A review of the microbiological hazards of dairy products made from raw milk

This review concentrates on information concerning microbiological hazards possibly present in raw milk dairy products, in particular cheese, butter, cream and buttermilk. The main microbiological hazards of raw milk cheeses (especially soft and fresh cheeses) are linked to Listeria monocytogenes, verocytotoxin-producing Escherichia coli (VTEC), Staphylococcus aureus, Salmonella and Campylobacter. L. monocytogenes, VTEC and S. aureus have been identified as microbiological hazards in raw milk butter and cream albeit to a lesser extent because of a reduced growth potential compared with cheese. In endemic areas, raw milk dairy products may also be contaminated with Brucella spp., Mycobacterium bovis and the tick-borne encephalitis virus (TBEV). Potential risks due to Coxiella burnetii and Mycobacterium avium subsp. paratuberculosis (MAP) are discussed. Pasteurisation ensures inactivation of vegetative pathogenic microorganisms, which increases the safety of products made thereof compared with dairy products made from raw milk. Several control measures from farm to fork are discussed.     

Cycled high hydrostatic pressure processing of whole and skimmed milk: Effects on physicochemical properties

In this study, we compare the effects of single- and double-cycle HP treatments at 600 MPa on inactivation of the natural microflora and physicochemical properties of whole and skimmed milk of high bacterial load. The results show that two-cycled HP (2 × 2.5 min) was more effective (P < 0.05) on microbial inactivation, and caused similar or slightly less changes (P > 0.05) in physicochemical properties of milk in comparison to single cycled HP (1 × 5 min). In addition to the expected milk protein structure changes, HP at 600 MPa caused only slight effects on milk fat and lactose. Minor decreases in levels of short chain fatty acids were observed with the cycled treatments, and the volatiles in general decreased after HP treatment, depending mostly on the pressure time but also on cycling in skimmed milk. The study confirmed the superior effect of two-cycle HP on microbial inactivation, and shows a slightly better preservation of the physical-chemical milk quality.Industrial relevanceMulti-cycling HP has been shown to be advantageous for microbial inactivation, but limited information is available regarding the effect on milk components in whole milk or skimmed milk. The present study compares the psychochemical properties of whole and skimmed milk processed by multi-cycling in comparison to single cycle HP treatment. Double cycled HP presented a superior effect on microbial inactivation and slightly better preservation of milk quality than one continuous HP.     

Impact of voltage and pulse delivery mode on the efficacy of pulsed light for the inactivation of Listeria

Listeria innocua inactivation by pulsed light (PL) was evaluated at different settings and voltages, to establish the best treatment conditions and post-treatment handling for further implementation of PL in the food industry. Fluences up to 0.2 J/cm² were applied to superficially inoculated TSA agar plates (4.5–5 log cfu/cm²). Inactivation was calculated, and log-linear and Weibull models were applied. A fluence of 0.2 J/cm² applied in a single pulse inactivated 3.8 log cfu/cm², while sequential application of this fluence yielded an inactivation between 1.5 and 2.5 log cfu/cm² depending on the delivery mode (consecutive flashing or with 5 min-holding times under ambient light or in the dark). Data from consecutive PL treatment were fitted with the Weibull model. No photoreactivation following PL was observed after 120-min exposure to ambient light in any of the conditions assayed. This study showed that flashing with a single pulse at higher voltage would offer the highest inactivation of Listeria.Industrial relevanceThis study offered information of practical interest to establish pulsed light processing and post-processing conditions for the control of Listeria spp. in the food industry, for instance in ready-to-eat (RTE) products. The use of higher voltages provided higher inactivation and allowed to minimise the number of flashes. If sequential treatments are to be applied, the treatment is more effective if short holding times are kept between pulses. The post-processing illumination conditions do not influence the efficacy of PL treatment.     

