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Survival of surface ripening cultures during storage and monitoring their development on cheese.

AIMS: To study the survival of bacteria isolated from the surface of smear cheese and monitor their development during cheese ripening. METHODS AND RESULTS: The storage of five potential bacterial surface-ripening cheese cultures, Brevibacterium aurantiacum, Corynebacterium casei, Corynebacterium variable, Microbacterium gubbeenense and Staphylococcus saprophyticus, in maximum recovery diluent (MRD), containing 0.85% w/v or 5% w/v NaCl, at 21 or 4 degrees C for 40 days, was investigated. All five strains studied survived well with a maximum decrease of c. 2.5 log(10) CFU ml(-1) after storage for 40 days at 4 degrees C in 0.85% or 5% w/v NaCl. Survival, especially of C. variable, was less at 21 degrees C. The development of defined ripening cultures containing C. casei and Debaryomyces hansenii on two farmhouse cheeses was also evaluated. Using pulsed-field gel electrophoresis (PFGE) for the bacteria and mitochondrial DNA restriction fragment length polymorphism (mtDNA-RFLP) for the yeast, it was shown that the ripening cultures could be re-isolated in high numbers, 10(8) CFU cm(-2) for C. casei and 10(6) CFU cm(-2) for D. hansenii, from the cheese surface after 2.5 weeks of ripening. CONCLUSIONS: Ripening strains of surface ripening cultures can be stored in MRD containing 5% w/v salt at 4 degrees C for at least 40 days. Such cultures are recovered in high numbers from the cheese during ripening. SIGNIFICANCE AND IMPACT OF STUDY: This study has provided a low-cost and efficient way to store bacteria that could be used as ripening cultures for smear cheese. Such cultures can be recovered in high numbers from the cheese surface during ripening.

Bacteria↗

Occurrence of Streptococcus macedonicus in Italian cheeses.

A new approach for the detection and enumeration of Streptococcus macedonicus in cheese was developed. The method which is based on a first screening of cheeses by a PCR assay specific for S. macedonicus followed by plating positive samples on a differential medium (SM medium) was applied to 51 samples derived from PDO and traditional Italian cheeses. Streptococcus macedonicus was found in 16 of the 51 samples examined in the present work. With the exclusion of an Asiago cheese sample in which very high numbers of S. macedonicus (7.13 log CFU g(-1)) were found, the counts of S. macedonicus in SM medium ranged from 2.48 to 4.70 log CFU g(-1). In the same cheeses, total streptococci enumerated onto M17 agar were found at higher concentrations with values up to 7.88 log CFU g(-1). The system developed was particularly useful for the differential count of S. macedonicus in cheese and allowed to evaluate the occurrence of this species within the complex microbial lactic acid bacteria (LAB) population, which is typical of traditional cheeses. Results showed that in the examined cheeses S. macedonicus cannot be considered as a dominant LAB species.

Cheese↗

Methods for improved recovery of Listeria monocytogenes from cheese.

Method of homogenization (Waring blender versus stomacher), type of diluent (tryptose broth [TB] versus aqueous 2% trisodium citrate), and temperature of diluent (20 versus 40 degrees C) were compared for recovery of Listeria monocytogenes from freshly made and ripened Colby cheese. By using direct plating on McBride listeria agar, significantly higher numbers of L. monocytogenes were recovered when cheese samples were (i) homogenized for 2 min with the blender rather than the stomacher (P less than 0.01), (ii) diluted in trisodium citrate rather than TB (P less than 0.01), and (iii) diluted in diluents at 40 rather than 20 degrees C (P less than 0.05). Based on these results, a new diluent/enrichment medium was developed by adding 2% trisodium citrate to TB (TBC). Despite superior results with the blender, biosafety concerns led to use of the stomacher for homogenization of cheese samples; hence, the stomaching time was increased to 3 min. Results obtained by direct plating indicated that recovery of L. monocytogenes from Colby cheese and from curd samples taken during manufacture of brick cheese increased when samples were diluted 1:10 in TBC at 45 degrees C and stomached for 3 min, as compared with similarly treated samples diluted in TB at 25 degrees C. A similar comparison of both diluents for recovery of L. monocytogenes from cold-pack cheese food yielded bacterial counts which were not significantly different. Recovery of L. monocytogenes from cold-enriched (at 4 degrees C for up to 8 weeks) samples of Colby cheese and cold-pack cheese food was generally similar for samples homogenized in TBC or TB.

Animals↗

Contribution of Lactococcus lactis cell envelope proteinase specificity to peptide accumulation and bitterness in reduced-fat Cheddar cheese.

