This research communication describes enzymatic and physico-chemical changes during storage of UHT milk. The UHT milk sample was stored at 5 and 30°C for 4 months and analyzed regularly at intervals of one month. During storage of UHT milk, there was a significant (P < 0.001) increase in non-protein nitrogen, non-casein nitrogen, soluble calcium, soluble magnesium and proteolysis, while a significant (P < 0.001) decrease in pH was observed. There was a slight change in the particle size and zeta potential of casein micelles. Changes were more pronounced in milk samples stored at 30°C than in those stored at 5°C. During storage, there occurred changes in pH, viscosity, salt balance and nitrogenous components which adversely affected quality. It was concluded that the proteolysis led to the acidification which had a destabilizing effect on the milk.

Porungo is a traditional pasta filata cheese produced using raw milk throughout the southwest region of the state of São Paulo, Brazil. The objectives of this Research Communication were to evaluate the quality of raw milk used to make Porungo cheese, to characterize its production process, and to determine its chemical composition. The results showed that the raw milk met both Brazilian and international quality requirements. Chemically, Porungo can be classified as a medium to full fat semi-hard fresh cheese. Our study has allowed the first standards and regulations for Porungo to be established in Brazil. By virtue of this, the local producers are able to formalize their activity while consumers can have access to a safe and certified product.

The objectives of this study were: to assess the efficiency of high hydrostatic pressure or ultra-high pressure homogenization against Mycobacterium smegmatis in milk and to discuss whether M. smegmatis can be considered a suitable surrogate for other Mycobacterium spp. in high pressure inactivation trials using milk. Three strains of this specie (CECT 3017, 3020 and 3032) were independently inoculated into both skimmed (0.2% fat) and whole milk (3.4% fat) at an approximate load of 6.5 Log CFU/ml and submitted to HHP treatments at 300, 400 or 500 MPa for 10 m at 6°C and 20°C. Evolution of the surviving cells of the inoculated strains was evaluated analysing milk immediately after the treatments and after 5 and 8 d of storage at 6°C. HHP treatments at 300 MPa were seldom efficient at inactivating M. smegmatis strains, but lethality increased with pressure applied in all cases. Generation of sub-lethal injured cells was observed only after 400 MPa treatments since inactivation at 500 MPa was shown to be complete. Significant differences were not observed due to either temperature of treatment or fat content of milk, except for strain CECT3032, which was shown to be the most sensitive to HHP treatments. Milk inoculated with strain CECT3017 was submitted to ultra-high pressure homogenization (UHPH) treatments at 200, 300 and 400 MPa. Maximum reductions were obtained after 300 and 400 MPa treatments, although less than 3.50 Log CFU/ml were inactivated. UHPH did not cause significant number of injured cells. The usefulness of this species as a marker for pressure-based processing seems limited since it showed greater sensitivity than some pathogenic species including other Mycobacteria reported in previous studies.

The relationship between the cow and calf develops over time after birth. The behavioural mechanisms underlying its development are important and comparisons with other species may increase our understanding. In nature the cow will separate herself from the herd to give birth and then the cow–calf relationship will develop with the ability to recognise each other. While twinning levels are low in cows, they do rear their twin calves. If the calf is lost at or after birth the cow can be responsive towards other calves and in specific circumstances the cow can develop a maternal bond with an alien calf, i.e. foster. In this Research Reflection a distinction is made between the development of, on the one hand, maternal responsiveness (the tendency of the cow to care for a calf which occurs before birth) and, on the other hand, the development of the maternal–filial bond or relationship which is reciprocal, occurs after birth and is characterised by the ability to discriminate the mother's own calf from alien calves. These processes can overlap and the relationship between cow and calf in this ‘hider’ species is more plastic than in some other mammals. For example, a cow might form an attachment with an alien calf before she gives birth. After the cow has given birth the loss of her own calf may result in the state of maternal responsiveness being maintained, such that developing a maternal bond with one or several appropriate alien calves is possible. Viable fostering techniques are possible. If a maternal relationship to the mother's own calf has developed then fostering will be more difficult. If the cow's relationship with her own calf is not exclusive, and she is in a state of maternal responsiveness then fostering of calves of an appropriate age and status can be achieved.

