Possible implications of milk pasteurization on the manufacture and sensory quality of ripened cheese

Cheese made from raw milk represents an important proportion of the traditional cheeses, particularly in South European countries. Besides destruction of pathogenic bacteria, the most significant changes in milk relevant to cheesemaking, which are induced by pasteurization are:

• a partial elimination of the milk microorganisms which may grow in cheese during ripening,

• a partial or total activation or inhibition of the plasmin/plasminogen complex, cathepsin D, lipoprotein lipase and alkaline phosphatase. Enzymes from psychrotrophic bacteria, acid phosphatase and xanthine oxidase, which may be active during ripening, withstand pasteurization.,

• a slight (7%) denaturation of serum proteins and little or no modification of the cheesemaking properties (coagulation, acidification by lactic acid bacteria).

From experimental work carried out on several cheese varieties, comparing pasteurized or microfiltered milk and raw milk cheeses, it was found that facultatively heterofermentative lactobacilli, Micrococcaceae, enterococci, and propionibacteria in Swiss-type cheese, are found at higher levels in raw milk cheese. The main biochemical modification of cheese during ripening concerns the nature and extent of proteolysis. Although there is no clear trend in the breakdown of _s1- and β-caseins, milk pasteurization leads to a significant decrease of the amount of small peptides and free amino acids and to different HPLC profiles. Experiments carried out with sensory analysis show that, in all cases, pasteurized or microfiltered milk cheeses have received lower flavour intensity scores than raw milk cheeses. From this review, it is concluded that the indigenous milk microflora, with its diversity of species and strains, appears to be mainly responsible of the specific sensory properties of raw milk cheeses.     

Fate of Listeria innocua during production and ripening of smeared hard cheese made from raw milk

The fate of 2 different Listeria innocua strains was analyzed during the production and ripening of smeared raw milk Greyerzer cheese (Gruyère). These strains were used as surrogates for the pathogenic Listeria monocytogenes, as they are physiologically very similar. Bacterial cells were added to the cheese milk at levels of 10⁵ cfu/mL. During the first 24 h of cheese making, the number of the test strains decreased to a level of below 10² cfu/g. Obviously, the cooking temperature of 56°C and the subsequent slight temperature decrease to 50°C within 70 min contributed to a distinct reduction of Listeria counts. The counts in the cheese cores did not exceed 10³ cfu/g within 12 wk of cheese ripening and Listeria was not detectable after 24 wk. In contrast to the cores of the cheeses of the 4 batches in this study, their rinds always contained a high listerial load of approximately 10⁶ to 10⁸ cfu/g throughout the entire ripening period. The smeared surface showed an increase of pH to alkaline values, corresponding to smear microbiota development. Coryneforms and Staphylococcus counts were stable at >10⁷ cfu/cm² over 175 d, whereas yeast counts decreased to about 10⁵ cfu/cm² at the end of ripening. The study shows that the smear culture had no noticeable anti-listerial potential. When removing the rind or portioning such smeared cheese loaves with a cutting device, a postprocess contamination of the core might occur, thus presenting a major hygienic risk.     

Listeria monocytogenes survival during production and storage of water buffalo Mozzarella cheese

The ability of Listeria monocytogenes to survive during the manufacture of water buffalo Mozzarella and to grow during its shelf life was evaluated. A wild‐type and a reference strain were used to contaminate raw milk. The viable count of the reference strain ATCC 9525 dropped after the stretching process, and in the cheese, it fell to below 100 cfu/g. When the wild‐type strain was used, however, stretching did not appear to have any effect on the pathogen. The artificially contaminated cheeses were stored, for eleven days, at 4, 20 and 30 °C. Pathogen populations increased at 20 (≈2.60 log cfu/g) and 30 °C (≈1.95 log cfu/g).     

