Impact of whey pH at drainage on the physicochemical, sensory, and functional properties of mozzarella cheese made from buffalo milk

In this study, the effects of whey pH at drainage on the physicochemical, sensory, and functional properties of mozzarella cheese made from buffalo milk during storage were investigated. Four cheese samples were manufactured using starter culture at different whey pH values [(A) 6.2, (B) 5.9, (C) 5.6, and (D) 5.3] and analyzed on the 1st, 28th, and 56th day. Ash, calcium, and phosphorus concentrations decreased as the whey pH at drainage was lowered. Cheese yield and calcium recovery were the highest in D cheeses. During storage, expressible serum levels decreased and nonexpressible serum levels increased, indicating an increase in the water holding capacity of the cheeses. Reducing the calcium content of cheeses increased meltability values, but an overly low calcium level (D cheeses) had an adverse effect on the meltability. The melting properties of cheese samples, except D cheeses, were improved with aging. A cheeses were the hardest and D cheeses the softest throughout storage. The 1st day sensory evaluations revealed that C and D cheeses were preferred and that A cheeses were not. All sensory properties of A cheeses were improved with storage. D cheeses were rated inferior to the others at the end of the storage time.     

Ethyl ester formation is enhanced by ethanol addition in mini Swiss cheese with and without added propionibacteria

Esters are important contributors to cheese flavor, but their mechanisms of synthesis in cheese are largely unknown. This study aimed to determine whether ethanol concentration limits the formation of ethyl esters in cheese. Mini Swiss cheeses were manufactured with (E) or without (C) the addition of ethanol to cheese milk. Ethanol concentrations (enzymatic analysis) were 64 +/- 17 and 330 +/- 82 microg g(-1), respectively, in C and E cheeses. E cheeses also contained 5.4 +/- 2.3 times more of the five ethyl esters quantified than C cheeses, regardless of the concentrations of esters in C cheeses (range 1-128 ng g(-1)). Furthermore, the presence of propionibacteria added as acid-producing secondary starters was associated with greater concentrations of esters, due to the increase in acid concentrations that propionibacteria induced and/or to an involvement of propionibacteria enzymes in ester synthesis. This study demonstrates that ethanol is the limiting factor of ethyl ester synthesis in Swiss cheese.     

Starter bacteria are the prime agents of lipolysis in cheddar cheese

To assess the contribution of starter lactic acid bacteria (LAB) to lipolysis in Cheddar cheese, the evolution of free fatty acids (FFAs) was monitored in Cheddar cheeses manufactured from pasteurized milks with or without starter. Starter-free cheeses were acidified by a combination of lactic acid and glucono-delta-lactone. Starter cultures were found to actively produce FFAs in the cheese vat, and mean levels of FFAs were significantly higher in starter cheeses over ripening. The contribution of nonstarter LAB toward lipolysis appears minimal, especially in starter-acidified cheeses. It is postulated that the moderate increases in FFAs in Cheddar cheese are primarily due to lack of access of esterase of LAB to suitable lipid substrate. The results of this study indicate that starter esterases are the primary contributors to lipolysis in Cheddar cheese made from good quality pasteurized milk.     

Time course and specificity of lipolysis in Swiss cheese

Controlling lipolysis in cheese is necessary to ensure the formation of desirable flavor. To get a better understanding of the mechanism of lipolysis in Swiss cheese, cheeses were manufactured with and without (control) the addition of Propionibacterium freudenreichii. Products of lipolysis were quantified throughout ripening. Half of the free fatty acids (FFA) released in milk (3.66 mg/g fat), in particular the short-chain FFA, were lost in the whey during curd drainage, whereas diglycerides and monoglycerides were retained within the curd. P. freudenreichii was responsible for the release of most FFA during ripening (10.84 and 0.39 mg/g fat in propionibacteria-containing and control cheeses, respectively). Indices of lipolysis displayed low specificity. All types of FFA were released, but butyric and palmitic acids more significantly, which could be due to a low sn-1,3 regioselectivity. All glycerides were hydrolyzed in the following order: monoglycerides>diglycerides>triglycerides. The results of this study show the quantitative and qualitative contributions of the different lipolytic agents to Swiss cheese lipolysis.     

