Subclinical ketosis is a metabolic disorder which often goes undiagnosed and leads to constricted performance and an impairment of general condition. In the current study subclinical ketosis was characterised by a β-hydroxybutyrate (BHB) concentration of >1·2 mmol/l in blood serum. To generate this metabolic situation, an animal model was created. The model, based on group-specific interaction of dietary energy supply and body condition, is appropriate for testing the medical effectiveness of treating this kind of ketosis and its concomitants. During the trial, 18 dairy cows (primiparous and pluriparous) were assigned, according to their body condition score (BCS) 6 weeks before expected parturition, to a normal [6·78 MJ net energy for lactation (NEL)/kg dry matter; 20% concentrate] or to a high-energy feeding group (7·71 MJ NEL/kg dry matter; 60% concentrate). Therefore cows with the highest BCS were allocated to the high-energy group to enhance the contrast with the control group. Statistical analysis was done using the MIXED procedure of SAS. Effects were declared significant when P-values were ⩽0·05. Owing to the higher energy concentration and dry matter intake, the energy intake and balance was significantly higher in the high-energy feeding group, with strong effects on lipid metabolism and health in blood and liver post partum. Within the first 2 weeks after calving, 8 out of 9 cows (89%) of the high-energy group had BHB values indicative of subclinical ketosis. These cows also had significantly higher values of non-esterified fatty acids (NEFA), aspartate transaminase (AST) and glutamate dehydrogenase (GLDH) post partum, as well as a raised total lipid content of the liver. RQUICKI, a calculated parameter which is based on serum concentrations of glucose, insulin and NEFA to assess the insulin sensitivity, was not affected by treatment. Therefore, RQUICKI does not seem to be the right parameter for diagnosing decreased insulin sensitivity in cows affected by subclinical ketosis. The milk fat and the fat:protein ratio of the high-energy group was also higher, even though there was no decrease in milk yield for cows with subclinical BHB values.

Seven changeover experiments were conducted to examine the effect of giving restricted amounts of concentrates on the voluntary intake of different roughages by non-lactating dairy cows. The addition of up to 6 kg concentrates daily to the diet of cows receiving roughage ad lib. caused little change in the intake of hay and a small increase in the intake of barley straw. Larger amounts of concentrates of 6 and 8 kg daily reduced hay intake by between 0·2 and 0·4 kg dry matter/kg concentrate dry matter given. When concentrates were given to the cows the rate of decline in intake of hay tended to be greatest with the hays of highest digestibility. The daily addition of up to 8 kg concentrates to the diet of cows offered silage ad lib. caused only small depressions in the intake of silage. The results are discussed in relation to recent work on this subject with cattle and sheep.

The objectives of this research were to confirm the effect of compounds derived from milk fermented by Lactobacillus helveticus (LH-2) on the nonspecific host defence system, and isolate and characterize the active peptides that mediate the immune response. The cell-free supernatant obtained from the fermented milk and its fractions were tested in vitro for immuno-modulating activity using murine macrophages (RAW 264·7 cell line). Cytokine production (Interleukin-6 (IL-6), Tumor Necrosis Factor-α (TNF-α), and Interleukin-1β (IL1-β)), nitric oxide (NO) production and phagocytosis were used as biomarkers. Macrophages stimulated with cell-free supernatant of fermented milk showed higher production of cytokines and NO compared with macrophages stimulated with LPS (Lipopolysaccharide) and a commercial immunomodulator derived from β-casein (f54-59). Phagocytosis was observed by macrophages stimulated with the supernatant. Two of nine fractions collected from the supernatant using size exclusion chromatography produced the highest response when used to stimulate macrophages. The results of the dose-response study of the effect of the fraction with the highest stimulation effect on the production of TNF-α showed a direct correlation between protein concentration and TNF-α release. The fraction contained four novel peptides, three derived from the hydrolysis of β-casein and one from the hydrolysis of α-lactalbumin. These results confirm that fermentation of milk by Lactobacillus helveticus (LH-2) results in the production of specific peptides capable of modulating macrophage activity.