Behavior of Shiga-toxin-producing Escherichia coli in ewe milk stored at different temperatures and during the manufacture and ripening of a raw milk sheep cheese (Zamorano style)

This study was conducted to assess the survival of 2 wild Shiga toxin-producing Escherichia coli strains (one serotype O157:H7 and one non-O157:H7) in ewe milk stored at different conditions and to examine the fate of the O157 strain during the manufacture and ripening of a Spanish sheep hard variety of raw milk cheese (Zamorano). The strains were selected among a population of 50 isolates, which we obtained from ewe milk, because of their high resistance to 0.3% lactic acid. Both strains were inoculated (approximately 2 log₁₀ cfu/mL) in raw and heat-treated (low-temperature holding, LTH; 63°C/30 min) ewe milk and stored for 5 d at 6, 8, and 10°C and also according to a simulation approach for assessing the effects of failures in the cold chain. The minimum growth temperature for the O157:H7 strain in LTH and raw ewe milk was 8°C. For the non-O157:H7 strain, the lowest temperature showing bacterial growth in LTH ewe milk was 6°C, but it did not grow at any of the tested conditions in raw milk. It appears that the O157 strain was more susceptible to cold stress but was likely a better competitor than the non-O157 strain against the milk autochthonous microbiota. For manufacture of Zamorano cheese, raw milk was inoculated with approximately 3 log₁₀ cfu/mL, and after 2 mo of ripening at 10 to 12°C, the cheeses showed the expected general characteristics for this variety. The O157:H7 strain increased 0.9 log₁₀ cfu/g after whey drainage and during ripening and storage decreased by 2.9 log₁₀ cfu/g. Nevertheless, its detectable level (estimated at 6.2 cfu/g) after 2 mo of ripening suggests that Zamorano cheese manufactured from raw ewe milk contaminated with E. coli O157:H7 could represent a public health concern.     

Effect of wild strains of Lactococcus lactis on the volatile profile and the sensory characteristics of ewes' raw milk cheese

The production of volatile compounds by wild strains of Lactococcus lactis used as starter cultures and their effect on the sensory characteristics of ewes' raw milk cheese were investigated. Sixteen vats of cheese were manufactured and ripened for 120 d in two experiments, each of them duplicated. In the first experiment, milk was inoculated with different ratios of four wild Lactococcus lactis strains, two producing and two not producing branched-chain volatile compounds, and in the second experiment with different ratios of a commercial starter culture and the two strains producing branched-chain volatile compounds. Cheese pH, proteolysis, and aminopeptidase activity increased when the strains producing branched-chain volatile compounds were inoculated at a higher rate. Fifty volatile compounds were identified in cheeses using a purge and trap system coupled to a gas chromatography-mass spectrometry apparatus. The relative abundances of 30 volatile compounds (8 alcohols, 5 aldehydes, 3 ketones, 12 esters, 1 sulfur compound, and 1 benzenic compound) were influenced by starter culture composition. 2-Methylpropanol, 3-methylbutanol, isobutyl acetate, isoamyl acetate, ethyl butyrate, isobutyl butyrate, and isoamyl butyrate were always more abundant in the cheeses made with a higher level of L. lactis strains producing branched-chain volatile compounds. Flavor intensity was enhanced by a high level of L. lactis strains producing branched-chain volatile compounds in the first experiment, in which four wild L. lactis strains were used as starter culture, but not in the second experiment, in which a combination of two wild L. lactis strains and the commercial starter culture were used. Flavor quality, as judged by trained panelists, was impaired in both experiments by a high level of L. lactis strains producing branched-chain volatile compounds.     

Microbial community dynamics of a blue-veined raw milk cheese from the United Kingdom