Multiscale characterization of the organization of triglycerides and phospholipids in emmental cheese: from the microscopic to the molecular level

The chemical composition and properties of lipids, both triglycerides and phospholipids, play a major role in the functional and nutritional properties of food products. In this study, the suprastructure of fat, solid fat content, and crystallographic properties of triglycerides were investigated in hard-type cheeses from the microscopic scale to the molecular level using the combination of relevant techniques. Two industrial cheeses with different oiling off properties were compared with experimental cheeses manufactured in the laboratory. Microstructural analysis performed using confocal laser scanning microscopy showed that milk processing led to the disruption of fat globules with the formation of nonglobular fat. For a similar fatty acid composition, oiling off was mainly related to the fat in dry matter content and to the suprastructure of fat in cheese. An exogenous fluorescent phospholipid permitted the localization of milk phospholipids in the cheese matrix, which mainly remain around fat inclusions after disruption of the milk fat globule membrane, and to show heterogeneities. We also showed using differential scanning calorimetry that the suprastructure of fat did not affect the solid fat content in cheese at 4 degrees C: 71.6 +/- 4.9%. The organization of triglyceride molecules in fat crystals, elucidated at a molecular level using X-ray diffraction, corresponded to the coexistence of 2 lamellar structures (2L 40.5 angstroms and 3L 54.6 angstroms) with four polymorphic forms: alpha, two beta' and beta. A schematic representation of the multiscale organization of triglycerides and phospholipids in cheese is proposed.     

Vitamin D3 fortification, quantification, and long-term stability in Cheddar and low-fat cheeses

Considering the widespread insufficiency of vitamin D, the fortification of additional foods with vitamin D is warranted. The objective of this research was to assess the feasibility of vitamin D3 fortification in natural hard cheeses. We examined the recovery, distribution, long-term retention, and heat stability of the vitamin in industrially made fortified Cheddar and low-fat cheeses. The results indicated that the vitamin D3 did not degrade during processing, over 1 year of ripening (3-8 degrees C), or after thermal treatment at 232 degrees C for 5 min. Vitamin D3 recovery in the fortified Cheddar and low-fat cheeses were, respectively, 91 and 55% of the vitamin D3 added to the milk used to make each cheese. The remaining vitamin D3 was entrained in the whey. The vitamin D3 was uniformly distributed throughout the blocks of cheese. The fortification process did not alter the yield, chemical composition, or flavor of the Cheddar cheese. We conclude that industrially manufactured Cheddar and low-fat cheeses are suitable for vitamin D3 fortification.     

Evaluation of APHA and AOAC methods for phosphatase in cheese

Varieties of market cheese were analyzed for alkaline phosphatase by the modified rapid colorimetric method of the American Public Health Association (APHA) and the official AOAC method, 16.304-16.306. In the APHA method, 5 g cheese (pH less than 7.0) is macerated with 2 mL 1:1 carbonate buffer, or 2 mL water (for cheese with pH greater than 7.0). Addition of 0.1 mL magnesium acetate (1 mg magnesium) to test portions of cheese extracts yielded reproducible and quantitative recovery of added phosphatase. In the AOAC method, macerating 0.5 g cheese with 1 mL borate buffer before adding milk phosphatase improved recovery among cheeses. Addition of magnesium ion increased phosphatase activity in some cheeses. Phosphatases in blue mold-ripened and Swiss cheeses were inactivated by heat faster than was milk phosphatase, yet milk phosphatase added to various soft cheeses was completely inactivated at 60 degrees C for 10 min. The lability of phosphatase was due to the heat-denaturing effect of NaCl present in finished cheeses. Some Mexican style soft cheeses contained both heat-labile and heat-stable phosphatases. These data suggest that the phosphatase test to differentiate milk and microbial phosphatases on the basis of repasteurization and analysis of cheese is no longer valid.     

Identification and enumeration of beta-hemolytic Listeria monocytogenes in naturally contaminated dairy products

A DNA probe was used to identify hemolytic Listeria monocytogenes in naturally contaminated dairy products: unpasteurized milk, ricotta cheese, and imported semisoft cheeses. Of 34 milk samples, 12 were suspected to contain hemolytic L. monocytogenes; 1 contained greater than 6000 viable organisms/g. The ricotta cheese, although temperature-abused, had a titer of 3.6 x 10(6) beta-hemolytic L. monocytogenes cells/g, whereas the semisoft cheeses reached a maximum of 5.6 x 10(6) cells/g. Pure cultures of L. monocytogenes isolated from both types of cheese were found positive by the CAMP test and the DNA probe.     

Transmission electron microscopy of mozzarella cheeses made from microfluidized milk.

