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PubMed2000

A rapid method for the quantitative determination of short-chain free volatile fatty acids from cheese.

The determination of free volatile fatty acids (FVFA) is of interest in the analysis of cheeses. As these compounds are components of taste and flavor, they give indications on metabolic reactions taking place during cheese ripening and can provide an evaluation of cheese defects and their causes. One of the most widely used methods for the determination of FVFA in cheese involves preliminary recovery from the matrix by steam distillation, followed by gas chromatography separation. Relatively high distillate volumes must be collected to achieve a quantitative yield of all the compounds of interest, so that, as a result, the solution is too diluted to achieve good instrumental sensitivity. In this paper, an alternative method for the determination of C2-C6 free carboxylic acids in cheeses involving the use of a Nukol capillary column and crotonic acid as internal standard is described. This method is quick and cheap, as the sample preparation is a simple extraction with water. The underivatized FVFA are then directly separated by gas chromatography. Using this method, all FVFA in cheeses can be quantified with good repeatability and excellent recovery.

Cheese↗
PubMed2001

A large outbreak of Shigella sonnei gastroenteritis associated with consumption of fresh pasteurised milk cheese.

A large outbreak of Shigella sonnei gastroenteritis occurred in Murcia Region (Southeast Spain) in the winter of 1995-1996. More than 200 people were affected. Epidemiological investigations implicated a regionally manufactured fresh pasteurised milk cheese as the vehicle of infection. A case-control study showed a statistically significant association between the illness and consumption of the suspect cheese. The dispersed sale of the cheese resulted in a regional dissemination of the organism and people were affected in eight townships. Research suggested that an infected foodhandler at the cheese factory might have been the source of contamination and that the processing method might have allowed cross-contamination to occur. This study emphasises the importance of increasing the control of strict hygiene during the processing of fresh cheese, since legislation does not forbid direct contact by hand that could result in contamination of cheese even when the milk pasteurisation process was correctly performed.

Adolescent↗
PubMed1997

Inhibition of Listeria monocytogenes by enterocin 4 during the manufacture and ripening of Manchego cheese.

The inhibitory effect of enterocin 4, a bacteriocin produced by Enterococcus faecalis INIA 4, on Listeria monocytogenes strains Ohio and Scott A during the manufacture and ripening of Manchego cheese was investigated. Raw ewe's milk was inoculated with ca 10(5) cfu ml-1 of L. monocytogenes and with 1% of a commercial lactic starter, 1% of an Ent. faecalis INIA 4 culture, or 1% of each culture. Manchego cheeses were manufactured according to usual procedures. Listeria monocytogenes Ohio counts decreased by 3 log units after 8 h and by 6 log units after 7 d in cheese made from milk inoculated with Ent. faecalis INIA 4 or with both cultures, whereas no inhibition was recorded after 60 d in cheese made from milk inoculated with commercial lactic starter. Listeria monocytogenes Scott A was not inhibited by enterocin 4 during cheese manufacture, but decreases of 1 log unit after 7 d and of 2 log units after 60 d were achieved in cheese made from milk inoculated with both commercial lactic starter and Ent. faecalis INIA 4.

Bacteriocins↗
PubMed1997

Inhibition of Listeria monocytogenes by piscicolin 126 in milk and Camembert cheese manufactured with a thermophilic starter.

The effect of bacteriocin, piscicolin 126, on the growth of Listeria monocytogenes and cheese starter bacteria was investigated in milk and in Camembert cheese manufactured from milk challenged with 10(2) cfu ml(-1) L. monocytogenes. In milk incubated at 30 degrees C, piscicolin 126 added in the range of 512-2,048 AU ml(-1) effectively inhibited growth of L. monocytogenes for more than 20 d when challenged with approximately 10(2) cfu ml(-1) L. monocytogenes. At higher challenge levels (10(4) and 10(6) cfu ml(-1)), piscicolin 126 reduced the viable count of L. monocytogenes by 4-5 log units immediately after addition of the bacteriocin; however, growth of Listeria occurred within 24 h. The minimum inhibitory concentration (MIC) of piscicolin 126 against lactic acid cheese starter bacteria was generally greater than 204,800 AU ml(-1) , and the viable count and acid production of these starter cultures in milk were not affected by the addition of 2,048 AU ml(-1) piscicolin 126. Camembert cheeses made from milk challenged with L. monocytogenes and with added piscicolin 126 showed a viable count of L. monocytogenes 3-4 log units lower than those without piscicolin 126. Inactivation of piscicolin 126 by proteolytic enzymes from cheese starter bacteria and mould together with the emergence of piscicolin 126-resistant isolates was responsible for the recovery of L. monocytogenes in the cheeses during ripening.

