This research paper aimed to locate protein modifications caused by treatment of milk and determine if the modification locations were consistent. The majority of milk for consumption is homogenised using pressure and heat, and this causes changes in the location of proteins in the milk as well as protein modifications. To investigate these proteomic changes, raw milk was pasteurised (72°C, 15 s), then, to separate the treatment for homogenisation, heated at these different pressures and temperatures: 45°C without no pressure applied, 45°C with 35 MPa, 80°C without pressure applied and 80°C, with 35 MPa. Proteomic analysis was done after separating the milk into three fractions: whey, casein and cream. Protein modifications in each fraction were examined and we found Maillard products as well as oxidation to be of interest. The proteins were also further identified and characterised to compare protein modification sites and differences in proteins present in the cream resulting from homogenisation and/or pasteurisation. This experiment showed that both heat and pressure during homogenisation can cause increases in protein modifications as a result of oxidation or the Maillard reaction. Total cysteine oxidation and total proline oxidation differed between treatments although this was only significantly different for cysteine. It was observed that protein modifications occurred in the same location in the protein sequence rather than in random locations which we highlighted by examining α-S1-casein, lactadherin and β-lactoglobulin.

Functions of mucin, as the major macromolecular component in saliva or gastric fluids, are drawing increasing attention in the context of understanding the oral processing or digestion of dairy foods at the molecular level. This study was designed to investigate the interactions between β-lactoglobulin (BLG)-lactose, mucin-lactose and BLG-lactose-mucin at the molecular level under different temperature and pH conditions using fluorescence spectroscopy in combination with scanning electron microscopes (sem). It is the first study of its kind. There was no lactose-dependent quenching on BLG fluorophore in the range of 0–10 mM lactose concentration. On the contrary, there was a continuous increase in the fluorescence intensity of the BLG protein when the lactose concentration increased, especially at 25°C. BLG-lactose complex became thermally unstable at 37 and 45°C. Moreover, BLG exhibited a pH dependent conformational change and had higher fluorescence intensity at pH 3 than pH 6.8. The fluorescence result was in correspondence with sem images where we observed lactose crystals gathering around and on the BLG molecule, but lactose molecules could not be seen in the presence of mucin. It was anticipated that mucin molecules interacted with BLG-lactose complex via electrostatic attraction and formed an extra protective layer around the BLG molecules to avoid solvent exposure.

In this research communication we compare three different approaches for developing dry matter intake (DMI) prediction models based on milk mid-infrared spectra (MIRS), using data collected from a research herd over five years. In dairy production, knowledge of individual DMI could be important and useful, but DMI can be difficult and expensive to measure on most commercial farms as cows are commonly group-fed. Instead, this parameter is often estimated based on the age, body weight, stage of lactation and body condition score of the cow. Recently, milk MIRS have also been used as a tool to estimate DMI. There are different methods available to create prediction models from large datasets. The main data used were total DMI calculated as a 3-d average, coupled with milk MIRS data available fortnightly. Data on milk yield and lactation stage parameters were also available for each animal. We compared the performance of three prediction approaches: partial least-squares regression, support vector machine regression and random forest regression. The full milk MIRS alone gave low to moderate prediction accuracy (R2 = 0.07–0.40), regardless of prediction modelling approach. Adding more variables to the model improved R2 and decreased the prediction error. Overall, partial least-squares regression proved to be the best method for predicting DMI from milk MIRS data, while MIRS data together with milk yield and concentrate DMI at 3–30 d in milk provided good prediction accuracy (R2 = 0.52–0.65) regardless of the prediction tool used.

This research paper addresses the hypothesis that powdered milk may contain amyloid fibrils. Amyloids are fibrillar aggregates of proteins. Up to this time, research on the presence of amyloids in food products are scarce. To check the hypothesis we performed thioflavin T fluorescence assay, X-ray powder diffraction, atomic force microscopy and fluorescence microscopy imaging. Our preliminary results show that commercially available milks contain fibrils that have features characteristic to amyloids. The obtained results can be interpreted in two opposite ways. The presence of amyloids could be considered as a hazard due to the fact that food products may induce amyloid related diseases. On the other hand, the presence of amyloids in traditionally consumed foodstuffs could serve as proof that fibrils of food proteins do not pose a threat for consumers.

