Phenotypic and PCR-based characterization of the microflora in Norvegia cheese during ripening

Østlie, Hilde Marit; Eliassen, L.; Florvaag, A.; Skeie, Siv

doi:10.1016/j.ijfoodmicro.2004.01.012

Cited by 47 publications

(30 citation statements)

References 35 publications

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“…The concentration of microorganisms in Cueva de la Magahá cheese was high, ranging from 4.6 to 7.98 log CFU/g, values which correspond to those usually found in cheeses during ripening (4,20,39,42). During the first week the LAB levels were 10 7 CFU/g, and they increased by 1 logarithmic unit during the mid-ripening period to finally stabilize once more at 10 7 CFU/g.…”

Section: Discussionsupporting

confidence: 62%

Polyphasic Approach to Bacterial Dynamics during the Ripening of Spanish Farmhouse Cheese, Using Culture-Dependent and -Independent Methods

Martín‐Platero

Valdivia

Maqueda

et al. 2008

Appl Environ Microbiol

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We studied the dynamics of the microbial population during ripening of Cueva de la Magahá cheese using a combination of classical and molecular techniques. Samples taken during ripening of this Spanish goat's milk cheese in which Lactococcus lactis and Streptococcus thermophilus were used as starter cultures were analyzed. All bacterial isolates were clustered by using randomly amplified polymorphic DNA (RAPD) and identified by 16S rRNA gene sequencing, species-specific PCR, and multiplex PCR. Our results indicate that the majority of the 225 strains isolated and enumerated on solid media during the ripening period were nonstarter lactic acid bacteria, and Lactobacillus paracasei was the most abundant species. Other Lactobacillus species, such as Lactobacillus plantarum and Lactobacillus parabuchneri, were also detected at the beginning and end of ripening, respectively. Non-lactic-acid bacteria, mainly Kocuria and Staphylococcus strains, were also detected at the end of the ripening period. Microbial community dynamics determined by temporal temperature gradient gel electrophoresis provided a more precise estimate of the distribution of bacteria and enabled us to detect Lactobacillus curvatus and the starter bacteria S. thermophilus and L. lactis, which were not isolated. Surprisingly, the bacterium most frequently found using culture-dependent analysis, L. paracasei, was scarcely detected by this molecular approach. Finally, we studied the composition of the lactobacilli and their evolution by using length heterogeneity PCR.Lactic acid bacteria (LAB) are functionally related by their ability to produce lactic acid via either homo-or heterofermentative metabolism. The acidification and enzymatic processes accompanying the growth of LAB impart distinctive flavors and textures to a wide variety of fermented dairy products, meats, and vegetables. In addition, their preservative qualities make these bacteria useful in the control of undesirable microorganisms (16). For this reason LAB have important applications in the food, agricultural, and medical sectors and have been the subject of considerable research and commercial development over the past decade. Lactobacilli are well represented in fermented milk, yogurt, and cheese, as either natural or intentionally added microbiota (starter cultures). Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus curvatus, Lactobacillus acidophilus, and Lactobacillus pentosus are common members of the nonstarter LAB (NSLAB) communities in dairy products (11,32,35). In cheese NSLAB are represented mainly by L. casei and L. paracasei (5,11,20), but L. brevis, L. plantarum and L. curvatus are also important (4,5,43).

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Section: Discussionsupporting

confidence: 62%

Polyphasic Approach to Bacterial Dynamics during the Ripening of Spanish Farmhouse Cheese, Using Culture-Dependent and -Independent Methods

Martín‐Platero

Valdivia

Maqueda

et al. 2008

Appl Environ Microbiol

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“…PCR products were resolved on 2% agarose gel in TAE buffer at constant voltage of 80 V (8 V cm À1 ) for 2.5 h. and about the same cell densities continued throughout ripening. The relatively high NSLAB number, 6-8 log cfu g À1 , were observed in Norvegia cheese already after 30 days of ripening (Skeie, Lindberg, & Narvhus, 2001;Kjustad, 2002;Østlie, Eliassen, Florvaag, & Skeie, 2004).…”

Section: Discussionmentioning

confidence: 96%

Phenotypic and PCR-based characterization of the microflora in Präst cheese during ripening

Østlie

Eliassen

Florvaag

et al. 2005

International Dairy Journal

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“…Comparison of the gene sequences of bacterial species showed that the 16S rRNA gene is highly conserved within a species and among species of the same genus, so 16S rRNA gene can be used as the ''gold standard'' for the determination of the bacteria [5]. However, genotypic identification based on 16S rRNA sequences has limited discriminating power for closely related Lactobacillus species [6,7]. Recently, more protein encoding genes have been applied for classification of closely related species: the tuf gene (encoding elongation factor Tu) [8], the hsp60 gene (encoding 60-kDa heat shock protein) [9], the htrA gene (encoding stress-inducible trypsin-like serine protease) [10], the recA gene (encoding recombinase A) [11], and the rpoA gene (encoding RNA polymerase beta subunit) [12].…”

Section: Introductionmentioning

confidence: 99%

Phylogenetic study of Lactobacillus acidophilus group, L. casei group and L. plantarum group based on partial hsp60, pheS and tuf gene sequences

Sun

Liu

et al. 2012

Eur Food Res Technol

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The partial nucleotide sequences encoding elongation factor Tu (tuf gene), 60-kDa heat shock protein (hsp60 gene) and phenylalanyl-tRNA synthase (pheS gene) were determined to assess the suitability as phylogenetic markers for discriminating the closely related species in Lactobacillus acidophilus group, L. casei group and L. plantarum group. A total of 234 lactobacilli were chosen from traditional fermented dairy products that were not exactly assigned to species based on biochemical tests and 16S rRNA gene sequences. The sequencing of partial tuf, hsp60 and pheS gene of all strains was performed, and then, the phylogenetic trees were constructed by neighborjoining method. Phylogenetic tree revealed three genes provided better resolution of each Lactobacillus species than 16S rDNA, and all of strains were clearly identified as L. casei (63 strains), L. plantarum (58 strains) and L. helveticus (113 strains) by comparison of sequences with the type strains. From our results, the partial sequences of three genes had a higher discriminatory power than 16S rRNA gene sequences and were an alternative molecular tool for the taxonomical analysis of L. casei group, L. plantarum group and L. acidophilus group.

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Phenotypic and PCR-based characterization of the microflora in Norvegia cheese during ripening

Cited by 47 publications

References 35 publications

Polyphasic Approach to Bacterial Dynamics during the Ripening of Spanish Farmhouse Cheese, Using Culture-Dependent and -Independent Methods

Polyphasic Approach to Bacterial Dynamics during the Ripening of Spanish Farmhouse Cheese, Using Culture-Dependent and -Independent Methods

Phenotypic and PCR-based characterization of the microflora in Präst cheese during ripening

Phylogenetic study of Lactobacillus acidophilus group, L. casei group and L. plantarum group based on partial hsp60, pheS and tuf gene sequences

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