Animals↗
PubMed2002

Genetic diversity of lactococci and enterococci isolated from home-made Pecorino Sardo ewes' milk cheese.

AIMS: To assess the intraspecific genetic diversity of lactococci and enterococci isolated from 24-h, 1- and 2-month-old home-made Pecorino Sardo ewes' milk cheese. METHODS AND RESULTS: Two molecular techniques, plasmid profiling and pulsed-field gel electrophoresis, were used in order to type the isolates at strain level. The present study revealed that the lactococcal and enterococcal microbial populations of home-made Pecorino Sardo cheese were complex, not only 24 h after manufacture, but also after 1 and 2 months of ripening. The genetic diversity at subspecies level ranged from 58 to 80% during the three periods examined. The study also showed that the strains that dominated in the first stage of ripening were not necessarily predominant in the later periods. A high number of strains isolated at 24 h were still present in the mature cheese, but many of the genotypes were only found in the cheese after 1 or 2 months. CONCLUSIONS: The results showed a high intraspecific genetic diversity in the natural microbial population colonizing home-made Pecorino Sardo cheese. Two molecular techniques are necessary for a thorough and precise typing at strain level in order to better distinguish between closely related isolates and between isolates that probably belong to the same clonal lineage. SIGNIFICANCE AND IMPACT OF THE STUDY: The genetic complexity observed in the present study is of particular relevance in the preservation of the natural microflora of traditional Protected Designation of Origin raw milk cheeses, as well as in the selection of new starter strains for the dairy industry.

Animals↗
PubMed2002

Occurrence of Staphylococcus and enteropathogens in soft cheese commercialized in the city of Rio de Janeiro, Brazil.

AIMS: To investigate the presence of Staphylococcus aureus, enteropathogenic Escherichia coli (EPEC), Aeromonas spp. and Yersinia spp. in soft cheese commercialized in Rio de Janeiro, Brazil. METHODS AND RESULTS: A total of 45 samples of cheese from three different brands marketed in Rio de Janeiro city were analysed for faecal coliform levels using the Most Probable Number (MPN) technique. The samples were also analysed using conventional methodology for the investigation of food-borne pathogens. High levels of faecal contamination were detected in 95.5% of cheese samples. Staphylococcus aureus was isolated from 20% of samples, of which 17.7% were above the limits allowed by Brazilian legislation. Aeromonas hydrophila and Aer. caviae were detected in 17.7% of the samples. Yersinia spp. were not found in this study. EPEC was isolated from 21.1% of the samples and the most frequently found serogroups were O127, followed by O55 and O26. CONCLUSIONS: Our results showed that 95.5% of cheese samples had high levels of faecal coliforms. The isolation of Staph. aureus, serogroups of EPEC and Aeromonas spp. suggested that the soft cheese commercialized in the city of Rio de Janeiro may represent a health risk for the consumers. SIGNIFICANCE AND IMPACT OF THE STUDY: These results suggest that soft cheese may act as an important vehicle of transmission for well-established pathogens.

Aeromonas↗
PubMed2003

Evaluation of culture media for the recovery of Mycobacterium avium subsp. paratuberculosis from Cheddar cheese.

AIMS: The study evaluated the efficacy of four Mycobacterium avium subsp. paratuberculosis (MAP) culture media in suppressing commonly used starter cultures and typical nonstarter microflora present during the manufacture and ripening of Cheddar cheese, with a view to identify a suitable medium for the enumeration of MAP during laboratory-scale Cheddar production. METHODS AND RESULTS: Four Cheddar starter cultures and Cheddar cheese manufactured with these starters were inoculated onto Herrold's egg yolk medium (HEYM); HEYM supplemented with vancomycin, amphotericin B and nalidixic acid (HEYM/VAN); Middlebrook 7H10 agar containing polymyxin, amphotericin B, nalidixic acid, trimethoprim and azlocillin (PANTA) antibiotic supplement; and BACTEC 12B radiometric medium with and without a preliminary decontamination step (0.75% w/v hexadecylpyridinium chloride (HPC), 5 h). The inclusion of a decontamination step inhibited all Cheddar cheese starter and nonstarter micro-organisms. The medium 7H10/PANTA and to a lesser extent HEYM/VAN were effective inhibitors of cheese microflora when no decontamination step was employed. CONCLUSIONS: Middlebrook 7H10 medium, supplemented with PANTA antibiotics, suppressed all micro-organisms associated with ripening Cheddar cheese manufactured with pasteurized milk. SIGNIFICANCE AND IMPACT OF THE STUDY: A MAP culture medium has been identified, which may be used to enumerate this bacterium during the laboratory manufacture and ripening of Cheddar cheese and hence facilitate further research into the persistence of this pathogen in the product.