Bitterness is a flavor defect in Cheddar cheese that limits consumer acceptance, and specificity of the Lactococcus lactis extracellular proteinase (lactocepin) is widely believed to be a key factor in the development of bitter cheese. To better define the contribution of this enzyme to bitterness, we investigated peptide accumulation and bitterness in 50% reduced-fat Cheddar cheese manufactured with single isogenic strains of Lactococcus lactis as the only starter. Four isogens were developed for the study; one was lactocepin negative, and the others produced a lactocepin with group a, e, or h specificity. Analysis of cheese aqueous extracts by reversed-phase high-pressure liquid chromatography confirmed that accumulation of alpha(S1)-casein (f 1-23)-derived peptides f 1-9, f 1-13, f 1-16, and f 1-17 in cheese was directly influenced by lactocepin specificity. Trained sensory panelists demonstrated that Cheddar cheese made with isogenic starters that produced group a, e, or h lactocepin was significantly more bitter than cheese made with a proteinase-negative isogen and that propensity for bitterness was highest in cells that produced group h lactocepin. These results confirm the role of starter proteinase in bitterness and suggest that the propensity of some industrial strains for production of the bitter flavor defect in cheese could be altered by proteinase gene exchange or gene replacement.

Cheese↗

Source of enterococci in a farmhouse raw-milk cheese.

Enterococci are widely distributed in raw-milk cheeses and are generally thought to positively affect flavor development. Their natural habitats are the human and animal intestinal tracts, but they are also found in soil, on plants, and in the intestines of insects and birds. The source of enterococci in raw-milk cheese is unknown. In the present study, an epidemiological approach with pulsed-field gel electrophoresis (PFGE) was used to type 646 Enterococcus strains which were isolated from a Cheddar-type cheese, the milk it was made from, the feces of cows and humans associated with the cheese-making unit, and the environment, including the milking equipment, the water used on the farm, and the cows' teats. Nine different PFGE patterns, three of Enterococcus casseliflavus, five of Enterococcus faecalis, and one of Enterococcus durans, were found. The same three clones, one of E. faecalis and two of E. casseliflavus, dominated almost all of the milk, cheese, and human fecal samples. The two E. casseliflavus clones were also found in the bulk tank and the milking machine even after chlorination, suggesting that a niche where enterococci could grow was present and that contamination with enterococci begins with the milking equipment. It is likely but unproven that the enterococci present in the human feces are due to consumption of the cheese. Cow feces were not considered the source of enterococci in the cheese, as Enterococcus faecium and Streptococcus bovis, which largely dominated the cows' intestinal tracts, were not found in either the milk or the cheese.

Animals↗

Effect of raw-milk cheese consumption on the enterococcal flora of human feces.

Enterococci are one of the major facultative anaerobic bacterial groups that reside in the human gastrointestinal tract. In the present study, the composition of the enterococcal fecal flora in three healthy humans was analyzed before, during, and after the daily consumption of approximately 125 g of a raw-milk Cheddar-type cheese containing 3.2 x 10(4) enterococci/g of cheese. Enterococcal counts ranged between 1.4 x 10(2) and 2.5 x 10(8) CFU/g of feces and differed from subject to subject and from week to week. The cheese contained mainly Enterococcus casseliflavus and a small population of Enterococcus faecalis. Clonal relationships were determined by pulsed-field gel electrophoresis. Before and after consumption of the cheese, samples from humans contained mainly Enterococcus faecium, with some of the clones being resident. During consumption of the cheese, one particular transient clone of E. faecalis, clone Fs2, which was present in small numbers in the cheese, largely dominated the feces. Two clones of E. casseliflavus from the cheese were also found in the feces of one of the subjects during cheese consumption. These results suggest that a clone need not be present in a food in high numbers to establish itself in the intestine.

Animals↗

Storage stability of Clostridium botulinum toxin and spores in processed cheese.

