Abstract

Microorganisms are an essential component of cheeses and play important roles during both cheese manufacture and ripening. Both starter and secondary flora modify the physical and chemical properties of cheese, contributing and reacting to changes that occur during the manufacture and ripening of cheese. As the composition of microbial population changes under the influence of continuous shifts in environmental conditions and microorganisms interactions during manufacturing and ripening, the characteristics of a given cheese depend also on microflora dynamics. The microbiota present in cheese is complex and its growth and activity represent the most important, but the least controllable steps. In the past, research in this area was dependent on classical microbiological techniques. However, culture-dependent methods are time-consuming and approaches that include a culturing step can lead to inaccuracies due to species present in low numbers or simply uncultivable. Therefore, they cannot be used as a unique tool to monitor community dynamics. For these reasons approaches to cheese microbiology had to change dramatically. To address this, in recent years the focus on the use of culture-independent methods based on the direct analysis of DNA (or RNA) has rapidly increased. Application of such techniques to the study of cheese microbiology represents a rapid, sound, reliable, and effective way for the detection and identification of the microorganisms present in dairy products, leading to major advances in understanding this complex microbial ecosystem and its impact on cheese ripening and quality. In this article, an overview on the recent advances in the use of molecular methods for thorough analysis of microbial communities in cheeses is given. Furthermore, applications of culture-independent approaches to study the microbiology of two important raw-milk, long-ripened cheeses such as Grana Padano and Parmigiano Reggiano, are presented.

Highlights

  • Grana Padano (GP) and Parmigiano Reggiano (PR), the most widespread Italian cheese varieties, are raw-milk, long-ripened, hard cooked cheeses

  • Culture-independent methods applied to the study of the microbial ecology of GP and PR natural whey starters revealed that both are dominated by thermophilic lactic acid bacteria (LAB) belonging to Lb. helveticus, Lb. delbrueckii subsp. lactis, Lb. fermentum, and St. thermophilus species (Rossetti et al, 2008; Bottari et al, 2010)

  • LH-polymerase chain reaction (PCR) was used as consolidated reference culture-independent method in order to highlight the importance of using both the approaches to know the microbiota of raw-milk, long-ripened cheeses

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Summary

Introduction

Grana Padano (GP) and Parmigiano Reggiano (PR), the most widespread Italian cheese varieties, are raw-milk, long-ripened, hard cooked cheeses. LAB, from raw milk and natural whey starter (undefined thermophilic starter culture in whey), have a crucial role during both cheese-making and ripening. Rossetti et al (2008), for example, evaluated the species composition and the genotypic strain heterogeneity of dominant LAB isolated from natural whey starter cultures used to GP manufacture, by means of agar plate counting and reverse transcriptase, LH-PCR (RT-LH-PCR).

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