Use of high-pressure-treated milk for the production of reduced-fat cheese

Pasteurized (65°C, 30 min), pressurized (400 MPa, 22°C, 15 min) and pasteurized–pressurized milks were used for reduced-fat (approximately 32% of total solids) cheese production. Pressurization of milk increased the yield of reduced-fat cheese through an enhanced β-lactoglobulin and moisture retention. In addition, pressurisation of pasteurized skim milk improved its coagulation properties. The cheeses made from pasteurized–pressurized and pressurized milks showed a faster rate of protein breakdown than the cheese made from pasteurized milk, that might be mainly attributed to a higher level of residual rennet. Hardness of the experimental cheeses, as determined by both the sensory panel and instrumental analyses, decreased as the moisture content and proteolytic degradation of the cheese increased (pasteurized>pressurized>pasteurized–pressurized). In general terms, pressurization of reduced-fat milk prior to cheese-making improved cheese texture and thus accounted for a higher overall acceptability, except for the cheeses made from pasteurized–pressurized milk at 60 d of ripening, whose acceptability score was adversely affected by bitterness.     

Lamb rennet paste in ovine cheese manufacture. Lipolysis and flavour

In a preliminary study with commercial ewe's milk cheeses, there were statistically significant differences in the sensory evaluation scores and the amounts of short-chain (C4–C10) free fatty acids (FFAs) between cheeses made with lamb rennet paste or bovine rennet. Experimental ewe's milk cheeses were manufactured with 2 levels of artisanally produced lamb rennet paste and 2 levels of bovine rennet in 50 L vats, with a manufacturing replicate done within 1 week. Total coagulating activity was 2500 RU for cheeses manufactured with a high amount of rennet or 1000 RU for cheeses manufactured with a low amount of rennet. The two batches of lamb rennet pastes used had 4.0 (early Spring) and 6.5 U g⁻¹ lipase (late Spring), whereas no lipase activity was detected in bovine rennets. The total concentration of FFAs in cheeses manufactured with lamb rennet paste was significantly higher than in cheeses manufactured with bovine rennet, regardless of ripening time and time of the year. Lipolysis in the former cheeses increased with the total units of lipolytic activity added. The increase in lipolysis in cheeses made with lamb rennet was primarily due to higher amounts of short-chain fatty acids (C4–C10). Butyric acid was the main FFA in cheeses made with lamb rennet paste, representing 34.5 μmol 100 μmol⁻¹ (low level of rennet) and 44.2 μmol 100 μmol⁻¹ (high level of rennet) of the total FFAs, whereas it represented only 17 μmol 100 μmol⁻¹ of the total FFAs in all cheeses made with bovine rennet. The concentration of individual FFAs of chain length <C12 were significantly higher in cheeses made with lamb rennet paste than in cheeses made with bovine rennet. Cheeses made with lamb rennet paste received significantly higher intensity scores for odour and flavour intensity, sharp odour, ‘piquant’ and bitter flavours and ‘natural rennet’ odour and flavour.     

High-pressure-induced changes in the rennet coagulation properties of bovine milk

The mechanism of high-pressure (HP)-induced changes in rennet coagulation properties of milk, particularly the role of whey protein-casein micelle associations, was studied. Treatment at 100 or 250 MPa reduced the rennet coagulation time (RCT) of raw skimmed bovine milk, compared with untreated milk. Treatment at 400 MPa had little effect, but at 600 MPa, RCT increased considerably. HP-induced increases in RCT did not occur in serum protein-free milk or milk treated with the sulphydryl-oxidising agent KIO₃, which prevents association of denatured β-lactoglobulin with casein micelles. Treatment at 5 or 10 °C at 250–600 MPa resulted in shorter RCT than treatment at 20 °C. In milk without KIO₃, coagulum strength was highest after treatment at 250 or 400 MPa, whereas in milk with KIO₃ it was highest after treatment at 400 MPa. These results indicate the significance of HP-induced association of whey proteins with casein micelles for rennet coagulation properties of milk.     

Effect of On-Farm Heating and Storage of Milk on Cottage Cheese Yield

Effect of on-farm heat treatment of milk on cottage cheese yields was studied. Fresh, raw milk heated to 74°C for 10 s was cooled and stored for 7 days at 3°C. Control and experimental lots of milk were separated and pasteurized at 72°C for 15 s and were used to make cottage cheese. Microbiological, shelf-life, flavor, and texture studies showed the experimental lots of cheese were as good as or better than control lots. Yield of cottage cheese was significantly higher when made from heated milk.     