This research communication reports the responses to supplementing dairy cattle with a hydrogenated fat-embedded calcium gluconate feed additive. The role of hindgut health in ruminant performance and wellbeing is an area of growing interest. Various prebiotic compounds have been used to promote lower gut health in various non-ruminant species. Calcium gluconate, a prebiotic compound, has previously been observed to increase milk fat yield when fed to ruminants in a form capable of resisting fermentation in the rumen, though the mechanism(s) behind this response remain unclear. The objective of this study was to compare the responses of lactating cattle to two different supplementation levels of a hydrogenated fat-embedded calcium gluconate (HFCG) product to evaluate a potential linear dose response. Forty-six lactating Holstein dairy cattle were used in a 3 × 3 replicated Latin square design with 28 d periods to evaluate a previously used dose of HFCG (approximately 16 g/d) with both a negative control and a dose of 25 g/d. Supplementation of multiparous animals with 16 g/d HFCG significantly (P < 0.05) increased milk fat yield and content relative to the negative control, and subsequently improved gross feed efficiency (P < 0.05); additionally, the presence of a potential non-linear dose response was observed for these parameters. Responses when supplemented with 25 g/d HFCG did not differ from the negative control. No production responses were observed in primiparous animals. The mode of action of HFCG, in addition to the potential differential response in primiparous animals remains unclear and warrants further investigation.

This research paper assessed textural, microstructural, sensory and colour properties of set-yoghurts produced using milk from two indigenous cattle types, Thamankaduwa White (TW) and Lankan cattle (LC) compared to two generic cattle breeds, Friesian and Jersey. Instrumental texture profile (firmness, adhesiveness, cohesiveness and springiness), colour space (L* a* b*) and scanning electron micrographs of set-yoghurts during 21 d of storage (4 ± 1°C) were evaluated. Sensory quality attributes were evaluated with 40 untrained panellists using a five-point hedonic scale. Set-yoghurts prepared using indigenous cow milk showed higher (P < 0.05) firmness, cohesiveness and apparent viscosity values compared to those prepared using generic cow milk. As revealed by micrographs, set-yoghurts made from TW milk had lesser and smaller void spaces and a dense protein gel network than gels made from LC and the two generic breeds. The gel network made from Friesian milk showed a comparatively larger porous gel structure and thinner protein strands resulting in a weaker gel than other milk gels. The highest lightness (L*) and yellowness (b*) were observed from set-yoghurt produced from Friesian and LC milk, respectively. Set-yoghurts from TW milk had the highest (P < 0.05) sensory scores for all sensory attributes. The lowest sensory acceptance was recorded in set-yoghurt made from Friesian milk. Thus, milk from TW and LC is likely to be suitable in producing set-yoghurts with superior textural, microstructural and sensory properties, compared to milk from Jersey and Friesian. Our results suggest the merits of using indigenous cow milk in producing set-yoghurts and, thereby, prioritizing the preservation of the genetic pool of these indigenous breeds.

We compared the potential of dietary lipid supplements of different fatty acid compositions to affect milk performance when early lactation dairy goats were fed a high-concentrate diet. Thirty Alpine goats at 23 ± 5 d in milk were allocated to 1 of 10 blocks according to parity and milk fat concentration. Within each block, goats were randomly assigned to receive, during a period of 41 d, either CONT) a basal diet with a forage to concentrate ratio of 45:55, used as control, or PALM) the basal diet + 2% of a palmitic acid-enriched fat supplement, or FLAX) the basal diet + 7% of extruded flaxseed. Body weight, dry matter intake and milk yield were not different between treatments. As compared with CONT, goats fed PALM and FLAX had a greater milk fat concentration. Moreover, milk fat yield was numerically (but non-significantly) greater with PALM than with CONT. Milk fat from goats receiving PALM had a greater concentration of 16:0 as compared with CONT and FLAX, whereas a greater concentration of cis-9, cis-12, cis-15 18:3 was observed when goats were fed FLAX as compared with CONT and PALM. Under the conditions of the current experiment, dietary fat supplementation had only minor impacts on the yield of major milk constituents, with the exception of a modest increase in fat yield when goats were fed PALM. The impact of a greater concentration of 16:0 in milk fat of goats receiving this feed ingredient on the nutritive value of dairy products remains to be determined.