Fat content increases the lethality of ultra-high-pressure homogenization on Listeria monocytogenes in milk

Listeria monocytogenes CCUG 15526 was inoculated at a concentration of approximately 7.0 log₁₀ cfu/mL in milk samples with 0.3, 3.6, 10, and 15% fat contents. Milk samples with 0.3 and 3.6% fat content were also inoculated with a lower load of approximately 3.0 log₁₀ cfu/mL. Inoculated milk samples were subjected to a single cycle of ultra-high-pressure homogenization (UHPH) treatment at 200, 300, and 400 MPa. Microbiological analyses were performed 2 h after the UHPH treatments and after 5, 8, and 15 d of storage at 4°C. Maximum lethality values were observed in samples treated at 400 MPa with 15 and 10% fat (7.95 and 7.46 log₁₀ cfu/mL), respectively. However, in skimmed and 3.6% fat milk samples, complete inactivation was not achieved and, during the subsequent 15 d of storage at 4°C, L. monocytogenes was able to recover and replicate until achieving initial counts. In milk samples with 10 and 15% fat, L. monocytogenes recovered to the level of initial counts only in the milk samples treated at 200 MPa but not in the milk samples treated at 300 and 400 MPa. When the load of L. monocytogenes was approximately 3.0 log₁₀ cfu/mL in milk samples with 0.3 and 3.6% fat, complete inactivation was not achieved and L. monocytogenes was able to recover and grow during the subsequent cold storage. Fat content increased the maximum temperature reached during UHPH treatment; this could have contributed to the lethal effect achieved, but the amount of fat of the milk had a stronger effect than the temperature on obtaining a higher death rate of L. monocytogenes.     

Production of conjugated linoleic acid enriched milk and dairy products from cows receiving grass silage supplemented with a cereal-based concentrate containing rapeseed oil

Opportunities for the production of milk and dairy products enriched with cis-9, trans-11 conjugated linoleic acid (CLA) were investigated. Eighteen mid-lactation cows were used in a continuous-design for 7 weeks. During the first week, cows received grass silage ad libitum supplemented with 10 kg per day of a cereal-based concentrate (control) that was replaced with a concentrate containing 50 g kg⁻¹ of rapeseed oil (RO). Changes in milk fatty acid composition were monitored on a weekly basis and milk produced was used to manufacture Edam cheese and butter. Inclusion of RO in the concentrate supplement increased the mean levels of trans-octadecanoic, monounsaturated, CLA and polyunsaturated fatty acid in the milk fat from 1.6, 25.7, 0.46 and 2.8 to 4.3, 35.3, 1.02 and 3.9 g 100 g⁻¹ total fatty acids, respectively. In contrast, the mean level of saturated fatty acids decreased from 71.4 to 60.7 g  100 g⁻¹ total fatty acids. Changes in milk fatty acid composition due to RO occurred within 7 days, with responses reaching a plateau after 21 days. Furthermore, the CLA concentrations in the milk fat from individual cows ranged between 0.37 and 0.65 and 0.43 and 2.06 g 100 g⁻¹ total fatty acids for the control and RO diet, respectively. CLA enriched milk was used successfully to manufacture of Edam cheese and butter with softer textures but with acceptable organoleptic and storage properties. Processing milk into butter or cheese had no effect on the CLA concentrations indicating that enrichment of dairy products is dependent on the content in raw milk fat.     

Modeling the protective effect of aw and fat content on the high pressure resistance of Listeria monocytogenes in dry-cured ham

