M.-N. Leclercq-Perlat

Controlled production of Camembert-type cheeses. Part I: Microbiological and physicochemical evolutions

A holistic approach of a mould cheese ripening is presented. The objective was to establish relationships between the different microbiological and biochemical changes during cheese ripening. Model cheeses were prepared from pasteurized milk inoculated with Kluyveromyces lactis, Geotrichum candidum, Penicillium camemberti and Brevibacterium linens under aseptic conditions. Two cheese-making trials with efficient control of environmental parameters were carried out and showed similar ripening characteristics. K. lactis grew rapidly between days 1 and 6 (generation time around 48 h). G. candidum grew exponentially between days 4 and 10 (generation time around 4.6 d). Brevi. linens also grew exponentially but after day 6 when Pen. camemberti mycelium began developing and the pH of the rind was close to 7. Its exponential growth presented 3 phases in relation to carbon and nitrogen substrate availability. Concentrations of Pen. camemberti mycelium were not followed by viable cell count but they were evaluated visually. The viable microorganism concentrations were well correlated with the carbon substrate concentrations in the core and in the rind. The lactose concentrations were negligible after 10 d ripening, and changes in lactate quantities were correlated with fungi flora. The pH of the inner part depended on NH3. Surface pH was significantly related to NH3 concentration and to fungi growth. The acid-soluble nitrogen (ASN) and non-protein nitrogen (NPN) indexes and NH3 concentrations of the rind were low until day 6, and then increased rapidly to follow the fungi concentrations until day 45. The ASN and NPN indexes and NH3 concentrations in the core were lower than in the rind and they showed the same evolution. G. candidum and Pen. camemberti populations have a major effect on proteolysis; nevertheless, K. lactis and Brevi. linens cell lysis also had an impact on proteolysis. Viable cell counts of K. lactis, G. candidum, Pen. camemberti and Brevi. linens were correlated with the environmental conditions, with proteolytic products and with carbon substrate assimilation. NH3 diffusion from surface to the cheese core during ripening was highly suspected. Interaction phenomena between microorganisms are discussed.

Growth of Debaryomyces hansenii on a bacterial surface-ripened soft cheese

Experimental cheeses were prepared in triplicate from pasteurized milk inoculated with Debaryomyces hansenii under aseptic conditions. Three cheesemaking replicates, with efficient control of environmental parameters (temperature, relative humidity, atmospheric composition) showed similar ripening characteristics. Deb. hansenii grew only on the cheese surface, where its oxygen demand was satisfied, especially during the first 24 h (mean generation time, 5·8 h). Salting in a sterile saturated brine solution reduced its growth and decreased viability. Growth was slower after 48 h because of the decrease in ripening temperature (mean generation time, 94 h). The total count of Deb. hansenii was maximum (≈3×10 7 yeasts/mm 2 ) after 6 d ripening and its viable cell concentration was ≈2×10 6 cfu/mm 2 . This difference was due to the ‘non-viability’ of part of the population. The viable Deb. hansenii concentration was highly correlated ( r 2 >0·95) with the lactate concentration in the inner part and with the surface and inner lactose concentrations, up to day 10 of ripening. This emphasized the importance of the diffusion of carbon substrate from the inner part to the surface of the cheese during ripening. The pH of the inner part depended significantly on the lactate and lactose concentrations. Surface pH was significantly related to inner lactate concentration, temperature and relative humidity. This also demonstrated the controlling role of carbon source diffusion.

Controlled production of camembert-type cheeses: part III role of the ripening microflora on free fatty acid concentrations.

Phenomena generating FFAs, important flavour precursors, are significant in cheese ripening. In Camembert-like cheeses, it was intended to establish the relationships between the dynamics of FFA concentrations changes and the succession of ripening microflora during ripening. Experimental Camembert-type cheeses were prepared in duplicate from pasteurised milk inoculated with Kluyveromyces lactis, Geotrichum candidum, Penicillium camemberti, and Brevibacterium aurantiacum under aseptic conditions. For each cheese and each cheesy medium, concentrations of FFAs with odd-numbered carbons, except for 9:0 and 13:0, did not change over time. For long-chain FFAs, concentrations varied with the given cheese part (rind or core). K. lactis produced only short or medium-chain FFAs during its growth and had a minor influence on caproic, caprylic, capric, and lauric acids in comparison with G. candidum, the most lipolytic of the strains used here. It generated all short or medium-chain FFAs (4:0-12:0) during its exponential and slowdown growth periods and only long-chain ones (14:0-18:0) during its stationary phase. Pen. camemberti produced more long-chain FFAs (14:0-18:0) during its sporulation. Brev. aurantiacum did not generate any FFAs. The evidence of links between specific FFAs and the growth of a given microorganism is shown.

