The contribution of the alternative pathway of complement activation to the capacity of normal milk to deposit C3 fragments on bacteria was tested by attempting to block C3 deposition with antibodies to the alternative pathway component factor B (fB). Factor B was purified and antibodies of the IgY class, which does not activate mammalian complement, were obtained from the egg yolk of immunized laying hens. These antibodies specifically inhibited the deposition of C3. This inhibition and the absence of deposition of C4 demonstrated that C3 deposition in normal milk resulted from the activation of the alternative pathway. Antibodies raised in rabbit were used to develop an ELISA for measuring fB concentrations in milk. The mean concentration of fB was 2·06 μg/ml (±0·18, SEM), 0·57% of the mean value found in serum (360 μg/ml). This proportion was comparable to that of serum albumin (0·63% of serum value) but less than the proportion of C3 in milk (2·71%). Nevertheless, fB was apparently not a limiting factor for the functioning of the alternative pathway, since addition of purified fB to normal milk did not improve C3 deposition. In serum, mild heat-treatment (56 °C for 3 min or 50 °C for 45 min) blocked the alternative pathway and destroyed fB, as shown by loss of antigenicity in ELISA. In milk, mild heat-treatment did not abrogate C3 deposition, and fB was protected, retaining its functionality and antigenicity. Heating at 56 °C for at least 45 min was necessary to completely inhibit C3 deposition in normal milk.

Secretory IgA found in external secretions are constituted by polymeric IgA (pIgA) bound to the extra-cellular part of the polymeric immunoglobulin receptor (pIgR). The receptor mediates transcytosis of pIgA across epithelial cells. The aim of the present study was to analyse the evolution of pIgR expression in the sheep mammary gland during the development of the mammary gland and to analyse its hormonal regulation. Gene expression of the pIgR was analysed in sheep mammary gland during pregnancy and lactation. By Northern Blot analysis, we observed that low levels of pIgR mRNA are expressed until day 70 of pregnancy. Accumulation of pIgR mRNA started during the third part of pregnancy and intensified 3 d after parturition to reach highest levels during established lactation (day 70). In situ hybridization analysis was used to confirm the increase in pIgR gene expression per mammary epithelial cell. In order to examine the hormonal regulation of the pIgR expression, virgin ewes were hormonally treated. Treatment with oestradiol and progesterone increased pIgR mRNA levels slightly. Subsequent addition of glucocorticoids induced a significant accumulation of pIgR mRNA in the mammary gland of the treated animals. Immunohistochemical analysis was performed to verify that the increase of pIgR mRNA level was associated with enhancement of the pIgR protein in mammary cells. No increase of pIgR mRNA levels were observed if PRL secretion was blocked by bromocryptine injections throughout the hormonal procedure. In conclusion, the present experiments suggest that the enhancement of pIgR levels during lactation result from combined effects of both prolactin and glucocorticoids.

A nephelometric immunoassay, with a detection range of 0·3 to 5 g IgG1/l, was developed for the determination of immunoglobulin in bovine milk. The assay exhibited no significant cross-reactivity with αS1-casein, αS2-casein, β-casein, κ-casein or β-lactoglobulin and 39% cross-reactivity with IgG2. The nephelometric assay was compared with ELISA and RID (24 h and 48 h incubations) assays using 105 duplicate milk samples covering IgG1 values ranging from 0·45 to 1·8 g1. The results obtained from all assays showed good agreement with the exception of those obtained by the RID assay (24 h incubation) which gave lower results in samples containing more than 1·2 g IgG1/l. It was concluded that the nephelometric assay is a reliable, rapid and convenient method suitable for the quantification of IgG1 in milk. The assay can be configured for routine high-throughput milk quality assurance for IgG1 in dairy laboratories.

