Biodiversity of the Surface Microbial Consortia from Limburger, Reblochon, Livarot, Tilsit, and Gubbeen Cheeses

DOI: 10.1128/microbiolspec.CM-0010-2012 Publisher: ASM Press


Comprehensive collaborative studies from our laboratories reveal the extensive biodiversity of the microflora of the surfaces of smear-ripened cheeses. Two thousand five hundred ninety-seven strains of bacteria and 2,446 strains of yeasts from the surface of the smear-ripened cheeses Limburger, Reblochon, Livarot, Tilsit, and Gubbeen, isolated at three or four times during ripening, were identified; 55 species of bacteria and 30 species of yeast were found. The microfloras of the five cheeses showed many similarities but also many differences and interbatch variation. Very few of the commercial smear microorganisms, deliberately inoculated onto the cheese surface, were reisolated and then mainly from the initial stages of ripening, implying that smear cheese production units must have an adventitious "house" flora. Limburger cheese had the simplest microflora, containing two yeasts, Debaryomyces hansenii and Geotrichum candidum, and two bacteria, Arthrobacter arilaitensis and Brevibacterium aurantiacum. The microflora of Livarot was the most complicated, comprising 10 yeasts and 38 bacteria, including many gram-negative organisms. Reblochon also had a very diverse microflora containing 8 yeasts and 13 bacteria (excluding gram-negative organisms which were not identified), while Gubbeen had 7 yeasts and 18 bacteria and Tilsit had 5 yeasts and 9 bacteria. D. hansenii was by far the dominant yeast, followed in order by G. candidum, Candida catenulata, and Kluyveromyces lactis. B. aurantiacum was the dominant bacterium and was found in every batch of the 5 cheeses. The next most common bacteria, in order, were Staphylococcus saprophyticus, A. arilaitensis, Corynebacterium casei, Corynebacterium variabile, and Microbacterium gubbeenense. S. saprophyticus was mainly found in Gubbeen, and A. arilaitensis was found in all cheeses but not in every batch. C. casei was found in most batches of Reblochon, Livarot, Tilsit, and Gubbeen. C. variabile was found in all batches of Gubbeen and Reblochon but in only one batch of Tilsit and in no batch of Limburger or Livarot. Other bacteria were isolated in low numbers from each of the cheeses, suggesting that each of the 5 cheeses has a unique microflora. In Gubbeen cheese, several different strains of the dominant bacteria were present, as determined by pulsed-field gel electrophoresis, and many of the less common bacteria were present as single clones. The culture-independent method, denaturing gradient gel electrophoresis, resulted in identification of several bacteria which were not found by the culture-dependent (isolation and rep-PCR identification) method. It was thus a useful complementary technique to identify other bacteria in the cheeses. The gross composition, the rate of increase in pH, and the indices of proteolysis were different in most of the cheeses.

Download full-text


Available from: Nagamani Bora, May 06, 2014
  • [Show abstract] [Hide abstract]
    ABSTRACT: Cheese is a food which has been produced for centuries. While cheese was originally developed as a product which extended the shelf life of milk, over time distinct cheese varieties arose, being shaped by geographic, climate, cultural, and economic factors. Global demand for artisan cheeses is creating new economic opportunities. Consumers seeking distinctive products with regional flavor, or terroir, are becoming connoisseurs of hand-crafted cheeses with distinctive tastes and character. These demands have spurred new inquiry into microorganisms used as starter cultures and adjunct cultures, as well as the microbiological consortia of finished cheeses. Such demands have also created new concerns for food safety and international trade. New bacterial pathogens such as Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium DT104 have emerged in the food supply, causing a reevaluation of the efficacy of traditional cheesemaking procedures to control these pathogens. Similarly, pathogens such as Listeria monocytogenes pose problems to susceptible human populations, and cheese can be a vehicle of transmission for this deadly pathogen. With changes in sanitary requirements due to the globalization of the food industry, governments around the world are increasingly requiring assurances of cheese safety. While many governments recognize the safety of traditional artisan cheeses manufactured from raw milk, others are demanding pasteurization of all milk intended for cheesemaking to provide assurance of microbiological safety. In response, new technologies are being proposed to increase cheese safety, but these technologies fundamentally alter the traditional artisan practices and may not enhance microbiological safety. A reevaluation of the safety of traditional artisan practices, validation thereof, and communication of the scientific principles which promote safety will be necessary to enable the continued production of traditional artisan cheeses in global commerce. This also affords the opportunity to more fully explore the microbial diversity and microbial ecology of the great cheeses of the world.
    No preview · Article · Oct 2013
  • [Show abstract] [Hide abstract]
    ABSTRACT: Cheese ripening involves the activity of various bacteria, yeasts or molds, which contribute to the development of the typical color, flavor and texture of the final product. In situ measurements of gene expression are increasingly being used to improve our understanding of the microbial flora activity in cheeses. The objective of the present study was to investigate the physiology and metabolic activity of Geotrichum candidum during the ripening of Reblochon-type cheeses by quantifying mRNA transcripts at various ripening times. The expression of 80 genes involved in various functions could be quantified with a correct level of biological repeatability using a set of three stable reference genes. As ripening progresses, a decrease in expression was observed for genes involved in cell wall organization, translation, vesicular mediated transport, and in cytoskeleton constituents and ribosomal protein genes. There was also a decrease in the expression of mitochondrial F1F0 ATP synthase and plasma membrane H(+) ATPase genes. Some genes involved in the catabolism of lactate, acetate and ethanol were expressed to a greater extent at the beginning of ripening. During the second part of ripening, there was an increased expression of genes involved in the transport and catabolism of amino acids, which could be attributed to a change in the energy source. There was also an increase in the expression of genes involved in autophagy and of genes possibly involved in lifespan determination. Quantification of mRNA transcripts may also be used to produce bioindicators relevant for cheesemaking, for example when considering genes encoding enzymes involved in the catabolism of amino acids. Copyright © 2014 Elsevier B.V. All rights reserved.
    No preview · Article · Nov 2014 · International Journal of Food Microbiology