In a national survey of US dairy operations, 1685 dairy operations reported 47057 new dairy heifers (either births or acquisitions) and 4427 deaths (9·4%) of preweaned dairy heifer calves over a 3 month period. Stepwise logistic regression was used to identify management practices associated with high mortality among preweaned heifers in dairy operations where at least three dairy heifer calves were born alive or moved on to the operation. Analysis was done twice: once by separating all operations by size into high or low mortality; again using only operations with <2 and >10% mortality to eliminate dairy operations with intermediate levels of mortality from the comparisons. Results were similar. Dairy operations in the West were more likely to fall in the high mortality category than dairy operations in the rest of the country. In addition, the following dairy operation characteristics were associated with high death levels in both models: rolling herd average milk production <7710 kg, preweaned heifers placed in groups of seven or more, a male having primary responsibility for the care and feeding of preweaned heifers, calves not receiving hay or other roughages until >20 d old, calves fed on mastitic or antibiotic milk after colostrum and calves not given whole milk after colostrum.

Agriculture and animal husbandry are important contributors to global emissions of greenhouse (GHG) and acidifying gases. Moreover, they contribute to water pollution and to consumption of non-renewable natural resources such as land and energy. The Life Cycle Assessment (LCA) methodology allows evaluation of the environmental impact of a process from the production of inputs to the final product and to assess simultaneously several environmental impact categories among which GHG emissions, acidification, eutrophication, land use and energy use. The main purpose of this study was to evaluate, using the LCA methodology, the environmental impact of milk production in a sample of 41 intensive Italian dairy farms and to identify, among different farming strategies, those associated with the best environmental performances. The functional unit was 1 kg Fat and Protein Corrected Milk (FPCM). Farms showed characteristics of high production intensity: FPCM, expressed as tonnes per hectare, was 30·8±15·1. Total GHG emission per kg FPCM at farm gate was 1·30±0·19 kg CO2 eq. The main contributors to climate change potential were emissions from barns and manure storage (50·1%) and emissions for production and transportation of purchased feeds (21·2%). Average emission of gases causing acidification to produce 1 kg FPCM was 19·7±3·6 g of SO2 eq. Eutrophication potential was 9·01±1·78 ${\rm PO}_{\rm 4}^{{\rm 3} -} {\rm eq}.$ per kg FPCM on average. Farms from this study needed on average 5·97±1·32 MJ per kg FPCM from non-renewable energy sources. Energy consumption was mainly due to off-farm activities (58%) associated with purchased factors. Land use was 1·51±0·25 m2 per kg FPCM. The farming strategy based on high conversion efficiency at animal level was identified as the most effective to mitigate the environmental impact per kg milk at farm gate, especially in terms of GHG production and non-renewable energy use per kg FPCM.

To evaluate the bioactivity of bovine milk from different stages of lactation on human intestinal tissue, a human fetal small intestinal cell line was used as a model system. Milk samples representing six stages of lactation: days 1, 2–3, 6–7 and weeks 12 and 24 after parturition, 1 week before drying off, and milk-like secretion from two stages of the dry period: 7 weeks and 3–4 weeks before expected calving, were collected from 64 Holstein Friesian cows. The whey fraction of the milk or milk-like secretion was added to the culture medium in concentrations ranging from 0·078% to 10%. The growth-promoting activity of whey was measured by determining the incorporation of [3H]thymidine into DNA for the last 24 h of the culture period. Whey fractions from all six stages of lactation stimulated growth of intestinal cells. The growth-promoting activity of colostrum or milk significantly decreased within the first week after calving. The growth-promoting activity in mature milk increased gradually during lactation to reach a level significantly higher than that obtained with colostrum. The growth-promoting activity of whey from milk-like secretion collected after drying off was lower than that of colostrum. Whey from different stages of lactation contained significantly different concentrations of TGF-β1 (0·5–27 ng/ml) and TGF-β2 (12–1219 ng/ml). However, neither the differences in TGF-β1 and TGF-β2, nor the differences in IGF-I and IGF-binding proteins could fully explain the differences in growth-promoting activity of colostrums or milk from different stages of lactation, suggesting that other factors were also involved. The present study showed that bovine milk contained a number of biologically active components that affected growth and development of human intestinal tissue. The results showed that the growth-promoting activity of colostrum and milk was dependent on the stage of lactation in accordance with previous results obtained with mammary epithelial cells. The changes in growth-promoting activity with stage of lactation were probably related to changes in concentrations of several growth factors.

