Growth and metabolic activity of a cheese starter in CO2-acidified and non-acidified refrigerated milk

 The growth and activity of two Lactococcus strains and one Leuconostoc strain in CO₂-acidified and non-acidified refrigerated milk were evaluated separately and as a mixed culture to determine their suitability for use as a starter in the manufacture of Afuega’l Pitu, an acid-coagulated Spanish cheese, from refrigerated CO₂-acidified milk. The growth of the strains studied and their production of organic acids were similar in CO₂-acidified and non-acidified refrigerated or fresh milk, indicating that CO₂ treatment does not affect the metabolic activity of the strains. However, refrigeration enhanced the production of acetaldehyde, ethanol and diacetyl in CO₂-acidified and non-acidified milk. The level of diacetyl was also greater in refrigerated CO₂-acidified milk than in refrigerated non-acidified milk. It was concluded that refrigerated milk acidified with CO₂ can be satisfactorily used in the manufacture of Afuega’l Pitu cheese, and that this technique can be also used in the production of other acid-coagulated cheeses. Received: 1 July 1997 / Revised version: 1 September 1997     

Growth and metabolic activity of a cheese starter in CO2-acidified and non-acidified refrigerated milk

 The growth and activity of two Lactococcus strains and one Leuconostoc strain in CO₂-acidified and non-acidified refrigerated milk were evaluated separately and as a mixed culture to determine their suitability for use as a starter in the manufacture of Afuega’l Pitu, an acid-coagulated Spanish cheese, from refrigerated CO₂-acidified milk. The growth of the strains studied and their production of organic acids were similar in CO₂-acidified and non-acidified refrigerated or fresh milk, indicating that CO₂ treatment does not affect the metabolic activity of the strains. However, refrigeration enhanced the production of acetaldehyde, ethanol and diacetyl in CO₂-acidified and non-acidified milk. The level of diacetyl was also greater in refrigerated CO₂-acidified milk than in refrigerated non-acidified milk. It was concluded that refrigerated milk acidified with CO₂ can be satisfactorily used in the manufacture of Afuega’l Pitu cheese, and that this technique can be also used in the production of other acid-coagulated cheeses. Received: 1 July 1997 / Revised version: 1 September 1997     

Quality of plain yoghurt made from refrigerated and CO2-treated milk

The suitability of milk preserved by refrigeration and CO₂ addition for the manufacture of plain yoghurt was evaluated using two commercial strains of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus. Yoghurts manufactured, after milk pasteurisation, from refrigerated CO₂-treated samples (pH 6.15) were compared with two different controls made from pasteurised milk, either fresh or refrigerated. The multiplication and acidification capacity of the starter as well as the evolution of organic acids were not affected by the previous refrigeration and CO₂-treatment of raw milk nor by the residual CO₂ present in pasteurised milk. However, refrigeration enhanced the production of ethanol and diacetyl. No differences on sensory properties were detected through the cold storage between yoghurts made from CO₂-treated milk and those made from refrigerated control milk. It was concluded that refrigerated milk acidified with CO₂ could be satisfactorily used in the manufacture of yoghurt.     

Texture and flavour development in Ras cheese made from raw and pasteurised milk

Texture, proteolysis and flavour development in Ras cheeses made from raw or pasteurised milk with two different thermophilic lactic cultures were monitored during ripening. Results showed that at day 1 of manufacture, the moisture content and pH were lower in raw milk cheese than in pasteurised milk cheeses. Levels of water-soluble nitrogen, casein breakdown, free amino groups and free fatty acids were higher in cheese made from raw milk than in that made from pasteurised milk. Textural characteristics, such as hardness, cohesiveness and chewines, increased in all treatments during the first 60 days of ripening due to the reduction in the moisture level during the second stage of salting (dry salting during the first 60 days of ripening). Cheese made from raw milk received the highest texture and flavour scores by panellists.     

Manufacture of cheese made from CO2-treated milk

Cheeses were manufactured from pasteurised milk (control), pasteurised milk acidified to pH 6.0 with CO₂, and milk acidified to pH 6.0 with CO₂ prior to pasteurisation. Production of cheese from CO₂-treated milk at pH 6.0 reduced the amount of rennet necessary for coagulation by about 75%. Although acidification reduces the amount of lactic acid produced by starter during incubation of milk, no significant differences in lactic acid content were detected between cheeses manufactured from non-acidified or CO₂-acidified milks. Cheeses produced from CO₂-treated milk showed less proteolysis than control cheeses, but no significant differences in sensory characteristics between cheeses were detected.     

