Journal of Dairy Research

Changes in chemical composition, proteolysis, lipolysis, texture, melting and sensory properties of low-fat Kashar cheese made with three different fat replacers (Simplesse® D-100, Avicel Plus® CM 2159 or β-glucan) were investigated throughout ripening. The low-fat cheeses made with fat replacers were compared with full- and low-fat counterparts as controls. Reduction of fat caused increases in moisture and protein contents and decreases in moisture-in-non fat substance and yield values in low-fat cheeses. The use of fat replacers in the manufacture of low-fat Kashar cheese increased water binding capacity and improved overall quality of the cheeses. Use of fat replacer in low-fat cheese making has enhanced cheese proteolysis. All samples underwent lipolysis during ripening and low-fat cheeses with fat replacers had higher level of total free fatty acid than full- or low-fat control cheeses. Texture attributes and meltability significantly increased with addition of fat replacers. Sensory scores showed that the full-fat cheese was awarded best in all stages of ripening and low-fat variant of Kashar cheeses have inferior quality. However, fat replacers except β-glucan improved the appearance, texture and flavour attributes of low-fat cheeses. When the fat replacers are compared, the low-fat cheese with Avicel Plus® CM 2159 was highly acceptable and had sensory attributes closest to full-fat Kashar cheese.

The principal chemical components of milk from the Arabian camel (Camelus dromedarius) were monitored in Jordan over one year. The analyses included total solids, fat, protein, vitamins, minerals and organic acids. Large seasonal variations in total solids and fat were apparent, with maxima in mid-winter of 139 and 39·0 g/l, respectively, and minima in August of 102 and 25·0 g/l. These differences may be sufficient to alter the sensory properties of the milk, and the fat: casein ratio may need standardisation for cheesemaking. The mean values of trace elements like zinc (5·8 mg/l), iron (4·4 mg/l) and manganese (0·05 mg/l) in Jordanian camel milk could provide valuable additions to the diet of urban populations, as could the mean concentration of vitamin C (33 mg/l). The levels of organic acids were generally higher than in bovine milk and, as with all the constituents of the milk, there were discernible patterns linking concentration and season of the year.

The dynamics of non-esterified fatty acid (NEFA) metabolism in lactating dairy cows requires quantification if we are to understand how dietary treatments and disease influence changes in body condition (adipose reserves) and the production of milk fat. We present here a novel compartmental model that employs the pattern of plasma glucose concentrations to predict the dynamic changes that occur in plasma NEFA concentrations during an intravenous glucose tolerance test (IVGTT) in lactating dairy cows. The model was developed using data obtained from ten early-lactation, Holstein-Friesian cows given a standard IVGTT. The model described all of the major features of the NEFA response to an IVGTT; it was consistent with physiological processes and provided a number of parameters that can be used to quantify NEFA production and utilization. For all of the individual cows, all model parameters were well identified and usually had CV<10% of their estimated values. In the model, elevated plasma glucose concentrations cause an increase in the level of glucose in a remote compartment, which in turn suppresses the rate of entry of NEFA to the plasma compartment. The means (±sd) for the five adjustable parameters of the model were: rate of entry of NEFA to the plasma pool (SFFA) 183±71 [μmol l−1 min−1], rate of removal (oxidation, sequestration in adipose tissue and uptake by the mammary gland for milk production) of NEFA from the plasma pool (KFFA) 0·140±0·047 [min−1], a threshold parameter (gs) representing a plasma glucose concentration above which elevated levels of plasma glucose result in entry of glucose into a ‘remote’ or inaccessible glucose compartment, 3·30±0·52 [mmol/l], a rate constant (K) describing the movement of plasma glucose (above gs) into a remote compartment 0·063±0·033 [min−1] and a parameter Φ which is a Michaelis Menten type affinity constant which modulates the extent to which remote glucose inhibits the provision of NEFA to the plasma pool, 0·812±0·276 [mmol/l]. It is concluded that the model is suitable to describe NEFA kinetics in lactating dairy cows and it may have application in other species.

This study was conducted to determine the effect that relocation to a new free stall barn had on locomotion and cleanliness of two breeds of dairy cows. The original facility before relocation had cows housed in an 8-row free stall barn. Cows were allocated in a new 4-row free stall facility: cows of two breeds (n=22 Holsteins and 22 Jerseys) were intermixed in the northwest section. Locomotion (scale 1–5) and cleanliness were scored (scale 1–4). Holsteins and Jerseys had no difference in locomotion score throughout 12 weeks following relocation. A lactation number by date interaction showed cows in third and greater lactations had significantly higher locomotion scores (more lameness) by day 86. Locomotion scores increased across breeds during the 86-d observation period, suggesting cows became lamer. Jerseys had cleaner lower legs than Holsteins (2·9±0·1 v. 3·5±0·1, respectively). Lactation number affected lower leg cleanliness, with scores decreasing as lactation number increased (3·4 v. 3·3 v. 2·9±0·10 for first, second and third and greater lactations, respectively; P<0·01). All cows were cleaner (lower scores) after relocation, suggesting that the new facility improved hygiene.