Bovine milk procathepsin D: Presence and activity in heated milk and in extracts of rennet-free UF-Feta cheese

Milk samples were heat-treated at 72, 85 and 99°C for 15 or 60 s, and the effect on the stability of the milk acid proteinase zymogen procathepsin D was studied by combining immunoblotting using antibodies directed against bovine cathepsin D and its propeptide and by measuring residual procathepsin D-derived activity. Approximately half of the procathepsin D-derived activity detected in milk serum remained after heat treatment at 72°C/15 s or 72°C/60 s, while heat treatment at increased temperature further reduced the detectable activity. In accordance, immunoreactive procathepsin D was detected in serum from milk heated at 72°C/15 s and 72°C/60 s, while very low amounts of immunoreactivity were observed after treatment at higher temperatures. Contrary to the decrease in milk serum, the amount of procathepsin D antigen associated with casein micelles slightly increased with the temperature of the heat treatment, but still the measurable proteolytic activity derived from procathepsin D in the casein micelle samples decreased with temperature treatment. Moreover, the presence of procathepsin D and derived proteolytic activity was demonstrated in rennet free UF-Feta cheese. These results correlated with the finding of α_s1-I and para-κ-caseins in rennet free cheese. This is the first demonstration of procathepsin D in cheese, and of activity derived from indigenous procathepsin D in milk contributing to the proteolysis process in UF-products.     

Rennet-induced coagulation of enzymatically cross-linked casein micelles

The enzymatic cross-linking of casein micelles with transglutaminase had an adverse influence on rennet-induced coagulation. Incubation with transglutaminase at 30 °C progressively reduced the levels of monomeric caseins and increased rennet flocculation time (RFT) in a Berridge test. For incubation up to 3 h at 30 °C, the reciprocal of the RFT was linearly correlated with the level of residual monomeric κ-casein, indicating that at complete cross-linking flocculation is absent. After treatment for 4–24 h at 30 °C, no residual monomeric κ-casein was detected and no rennet-induced flocculation of the casein micelle suspension was observed. Monitoring rennet-induced coagulation by diffusing wave spectroscopy revealed that transglutaminase-induced inhibition of rennet-induced coagulation of casein micelles is primarily due to an inhibition of the secondary phase of rennet coagulation, i.e., the gelation and gel-firming phase of the casein micelle coagulation. The gelation and fusion of κ-casein-depleted para-casein micelles as in normal milk appears to be absent if the casein macropeptide remains attached to the casein micelle.     

Evolution of lipolysis during the ripening of traditional Feta cheese

Lipolysis was studied during ripening of traditional Feta cheese produced in two small dairies, A and B. The cheeses were made from a thermized mixture of ewes’/goats’ milk by using yoghurt as starter and artisanal rennet from lambs’ and kids’ abomasa (cheese A) or mixed artisanal rennet with calf rennet (cheese B).The acid degree value and the free fatty acids (FFA) contents in both cheeses increased sharply up to 18 d (pre-ripening period at 15 °C) and continued to increase throughout ripening. In both mature cheeses, acetic acid was found at high levels (13–18% of the total FFAs). However, except for this, all FFA contents differed significantly (P < 0.05) between the two cheeses throughout ripening. The levels of individual and total C2:0–C8:0, C10:0–C14:0 and C16:0–C18:2 fatty acids were significantly higher (P < 0.05) in cheese A than in cheese B. Presumably the difference, especially in the C2:0–C8:0 content, was due mainly to the type of the rennet used. Butyric acid was the dominant FFA in cheese A (20% of the total FFAs at 120 d), while the most abundant FFAs in cheese B were capric (18%) and lauric acid (18%). In general, the lipolysis degree of the two cheeses was higher than those reported for the industrially-made Feta cheese.In organoleptic evaluation, cheese A had a piquant taste that was attributed to its high content of butyric acid and showed a significantly (P < 0.05) higher total score than cheese B.     