A commercial blue-veined cheese made from unpasteurized milk was examined by conventional culturing and PCR denaturing gradient gel electrophoresis analysis of the bacterial community 16S rRNA genes using 3 primer sets, V3, V4V5, and V6V8. Genomic DNA for amplification was extracted directly from raw milk, starter culture, cheese at different stages of production, fully ripened cheese, and from the cultured cells grown on various media. The outer rind was sampled separately from the inner white core and blue veins. A diverse microbiota containing Lactococcus lactis ssp. lactis, Lactobacillus plantarum, Lactobacillus curvatus, Staphylococcus gallinarum, Staphylococcus devriesei, Microbacterium sp., Sphingobacterium sp., Mycetocola sp., Brevundimonas sp., Enterococcus faecalis, Proteus sp., and Kocuria sp. was detected in the raw milk using culturing methods, but only Lactococcus lactis ssp. lactis, Lactobacillus plantarum, and Enterococcus faecalis survived to the final cheese and were detected both in the core and the rind. Using PCR denaturing gradient gel electrophoresis analysis of the cheese process samples, Staphylococcus equorum and Enterococcus durans were found in the rind of prepiercing samples but not in the core and veins; after piercing, these species were found in all parts of the cheese but survived only in the rind when the cheese was fully ripened. Brevibacterium sp., Halomonas sp., Acinetobacter sp., Alkalibacterium sp., and Corynebacterium casei were identified only by PCR denaturing gradient gel electrophoresis and not cultured from the samples. Brevibacterium sp. was initially identified in the cheese postpiercing (core and veins), Halomonas sp. was found in the matured cheese (rind), and Acinetobacter sp., Alkalibacterium sp., and Corynebacterium casei were also found in the prepiercing samples (rind) and then found through the subsequent process stages. The work suggests that in this raw milk cheese, a limited community from the milk survive to the final cheese, with salt addition and handling contributing to the final cheese consortium.     

不同贮存条件对原料乳质量的影响

研究了原料乳质量和贮存温度及贮存时间之间的关系.通过菌落总数的检测和酸度的测定,探讨了不同贮存温度随时间对原料乳中菌落总数和酸度的影响,旨在为优质原料乳的贮运提供参考.结果表明,在4℃贮存条件下,原料乳存放36 h,对质量没有较大的影响;在原料乳温度达到10～15℃的时候,存放时间不宜超过12 h.     

Effect of high‐pressure treatments on microbial loads and physicochemical characteristics during refrigerated storage of raw milk Serra da Estrela cheese samples

This work studied the effect of high pressure processing (HPP) at 400, 500 and 600 MPa during 10, 5 and 3 min, respectively, on samples ewe cheese manufactured from raw milk, during storage (100 days) at 5 °C. Total aerobic mesophilic and lactic acid bacteria were slightly affected, decreasing by about 1.0 and 0.82 log CFU g^?1, respectively, immediately after HPP treatment at 600 MPa for 3 min, while Enterobacteriaceae, yeasts and moulds, and Listeria innocua were reduced to below the quantification limits. Lactic acid bacteria decreased further during storage, showing increasing inactivation as the pressure level increased. Physicochemical parameters (water activity, moisture content, pH and titratable acidity) were generally not affected by HPP, while lipid oxidation increased throughout storage, with HPP samples showing lower values (50–66%) at 100 days of storage. The results indicated that HPP has potential to improve cheese microbial safety and shelf‐life, with a lower lipid oxidation level than nonpressurised cheese.     

生产贮藏条件对原料奶中微生物的影响

以手工挤奶条件下不同来源的原料奶为研究对象,在冷藏(4℃)、室温(19℃)条件下,分别贮藏0,3,6,9 h后检测原料奶中菌落总数(TBC)、芽孢总数(TSC)、耐热芽孢数、嗜冷菌总数。结果表明,随着贮藏温度增加、时间延长,原料奶中微生物增幅加快(P<0.05),冷链条件下(4℃)贮运原料奶对控制微生物生长繁殖具有重要作用。手工挤奶条件下室温(19℃)贮奶3h、冷藏(4℃)贮奶6h后原料奶中菌落总数超过5×105 mL-1的国家标准,原料奶的芽孢总数、耐热芽孢数和嗜冷菌数均超出液态奶的生产要求,特别不利于长效奶(保质期30 d以上)的生产。改善牛奶生产环境对提高原料奶品质具有重要的作用。     

The effect of refrigerated storage of raw milk on the physicochemical and microbiological quality of Tunisian semihard Gouda‐type cheese during ripening

