Ionic strength and buffering capacity of emulsifying salts determine denaturation and gelation temperatures of whey proteins.

Ionic conditions affect the denaturation and gelling of whey proteins, affecting the physical properties of foods in which proteins are used as ingredients. We comprehensively investigated the effect of the presence of commonly used emulsifying salts on the denaturation and gelling properties of concentrated solutions of ß-lactoglobulin (ß-LG) and whey protein isolate (WPI). The denaturation temperature in water was 73.5°C [coefficient of variation (CV) 0.49%], 71.8°C (CV 0.38%), and 69.9°C (CV 0.41%) for ß-LG (14% wt/wt), ß-LG (30% wt/wt), and WPI (30% wt/wt), respectively. Increasing the concentration of salts, except for sodium hexametaphosphate, resulted in a linear increase in the denaturation temperature of WPI (kosmotropic behavior) and an acceleration in its gelling rate. Sodium chloride and tartrate salts exhibited the strongest effect in protecting WPI against thermal denaturation. Despite the constant initial pH of all solutions, salts having buffering capacity (e.g., phosphate and citrate salts) prevented a decrease in pH as the temperature increased above 70°C, resulting in a decline in denaturation temperature at low salt concentrations (≤0.2 mol/g). When pH was kept constant at denaturation temperature, all salts except sodium hexametaphosphate, which exhibited chaotropic behavior, exhibited similar effects on denaturation temperature. At low salt concentration, gelation was the controlling step, occurring up to 10°C above denaturation temperature. At high salt concentration (>3% wt/wt), thermal denaturation was the controlling step, with gelation occurring immediately after. These results indicate that the ionic and buffering properties of salts added to milk will determine the native versus denatured state and gelation of whey proteins in systems subjected to high temperature, short time processing (72°C for 15 s).

High-pressure homogenization and high hydrostatic pressure processing of human milk: Preservation of immunological components for human milk banks.

Human milk (HM) constitutes the first immunological barrier and the main source of nutrients and bioactive components for newborns. Immune factors comprise up to 10% of the protein content in HM, where antibodies are the major components (mainly IgA, IgG, and IgM). In addition, antibacterial enzymes such as lysozyme and immunoregulatory factors such as soluble cluster of differentiation 14 (sCD14) and transforming growth factor ß2 (TGF-ß2) are also present and play important roles in the protection of the infant's health. Donor milk processed in HM banks by Holder pasteurization (HoP; 62.5°C, 30 min) is a safe and valuable resource for preterm newborns that are hospitalized, but is reduced in major immunological components due to thermal inactivation. We hypothesized that high hydrostatic pressure (HHP) and high-pressure homogenization (HPH) are 2 processes that can be used on HM to reduce total bacteria counts while retaining immunological components. We studied the effects of HHP (400, 450, and 500 MPa for 5 min applied at 20°C) and HPH (200, 250, and 300 MPa, milk inlet temperature of 20°C) applied to mature HM, on microbiological and immunological markers (IgA, IgG, IgM, sCD14, and TGF-ß2), and compared them with those of traditional HoP in HM samples from healthy donors. The HHP processing between 400 and 500 MPa at 20°C reduced counts of coliform and total aerobic bacteria to undetectable levels (<1.0 log cfu/mL) while achieving approximately 100% of immunological component retention. In particular, comparing median percentages of retention of immunological components for 450 MPa versus HoP, we found 101.5 versus 50.5% for IgA, 89.5 versus 26.0% for IgM, 104.5 versus 75.5% for IgG, 125.0 versus 72.5% for lysozyme, 50.6 versus 0.1% for sCD14, and 88.5 versus 61.1% for TGF-ß2, respectively. Regarding HPH processing, at a pressure of 250 MPa and inlet temperature of 20°C, the process showed good potential to reduce coliforms to undetectable levels and total aerobic bacteria to levels slightly above those obtained by HoP. The median percentages of retention of immunological markers for HPH versus HoP were 71.5 versus 52.0%, 71.0 versus 27.0%, 104.0 versus 66.5%, and 30.9 versus 0.2%, for IgA, IgM, IgG, and sCD14, respectively; results did not significantly differ for lysozyme and TGF-ß2. The HPH at 300 MPa produced higher inactivation of immunological components, similar to values achieved with HoP.

Milk fatty acid profile from grass feeding strategies on 2 Holstein genotypes: Implications for health and technological properties.

The objective of the study was to determine if a feeding system with a variable supply of grass promoted rapid changes in the fatty acid profile and technological and health indices of milk obtained from North American (NAHF) and New Zealand (NZHF) Holstein-Friesian cows. Two feeding strategies were conducted: fixed grass (GFix) and maximized grass intake when available (GMax). The results showed that as the grass intake increased in the GMax treatments, the relative amount of palmitic acid in milk decreased, whereas oleic, linoleic, linolenic, and conjugated linoleic acids increased, causing a reduction in the atherogenic, thrombogenic, and spreadability calculated indices. The changes occurred in rapid response to the changing diet, with reductions ranging from approximately 5 to 15% in the healthy and technological indices within a period of 15 d of grass intake increase. Differences were found between the 2 genotypes, with NZHF responding faster to changes in grass intake.