The sensory and rheological characteristics of stirred yogurts varying in fermentation temperature, heat treatment of milk, dry matter content and composition of bacterial cultures were investigated. Two independent sensory properties, non-oral viscosity and oral viscosity, were evaluated. To reflect these properties precisely it was necessary to perform rheological measurements at both small and large deformations. Non-oral and oral viscosities were successfully modelled from a set of rheological variables by partial least squares regression, resulting in a three component model explaining 83·8% of the variation in non-oral viscosity and a two component model explaining 82·0% of the variation in oral viscosity. Non-oral viscosity was highly correlated with the complex modulus (G*) from the dynamic oscillatory measurements (r=0·823) and the viscosity obtained from a Brookfield viscometer (with Helipath stand) operating at 5 rev./min (r=0·862). Similarly, there were relationships between oral viscosity and the hysteresis loop area from the viscometry test (r=0·867) and between oral viscosity and the viscosity obtained from the Posthumus funnel (r=0·834).

Casein phosphopeptides (CPP) which develop in Grana Padano cheese at different ages were isolated by precipitation with Ba2+ and analysed by HPLC. Profiles were complex throughout the period between 4 and 38 months. CPP in a cheese sample 14 months old were identified by a combination of fast atom bombardment–mass spectrometry and Edman degradation. They were found to consist of a mixture of components derived from three parent peptides, β-CNf(7–28)4P, αs1-CNf(61–79)4P and αs1-CNf(7–21)4P. In total, 45 phosphopeptides were identified: 24 from β-CN, 16 from αs1-CN and 5 from αs2-CN. The presence of aminopeptidase activity during cheese ripening was deduced from the presence of a number of CPP of different lengths with the loss of one or more residues from the N-terminus. The longest had C-terminal lysine and seemed to be progressively hydrolysed by carboxypeptidases A and B to shorter peptides. CPP in cheese appeared to be shortened plasmin-mediated products. Moreover, those most resistant to further hydrolysis contained at least three closely located phosphoserine residues. The anticariogenic activity of CPP is also discussed.

Intracellular dipeptidase of Streptococcus cremoris NCDO 924 has been investigated as a marker enzyme for detecting the release of starter enzymes into Cheddar cheese during maturation. Dipeptidases were produced extra- and intracellularly by Str. cremoris in broth cultures, but the intracellular enzyme was distinguishable by its relatively high activity against L-alanylglycine at low temperatures. The presence of dipeptidase activity of intracellular origin was demonstrated in extracts of Cheddar cheese freed from whole starter cells, opaque material of high molecular weight and free amino acids by a combination of centrifugation and gel filtration on Sephadex G200. The enzyme was not detected in cheeses made without starter (δ-gluconic acid lactone cheeses). In fresh curd with a high viable count of starter the dipeptidase activity was low and mostly extracellular. As the count declined the proportion of intracellular enzyme activity increased, reaching a maximum when 90% of the starter cells had died. The total dipeptidase subsequently decreased until the level remained 30% higher than that of the original curd, and changed little on further ripening up to 120 d. There remained a significant proportion of extracellular dipeptidase activity throughout the maturation period studied.