A Tunisian semihard Gouda‐type cheese made from milk kept at 4 °C for 24, 48, 72 and 96 h was monitored during 45 days of ripening. The effect of milk refrigeration on the evolution of physicochemical parameters in relation to the quantitative variation of the microbial population during ripening of Gouda‐type cheese was investigated. Microbiological and physicochemical analyses were performed on raw milk and cheese samples after curding, 2, 9, 16, 23, 30, 37 and 45 days of ripening time. The raw milk kept under refrigeration at 4 °C for 96 h showed the highest microbial count and proteolysis level. The duration of storage significantly reduced the cheese yield as a result of important solubilisation casein in proteoses‐peptones. Results of different nitrogenous fractions by Kjeldahl method showed enzymatic hydrolysis products of casein whose intensity depended on the maturing stage as well as the refrigeration time. Besides the evident action of the plasmin, original milk protease, on the hydrolysis of casein in soluble fractions, the proteolysis of cheese caseins is also initiated by proteolytic action of the chymosin and extracellular heat‐resistant proteases notably produced by the same psychrotrophic microflora. Lactic acid bacteria starters that constitute the dominant microflora of this type of cheese are also considered as aroma precursors.     

水牛乳和荷斯坦牛乳切达干酪成熟期间质量特性对比

为了了解水牛乳和荷斯坦牛乳切达干酪成熟期间的质量特性,采用了理化测定、质构测定和感官评定相结合的方法对其进行研究.结果表明:在成熟期90 d内,不同成熟温度下,两种干酪的蛋白质质量分数减少约为1％～4％,脂肪质量分数减少约为4％～10％,且成熟温度越高,质量分数下降越大,水牛乳干酪中蛋白和脂肪质量分数均高于荷斯坦牛乳干酪且两种成分的下降量也是水牛乳干酪更高;pH值呈现先下降后上升趋势.在90 d成熟期间,两种干酪的硬度先上升后下降,凝聚性上升,弹性下降.经感官评价,两种干酪在10℃成熟60天时风味最佳,水牛乳切达干酪略受偏爱.可用于水牛乳切达干酪成熟期内质量控制.     

The occurrence of Staphylococcus aureus on a farm with small-scale production of raw milk cheese

In recent years, the small-scale production of raw milk products has increased in Norway, and there is some concern that such foods may pose a risk of staphylococcal food poisoning to consumers. The aim of the study was to evaluate potential sources of contamination of raw milk cheese with Staphylococcus aureus on a bovine dairy farm with small-scale production. Samples for bacteriological analyses (n = 144) were collected from the animals, the environment, processing equipments, from humans, and from cheeses at various stages of production. Staphylococcus aureus was isolated from 10 of 11 cows, the farmer, equipment, the environment, and the cheese. Seventy-five Staph. aureus isolates were genotyped by pulsed-field gel electrophoresis, tested for enterotoxin (SE) production by reversed passive latex agglutination, for SE genes by multiplex polymerase chain reaction, and for penicillin resistance by the cloverleaf method. Five different pulsotypes were identified and SE gene fragments were identified in 11 isolates, but no isolates produced SE or were penicillin resistant. Staphylococcus aureus was found throughout the farm, and appeared to be spread with the milk to the environment, equipment, and to products. One pulsotype dominated and was identified from most sample sites on the farm. Raw milk products are vulnerable to contamination with Staph. aureus. Strategies to reduce the occurrence of Staph. aureus in bulk milk are of particular importance on farms where milk is used for raw milk products.     

Biochemical changes during the ripening of homemade 'San Simón da Costa' raw milk cheese

'San Simón da Costa' cheese is a traditional smoked variety produced in the northwest of Spain from cow's milk. Biochemical changes were determined during its ripening. Its high calcium and phosphorus content and its low NaCl and sodium content stand out. This cheese undergoes moderate proteolysis. The most abundant free amino acid at the end of the ripening was glutamic acid, followed by tryptophan, leucine, arginine and phenylalanine. The lipolysis throughout ripening is slight; the most abundant free fatty acid being oleic, followed by palmitic and butyric acid.     

