Two experiments were undertaken to determine the effects of cereal grain and fibre (hay or straw) supplements on the fatty acid composition of milk fat of grazing dairy cows in early lactation. In both experiments, grain supplements significantly increased (P<0·05) the proportion of the endogenously synthesized 10[ratio ]0–16[ratio ]0 fatty acids. Of the C18 acids, the proportion of 18[ratio ]0 and 18[ratio ]3 was significantly decreased (P<0·05) by grain supplementation, while that of 18[ratio ]2 was significantly increased (P<0·05). Irrespective of diet, 18[ratio ]1 trans-11 was the most dominant trans 18[ratio ]1 isomer in milk fat. In the first experiment, the proportions of the 18[ratio ]1 trans-11 isomer and conjugated linoleic acid (CLA, 18[ratio ]2 cis-9, trans-11) were highest for the pasture-only diets, and significantly (P<0·05) decreased with grain supplementation. The opposite result was observed in the second experiment, conducted in a different dairy region, suggesting that factors such as the quality of pasture on offer and the physiological state of the cow could affect the content of CLA and trans fatty acids in milk fat. In both experiments, there was a significant positive linear relationship between CLA and 18[ratio ]1 trans-11. Fibre supplements had little effect on the fatty acid composition of the milk.

In five short-term experiments conducted in Victoria in 1997 and 1998, grazing dairy cows were given either pasture alone or pasture supplemented with high-energy concentrates, and the fatty acid profiles of milk fat were measured. We established the effects of these feeds on some aspects of milk fat of importance for human nutrition, but we specifically focused on the hypothesis that conjugated linoleic acid (CLA) concentrations in milk fat increase as pasture intake increases, and decrease as more concentrates are fed. In agreement with previous research, feeding fresh pasture alone resulted in high concentrations (1·0–1·8 g/100 g milk fat) of CLA. When the effect of level of pasture consumption on CLA content was examined, a significant positive relationship (r2=0·35; P<0·05) was obtained. When cereal grain concentrates were used to supplement pasture intake, the CLA content of milk fat generally declined (P<0·05), except where the amount of concentrates given led to a marked reduction in total milk fat concentration. The use of cereal grain concentrates also generally resulted in significant (P<0·05) increases in medium-chain saturated fatty acids, but always reduced the contribution of butyric acid to milk fat, from 4·5 to 3·9 g/100 g milk fat, on average.

The aim of the study was to determine the role of opioids modulating the release of prolactin (PRL) in response to milking in sixteen Brown-Swiss dairy cows. Two experiments were carried out to measure the dose-related effect of morphine and the effect of the opioid antagonist naloxone (NAL), with or without morphine. In the first experiment, six cows were injected (via catheter) on 3 consecutive days after the control milking (0 mg) with 21, 70 and 210 mg morphine-HCl 10 min before the start of the morning milking. The second experiment was divided into two parts. In the first part, four cows were injected after control morning milking with 210 mg morphine, 10 min before the start of the following morning milking. This was followed on the next day by an application of 210 mg NAL (15 min before the start of milking) and 210 mg morphine. In the second part, after control milking for 1 d, six cows were injected with 210 mg NAL 10 min before milking. Morphine at the highest dose tended to stimulate basal PRL levels in the first and second experiments (P<0·10). PRL increased in response to machine milking but morphine did not stimulate its release further. NAL alone, or when given with morphine did not influence the release of PRL in response to machine milking. However, NAL was effective in suppressing the increase in basal levels of PRL caused by morphine. In conclusion, although morphine tended to stimulate basal levels of PRL before milking, the release of PRL during milking seemed not to be regulated by opioids.