Growth initiated from detoxified spores of Clostridium botulinum 62A resulted in toxin production of 50 to 10,000 mouse lethal doses (MLD) per gram of processed soft surface-ripened cheese. Regular assays during subsequent storage of toxic samples at 2 to 4 C revealed a characteristic two- to fivefold increase in toxin titer during the initial 1 week to 12 months of storage. Thereafter, the toxin titer remained constant for 2 to 4 years, after which the toxicity declined rapidly. At the end of 6 years of storage at 2 to 4 C, the samples still contained 20 to 5,000 MLD of toxin per gram, with the usual toxin level at 200 to 500 MLD. Toxic culture filtrates of C. botulinum incorporated into cheese and stored at 30 C for 60 days showed no decline in toxin in processed type I cheese, but toxin decreased slightly in processed type II and type III cheese. The surface flora of these cheeses did not attack but, on the contrary, protected C. botulinum toxin during storage at 30 C. Initial difficulties in recovering C. botulinum organisms from type I cheese were traced to growth inhibitory activity which could be removed by washing with distilled water in a centrifuge. Viable spores or vegetative cells could be recovered from all samples after 4 to 5 years of storage at 2 to 4 C. After 6 years, organisms were recovered from all except three samples of type I cheese. Two other samples showed a large decrease in viable organisms. In type III cheese, spores remained remarkably stable for 6 years at the level of the initial inoculum, i.e., approximately 10(5) spores per gram.

Cheese↗

Effect of eating cheese on Ca and P concentrations of whole mouth saliva and plaque.

A study was undertaken to examine the release of calcium and phosphate from cheese during mastication. Unstimulated saliva was collected for baseline analysis in the initial study followed by saliva collection after chewing different cheeses with and without biscuits. In the second study, volunteers who had abstained from tooth cleaning for 24 h had plaque samples taken from two quadrants, they then chewed cheese in their own personal eating manner, and a second sample of plaque was taken within 5 min. The results showed that the calcium ion concentration of the oral fluids rose from a mean of 30 micrograms/ml to between 200 and 540 micrograms/ml, depending on the type of cheese, but the phosphate concentration fell below baseline. The release of both ions tended to be less when the cheese was eaten with a biscuit. In the second study a highly significant rise in plaque calcium concentration was shown after eating cheese, but no consistent change in phosphate level was found. Acidic soft drinks, following eating, tended to reduce the plaque calcium levels, but no consistent change was found if tea or coffee was taken following the cheese consumption. It is suggested, from these findings, that cheese eaten alone at the very end of a meal raises plaque calcium and might be effective in reducing dental caries.

Beverages↗

Effect of cheese, with and without sucrose, on dental caries and recovery of Streptococcus mutans in rats.

The objective of this study was to determine the effect of aged and young cheddar cheese with and without added sucrose on dental caries and the associated recovery of implanted Streptococcus mutans. Very little caries was observed in rats consuming cheese without sucrose. There was an increase in caries in rats fed cheeses with 20% sucrose, but this increase was not significant. There was significantly greater caries activity in rats fed standard diets containing 20% or 5% sucrose (SLS or MIT 305) than in rats fed cheeses containing 20% sucrose. Rats fed cheese or powdered diets containing sucrose had significantly higher frequency of recovery and higher levels of S. mutans infection than did rats fed cheese containing no sucrose. This study confirms the low cariogenic potential and possible cariostatic activity of cheddar cheese in rats. Since cheddar cheese with sucrose did not significantly interfere with S. mutans implantation, the cariostatic mechanism is apparently unrelated to a direct antimicrobial effect on S. mutans.

Animals↗

[Microscopic analysis of "prato", "mussarela" and "mineiro" cheese sold in street markets of the City of São Paulo, southeastern Brazil].

INTRODUCTION: Cheese should be produced from ingredients of good quality and processed under hygienic conditions. Further, cheese should be transported, stored and sold in an appropriate manner in order to avoid, among other things, the incorporation of extraneous materials (filth) of biological origin or otherwise, in contravention of the relevant food legislation. The aim of the study was to evaluate the hygienic conditions of "prato", "mussarela", and "mineiro" cheeses sold at the street food markets in the city of S. Paulo, Brazil. MATERIALS AND METHOD: Forty-seven samples of each of the three types of cheese were collected during the period from March, 1993 to February, 1994. The Latin square was used as a statistical model for sampling and random selection of the street markets from which to collect the cheese samples. The samples were analysed for the presence of extraneous matters outside for which purpose the samples were washed and filtered and inside, for which the methodology of enzymathic digestion of the sample with pancreatine, followed by filtering, was used. RESULTS AND CONCLUSIONS: Of the 141 samples analysed, 75.9% exhibited at least one sort of extraneous matters. For the "prato" and "mussarela" cheeses, the high number of contaminated samples was due mainly to extraneous matters present inside the cheese, whereas in the "mineiro" cheese, besides the internal filth, 100% of the samples had external filth.

Brazil↗

Influence of the season on proteolysis and sensory characteristics of Idiazabal cheese.