Manchego cheese can be manufactured from raw or pasteurized ewes' milk. An automatic purge and trap apparatus, coupled to a GC-MS was used to isolate, identify and compare the relative amounts of the volatile components of raw and pasteurized Manchego cheese during ripening. The majority of volatile compounds were more abundant in raw milk (RM) cheeses than in pasteurized milk (PM) cheeses. Alcohols and esters predominated in the profile of RM Manchego cheese, while methyl-ketones and 2,3-butanedione were quantitatively important in PM cheeses. Branched chain alcohols were much more abundant in RM cheeses. The discriminant analysis separated 100% samples into RM or PM cheeses by using only 16 volatile compounds. Aroma intensity was correlated with esters, branched chain aldehydes and branched chain alcohols in RM cheeses, and with esters, branched chain aldehydes, 2-methyl ketones and 2-alkanols in PM cheeses. Diacetyl was positively correlated with the aroma attribute ‘toasted’ and negatively correlated with aroma quality in PM cheeses.

The rates of proteolysis during ripening were followed in cheeses made with either calf rennet or swine pepsin and either starter or δ-gluconic acid lactone (GAL) as a replacement for the starter. A gel-filtration column technique and starch-gel electrophoresis were used for analysis, and bacterial counts were made on all samples. Proteolysis was faster in cheeses made using GAL than in those made using starter and also slightly faster in GAL cheeses made with swine pepsin than in those made with rennet. Further, it was considerably slower in starter-containing cheeses made with swine pepsin than in those made with rennet. It is suggested that these differences were due to the much greater rate of development of acidity in cheeses made with GAL than in those made with starter, which resulted in more of the coagulant being incorporated into the curd in an active state. The rate of proteolysis in starter-containing cheeses appeared to follow a characteristic course, being initially slow, then markedly increasing with a later slow decline. It is suggested that the increase in the rate of proteolysis was due to an increase in the total activity of bacterial proteinases released by lysis of the bacteria. Indications were obtained that the coagulants and bacterial proteinases catalysed broadly similar patterns of protein breakdown in cheese, and that medium-sized peptides (mol. wt 9000–14000) were formed as definite intermediates in the process. The results also showed that rennet and swine pepsin remained active for at least 7 months in GAL cheeses, that rennet contributed significantly to proteolysis in starter-containing cheeses, and that swine pepsin was at least extensively inactivated and possibly completely inactivated during cheese-making with starter.

The aim of the study was to investigate the effects of season, cow cleanliness and milking routine on bacterial and somatic cell counts of bulk tank milk. A total of 22 dairy farms in Lombardy (Italy) were visited three times in a year in different seasons. During each visit, samples of bulk tank milk were taken for bacterial and somatic cell counts; swabs from the teat surface of a group of cows were collected after teat cleaning and before milking. Cow cleanliness was assessed by scoring udder, flanks and legs of all milking cows using a 4-point scale system. Season affected cow cleanliness with a significantly higher percentage of non-clean (NC) cows during Cold compared with Mild season. Standard plate count (SPC), laboratory pasteurization count (LPC), coliform count (CC) and somatic cell count, expressed as linear score (LS), in milk significantly increased in Hot compared with Cold season. Coagulase-positive staphylococci on teat swabs showed higher counts in Cold season in comparison with the other ones. The effect of cow cleanliness was significant for SPC, psychrotrophic bacterial count (PBC), CC and Escherichia coli in bulk tank milk. Somatic cell count showed a relationship with udder hygiene score. Milking operation routine strongly affected bacterial counts and LS of bulk tank milk: farms that accomplished a comprehensive milking scheme including two or more operations among forestripping, pre-dipping and post-dipping had lower teat contamination and lower milk SPC, PBC, LPC, CC and LS than farms that did not carry out any operation.

Additions of potassium iodate to milk at 0·05 and 0·1 mM (10 and 20 ppm) before UHT treatment markedly reduced the rate at which pressure built up during processing. This permitted the use of longer processing times before unacceptable pressures were reached in the heat exchangers. Iodate reduced the amount of protein deposited, particularly in the higher temperature sections of the plant, but had no effect on the deposition of minerals. The more compact nature of the highly mineral deposits offered less resistance to the flow path. Reduction in the amount of protein deposited is likely to be caused by increased denaturation of β-lactoglobulin and oxidation of heat activated sulphydryl groups by the iodate, thus reducing the formation of high molecular weight polymers of sulphur-containing proteins at the heated surfaces. Increasing the level of sulphydryl groups in the milk through the addition of L-cysteine-HCl caused an increase in the amount of deposit formed during UHT treatment. Whilst little detrimental effect on the quality of the milk resulted from additions of iodate at 0·05 mM, milks with 01 mM-iodate became bitter during subsequent aseptic storage. Bitterness was a result of iodate-induced proteolysis of casein.