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 log(10) 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 log(10) 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 degrees C. Maximum lethality values were observed in samples treated at 400 MPa with 15 and 10% fat (7.95 and 7.46 log(10) 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 degrees 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 log(10) 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.

Inactivation of Salmonella enterica serovar Senftenberg 775W in liquid whole egg by ultrahigh pressure homogenization.

Two batches of samples of liquid whole egg were inoculated with a load of approximately 3 and 7 log CFU/ml, respectively, of Salmonella enterica serovar Senftenberg 775W and submitted to different ultrahigh pressure homogenization (UHPH) treatments at 150, 200, and 250 MPa. The inlet temperature of the samples was 6 degrees C. Counts of viable and injured Salmonella cells were obtained 2 h after the UHPH treatments and after 5, 10, 15, and 20 days of storage at 4 degrees C. The level of pressure applied influenced the lethality attained significantly (P < 0.05). In the samples with an initial load of approximately 7 log CFU/ml, the highest lethality value of 3.2 log CFU/ml was obtained at 250 MPa, and it is similar to those values reported in other surveys for thermal pasteurization with this same Salmonella strain. When the initial load was approximately 3 log CFU/ml, total inactivation was apparently obtained after the 250-MPa treatment (2.7 log CFU/ml). After 10 days of storage at 4 degrees C, Salmonella counts decreased in UHPH-treated samples, and colonies were not observed in tryptone soy agar and yeast extract medium. Nevertheless, presence of viable Salmonella cells was detected with the VIDAS Salmonella immunoassay method during the entire storage period. These results encourage further investigation of UHPH processing of liquid whole egg, assaying the possibility of using higher pressures and fluid inlet temperatures.

Response of two Salmonella enterica strains inoculated in model cheese treated with high hydrostatic pressure.

The aim of this work was to determine the response to high hydrostatic pressure and the ability for survival, recovery, and growth of 2 strains of Salmonella enterica (Salmonella enteritidis and Salmonella typhimurium) inoculated in a washed-curd model cheese produced with and without starter culture. Inoculated samples were treated at 300 and 400 MPa for 10 min at room temperature and analyzed after treatment and after 1, 7, and 15 d of storage at 12 degrees C to study the behavior of the Salmonella population. Cheese samples produced with starter culture and treated at 300 and 400 MPa showed maximum lethality; no significant differences in the baroresistant behavior of both strains were detected. Nevertheless, when starter culture was not present, the maximum lethality was only observed in cheese samples treated at 400 MPa, in the case of S. enteritidis. Ability to repair and grow was not observed in model cheese produced with starter culture and cell counts of treated samples decreased after 15 d of storage at 12 degrees C. In cheese produced without starter culture, Salmonella cells showed the ability to repair and grow during the storage period, reaching counts over 3 log(10) (cfu/mL) in both applied treatments and serotypes. These results suggest that high hydrostatic pressure treatments are effective to reduce Salmonella population in this type of cheese, but the presence of the starter culture affects the ability of this microorganism to repair and grow during the storage period.

Fate of Staphylococcus aureus in cheese treated by ultrahigh pressure homogenization and high hydrostatic pressure.

We evaluated the influence of ultrahigh pressure homogenization (UHPH) treatment applied to milk containing Staphylococcus aureus CECT 976 before cheese making, and the benefit of applying a further high hydrostatic pressure (HHP) treatment to cheese. The evolution of Staph. aureus counts during 30 d of storage at 8 degrees C and the formation of staphylococcal enterotoxins were also assessed. Milk containing approximately 7.3 log(10) cfu/mL of Staph. aureus was pressurized using a 2-valve UHPH machine, applying 330 and 30 MPa at the primary and the secondary homogenizing valves, respectively. Milk inlet temperatures (T(in)) of 6 and 20 degrees C were assayed. Milk was used to elaborate soft-curd cheeses (UHPH cheese), some of which were additionally submitted to 10-min HHP treatments of 400 MPa at 20 degrees C (UHPH+HHP cheese). Counts of Staph. aureus were measured on d 1 (24 h after manufacture or immediately after HHP treatment) and after 2, 15, and 30 d of ripening at 8 degrees C. Counts of control cheeses not pressure-treated were approximately 8.5 log(10) cfu/g showing no significant decreases during storage. In cheeses made from UHPH treated milk at T(in) of 6 degrees C, counts of Staph. aureus were 5.0 +/- 0.3 log(10) cfu/g at d 1; they decreased significantly to 2.8 +/- 0.2 log(10) cfu/g on d 15, and were below the detection limit (1 log(10) cfu/g) after 30 d of storage. The use of an additional HHP treatment had a synergistic effect, increasing reductions up to 7.0 +/- 0.3 log(10) cfu/g from d 1. However, for both UHPH and UHPH+HHP cheeses in the 6 degrees C T(in) samples, viable Staph. aureus cells were still recovered. For samples of the 20 degrees C T(in) group, complete inactivation of Staph. aureus was reached after 15 d of storage for both UHPH and UHPH+HHP cheese. Staphylococcal enterotoxins were found in controls but not in UHPH or UHPH+HHP treated samples. This study shows a new approach for significantly improving cheese safety by means of using UHPH or its combination with HHP.