Determination of cheese origin by using 16S rDNA fingerprinting of bacteria communities by PCR–DGGE: Preliminary application to traditional Minas cheese

Abstract The bacterial diversity occurring in traditional Minas cheese, made from raw milk, produced in four different regions of Minas Gerais state in Brazil, namely Cerrado, Araxa, Serro and Serra da Canastra, were evaluated by PCR–DGGE analysis of the V3 region of the bacterial 16S rDNA using primers previously described in the literature. Samples of Minas cheese made with pasteurized milk from Serro region were used as control. Bacterial DNA templates for PCR were directly extracted from cheese samples in a unique step. The PCR–DGGE revealed 1 band for the cheese samples made from pasteurized milk and 7 to 13 bands for those from traditional cheeses. Some bands were common for all cheeses and some were unique for each region. The results suggest that specific band profiles from traditional cheeses from each region may be used as a biological bar code to discriminate their origins. In addition, the band profiles clearly distinguished cheese made from pasteurized and raw milk. DGGE band sequencing analysis showed that species belonging to the genera Streptococcus may predominate in the traditional Minas cheese.

[1]  J. Guyot,et al.  Polyphasic Study of the Spatial Distribution of Microorganisms in Mexican Pozol, a Fermented Maize Dough, Demonstrates the Need for Cultivation-Independent Methods To Investigate Traditional Fermentations , 1999, Applied and Environmental Microbiology.

[2]  V. Nam,et al.  Application of PCR-DGGE in determining food origin: Cases studies of fish and fruits , 2008 .

[3]  D. Montēt,et al.  Determination of fish origin by using 16S rDNA fingerprinting of bacterial communities by PCR-DGGE : An application on Pangasius fish from Viet Nam , 2008 .

[4]  Danilo Ercolini,et al.  PCR-DGGE fingerprinting: novel strategies for detection of microbes in food. , 2004, Journal of microbiological methods.

[5]  L. Cocolin,et al.  Microbial diversity, dynamics and activity throughout manufacturing and ripening of Castelmagno PDO cheese. , 2010, International journal of food microbiology.

[6]  Didier Montet,et al.  Determination of fruit origin by using 26S rDNA fingerprinting of yeast communities by PCR–DGGE: preliminary application to Physalis fruits from Egypt , 2009, Yeast.

[7]  C. Mbofung,et al.  Study of the microbial diversity of Oreochromis niloticus of three lakes of Cameroon by PCR-DGGE: Application to the determination of the geographical origin , 2010 .

[8]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[9]  K. Baiyeri,et al.  Fruit characteristics and ripening pattern of ten Musa genotypes in a sub-humid environment in Nigeria , 2008 .

[10]  C. A. Rosa,et al.  Bactérias do acido láctico e leveduras associadas com o queijo-de-minas artesanal produzido na região da Serra do Salitre, Minas Gerais , 2009 .

[11]  D. Ercolini,et al.  Microbial diversity in natural whey cultures used for the production of Caciocavallo Silano PDO cheese. , 2008, International journal of food microbiology.

[12]  D. Ercolini,et al.  Relationships between flavoring capabilities, bacterial composition, and geographical origin of natural whey cultures used for traditional water-buffalo mozzarella cheese manufacture. , 2003, Journal of dairy science.

[13]  D. Ercolini,et al.  PCR–DGGE fingerprints of microbial succession during a manufacture of traditional water buffalo mozzarella cheese , 2004, Journal of applied microbiology.

[14]  H. Kim,et al.  Analysis of Bacterial and Fungal Communities in Japanese- and Chinese-Fermented Soybean Pastes Using Nested PCR–DGGE , 2010, Current Microbiology.

[15]  P. Vos,et al.  Applicability of combined amplified ribosomal DNA restriction analysis (ARDRA) patterns in bacterial phylogeny and taxonomy , 1996 .

[16]  C. Mbofung,et al.  Comparison of bacterial communities of tilapia fish from Cameroon and Vietnam using PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) , 2009 .

[17]  D. Montēt,et al.  Determination of citrus fruit origin by using 16S rDNA fingerprinting of bacterial communities by PCR- DGGE: an application to clementine from Morocco and Spain , 2008 .

[18]  B. Thomma,et al.  Recent advances in molecular techniques to study microbial communities in food-associated matrices and processes. , 2008, Food microbiology.

[19]  V. Sheffield,et al.  Attachment of a 40-base-pair G + C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[20]  G. Franco,et al.  Yeast populations associated with the artisanal cheese produced in the region of Serra da Canastra, Brazil , 2006 .

[21]  D. Ercolini,et al.  Molecular evaluation of microbial diversity occurring in different types of Mozzarella cheese , 2001, Journal of applied microbiology.