The present study examined the influence of the manufacturing time on proteolysis and sensory characteristics of Idiazabal cheese. Samples of Idiazabal cheeses made using traditional manufacturing methods at two different times of year were collected at a plant operating under the auspices of the Idiazabal Cheese Appellation of Origin. The cheeses were made in February and June, roughly the beginning and the end of the manufacturing season. Chemical analyses were performed on d 1, 90, and 180 of ripening and sensory analyses on d 90, 120, and 180 of ripening. Higher nitrogen fraction (soluble and nonprotein nitrogen) levels and free amino acid contents were observed in the cheeses manufactured during the month of June. Degradation of the alphas- and beta-caseins was also greater in the cheeses made at that time of year. The cheeses manufactured in February earned higher sensory analysis scores for characteristic odor and taste and higher total sensory scores. The cheeses manufactured in June earned higher scores for the sensory attributes sweet flavor and bitter taste.

Animals↗

Effect of pasteurization of Ewe's milk and use of a native starter culture on the volatile components and sensory characteristics of roncal cheese.

The effect of pasteurization of milk and use of a native starter culture on the volatile components and sensory characteristics of a Spanish ewe's-milk cheese were examined. Three cheese batches were made, one from raw milk, another from pasteurized milk, and a third from pasteurized milk with an added native starter culture in addition to the commercial starter. Cheeses were analyzed at 1, 120, and 240 d of ripening. Analysis of the volatile components was by purge and trap connected to a gas chromatograph with a mass spectrometer and disclosed a total of 76 components belonging to the following chemical families: hydrocarbons, fatty acids, esters, sulfur and carbonyl compounds, and, in particular, alcohols. Pasteurization lowered the levels of certain volatile components, especially alcohols, aldehydes, and ketones. The cheeses made from pasteurized milk showed lower scores for attributes of characteristic taste and aftertaste, as well as a characteristic aroma at 240 d of ripening. These results suggest that the components present in higher concentrations in the cheeses made from the raw milk were necessary for development of characteristic Roncal cheese aroma. The new native starter culture tested did not exert a significant effect on any of the parameters considered, with the exception of certain isolated components, for which higher or lower quantities were recorded in the cheeses made with that starter culture, although the differences did not have a definite effect on the sensory characteristics of the cheeses.

Animals↗

Effect of milk preacidification on low fat mozzarella cheese: II. Chemical and functional properties during storage.

The effect of milk preacidification on cheese manufacturing, chemical properties, and functional properties of low fat Mozzarella cheese was determined. Four vats of cheese were made in 1 d using no preacidification (control), preacidification to pH 6.0 and pH 5.8 with acetic acid, and preacidification to pH 5.8 with citric acid. This process was replicated four times. Modifications in the typical Mozzarella manufacturing procedures were necessary to accommodate milk preacidification. The chemical composition of the cheeses was similar among the treatments, except the calcium content and calcium as a percentage of protein were lower in the preacidified treatments. During refrigerated storage, the chemical and functional properties of low fat Mozzarella were affected the most by milk preacidification to pH 5.8 with citric acid. The amount of expressible serum, unmelted cheese whiteness, initial unmelted hardness, and initial apparent viscosity were lower with preacidification. The reduction in initial unmelted cheese hardness and initial apparent viscosity in the pH 5.8 citric treatments represents an improvement in the quality of low fat Mozzarella cheese that allows the cheese to have better pizza bake characteristics with shorter time of refrigerated storage.

Acetic Acid↗

Microbiological and biochemical properties of canestrato pugliese hard cheese supplemented with bifidobacteria.

Canestrato Pugliese cheeses from ewe milk were produced according to a traditional protocol and by adding 7.0 log10 cfu of fresh cells per gram of Bifidobacterium bifidum Bb02, Bifidobacterium longum Bb46, or both species. The traditional technology was modified slightly to favor the survival of probiotic microorganisms. After 56 d of ripening, the survival of B. bifidum Bb02 and B. longum Bb46 was 6.0 and 5.0 log10 cfu/g, respectively. After 19 d cheeses contained ca. 7.0 log10 cfu/g of bifidobacteria. Compared to traditional cheese, the addition of bifidobacteria seemed to support the growth and survival of mesophilic lactobacilli and Streptococcus thermophilus, used as starter, during ripening. No significant differences were observed in the main chemical composition, and only a slightly higher concentration of acetic acid was found in cheeses with bifidobacteria added. On the contrary, alpha- and beta-galactosidase activities were markedly more pronounced in the presence of bifidobacteria, especially with B. bifidum Bb02. In contrast with traditional cheese, the lactose was completely hydrolyzed in cheeses made with bifidobacteria. Urea-PAGE electrophoresis of the pH 4.6-soluble and pH 4.6-insoluble N fractions did not show appreciable variations. Only the reversed-phase-HPLC analysis of the pH 4.6-soluble N showed a slightly more complex profile in the presence of bifidobacteria. This finding was in agreement with the higher value of the pH 4.6-soluble N/total N ratio and with the more pronounced amino-, imino-, and dipeptidase activities found in all the cheeses with the bifidobacteria added, especially B. bifidum Bb02. No differences were found in the free amino acid and free fatty acid contents. The amino acids glutamic acid, valine, proline, leucine, and lysine and the fatty acids butyric, caproic, capric, and oleic acids were found at the highest concentrations. The sensory evaluation did not show significant differences, and Canestrato Pugliese cheeses were characterized by small and uniformly distributed eyes, were pale yellow, had an elastic consistency and a Pecorino-like smell, were very salty, and tended to be moderately piquant.

