Molecular characterization of yeasts isolated from traditional Turkish cheeses

Sine OZMEN TOGAY Angela CAPECE Gabriella SIESTO Harun AKSU Sema SANDIKCI ALTUNATMAZ Filiz YILMAZ AKSU Patrizia ROMANO Yonca KARAGUL YUCEER About the authors

Abstract

Thirty-two yeast strains were identified by means of molecular methods isolated from traditional Turkish cheeses (Tulum, Kashkaval, Mihalic, Orgu, White, Sepet, and Goat). Debaryomyces hansenii and Torulaspora delbrueckii were found as predominant species in cheese samples. Other species which were identified were Kluyveromyces lactis, Candida parapsilosis, Clavispora lusitaniae, Saccharomyces cerevisiae, K. marxianus, Rhodotorula mucilaginosa, Meyerozyma guilliermondii (formerly Pichia guilliermondii), C. zeylanoides and Candida albicans. Rhodotorula mucilaginosa and D. hansenii strains, from Kashkaval cheese, showed antilisterial activity, whereas only one K. lactis strain from Orgu cheese exhibited proteolytic activity.

Keywords:
Tulum, Kashkaval, Mihalic, Orgu, White, Sepet, and Goat cheeses; Debaryomyces hansenii; Torulaspora delbrueckii; Kluyveromyces lactis; Rhodotorula mucilaginosa; characterization; antimicrobial activity

1 Introduction

Yeasts are an essential part of cheese microflora because they can tolerate low water activity and pH, high salt concentrations and low storage temperatures and are resistant to some chemicals such as sanitizers and cleaning compounds (Ferreira & Viljoen, 2003Ferreira, A. D., & Viljoen, B. C. (2003). Yeasts as adjunct starters in matured Cheddar cheese. International Journal of Food Microbiology, 86(1-2), 131-140. http://dx.doi.org/10.1016/S0168-1605(03)00252-6. PMid:12892928.
http://dx.doi.org/10.1016/S0168-1605(03)...
; Banjara et al., 2015Banjara, N., Suhr, M. J., & Hallen-Adams, H. E. (2015). Diversity of yeast and mold species from a variety of cheese types. Current Microbiology, 70(6), 792-800. http://dx.doi.org/10.1007/s00284-015-0790-1. PMid:25694357.
http://dx.doi.org/10.1007/s00284-015-079...
). Certain yeasts may have the ability to use lactose, proteins, lipids, and some organic acids and may be applied for ripening by generating some flavor components and modifying the texture of cheeses and dairy products. Traditional fermented dairy products such as kefir and kumys are also produced by lactic acid bacteria and yeast fermentation (Jakobsen & Narvhus, 1996Jakobsen, M., & Narvhus, J. (1996). Yeasts and their possible beneficial and negative effects on the quality of dairy products. International Dairy Journal, 6(8-9), 755-768. http://dx.doi.org/10.1016/0958-6946(95)00071-2.
http://dx.doi.org/10.1016/0958-6946(95)0...
; Vasdinyei & Deak, 2003Vasdinyei, R., & Deak, T. (2003). Characterization of yeast isolates originating from Hungarian dairy products using traditional and molecular identification techniques. International Journal of Food Microbiology, 86(1-2), 123-130. http://dx.doi.org/10.1016/S0168-1605(03)00251-4. PMid:12892927.
http://dx.doi.org/10.1016/S0168-1605(03)...
; Ferreira & Viljoen, 2003Ferreira, A. D., & Viljoen, B. C. (2003). Yeasts as adjunct starters in matured Cheddar cheese. International Journal of Food Microbiology, 86(1-2), 131-140. http://dx.doi.org/10.1016/S0168-1605(03)00252-6. PMid:12892928.
http://dx.doi.org/10.1016/S0168-1605(03)...
; Büchl & Seiler, 2011Büchl, N. R., & Seiler, H. (2011). Yeast and molds: yeasts in milk and dairy products. In H. Roginski, J. W. Fuquay, P. F. Fox, eds. Encyclopedia of dairy sciences (2nd ed., pp. 744-753). New York: Elsevier. http://dx.doi.org/10.1016/B978-0-12-374407-4.00498-2.
http://dx.doi.org/10.1016/B978-0-12-3744...
; Banjara et al., 2015Banjara, N., Suhr, M. J., & Hallen-Adams, H. E. (2015). Diversity of yeast and mold species from a variety of cheese types. Current Microbiology, 70(6), 792-800. http://dx.doi.org/10.1007/s00284-015-0790-1. PMid:25694357.
http://dx.doi.org/10.1007/s00284-015-079...
).

At the same time, some yeast strains can cause spoilage by producing gas, off-flavors, softening of texture and discoloration changes in dairy products, especially with low pH such as yogurt, cream cheese, fermented milk, sour cream, etc. (Vasdinyei & Deak, 2003Vasdinyei, R., & Deak, T. (2003). Characterization of yeast isolates originating from Hungarian dairy products using traditional and molecular identification techniques. International Journal of Food Microbiology, 86(1-2), 123-130. http://dx.doi.org/10.1016/S0168-1605(03)00251-4. PMid:12892927.
http://dx.doi.org/10.1016/S0168-1605(03)...
; Kavas et al., 2006Kavas, G., Kinik, O., Uysal, S., Kilic, S., Celikel, N., & Akbulut, N. (2006). Characterisation of yeasts isolated from artisanal Turkish dairy products. International Journal of Dairy Science, 1, 44-50. http://dx.doi.org/10.3923/ijds.2006.44.50.
http://dx.doi.org/10.3923/ijds.2006.44.5...
; Büchl & Seiler, 2011Büchl, N. R., & Seiler, H. (2011). Yeast and molds: yeasts in milk and dairy products. In H. Roginski, J. W. Fuquay, P. F. Fox, eds. Encyclopedia of dairy sciences (2nd ed., pp. 744-753). New York: Elsevier. http://dx.doi.org/10.1016/B978-0-12-374407-4.00498-2.
http://dx.doi.org/10.1016/B978-0-12-3744...
; Corbaci et al., 2012Corbaci, C., Ucar, F. B., & Yalcin, H. T. (2012). Isolation and characterization of yeasts associated with Turkish-style homemade dairy products and their potential as starter cultures. African Journal of Microbiological Research, 6, 534-542.).

