peynir vardır. Fermantasyon süreçlerinde dikkatle seçilmiş suşların
starter kültür olarak uygulanması, doğal ve sağlıklı bir ürün elde edilmesini
sağlar. Bu çalışmada amaç çeşitli yöresel peynirlerin üretiminde ürüne tat,
koku, yapı ve görünüm bakımından istenilen nitelikleri kazandıran farklı laktik
asit bakterilerini (LAB) tespit etmektir. Bu amaçla Sakarya’da geleneksel
peynirlerden LAB’leri izole edilip MALDI-TOF MS yöntemi ile tanımlanmıştır. Bu
çalışmada geleneksel yöntemle üretilmiş 3 adet peynir örneği ile çalışılmıştır.
MALDI-TOF MS analizi ile de 16 suş tür düzeyinde tanımlanarak; izolatlardan 5
tanesi Enterococcus faecalis, 4
tanesi Enterococcus faecium, 1 tanesi
Lactobacillus paracasei ve 6 tanesi Leuconostoc mesenteroides olarak bulunmuştur.

Keywords

Laktik asit bakterileri, MALDI-TOF MS, Yöresel peynirler

Identification of lactic acid bacteria isolated from traditionally produced local cheese by MALDI-TOF MS anddetermination of antibiotic resistants

Abstract

In recent years, local cheeses have been demanded more
by consumers. There is a delicious variety of regional cheeses in Turkey. The
compounds that produce lactic acid bacteria improve the texture in cheese and
contribute to the pleasant sensory profile of the final product. The purpose of
this study is to determine the different lactic acid bacteria and their
antibiotic resistance  that give the
desired qualities in terms of taste, odor, structure and appearance of the
product in the production of various regional cheeses. For this purpose lactic
acid bacteria were isolated from traditional cheeses in Sakarya and identified
by MALDI-TOF MS method. In this study, 3 cheese samples produced by traditional
methods were used. The 15 strains were identified atspecies levels as E.
faecalis (5), E. faecium (4), Lb. paracasei (1)
and Leu. mesenteroides (6) by MALDI-TOF MS analysis. Antibiotic
susceptibility of these 15 strains was investigated. Strains which related to
23 lactic acid bacteria have been found resistant despite of 40% vancomycin and
nitrofurantoine, 33% gentamycin, 20% erythromycin and tetracycline, 13%
rifampicin and chloramphenicol.