High pressure processing (HPP) is a promising food preservation technology as an alternative to thermal processing for microbial inactivation. The technological parameters, the type of microorganism, and the food composition can greatly affect the microbicidal potential of HPP against spoilage and pathogenic microorganisms. Presently, the number of available models quantifying the influence of food characteristics on the pathogen inactivation is scarce. The aim of this study was to model the inactivation of Listeria monocytogenes CTC1034 in dry-cured ham, as a function of pressure (347–852 MPa, 5 min/15 °C), water activity (a_w, 0.86–0.96) and fat content (10–50%) according to a Central Composite Design. The response surface methodology, based on the equation obtained with a stepwise multivariate linear regression, was used to describe the relationship between bacterial inactivation and the studied variables. According to the best fitting polynomial equation, besides pressure intensity, both a_w and fat content exerted a significant influence on HP-inactivation of L. monocytogenes. A clear linear piezoprotection trend was found lowering the a_w of the substrate within the whole range of tested pressure. Fat content was included in the model through the quadratic term and as interaction term with pressure, resulting in a particular behavior. A protective effect due to the presence of high fat content was identified for pressure treatments above ca. 700 MPa. At lower pressure, higher inactivation of L. monocytogenes occurred by increasing the fat content above 30%. The results emphasize the relevant influence of intrinsic factors on the L. monocytogenes inactivation by HPP, making necessary to assess and validate the effectiveness of HPP on specific food products and consequently set process criteria adjusted to each particular food product.     

A microbiological perspective of raw milk preserved at room temperature using hyperbaric storage compared to refrigerated storage

The effects of hyperbaric storage (HS, 50–100 MPa) at room temperature (RT) on endogenous and inoculated pathogenic surrogate vegetative bacteria (Escherichia coli, Listeria innocua), pathogenic Salmonella enterica and bacterial spores (Bacillus subtilis) were assessed and compared with conventional refrigeration at atmospheric pressure for 60 days. Milk stored at atmospheric pressure and refrigeration quickly surpassed the acceptable microbiological limit within 7 days of storage, regarding endogenous microbiota, yet 50 MPa/RT slowed down microbial growth, resulting in raw milk spoilage after 28 days, while a significant microbial inactivation occurred under 75–100 MPa (around 4 log units), to counts below 1 log CFU/mL throughout storage, similar to what was observed for B. subtilis endospores. While inoculated microorganisms had a gradually counts reduction in all HS conditions. Results indicate that HS can not only result in the extension of milk shelf-life but is also able to enhance its safety and subsequent quality.Industrial relevanceThis new preservation methodology could be implemented in the dairy farm storage tanks, or during milk transportation for further processing, allowing a better microbial control, than refrigeration. This methodology is very promising, and can improve food products shelf-life with a considerable lower carbon foot-print than refrigeration.     

Milk colloidal system as a reaction medium and carrier for the natural blue colorant obtained from the cross-linking between genipin and milk proteins

A promising natural blue colorant was obtained from the cross-linking between genipin and milk proteins. Milk was simultaneously used to extract genipin from the unripe genipap (Genipa americana L.) and evaluated as a reaction medium and carrier for the novel blue colorant. The effects of the milk composition (skimmed, semi-skimmed, and whole) on the kinetic of blue color formation in the colorant-loaded milk samples during their cold storage time for 96 h were evaluated using their color parameters and free-genipin content. The reaction between milk proteins and genipin were evaluated by FTIR spectroscopy. In addition, the blue colorant-loaded milk samples were characterized according to their droplet size distribution, microstructure, and phase separation kinetics. The milk fat content influenced the genipin recovery while the milk with higher protein content contributed to obtaining a more intense blue color. Using whole milk with 3.0 g/100 g fat content favored the light scattering and, thus, a blue colorant with a more intense −b¹ and L¹ values was obtained. On the other hand, the use of skimmed milk with 0.5 g/100 g fat content resulted in more consumption of genipin due to its more protein content for the reaction of blue color compounds formation. Thus, a more intense and darker blue coloration was observed with lower light scattering. The milk composition did not modify the phase separation kinetics of the blue colorant-loaded milk. Therefore, our results have demonstrated that the milk was a suitable medium for the reaction of blue color formation and also a good blue compounds carrier.Industrial relevanceConsumer demands for natural colorants have increased in the last years.Thus, the new food industry challenge is to develop novel healthy, safe and high-quality food products based on natural colorants. Currently, the colorant market does not have still available a natural blue colorant. Therefore, the development of a novel natural blue colorant from plant material could meet this worldwide demand increasing the added value of many products such as ice cream, dairy beverages, and candies.     