Yeasts may enter cheese production process from a variety of sources, such as the starter microorganisms, brine, environmental air, workers, and processing equipment (Banjara et al., 2015Banjara, N., Suhr, M. J., & Hallen-Adams, H. E. (2015). Diversity of yeast and mold species from a variety of cheese types. Current Microbiology, 70(6), 792-800. http://dx.doi.org/10.1007/s00284-015-0790-1. PMid:25694357.
http://dx.doi.org/10.1007/s00284-015-079...
). The most commonly isolated yeast genera from dairy products are Debaryomyces, Yarrowia, Candida, Zygosaccharomyces, Cryptococcus, Geotrichum, Kluyveromyces, Trichosporon, Rhodotorula, Torulaspora, and Saccharomyces. Kluyveromyces marxianus, Debaryomyces hansenii, and Saccharomyces cerevisiae species are mostly presented in dairy samples. Besides, Yarrowia lipolytica and D. hansenii may also be used as adjunct cultures for flavor enhancement during cheese ripening (Hansen & Jakobsen, 2001Hansen, T. K., & Jakobsen, M. (2001). Taxonomical and technological characteristics of Saccharomyces spp. associated with blue-veined cheese. International Journal of Food Microbiology, 69(1-2), 59-68. http://dx.doi.org/10.1016/S0168-1605(01)00573-6. PMid:11589561.
http://dx.doi.org/10.1016/S0168-1605(01)...
; Ferreira & Viljoen, 2003Ferreira, A. D., & Viljoen, B. C. (2003). Yeasts as adjunct starters in matured Cheddar cheese. International Journal of Food Microbiology, 86(1-2), 131-140. http://dx.doi.org/10.1016/S0168-1605(03)00252-6. PMid:12892928.
http://dx.doi.org/10.1016/S0168-1605(03)...
; Vasdinyei & Deak, 2003Vasdinyei, R., & Deak, T. (2003). Characterization of yeast isolates originating from Hungarian dairy products using traditional and molecular identification techniques. International Journal of Food Microbiology, 86(1-2), 123-130. http://dx.doi.org/10.1016/S0168-1605(03)00251-4. PMid:12892927.
http://dx.doi.org/10.1016/S0168-1605(03)...
; Capece & Romano, 2009Capece, A., & Romano, P. (2009). “Pecorino di Filiano” cheese as a selective habitat for the yeast species, Debaryomyces hansenii, Short communication. International Journal of Food Microbiology, 132(2-3), 180-184. http://dx.doi.org/10.1016/j.ijfoodmicro.2009.04.007. PMid:19411124.
http://dx.doi.org/10.1016/j.ijfoodmicro....
).

Turkish White cheese, Kashkaval, and Tulum cheeses are the best known and the most commonly consumed cheese varieties which have a national and economic value in Turkey. Mihalic, Orgu, Sepet, and Goat cheese are the other crucial Turkish cheese varieties (Hayaloglu et al., 2002Hayaloglu, A. A., Guven, M., & Fox, P. F. (2002). Microbiological, biochemical and technological properties of Turkish white cheese ‘Beyaz Peynir’ Review. International Dairy Journal, 12(8), 635-648. http://dx.doi.org/10.1016/S0958-6946(02)00055-9.
http://dx.doi.org/10.1016/S0958-6946(02)...
; Kamber, 2008Kamber, U. (2008). The traditional cheeses of Turkey: cheeses common to all regions. Food Reviews International, 24(1), 1-38. http://dx.doi.org/10.1080/87559120701761833.
http://dx.doi.org/10.1080/87559120701761...
; Aday & Karagul Yuceer, 2014Aday, S., & Karagul Yuceer, Y. (2014). Physicochemical and sensory properties of Mihalic cheese. International Journal of Food Properties, 17(10), 2207-2227. http://dx.doi.org/10.1080/10942912.2013.790904.
http://dx.doi.org/10.1080/10942912.2013....
).

In the researches about yeast microflora of Turkish cheeses, D. hansenii, Pichia amethionina var. amethionina, K. lactis and Candida spp., were identified in Turkish White cheeses (Hayaloglu et al., 2002Hayaloglu, A. A., Guven, M., & Fox, P. F. (2002). Microbiological, biochemical and technological properties of Turkish white cheese ‘Beyaz Peynir’ Review. International Dairy Journal, 12(8), 635-648. http://dx.doi.org/10.1016/S0958-6946(02)00055-9.
http://dx.doi.org/10.1016/S0958-6946(02)...
); Candida, Geotrichum, Kluyveromyces, Pichia, Saccharomyces and Zygosaccharomyces genera were found in Tulum cheeses (Karasu-Yalcin et al., 2012Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2012). Identification and enzymatic characterization of the yeasts isolated from Erzincan Tulum cheese. Mljekarstvo, 62, 53-61.); Debaryomyces spp., Candida spp, Kluyveromyces spp., Trichosporon spp., Saccharomyces spp. and Geotricum spp. were obtained from Tulum, white pickled and, Kasar (Kashkaval) cheeses (Kavas et al., 2006Kavas, G., Kinik, O., Uysal, S., Kilic, S., Celikel, N., & Akbulut, N. (2006). Characterisation of yeasts isolated from artisanal Turkish dairy products. International Journal of Dairy Science, 1, 44-50. http://dx.doi.org/10.3923/ijds.2006.44.50.
http://dx.doi.org/10.3923/ijds.2006.44.5...
); Candida (especially C. famata var. famata), Geotrichum and Trichosporon spp. were identified as dominant yeast flora in Mihalic cheeses (Karasu-Yalcin et al., 2017Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2017). Enzymatic characterization of yeast strains originated from traditional Mihalic cheese. Journal of Microbiology, Biotechnology and Food Sciences, 6(5), 1152-1156. http://dx.doi.org/10.15414/jmbfs.2017.6.5.1152-1156.
http://dx.doi.org/10.15414/jmbfs.2017.6....
); and D. hansenii was found as predominant yeast species in homemade cheeses (Corbaci et al., 2012Corbaci, C., Ucar, F. B., & Yalcin, H. T. (2012). Isolation and characterization of yeasts associated with Turkish-style homemade dairy products and their potential as starter cultures. African Journal of Microbiological Research, 6, 534-542.).