Keywords

Lactic acid bacteria 1, MALDI-TOF MS 2, local cheeses 3, antibiotic resistance 4

References

• [1] M. Eralp, ‘‘Peynir Teknolojisi,’’ Ankara Üniversitesi Basımevi, Ankara, 331, 1973.
• [2] A. K. Halkman and Y. Taşkın, ‘‘Ulusal düzeyde laktik starter kültür üretimi,’’ 1. Uluslararası Adriyatik’ten Kafkaslara Geleneksel Gıdalar Sempozyumu, 15-17 Nisan 2010.
• [3] F. Leroy and L. De Vuyst, ‘‘Lactic acid bacteria as functional starter cultures for the food fermentation industry,’’ Trends in Food Science and Technology, 15(2), 67-78, 2004.
• [4] F. Hillenkamp and J. Peter-Katalinic, ‘‘MALDI MS: A practical guide to ınstrumentation. ın: methods and applications,’’ John Wiley and Sons, 345, 2007.
• [5] A. Wieser, L. Schneider, J. Jung, S. Schubert, ‘‘MALDI-TOF MS in microbiological diagnosticsidentification of microorganisms and beyond,’’ Applied microbiology and biotechnology, 93(3), 965-974, 2012.
• [6] S. B. Barbuddhe, T. Maier, G. Schwarz, M. Kostrzewa, H. Hof, E. Domann, T. Chakraborty, T. Hain, ‘’Rapid identification and typing of listeria species by matrixassisted laser desorption ionization-time of flight mass spectrometry,’’ Applied and Environmental Microbiology, 74: 5402– 07, 2008.
• [7] A. Mellmann, J. Cloud, T. Maier U. Keckevoet, I. Ramminger, P. Iwen, J. Dunn, G. Hall, D. Wilson, P. LaSala, M. Kostrzewa, D. Harmsen, ‘’Evaluation of matrix-assisted laser desorption ionization-time-off light mass spectrometry in comparison to 16S rRNA gene sequencing for species identification of non fermenting bacteria,’’ Journal of clinical microbiology, 46(6), 1946-1954, 2008.
• [8] P. Seng, M. Drancourt, F. Gouriet, B. La Scola, P. E. Fournier, J. M. Rolain, D. Raoult, ‘’Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry,’’ Clinical Infectious Diseases, 49(4), 543-551, 2009.
• [9] G. Prod’hom, A. Bizzini, C. Durussel, J. Bille, G. Greub, ‘’Matrix assisted laser desorption ionization-time of flight mass spectrometry for direct bacterial identification from positive blood culture pellets,’’ Journal of clinical microbiology, 48(4), 1481-1483, 2010.
• [10] L. G. Stevenson S. K. Drake P. R. Murray, ‘’Rapid identification of bacteria in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry,’’ Journal of clinical microbiology, 48(2), 444-447, 2010.
• [11] A. Cherkaoui, J. Hibbs, S. Emonet, M. Tangomo, M. Girard, P. Francois, J. Schrenzel, ‘’Comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry methods with conventional phenotypic identification for routine identification of bacteria to the species level,’’ Journal of clinical microbiology, 48: 1169–75, 2010.
• [12] J. P. Anhalt, C. Fenselau, ‘’Identification of bacteria using mass spectrometry,’’ Analytical Chemistry. 47: 219-25, 1975.
• [13] M. J. Suh, P. A. Limbach, ‘’Investigation of methods suitable for the matrix-assisted laser desorption/ ionization mass spectrometric analysis of proteins from ribonucleoprotein complexes,’’ European Journal of Mass Spectrometry, 10(1), 89-100, 2004.
• [14] A. Temiz, Genel Mikrobiyoloji Uygulama Teknikleri. 3. Baskı. Hatipoğlu Yayınları, Ankara. 291, 2000.
• [15] S. W. Kim, L. Perl, H. J. Park, E. J. Tandianus, W. N. Dunn, W. N. ‘’Assesment of stress responce of the probiotic,’’ Lactobacillus acidophilus. Current Microbiology, 43: 346-350, 2001.
• [16] F. J. Carr, D. Chill, N. Miada,‘’The lactic acid bacteria: a literature survey,’’ Critical reviews in microbiology, 28(4), 281-370, 2002.
• [17] N. Özcan, Ö. Ezin, N. Akpolat, H. Bozdağ, M. Mete, K. Gül, ‘’Klinik örneklerde saptanan Candida türlerinin MALDI-TOF MS ile tiplendirilmesi,’’ Dicle Medical Journal 43 (3): 390-394, 2016.
• [18] D. Korucu, ‘’Tomas peynirinden izole edilen laktik asit bakterilerinin tanımlanması,’’ Yüksek Lisans Tezi, Namık Kemal Üniversitesi, Fen Bilimleri Enstitüsü, Tekirdağ, 32, 2012.
• [19] Z. Öner, O. Sağdiç, B. Şimşek, ‘’Lactic acid bacteria profiles and tyramine and tryptamine contents of Turkish tulum cheeses,’’ European Food Research and Technology, 219, 455-459, 2004.
• [20] Y. Tuncer, ‘’Some technological properties of phenotypically identified enterococci strains isolated from Turkish tulum cheese,’’ African Journal of Biotechnology, 8 (24), 7008-7016, 2009.
• [21] N. Yoğurtçu, ‘’Tulum peynirinden enterokok suşlarının izolasyonu ve antibiyotik duyarlılıklarının belirlenmesi,‘’ Yüksek Lisans Tezi, Süleyman Demirel Üniversitesi, Fen Bilimleri Enstitüsü, Isparta, 30-41, 2011.
• [22] A. C. Freitas, C. Paris, F. X. Malcata, T. A. Hogg, ‘’Microbiological characterization of Picante da Beira Baixa cheese,’’ Journal of Food Protection, 59; 155-160, 1996.
• [23] G. Suzzi, M. Caruso, F. Gadrini, A. Lombardi, L. Vanni, M. E. Guerzoni, C. Andrighetto, M. T. Lanorte, ‘’A survey of the enterococci isolated from an artisanal Italian goat’s cheese (semicotto caprino),’’ Journal of Applied Microbiology, 89, 267-274, 2000.
• [24] D. Jurkovič, L. Križkova´, R. Dušinsky´, A. Belicova´, M. Sojka, J. Krajčovič, L. Ebringer, ‘’Identification and characterization of enterococci from bryndza cheese,’’ Letters in Applied Microbiology, 42, 553-559, 2006.
• [25] M. Nicolic, A. Terzic-Vidojevic, B. Jovcic, J. Begovic, N. Golic, L. Topisirovic, ‘’Characterization of lactic acid bacteria isolated from Bukuljac, a homemade goat's milk cheese,’’ International Journal of Food Microbiology, 122: 162–170, 2008.
• [26] A. G. Williams, J. M. Banks, ‘’Proteolytic and other hydrolytic enzyme activities in non-starter lactic acid bacteria (nslab) ısolated from cheddar cheese manufactured in the United Kingdom,’’ Dairy Journal, 7: 763-774, 1997.
• [27] K. V. Hoorde, T. Verstraete, P. Vandamme, G. Huys, ‘’Diversity of lactic acid bacteria in two Flemish artisan raw milk Gouda-type cheeses,’’ Food Microbiology, 25: 929–935, 2008.
• [28] M. Sengül, S. Çakmakcı, ‘’Characterization of natural isolates of lactic acid bacteria from Erzincan (Savak) Tulum cheese,’’ Milchwissenschaft, 58(9-10), 510-513, 2003.
• [29] M. Gurses, A. Erdoğan, ‘’Identification of lactic acid bacteria isolated from Tulum cheese during ripening period,’’ International Journal of Food Properties (9): 551-557, 2006.