Antioxidant components of colostrum prevent oxidative cell damage caused by free radicals that could harm the calf's development. The relationship of antioxidant potential of colostrum with parity is not well defined and could enlighten the importance of these components for the neonate and for the protection of the intestinal epithelium. The purpose of this work was to determine the antioxidant potential of colostrum from primiparous and multiparous Holstein cows in a commercial dairy farm. Samples from the first milk secretion from primiparous (first lactation, n = 8) and multiparous (second and third lactations, n = 8) Holstein cows were collected after birth of calves for determination of immune and antioxidant factors. The cows sampled in this study were vaccinated during pregnancy in order to improve colostrum quality. Colostrum from primiparous cows showed higher values of ceruloplasmin activity, oxygen radical absorbance capacity (ORAC) and transferrin saturation index (TSI) than colostrum from multiparous cows (P < 0.05). The total iron binding capacity (TIBC) and transferrin concentration in the colostrum of primiparous cows showed a non-significant numerical decrease (P = 0.06) in relation to the value in the colostrum of multiparous cows. Concentration of proteins, immunoglobulin G, and activity of lactoperoxidase, lysozyme, glutathione peroxidase and catalase, in turn, did not differ (P > 0.05). Metabolic differences between primiparous and multiparous cows may have affected the antioxidative status of colostrum, since ORAC values were twice higher in first lactation cows. Lower values of transferrin and TIBC and higher TSI in colostrum from primiparous cows suggests a relationship between lower iron stock and higher antioxidant activity. Thus, this work indicates an important role of the antioxidant potential of colostrum for neonates from first-lactation cows. Additionally, the iron stock may be directly related to the higher antioxidant potential of the colostrum from primiparous cows, and further investigations are required.

We compared the electrical conductivity from two different aggregates of whey protein concentrates (WPC) film: conventional amorphous aggregation at natural pH (pH 6.5) and amyloid fibrils at a low pH (pH 2.0) far away from the isoelectric point. The two types of film fabricated by these solutions with different aggregate structures showed large variations in electrical conductivity and other properties. The WPC fibril film (pH 2.0) exhibited higher electrical conductivity than that of the conventional WPC film (pH 6.5), improved mechanical properties and oil resistance, due to varying morphology, higher surface hydrophobicity and more (absolute value) surface charge of film-forming solutions. The evidence from this study suggests that fibrilized WPC with high-ordered and β-sheets-rich structures fabricated high electrical conductivity film, which broadens the potential application of fibrils as functional bio-nanomaterials.

Due to increasing public concern regarding separation of the dairy cow and calf within the first days after birth, alternative systems, where cows and calves stay in contact for an extended period, are receiving increasing interest from a broad array of researchers and other stakeholders. With more research in the area, there is a risk of inconsistencies emerging in the use of terminology. To create a better consensus in further discussions, the aim of this Research Reflection is to provide definitions and propose a common terminology for cow-calf contact in dairy production. We also suggest definitions for various systems allowing cow-calf contact and describe the distinct phases of cow-calf contact systems.

For this research communication, 90 samples of a Brazilian dairy were combined into four groups (raw material, final product, food-contact and non-food contact surfaces) and analyzed by metataxonomics based on 16S rRNA gene sequencing. The results showed high alpha-diversity indexes for final product and non-food contact surfaces but, overall, beta-diversity indexes were low. The samples were separated in two main clusters, and the core microbiota was composed by Macrococcus, Alkaliphilus, Vagococcus, Lactobacillus, Marinilactibacillus, Streptococcus, Lysinibacillus, Staphylococcus, Clostridium, Halomonas, Lactococcus, Enterococcus, Bacillus and Psychrobacter. These results highlight that rare taxa occur in dairies, and this may aid the development of strategies for food protection.