Animals↗
PubMed2003

Caseinolysis in cheese by Enterobacteriaceae strains of dairy origin.

AIMS: To study the effect of Enterobacteriaceae strains of dairy origin on caseins under cheese manufacture and ripening conditions. METHODS AND RESULTS: Strains belonging to the genera Enterobacter, Escherichia, Hafnia and Serratia were isolated from fresh raw milk cheeses. Residual caseins in cheeses made from milk individually inoculated with 10 strains of Enterobacteriaceae were determined by capillary electrophoresis. Hierarchical cluster analysis of strains based on data of residual caseins grouped together strains from the same genus, excepting Hafnia strains, which were separated into two groups. Serratia was the most proteolytic genus in our study. Preferences for degradation of casein fractions differed among the four genera studied. CONCLUSIONS: Enterobacteriaceae strains posses proteolytic systems active on all casein fractions under cheese manufacture and ripening conditions. The effects on caseins were similar for strains belonging to the same genus. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of Enterobacteriaceae in cheeses may affect proteolysis during ripening. Assays of Enterobacteriaceae proteolytic activity on milk agar plates may underestimate their caseinolytic activity in cheese.

Animals↗
PubMed2000

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).

Cheese↗
PubMed2006

Distribution and stability of Aflatoxin M1 during processing and ripening of traditional white pickled cheese.

The distribution of aflatoxin M(1) (AFM(1)) has been studied between curd, whey, cheese and pickle samples of Turkish white pickled cheese produced according to traditional techniques and its stability studied during the ripening period. Cheeses were produced in three cheese-making trials using raw milk that was artificially contaminated with AFM(1) at the levels of 50, 250 and 750 ng/l and allowed to ripen for three months. AFM(1) determinations were carried out at intervals by LC with fluorescence detection after immunoaffinity column clean-up. During the syneresis of the cheese a proportionately high concentration of AFM(1) remained in curd and for each trial the level was 3.6, 3.8 and 4.0 times higher than levels in milk. At the end of the ripening, the distribution of AFM(1) for cheese/whey + brine samples was 0.9, 1.0 and 1.3 for first, second and third spiking respectively indicating that nearly half of the AFM(1) remained in cheese. It has been found that only 2-4% of the initial spiking of AFM(1) transferred into the brine solution. During the ripening period AFM(1) levels remained constant suggesting that AFM(1) was quite stable during manufacturing and ripening.

Aflatoxin M1↗
PubMedDate not supplied

Migration of mineral hydrocarbons into foods. 3. Cheese coatings and temporary casings for skinless sausages.

Levels of mineral hydrocarbons which have migrated from wax coatings into cheese have been determined for 20 retail samples using a gas chromatographic procedure. Contamination was limited to the outermost 2 mm of cheese in direct contact with the wax where levels of hydrocarbons were found to range from 10 to 150 mg/kg. On a whole cheese weight basis these amounted to < 1 to 27 mg/kg (< 0.2 to 3 mg/dm2 contact area). Components attributed to hydrocarbons in cheese samples remote from the waxed surface (background levels) were typically 3-5 mg/kg. Background levels were subtracted from the results for surface samples to obtain migration values. There was evidence that the surface contamination of cheese with mineral hydrocarbons occurred by a combination of diffusion into the cheese and adhesion of wax components onto its surface. Mineral hydrocarbons are used in the manufacture of the temporary casings used to mould skinless sausages. Of 33 retail products examined, including skinless sausages, hot-dog sausages and frankfurters, 25 (75%) contained levels of mineral hydrocarbons from 10 to 105 mg/kg. These hydrocarbons were shown to be present principally at the surface of the food and so could be attributed to migration. Nine other minced meat products were examined for comparison, including minced beef, pâté, sausage meat and sausages with skins. Levels of mineral oil in these products were insignificant by comparison, typically below the limit of detection of ca 4 mg/kg, indicating insignificant adventitious contamination from routes other than migration.