Traditional Colonial-type cheese from the south of Brazil: A case to support the new Brazilian laws for artisanal cheese production from raw milk

Artisanal Colonial-type cheese is made from raw milk and is the main cheese produced by rural families of the southern region of Brazil. The aim of this study was to investigate, identify problems, and propose solutions for the current situation of small family farms producing and informally selling artisanal Colonial-type cheese located in the western part of Santa Catarina State in Southern Brazil. A semistructured questionnaire was employed in 12 rural properties to analyze the mode of production. Physical-chemical and microbiological analyses of water, raw milk, and cheese were performed, and it was found that 92, 50, and 100% of the samples, respectively, were outside of the current Brazilian regulatory parameters. None of the cheesemakers involved in this study met the requirements, as established by law, for artisanal cheese production from raw milk. This study concluded that technical support and changes in public policy are needed to ensure the preservation of this artisanal cheese, considering the historical importance and cultural traditions of these local communities and the socioeconomic importance of cheesemaking to family farming. Furthermore, more research on the safety of the cheese produced from raw milk is needed as well as the development of specific microbiological standards for artisanal Brazilian cheeses. Public policies aimed at guaranteeing food safety that formalize the commercialization of these cheeses will increase food security in those communities that currently produce artisanal cheese informally.     

The fate of Listeria monocytogenes during the manufacture of Camembert-type cheese: A comparison between raw milk and milk treated with high hydrostatic pressure

Camembert-type cheese was produced from: raw bovine milk; raw milk inoculated with 2 or 4 log CFU/ml Listeria monocytogenes; raw milk inoculated with L. monocytogenes and subsequently pressure-treated at 500 MPa for 10 min at 20 °C; or uninoculated raw milk pressure-treated under these conditions. Cheeses produced from both pressure-treated milk and untreated milk had the typical composition, appearance and aroma of Camembert. Curd and cheese made from inoculated, untreated milk contained large numbers of L. monocytogenes throughout production. An initial inoculum of 1.95 log CFU/ml in milk increased to 4.52 log CFU/g in the curd and remained at a high level during ripening, with 3.85 log CFU/g in the final cheese. Pressure treatment inactivated L. monocytogenes in the raw milk at both inoculum levels and the pathogen was not detected in any of the final cheeses produced from pressure-treated milk. Therefore high pressure may be useful to inactivate L. monocytogenes in raw milk that is to be used for the production of soft, mould-ripened cheese.

Industrial relevance

This paper demonstrates the potential of high pressure (HP) for treatment of raw milk to be used in the manufacture of soft cheeses. HP treatment significantly reduced the level of Listeria monocytogenes in the raw milk and so allowed the production of safer non-thermally processed camembert-like soft cheese.     

Physicochemical, microbiological and sensory changes during ripening and storage of Xinotyri, a traditional Greek cheese from raw goat's milk

Physicochemical, microbiological and sensory characteristics, evolution of lipolysis monitored by measuring acid degree value (ADV) and the main mineral elements were studied during ripening and storage of artisanal Xinotyri cheese from raw goat's milk. Cheeses were characterized by a high content in total solids (TS; 83%) and fat (59% of TS), and very low pH (4.0), water activity (0.87) and moisture (17%). Protein and salt contents at the end of storage were 31% and 2.8%, respectively. Lipolysis increased, while variations in mineral (Ca, P, Mg and Zn) content were found during ripening and storage. Cheeses were free of Salmonella and Listeria in 25 g, while enterobacteria, pseudomonads, and coagulase-positive staphylococci were < 100 cfu/g. Mesophilic lactic acid bacteria increased above 8 log cfu/g by day 6, but declined by 2–3 logs in the ripened cheese (45 days) and by 3–4 logs during cheese storage at 4°C for up to 180 days.     