CXCL8 (also known as IL-8) is a member of the CXC subfamily of chemokines that binds two of the seven transmembrane G-protein-coupled receptors (GPCRs), CXCR1 and CXCR2, to mediate and regulate leucocyte accumulation and activation at sites of inflammation. They are known to play a critical role in both disease susceptibility and infection outcome. The aim of this study was to investigate the entire sequences of CXCL8 and CXCR2 genes in thirty-one Simmental sires to evaluate the effects of genomic variants on the indexes of the bulls for milk, fat and protein yields, and for somatic cell score (SCS). Five new single nucleotide polymorphisms (SNPs) were found in CXCR2 gene. The analysis of association indicated that one SNP in CXCL8 and two in CXCR2 influenced the considered traits. To evaluate the existence of functional haplotypic effects, combinations among the three genomic variants (SNP 1 in CXCL8, SNP 6 and SNP 7 in CXCR2) were investigated. Four different haplotypic alleles were identified in the experimental population, one of which at a high frequency (61%). Bulls with Hap 4 (G-C-G at SNP 1, SNP 6, and SNP 7 respectively) had more favourable indexes for SCS (P < 0.05). These results suggest that the SNPs in CXCL8 and CXCR2 may be potential genetic markers to improve udder health in the Simmental breed.

This study evaluated the impact of three distinct diets; perennial ryegrass (GRS), perennial ryegrass/white clover (CLV) and total mixed ration (TMR), on the sensory properties and volatile profile of whole milk powder (WMP). The samples were evaluated using a hedonic sensory acceptance test (n = 99 consumers) and by optimised descriptive profiling (ODP) using trained assessors (n = 33). Volatile profiling was achieved by gas chromatography mass spectrometry using three different extraction techniques; headspace solid phase micro-extraction, thermal desorption and high capacity sorptive extraction. Significant differences were evident in both sensory perception and the volatile profiles of the WMP based on the diet, with WMP from GRS and CLV more similar than WMP from TMR. Consumers scored WMP from CLV diets highest for overall acceptability, flavour and quality, and WMP from TMR diets highest for cooked flavour and aftertaste. ODP analysis found that WMP from TMR diets had greater caramelised flavour, sweet aroma and sweet taste, and that WMP from GRS diets had greater cooked aroma and cooked flavour, with WMP derived from CLV diets having greater scores for liking of colour and creamy aroma. Sixty four VOCs were identified, twenty six were found to vary significantly based on diet and seventeen of these were derived from fatty acids; lactones, alcohols, aldehydes, ketones and esters. The abundance of δ-decalactone and δ-dodecalactone was very high in WMP derived from CLV and GRS diets as was γ-dodecalactone derived from a TMR diet. These lactones appeared to influence sweet, creamy, and caramelised attributes in the resultant WMP samples. The differences in these VOC derived from lipids due to diet are probably further exacerbated by the thermal treatments used in WMP manufacture.

The experiment reported in this research paper aimed to evaluate the effects of high-starch or starch and oil-supplemented diets on rumen and faecal bacteria, and explore links between the structure of bacterial communities and milk fatty acid (FA) profiles. We used four Holstein dairy cows in a 4 × 4 Latin square design. Cows were fed a diet rich in cereals (high-starch diet with 23% starch content on dry matter (DM) basis), a diet supplemented with saturated FA from Ca salts of palm oil + 18% DM starch, a diet with high content of monounsaturated FA (from extruded rapeseeds) + 18% DM starch or a diet rich in polyunsaturated FA (from extruded sunflower seeds) + 17% DM starch. At the end of each experimental period, cows were sampled for rumen and faecal contents, which were used for DNA extraction and amplicon sequencing. Partial least squares (PLS) regression analysis highlighted diet-related changes in both rumen and faecal bacterial structures. Sparse PLS discriminant analysis was further employed to identify biologically relevant operational taxonomical units (OTUs) driving these differences. Our results show that Butyrivibrio discriminated the high-starch diet and linked positively with higher concentrations of milk odd- and branched-chain FA. YS2-related OTUs were key taxa distinguishing diets supplemented with Ca salts of palm oil or sunflower seeds and correlated positively with linoleic acid in milk. Similarly, diets modulated faecal bacterial composition. However, correlations between changes in faecal and rumen bacteria were poor. With this work, we demonstrated that high-starch or lipid-supplemented diets affect rumen and faecal bacterial community structure, and these changes could have a knock-on effect on milk FA profiles.