Human milk samples from three healthy donors were investigated in order to evaluate the antibacterial activity during lactation against Escherichia coli ATCC 25922 and Listeria monocytogenes. The concentration of the main human-milk antimicrobial proteins (lactoferrin (LF), lysozyme (LZ) and secretory immunoglobulin A (sIgA)) was determined by ELISA. Results showed that human milk exhibited antibacterial activity against List. monocytogenes, although it was weakly active against Esch. coli ATCC 25922. The observed antilisterial activity was positively correlated with LZ concentration. In addition, the effect of gastrointestinal proteases, at different pH conditions, that prevail in the stomach of infants (pH 2·0–6·5), on antilisterial activity and protein degradation was evaluated. Hydrolysis with pepsin at pH 4·0–6·5, followed by treatment with pancreatic enzymes, resulted in a decreased hydrolysis of LZ, LF and sIgA and an enhanced antibacterial activity against List. monocytogenes. It is suggested that partial degradation of certain milk proteins at the gastrointestinal level may produce peptides that could act synergistically with the remnant intact proteins.

Lactococcus lactis strain AMP2I expresses OppA(D471R), a mutant oligopeptide binding OppA protein in which the aspartyl residue at position 471 was replaced by arginine. As a consequence of a different peptide transport in this strain, experimental Hispánico cheese made with Lc. lactis AMP2I had a higher content of total free amino acids than control cheese made with Lc. lactis AMP1I, an isogenic strain expressing wild-type OppA (Picon et al. 2005, 2007). In this work higher levels of diketones, hydroxy-ketones and, to a lesser extent, branched chain aldehydes were recorded for experimental cheese compared with control cheese. These differences levelled off as ripening proceeded. Strong correlations support the hypothesis that the increased levels of these volatile compounds in cheese made with Lc. lactis AMP2I are linked to higher concentrations of free amino acids threonine, valine and leucine.

In order to have a deeper insight into the retinol isomerization phenomenon, in this work different milk samples have been analysed for their content of trans retinol and its cis-isomers, by means of reliable HPLC techniques. Levels of the different isomers and the degree of retinol isomerization (13-cis/all-trans ratio, %) have been monitored during milk storage at different temperatures and after addition of specific microorganisms. In raw milk stored at 4°C for 96 h the degree of retinol isomerization shifted from 1·1 to 2·3%, while in raw milk stored at 22°C for 24 h it increased from 1·1 to 12·7%. Among microorganisms tested in pasteurized milk, the most active in causing an increment in the 13-cis/all-trans ratio (%), from 3·4 to 33·4% in 8 h, was Streptococcus thermophilus. The results obtained demonstrated a relationship between microbial evolution and retinol isomerization. Therefore, the determination of retinol isomers is of importance not only for a more precise evaluation of vitamin A activity but also for the evaluation of the microbiological quality of milk.

In this study, the properties of casein particles reformed from alkaline disrupted casein micelles were studied. For this purpose, micelles were disrupted completely by increasing milk pH to 10·0, and subsequently reformed by decreasing milk pH to 6·6. Reformed casein particles were smaller than native micelles and had a slightly lower zeta-potential. Levels of ionic and serum calcium, as well as rennet coagulation time did not differ between milk containing native micelles or reformed casein particles. Ethanol stability and heat stability, >pH 7·0, were lower for reformed casein particles than native micelles. Differences in heat stability, ethanol stability and zeta-potential can be explained in terms of the influence of increased concentrations of sodium and chloride ions in milk containing reformed casein particles. Hence, these results indicate that, if performed in a controlled manner, casein particles with properties closely similar to those of native micelles can be reformed from alkaline disrupted casein micelles.