The nanostructure of Mozzarella cheeses prepared from microfluidized milk was compared with that of control cheeses made from untreated milk. Milk heated to 10 or 54 degrees C and containing 1.0 or 3.2% fat was homogenized by microfluidization at 34 or 172 MPa prior to cheesemaking. The effects on the casein particles and fat globules in the cheese were determined by transmission electron microscopy after 1 day and 6 weeks of storage at 4 degrees C. The micrographs showed that electron-dense regions theorized to be casein submicelles rearranged from a homogeneous configuration to a pattern of clusters during the storage period. The nanostructure of the cheeses made from milk processed under the mildest conditions resembled the controls, but otherwise the fat droplets decreased in size and increased in number as the pressure and temperature were increased. The results indicate that both homogenization temperature and pressure affect the nanostructure of Mozzarella cheese.     

Rheological and proteolytic properties of Monterey Jack goat's milk cheese during aging

To enhance the understanding of the quality traits of goat's milk cheeses, rheological and proteolytic properties of Monterey Jack goat's milk cheese were evaluated during 26 weeks of 4 degrees C storage. As expected with aging, beta-casein levels decreased with concomitant increases in peptide levels and were correlated with changes in rheological properties of the cheese. Hydrolysis of the protein matrix resulted in more flexible (increased viscoelastic properties) and softer (decreased hardness, shear stress, and shear rigidity) cheeses. During the first 4-8 weeks of storage, cheese texture changed significantly (P < 0.05) and then stabilized. Characterization of rheological and proteolytic properties of the goat's milk semihard cheese during aging provided insight into the changes occurring in the protein matrix, the relationship to structure, and a shift in cheese quality.     

Effect of different membrane separation technologies (Ultrafiltration and microfiltration) on the texture and microstructure of semihard low-fat cheeses

Semihard low-fat cheeses made from ultrafiltered (UF) or microfiltered (MF) milk were compared. The use of MF membranes and milder pasteurization of the milk reduced the retention of whey proteins in the retentate to 35%, compared with approximately 100% retained in the UF process. Microbiological development, physicochemical composition, and cheese ripening were not altered by the concentration processes. The lower retention of whey protein in MF cheeses accounted for their higher hardness, which correlated with higher firmness values in the textural analysis. Microstructure showed a protein matrix with open spaces through the protein network, although micrographs of UF cheeses showed the presence of spongy structures linked to the casein, which did not appear in MF cheeses and which correspond to the denatured whey protein bound to the casein. Firmness was scored better in MF cheeses, although when MF membranes were used, the optimum yields achieved using UF membranes were not attained.     

Flavor enhancement of reduced fat cheddar cheese using an integrated culturing system

Mild cheese flavor in reduced fat Cheddar cheese was enhanced by using an integrated starter culture system. Three cultures, Lactococcus lactis subsp. cremoris SK11, L. lactis subsp. lactis biovar. diacetylactis JVI, and Lactobacillus casei 7A, were carefully selected to obtain a nonbitter, mildly acid, buttery flavored cheese. Cheeses were produced from all possible combinations of these cultures with the constraint that L. lactis subsp. cremoris SK11 was used as the primary acid-producing culture. Cheeses made with SK11 were compared to cheeses produced using an L. lactis subsp. cremoris commercial starter culture. Cheeses were ripened for 150 days and periodically sampled for chemical, microbiological, and sensory analysis. Cheeses produced with L. lactis subsp. cremorisSK11 had substantially lower bitterness intensity than the cheeses produced with commercial starter culture. L. lactis subsp. lactis biovar. diacetylactis JVI significantly increased diacetylacetoin and acetate concentrations. Sensory results indicate that these cheeses had increased buttery (diacetyl) flavor.     

Use of recombinant cyprosin in the manufacture of ewe's milk cheese

A recombinant cyprosin from the cardoon (Cynara cardunculus L.) was assayed and compared with calf rennet in batches of ewes' milk cheese by determining different chemical, biochemical, and microbiological parameters over 4 months of ripening. There were no differences between the two types of coagulants in most chemical parameters, a(w), and pH. Proteolysis was more marked and rapid in cheese containing recombinant cyprosin as coagulant, the soluble nitrogen content of which was significantly higher (p < 0.01) than that of the cheese obtained with animal rennet; at the end of ripening the nonprotein nitrogen of cheese produced with recombinant cyprosin was slightly higher (p > 0.05) as compared with that in cheeses obtained with animal rennet. Microbial counts in the milk used for making cheese were high in most of the groups analyzed. Despite slight differences in counts, the main microbial groups analyzed were similar in cheese produced with both types of coagulants throughout ripening.     