Animals↗
PubMed2004

Survival of Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum BB-02 in white-brined cheese.

The viability of probiotic cultures used as adjunct cultures including Bifidobacterium bifidum BB-02 and Lactobacillus acidophilus LA-5 in white-brined cheese was investigated. The microbiological and biochemical properties of cheeses were assessed throughout 90 days of storage. Two inoculum rates for probiotic starters (2.5% and 5.0%, v/v, corresponding to (1.0-1.3)x10(9) cfu/g and (2.0-2.1)x10(9) cfu/g, respectively) were compared with the control cheese. Results obtained showed that a higher inoculum rate resulted in faster proteolysis. The cheese inoculated with 5% probiotic strains had higher water-soluble nitrogen, non-protein nitrogen, proteose-peptone nitrogen and tyrosine values than the other experimental cheeses. At the end of storage, the cheese supplemented with 5% probiotic culture had a ripening coefficient of 28.3%, followed by 2.5% probiotic strains containing the sample being 24.9%, and the control being 23.6%, respectively. It was also found that the colony counts of both probiotic microorganisms declined during storage. After 90 days of storage, the number of probiotic colonies were still around the threshold for a minimum probiotic effect (10(6) cfu/g).

Bifidobacterium↗
PubMed2005

Chemical characteristics, fatty acid compositions, conjugated linoleic acid contents and cholesterol levels of some traditional Turkish cheeses.

The chemical characteristics, fatty acid and conjugated linoleic acid (CLA) contents and cholesterol levels of some traditional Turkish cheeses that are consumed mostly in Turkey were determined in this study. The fatty acid and cholesterol contents and CLA amount of cheeses were analysed as methyl esters by gas chromatography. The aim of this study was to determine the nutritional profile of some Turkish cheeses produced by traditional methods. The major fatty acids of the cheeses were palmitic acid (C(16:0)) and oleic acid (C(18:1)). The saturated fatty acid content of samples changed between 60.80% and 76.57%, while the monounsaturated fatty acid content ranged from 21.42% to 34.05% and the polyunsaturated fatty acid content was between 1.47% and 3.59%. The CLA contents of the cheeses ranged from 0.44 to 1.04 g/100 g in fat. The cholesterol levels of the samples were determined as 44.6-147.69 mg/100 g in cheese.

Cheese↗
PubMed2003

Use of butter and cheese in 10 European countries: a case of contrasting educational differences.

BACKGROUND: This paper aims to analyse socioeconomic variation in the use of cheese and butter in Europe by reviewing existing dietary surveys. It explores whether socioeconomic differences in the intake of these foods follow a similar pattern in all countries. METHODS: An overview of available studies on socioeconomic differences in food habits in Europe over the period 1985-1997 was performed. Twenty studies from 10 countries included information on cheese and butter. A simple directional vote-counting method was used to register the association between educational level and consumption of cheese and butter (animal fat) for each study. FAO's food balance sheets were used to classify the countries according to consumption trends of these foodstuffs. RESULTS: In all countries higher social classes used more cheese than lower classes. The results for butter were less consistent. In the Nordic countries higher social classes used less butter than lower ones. In the other countries an opposite pattern or no differences could be observed. However, in countries where the use of both butter and animal fats could be analysed, animal fats were used more by the lower social classes. CONCLUSIONS: Higher and lower socioeconomic groups have different sources of saturated fats. Higher social classes use more cheese whereas lower social classes use more butter or animal fats. This can be observed especially in countries where the consumption of cheese is increasing and that of butter decreasing. Higher social classes prefer modern foods, lower classes traditional foods.

Butter↗
PubMed1993

Study of Enterobacteriaceae throughout the manufacturing and ripening of hard goats' cheese.

The evolution of the counts and the species of Enterobacteriaceae as well as some physico-chemical parameters (pH, aw and NaCl and moisture contents) during manufacturing and ripening of a hard Spanish goats' cheese of the Armada-Sobado variety were studied. Enterobacteriaceae (mean log counts 4.45 g-1 in milk) increased 0.71-2.18 log units in curd and afterwards decreased until they disappeared after 2-4 weeks of ripening. This premature disappearance seems to be due to the decrease in aw values and in moisture contents. However, the low pH values, reached from the beginning of the ripening process, could also contribute to this phenomenon. The most abundant species in milk was Serratia liquefaciens (57.5% of isolates), followed by Morganella morganii (27.5%), Hafnia alvei (5%), Klebsiella oxytoca (5%) and Yersinia enterocolitica (5%). Yersinia enterocolitica was not subsequently isolated from either curd or in cheese. Hafnia alvei numbers increased in curd and in 1-week-old cheese where this micro-organism was the most abundant (47.5% and 75% of the isolates respectively). Escherichia coli, which was not isolated from milk, curd or 1-week-old cheese, was the predominant organism in 2-week-old cheese (57.8% of isolates). This confirms the finding of other authors who have shown that it is one of the most resistant species in ripening cheeses.