The influence of diet quality and stage of lactation on Cheddar cheese moisture and yield were segregated and the effects of other potential variables (cow breed, milk protein genotype, age and level of mastitis) were minimized. An experiment was conducted using milk of cows in mid lactation or late lactation fed on high or low quality diets. Increased moisture in cheese was largely due to the influence of cow diet. Milk from cows given high quality diets produced lower moisture cheese. Cheese yield was influenced by diet and stage of lactation. Cows in late lactation fed on high quality diets always gave milks yielding the most cheese. Fat and protein recovery in cheese was not influenced by diet or stage of lactation. The results suggest that low quality diets may induce changes in the complex interactions of milk proteins and other components that affect cheesemaking.

After extended tryptic hydrolysis of large bovine casein micelles, a mineral-rich peptide fraction was recovered by ultracentrifugation. Its mineral part contained 72% of the colloidal Ca and 49% of the colloidal Pi originally present in the native micelle. Colloidal nitrogenous components were also present, amounting to 27% of the original N content. They contained most of the phosphopeptides and 82% of the micellar phosphoseryl residues. These tryptic peptides were characterized by reversed-phase HPLC on-line electrospray ion source–mass spectrometry analysis. Among the peptides produced 14 phosphopeptides were identified: αs2-CN(l–24), αs2-CN(1–21), αs1-CN(43–79), αs1-CN(35–79)7P, αs1-CN(35–79)8P, αs1-CN(37–79), αs1-CN(104–119), αs1-CN(104–124), β-CN(1–25), β-CN(1–28), β-CN(1–29), β-CN(30–97), β-CN(33–97) and β-CN(29–97). The proportion of the phosphopeptides interacting with colloidal calcium phosphate was correlated with their relative content of phosphoserine residues, since phosphopeptides containing more than four phos-phoserine residues were consistently present within this fraction. It also appeared that other types of peptides, some of them hydrophobic in nature, were also partly or completely present within the colloidal fraction, including αs1-CN(91–100), αs1-CN(152–193), αs1-CN(23–34), αs1-CN(125–193), αs1-CN(125–199), β-CN(177–209), β-CN( 184–209), β-CN(114–169) and β-CN(108–169). Their possible involvement in the micellar backbone is discussed.

The effects of the isocaloric replacement of part of the dietary concentrate mixture by 5% lauric acid (99% pure) or by 10% of myristic (95% pure), palmitic (96% pure) or stearic (94% pure) on the yield and composition of milk fat was investigated in 2 feeding experiments with a total of 10 cows in mid-lactation. The concentrate mixtures were given with a high-roughage diet that supplied 9·1 or 7·7 kg hay/day in expts 1 and 2, respectively.

In expt 1 the inclusion of myristic acid in the diet decreased the yields of milk and solids-not-fat (SNF), but increased the percentage of fat in the milk without altering the fat yield. When stearic or palmitic acid was included in the diet there was an increase in milk fat yield; palmitic acid caused the greater increase. In expt 2 the inclusion of lauric acid in the diet did not appear to affect the yields of milk or SNF but it caused large reductions in the percentage of fat in the milk and in the yield of fat.

In expt 1 the inclusion of myristic acid increased the yields and percentages of 14:0, 14:1 and 16:1 in the milk fat and decreased the yields and percentages of the short-chain fatty acids (4:0–8:0), 16:0, 18:0 and 18:1. When palmitic acid was included in the diet there were increases in the yields and percentages of 16:0 and 16:1 in the milk fat and decreases in the yields and percentages of 10:0, 12:0, 14:0, 18:0 and 18:1. The inclusion of stearic acid in the diet increased the yields and percentages of 18:0 and 18:1 in the milk fat and decreased the yields and percentages of 12:0 and 16:0. In expt 2 the inclusion of lauric acid in the diet increased the yield and percentage of 12:0, and in 1 cow the content of 14:0 in the milk fat; there were reductions in the yields and percentages of all the other fatty acids in the milk fat. There was no evidence of any marked elongation of the carbon chains of 12:0 and 14:0 in the mammary gland to form 16:0 or 18:0.