Chemical and microbiological characteristics of ewes' milk cheese manufactured with extracts from flowers of Cynara cardunculus and Cynara humilis as coagulants

The chemical and microbial characteristics as well as the flavor and aroma of Los Pedroches cheese made using aqueous extracts of Cynara cardunculus L. flowers were compared with those of cheeses manufactured with extracts of Cynara humilis L. throughout ripening. The two thistle species assayed were found to have no appreciable effect on the moisture, fat, protein, and NaCl contents of the cheese or on its water activity, flavor, and aroma; however, the use of C. humilis resulted in reduced lactic acid content (p < 0.001) and higher pH values (p < 0.05) relative to those of cheese specimens produced with C. cardunculus. The protein breakdown of the cheeses was assessed in terms of soluble nitrogen (SN), nonprotein nitrogen (NPN), and amino acid nitrogen (AAN). Proteolysis was more marked and rapid in cheese containing C. cardunculus as coagulant, the SN and NPN contents of which were significantly higher (p < 0. 01) than those of the cheese obtained with the species C. humilis; AAN contents were similar in both species of Cynara throughout ripening. Although total viable, coliform, and lactobacilli counts were similar in cheeses produced with both types of plant coagulant throughout ripening, enterobacteria and yeasts counts (p < 0.01) and molds counts (p < 0.05) were higher in cheese produced with C. humilis than in cheese obtained with C. cardunculus.     

Milk excretion of ivermectin and moxidectin in dairy sheep: assessment of drug residues during cheese elaboration and ripening period

Ivermectin (IVM) and moxidectin (MXD) are broad-spectrum endectocide antiparasitic drugs extensively used in food-producing animals. The patterns of IVM and MXD excretion in milk were comparatively characterized following their subcutaneous administration (200 microg.kg(-1) of body weight) to lactating dairy sheep. The relationship between milk excretion and plasma disposition kinetics of both compounds was characterized. A pool of milk collected from all of the animals in each experimental group was used for cheese elaboration. IVM and MXD residual concentrations were assessed during the cheese-making process and ripening period. IVM and MXD concentrations were measured in plasma, milk, and milk product (whey, curd, and cheese) samples using an HPLC-based methodology with fluorescence detection. IVM and MXD were extensively distributed from the bloodstream to the mammary gland, and large quantities, particularly of MXD, were excreted in milk. Residual concentrations of both compounds were recovered in milk up to 30 (IVM) and 35 (MXD) days post-treatment. The total fraction of the administered dose excreted in milk for MXD was significantly higher than that of IVM. During cheese production, the highest residual concentrations of both molecules were measured in the curd. Thirty-four percent of the total drug residue measured in the pooled milk collected from treated sheep was lost during the cheese-making process. The lowest residual concentrations were measured in the whey. IVM and MXD concentrations in the elaborated cheese tended to increase during the ripening period, reaching the highest residual level at 40 days of cheese maturation. The long persistence of milk residual concentrations of MXD and IVM in lactating dairy sheep and the high concentrations found in cheese and other milk-related products should be seriously considered before recommendation of the extralabel use of these antiparasitic drugs in dairy animals.     

Volatile compounds in Hispánico cheese manufactured using a mesophilic starter,a thermophilic starter,and bacteriocin-producing Lactococcus lactis subsp. lactis INIA 415

The effect of the addition of Lactococcus lactis subsp. lactis INIA 415, a strain harboring the structural genes of nisin Z and lacticin 481, on the formation of volatile compounds in Hispánico cheese manufactured with a mesophilic starter or with the mesophilic starter and a thermophilic starter was investigated. Addition of bacteriocin-producing L. lactis subsp. lactis INIA 415 to milk enhanced the formation of 2-methyl-propanal, 2-methylbutanal, 3-methylbutanal, 2-methyl-1-propanol, 3-methyl-1-butanol, 1-octanol, 2-butanone, and 2,3-butanedione. On the other hand, addition of thermophilic starter enhanced the formation of acetaldehyde, ethanol, 3-methyl-2-buten-1-ol, ethyl butanoate, ethyl hexanoate, 2-butanone, and 2,3-butanedione in Hispánico cheese. Stepwise discriminant analysis using the relative abundances of volatile compounds classified cheeses by type of starter, with function 1 related to thermophilic starter and function 2 to bacteriocin producer.     

