The microbiota of high-moisture mozzarella cheese produced with different acidification methods

The microbiota of high-moisture Mozzarella cheese made from cow's milk and produced with different acidification methods was evaluated at the end of refrigerated storage by pyrosequencing of the 16S rRNA gene. The cheeses were clearly separated on the basis of the acidification methods. Cheeses produced with the addition of starters were dominated by Streptococcus thermophilus, but a variety of lactic acid bacteria and spoilage microorganisms appeared at low levels (0.01-1%). Cheeses produced by direct addition of citric acid were dominated by a diverse microbiota, including both lactic acid bacteria and psychrotrophic γ-proteobacteria. For five brands the acidification system was not declared on the label: the microbiota was dominated by thermophilic lactic acid bacteria (S. thermophilus, Lactobacillus delbrueckii, Lactobacillus helveticus) but a variety of other subdominant lactic acid bacteria, psychrotrophs and Enterobacteriaceae were present, with a diversity comparable or higher to cheeses produced by direct acid addition. This led to the conclusion that undefined starters were used for acidification. Both ordination methods and network analysis were used for the representation of beta-diversity: matrix cluster analysis, principal coordinate analysis and OTU networks uncovered different aspects of the microbial community structure. For three cheese brands both biological replicates (cheeses from different lots) and technical replicates (replicate cheeses from the same lot) were analyzed. Repeatability was acceptable for OTUs appearing at frequencies >1%, but was low otherwise. A linear mixed model showed that the starter system was responsible for most differences related to dairies, while difference due to psychrotrophic contaminants was more related to lot-to-lot variability.