Influence of pasteurised milk, raw milk and different ripening cultures on biogenic amine concentrations in semi-soft cheeses during ripening

In semi-soft cheeses, produced with pasteurised milk, raw milk and different starter cultures, the concentrations of cadaverine, histamine, phenylethylamine, putrescine and tyramine were investigated. The cultures (pasteurised milk cultures, raw milk cultures and starter cultures) strongly influenced the biogenic amine concentrations in the cheeses ripened for 5 months. Two cheeses made with identical pasteurised milk, but different ripening cultures, differed greatly in their total biogenic amine concentrations (51 vs 371?mg/kg). In general, the biogenic amine concentrations increased markedly between month 2 and month 3 of cheese ripening. The high content of enterococci and Enterobacteriaceae yielded the biogenic amine concentrations. In contrast, Lactobacilli did not seem to be important. However, unspecified bacteria have to be considered, since cheeses with comparable microbiological profiles differed enormously in their biogenic amine concentrations. Semi-soft cheeses produced from pasteurised milk showed remarkably lower total biogenic amine concentrations compared to semi-soft cheeses produced from raw milk (51–1096?mg/kg vs 1011–3133?mg/kg, depending also on the ripening cultures). The highest total biogenic amine concentration (4817?mg/kg) was detected in a cheese produced from raw milk that had been stored for 36?h. In this cheese, the concentrations of cadaverine, phenylethylamine, putrescine and tyramine were higher than in all other cheeses. The highest histamine concentration was found to be in another raw milk cheese (573?mg/kg).     

Determination of bovine lactoferrin concentrations in cheese with specific monoclonal antibodies

In order to determine bovine lactoferrin concentration in cheese, bovine lactoferrin-specific monoclonal antibodies have been raised and an ELISA has been developed to determine lactoferrin concentrations in milk, whey and experimental soft, semi-hard and Swiss-type cheeses made with raw or pasteurised milk. The lactoferrin concentration in cheese was shown to depend on the cheese-making process, with higher values in Swiss-type and semi-hard cheeses than in soft cheeses. Furthermore, Western-blotting analysis of lactoferrin in cheese showed that this protein stayed intact throughout ripening in raw milk cheese, whereas it was partially hydrolysed in cheeses made with pasteurised milk. Based on these observations, we propose that cheese may constitute a natural dairy source of lactoferrin beneficial to health.     

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.     

Terpene profiles in Cantal and Saint‐Nectaire‐type cheese made from raw or pasteurised milk

Terpene profiles in cheese can be considered a ‘terroir’ fingerprint as the information contained in it should enable the pastures on which the animals were fed to be recognised. Yet a certain elasticity of the signature must be taken into account when determining authentication strategies, since products acknowledged as containing a common signature may have undergone certain procedures, such as cheese making and milk pasteurisation, that could have potentially altered their terpene profiles. In this study, Cantal and Saint‐Nectaire‐type cheeses were made from both raw and pasteurised milk from the same herd of dairy cows that had been grazed on natural grassland. Cheeses from raw and pasteurised milk were made from the same milking on the same days. Cantal and Saint‐Nectaire‐type cheeses were made on 4 different days, alternatively over four weeks. The terpenes in the cheese fat were analysed by dynamic headspace/gas chromatography/mass spectrometry. A great diversity of monoterpenes, sesquiterpenes and oxygen‐containing derivatives were identified. The major terpenes identified in most cheeses were β‐caryophyllene, α‐ and β‐pinene and limonene. Milk pasteurisation did not induce changes in the terpene profile of the cheese. Significant differences (p < 0.001) were observed between Cantal and Saint‐Nectaire cheeses: α‐pinene, β‐myrcene and β‐phellandrene were, respectively, three, five and five times more abundant in Cantal cheese, while tricyclene, α‐phellandrene and geraniol were found exclusively in Cantal cheese. In contrast, unidentified sesquiterpenes with retention indices (KI) = 1342 and 1511, α‐cubebene, longifolene and γ‐elemene were more abundant or exclusively found in Saint‐Nectaire cheese. A significant relationship with the date of milking (p < 0.01) was observed for α‐pinene and tricyclene in Cantal, for β‐myrcene, δ‐3‐carene, p‐cymene and α‐terpinene in Saint‐Nectaire cheese. Copyright © 2005 Society of Chemical Industry     