The vacuum in the mouthpiece chamber (MPC) was measured in a factorial trial with two liners (DeLaval 964008-01 and DeLaval 999007-03), two pulsator ratios (60 and 70%) and morning and evening milking over eight milkings of ten cows. Teat length and diameter were measured before and after milking, and maximum milk flow rate and depth of penetration of the teat into the teatcup were also recorded. The MPC vacuum at peak flow rate was classified as stable, at various levels, in 79% of the recordings. In the rest of the milkings the vacuum declined gradually, and this decline was associated with gradually deeper penetration of the teat into the teatcup. The DeLaval 964008-01 liner with the narrower bore and larger mouthpiece opening gave lower MPC vacuum at peak flow rate than did the wider bore DeLaval 999007-03 with the smaller mouthpiece opening. Morning milking gave higher MPC vacuum than evening milking. Increased maximum milk flow rate and increased length and diameter of the teat gave decreased MPC vacuum. The teats were longer before milking in the morning than in the evening. However, during peak flow rate, the teats had penetrated deeper into the teatcup in the evening than in the morning. A marked change in MPC vacuum in the transition to the period of flow rate can be used to identify the start of over-milking of an individual teat. A general model explaining the MPC vacuum as the outcome of the balance between air leakage past the mouthpiece lip and the leakage past the teat in the liner barrel is suggested.

Alpha (α)- and beta (β)-adrenergic receptors in the bovine mammary gland are mainly present in the teat muscles and in the region where large milk ducts reach the cisternal cavities. The aim of the study was to test the hypothesis that the region of the large mammary ducts is the most important location of α- and β-adrenergic receptor stimulation affecting milk ejection and milk removal. Effects of α- and β-adrenergic receptor stimulation and of oxytocin (OT) receptor blockade on milking characteristics were tested in six cows. Milk flow was measured before and after the distal part of one teat, including the teat canal and teat sphincter, had been partly amputated. Before the operation, milk yield and peak flow rate decreased during α-adrenergic receptor stimulation and during the OT receptor blockade, and increased during β-adrenergic stimulation. After removal of the teat tip, relations of milk yield and peak flow rates after administration of α- and β-agonists and after application of an OT receptor blocking agent were similar to those before operation. Only total milk yield had decreased in the teat-amputated quarter owing to unhindered flow of cisternal milk before cluster attachment. Since responses to α- and β-adrenergic receptor stimulation as well as to OT receptor blockade do not differ with or without the teat sphincter, it is concluded that milk flow is mainly influenced by the muscle tone of the large mammary ducts.

Psychrotrophs, particularly Pseudomonas spp. are known to be the main determinants of the shelf-life of pasteurized milk and refrigerated raw milk. It is presumed that they mainly cause spoilage through the elaboration of proteinase and lipase enzymes. At the time of this research, under the relevant European Directive, one of the means of determining the quality of pasteurized milk was the pre-incubated count, which involves incubating the milk sample for 5 d at 6°C followed by a plate count. Examination of numerous pre-incubated counts revealed a bimodal rather than a normal distribution indicating that the types of contaminants in pasteurized milk may be as important as their initial concentration. Pseudomonads that gave particularly high (>5×106 cfu/ml) and low (<103 cfu/ml) pre-incubated counts were isolated (high and low count isolates respectively). After the organisms had been subjected to a cold shock no consistent trend between the groups of isolates was detected with respect to lag phase duration. However, the high count isolates consistently had a faster exponential growth rate. Unexpectedly, with the exception of one isolate, the low count isolates produced detectable proteinase and lipase earlier. In addition, with one exception, maximal proteinase and lipase production was observed with the low count isolates. These findings indicate that there is no causal relationship between selective growth advantage and ability to produce proteinase and lipase. It also indicates that the spoilage of pasteurized milk is a complex phenomenon and is worthy of further research.

Although widely adopted by the chemical and pharmaceutical industries in recent years, little published data is available regarding possible applications of high pressure homogenisation for dairy products. The objective of this work was to compare the effects of conventional (18 MPa, two-stage) and single or two-stage high pressure homogenisation (HPH) at 50–200 MPa on some properties of raw whole bovine milk (∼4% fat). Fat globule size decreased as HPH pressure increased and, under certain conditions of temperature and pressure, HPH yielded significantly smaller fat globules than conventional homogenisation. Fat globule size was also affected by milk inlet temperature. The pH of all homogenised milk samples decreased during 24 h refrigerated storage. Total bacterial counts of milk were decreased significantly (P<0·05) for milk samples HPH-treated at 150 or 200 MPa. Whiteness and rennet coagulation properties of milk were unaffected or enhanced, respectively, as homogenisation pressure was increased. Average casein micelle size decreased slightly when skim milk was homogenised at 200 MPa. Thus, HPH treatment has several, potentially significant, effects on milk properties.