Microbial quality of raw horse milk

Consumption of horse milk has become popular in developed countries, especially among people suffering from bowel problems and skin diseases. Since the positive effect is supposedly not observed after pasteurisation, the product is mostly consumed as raw milk. Since the microbiological quality of this milk has not been systematically surveyed, in this study we examined the presence of spoilage- and pathogenic microorganisms in 123 samples of horse milk collected in The Netherlands and Belgium. Hygiene and faecal indicators were found in a wide range of numbers. Although Salmonella, Campylobacter and Listeria were not found, these pathogens showed no reduction in challenge tests with artificially contaminated horse milk stored for 1 week at 7 °C. Since faecal indicators were present and able to grow at 7 °C, combined with the fact that pathogens may easily end up and survive in the milk, it is not advised to consume raw horse milk.     

Kinetic models for pulsed electric field and thermal inactivation of Escherichia coli and Pseudomonas fluorescens in whole milk

Inactivation of Escherichia coli ATCC 11775 and Pseudomonas fluorescens ATCC 948 in UHT whole (4% fat) milk during thermal processing at 56–62 °C and pulsed electric field (PEF) processing at 30 or 35 kV cm⁻¹ at approximately 30, 40 or 50 °C was investigated. E. coli ATCC 11775 was more heat-resistant than P. fluorescens ATCC 948, but more susceptible to PEF processing. All inactivation kinetics showed strong deviations from log-linearity. Thus, a simplified logistic (log-decay) regression model was used to accurately predict thermal and PEF inactivation of E. coli ATCC 11775 and P. fluorescens ATCC 948 under various treatment conditions. This is a useful tool for identifying processing conditions to inactivate pathogenic and spoilage microorganisms in whole milk at sub-pasteurisation temperatures.     

Effects of ultrasound on the fermentation profile and metabolic activity of lactic acid bacteria in buffalo's (Bubalus bubalis) milk

In this study, the ultrasound was applied at the lag phase and logarithmic phase of the growth of Streptococcus thermophilus subsp. Salivarius (6.76 ± 0.15 log cfu/mL) and Lactobacillus bulgaricus (6.36 ± 0.22 log cfu/mL) in buffalo's milk. The results revealed that S. thermophilus entered into the logarithmic phase after 45 min of incubation. Ultrasound application during lag phase (1685 J mL⁻¹) and logarithmic phase (561.6 J mL⁻¹) reduced the fermentation time by 32 min (12.5%) and 40 min (15.7%), respectively. Ultrasound treatment (1684.8 J mL^-1) during the lag phase resulted in fractures (approximately 0.1–0.2 μm) on the cell membrane of S. thermophilus. Application of ultrasound for a 2–6 min period enhanced (P < 0.05) the microbial growth while increasing (P < 0.05) the β-galactosidase activity. The lactose hydrolysis was increased up to 49.2% compared to that of the control. Ultrasound treatment during the lag phase showed higher (P < 0.05) storage modulus, yield point and consistency index compared to the logarithmic phase. It can be concluded that ultrasound has huge potential to improve the fermentation rate of buffalo's milk.Industrial relevanceConventional food fermentation is one of the most time and resources consuming processing steps in the food industry. In this work, it was shown that the appropriate application of ultrasound during different stages of microbial growth reduced the fermentation time of buffalo milk by 13% to 16% by enhancing the metabolic activities of lactic acid bacteria. Therefore, ultrasound has great prospects in the food processing industry to improve process efficiency. Scaling-up of laboratory applications of US in to industrial scale through designing new power transducers with high capacity and greater acoustic filtration is necessary to enhance its potential to be used at large scales.     