In this research communication we describe a straightforward triplex-PCR protocol able to differentiate the origin of milk from three closely related species (goat, sheep and cow) in Halloumi, a cheese with Protected Designation of Origin (PDO), and yogurts. Halloumi must contain at least 51% sheep or goat milk, therefore, the fraudulent adulteration of this cheese with excess of cow milk must be routinely tested. The assay employs one universal forward primer and three species-specific reverse primers giving rise to 287 bp (cow), 313 bp (goat), and 336 bp (sheep) amplicons, under the same amplification conditions. This protocol, when used to test a small number of Cyprus commercial products, correctly detected mislabeling in Halloumi (2 out of 6 samples were adulterated) and yogurt brands (1 out of 4 was adulterated). The suggested protocol is a reliable tool for identifying the origin of milk in Halloumi cheeses and yogurts and can be used in any laboratory equipped with a thermocycler and an agarose gel electrophoresis apparatus.

This research paper addresses the hypothesis that the optimum doses of a continuous type of ultraviolet (UV) light applied to the surface of lor (whey) cheese needs to be identified to maximize mould inactivation and shelf life while minimizing quality deterioration. Therefore, the mould inactivation, protein and lipid oxidation products, sensory and colour attributes of lor cheese subjected to different doses of UV light (1.617, 4.018, and 36.832 kJ/m2) in a continuous type of UV system were evaluated. UV treated samples presented mould counts lower than those of untreated ones. UV treatment at more than 4.018 kJ/m2 allowed around 0.7–2.7 log reductions on mould growth during storage. The increase in UV light dose caused significant increases in primary and secondary lipid oxidation products. In particular, the highest doses applied to the surface of cheese samples had the highest values of protein carbonyls, as well as lipid oxidation products. Strong positive correlations were recorded between lipid and protein oxidation markers. Exposure to the highest doses of UV light increased foreign flavour perception, probably due to the oxidative reactions. The results indicated that the application of UV light to the lor cheese surface allowed delaying mould growth during storage but extreme doses could induce lipid and protein oxidation reactions, leading to quality deterioration.

Bakery by-products (BP), rich in fats and sugars, are unconventional feed sources for cows whose effects on milk fat composition have not yet been evaluated. This research paper aimed to assess the effects of dietary BP inclusion rate and feeding period on the milk fatty acid composition. Twenty-four Simmental cows were fed a diet without BP (CON) for 1 week. Then they either continued with the CON diet or switched to one of the BP diets (with 15% or 30% BP in diet dry matter) for 3 weeks. Milk samples were taken before diet change and three times during BP feeding and analysed for fatty acid composition. Data showed that increasing BP content in the diet increased total fatty acid intake, especially of 18 : 1 n9. In the milk fat, the percentages of total monounsaturated fatty acids especially of the 18 : 1 origin linearly increased with increasing dietary BP level. The percentage of fatty acids de novo synthesized in the mammary gland (the sum of 4 : 0–14 : 0) remained similar among diets (32-34% of total fatty acids). The 16 : 0 percentage dropped from 32.5 to 29.6% and from 33.6 to 28.3% for 15% and 30% BP, respectively. Only 30% BP elevated the percentage of conjugated linoleic acids (CLA: by 59%) compared with CON throughout the 3 weeks. Proportions of 18 : 2 n6 and 18 : 3 n3 and the n6:n3 ratio were unaffected by BP and feeding time. BP feeding improved all those estimated health indices of the milk fat that are suggested to be related to coronary health. In summary, the inclusion of BP in dairy rations beneficially shifted the milk fatty acid profile to more 18 : 1 fatty acids at the expense of 16 : 0. At a 30% inclusion rate, BP feeding showed an additional benefit of increased CLA content in milk fat.