The fate of added nitrate used in the manufacture of Emmental cheese

The aim of this study was to observe the dynamics of nitrates and nitrites during the six stages of manufacture of Emmental cheeses. Samples were taken of untreated milk, of pasteurized milk, of milk with nitrates added, of pressed cheese curd, of whey, of maturing cheese and of the final product. The samples were drawn from a commercial operation in a cheese factory in the eastern part of Slovakia. The mean NaNO₂ content in untreated and in pasteurised milk was 0.2 and 0.1 mg kg^-1, respectively and the mean NaNO₃ content was 0.9 and 0.9 mg kg^-1 respectively. Nitrates were added to the milk to prevent 'blowing' of hard cheese by micro organisms. In milk with nitrate added the mean content was 81.2 mg kg^-1 NaNO₃; the maximum value being 90.0 mg kg¹ NaNO₃. After pressing, the mean value of nitrate was found to be 20.6 mg kg^-1 NaNO₃. A considerable quantity of nitrates passed into the whey, where the mean nitrate content was 67.0 mg kg^-1 NaNO₃. The final product had a markedly decreased content of nitrates (3.3 mg kg^-1 NaNO₃) and nitrites (0.2 mg kg^-1 NaNO₂) when compared with the values in cheese during maturation (11.3 mg kg^-1 NaNO₃; 0.4 mg kg^-1 NaNO₂).     

Comparison of biogenic amine profile in cheeses manufactured from fresh and stored (4 degrees C, 48 hours) raw goat's milk

In this study, the evolution of microbial counts, biogenic amine contents, and related parameters (pH, moisture, and proteolysis) in goat cheese made from fresh raw milk or raw milk stored for 48 h at 4 degrees C was examined. In both cases the milk was nonpasteurized. This study was designed to evaluate the effect of milk quality on the profile of biogenic amines in relation to the evolution of the microbial population during cheese making. Cheese made from raw milk stored for 48 h at 4 degrees C showed the highest microbial counts and biogenic amine levels. The storage of milk under refrigeration caused significant increases in the levels of some microbial and biogenic amines during ripening, but not initially. Tyramine was the main biogenic amine in the two cheeses tested, followed by cadaverine. However, the main differences in amine contents between batches were found for putrescine, histamine, and beta-phenylethylamine, whose levels were more than twofold higher in samples from raw milk refrigerated for 48 h than in samples from fresh milk.     

Effects of standardization of whole milk with dry milk protein concentrate on the yield and ripening of reduced-fat cheddar cheese

Commercial milk protein concentrate (MPC) was used to standardize whole milk for reduced-fat Cheddar cheesemaking. Four replicate cheesemaking trials of three treatments (control, MPC1, and MPC2) were conducted. The control cheese (CC) was made from standardized milk (casein-to-fat ratio, C/F approximately 1.7) obtained by mixing skim milk and whole milk (WM); MPC1 and MPC2 cheeses were made from standardized milk (C/F approximately 1.8) obtained from mixing WM and MPC, except that commercial mesophilic starter was added at the rate of 1% to the CC and MPC1 and 2% to MPC2 vats. The addition of MPC doubled cheese yields and had insignificant effects on fat recoveries (approximately 94% in MPC1 and MPC2 vs. approximately 92% in CC) but increased significantly total solids recoveries (approximately 63% in CC vs. 63% in MPC1 and MPC2). Although minor differences were noted in the gross composition of the cheeses, both MPC1 and MPC2 cheeses had lower lactose contents (0.25 or 0.32%, respectively) than in CC (0.60%) 7 d post manufacture. Cheeses from all three treatments had approximately 10(9) cfu/g initial starter bacteria count. The nonstarter lactic acid bacteria (NSLAB) grew slowly in MPC1 and MPC2 cheeses during ripening compared to CC, and at the end of 6 mo of ripening, numbers of NSLAB in the CC were 1 to 2 log cycles higher than in MPC1 and MPC2 cheeses. Primary proteolysis, as noted by water-soluble N contents, was markedly slower in MPC1 and MPC2 cheeses compared to CC. The concentrations of total free amino acids were in decreasing order CC > MPC2 > MPC1 cheeses, suggesting slower secondary proteolysis in the MPC cheeses than in CC. Sensory analysis showed that MPC cheeses had lower brothy and bitter scores than CC. Increasing the amount of starter bacteria improved maturity in MPC cheese.     

