Evaluation of lactic acid bacteria and yeast diversity in traditional white pickled and fresh soft cheeses from the mountain regions of Serbia and lowland regions of Croatia
Introduction
The Western Balkan Countries (WBC) constitute a distinct geographical region in Europe with long experience in the production of traditional dairy foods by spontaneous or controlled fermentation of cow's, ewe's and goat's milk. In this region, various spontaneously fermented dairy products, such as white pickled and fresh soft cheeses, are manufactured in households without the use of commercial starter cultures. Most of these products are made from unpasteurised milk and the composition of the “natural starter” depends solely on the microbiota present in the raw milk or in the local environment. These microorganisms contribute to preservation, flavour, aroma and texture, determining unique product characteristics.
Preliminary characterisation of the dominant microbiota in these artisanal dairy products revealed that they contain a considerable diversity of lactic acid bacteria (LAB) and yeasts. LAB isolated from autochthonous cheeses have considerable genetic, metabolic and technological potential: good milk protein coagulation, accelerated acidification, production of proteinases, exopolysaccharides, aroma and taste precursors, as well as health-promoting properties (Topisirovic et al., 2006). The dominant LAB are represented by various lactobacillus species: leuconostocs, enterococci, Streptococcus thermophilus and Lactococcus lactis (Randazzo et al., 2002, Terzic-Vidojevic et al., 2007, Nikolic et al., 2008, Jokovic et al., 2008, Chebeňová-Turcovská et al., 2011). Yeast species usually represent secondary microbiota in dairy products and among them Kluyveromyces marxianus, Debaryomyces hansenii and Saccharomyces cerevisiae are usually found to be predominant (Frolich-Wyder, 2003, Chebeňová-Turcovská et al., 2011). Moreover, recent studies have indicated that food borne yeasts D. hansenii and Yarrowia lipolytica could successfully be used as part of starter cultures for cheese manufacturing, enhancing flavour development during cheese maturation (Ferreira and Viljoen, 2003, Sørensen et al., 2011).
Considering the significant role of autochthonous microbiota in the determination of the main features of a cheese, the goal of this study was the identification and characterisation of LAB and yeasts associated with the production of artisanal white pickled and fresh soft cheeses traditionally performed in rural households in the mountain regions of Serbia and lowland regions of Croatia. To our knowledge, this is the most complete study highlighting the phenotypic and genotypic diversity of LAB and natural yeast isolates from traditional cheeses in Serbia and Croatia.
Section snippets
Cheese samples
The cheese sampling regions were chosen based on the history of traditional dairy product manufacturing from different kinds of raw milk without the addition of any starter culture. Nine white pickled artisanal cheeses (designated as BG) were collected from three regions in Serbia: the South Morava region (one-day-old BGAL2, two-day-old BGLE1, and 10-day-old BGAL3), Golija Mountain (10-day-old BGGO5 and BGGO11, and 60-day-old BGGO7) and the mountainous region of Eastern Serbia (three-day-old
Enumeration, isolation and identification of lactic acid bacteria from cheese samples
Eighteen cheeses from specific localities in Serbia and Croatia were analysed. The viable counts of the main microbial groups present in the cheese samples are shown in Table 1. The total counts of LAB in the nine white pickled (BG) cheese samples, determined on MRS and M17 agar plates, varied in the range of 8.2 log CFU/g (BGGO5) to 9.5 log CFU/g (BGAL3), while in the fresh soft (ZG) cheese samples the total LAB counts varied in the range of 3.3 log CFU/g (ZGPR3) to 10.5 log CFU/g (ZGBP5).
The
Discussion
Strict EU food safety legislation has resulted in lower flexibility in food production and will eventually lead to the disappearance of a number of geographical and artisanal dairy products and their related indigenous microorganisms. The comprehensive evaluation of LAB and yeast diversity obtained in this study highlights the strong influence of unique production processes, such as the use of unpasteurised milk, the absence of starter culture, and the use of homemade rennet and brine, on the
Acknowledgments
This work was funded by the SEE-ERA-NET Plus Project ERA-195/01 (PSALAB), the Ministry of Education and Science of the Republic of Serbia, grant OI 173019, and Ministry of Science, Education and Sports of the Republic of Croatia (Project 0581990 “Probiotics, prebiotics and functional starter cultures”). The authors thank Jure Zupan for his help with figure preparation. Petra Dousová is acknowledged for her technical assistance. We are grateful to Nathaniel Aaron Sprinkle, native English editor
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Permanent address: Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology, Brno, Czech Republic.