Acid coagulation properties and suitability for yogurt production of cows’ milk treated by high-pressure homogenisation

The effects of high-pressure homogenisation (HPH) of cows’ milk were investigated for suitability for yogurt manufacture, compared with the processes currently applied in industry. Milk at different inlet temperatures (30 °C or 40 °C) was subjected to HPH treatment at 100, 200 or 300 MPa (one stage) and 130, 230 or 330 MPa (two-stage). HPH-treated milk was compared with milk heat-treated (90 °C for 90 s) and homogenised at 15 MPa, and with milk treated under the same thermal conditions and also fortified with 3% skim milk powder. Milk treated at 300 or 200 MPa showed higher gel strengths on coagulation, higher gel firmness in texture analysis, less syneresis and lower titratable acidity compared with conventionally treated milk fortified with 3% skim milk powder.     

Partial renneting of pasteurised bovine milk: Casein micelle size,heat and storage stability

The effects of partial renneting at low temperature on the casein micelle (CM) size and the storage stability of milk were investigated. Low chymosin concentrations (≤ 0.03 IMCU mL^− 1) was applied to pasteurised skim milk at 4 °C and enzyme activity was terminated by thermal application at 60 °C/3 min and 85 °C/30 min, referred to as low heat (LHT) and high heat (HHT) treatment milk, respectively. The addition of rennet with concentrations of 0.01, 0.02 and 0.03 IMCU mL^− 1 for 15 min resulted in κ-casein hydrolysis of 10, 20 and 25%, respectively. Moreover, mean CM size of milk was reduced by up to 10 nm. For LHT milk, the renneted micelles appeared to be stable for up to 17 days, especially in response to the application of 0.01 IMCU mL^− 1 and at a storage temperature of 4 °C. Severe heating at 85 °C/30 min to inactivate the enzyme caused an increase in CM size.     

The influence of lactoperoxidase,heat and low pH on survival of acid-adapted and non-adapted Escherichia coli O157:H7 in goat milk

In hot climates where quality of milk is difficult to control, a lactoperoxidase (LP) system can be applied in combination with conventional preservation treatments at sub-lethal levels to inhibit pathogenic microbes. This study investigated the effect of combined heat treatments (55 °C, 60 °C and 72 °C) and milk acidification (pH 5.0) on survival of acid-adapted and non-adapted Escherichia coli O157:H7 strains UP10 and 1062 in activated LP goat milk. Heat treatment at 72 °C eliminated E. coli O157:H7. Acid-adapted strains UP10 and 1062 cells showed resistance to combined LP and heat at 60 °C in fresh milk. The inhibition of acid-adapted and non-adapted E. coli O157:H7 in milk following combined LP-activation, heat (60 °C) and milk acidification (pH 5.0) suggests that these treatments can be applied to reduce E. coli O157:H7 cells in milk when they occur at low numbers (<5 log₁₀ cfu mL^?1) but does not eliminate E. coli O157:H7 to produce a safe product.     

An investigation of the plasmin–plasminogen system in caprine milk and cheese

The variability in activities of plasmin (PL) and plasminogen (PG) in bulk milk samples from different breeds of goats was investigated. The mean PL activity was higher (19.56±4.72 U g⁻¹) than in milk from other ruminant species, while PG-derived activity was, surprisingly, lower (12.84±5.31 U g⁻¹). No significant differences in PL and PG levels were observed among goat breeds, but PL activity and PL/PG ratio increased in late lactation compared with early lactation. PL activities in pilot-scale curds (acid or rennet) and in some Italian cheeses made from caprine milk were also measured. Generally, acid curds and cheeses had lower residual PL and PG-derived activities than rennet curds and semi-hard cheeses. In addition, the PL/PG ratio was greatly reduced in rennet curds, due to a higher extent of PG-derived activity. Measurement of proteolysis of β-casein in curds confirmed that more extensive proteolysis occurred in rennet curds, which had higher residual PL activities.     

