Characterization of bacterial populations in Danish raw milk cheeses made with different starter cultures by denaturating gradient gel electrophoresis and pyrosequencing

TY - JOUR

T1 - Characterization of bacterial populations in Danish raw milk cheeses made with different starter cultures by denaturating gradient gel electrophoresis and pyrosequencing

AU - Masoud, Wafa Mahmoud Hasan

AU - Takamiya, Monica K Wik

AU - Vogensen, Finn Kvist

AU - Lillevang, Søren

AU - Abu Al-Soud, Waleed

AU - Sørensen, Søren Johannes

AU - Jakobsen, Mogens

PY - 2011/3

Y1 - 2011/3

N2 - The bacterial populations in Danish raw milk cheeses were identified using denaturating gradient gel electrophoresis (DGGE) of PCR amplicons of the V3 region of the 16S rRNA gene and pyrosequencing of tagged amplicons of the V3 and V4 regions of the 16S rRNA gene. Both DNA and RNA extracted from cheeses were studied in order to determine the metabolically active bacteria. The main bacteria, which included Lactococcus, Lactobacillus and Streptococcus, were detected by pyrosequencing and DGGE in both 16S rDNA and cDNA obtained from cheeses indicating their viability and contribution to cheese ripening. Other bacteria like Corynebacterium, Halomonas, Pediococcus, Micrococcus and Staphylococcus, which were encountered in some cheese samples at low percentages compared with the total bacterial populations, were only detected by pyrosequencing. 16S rRNA gene pyrosequencing is an efficient method for deep sequencing of microbial communities and it expands our knowledge of the bacterial diversity in raw milk cheese.

AB - The bacterial populations in Danish raw milk cheeses were identified using denaturating gradient gel electrophoresis (DGGE) of PCR amplicons of the V3 region of the 16S rRNA gene and pyrosequencing of tagged amplicons of the V3 and V4 regions of the 16S rRNA gene. Both DNA and RNA extracted from cheeses were studied in order to determine the metabolically active bacteria. The main bacteria, which included Lactococcus, Lactobacillus and Streptococcus, were detected by pyrosequencing and DGGE in both 16S rDNA and cDNA obtained from cheeses indicating their viability and contribution to cheese ripening. Other bacteria like Corynebacterium, Halomonas, Pediococcus, Micrococcus and Staphylococcus, which were encountered in some cheese samples at low percentages compared with the total bacterial populations, were only detected by pyrosequencing. 16S rRNA gene pyrosequencing is an efficient method for deep sequencing of microbial communities and it expands our knowledge of the bacterial diversity in raw milk cheese.

U2 - 10.1016/j.idairyj.2010.10.007

DO - 10.1016/j.idairyj.2010.10.007

M3 - Journal article

SN - 0958-6946

VL - 21

SP - 142

EP - 148

JO - International Dairy Journal

JF - International Dairy Journal

IS - 3

ER -