It has also been mentioned that certain species of yeasts may have inhibition characteristics against spoilage or pathogenic microorganisms in cheeses. D. hansenii may have antibacterial activity against Clostridium butyricum and C. tyrobutyricum in cheese brines (Ferreira & Viljoen, 2003Ferreira, A. D., & Viljoen, B. C. (2003). Yeasts as adjunct starters in matured Cheddar cheese. International Journal of Food Microbiology, 86(1-2), 131-140. http://dx.doi.org/10.1016/S0168-1605(03)00252-6. PMid:12892928.
http://dx.doi.org/10.1016/S0168-1605(03)...
). The antimicrobial activity of yeasts has been known for years. It is thought that antagonistic effects of yeasts are obtained through competition for nutrients, production of ethanol and organic acid, antimicrobial compounds such as extracellular proteins or glycoproteins called mycocins (Hatoum et al., 2013Hatoum, R., Labrie, S., & Fliss, I. (2013). Identification and partial characterization of antilisterial compounds produced by dairy yeasts. Probiotics and Antimicrobial Proteins, 5(1), 8-17. http://dx.doi.org/10.1007/s12602-012-9109-8. PMid:26782600.
http://dx.doi.org/10.1007/s12602-012-910...
; Buyuksirit & Kuleasan, 2014Buyuksirit, T., & Kuleasan, H. (2014). Antimicrobial agents produced by yeasts. World Academy of Science, Engineering and Technology International Journal of Biological, Veterinary, Agricultural, and Food Engineering, 8, 999-1002.). Certain yeast genera, such as Saccharomyces, Debaryomyces, Cryptococcus, Candida, Hanseniaspora, Kluyveromyces, Pichia, Torulopsis, Williopsis and Zygosaccharomyces, may produce mycocins (Hatoum et al., 2012Hatoum, R., Labrie, S., & Fliss, I. (2012). Antimicrobial and probiotic properties of yeasts: from fundamental to novel applications. Frontiers in Microbiology, 3 (421). http://dx.doi.org/10.3389/fmicb.2012.00421.
http://dx.doi.org/10.3389/fmicb.2012.004...
; Buyuksirit & Kuleasan, 2014Buyuksirit, T., & Kuleasan, H. (2014). Antimicrobial agents produced by yeasts. World Academy of Science, Engineering and Technology International Journal of Biological, Veterinary, Agricultural, and Food Engineering, 8, 999-1002.).

In this research, the wild yeast strains isolated from traditional Turkish cheeses were identified by means of molecular techniques. The antagonistic effect and the proteolytic and lipolytic activities of the isolates were also evaluated.

2 Materials and methods

2.1 Yeast isolation

The cheese samples belonging to Tulum, Kashkaval, Mihalic, Orgu, White, Sepet, and Goat cheese varieties were collected (n = 50). Samples (25g) were weighed in sterile bags and homogenized in 225 mL buffered peptone water using a peristaltic blender (Stomacher, ISOLAB, Turkey). The decimal dilutions of the samples were spread on YEPD (yeast extract–peptone–dextrose) agar (Merck, Darmstadt, Germany) medium with 0.1 g/L chloramphenicol (Merck, Darmstadt, Germany) added and incubated during 2-6 days at 28 °C. One to three colonies were selected randomly from the YEPD agar, subcultured to obtain pure cultures on YEPD medium (Capece & Romano, 2009Capece, A., & Romano, P. (2009). “Pecorino di Filiano” cheese as a selective habitat for the yeast species, Debaryomyces hansenii, Short communication. International Journal of Food Microbiology, 132(2-3), 180-184. http://dx.doi.org/10.1016/j.ijfoodmicro.2009.04.007. PMid:19411124.
http://dx.doi.org/10.1016/j.ijfoodmicro....
).

2.2 Yeast identification

Genomic DNA extraction was performed using a synthetic resin (Instagene Bio-Rad Matrix), following the protocol described by Capece et al. (2011)Capece, A., Pietrafesa, R., & Romano, P. (2011). Experimental approach for target selection of wild wine yeasts from spontaneous fermentation of “Inzolia” grapes. World Journal of Microbiology & Biotechnology, 27(12), 2775-2783. http://dx.doi.org/10.1007/s11274-011-0753-z.
http://dx.doi.org/10.1007/s11274-011-075...
. The presumptive identification of the yeasts obtained from traditional Turkish cheeses was performed by PCR amplification of the internal transcribed spacers between the 18S and 26S rDNA genes (ITS1-5.8S-ITS2) and subsequent restriction analysis, as reported by Esteve-Zarzoso et al. (1999)Esteve-Zarzoso, B., Belloch, C., Uruburu, F., & Querol, A. (1999). Identification of yeasts by RFLP analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers. International Journal of Systematic Bacteriology, 49(Pt 1), 329-337. PMid:10028278.. PCR products were digested without further purification with restriction enzymes HaeIII and HinfI (Promega, USA). Restricted fragments were analyzed by electrophoresis on 2% agarose gels, and 1.0X TBE buffer, stained with SyberSafe DNA gel stain (Invitrogen, USA) and the obtained profiles were visualized and photographed with Gel DocTM XR+ (Bio-Rad, USA) under UV light. A 100-bp DNA ladder marker (Promega, USA) served as the standard size. Isolates showing the same restriction pattern were grouped, and one or two representative yeasts for each group were delivered to Eurofins Genomics Srl (Vimodrone, Italy) for sequencing. The sequences received were compared with those deposited in the GenBank DNA database (National Center for Biotechnology Information, 2015National Center for Biotechnology Information – NCBI. (2015). Bethesda: NCBI. Retrieved from http://www.ncbi.nlm.nih.gov
http://www.ncbi.nlm.nih.gov...
) using the basic BLAST search tools (Altschul et al., 1997Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W., & Lipman, D. J. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research, 25(17), 3389-3402. http://dx.doi.org/10.1093/nar/25.17.3389. PMid:9254694.
http://dx.doi.org/10.1093/nar/25.17.3389...
).

2.3 Molecular characterization using MSP-PCR

The genetic variability among the analyzed strains was evaluated by means of microsatellite-primed PCR (MSP-PCR) experiments. The primers employed in MSP-PCR assays were the minisatellite M13 (GAGGGTGGCGGTTCT) and the microsatellite synthetic oligonucleotide (GTG)5. Amplification reactions were done in a total volume of 50 µL containing 10 µL of Taq Polymerase 5X Buffer (Promega, USA), 4.0 µL of 25 mM MgCl2 (Promega, USA), 1 µL of 10 mM dNTP (Promega, USA), 5 µL of 5 µM primer, 0.25 µL (5 U/µL) of Taq DNA polymerase (Promega, USA) and 5 µL of the extracted DNA, by adding sterile water until final volume. The thermal cycler was programmed as follows: initial denaturation at 95 °C for 5 min, followed by 35 cycles at 94 °C for 1 min, 1 min at 52 °C, 2 min at 72 °C for (GTG)5, while the initial denaturation was followed by 40 cycles at 95 °C for 40 sec, 1 min at 52 °C, 1 min at 72 °C; the final extension step for both primers was at 72 °C for 5 min for M13. The amplification was conducted in duplicate for each sample to assess the reproducibility of the obtained patterns. The PCR products were determined by agarose gel electrophoresis, as previously described. The conversion, normalization and further analysis of the patterns were carried out with FP QuestTM software ver. 4.5 (Bio-Rad, USA).