Microstructure of fat globules in whole milk after thermosonication treatment

The structure of fat globules in whole milk was studied after heat and thermosonication treatments to observe what happens during these processes at the microscopic level using scanning electron microscopy. Raw whole milk was thermosonicated in an ultrasonic processor-Hielscher UP400S (400 W, 24 kHz, 120 microm amplitude), using a 22-mm probe at 63 degrees C for 30 min. Heat treatment involved heating the milk at 63 degrees C for 30 min. Color and fat content were measured to correlate the images with analytical measurements. The results showed that the surface of the fat globule was completely roughened after thermosonication. Ultrasound waves were responsible for disintegrating the milk fat globule membrane (MFGM) by releasing the triacylglycerols. Furthermore, the overall structure of milk after sonication showed smaller fat globules (smaller than 1 microm) and a granular surface. This was due to the interaction between the disrupted MFGM and some casein micelles. Minor changes in the aspect of the globules between thermal and raw milks were detected. Color measurements showed higher L* values for sonicated samples. Sonicated milk was whiter (92.37 +/- 0.20) and generally showed a better degree of luminosity and homogenization compared to thermal treated milk (88.25 +/- 0.67) and raw milk (87.82 +/- 0.18). Fat content analysis yielded a higher value after sonication (4.24%) compared to untreated raw milk (4.04%) because fat extraction is more efficient after sonication. The advantages of thermosonicated milk are that it can be pasteurized and homogenized in just 1 step, it can be produced with important cost savings, and it has better characteristics, making thermosonication a potential processing method for milk and most other dairy products.     

Factors affecting 3D printing and post-processing capacity of cookie dough

This study aimed to investigate the factors affecting the printability and post-processing capacity of cookie dough in extrusion-based three-dimensional (3D) printing by modifying the recipe without the addition of gums or stabilizers. Cookie dough formulations with different types of fat (butter and shortening), flour (wheat, rice, and tapioca), the amount of non-fat milk (32.5 or 65 g/100 g flour), and the sugar level (37.5 or 55 g/100 g flour) were investigated for their printability and post-processing capacity. Rheological properties, microstructure, and printability of printing inks were monitored while the moisture loss and dimensional stability after baking were analyzed in printed and baked structures, respectively. Results indicated that cookie dough formulations with reduced sugar content were more printable. The best sample to build a shape that could withstand the baking process was the recipe of 37.5 g sugar, 62.5 g shortening, 100 g tapioca flour, and 32.5 g milk, which yielded printed cookie samples with the ease of printing, better visual printing outcomes, and no structural deformation after baking.Industrial relevance3D food printing is an emerging technology with many potential applications in the food industry. It is important to understand the effects of key components of food materials on the printing, which enable a wider range of structures just using a single nozzle, and provide tailored nutrition and personalization in extrusion-based 3D printing. Understanding the effects of food processing on 3D printed food items is critical to broadening its applications. This study selected cookie dough as a model system to investigate the 3D food printing since it is mainly composed of fat, sugar, flour, and milk that are the most common ingredients in many food products. Establishing knowledge of the interactions between different food components and contribution of each individual ingredient may help develop a guideline for extrusion-based 3D food printing. In addition, the results acquired from this study can fill knowledge gaps regarding the role of cookie dough component in 3D printing and their effects on post-processing of 3D printed foods.     

Characterization of metastable high pressure phase transition positions and its influence on the behavior of microbial destruction