In expt 1 the incorporation of myristic acid in the diet decreased the concentration of total steam-volatile fatty acids in rumen liquor. The inclusion of either myristic, palmitic or stearic acid in the diet had little effect on the relative proportions of the individual volatile acids in the rumen liquor. In expt 2 the addition of lauric acid to the diet reduced the acetic acid:propionic acid ratio in the rumen liquor.

The apparent digestibility coefficients of the various constituents of diets containing myristic, palmitic or stearic acids have been determined in an experiment with 8 sheep.

The inclusion of myristic acid in the diet had no effect on the digestibility coefficient of total dry matter or crude protein, but it decreased the digestibility coefficients of crude fibre and nitrogen-free-extractives and increased the digestibility coefficients of fat and minerals. When palmitic acid was included in the diet there were decreases in the digestibility coefficients of total dry matter, crude fibre and nitrogen-free-extractives. The inclusion of stearic acid in the diet resulted in decreases in the digestibility coefficients of dry matter and nitrogen-free-extractives. There was an inverse relationship between the melting points and the digestibility coefficients of the added fatty acids.

The findings of this investigation are discussed in relation to the results of a previous study on the effects of these dietary fatty acids on the yield and composition of cow's milk.

The acid-soluble ribonucleotides of human milk were measured by enzymic and ion-exchange chromatographic procedures at different stages of lactation. Human colostrum and milk contained CMP, AMP, GMP, UMP, GDP-mannose, UDP-N-acetyl-glucosamine, UDP-N-acetyl-galactosamine, UDP-glucose, UDP-galactose, UDP and other minor nucleotides. Cytidine and adenosine derivatives were present in relatively higher amounts than in milk from ruminant species. The nucleotide concentration decreased with advancing lactation. Human milk at 3 months of lactation contained about 10 µmol/100 ml nucleotides, which represented about 75% of the amount of nucleotides present in human colostrum. Human milk did not contain orotic acid. The UDP-glucose/UDP-galactose ratio was constant during lactation and was similar to that of milks from ruminant species.

Sub-clinical mastitis (SCM) affects milk composition. In this study, we hypothesise that large-scale mining of milk composition features by pattern recognition models can identify the best predictors of SCM within the milk composition features. To this end, using data mining algorithms, we conducted a large-scale and longitudinal study to evaluate the ability of various milk production parameters as indicators of SCM. SCM is the most prevalent disease of dairy cattle, causing substantial economic loss for the dairy industry. Developing new techniques to diagnose SCM in its early stages improves herd health and is of great importance. Test-day Somatic Cell Count (SCC) is the most common indicator of SCM and the primary mastitis surveillance approach worldwide. However, test-day SCC fluctuates widely between days, causing major concerns for its reliability. Consequently, there would be great benefit to identifying additional efficient indicators from large-scale and longitudinal studies. With this intent, data was collected at every milking (twice per day) for a period of 2 months from a single farm using in-line electronic equipment (346 248 records in total). The following data were analysed: milk volume, protein concentration, lactose concentration, electrical conductivity (EC), milking time and peak flow. Three SCC cut-offs were used to estimate the prevalence of SCM: Australian ≥ 250 000 cells/ml, European ≥200 000 cells/ml and New Zealand ≥ 150 000 cells/ml. At first, 10 different Attribute Weighting Algorithms (AWM) were applied to the data. In the absence of SCC, lactose concentration featured as the most important variable, followed by EC. For the first time, using attribute weighted modelling, we showed that the concentration of lactose in milk can be used as a strong indicator of SCM. The development of machine-learning expert systems using two or more milk variables (such as lactose concentration and EC) may produce a predictive pattern for early SCM detection.