2.4 Antagonistic effect of the yeast strains

The antagonistic effect of the yeast strains against Listeria monocytogenes ATCC 7644, Staphylococcus aureus ATCC 6538 and Enterococcus faecalis ATCC 29212 were evaluated by using agar spotting method.

The 3 µL of 24 h cultures of the yeast isolates were spotted onto the surface of YEPD agar and incubated during 48-72 h at 30 °C. Then, the plates were covered by ten mililiter of BHI soft agar (0.7% agar) inoculated with 10 µL of one of the Listeria monocytogenes ATCC 7644, Staphylococcus aureus ATCC 6538, and Enterococcus faecalis ATCC 29212 test cultures. After incubation during 18-24 hours at 37 °C, the test plates were checked for clear inhibition zone diameters at around test cultures (Harris et al., 1989Harris, L. J., Daeschel, M. A., Stiles, M. E., & Klaenhammer, T. R. (1989). Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes. Journal of Food Protection, 52(6), 384-387. http://dx.doi.org/10.4315/0362-028X-52.6.384. PMid:31003307.
http://dx.doi.org/10.4315/0362-028X-52.6...
). The antagonistic effects of yeasts were also confirmed using neutralized supernatants (supernatant pH adjusted as 7.0) from the cultures.

2.5 Proteolytic and lipolytic activity of the yeast strains

Skim milk agar (Oxoid, UK) was used for determination of proteolytic activity of yeasts. Lipolytic activity was analyzed by spotting 24 h cultures of the yeasts on Tributyrin Agar (Oxoid, UK) and incubating at 30 °C (Lanciotti et al. 2004Lanciotti, R., Chaves-López, C., Patrignani, F., Paparella, A., Guerzoni, M. E., Serio, A., & Suzzi, G. (2004). Effects of milk treatment with dynamic high pressure on microbial populations, and lipolytic and proteolytic profiles of Crescenza cheese. International Journal of Dairy Technology, 57(1), 19-25. http://dx.doi.org/10.1111/j.1471-0307.2004.00121.x.
http://dx.doi.org/10.1111/j.1471-0307.20...
).

3 Results

3.1 Isolation and characterization of yeasts

The yeast loads were between 1.47 and 4.71 log cfu g-1 in 24 of 50 (48%) analyzed cheese samples in the study (Table 1). The yeast counts were of 2.87-4.71 log cfu g-1 for Tulum, 1.47-3.81 log cfu g-1 for Kashkaval, 3.61-4.36 log cfu g-1 for Mihalic, 2.68-4.71 log cfu g-1 for Kelle, 3.44-4.17 log cfu g-1 for Orgu, 3.39 log cfu g-1 for White, 2.91 log cfu g-1 for Sepet and 4.53 log cfu g-1 for Goat cheese samples. (Table 1). Thirty-two yeast strains isolated from Turkish cheeses in the present study were ascribed to 11 different species by RFLP analysis and sequencing of ITS region. Table 2 shows the size of digested PCR products with HaeIII and HinfI and the results of ITS sequencing, performed on representatives of different RFLP profiles found. All the sequenced yeast isolates showed high similarity (100% for all the isolates, except those of Rhodotorula mucilaginosa) to the sequences in Genbank (Table 2). The most frequent species was, as expected, Debaryomyces hansenii, with 16 isolates belonging to this species, followed by Torulaspora delbrueckii (3 isolates), whereas the species K. lactis, C. parapsilosis and Clavispora lusitaniae were present with two isolates for each one. Other species, such as S. cerevisiae, K. marxianus, Rhodotorula mucilaginosa, Meyerozyma guilliermondii (formerly Pichia guilliermondii), C. zeylanoides and C. albicans, were represented by only one isolate (Table 2).

Table 1
The yeast counts and number of positive cheese samples.
Table 2
Molecular identification results of yeasts isolated from Turkish cheeses.

3.2 Molecular characterization using MSP-PCR

Based on the identification results, yeast species including at least two isolates were submitted to molecular characterization at the strain level. The genetic polymorphism was analyzed using MSP-PCR with M13 and (GTG)5 primers.

The numerical analysis (UPGMA clustering) of the combined MSP-PCR patterns with the two oligonucleotides resulted in the dendrogram reported in Figure 1. The reproducibility of the MSP-PCR was higher than 90% (data not shown). Clusters were arbitrarily identified at a similarity level of 60%. The isolates were grouped in 6 main clusters and one single strain (56-2). The analyzed isolates were grouped according to the species, except D. hansenii isolates, which were separated into two clusters, 1 and 4. Cluster 1 was composed of one biotype, whereas cluster 4 could be further subdivided (at a similarity level of about 85%) in three different subclusters, 4a, 4b, and 4c, plus two single strains (49-1 and 19-1), indicating a significant polymorphism among D. hansenii strains. Certain biodiversity was also found in T. delbrueckii; in fact, one of the four strains grouped separately (56-2). The 56-1 and 56-2 strains were not cheese-derived. The others are included in cluster 5, in which two biotypes can be individuated, the first one, shared by the isolates 34-1 and 39-1, and the other one, exhibited by 47-1. The two Cl. lusitaniae strains (cluster 3) showed different biotypes, as did the two K. lactis strains, composing cluster 6. Otherwise, the two C. parapsilosis isolates, belonging to cluster 2, showed the same biotype. MSP-PCR revealed a remarkably high genetic diversity within our set of strains; however, the low correlation with the source of isolation was found.

Figure 1
Cluster analysis of the profiles obtained by means of MSP-PCR with M13 and (GTG)5 oligonucleotides (PD 30-1; D. hansenii, PD 40-2; D. hansenii, PD 45-1; D. hansenii, PD 21-1; C. parapsilosis, PD 28-1; C. parapsilosis, PD 25-1; Clavispora lusitaniae, PD 56-1; Clavispora lusitaniae, PD 14-2; Clavispora lusitaniae, PD 31-1; D. hansenii, PD 33-1; D. hansenii, PD 43-1; D. hansenii, PD 13-1; D. hansenii, PD 49-1; D. hansenii, PD 12-1; D. hansenii, PD 2-1; D. hansenii, PD 32-1; D. hansenii, PD 49-2; D. hansenii, PD 19-1; D. hansenii, PD 12-2; D. hansenii, PD 29-1; D. hansenii, PD 25-2; D. hansenii, PD 34-1; T. delbrueckii, PD 39-1; T. delbrueckii, PD 47-1; T. delbrueckii, PD 56-2; T. delbrueckii, PD 36-1; K. lactis, PD 36-2; K. lactis).