The objective of this study was to determine the effect of high pressure homogenisation (HPH) on alkaline phosphatase activity and plasmin and plasminogen-derived activities in raw whole bovine milk. Milk (approximately 4% fat) was treated by two-stage conventional homogenisation (18 MPa) or single or two-stage HPH at 50, 100, 150 or 200 MPa. Inactivation of plasmin and plasminogen-derived activities was evident in conventionally homogenised samples, and increased as HPH pressure increased. Two-stage HPH reduced both activities to a greater extent than single-stage HPH. Milk inlet temperature had a significant effect on residual plasmin and plasminogen activities of HPH-treated milk samples, especially those treated at 50 MPa. Inactivation of plasmin and plasminogen on HPH-treatment (150 MPa) of milk samples of varying fat contents (0–10%) was also investigated; there was a curvilinear relationship between residual plasmin and plasminogen-derived activities and fat content in the ranges 0–2% and 0–4%, respectively, with little additional inactivation at higher fat contents. Thus, indigenous proteolytic activity of milk is clearly affected by HPH. However, all homogenised milk samples retained active alkaline phosphatase, indicating that thermal conditions during HPH did not equate to that of conventional high temperature short time pasteurisation, and that the wide range of forces experienced by milk during HPH treatment does not inactivate the latter enzyme.

Four Lactobacillus helveticus strains were studied for proteolytic capacity and general aminopeptidase (AP) and X-Pro dipeptidyl aminopeptidase (DAP) activity. The rate of hydrolysis and the activity against synthetic substrates with N-terminal residues of Arg, Lys, Leu, Glu or Pro, varied markedly among the strains. The X-Pro DAP activity was consistently high. The crude cell-wall and cytoplasm extracts from strain Lb. helveticus ISLC59 were analysed thoroughly for their proteolysis ability by using four synthetic peptide substrates, including αs1-CN(f1-23). Peptides formed during in vitro hydrolysis of the synthetic substrates by cell wall and cytoplasm preparations were identified by LC-ESI/MS. In doing so, it was possible to infer a prevalent endopeptidase activity splitting Lys7-His8 and Gln13-Glu14 bonds in the cytoplasm, and to deduce a secondary activity, which hydrolysed Glu14-Val15, Leu16-Asn17, Glu18-Asn19 and Lys3-His4 bonds lacking in the cell-wall. The presence of exopeptidases, as mainly AP, DAP, and carboxypeptidase (CPase) was deduced from the formation of several N- and C-terminally truncated peptides sets. The AP activity was higher in the cell-wall layer, where CPase activity was absent. The in vitro assays with cell extracts of the Lb. helveticus ISLC59 strain revealed extensive exopeptidase and endopeptidase activities. In several cases, the hydrolytic system of Lb. helveticus that splits in vitro αs1-CN(f1-23) peptide bonds was similar to that of Lactococcus lactis. The effects were also compared with those occurring in vivo in hard cheese such as Grana Padano.

A new method for characterization of acid production by dairy starter cultures is presented. Microplates with integrated optical pH sensors are developed. Two fluorophores, a pH-sensitive and a pH-insensitive one are immobilised at the bottom of a polystyrene 96-well microtitre plate. The pH-insensitive fluorophore serves as an internal reference and makes calibration unnecessary. The sensor measures pH accurately in optically well-defined media. Particles and fluorophores contained in the bulk medium disturbed the measurements. Despite these disturbances it was possible to clearly sense differences in inoculum type and in inoculum sizes of cultures of Lactococcus lactis and of Streptococcus thermophilus at 30 and 37°C. Besides a pH-related signal there is information about other changes during milk fermentation. The cultivation results were compared with those from the established CINAC-method. From this comparison it can be concluded that the new method can be used reliably to characterize particularly a large number of strains for screening purposes but also for quality control.