Effect of perturbation factors on phase transition metastable positions of whole milk (4% fat content) and their influence on microbial destruction characteristics of non-pathogenic Escherichia coli inoculated in milk subjected to high pressure low temperature treatment were evaluated using a specially developed high pressure (HP) cooling system. Initially, the phase transition data of milk transitioning through the metastable phases were obtained and fitted successfully using Simon-like models as done in previous studies and polynomial formulas with R² of 0.997 & 0.996 for ice I, and 0.989 & 0.989 for ice III, respectively. The phase transition position of milk was explored with 5% and 10% sodium chloride solution as perturbation sources, respectively. Results showed that the 5% sodium chloride solution can reduce the transition pressure of milk by 43 MPa and increase the transition temperature by 4.1 °C, so that the milk can achieve phase transition at lower pressure and higher temperature. Phase transition microbial destruction was characterized by discontinuity, mutation and segmentation when the phase transition pressure interval 250– 300 MPa was carefully refined. The inactivation amount of E. coli before the phase transition (250 MPa) was 1.11 log and the phase transition process itself brought an additional 1.26 log destruction of E. coli population in milk.Industrial relevanceHigh pressure low temperature (HPLT) phase change kinetics were employed to enhance microbial destruction. HPLT was established based on a self-cooling unit positioned inside conventional HP chamber offering opportunities for scale up and commercialization. The effectiveness of HPLT phase transition for Escherichia coli destruction was demonstrated. The related research in metastable state provides a reference point for commercial application of high-pressure-low-temperature technology for microbial destruction and quality enhancement.     

Kinetic and thermodynamic study of thermal inactivation of the antimicrobial peptide P34 in milk

The knowledge on thermal inactivation of biopreservatives in a food matrix is essential to allow their proper utilisation in food industry, enabling the reduction of heating times and optimisation of heating temperatures. In this work, thermal inactivation of the antimicrobial peptide P34 in skimmed and fat milk was kinetically investigated within the temperature range of 90–120 °C. The inactivation kinetic follows a first-order reaction with k-values between 0.071 and 0.007 min⁻¹ in skimmed milk, and 0.1346 and 0.0119 min⁻¹ in fat milk. At high temperatures, peptide P34 was less resistant in fat milk, with a significant decrease in residual activity as compared with skimmed milk. At temperatures below 110 °C, the fat globules seem to have protective effect to the peptide P34. Results suggest that peptide P34 is heat stable in milk with activation energy of 90 kJ mol⁻¹ in skimmed milk and 136 kJ mol⁻¹ in fat milk.     

Addition of olein from milk fat positively affects the firmness of butter

Butter is highly valued for its characteristic flavor and aroma; however, it has the disadvantage of unsatisfactory spreadability at low temperatures. The functional characteristics of butter can be modified by changing its composition or physical structure. The objective of this study was to evaluate the effect of olein on structure and composition of butter. Olein was obtained by two stage dry fractionation process of the anhydrous milk fat (AMF) and added to commercial cream prior to butter manufacture. The fractions were characterized for triacylglycerols composition, solid fat content, crystallization isotherm, and thermal behavior. Butter was manufactured using commercial cream or cream containing 50% olein. Butter samples were characterized for physicochemical composition, instrumental color, crystallization parameters, and firmness after 1 and 7 days of storage at 10 °C. The firmness of butter subjected to room temperature was also evaluated. Butter containing olein differed significantly from the control and had darker yellow color, higher crystallization time, and lower solid fat content after 120 min at 15 °C, and hence lower firmness after 1 and 7 days of refrigerated storage. Although lower firmness was observed over time for all samples at room temperature, butter containing olein exhibited lower firmness after both 1 and 7 days, thus suggesting changes in organization of solid fat crystal network in the liquid fat. The addition of olein to butter allowed obtaining a softer product, with more intense color and possible nutritional benefits due to the medium chain triglycerides and higher carotene levels.     

Development and evaluation of a processing sector model for butter manufacture using a mass balance technique at two dairy processing sites