We evaluated the effect of grazing time of day on goat milk chemical composition, renneting properties and milk fatty acid profile in a Mediterranean grazing system. Sixteen lactating Girgentana goats were divided into two experimental groups and housed in individual pens, where they received 500 g/d of barley grain. For 5 weeks the two groups were left to graze in two fenced plots on a ryegrass sward as follows: morning group (AM), from 9·00 to 13·00; afternoon group (PM), from 12·00 to 16·00. In selected herbage, water-soluble carbohydrates (WSC) increased in the afternoon (204 v. 174 g/kg dry matter, DM; P=0·01), whereas crude protein (CP) and linolenic acid decreased (respectively, 16·7 v. 19·8% DM; P<0·01 and 26·8 v. 30·4 g/kg DM; P<0·01). Pasture dry matter intake (DMI) was significantly higher in the afternoon (0·82 v. 0·75 kg/d; P=0·026). Fat corrected milk production (FCM), milk fat and lactose content were not affected by treatment, whereas protein and titrable acidity (°SH) increased in the PM group (respectively 3·56 v. 3·42%; P=0·01; 3·55 v. 3·22°SH/50 ml; P=0·01). In contrast, milk urea content was significantly higher in the AM group (381 v. 358 mg/l; P=0·037). The results seem to indicate that an improvement in ruminal efficiency might be obtained by shifting grazing time from morning to afternoon, as a consequence of a more balanced ratio between nitrogenous compounds and sugars. Indeed, the higher linolenic acid and the lower conjugated linoleic acid (CLA) (respectively 1·02 v. 0·90, P=0·037; 0·71 v. 0·81% of total fatty acids, P=0·022) in the milk of goats grazing in the afternoon seem to indicate a reduced biohydrogenation activity in the PM group.

The roles of the pro-adipogenic ligands of the transcription factor Peroxisome Proliferator Activated Receptor gamma (PPARG) in regulating innate immune responses in bovine mammary epithelial cells (bMEC) were investigated using quantitative real-time PCR. The analyses revealed that 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) enhanced the expression of Interleukin 8 (IL-8) and Chemokine (C-X-C motif) ligand 6 (CXCL6) in these cells in a dose-dependent manner. 15d-PGJ2 also induced the expression of transcripts encoding proteins involved in oxidative stress, including Ferritin heavy chain and Superoxide dismutase 1, as well as substantial microfilament reorganization. In contrast, synthetic PPARG agonists displayed a different and much smaller range of effects on the cells, causing down-regulation of Interleukin 1-beta, Interleukin 6 and IL-8 and increased expression of Chemokine (C-C motif) ligand 2 (CCL2) and Tumour necrosis factor alpha (TNFα). In an independent analysis, the cells were pre-incubated with PPARG agonists followed by lipopolysaccharide stimulation. This study revealed that troglitazone increased the responsiveness of the cells to lipopolysaccharide resulting in up-regulation of Interleukin 1-beta, TNFα, IL-8, CCL2 and CXCL6 while 15d-PGJ2 caused down-regulation of TNFα, CCL2 and CXCL6. These findings are relevant to understanding the anti-inflammatory potential of the PPARG ligands and underline different mechanisms of action of 15d-PGJ2 and troglitazone in bMEC. Furthermore, the present results demonstrate that the generation of pro-inflammatory mediators can be modulated by currently available therapeutic agents and may therefore be of value in the treatment of mastitis in ruminants.

We examined the heat stability, somatic cell count (SCC), pH, fat, protein and lactose content of milk from goats during the oestrous period, in order to investigate evidence of possible oestrus effects on milk physical and chemical properties. Goats free from mammary infections were ranked on average SCC from three tests so that they could be stratified randomly in pairs to synchronized oestrus or left as unsynchronized non-oestrus controls. The synchronisation consisted of insertion of an intravaginal progesterone-releasing device for 17 d, and introduction of the bucks the day of the device removal (D0). The repeated measurements analysis of variance model included the fixed effects of the experimental group (oestrus or control) and day and the corresponding interaction and also the random effect of doe. Reduced milk-heat stability, increased SCC, increased protein content and reduced pH were found in the milk samples of the oestrus group on D1, 2 and 3. The fat and lactose content of the milk was not affected by oestrus. These data indicate that the milk of goats during the mating period has reduced heat stability and, therefore, that dilution into bulk tanks should be recommended to avoid clotting when milk is intended for high thermal treatment.

The effects of ultra-high pressure homogenization (UHPH) on skim milk yogurt making properties were investigated. UHPH-treated milk was compared with conventionally homogenised (15 MPa) heat-treated skim milk (90°C for 90 s), and to skim milk treated under the same thermal conditions but fortified with 3% skim milk powder. Results of the present study showed that UHPH is capable of reducing skim milk particle size which leads to the formation of finer dispersions than those obtained by conventional homogenisation combined with heat treatment. In addition, results involving coagulation properties and yogurt characteristics reflected that, when increasing UHPH pressure conditions some parameters such as density of the gel, aggregation rate and water retention are improved.