Evaluation of pulsed electric field and minimal heat treatments for inactivation of pseudomonads and enhancement of milk shelf-life

Pulsed Electric Field (PEF) treatment of milk provides the opportunity to increase the shelf-life of fresh milk for distribution to distant markets. PEF treatments were evaluated in sterile (UHT) milk to determine the inactivation of added spoilage Pseudomonas isolates and the subsequent gains in microbial shelf-life (time taken to reach 10⁷ CFU mL^− 1). Little inactivation of Pseudomonas was achieved at 15 or 40 °C compared with 50 or 55 °C. The greatest inactivation (> 5 logs) was achieved by processing at 55 °C with 31 kV cm^− 1 (139.4 kJ L^− 1). Heat treatment at the application temperature without PEF treatment caused minimal inactivation of Pseudomonas (only 0.2 logs), demonstrating that the inactivation of the Pseudomonas was due to the PEF treatment rather than the heat applied to the milk. At added Pseudomonas levels of 10³ and 10⁵ CFU mL^− 1, the microbial shelf-life of PEF-treated milk was extended by at least 8 days at 4 °C compared with untreated milk. The total microbial shelf-life of the PEF-treated milk was 13 and 11 days for inoculation levels of 10³ and 10⁵ CFU mL^− 1 respectively. The results indicate that PEF treatment is useful for the reduction of pseudomonads, the major spoilage bacteria of milk.Industrial relevancePseudomonads are the major psychrotrophic spoilage microflora of refrigerated, stored HTST pasteurised milk. Long-life (UHT) products are an important component of milk sales in South-East Asia, but in recent years there has been an increasing demand for less processed milk products with extended shelf-life. The recent practice of shipping fresh bulk milk from Australia to South-East Asian countries has necessitated additional heat treatment prior to export and on arrival, to achieve the required shelf-life. Pulsed electric field treatment of HTST milk, applied alone or in combination with mild heat under optimised conditions, offers the opportunity of shelf-life extension, while limiting the reduction in quality attributes of milk associated with more severe additional heat treatments.     

Proteolysis in Milk Associated with Increasing Somatic Cell Counts

Individual cow samples were collected and preserved with potassium dichromate. Somatic cells counts were determined. Tyrosine value was used as an index of proteolysis. Sixty-six samples ranged in somatic cell count from < 50,000 to > 2,000,000/ml. Initial milk tyrosine values and tyrosine values for milks incubated for 24 h at 37°C showed proteolytic activity increased with increasing somatic cell count. The increase in proteolysis in preserved milk refrigerated for 72 h at 6.7°C was over 1.5 times greater in milks with > 1,000,000 cells/ml than in milks with < 60,000 cells/ml. When preserved milks were laboratory pasteurized, cooled, and stored at 6.7°C for 14 d, some proteolytic activity was detected in milks at all concentrations of somatic cells, and proteolysis increased as somatic cell counts increased. Laboratory-pasteurized samples of milk with various somatic cell counts were also incubated at 30°C for 3 and 6 h to duplicate the proteolysis that could occur during the ripening, coagulation, cutting, and cooking steps of cheese making. Again, the greatest increases in tyrosine were in milks with high somatic cell count. Protease(s) associated with elevated somatic cell counts will damage raw milk quality upon storage, pasteurized fluid milk over shelf-life, and milk during cheese making.     

Influence of lupin-based milk alternative heat treatment and exopolysaccharide-producing lactic acid bacteria on the physical characteristics of lupin-based yogurt alternatives

In the present study we investigated the influence of heat treatment of lupin-based (LB) milk alternatives and different exopolysaccharide (EPS)-producing lactic acid bacteria on the physical characteristics of set-type LB yogurt alternatives. LB milk alternatives, obtained from protein isolate of Lupinus angustifolius cv. Boregine, were either pasteurized at 80 °C for 60 s or ultra-high temperature (UHT) heated at 140 °C for 10 s and was fermented with Lactobacillus plantarum TMW 1.460 and 1.1468, Pediococcus pentosaceus BGT B34 and Lactobacillus brevis BGT L150. Fermentation duration was strongly affected by heat treatment: different strains needed between 25 to 35 h in UHT LB milk alternative to reach a pH of 4.5 compared to 14 to 24 h in pasteurized LB milk alternative. EPS extraction revealed slightly higher amounts of EPS for UHT LB yogurt alternatives (~ 0.5–0.9 g/l; pasteurized: ~ 0.4–0.7 g/l). The more intensive heat treatment (UHT) resulted also in better rheological (apparent viscosity, hysteresis loop area, flow point, elastic, viscous and complex modulus) and textural properties (firmness, consistency, cohesiveness and index of viscosity) of the investigated LB yogurt alternatives. Furthermore, LB yogurt alternatives out of UHT milk alternative revealed a lower tendency to syneresis, measured with siphon and centrifugation method. This work contributes to the fundamental knowledge of the textural properties of LB yogurt alternatives.     