3.3 Antagonistic effect of the yeast strains

Only three of the yeast strains isolated from Kashkaval cheese samples (11-1, 49-1, 49-2) showed antagonistic effect (halos between 8.0-12.0 mm) against Listeria monocytogenes ATCC 7644. These strains were identified as Rhodotorula mucilaginosa (11-1) and D. hansenii (49-1, 49-2). The most effective antilisterial activity (12 mm) was determined in the Rhodotorula mucilaginosa (11-1) isolates (Table 1). The antagonistic effect against other tested pathogen bacteria was not shown .

3.4 Proteolytic and lipolytic activities of the yeast strains

Only the strain 36-1, identified as K. lactis and isolated from an Orgu cheese sample (Table 1), showed proteolytic activity. Lipolytic activity was not observed on tributyrin agar.

4 Discussion

Yeast counts are determined at a high level in cheeses (e.g., 106-109 cfu/g), even in cheeses produced by bacterial starters (Capece & Romano, 2009Capece, A., & Romano, P. (2009). “Pecorino di Filiano” cheese as a selective habitat for the yeast species, Debaryomyces hansenii, Short communication. International Journal of Food Microbiology, 132(2-3), 180-184. http://dx.doi.org/10.1016/j.ijfoodmicro.2009.04.007. PMid:19411124.
http://dx.doi.org/10.1016/j.ijfoodmicro....
; Karasu-Yalcin et al., 2017Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2017). Enzymatic characterization of yeast strains originated from traditional Mihalic cheese. Journal of Microbiology, Biotechnology and Food Sciences, 6(5), 1152-1156. http://dx.doi.org/10.15414/jmbfs.2017.6.5.1152-1156.
http://dx.doi.org/10.15414/jmbfs.2017.6....
). In the present study, the yeast loads were found as too low (1.47-4.71 log cfu g-1) to affect in cheese ripening. The regulation limiting yeast and mold counts (1-2 log cfu g-1) (Turkish, 2010Tarım ve Köyişleri Bakanlığı. (2010). Türk Gida Kodeksi Mikrobiyolojik Kriterler Tebliği (No. 27456). Resmî Gazete’de yayımlanarak yürürlüğe giren Türk Gıda Kodeksi. Retrieved from http://www.resmigazete.gov.tr/eskiler/2010/01/20100108-10.htm
http://www.resmigazete.gov.tr/eskiler/20...
) and limited artisanal cheese production are thought to be the reasons of the low yeast counts in this study. The yeast count of Kashkaval cheese, a kind of pasta filata cheese, was determined in this study to range between 1.47 and 3.81 log cfu g-1. Çetinkaya & Soyutemiz (2006)Çetinkaya, F., & Soyutemiz, G. E. (2006). Microbiological and chemical changes throughout the manufacture and ripening of Kashar: a traditional Turkish cheese. Turkish Journal of Veterinary and Animal Sciences, 30, 397-404. reported the yeast and mold counts of Kashkaval cheese between 3.41 and 3.91 log cfu g-1. There are some factors during the processing of Kashkaval cheese for reducing yeast count like heat treatment (75 ± 1 ºC for 5 min) of the curd during the stretching process (Çetinkaya & Soyutemiz, 2006Çetinkaya, F., & Soyutemiz, G. E. (2006). Microbiological and chemical changes throughout the manufacture and ripening of Kashar: a traditional Turkish cheese. Turkish Journal of Veterinary and Animal Sciences, 30, 397-404.), and some chemical preservatives, which are allowed in limited amounts such as sorbates and benzoates, may also reduce yeast count (Ozdemir & Demirci, 2006Ozdemir, C., & Demirci, M. (2006). Selected microbiological properties of Kashar cheese samples preserved with potassium sorbate. International Journal of Food Properties, 9(3), 515-521. http://dx.doi.org/10.1080/10942910600596191.
http://dx.doi.org/10.1080/10942910600596...
).

Yeasts are widely isolated from cheeses because of ability to live at acidic pH, high salted environments, and storage at low temperatures of these foods. Yeasts may cause spoilage on dairy products contaminated through raw materials, additives, and other ingredients, and that may also occur due to environmental contamination, including surfaces of equipment (Fleet, 1990Fleet, G. H. (1990). Yeasts in dairy products. The Journal of Applied Bacteriology, 68(3), 199-211. http://dx.doi.org/10.1111/j.1365-2672.1990.tb02566.x. PMid:2187843.
http://dx.doi.org/10.1111/j.1365-2672.19...
; Jakobsen & Narvhus, 1996Jakobsen, M., & Narvhus, J. (1996). Yeasts and their possible beneficial and negative effects on the quality of dairy products. International Dairy Journal, 6(8-9), 755-768. http://dx.doi.org/10.1016/0958-6946(95)00071-2.
http://dx.doi.org/10.1016/0958-6946(95)0...
). However, some yeast strains have a desirable flavor and taste development during the ripening stage (Fleet, 1990Fleet, G. H. (1990). Yeasts in dairy products. The Journal of Applied Bacteriology, 68(3), 199-211. http://dx.doi.org/10.1111/j.1365-2672.1990.tb02566.x. PMid:2187843.
http://dx.doi.org/10.1111/j.1365-2672.19...
).