The somatic cell count (SCC) in milk is associated with increasing proteolytic degradation of caseins and it has been suggested that enzymes derived from somatic cells contribute to a lower yield and poorer quality of cheese. It is essential to increase the knowledge on naturally occurring milk proteinase activities to better understand how to improve the technological quality of milk. The aim of this work was to identify peptides actually present in milk as a result of proteolysis at different levels of SCC and to assign these peptides to potential responsible proteases where possible. Peptide fractions were prepared from acid whey by ultrafiltration at a molecular cut-off value of 10 000 Da. The peptides were separated using capillary reversed phase high performance liquid chromatography (RP-HPLC) and identified by matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometry (MALDI-TOF MS/MS). Peptides identified ranged in mass from 1023 to 2000 Da, and originated from αS1-, αS2- or β-casein. Possible responsible proteases that could be suggested when examining the peptide cleavage sites included plasmin, cathepsin B, D and leukocyte elastase. The results indicated that plasmin was primarily responsible for the observed proteolysis in milk at low cell count, whereas the cathepsins and elastase became implicated at elevated cell count. Specificity and activity of cathepsins and elastase has earlier mainly been studied in model systems, whereas less is known about their activities in milk itself. This is also the first indication of involvement of elastase in milk proteolysis through the unequivocal determination of cleavage sites.

During early lactation, many dairy cows develop fatty liver, which is associated with decreased health and reproductive performance. Currently, fatty liver can be detected reliably only by using liver biopsy followed by chemical or histological analysis, which is not practical in most on-farm situations. We tested whether digital analyses of hepatic ultrasonograms can be used to detect non-invasively fatty liver and estimate liver triacylglycerol content. A total of 49 liver biopsies and ultrasonograms were taken from 29 dairy cows within 2 weeks postpartum. The usefulness of 17 first- or second-order parameters from digital analysis of B-mode ultrasonograms were evaluated by discriminant, correlation, and regression analyses. A group of linear combinations of the 17 parameters correctly classified 40 of 49 samples into normal liver as well as mild, moderate and severe fatty liver when cut-off values were 1%, 5% and 10% and correctly classified 45 of 49 samples when cut-off values were 5% and 10% triacylglycerol of wet weight. A linear combination of 16 image parameters estimated triacylglycerol concentrations of 38 of the 39 liver samples below the cut-off value of 10% within 2·5% of liver wet weight, and a linear combination of 3 parameters estimated triacylglycerol concentrations of the 10 liver samples above the cut-off value of 10% within 2% of liver wet weight. Therefore, ultrasound imaging followed by digital analysis of sonograms has potential to non-invasively detect fatty liver and estimate liver triacylglycerol content.

Twelve spring-calving and twelve winter-calving cows were managed for extended lactation cycles of 18-months duration, with the former group then completing a second extended lactation. Half of the cows were fed according to standard management practice for the herd; the other half received supplementary concentrate from week 9 of lactation onwards. Commencing at the same time, half of the udder of each cow was subjected to increased milking frequency (thrice daily rather than twice daily). Lactation persistency (and hence total milk yield) was significantly increased by frequent milking. Winter calving cows and supplemented cows also exhibited better persistency, but this was only evident up until the point of re-breeding, at around lactation week 33. Milk composition was measured in the spring-calving cows in both their first and second extended lactations. Composition altered during the course of the lactation, protein and fat percentages increasing and lactose percentage decreasing, irrespective of treatment. The quality of the milk for processing into cheese, fermented products, heat-treated products and cream liqueurs was assessed by calculation of casein number (casein protein as a proportion of total protein). Processing quality declined across the course of lactation in those groups that showed poor persistency but not in those that maintained a persistent lactation. Milk hygienic quality (somatic cell counts) showed parallel changes. Body condition score increased during the course of lactation but was not affected by supplementation; none of the cows became excessively fat. All cows remained healthy throughout the extended lactations and the majority (33/36) re-bred successfully. By demonstrating that lactation persistency is plastic and can be improved by simple management interventions, the results lend support to the economic arguments in favour of extended lactation cycles. The likely welfare benefits of extended lactation are also discussed.