Using membrane filtration it is possible to selectively concentrate proteins and, in the case of microfiltration, concentrate casein micelles. During filtration, water is often added and this practice, called diafiltration, causes further release of permeable components and maintains filtration efficiency. Filtration causes changes in composition of the protein as well as the soluble phase, including soluble calcium, which is a critical factor controlling the gelation properties of the casein micelles in milk. It was hypothesized that concentrates obtained using membrane filtration with or without diafiltration would have different gelation behavior. To test this hypothesis, two concentrates of similar casein micelle volume fraction were prepared, using spiral wound polymeric microfiltration membranes with a 800 kDa molecular weight cutoff, with or without diafiltration. The concentrates showed a gelation behavior comparable to that of skim milk, with a similar gelation time and with a higher firmness, due to the higher number of protein linkages in the network. In contrast, the hydrolysis of κ-casein by chymosin and casein aggregation were inhibited in diafiltered casein micelle suspensions. When the concentrates were recombined with the original skim milk to a final concentration of 5% protein, which re-established a similar soluble phase composition, differences in gelation behavior were no longer observed: both treatments showed similar gelation time and gel firmness. These results confirmed that membrane filtration can result in concentrates with different functionality, and that ionic environmental conditions are critical to the aggregation behavior of casein micelles. This is of particular significance in industrial settings where these fractions are used as a way to standardize proteins in cheese making.

This research communication relates to the hypothesis that the consumption of raw or unprocessed cow's milk contributes to lowered prevalence of allergies. Thermal pasteurization of bovine milk can result in denaturation of minor whey proteins and loss of their bioactivity. Denaturation of bovine serum albumin (BSA), immunoglobulin G (IgG) and lactoferrin (LF) in skim milk was studied under different temperature (72, 75 or 78°C) and time (0–300 s) combinations. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) results revealed that denaturation of all 3 proteins occurred at 72°C and progressed with increase in temperature and holding time. About 59% of LF and 12% of IgG denatured under high-temperature short-time (72°C/ 15 s) pasteurization, while BSA was least impacted. To assess modulation of milk immunogenicity, secretion of selected T helper (Th)-type cytokines by human peripheral blood mononuclear cells (PBMCs) was studied in vitro in response to different concentrations of BSA (0.4–1.0 mg/ml) and IgG (0.8–1.6 mg/ml) in unheated skim milk. Addition of IgG at 1.6 mg/ml induced a prominent Th1-skewed cytokine profile that may not trigger a Th2-skewed allergic reaction. BSA did not appear to modulate milk immunogenicity to any significant extent.

Bovine κ-casein glycomacropeptide (GMP) is a sialic acid containing glycopeptide having many biological activities. The study described in this research communication was undertaken to determine whether sialic acid rich glycopeptide can be produced from GMP by proteinase treatment. A sample of GMP was hydrolyzed with papain, and the obtained hydrolysate was chromatographed on a column of diethylaminoethyl-Sephacel to obtain a glycopeptide fraction (GPF). This product accounted for average 48.1% dry weight of GMP or 81.1% total recovered sialic acid from GMP. The content of sialic acid (expressed as % dry weight) was 1.7 times higher in GPF (22.6) than in unhydrolyzed GMP (13.4). Major differences in amino acid composition between GPF and GMP were found in the contents (mol%) of: lysine (<1 and 4.5, respectively), serine (20.3 and 10.3, approximately twice higher in GPF), asparagine/aspartic acid and isoleucine. The contents of the last two amino acids were approximately twice lower in GPF. On gel filtration chromatography with Sephacryl S-100, GMP was eluted as a single peak with elution volume similar to that of dimeric β-lactoglobulin (36.6 kDa) whereas GPF was eluted in two peaks both with elution volumes greater than that of α-lactalbumin (14.2 kDa). These peak fractions containing high (fraction I) and low (fraction II) molecular size glycopeptides gave different sialic acid to peptide ratio, which was 1.7 times higher in fraction I than in fraction II. Results of size exclusion HPLC on Superdex-75 were consistent with those of gel filtlation chromatography. On cellulose acetate electrophoresis, the mobility of GPF relative to that of GMP as 1.0 was found to average 1.2, suggesting a higher negative charge density in GPF than in GMP. It was concluded that papain digestion of GMP is an efficient method to produce glycopeptide with high sialic acid content.