Characterization of aroma compounds responsible for the rosy/floral flavor in Cheddar cheese

The aroma-active compounds that contribute to the rosy/floral flavor in Cheddar cheese were characterized using both instrumental and sensory techniques. Two cheeses (>12 months old) with rosy/floral flavor and two Cheddar cheeses of similar ages without rosy/floral flavors were selected. After direct solvent extraction/solvent-assisted flavor evaporation and separation into neutral/basic and acidic fractions, samples were analyzed by gas chromatography-olfactometry with aroma extract dilution analysis. Selected compounds were quantified using internal standard methodology. Some of the intense aroma-active compounds in the neutral basic fraction of the rosy/floral cheeses included 2-phenethanol (rosy), phenylethyl acetate (rosy), and phenylacetaldehyde (rosy/floral). Quantification, threshold analysis, and sensory analysis of model cheeses confirmed that increased concentrations of phenylacetaldehyde and phenylacetic acid caused rosy/floral flavor when spiked into Cheddar cheese.     

Enzyme immunoassay for mycophenolic acid in milk and cheese

Mycophenolic acid (MPA) was reacted with N-hydroxysuccinimide and conjugated to keyhole limpet hemocyanin (KLH), and to horseradish peroxidase (HRP), respectively. The MPA-KLH was used to produce anti-MPA antiserum in rabbits. A competitive direct enzyme immunoassay (EIA) for MPA was established with anti-MPA antiserum and MPA-HRP conjugate. The mean 50% inhibition and detection limit of MPA standard curves (n = 103) were 197 +/- 67 and 81 +/- 48 pg/mL, respectively. The EIA was specific for MPA and its synthetic 2-morpholinoethyl ester, mycophenolate mofetil (91% relative cross-reactivity). Raw bulk milk and pasteurized milk, with and without beta-glucuronidase pretreatment, were analyzed by EIA. No MPA was found in milk, at a detection limit of 100 pg/mL (recovery 58-66% at 0.125-2 ng/mL). Blue-veined cheese from the German market (n = 53) was analyzed by EIA, and the detection limit was at 0.5 ng/g (recovery 68-79% at 5-100 ng/g). All but two cheeses contained MPA, although mostly (66%) at levels of <10 ng/g. MPA at 400-1200 ng/g was found in Roquefort cheeses. Highest levels (4-11 microg/g) were found in a German soft cheese preparation. MPA levels in mycelium-rich parts of cheese were 3 times higher than in mycelium-free parts.     

Volatile compounds produced in cheese by pseudomonas strains of dairy origin belonging to six different species

Production of volatile compounds by seven Pseudomonas strains belonging to six different species, Ps. brenneri, Ps. graminis, Ps. libanensis, Ps. lundensis, Ps. putida, and Ps. rhodesiae, was investigated, with the aim of elucidating their possible contribution to the volatile profile of cheese. Laboratory-scale cheeses were made from pasteurized milk of low bacterial counts separately inoculated with approximately 10(5) colony-forming units/mL of each Pseudomonas strain and ripened for 12 days at 10 degrees C. A total of 122 volatile compounds were identified in cheeses by GC-MS of the dynamic headspace. The abundance of 62 compounds, belonging to eight chemical groups (aldehydes, ketones, acids, esters, alcohols, hydrocarbons, benzene compounds, and sulfur compounds) increased during ripening for at least one of the strains. Most groups of volatile compounds were more abundant in the outer part of cheeses than in the inner part, in agreement with the aerobic metabolism of the genus Pseudomonas and coinciding with the higher counts in the outer part. Production of volatile compounds in cheese by Pseudomonas was shown to be species-dependent.     

Typicality and geographical origin markers of protected origin cheese from The Netherlands revealed by PTR-MS

Volatile fingerprints of 30 cumin cheese samples of artisanal farmers' cheese of Leiden with EU Protected Designation of Origin (PDO) and 29 cumin cheese samples of varying commercial Dutch brands without PDO protection were used to develop authentication models. The headspace concentrations of the volatiles, as measured with high sensitivity proton-transfer mass spectrometry, were subsequently subjected to partial least-squares discriminant analysis (PLS-DA). Farmers' cheese of Leiden showed a distinct volatile profile with 27 and 9 out of the 60 predominant ions showing respectively significantly higher and lower concentrations in the headspace of the cheese in comparison to the other cumin cheeses. The PLS-DA prediction models developed classified in cross-validation 96% of the samples of PDO protected, artisanal farmers' cheese of Leiden correctly, against 100% of commercial cumin cheese samples. The characteristic volatile compounds were tentatively identified by PTR-time-of-flight-MS. A consumer test indicated differences in appreciation, overall flavor intensity, creaminess, and firmness between the two cheese groups. The consumers' appreciation of the cumin cheese tested was not influenced by the presence of a name label or PDO trademark.