The butter manufacturing process at two different commercial dairy processing sites in Ireland was evaluated using a mass balance approach to develop, evaluate and validate a processing sector model of the flow of milk fat from intake to final product. The mass balance was represented as a function of fat intake = fat in products + fat losses + recycled fat. Representative samples of all products, namely whole milk, cream, skim milk, butter, buttermilk and cleaning‐in‐place streams (cream silo flush, butter churn residue and sludge), were collected from two different sites. Milk fat levels and product quantities were measured to obtain the fat outputs. Total fat losses at the end of butter production ranged between 1.90% and 2.25% of the total fat input for both sites. Three different scenarios were examined to evaluate the model: S1 (Animal Breed) high genetic merit (Elite) and national average (NA) Holstein Friesian (HF) cows were evaluated, for their effect on the net value of milk; S2 (Product Portfolio) a mixed product portfolio of cheese, butter and skim milk powder (SMP) was compared to a product portfolio comprised of butter alone; and S3 (Process Efficiency) the impact of varying process losses on net values of milk and the quantities of products produced was simulated. The value per 1000 L of milk for S1 was €410.69 and €393.20 for Elite and NA cow’s milk, respectively. For S2, the butter‐only product portfolio returned €355.10, whereas the mixed‐products portfolio returned €369.60. Lastly, S3 corresponding returns for 1%, 2.2% and 5% losses was €365.90, €361.47 and €351.12, respectively.     

Enrichment of the conjugated linoleic acid content of bovine milk fat by dry fractionation

This study investigated the effect of fat fractionation on the conjugated linoleic acid (cis-9, trans-11-C_18 : 2) content of bovine milk fat. Anhydrous milk fat was fractionated into hard and soft fractions using controlled cooling and agitation. Fractionation of milk fat pre-melted at 60°C using a temperature programme of 33–10°C and a cooling rate of 0.58°C h⁻¹ yielded a soft fraction containing 63.2% more conjugated linoleic acid (2.22 g 100 g⁻¹ FAME), which was also enriched in polyunsaturated fatty acids and vaccenic acid (trans-11-C_18 : 1) compared with the parent fat. Agitation following fractionation was found to have a negative effect on the conjugated linoleic acid content of the soft fraction. Refractionation of the soft fraction did not increase the yield of conjugated linoleic acid. The conjugated linoleic acid and trans fatty acid content of 26 selected food products ranging in milk fat content from 0 to 100% is reported. Conjugated linoleic acid concentrations ranged from 0 to 16.2 mg g⁻¹ fat and were generally lower than the trans fatty acid content which ranged from 0 to 155.7 mg g⁻¹ fat. Spreads containing vegetable oils contained higher trans fatty acid and lower conjugated linoleic acid contents than milk fat-containing products. This study highlights that a milk fat fraction enriched in conjugated linoleic acid may be achieved by dry fractionation.     

Protease stability in bovine milk under combined thermal-high hydrostatic pressure treatment

At atmospheric pressure, inactivation of protease from B. subtilis in raw milk and pasteurized milk (with and without homogenization) was studied in a temperature range of 50–80 °C. Thermal inactivation followed a first order kinetic model in the temperature range tested. Temperature dependence of the first order inactivation rate constants could be accurately described by the Arrhenius equation, allowing Ea values to be calculated. Different milk systems did not show differences in enzyme thermo stability.The combined thermal (40, 50 and 60 °C)-high hydrostatic pressure (300–450 and 600 MPa) effect on protease activity was studied. Protease was very resistant to high pressures. Pressure stability was higher in raw milk than in pasteurized milk; homogenization appeared to have a protective effect on the enzyme. The separate effects of pressure and temperature on enzyme inactivation were related to changes in L?-values and milk appearance.A very pronounced antagonistic effect between high temperature and pressure was observed, i.e. at temperatures where thermal inactivation at atmospheric pressure occurs rapidly, application of pressure up to 600 MPa exerted a protective effect.Industrial relevanceHigh hydrostatic pressure (HHP) is an emerging technology that has been successfully applied as a minimal process for a variety of foods. Although the potential for the use of HHP treatment as an alternative method to heat treatment of milk was proposed almost a century ago, the suitability of this innovative technology to extend the shelf-life of milk hinges not only on its ability to inactivate pathogenic vegetative microorganisms but also on its effectiveness to inactivate indigenous and endogenous enzymes. This work examines the combined effects of temperature, pressure and homogenization on the protease (exogenous enzyme from B. subtilis) activity in milk. Inactivation of protease could extend the shelf life of milk.