This Technical Research Communication evaluated the influence of various cheese manufacture methods on the composition and in vitro antihypertensive activity of sweet whey samples obtained from miniature models for fresh, Chanco and Gouda-style cheese processing using bulk-tank milks throughout a year. Raw milks from morning milking were standardized, pasteurized and used to obtain sweet whey using cheesemaking protocols for each variety on 200 g scale, as well the use of whey dilution at levels of 0, 15, 30 and 45% in Chanco and Gouda-style making. The composition of sweet whey obtained within each cheesemaking variety was similar among different timepoints of the year (P > 0.05), which was attributed to similar composition of milks and the use standardized cheesemaking protocols used for this study. As expected, the use of whey dilution led to sweet whey samples with reduced levels of total solids (P < 0.05), but they exhibited an improvement of the in vitro antihypertensive properties, which may be attributed to the formation of low-molecular weight bioactive peptides due to increased cheese making times. The results of this study suggest that modifying cheese manufacture protocols may have a direct impact on the bioactive properties of sweet whey. Future work will be required to identify and evaluate the feasibility to purify bioactive peptides obtained from sweet whey.

The experiments reported in this research paper aimed to determine the technological properties of indigenous Lactococcus lactis strains isolated from Lait caillé, a spontaneous fermented milk, from the perspective of starter culture development. Fermentations were conducted to determine the acidification patterns. The ropy character, growth in 0.04 g/ml NaCl and citrate metabolism were additionally tested. Furthermore, the rheological properties of samples from selected strains and the impact of cold storage were evaluated. Based on the rate of acidification, the indigenous strains were divided into 2 groups depending on their fermentation time, i.e. 10–13 h (fast acidifier), and up to 72 h (slow acidifier), respectively. The physiological tests suggested that most of these strains produced exopolysaccharides but none could ferment citrate. The flow properties of the samples inoculated by the fast acidifier strains showed a time-dependent shear thinning behaviour, while their viscoelastic properties corresponded structurally to those of weak gels. Cold storage decreased the viscosity and CFU counts for most of the indigenous strains tested. This study is a step towards the definition of starter cultures for African spontaneous fermented milks such as Lait caillé.

This research communication addresses the impact of the addition of Lactobacillus casei and/or carbonation (CO2) on the chemical composition, physicochemical characteristics, probiotic survival, and sensory acceptance of passion-fruit flavored whey dairy beverages (70% milk/30% whey) during storage (30 d/4°C). The addition of Lactobacillus casei and/or carbonation did not impact on the chemical composition, pH values, and acceptance (flavor and overall impression) of the products, but increased the acidity, and decreased the aroma acceptance. The carbonation process did not affect the probiotic survival but decreased the acidity of the products during storage. It can be concluded that it is possible to develop a probiotic passion-fruit flavored carbonated whey dairy beverage with suitable chemical composition, acidity, sensory acceptance (>6 in 9-point hedonic scale) and probiotic viability (>7 log cfu/ml) that could be refrigerated stored for 30 d. This is the first report considering a probiotic non-fermented carbonated whey dairy beverage.

In this Research Reflection we describe a common standpoint on suitable methodology for controlled and observational studies in cow-calf contact systems in dairy production. Different methods to assess behaviour, health and production in cow-calf contact systems are outlined. Knowledge and experience from researchers working in this field supplement scientific literature whenever relevant. Specific methods including study design, early behaviour of cow and calf, social behaviour relevant to cow-calf contact systems, human-animal relationships and aspects related to management (milking, weaning and separation, health) are reviewed, and recommendations formed. We expect that this paper can contribute to a better understanding of the complexity of cow-calf contact systems and help to advance research in this area of dairy production.