In this study, the predominant isolated species from cheese samples was D. hansenii, especially in Tulum and Mihalic cheese. T. delbrueckii, K. lactis, C. parapsilosis, Cl. lusitaniae, S. cerevisiae, K. marxianus, R. mucilaginosa, Meyerozyma guilliermondii, C. zeylanoides, and C. albicans were found as other identified yeast species. It was mentioned that yeast species belonging to Debaryomyces, Candida, Kluyveromyces, Trichosporon, Saccharomyces, Pichia, Zygosaccharomyces and Geotrichum genera were the most generally isolated species from Turkish white pickled, Tulum and Kashkaval cheeses (Hayaloglu et al., 2002Hayaloglu, A. A., Guven, M., & Fox, P. F. (2002). Microbiological, biochemical and technological properties of Turkish white cheese ‘Beyaz Peynir’ Review. International Dairy Journal, 12(8), 635-648. http://dx.doi.org/10.1016/S0958-6946(02)00055-9.
http://dx.doi.org/10.1016/S0958-6946(02)...
; Kavas et al., 2006Kavas, G., Kinik, O., Uysal, S., Kilic, S., Celikel, N., & Akbulut, N. (2006). Characterisation of yeasts isolated from artisanal Turkish dairy products. International Journal of Dairy Science, 1, 44-50. http://dx.doi.org/10.3923/ijds.2006.44.50.
http://dx.doi.org/10.3923/ijds.2006.44.5...
; Karasu-Yalcin et al., 2012Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2012). Identification and enzymatic characterization of the yeasts isolated from Erzincan Tulum cheese. Mljekarstvo, 62, 53-61.). Candida, Geotrichum, and Trichosporon genera were also identified as dominant yeast flora for Mihalic cheese (Karasu-Yalcin et al., 2017Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2017). Enzymatic characterization of yeast strains originated from traditional Mihalic cheese. Journal of Microbiology, Biotechnology and Food Sciences, 6(5), 1152-1156. http://dx.doi.org/10.15414/jmbfs.2017.6.5.1152-1156.
http://dx.doi.org/10.15414/jmbfs.2017.6....
). It was mentioned that D. hansenii was the most widely found species in almost all cheeses types (Fleet, 1990Fleet, G. H. (1990). Yeasts in dairy products. The Journal of Applied Bacteriology, 68(3), 199-211. http://dx.doi.org/10.1111/j.1365-2672.1990.tb02566.x. PMid:2187843.
http://dx.doi.org/10.1111/j.1365-2672.19...
; Capece & Romano, 2009Capece, A., & Romano, P. (2009). “Pecorino di Filiano” cheese as a selective habitat for the yeast species, Debaryomyces hansenii, Short communication. International Journal of Food Microbiology, 132(2-3), 180-184. http://dx.doi.org/10.1016/j.ijfoodmicro.2009.04.007. PMid:19411124.
http://dx.doi.org/10.1016/j.ijfoodmicro....
; Karasu-Yalcin et al., 2017Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2017). Enzymatic characterization of yeast strains originated from traditional Mihalic cheese. Journal of Microbiology, Biotechnology and Food Sciences, 6(5), 1152-1156. http://dx.doi.org/10.15414/jmbfs.2017.6.5.1152-1156.
http://dx.doi.org/10.15414/jmbfs.2017.6....
). Debaryomyces hansenii has excellent tolerance to salty environments and proteolytic and lipolytic enzyme activty that are important for cheese ripening (Jakobsen & Narvhus, 1996Jakobsen, M., & Narvhus, J. (1996). Yeasts and their possible beneficial and negative effects on the quality of dairy products. International Dairy Journal, 6(8-9), 755-768. http://dx.doi.org/10.1016/0958-6946(95)00071-2.
http://dx.doi.org/10.1016/0958-6946(95)0...
; Capece & Romano, 2009Capece, A., & Romano, P. (2009). “Pecorino di Filiano” cheese as a selective habitat for the yeast species, Debaryomyces hansenii, Short communication. International Journal of Food Microbiology, 132(2-3), 180-184. http://dx.doi.org/10.1016/j.ijfoodmicro.2009.04.007. PMid:19411124.
http://dx.doi.org/10.1016/j.ijfoodmicro....
). Karasu-Yalcin et al. (2017)Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2017). Enzymatic characterization of yeast strains originated from traditional Mihalic cheese. Journal of Microbiology, Biotechnology and Food Sciences, 6(5), 1152-1156. http://dx.doi.org/10.15414/jmbfs.2017.6.5.1152-1156.
http://dx.doi.org/10.15414/jmbfs.2017.6....
emphasized that it was expected to find D. hansenii as dominant yeast microflora of Mihalic cheese, a salty traditional cheese in Turkey.

Some yeast strains may show an antagonistic effect against pathogen bacteria by producing antibacterial compounds called mycocins (killer toxins). These compounds are glycoproteins or extracellular proteins which damage the cell membrane function in the targeted microorganisms (Hatoum et al., 2013Hatoum, R., Labrie, S., & Fliss, I. (2013). Identification and partial characterization of antilisterial compounds produced by dairy yeasts. Probiotics and Antimicrobial Proteins, 5(1), 8-17. http://dx.doi.org/10.1007/s12602-012-9109-8. PMid:26782600.
http://dx.doi.org/10.1007/s12602-012-910...
). Although the antimicrobial effects of yeasts against other yeasts by killer toxins are well analyzed (Goerges et al., 2006Goerges, S., Aigner, U., Silakowski, B., & Scherer, S. (2006). Inhibition of Listeria monocytogenes by food-borne yeasts. Applied and Environmental Microbiology, 72(1), 313-331. http://dx.doi.org/10.1128/AEM.72.1.313-318.2006. PMid:16391059.
http://dx.doi.org/10.1128/AEM.72.1.313-3...
; Buyuksirit & Kuleasan, 2014Buyuksirit, T., & Kuleasan, H. (2014). Antimicrobial agents produced by yeasts. World Academy of Science, Engineering and Technology International Journal of Biological, Veterinary, Agricultural, and Food Engineering, 8, 999-1002.), studies conducted to find out the antibacterial effects of yeasts against Listeria monocytogenes and Staphylococcus aureus are limited (Goerges et al., 2006Goerges, S., Aigner, U., Silakowski, B., & Scherer, S. (2006). Inhibition of Listeria monocytogenes by food-borne yeasts. Applied and Environmental Microbiology, 72(1), 313-331. http://dx.doi.org/10.1128/AEM.72.1.313-318.2006. PMid:16391059.
http://dx.doi.org/10.1128/AEM.72.1.313-3...
; Roostita et al., 2011Roostita, L. B., Fleet, G. H., Wendry, S. P., Apon, Z. M., & Gemilang, L. U. (2011). Determination of yeasts antimicrobial activity in milk and meat products. Advance Journal of Food Science and Technology, 3, 442-445.; Hatoum et al., 2013Hatoum, R., Labrie, S., & Fliss, I. (2013). Identification and partial characterization of antilisterial compounds produced by dairy yeasts. Probiotics and Antimicrobial Proteins, 5(1), 8-17. http://dx.doi.org/10.1007/s12602-012-9109-8. PMid:26782600.
http://dx.doi.org/10.1007/s12602-012-910...
). Goerges et al. (2006)Goerges, S., Aigner, U., Silakowski, B., & Scherer, S. (2006). Inhibition of Listeria monocytogenes by food-borne yeasts. Applied and Environmental Microbiology, 72(1), 313-331. http://dx.doi.org/10.1128/AEM.72.1.313-318.2006. PMid:16391059.
http://dx.doi.org/10.1128/AEM.72.1.313-3...
reported that C. intermedia, K. marxianus, and Pichia norvegensis yeast strains isolated from cheeses showed strong antilisterial activity. Roostita et al. (2011)Roostita, L. B., Fleet, G. H., Wendry, S. P., Apon, Z. M., & Gemilang, L. U. (2011). Determination of yeasts antimicrobial activity in milk and meat products. Advance Journal of Food Science and Technology, 3, 442-445., reported that a C. parapsilosis isolate from yoghurt with fruits showed antibacterial activity against Pseudomonas aeruginosa, Escherichia coli and S. aureus. It has been reported that Candida catenulata, C. parapsilosis, C. tropicalis, D. hansenii, G. candidum, P. fermentans, and P. anomala strains obtained from raw milk and cheese products showed antilisterial activity against Listeria ivanovii. In this study, one R. mucilaginosa (11-1) and two D. hansenii (49-1, 49-2) strains exhibited antibacterial activity against L. monocytogenes. These three strains were isolated from Kashkaval cheese samples. Although a study about antilisterial activity potential of D. hansenii (Hatoum et al., 2013Hatoum, R., Labrie, S., & Fliss, I. (2013). Identification and partial characterization of antilisterial compounds produced by dairy yeasts. Probiotics and Antimicrobial Proteins, 5(1), 8-17. http://dx.doi.org/10.1007/s12602-012-9109-8. PMid:26782600.
http://dx.doi.org/10.1007/s12602-012-910...
) has been published, to our knowledge, this research is the first report describing the antimicrobial activity of Rhodotorula mucilaginosa against Listeria monocytogenes. R. mucilaginosa has commonly been found in the microbial flora of some foods and beverages such as cherries, peanuts, fresh fruits, apple cider, fruit juice, sausages, cheese, crustaceans and edible mollusks, and has also been found in air, seawater, freshwater, and goat’s milk (Wirth & Goldani, 2012Wirth, F., & Goldani, L. Z. (2012). Epidemiology of Rhodotorula: an emerging pathogen. Interdisciplinary Perspectives on Infectious Diseases, 465717. https://doi.org/10.1155/2012/465717.
https://doi.org/10.1155/2012/465717...
).