Animals↗
PubMed2005

Cheddar cheese cooking temperature induces differential lactococcal cell permeabilization and autolytic responses as detected by flow cytometry: implications for intracellular enzyme accessibility.

AIMS: To determine the influence of cheese cooking temperature on autolysis and permeabilization of two lactococcal starter strains in broth and in Cheddar cheese juice during ripening. METHODS AND RESULTS: Flow cytometry (FCM) was used to identify and enumerate intact and permeabilized cells in broth and in Cheddar cheese juice. Levels of intracellular enzyme activities were quantified concurrently. Permeabilized cell numbers increased for both strains in broth following a temperature shift from 32 to 38 degrees C and was accompanied by an increase in the level of accessible intracellular enzyme activities. The relative proportions of intact and permeabilized cell populations, as detected by FCM in cheese juice, changed during 42-day ripening. Permeabilized cell populations increased during ripening for both strains; however, an increase in accessible intracellular enzyme activity was observed only for the highly autolytic strain Lactococcus lactis AM2. CONCLUSIONS: Differences in the autolytic and permeabilization response induced by cooking temperature in two lactococcal strains affects intracellular enzyme accessibility in Cheddar cheese. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the importance of the autolytic and permeabilization properties of lactic acid bacteria starter strains and their impact on cheese ripening.

Bacteriolysis↗
PubMed2006

Proteolytic enzyme activities in Cheddar cheese juice made using lactococcal starters of differing autolytic properties.

AIMS: To determine proteolytic enzyme activities released in Cheddar cheese juice manufactured using lactococcal starter strains of differing autolytic properties. METHODS AND RESULTS: The activities of residual chymosin, cell envelope proteinase and a range of intracellular proteolytic enzymes were determined during the first 70 days of ripening when starter lactococci predominate the microbial flora. In general, in cell free extracts (CFE) of the strains, the majority of proteolytic activities was highest for Lactococcus lactis HP, intermediate for L. lactis AM2 and lowest for L. lactis 303. However, in cheese juice, as ripening progressed, released proteolytic activities were highest for the highly autolytic strain L. lactis AM2, intermediate for L. lactis 303 and lowest for L. lactis HP. CONCLUSIONS: These results indicate that strain related differences in autolysis influence proteolytic enzyme activities released into Cheddar cheese during ripening. No correlation was found between proteolytic potential of the starter strains measured in CFE prior to cheese manufacture and levels of activities released in cheese juice. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings further support the importance of autolysis of lactococcal starters in determining the levels of proteolytic activities present in cheese during initial stages of ripening.

Cheese↗
PubMed2005

Design and application of a Staphylococcus-specific single strand conformation polymorphism-PCR analysis to monitor Staphylococcus populations diversity and dynamics during production of raw milk cheese.

AIM: Development of a nested-PCR single strand conformation polymorphism (SSCP) assay targeting the 16S rRNA genes of the Staphylococcus genus, to monitor staphylococci in cheese. METHODS AND RESULTS: New primer sets to specifically amplify 16S rDNA of staphylococci were designed to be used in a nested-PCR SSCP assay. The method was efficient in discriminating the staphylococcal species most frequently found in cheese. It was validated by monitoring Staphylococcus populations in three productions of raw milk cheese. Analysis of milk samples revealed dominant SSCP peaks corresponding to Staphylococcus aureus, Staphylococcus equorum and Staphylococcus saprophyticus. After 12 h, the S. aureus peak became dominant. CONCLUSIONS: The combination of specific Staphylococcus nested-PCR and SSCP allows rapid and direct monitoring of staphylococci diversity and dynamics in milk and cheese. In the core of the cheeses studied, S. aureus may have ecological advantages against other Staphylococcus populations. SIGNIFICANCE AND IMPACT OF THE STUDY: This approach is a promising tool to study the ecology of staphylococci in cheeses and in other food samples.

Animals↗