Gelation Profiles of Yogurt as Affected by Heat Treatment of Milk

Milk was heated at 140°C for 6 s (UHT), 98°C for 1.87 min (HTST) and 85°C for 10 min (vat-pasteurized), homogenized, inoculated with yogurt culture, and incubated at 45°C. During incubation, pH, ionic Ca, and viscosity changes were continuously monitored by interfacing to a microcomputer. Samples for transmission electron microscopy were taken at selected pH intervals.Acid fermentation of milk was a multistage process comprising 1) an initial lag period of low viscosity; 2) a period of rapid viscosity change, and 3) a stage of high constant viscosity. The shapes of the viscosity versus incubation time curves were the same for all heat treatments differing only in the extent of viscosity change in stage 2. A microstructural study of UHT milk indicated that at pH 5.1, casein micelles appeared to dissociate into subparticles of approximately 30 to 40 nm diameter. By pH 4.8, subparticles reassociated into large casein aggregates of nonspecific shape and dimensions. Micellar dissociation was thought to be influenced by conversion of colloidal Ca to ionic Ca.     

Manufacture of Cheddar cheese from high-pressure-treated whole milk

The cheese-making characteristics of high-pressure (HP)-treated milk were examined. The rennet coagulation time of pasteurised milk decreased after HP treatment at 400 MPa but increased after treatment at 600 MPa. The L-value (whiteness) of milk decreased directly after HP treatment but, over the course of coagulation, whiteness of HP-treated milk increased to the same level as in the control. Cheddar cheese was then manufactured from raw whole milk or whole milk treated by high-pressure (HP) at 400 MPa (HP400) or 600 MPa (HP600) for 10 min at 20 °C. HP treatment of raw milk at 600 MPa resulted in a 3.66 log reduction in the initial counts of non-starter lactic acid bacteria (NSLAB), decreased protein and fat content, as well as a lower pH compared to the control. Furthermore, higher treatment pressures resulted in increased incorporation of β-lactoglobulin into the cheese curd, with parallel increases in yield by 1.23% and 7.78% for HP400 and HP600 cheeses, respectively. Overall, this study showed that the effects of HP treatment on milk proteins increased rennet coagulation times and changes in cheese composition at day 1.Industrial relevanceHigh-pressure treatment is a novel technology which has been applied to a number of commercial food products. In this study, HP-induced changes in milk proteins resulted in increased cheese yields and increased cheese whiteness. In addition, HP treatment significantly reduced the microflora of raw milk cheese. Those attributes could be of interest for both industry and consumer.     

Properties of low-fat stirred yoghurts made from high-pressure-processed skim milk

Physical properties of stirred yoghurt made from reconstituted skim milk that was high-pressure (HP)-treated at 100, 250 or 400 MPa, at 25, 70 or 90 °C, for 10 min, prior to inoculation with yoghurt cultures, were studied; portions of milk HP-treated at 25 °C were also heat-treated at 90 °C for 10 min before or after pressure treatment. Control yoghurts were made from skim milk given a heat treatment at 90 °C for 10 min. Fermentation time was not affected by treatment applied to the milk. HP treatment of skim milk at 25 °C, before or after heat treatment, gave stirred yoghurts of similar viscosities to that made from conventionally heat-treated milk. Lower viscosities were obtained when stirred yoghurts were made with milk HP-treated at elevated temperatures. A model is proposed to correlate properties of yoghurt with HP/heat-induced changes in interactions and structures of protein in the milk samples.Industrial relevanceTo meet end user expectations, the dairy industry needs to diversify its product range by tailoring specific functionalities. To meet these expectations, new processing methods such as high-pressure processing are of interest for their potential to achieve specific and/or novel functionalities and/or improve efficiencies, including reduced chemical and water use. In this paper, an investigation of the use simultaneous pressurization and heating of milk before the manufacture of stirred yoghurt is presented.     