Seasonal changes in the fat composition of Lacha sheep's milk used for Idiazabal cheese manufacture

 The effect of the seasonality of lacha sheep's milk production on the milk-fat composition was studied. This raw milk is used to manufacture Idiazabal cheese, a typical product of the Basque Country of northern Spain. Milk samples were taken, prior to cheesemaking, from a refrigerated tank of mixed milk from different flocks in a cheese factory in February, April and June. The triglyceride and fatty acid composition of the milk were analysed. The results showed that the molecular characteristics of the milk-fat components, the degree of unsaturation and the chain length changed with the time of the year. The June milk had a higher content of unsaturated and long-chain fatty acids and triglycerides. The April and February milks were very similar, with a higher content of saturated and short-chain fatty acids and triglycerides. The differences in animal feeding are probably the most important factor affecting milk composition because in June flocks are fed mainly in natural pastures while in April and February the ewes are also fed additional commercial feeds. Idiazabal cheeses were considerably different from February to June, suggesting a relationship between the free fatty acid composition of the cheese, its sensorial properties and the fat composition of the raw milk used in cheesemaking. Received: 18 June 1999     

Influence of microfiltration and adjunct culture on quality of Domiati cheese

The effects of microfiltration and pasteurization processes on proteolysis, lipolysis, and flavor development in Domiati cheese during 2 mo of pickling were studied. Cultures of starter lactic acid bacteria isolated from Egyptian dairy products were evaluated in experimental Domiati cheese for flavor development capabilities. In the first trial, raw skim milk was microfiltered and then the protein:fat ratio was standardized using pasteurized cream. Pasteurized milk with same protein:fat ratio was also used in the second trial. The chemical composition of cheeses seemed to be affected by milk treatment—microfiltration or pasteurization—rather than by the culture types. The moisture content was higher and the pH was lower in pasteurized milk cheeses than in microfiltered milk cheeses at d 1 of manufacture. Chemical composition of experimental cheeses was within the legal limits for Domiati cheese in Egypt. Proteolysis and lipolysis during cheese pickling were lower in microfiltered milk cheeses compared with pasteurized milk cheeses. Highly significant variations in free amino acids, free fatty acids, and sensory evaluation were found among the cultures used in Domiati cheesemaking. The cheese made using adjunct culture containing Lactobacillus delbrueckii ssp. lactis, Lactobacillus paracasei ssp. paracasei, Lactobacillus casei, Lactobacillus plantarum, and Enterococcus faecium received high scores in flavor acceptability. Cheeses made from microfiltered milk received a higher score in body and texture compared with cheeses made from pasteurized milk.     

Characterisation of Macedonian white‐brined cheese: Effect of raw or heat‐treated caprine milk

This research analysed the use of raw and pasteurised milk in the production of Macedonian white caprine milk cheese. Pasteurisation resulted in a considerable decrease in the nitrogen fractions, urea‐polyacrylamide gel electrophoresis of caseins, peptide profiles and volatiles during the maturation period. Forty‐five volatile components were detected, including twelve acids, fourteen esters, six ketones, three alcohols, four terpenes and six other compounds. It was deduced that pasteurisation of milk for the manufacture of artisanal white‐brined cheese delayed the evolution of volatiles, which are related to the unique flavour of the cheese.     

Microbiological and biochemical characteristics of Kashkaval cheese produced using pasteurised or raw milk

Microbiological quality and biochemical changes of Kashkaval cheese manufactured using sheep's raw milk without starter addition or pasteurised milk with an added commercial starter were studied. Mature cheeses had pH values 5.0–5.3, salt content 2.1–2.7%, protein content 23.3–25.1%, moisture content 36.8–39.5%, fat content 28.0–32.2%, and ash content around 5.0%. In raw milk cheeses, mesophilic non-starter lactobacilli prevailed followed by enterococci. In pasteurised milk cheeses Lactococcus lactis starter prevailed. All cheeses were safe according to the criteria in Regulation (EC) 1441/2007. The proteolysis index was around 20%. Butyric, myristic, palmitic, stearic and oleic were the principal free fatty acids in both cheeses. Ketones were abundant in pasteurised milk cheeses and esters in mature raw milk cheeses. Pasteurisation did not affect (P > 0.05) the physicochemical composition and the proteolysis of cheeses. Raw milk cheeses showed higher levels (P < 0.05) of lipolysis than pasteurised milk cheeses.     