Amino Acids↗

Effect of heat treatments on the meltability of cheeses.

The effect of heat treatments on the meltability of cheese was investigated. Cheddar cheeses of different composition and low-moisture, part-skim Mozzarella cheese were studied at 1, 3, 6, and 12 wk of aging. Cheese samples were heated to 60 degrees C and held for 0, 10, and 20 min before allowing the melted cheese to flow. Mean meltabilities, over all ages, of both Cheddar and Mozzarella cheeses decreased significantly as holding time increased. Meltability of young cheese was scarcely affected by the holding time, in sharp contrast to that of the old cheese where increasing the holding time greatly reduced meltability.

Cheese↗

Reversibility of the temperature-dependent opacity of nonfat mozzarella cheese.

Salted and unsalted nonfat mozzarella cheese was made by direct acidification and stored at 4 degrees C over 60 d. Changes in cheese opacity were measured by using reflectance L* values while the cheese was heated from 10 to 90 degrees C, then cooled to 10 degrees C, and reheated to 90 degrees C. A characteristic opacity transition temperature (T(OP)) was obtained for each cheese. Both salt content and storage time influenced T(OP). Opacity during heating, cooling, and reheating formed a hysteresis. At d 1, the unsalted cheese became opaque when heated to 20 degrees C, but the salted cheese required heating to 40 degrees C. As the salted cheese was aged, its T(OP) increased so that by 60 d the cheese did not become opaque until it was heated to 70 degrees C.

Cheese↗

Invited review: perspectives on the basis of the rheology and texture properties of cheese.

Physical and chemical properties of cheese, such as texture, color, melt, and stretch, are primarily determined by the interaction of casein (CN) molecules. This review will discuss CN chemistry, how it is influenced by the cheese-making process, and how it impinges on the final product, cheese. We attempt to demonstrate that the application of principles governing the molecular interactions of CN can be useful in understanding the many physical and chemical properties of cheese and, in turn, how this can be used by the cheesemaker to produce the desired cheese. The physical properties of cheese (as well as flavor) are influenced by a number of factors including: milk composition; milk quality; temperature; the rate and extent of acidification by the starter bacteria; the pH history of cheese; the concentration of Ca salts (proportions of soluble and insoluble forms); extent and type of proteolysis, and other ripening reactions. Our hypothesis is that these factors also control and modify the nature and strength of CN interactions. The approach behind the recently proposed dual-binding model for the structure and stability of CN micelles is used as a framework to understand the physical and chemical properties of cheese.

Animals↗

Effect of calcium on microstructure and meltability of part skim mozzarella cheese.

The role of calcium in the microstructure of part skim Mozzarella cheese was evaluated using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Part skim Mozzarella cheeses with 4 calcium levels (control 0.65%, T1 0.48%, T2, 0.42%, and T3 0.35%) were manufactured and stored at 4 degrees C. Microstructure and meltability of cheeses were studied on d 1 and 30. The micrographs were analyzed for numbers, area, perimeter, roundness, and size of the fat particles. Reduced calcium cheeses had greater meltability and more hydrated protein matrix with greater number of fat particles (control=125, T1=193, T2=184, and T3=215 with SEM and control = 86, T1=87, T2= 125, and T3= 140 with CLSM). Further, area and perimeter of these fat particles were also greater in reduced calcium cheeses. Area, perimeter, and size of fat particles increased and their roundness decreased upon storage of 30 d. Decrease in free serum in the protein matrix of all cheeses upon refrigerated storage was evident from the CLSM. Hydrated protein network and better emulsified fat in low calcium cheeses might have improved melt properties of Mozzarella cheese.

Calcium↗