A kinetic model was proposed for describing the curd assembly of skimmed goats' milk during enzymic coagulation. The enzymic coagulation of milk was monitored using an optical sensor that measured diffuse reflectance (light backscatter) at 880 nm. The appearance of a shoulder, at low temperatures and protein concentrations, in the diffuse reflectance ratio profile after the inflection point of the curve (Tmax) appeared to separate the aggregation and curd firming steps. The diffuse reflectance ratio profile after Tmax was attributed to the overlapping of casein micelles aggregation and curd firming reactions. The developed kinetic model combined a second order reaction model to describe aggregation reactions and a first order reaction model to describe firming processes reactions. A completely randomised block design with three replications was used to determine the effect of protein concentration and temperature on kinetic constants. Milk was adjusted to three levels of protein (30, 50 and 70 g/kg), and coagulated at five temperatures (20, 25, 30, 35 and 40°C) to test a wide range of processing conditions. Data points from each profile after Tmax were fitted to the proposed model using non-linear regression. The average R2 and standard error of prediction (SEP) for 45 tests conducted were in the range of 0·9975±0·0027 and 0·0081±0·0037, respectively. A significant increase in characteristic times for aggregation (τ2) and curd firming processes (τ1) were found when temperature decreased or protein increased. Theoretical asymptotic value of reflectance ratio, R∞, increased with increasing level of protein and temperature (P<0·05). The parameter β1, which represented the fraction of diffuse reflectance ratio attributed to aggregation, decreased with increasing temperature and decreasing protein.

Release of oxytocin (OT) is essential for milk ejection in dairy cows (Lefcourt & Akers, 1983; Bruckmaier & Blum, 1998). During milk ejection, alveolar milk is shifted into the cistern, which causes an increase of intracisternal pressure (Bruckmaier et al. 1994). To initiate maximum milk ejection at the start of milking, increasing OT concentration beyond a threshold level is sufficient (Schams et al. 1983). Increasing OT concentration beyond this threshold has no additional effect on intracisternal pressure, i.e., milk ejection (Bruckmaier et al. 1994). Stimulatory effects of milking by hand or by machine or by suckling are well documented (Gorewit et al. 1992; Bar-Peled et al. 1995; Tancin et al. 1995; Bruckmaier & Blum, 1996). At the start of milking, stimulatory effects of machine milking without pre-stimulation or with a manual pre-stimulation and subsequent machine milking cause the release of comparable amounts of OT (Gorewit & Gassman, 1985; Mayer et al. 1985; Bruckmaier & Blum, 1996), whereas the timing of the applied pre-stimulation is important for the shape of the milk flow curve. Should the pre-stimulation period be too short, or absent altogether, the start of the main milk flow is delayed resulting in a bimodal milk flow profile (Bruckmaier & Blum, 1996). Furthermore, the stimulation of only one teat causes an OT release similar to that caused by stimulation of all four teats (Bruckmaier et al. 2001). However, milk production is greater for hand milking or suckling than for machine milking, possibly owing to higher OT concentrations (Gorewit et al. 1992; Bar-Peled et al. 1995).

Alpha-lactalbumin (α-la), a calcium metalloprotein, is one of the major serum-proteins in ruminant milk (Jenness, 1982) and induces lactose synthesis in the mammary gland by interacting with the enzyme UDP-galactosyltransferase, giving rise to the heterodimer enzyme lactose synthase (Ebner & Brodbeck, 1968; Kuhn, 1983). The goat α-la transcription unit (LALBA), located on chromosome 5 (Hayes et al. 1993), is organized in 4 exons varying in length from 75 nucleotides (3rd exon) to 329 nucleotides (4th exon) coding for a 123-amino acid polypeptide chain (Vilotte et al. 1991). According to the strong similarity between bovine α-la (Vilotte et al. 1987) and human lysozyme (similar molecular weight, the same number of S-S bonds, identical N and C terminal residues; Peters et al. 1989), it has been proposed that both genes arose from a common ancestor (Vilotte et al. 1991).

A fruity flavour note is traditionally regarded as a defect in cheese varieties such as Cheddar (Bills et al. 1965; McGugan et al. 1975; Horwood et al. 1987). However, fruitiness is an attribute of other cheese varieties such as Parmesan and Parmigiano Reggiano (Dumont et al. 1974; Meinhart & Schreier, 1986). It is well accepted that esters such as ethyl butanoate and ethyl hexanoate cause the fruity flavour described as apple-like or pineapple-like in raw milk and cheeses (Bills et al. 1965; Engels et al. 1997; Friedrich & Acree, 1998). The development of fruity flavour is often attributed to the esterification of free fatty acids and ethanol by esterases from lactic acid bacteria and psychrotrophic pseudomonads (Hosono et al. 1974; Morgan, 1976).