Biogenic amine content and proteolysis in Manchego cheese manufactured with Lactobacillus paracasei subsp. paracasei as adjunct and other autochthonous strains as starters

Two different autochthonous strain starter cultures, in which the acidifying starter was composed of strains of Lactococcus lactis, were used for the manufacture of pasteurised milk Manchego cheese. Proteolysis parameters, biogenic amines and sensory characteristics were evaluated and compared with those of commercial starter Manchego cheese and raw milk Manchego cheese manufactured without starter. Autochthonous starter cheeses, and especially those including Lactobacillus paracasei subsp. paracasei as adjunct, presented higher levels of proteolysis than in commercial starter cheese. The concentrations of total biogenic amines in autochthonous starter cheeses were much lower than in raw milk cheese and even lower than in commercial starter cheese. Cheese manufactured with the adjunct strain gave the best results for both flavour intensity and flavour quality, and was the most preferred by panellists. The results suggest that the culture containing Lb. paracasei subsp. paracasei as adjunct could be used for the manufacture of industrial Manchego cheese.     

High Pressure Treatment of Milk and Effects on Microbiological and Sensory Quality of Cheddar Cheese

The effect of cycled high pressure treatment of milk on the yield, sensory, and microbiological quality of Cheddar cheese was investigated. Cheddar cheeses were made from pasteurized, raw, or pressure treated milk according to traditional methods. Flavor scores from trained dairy judges were not different for pasteurized and pressurized milk cheeses (P≤0.05). Percent moisture and wet weight yields of pressure treated milk cheeses were higher than pasteurized or raw milk cheeses (P≤0.05). Microbiological quality of pressurized milk cheeses was comparable to pasteurized milk cheeses. Texture defects were present in pressurized milk cheeses and were attributed to excess moisture. High pressure treatment of milk shows promise as an alternative to heat pasteurization prior to cheesemaking.     

Effect of ripening temperature on the growth and significance of non-starter lactic acid bacteria in Cheddar cheese made from raw or pasteurised milk

Two cheese-making trials were conducted, each involving four cheeses, two made from raw milk (R1, R8) and two from pasteurised milk (P1, P8), and ripened at 1°C (R1, P1) or 8°C (R8, P8). The 1-day-old R1 and R8 cheese in trials 1 and 2 contained ∼10⁴ non-starter lactic acid bacteria (NSLAB) g⁻¹. In trial 1, no NSLAB were detected in 1-day-old P1 and P8 cheeses while those in trial 2 contained 10² cfu g⁻¹. In both trials, the maximum differences between the number of NSLAB in the cheeses ripened at 1 or 8°C were observed at 4 months, when the number of NSLAB in cheeses ripened at 8°C were 3 log cycles higher than in those ripened at 1°C. At the end of ripening (6-months), the number of NSLAB in P8 and R8 were ∼2 log cycles higher than in P1 and R1 cheeses, respectively. Primary proteolysis in the cheeses was markedly affected by ripening temperature, but not by pasteurisation of the cheese milk. Urea-polyacyrlamide gel electrophoretograms and reverse-phase (RP)-HPLC of the water-soluble fraction showed differences between cheeses made from raw or pasteurised milk and between cheeses ripened at 1 or 8°C. The concentration of amino acids and fatty acids were in the order R8>P8>R1>P1. Commercial graders awarded highest flavour scores to the R1 cheeses during gradings at 4, 5 and 6 months. A sensory panel found that most flavour and aroma attributes and maturity were in the order of R8>P8>R1=P1. The results of this study suggest that NSLAB play an important role in the development of flavour in Cheddar cheese by contributing to the production of amino acids and fatty acids.     

Effect of milk pasteurisation temperature on age-related changes in lactose metabolism,pH and the growth of non-starter lactic acid bacteria in half-fat Cheddar cheese

Half-fat Cheddar cheese (∼15%, w/w, fat) was manufactured on three occasions from milk pasteurised at 72, 77, 82 or 87 °C for 26 s, and analysed over a 270 day ripening period. Increasing milk pasteurisation temperature significantly increased the levels of moisture (from ∼45% at 72 °C to 50% at 87 °C), total lactate, and D(−)-lactate in cheese over the 270 day ripening period. Conversely, the cheese pH decreased significantly on increasing pasteurisation temperature. Increasing the pasteurisation temperature did not significantly affect the populations of starter or non-starter lactic acid bacteria during maturation. The use of higher pasteurisation temperatures would appear particularly amenable to exploitation as a means of producing high-moisture (e.g., 40–41%), short-ripened, mild-flavoured Cheddar or Cheddar-like cheeses.