Survival of Listeria monocytogenes in Galotyri,a traditional Greek soft acid-curd cheese,stored aerobically at 4°C and 12°C

Galotyri is a traditional Greek soft acid-curd cheese, which is made from ewes’ or goats’ milk and is consumed fresh. Because cheese processing may allow Listeria monocytogenes post-process contamination, this study evaluated survival of the pathogen in fresh cheese during storage. Portions (0.5 kg) of two commercial types (<2% salt) of Galotyri, one artisan (pH 4.0±0.1) and the other industrial (pH 3.8±0.1), were inoculated with ca. 3 or 7 log cfu g⁻¹ of a five-strain cocktail of L. monocytogenes and stored aerobically at 4°C and 12°C. After 3 days, average declines of pathogen's populations (PALCAM agar) were 1.3–1.6 and 3.7–4.6 log cfu g⁻¹ in cheese samples for the low and high inocula, respectively. These declines were independent (P>0.05) of the cheese type or the storage temperature. From day 3, however, declines shifted to small or minimal to result in 1.4–1.8 log cfu g⁻¹ of survivors at 28 days of storage of all cheeses at 4°C, indicating a strong “tailing” independent of initial level of contamination. Low (1.2–1.7 log cfu g⁻¹) survival of L. monocytogenes also occurred in cheeses at 12°C for 14 days, which were prone to surface yeast spoilage. When ca. 3 log cfu g⁻¹ of L. monocytogenes were inoculated in laboratory scale prepared Galotyri of pH ≅4.4 and ≅3% salt, the pathogen died off at 14 and 21 days at 12°C and 4°C, respectively, in artisan type cheeses fermented with the natural starter. In contrast, the pathogen survived for 28 days in cheeses fermented with the industrial starter. These results indicate that L. monocytogenes cannot grow but may survive during retail storage of Galotyri despite its low pH of or slightly below 4.0. Although contamination of Galotyri with L. monocytogenes may be expected low (<100 cfu g⁻¹) in practice, that long-term survival of the pathogen in commercial cheeses was shown to be unaffected by the artificial contamination level (3 or 7 logs) and the storage temperature (4°C or 12°C), which should be a concern.     

A preliminary study on the role of alkaline phosphatase in cheese ripening

A preparation of exogenous alkaline phosphatase (ALP), containing 17,500 mU L⁻¹, was added to pasteurized milk (PM) to study its role in cheese ripening. Three miniature Cheddar-type cheeses were made from PM containing no added ALP (control), PM plus 23 μL ALP (T1), to give ALP concentration similar to that in raw milk, and PM plus 46 μL ALP (T2). Milk, after addition of ALP, was held at 6 °C for 12 h before cheese manufacture and the experiment was replicated three times. The control, T1 and T2 milks contained ALP activity of 415, 2391 and 4705 mU L⁻¹, respectively. The addition of ALP to PM caused significant (P<0.05) changes in moisture content of miniature cheeses but did not cause any changes in protein content. Levels of water-soluble N during ripening of the cheeses were similar for control, T1 and T2 cheeses. The concentration of amino acids was not affected by the level of ALP present in milk. However, reversed-phase HPLC showed differences in the peptide patterns of control, T1 and T2 cheeses, suggesting a role of ALP in cheese ripening. The results suggest that ALP may play a role in cheese ripening, but further studies are needed to confirm this.     

The use of sedimentation field-flow fractionation in the size characterization of bovine milk fat globules as affected by heat treatment

Size distribution of fat globules affects the appearance, taste and stability of milk and milk-based products. Full-fat, semi-fat and chocolate bovine milk were subjected to heat treatment within a temperature range of 50–125 °C for 1 h. Sedimentation field-flow fractionation was employed to determine the changes in mean particle diameter of milk fat globules as affected by heat treatment. The mean particle diameter of fat droplets increased with increasing heating temperature for most samples. The particle size of fat globules increased on average 40 nm (4.65%) for full-fat and 72 nm (8.52%) for semi-fat milk following the heat treatment (50–125 °C). Chocolate milk exhibited considerable increase in particle size (104 nm, 12.53%) within a certain temperature range (50–110 °C), followed by a decrease in particle size when heated at 125 °C for 1 h. Heat-induced flocculation due to attractive interactions between hydrophobic sites on denatured protein molecules on different droplets was assumed to be mainly responsible for the increases in particle size observed in this study. Extensive heat-induced denaturation of milk proteins was also indicated by Native PAGE. Sedimentation field-flow fractionation proved to be a useful technique for adequately monitoring heat-induced changes in particle size distributions in milk.