Milk culture results at approximately 6 d post calving were assessed in a 2-year retrospective single-cohort study in 178 Norwegian herds. A combined teat dipping and selective antibiotic therapy trial was performed in these herds where cows with composite milk somatic cell count (CMSCC) >100 000 cells/ml before drying-off (geometric mean of the last three CMSCC test-days) and isolation of Staphylococcus aureus or Streptococcus dysgalactiae were selected for either short-acting lactation antibiotic treatment or long-acting dry cow antibiotic treatment. Milk culture results at approximately 6 d post-calving were available from 437 treated cows and 3061 non-treated cows before drying-off and separate multivariable logistic regression models were ran for these two groups. Risk factors associated with isolation of Staph. aureus 6 d post calving for non-treated cows were CMSCC >400 000 cells/ml before drying-off v. <400 000 cells/ml (Odds ratio (OR)=2·4) and clinical mastitis (CM) in the previous lactation v. non-treated (OR=1·5). Risk factors associated with Staph. aureus 6 d post calving for treated cows was a CMSCC >200 000 cells/ml before drying-off v. <200 000 cells/ml (OR=2·3) and CM in the previous lactation versus non-treated (OR=1·7). For non-treated cows it was 1·7-times more likely to isolate Str. dysgalactiae 6 d post-calving if the CMSCC was >50 000 cells/ml compared with <50 000 cells/ml. For treated cows it was 3·7–5·8-times more likely to isolate Str. dysgalactiae 6 d post calving if given short-acting lactation formula at quarter level compared with long-acting dry cow formula used at cow level. Regular use of iodine post-milking teat disinfection (PMTD) did not influence the isolation of Staph. aureus 6 d post calving, but it was less likely to isolate Str. dysgalactiae 6 d post calving if iodine PMTD was used regularly rather than irregularly. The external teat sealant had no effect on either of the two bacteria.

This study indicates that the CMSCC limit for sampling cows before drying-off can be reduced to 50 000 cells/ml in herds with a Str. dysgalactiae problem. Iodine PMTD should also be recommended in these herds. Cows with a CMSCC >400 000 cells/ml prior to drying-off should receive long-acting dry cow formula irrespective of the milk culture result.

This study describes a method for species-specific detection of animal DNA from different species (cattle, sheep, goat, water buffalo) in milk and dairy products. A primer set was designed in conserved region on the basis of the alignment of the sequence codifying the genomic κ-casein gene in order to amplify all four species with a single primer pair. Polymorphisms were detected via minisequencing with extension primers designed in conserved sequences for haplotype determination that allow unambiguous assignment to each species. The method was successfully applied to the detection of raw and pasteurized milk from the four different species considered as well as to cheese products from the retail trade. Estimation of the limit of detection was carried out using a progression of dilutions of genomic DNA as well as DNA isolated from milk of a known number of somatic cells from different species in order to be able to achieve detection rates as low as 0·1% bovine milk mixed with buffalo milk.

A model to calculate the economic losses of mastitis on an average Dutch dairy farm was developed and used as base for a tool for farmers and advisors to calculate farm-specific economic losses of mastitis. The economic losses of a clinical case in a default situation were calculated as €210, varying from €164 to €235 depending on the month of lactation. The total economic losses of mastitis (subclinical and clinical) per cow present in a default situation varied between €65 and €182/cow per year depending on the bulk tank somatic cell count. The tool was used to measure perception of the total economic losses of mastitis on the farm and the farmers' assessment of the cost factors of mastitis on 78 dairy farms, of which 64 were used for further analyses. Most farmers (72%) expected their economic losses to be lower than those revealed by our calculation made with their farm information. Underestimating the economic losses of mastitis can be regarded as a general problem in the dairy sector. The average economic losses assessed by the farmers were €78/cow per year, but a large variation was given, €17–198/cow per year. Although the average assessment of the farmers of the different cost factors is close to the default value, there is much variation. To improve the adoption rate of advice and lower the incidence of mastitis, it is important to show the farmers the economic losses of mastitis on their farm. The tool described in this paper can play a role in that process.

Mastitis is an inflammation of the mammary glands and in most cases it is caused by the presence of microorganisms. High mastitis rates in dairy cattle herds can cause an increase in total microorganism counts of bulk tank milk. The present paper was aimed at verifying whether the occurrence of mastitis in dairy cattle herds is reflected in raw-milk indicators of hygienic-sanitary quality. To observe the correlation among the analysed variables, we performed a logarithmical transformation (log10) of different indicator counts of raw milk and compared them with the occurrence of mastitis in dairy cattle herds. Few correlations were observed among mastitis cases in dairy cattle herds and the raw-milk indicators of hygienic-sanitary quality. We observed a negative correlation between the log10 of mesophilic aerobic plate counts and psychotropic aerobic plate counts when compared with the occurrence of no bacterial growth. The log10 of thermophilic aerobic plate counts and yeasts and mould aerobic plate counts presented a positive correlation with the cases of infectious mastitis and mastitis caused by Staphylococcus spp.