The effect of raw milk microbial flora on the sensory characteristics of Salers-type cheeses

The sensory characteristics of Salers Protected Denomination of Origin raw-milk cheeses are linked to the biochemical composition of the raw material (milk) and to the resultant microbial community. To evaluate the influence of the microbial community on sensory characteristics, Salers-type cheeses were manufactured with the same pasteurized milk, reinoculated with 3 different microbial communities from 3 different filtrates from microfiltered milks. Each cheese was subjected to microbial counts (on selective media), biochemical tests, and volatile and sensory component analyses at different times of ripening. Adding different microbial communities to specimens of the same (biochemically identical) pasteurized milk lead to different sensory characteristics of the cheeses. Cheeses with fresh cream, hazelnut, and caramel attributes were opposed to those with fermented cream, chemical, and garlic flavors. The aromatic compounds identified (esters, acids, alcohols, and aldehydes) in these cheeses were quite similar. Nevertheless, one milk was distinguished by a higher content of acetoin, and lower 2-butanone and 3-methylpentanone concentrations. Over the production period of 1 mo, the different cheeses were characterized by the same balance of the microbial population assessed by microbial counts on different media. This was associated with the stability of some sensory attributes describing these cheeses. Nevertheless, there was no linear correlation between microbial flora data and sensory characteristics as measured in this study.     

不同贮存条件对生牛奶品质影响的研究

从对生牛奶品质影响较大的贮存温度和时间着手，探讨了不同贮存条件对生牛奶中细菌总数以及组成的影响，旨在为优质生牛奶的贮运提供参考。     

Proteolytic and lipolytic changes during the ripening of León raw cow's milk cheese, a Spanish traditional variety

Proteolytic and lipolytic changes were studied throughout ripening of five batches of León cow's milk cheese, a traditional variety made in the north of Spain. Total soluble nitrogen, non-protein nitrogen, oligopeptides nitrogen, amino nitrogen and ammonia nitrogen fractions increased slightly during the ripening process. The final values of these nitrogen fractions indicate that this cheese undergoes a very slight proteolysis as much in extent as in depth. This weak protein degradation is corroborated when the caseins and their degradation products were quantified by electrophoresis. β-Casein stayed practically intact throughout the ripening process and only 10% of α_s-casein became degraded. The content of total free amino acids increased progressively but in a slightly increased way during ripening, reaching final average values of 592 mg (100 g)⁻¹ of total solids. The most abundant free amino acid at the end of ripening was lysine, followed by leucine, glutamic acid, tryptophan, valine and phenylalanine. The acidity index of the fat values increased during ripening by a factor of 4.39. The final values of this parameter are in the range of those observed in other cow's milk cheeses ripened by bacteria. The content in total free fatty acids underwent an increase throughout ripening reaching final average values of 6669 ppm. The most abundant free fatty acid at the end of ripening was oleic acid followed by butyric and palmitic acids. The high content of short-chain fatty acids is outstanding, specially that of butyric acid.     

Effect of frozen storage on the proteolytic and rheological properties of soft caprine milk cheese

Freezing and long-term frozen storage had minimal impact on the rheology and proteolysis of soft cheese made from caprine milk. Plain soft cheeses were obtained from a grade A goat dairy in Georgia and received 4 storage treatments: fresh refrigerated control (C), aged at 4°C for 28 d; frozen control (FC), stored at −20°C for 2 d before being thawed and aged in the same way as C cheese; and 3-mo frozen (3MF), or 6-mo frozen (6MF), stored at −20°C for 3 or 6 mo before being thawed and aged. Soft cheeses had fragile textures that showed minimal change after freezing or over 28 d of aging at 4°C. The only exceptions were the FC cheeses, which, after frozen storage and aging for 1 d at 4°C, were significantly softer than the other cheeses, and less chewy than the other frozen cheeses. Moreover, after 28 d of aging at 4°C, the FC cheeses tended to have the lowest viscoelastic values. Slight variation was noted in protein distribution among the storage treatment, although no significant proteolysis occurred during refrigerated aging. The creation and removal of ice crystals in the cheese matrix and the limited proteolysis of the caseins showed only slight impact on cheese texture, suggesting that frozen storage of soft cheeses may be possible for year-round supply with minimal loss of textural quality.