This research communication reports concentrations of two sialic acids (SA), N-acetylneuraminic (Neu5Ac) and N-glycolylneuraminic (Neu5Gc), in fresh milk from different cow breeds throughout lactation. According to published studies, the two SA types found in animal-derived products have diverse and conflicting effects on human health, but SA content is not routinely analysed in individual milk cows samples. We measured the content of Neu5Ac and Neu5Gc in milk from Holstein Friesian (HO), Simmental (SM), Simmental × Holstein crossbred (SM×HO), and Podolica (POD) cows at 60 and 120 d following calving. HO, SM and SM×HO were reared in an intensive production while POD were raised in an extensive system. Results showed that total Neu5Ac was overall thirty times more abundant than Neu5Gc, and their concentrations were higher at 120 d than at 60 d (P < 0.001). Neu5Gc values were greater in HO, SM, and SM × HO than in POD (P < 0.001), while HO had a higher Neu5Ac value than the other three breeds (P < 0.001). These findings shed light on the differences in SA content among cow breeds and lay the groundwork for future research to select animals that produce milk with desirable characteristics for human health.

This study was designed to show the changes in glycolmacropeptides (GMPs) of whey protein solution (WPS) due to different pretreatments before and after ultrafiltration (UF). The combined form of two variants (A&B) of GMPs is a helpful compound for nutritional management of phenylketonuria and ulcerative-colitis diseases and has low content of phenylalanine (Phe). WPS with 10% concentration was prepared, acidified (adjusted to pH = 3.0), and passed through a PES (polyethersulfone) membrane in the 1st-stage of ultrafiltration (UF-1). Then the resulting permeate was neutralized and went through the 2nd-stage of ultrafiltration (UF-2) under similar conditions. Four experiments of TRT-CON, CON-TRT, TRT-TRT, and CON-CON were used with different pretreatments, where TRT was a mixing-treatment of 30 min at 150 RPM applied either after acidification of WPS or after neutralization of first permeate and before UF-2 process. While the concentration and purity of the combined GMPs in UF-2 retentate in TRT-TRT respectively were >95.6 and 99.5%, its Phe became <10 ppm among the experiments. Highly glycolyzed polymers of GMPs (MW = 45–50 kDa) were formed in the TRT-TRT experiment and went through the pore sizes of PES membrane of UF-1 easily because of their flexible structure. However, they remained in the UF-2 retentate, due to to the formation of bulky polymers. The TRT-TRT experiment had the highest reversible and irreversible resistances for passing through the UF-1 and remaining on the UF-2 membranes, and its fouling index was significantly less than other experiments.

This study aimed to evaluate the effect of an extended voluntary waiting period (VWP) on SCC, SCC elevations and clinical mastitis incidence during the complete lactation and the first 6 weeks of the next lactation. Holstein-Friesian dairy cows (N = 154) were blocked for parity, expected milk yield, calving season and breeding value for persistency and were randomly distributed across 3 VWP (50, 125, or 200 d: VWP-50, VWP-125, VWP-200). Cows were monitored from calving until 6 weeks into the next lactation, or until culling. An elevation of SCC in milk was defined as SCC in milk ≥200 000 cells/ml after two previous weeks with SCC < 200 000 cells/ml. Over the complete lactation, extending the VWP did not affect SCC elevations and the occurrence of clinical mastitis per lactation or per cow per year. There was no clear effect of VWP length on SCC in the complete lactation, except that multiparous cows in VWP-125 had a higher SCC compared with multiparous cows in VWP-50. Dry-off antibiotic usage per cow per year was lower in VWP-200 compared with VWP-50 for multiparous cows. In the first 6 weeks of the next lactation, cows in VWP-200 had a higher SCC compared with cows in VWP-50, with no effect of VWP on the number of elevations of SCC or the occurrence of clinical mastitis. Extending the VWP may therefore be used to reduce the frequency of transition periods and the associated use of dry-cow antibiotics, with limited impact on udder health, and a similar occurrence of SCC elevations and clinical mastitis per year.

