Microbiota of Karakačanski skakutanac, an artisanal fresh sheep cheese studied by culture-independent PCR-ARDRA and PCR-DGGE

Abstract

-Karakačanski skakutanac is an artisanal fresh sheep cheese produced on a small scale in a limited area of eastern Croatia. It is manufactured from unrefrigerated raw sheep milk immediately after milking, without the addition of starter culture, and coagulated with industrial rennet. To date, no microbiological or molecular characterization of the biodiversity of the microbiota has been performed. The objective of this study was to obtain an initial insight into the biodiversity of the microbial community associated with this cheese during the production season and shelf life period. Eleven cheeses were obtained from a dairy farm at 14-day intervals during the lactation period of east Friesian sheep in 2007. Bacterial DNA was isolated directly from cheese on the first, second and third day of the cheese shelf life, resulting in a total of 33 DNA samples. Extracted DNA was used as a template for PCR-ARDRA and PCR-DGGE analysis. The use of dual culture-independent approaches revealed similar results and indicated predominance of Lactococcus lactis.

Microbial diversity of the traditional Iranian cheeses Lighvan and Koozeh, as revealed by polyphasic culturing and culture-independent approaches

“…The bacterial and fungal population dynamics recorded by DGGE for the two cheeses were similar to those reported for other traditional cheeses (Flórez and Mayo 2006;Pogacic et al 2010;Randazzo et al 2006). The bacterial diversity of dominant populations was usually greater in milk and curd samples, as some species were never identified in cheese (Table 1).…”

“…The microbiota of fermented foods can now be monitored by a variety of molecular methods (Cocolin et al 2007;JanyandBarbier2008). Approaches combining classical culturing followed by molecular identification of the isolates and application of culture-independent molecular techniques are at present the most reliable choice to obtain objective results about microbial structure and dynamics of any ecosystem (Pogacic et al 2010). The polymerase chain reaction (PCR) coupled with denaturing gradient gel electrophoresis (DGGE)-PCR-DGGE-is one such culture-independent molecular method that has been used to characterise the microbial populations of dairy products (Cocolin et al 2002;Er co li nieta l.2001;Flórez and Mayo 2006;.…”

“…The polymerase chain reaction (PCR) coupled with denaturing gradient gel electrophoresis (DGGE)-PCR-DGGE-is one such culture-independent molecular method that has been used to characterise the microbial populations of dairy products (Cocolin et al 2002;Er co li nieta l.2001;Flórez and Mayo 2006;. In fact, such characterisation has already been performed over the manufacture and ripening of a number of traditional cheeses Ercolini et al 2004;Flórez and Mayo 2006;Pogacic et al 2010;Randazzo et al 2002).…”

Perfil microbiológico del queso de aro consumido en la Cañada Oaxaqueña

“…Galinari et al (2014) obtuvieron un promedio igual al nuestro en quesos artesanales de Brasil. Pogačić et al (2010) mencionan que Lactococcus lactis es la bacteria ácido láctica predominante en queso artesanal elaborado con leche de oveja del este de Croacia. Clavel-Maqueda (2006) reporta un promedio de 6,02 Log 10 UFC/g en muestras de queso Panela, de igual forma menciona que en este tipo de muestras los conteos son muy elevados y podrían afectar la vida de anaquel del producto, provocando un sabor ácido.…”

Probiotic Lactococcus lactis: A Review

“…During the last decades, lots of approaches have been introduced under culture independent molecular methods which are frequently used in the identification and characterization purposes of L. lactis such as Culture-Independent Polymerase Chain Reaction (Salbi et al, 2014;) 16S rDNA Sequencing (Aquilanti et al, 2007;Khemariya et al, 2013b), PCR-Denaturing Gradient Gel Electrophoresis (PDGE) (Mrkonjić Fuka et al, 2010;Pogačić et al, 2010;Marui et al, 2015;), PCR-Temporal Temperature Gradient Electrophoresis (TTGE) (ElBaradei et al, 2008), Single-Strand Conformation Polymorphism-PCR (SSCP-PCR) (Saubusse et al, 2007), Real Time PCR (qPCR) (Grattepanche et al, 2005;Ruggirello et al, 2014), Fluorescence in situ Hybridization (FISH) (Miks-Krajnik and Babuchowski, 2014), Amplified Ribosomal DNA Restriction Analysis (Partial ARDRA) (Delgado and Mayo, 2003), Length Heterogeneity PCR (LH-PCR) (Brusetti et al, 2006), Multilocus Sequence Analysis (MLSA) (Rademarker et al, 2007;Khemariya et al, 2012) (GTG) 5 -PCR Fingerprinting (Rademarker et al, 2007;Khemariya et al, 2014), PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) (Deveau et al, 2003;Khemariya et al, 2013b), Multiple Locus Minisatelite Typing (MLVA) (Que'ne'e et al, 2005), RAPD-PCR (Randomly Amplified Polymorphic DNA-PCR) (Samaržija et al, 2002), Pulse Field Gel Electrophoresis (PFGE) (Campo et al, 2002), Repetitive Element Sequence based PCR (Rep-PCR) (Fernandez et al, 2010), Comparative Genomic Hybridization (CGH) (Taibi et al, 2010), Nested PCR (Khemariya et al, 2013c), Multiplex PCR (Pu et al, 2002) and Sau-PCR (Corich et al, 2005).…”