This research paper addresses the hypothesis that the application of ultraviolet (UV) light before packaging of pasta-filata cheese has the potential to eliminate or control post-processing contamination whilst maintaining chemical and sensorial quality. The surfaces of kashar cheese were treated at different doses of UV light (0.32–9.63 kJ/m2) in a batch UV cabinet system to determine effects on physicochemical and sensorial quality as well as mould inactivation. Untreated cheese samples were also used for comparison. Kashar cheese was naturally contaminated in a mouldy environment to provide the desired mould numbers before UV treatments. Log reductions of 0.34, 0.69 and 2.49 were achieved in samples treated at doses of 0.32, 0.96 and 1.93 kJ/m2, respectively and the mould count of sample treated at 9.63 kJ/m2 was below the detection limit. We found no significant differences in composition and hardness values between any of the treated or control cheeses. Although some individual colour values increased as the UV doses increased, this change was not observed visually in sensory analysis. Increased light intensity accelerated the lipid oxidation causing a perception of off-flavour. The results of this study show that it is necessary to examine the relationship between the oxidative and sensory interactions while determining the effective doses applied to cheese surface for microbial inactivation.

Identification of the most proteolytic Pseudomonas strains that can produce heat-resistant proteases and contribute to the Ultra High Temperature (UHT) milk destabilization is of great interest. In the present study, among the 146 Pseudomonas isolates that encoded the aprX gene, five isolates with the highest proteolytic activity were selected and identified based on 16S rRNA, rpoD and gyrB gene sequences data. The identification results were confirmed by phylogenetic analysis based on multilocus sequence analysis and identified the representative isolates as P. jessenii (two isolates) and P. gessardii (three isolates). Casein zymography demonstrated the ability of these species to produce heat-resistant enzymes, AprX, with molecular mass of about 48 kDa during storage at 7° C for 72 h. In sterilized milk samples, the residual activity of AprX caused a considerable enhancement in the degree of protein hydrolysis, non-protein nitrogen and non-casein nitrogen contents of the samples during a two-month storage. This enhancement was slightly higher in samples containing enzyme produced by P. jessenii compared to P. gessardii ones, resulting in earlier onset of sterilized milk destabilization. Hence, this study revealed that P. jessenii and P. gessardii can play a considerable role in deterioration of Iranian commercial long-life milk.

In this research paper we describe the technological properties of beneficial lactic acid bacteria (LAB) obtained from a dairy production chain and the development of a fermented milk produced with Lactobacillus casei MRUV6. Fifteen LAB isolates (Lactobacillus sp., Pediococcus sp. and Weissela sp.) presented acidifying abilities (pH ranges from 0.73 to 2.11), were able to produce diacetyl (except by 5 isolates) and exopolysaccharides, and two were proteolytic. L. casei MRUV6 was selected for producing a fermented milk, stored up to 35 d at 4 and 10°C. Counts on MRS agar with added vancomycin (10 mg/l) and MRS agar with added bile salts (1.5% w/v) ranged from 9.7 to 9.9 log CFU/g, independently of the tested conditions, indicating stability and intestinal resistance of L. casei MRUV6, despite some significant differences (P < 0.05). The study demonstrated the technological potential of a potential probiotic candidate strain, L. casei MRUV6, to be used as a starter culture in the dairy industry.

We analyzed the effects of growth of Penicillium commune, one of the most frequent fungal species associated with cheese, on the water diffusion and texture of hard and extra-hard cheeses. A total of 36 hard cheese blocks and 36 extra-hard cheese blocks were manufactured, salted at different levels (0.5, 1.25 and 2% w/w), and assigned to different treatments (control and inoculated). Cheese texture was analyzed using a penetration needle probe at 2 and 5 weeks after ripening. Firmness, defined as the maximum force detected in the penetration probe, was higher in both hard and extra-hard inoculated cheese blocks compared with the control. In addition, the presence of fungal growth on cheese rind increased the total work of penetration (a measure of resistance to probe penetration), but only in extra-hard cheeses, suggesting that the moisture of cheese might be affecting the growth capacities and performance of P. commune. The change in textural properties of cheeses was linked to desiccation of the upper 0.5-cm rind layer mediated by P. commune.

This position paper describes a common stand on methodology of human attitudes and behaviour that is suitable to use in studies regarding cow−calf contact (CCC) in dairy production, in order to create a common knowledge base and foundation for future recommendations of CCC systems. We describe how different quantitative and qualitative methods can be used to study human attitudes to CCC as well as farmer or consumer behaviour. We aim to contribute to a better understanding of the available methods, and hope that this paper can be used as a guideline for future studies in this area.