Growth and colour development of some surface ripening bacteria with Debaryomyces hansenii on aseptic cheese curd.

The growth of five bacteria isolated from red-smear cheeses, Brevibacterium aurantiacum, Corynebacterium casei, Corynebacterium variabile, Microbacterium gubbeenense and Staphylococcus saprophyticus in mixed cultures with Debaryomyces hansenii on aseptic model cheese curd at 10 and 14 degrees C was investigated. At both temperatures, C. casei and Micro. gubbeenense had a longer lag phase than C. variabile, Brevi. aurantiacum and Staph. saprophyticus. In all cultures, lactose was utilised first and was consumed more rapidly at 14 degrees C than at 10 degrees C, i.e., 6 d at 14 degrees C and 10 d at 10 degrees C. This utilisation coincided with the exponential growth of Deb. hansenii on the cheese surface. Lactate was also used as a carbon source and was totally consumed after 21 d at 14 degrees C and approximately 90% was consumed after 21 d at 10 degrees C regardless of the ripening culture. Small differences (<0.5 pH unit) in the surface-pH during ripening were noticeable between ripening cultures. Differences in the colour development of the mixed cultures with the yeast control were only noticeable after 15 d for Brevi. aurantiacum and after 21 d for the other bacteria. Regardless of the organisms tested, colour development and colour intensity were also greater at 14 degrees C than at 10 degrees C. This study has provided useful information on the growth and contribution to colour development of these bacteria on cheese.

Controlled production of Camembert-type cheeses. Part II. Changes in the concentration of the more volatile compounds.

Flavour generation in cheese is a major aspect of ripening. In order to enhance aromatic qualities it is necessary to better understand the chemical and microbiological changes. Experimental Camembert-type cheeses were prepared in duplicate from pasteurized milk inoculated with Kluyveromyces lactis, Geotrichum candidum, Penicillium camemberti and Brevibacterium linens under aseptic conditions. Two replicates performed under controlled conditions of temperature (12 degrees C), relative humidity (95 +/- 2%), and atmosphere showed similar ripening characteristics. The evolutions of metabolite concentrations were studied during ripening. The volatile components were extracted by dynamic headspace extraction, separated and quantified by gas chromatography and identified by mass spectrometry. For each cheese the volatile concentrations varied with the part considered (rind or core). Except for ethyl acetate and 2-pentanone, the volatile quantities observed were higher than their perception thresholds. The flavour component production was best correlated with the starter strains. During the first 10 days the ester formations (ethyl, butyl and isoamyl acetates) were associated with the concentrations of K. lactis and G. candidum. The rind quantity of esters was lower than that observed in core probably due to (1) a diffusion from the core to the surface and (2) evaporation from the surface to the chamber atmosphere. G. candidum and Brev. linens association produced 3 methyl butanol and methyl 3-butanal from leucine, respectively. DMDS came from the methionine catabolism due to Brev. linens. Styrene production was attributed to Pen. camemberti. 2-Pentanone evolution was associated with Pen. camemberti spores and G. candidum. 2-Heptanone changes were not directly related to flora activities while 2-octanone production was essentially due to G. candidum. This study also demonstrates the determining role of volatile component diffusion.

First isolation of Brevibacterium sp. pigments in the rind of an industrial red-smear-ripened soft cheese

The smear-ripened soft cheeses are characterised by a surface orange-red-brown colour, which has a microbial origin. For a long time, this colouration was mainly imputed to Brevibacterium linens. However, the latest published works, based on molecular biology, have shown a minor role for this bacterium. This study shows the results obtained with an industrial cheese named Vieux-Pane, which is characterised by the presence of carotenoids from Brevibacterium linens group at its surface. This demonstrates that, under certain conditions, the Brevibacterium linens group (Brevibacterium linens and Brevibacterium aurantiacum sp. nov.) is able to produce pigments and to colour cheeses effectively.