The objective of this retrospective longitudinal study was to evaluate the relationship between dry period length and the production of milk, fat, protein, lactose and total milk solids in the subsequent lactation of Holstein dairy cows under tropical climate. After handling and cleaning of the data provided by the Holstein Cattle Breeders Association of Minas Gerais, data from 32 867 complete lactations of 19 535 Holstein animals that calved between 1993 and 2017 in 122 dairy herds located in Minas Gerais state (Brazil) were analysed. In addition to dry period length, calving age, lactation length, milking frequency, parity, calf status at birth, herd, year, and season of calving were included in the analysis as covariables to account for additional sources of variation. The machine learning algorithms gradient boosting machine, extreme gradient boosting machine, random forest and artificial neural network were used to train models using cross validation. The best model was selected based on four error metrics and used to evaluate the variable importance, the interaction strength between dry period length and the other variables, and to generate partial dependency plots. Random forest was the best model for all production outcomes evaluated. Dry period length was the third most important variable in predicting milk production and its components. No strong interactions were observed between the dry period and the other evaluated variables. The highest milk and lactose productions were observed with a 50-d long dry period, while fat, protein, and total milk solids were the highest with dry period lengths of 38, 38, and 44 d, respectively. Overall, dry period length is associated with the production of milk and its components in the subsequent lactation of Holstein cows under tropical climatic conditions, but the optimum length depends on the production outcome.

Greek yogurt is one of the fastest growing products in the dairy industry. It is also known as strained yogurt, which is obtained after draining the whey. As a result of the draining process, Greek yogurt has higher total solids and lower lactose than regular yogurt. Since it is a concentrated yogurt, its sensory characteristics are different from regular yogurt. However, there is little information about factors influencing the quality of Greek yogurt and sensory evaluation techniques applied to Greek yogurt. This review aims to describe the effects of ingredients, starter cultures, processing techniques and other parameters on quality characteristics and sensory properties of Greek yogurt. In addition, advantages and limitations of novel sensory evaluation techniques applied to Greek yogurt products are discussed. In particular, we take a look at advanced techniques such as the electronic nose and electronic tongue and the benefits of these techniques with regard to Greek yogurt. This review should help the Greek yogurt industry to improve its current products and develop innovative products based on appropriate food evaluation techniques.

The aim of this research communication was to examine and report the current situation of dairy farms in the province of Lugo (Galicia, Spain) regarding facility problems. We assessed the facilities of 168 free-stall dairy farms, housing in total 9228 Holstein cows in milk. Housing factors related to the resting area, circulation area, feeding area, ventilation area and milking area, as well as animal-handling features, were evaluated. Distance measurements were performed using a laser metre or a roll metric tape. A survey was conducted to gather information about cleaning and preventive protocols. Our results showed that most farms do not comply with the objective for cubicle measurements, width of the crossovers, type of flooring and presence of a quarantine pen, which may have a negative impact on the health and productivity of the animals. Therefore, to maximise the profits of the farm, the recommendations given in this study may be useful as a guide when building a new farm or remodelling the existing ones.

Bovine κ-casein glycomacropeptide (GMP) is a sialic acid containing glycopeptide, which is considered as a health promoting compound found in cheese whey. The study described in this research communication was undertaken to determine whether GMP with undetectable level of contaminating protein or phenylalanine can be isolated from bovine whey fraction using batch anion exchange technique with chitin as an adsorbent. A soluble whey fraction (SWF) prepared from 1 g whey protein isolate (WPI) was mixed with a slurry of 1 g chitin, and the mixture was incubated at pH 3.0. After incubation, the mixture was filtered, and the residue obtained (containing chitin-GMP complex) was washed with water and eluted stepwise with 0.5 M NaCl and 2.0 M NaCl. Most of GMP (corresponding to 75.8% of total sialic acid recovered) was eluted with 0.5 M NaCl. The recovered GMP accounted for 5.4% dry weight of WPI (or 18.9% dry weight of SWF). Amino acid analysis showed that there was no detectable level of contaminating amino acids including phenylalanine, histidine, arginine and tyrosine in the GMP fraction. It was concluded that the batch anion exchange method with chitin developed in this study can be used for the isolation of high purity GMP from bovine SWF.