As a result of screening the proteolytic and lipolytic activity potential of the yeast isolates, proteolytic activity was only found in one K. lactis from Orgu cheese, and none of the isolates showed lipolytic activity in this study. Flores et al. (1999)Flores, M. F., Cuellas, A., & Voget, C. E. (1999). The proteolytic system of the yeast Kluyveromyces lactis. Yeast, 15(14), 1437-1448. http://dx.doi.org/10.1002/(SICI)1097-0061(199910)15:14<1437::AID-YEA445>3.0.CO;2-C. PMid:10514562.
http://dx.doi.org/10.1002/(SICI)1097-006...
also described proteolytic Kluyveromyces lactis strains.

5 Conclusion

Debaryomyces hansenii and Torulaspora delbrueckii were identified as predominant yeast species in traditional Turkish cheese samples. Although antilisterial and proteolytic activity potentials were determined in a limited number of strains, there is a need for more extensive studies to determine the technological properties of the yeast strains from natural yeast microflora of different traditional Turkish cheeses with a view to its possible use in the dairy industry.

  • Practical Application: The wild yeast microflora from some traditional Turkish cheeses were identified by means of molecular techniques. These information may be useful for quality and shelf life of Turkish cheeses.

References

  • Aday, S., & Karagul Yuceer, Y. (2014). Physicochemical and sensory properties of Mihalic cheese. International Journal of Food Properties, 17(10), 2207-2227. http://dx.doi.org/10.1080/10942912.2013.790904
    » http://dx.doi.org/10.1080/10942912.2013.790904
  • Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W., & Lipman, D. J. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research, 25(17), 3389-3402. http://dx.doi.org/10.1093/nar/25.17.3389 PMid:9254694.
    » http://dx.doi.org/10.1093/nar/25.17.3389
  • Banjara, N., Suhr, M. J., & Hallen-Adams, H. E. (2015). Diversity of yeast and mold species from a variety of cheese types. Current Microbiology, 70(6), 792-800. http://dx.doi.org/10.1007/s00284-015-0790-1 PMid:25694357.
    » http://dx.doi.org/10.1007/s00284-015-0790-1
  • Büchl, N. R., & Seiler, H. (2011). Yeast and molds: yeasts in milk and dairy products. In H. Roginski, J. W. Fuquay, P. F. Fox, eds. Encyclopedia of dairy sciences (2nd ed., pp. 744-753). New York: Elsevier. http://dx.doi.org/10.1016/B978-0-12-374407-4.00498-2
    » http://dx.doi.org/10.1016/B978-0-12-374407-4.00498-2
  • Buyuksirit, T., & Kuleasan, H. (2014). Antimicrobial agents produced by yeasts. World Academy of Science, Engineering and Technology International Journal of Biological, Veterinary, Agricultural, and Food Engineering, 8, 999-1002.
  • Capece, A., & Romano, P. (2009). “Pecorino di Filiano” cheese as a selective habitat for the yeast species, Debaryomyces hansenii, Short communication. International Journal of Food Microbiology, 132(2-3), 180-184. http://dx.doi.org/10.1016/j.ijfoodmicro.2009.04.007 PMid:19411124.
    » http://dx.doi.org/10.1016/j.ijfoodmicro.2009.04.007
  • Capece, A., Pietrafesa, R., & Romano, P. (2011). Experimental approach for target selection of wild wine yeasts from spontaneous fermentation of “Inzolia” grapes. World Journal of Microbiology & Biotechnology, 27(12), 2775-2783. http://dx.doi.org/10.1007/s11274-011-0753-z
    » http://dx.doi.org/10.1007/s11274-011-0753-z
  • Çetinkaya, F., & Soyutemiz, G. E. (2006). Microbiological and chemical changes throughout the manufacture and ripening of Kashar: a traditional Turkish cheese. Turkish Journal of Veterinary and Animal Sciences, 30, 397-404.
  • Corbaci, C., Ucar, F. B., & Yalcin, H. T. (2012). Isolation and characterization of yeasts associated with Turkish-style homemade dairy products and their potential as starter cultures. African Journal of Microbiological Research, 6, 534-542.
  • Esteve-Zarzoso, B., Belloch, C., Uruburu, F., & Querol, A. (1999). Identification of yeasts by RFLP analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers. International Journal of Systematic Bacteriology, 49(Pt 1), 329-337. PMid:10028278.
  • Ferreira, A. D., & Viljoen, B. C. (2003). Yeasts as adjunct starters in matured Cheddar cheese. International Journal of Food Microbiology, 86(1-2), 131-140. http://dx.doi.org/10.1016/S0168-1605(03)00252-6 PMid:12892928.
    » http://dx.doi.org/10.1016/S0168-1605(03)00252-6
  • Fleet, G. H. (1990). Yeasts in dairy products. The Journal of Applied Bacteriology, 68(3), 199-211. http://dx.doi.org/10.1111/j.1365-2672.1990.tb02566.x PMid:2187843.
    » http://dx.doi.org/10.1111/j.1365-2672.1990.tb02566.x
  • Flores, M. F., Cuellas, A., & Voget, C. E. (1999). The proteolytic system of the yeast Kluyveromyces lactis. Yeast, 15(14), 1437-1448. http://dx.doi.org/10.1002/(SICI)1097-0061(199910)15:14<1437::AID-YEA445>3.0.CO;2-C PMid:10514562.
    » http://dx.doi.org/10.1002/(SICI)1097-0061(199910)15:14<1437::AID-YEA445>3.0.CO;2-C
  • Goerges, S., Aigner, U., Silakowski, B., & Scherer, S. (2006). Inhibition of Listeria monocytogenes by food-borne yeasts. Applied and Environmental Microbiology, 72(1), 313-331. http://dx.doi.org/10.1128/AEM.72.1.313-318.2006 PMid:16391059.
