PCR verification of microplate phenotypic system identification for LAB from traditional Western Balkan raw milk cheeses

Abstract

Fermentation and ripening specificity of traditional cheeses are predominantly directed by the natural microbial community present in milk selected by the cheese-making environment and technology. Therefore the traditional cheeses are unique products with specific microbiota biodiversity. There are several approaches for the identification of microbial population, however all of them have certain advantages and disadvantages. In this study the eligibility and performance of the Biolog phenotypic identification system (Biolog, Inc.) with GEN III microplates was tested. Parallel to this method, polymerase chain reaction with genus-and species-specific primers was performed. One hundred sixty-five isolates from nine types of artisan cheeses were isolated and analysed. Cheeses were produced from raw ewe's milk in Slovenia, Bosnia and Herzegovina, Croatia and Serbia. The Biolog phenotypic identification system identified 90 isolates, but only 55 identifications acquired by the Biolog system were supported by polymerase chain reaction at a genus level and 28 at a species level. The obtained results showed that the reliability of commercial phenotypic identification systems was inadequate when analysing lactic acid bacteria isolates from natural, spontaneous fermentations and needs to be additionally corroborated by genotypic identification methods.

Diversity of non-starter lactic acid bacteria in autochthonous dairy products from Western Balkan Countries - Technological and probiotic properties

“…White brined cheeses are full-fat cheeses of intense flavor, dry-salted, ripened and stored in brine. They are usually consumed after several months of ripening and have a particular flavor depending on the milk used (cow's, ewe's and goat's milk or a mixture of two types of milk) and their microbiota varies depending on the ripening period (Begovic et al, 2011;Levkov, Srbinovska, & Gjorgovska, 2014;Levkov, Mojsova, Nastova, Srbinovska, & Gjorgovska, 2017;Mojsova et al, 2013;Nikolic et al, 2008;Paveljšek, Trmčić, Hacin, & Rogelj, 2014;Pešić-Mikulec & Jovanović, 2005;Terzic-Vidojevic, Vukasinovic, Veljovic, Ostojic, & Topisirovic, 2007;Terzic-Vidojevic et al, 2009a;Terzic-Vidojevic, Lozo, & Topisirovic, 2009b;Terzic-Vidojevic et al, 2013, 2014a, 2015b. The ripening period also affects the specific texture and flavor of the final product (Asteri et al, 2009;Callon, Berdagué, Dufour, & Montel, 2005;Farahani, Rasooli, & Owlia, 2017;Pino et al, 2019;Saiki et al, 2018).…”

“…Hard cheeses are another group of dairy products made in the countries of the Western Balkans (Bojanic Rasovic, Mayrhofer, Martinovic, Dürr, & Doming, 2017;Čanžek Majhenič, Rogelj, & Perko, 2005;Čanžek Majhenič, Mohar Lorberg, & Rogelj, 2007;Levkov & Kakurinov, 2012;Levkov, Srbinovska, & Gjorgovska, 2014;Levkov, Mojsova, Nastova, Srbinovska, & Gjorgovska, 2017;Mrkonjić Fuka, Engel, Skelin, Redžepović, & Schloter, 2010;Mrkonjić Fuka et al, 2013;Paveljšek, Trmčić, Hacin, & Rogelj, 2014;Skelin et al, 2012). The main characteristics of hard cheeses are their very low moisture content and longer shelf life.…”

In vitro evaluation of the antimicrobial potential of Streptococcus uberis isolated from a local cheese from Southeastern Serbia

“…traditional agricultural products, especially cheeses, are recognizable by their authentic flavour and texture. The nutrition of dairy cows, the specific attributes of raw milk as well as the basic traditional cheese making process and the natural bacteria responsible for the fermentation process and ripening, have a very significant role in the formation of the flavour and organoleptic properties of raw milk cheeses (PRPIĆ et al, 2003;PaVeLJŠek et al, 2014;MIreCkI et al, 2015).…”