    » http://dx.doi.org/10.1128/AEM.72.1.313-318.2006
  • Hansen, T. K., & Jakobsen, M. (2001). Taxonomical and technological characteristics of Saccharomyces spp. associated with blue-veined cheese. International Journal of Food Microbiology, 69(1-2), 59-68. http://dx.doi.org/10.1016/S0168-1605(01)00573-6 PMid:11589561.
    » http://dx.doi.org/10.1016/S0168-1605(01)00573-6
  • Harris, L. J., Daeschel, M. A., Stiles, M. E., & Klaenhammer, T. R. (1989). Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes. Journal of Food Protection, 52(6), 384-387. http://dx.doi.org/10.4315/0362-028X-52.6.384 PMid:31003307.
    » http://dx.doi.org/10.4315/0362-028X-52.6.384
  • Hatoum, R., Labrie, S., & Fliss, I. (2012). Antimicrobial and probiotic properties of yeasts: from fundamental to novel applications. Frontiers in Microbiology, 3 (421). http://dx.doi.org/10.3389/fmicb.2012.00421
    » http://dx.doi.org/10.3389/fmicb.2012.00421
  • Hatoum, R., Labrie, S., & Fliss, I. (2013). Identification and partial characterization of antilisterial compounds produced by dairy yeasts. Probiotics and Antimicrobial Proteins, 5(1), 8-17. http://dx.doi.org/10.1007/s12602-012-9109-8 PMid:26782600.
    » http://dx.doi.org/10.1007/s12602-012-9109-8
  • Hayaloglu, A. A., Guven, M., & Fox, P. F. (2002). Microbiological, biochemical and technological properties of Turkish white cheese ‘Beyaz Peynir’ Review. International Dairy Journal, 12(8), 635-648. http://dx.doi.org/10.1016/S0958-6946(02)00055-9
    » http://dx.doi.org/10.1016/S0958-6946(02)00055-9
  • Jakobsen, M., & Narvhus, J. (1996). Yeasts and their possible beneficial and negative effects on the quality of dairy products. International Dairy Journal, 6(8-9), 755-768. http://dx.doi.org/10.1016/0958-6946(95)00071-2
    » http://dx.doi.org/10.1016/0958-6946(95)00071-2
  • Kamber, U. (2008). The traditional cheeses of Turkey: cheeses common to all regions. Food Reviews International, 24(1), 1-38. http://dx.doi.org/10.1080/87559120701761833
    » http://dx.doi.org/10.1080/87559120701761833
  • Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2012). Identification and enzymatic characterization of the yeasts isolated from Erzincan Tulum cheese. Mljekarstvo, 62, 53-61.
  • Karasu-Yalcin, S., Senses-Ergul, S., & Ozbas, Z. Y. (2017). Enzymatic characterization of yeast strains originated from traditional Mihalic cheese. Journal of Microbiology, Biotechnology and Food Sciences, 6(5), 1152-1156. http://dx.doi.org/10.15414/jmbfs.2017.6.5.1152-1156
    » http://dx.doi.org/10.15414/jmbfs.2017.6.5.1152-1156
  • Kavas, G., Kinik, O., Uysal, S., Kilic, S., Celikel, N., & Akbulut, N. (2006). Characterisation of yeasts isolated from artisanal Turkish dairy products. International Journal of Dairy Science, 1, 44-50. http://dx.doi.org/10.3923/ijds.2006.44.50
    » http://dx.doi.org/10.3923/ijds.2006.44.50
  • Lanciotti, R., Chaves-López, C., Patrignani, F., Paparella, A., Guerzoni, M. E., Serio, A., & Suzzi, G. (2004). Effects of milk treatment with dynamic high pressure on microbial populations, and lipolytic and proteolytic profiles of Crescenza cheese. International Journal of Dairy Technology, 57(1), 19-25. http://dx.doi.org/10.1111/j.1471-0307.2004.00121.x
    » http://dx.doi.org/10.1111/j.1471-0307.2004.00121.x
  • National Center for Biotechnology Information – NCBI. (2015). Bethesda: NCBI. Retrieved from http://www.ncbi.nlm.nih.gov
    » http://www.ncbi.nlm.nih.gov
  • Ozdemir, C., & Demirci, M. (2006). Selected microbiological properties of Kashar cheese samples preserved with potassium sorbate. International Journal of Food Properties, 9(3), 515-521. http://dx.doi.org/10.1080/10942910600596191
    » http://dx.doi.org/10.1080/10942910600596191
  • Roostita, L. B., Fleet, G. H., Wendry, S. P., Apon, Z. M., & Gemilang, L. U. (2011). Determination of yeasts antimicrobial activity in milk and meat products. Advance Journal of Food Science and Technology, 3, 442-445.
  • Tarım ve Köyişleri Bakanlığı. (2010). Türk Gida Kodeksi Mikrobiyolojik Kriterler Tebliği (No. 27456). Resmî Gazete’de yayımlanarak yürürlüğe giren Türk Gıda Kodeksi Retrieved from http://www.resmigazete.gov.tr/eskiler/2010/01/20100108-10.htm
    » http://www.resmigazete.gov.tr/eskiler/2010/01/20100108-10.htm
  • Vasdinyei, R., & Deak, T. (2003). Characterization of yeast isolates originating from Hungarian dairy products using traditional and molecular identification techniques. International Journal of Food Microbiology, 86(1-2), 123-130. http://dx.doi.org/10.1016/S0168-1605(03)00251-4 PMid:12892927.
    » http://dx.doi.org/10.1016/S0168-1605(03)00251-4
  • Wirth, F., & Goldani, L. Z. (2012). Epidemiology of Rhodotorula: an emerging pathogen. Interdisciplinary Perspectives on Infectious Diseases, 465717. https://doi.org/10.1155/2012/465717
    » https://doi.org/10.1155/2012/465717

Publication Dates

  • Publication in this collection
    08 May 2020
  • Date of issue
    Oct-Dec 2020

History

  • Received
    03 Sept 2019
  • Accepted
    04 Nov 2019
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