The decline in milk yield after peak lactation in dairy animals has long been a biological conundrum for the mammary biologist, as well as a cause of considerable lost income for the dairy farmer. Recent advances in understanding the control of the mammary cell population now offer new insights on the former, and a potential means of alleviating the latter. The weight of evidence now indicates that a change in mammary cell number, the result of an imbalance between cell proliferation and cell removal, is a principal cause of declining production. Further, it suggests that the persistency of lactation, the rate of decline in milk yield with stage of lactation, is strongly influenced by the rate of cell death by apoptosis in the lactating gland. Mammary apoptosis was first demonstrated during tissue involution after lactation, but has now been detected during lactation, in mammary tissue of lactating mice, goats and cattle. Those factors that determine the rate of cell death by apoptosis are as yet poorly characterized, but include the frequency of milking in lactating goats. Initial evidence suggests that nutrition also is likely to influence cell survival after peak lactation, an important factor being the degree of oxidative stress imposed by feed and the tissue's ability to deal with, and prevent damage by, reactive oxygen species. Comparison of cows in calf or not pregnant during declining lactation also indicates a likely influence of reproductive hormones, with oestradiol and progesterone acting to preserve mammary ductal and alveolar integrity during the dry period, while allowing a degree of apoptosis and cell replacement. In each case, the molecular mechanisms controlling mammary cell survival (or otherwise) are as yet poorly defined. On the other hand, more persistent lactations are likely to benefit animal welfare through fewer calvings and by placing less emphasis on maximal production at peak lactation, and modelling of persistent lactation with longer calving intervals indicates their likely economic benefits. In these circumstances, there is considerable incentive to elucidate the determinants of mammary apoptosis, and the factors controlling the dynamic balance between cell proliferation and cell death in the lactating mammary gland.

A total of 7492 test-day observations for mean contents of fat, protein, casein, serum protein and lactose and individual laboratory cheese yield (ILCY) were obtained, at approximately monthly intervals, from 1119 ewes belonging to eight Churra dairy flocks. The effect of various factors on these variables was examined and phenotypic correlations among all traits were estimated. Least squares analyses showed significant effects of flock test-date, stage of lactation, age of ewe, and number of lambs weaned on almost all variables. Protein content and composition were not affected by the number of lambs weaned. ILCY had an unadjusted mean (26·55 kg cheese/100 l milk) close to those reported for real cheese yield in dairy ewes and was affected similarly to the main milk components. Fat, protein, casein, and serum protein contents, and ILCY, showed a generally increasing trend as lactation progressed. These components reached a minimum at 1 month into lactation, when milk yield was highest, and increased for the remainder of the lactation. ILCY depended mainly on fat, protein and casein contents. Protein and casein contents were closely related and equally correlated with ILCY. An increase in somatic cell count (SCC) was associated with decreased milk yield and decreased lactose content.

The aim of this study was to test if the opioid antagonist naloxone has a beneficial effect on normalization of oxytocin (OT) release during repeated milking of cows in unfamiliar surroundings. One control milking without naloxone treatment in all cows was performed in the familiar parlour. For four successive evening milkings, cows were transported to, and milked in, the operating theatre of the research station without (control group) or with naloxone administration (1 mg/kg BW) (naloxone group) before milking. After cessation of spontaneous milk flow, but not before 3 min of milking, vaginal stimulation was applied for 2 min. After milk flow ceased again, 10 IU of OT was injected intravenously to remove the remaining milk including residual milk. Milk flow was recorded continuously and blood samples were collected via a jugular vein cannula at 1-min intervals from 1 min before the start of milking until i.v. injection of OT. The inhibition of milk ejection and its normalization during repeated milking in unfamiliar surroundings was not influenced by naloxone treatment. Concentrations of cortisol and β-endorphin during control milking and all relocations were similar in the naloxone and control groups, although their concentrations were higher after relocations than in the control. Therefore, a role of endogenous opioids in the inhibition of milk ejection in unfamiliar surroundings could not be demonstrated. In addition, the effect of exogenous ACTH1–24 (8 IU, i.v.) on the release of cortisol related to the response of cows milked in unfamiliar surroundings was studied. Cows with totally inhibited milk ejection in response to vaginal stimulation during milking after first relocation had numerically, but not significantly lower cortisol levels (8·8±3·4 ng/ml; AUC/min) in response to ACTH than did cows with at least partial milk ejection (38·7±12·9 ng/ml). Thus animals with a higher adrenal response to ACTH seemed to have less severe inhibition of milk ejection.

The International Standards Organization (ISO) produces international standards for milking equipment. These describe the minimum specifications of design, installation, maintenance and testing of milking machines (BS ISO, 1996). Often these standards are adopted and exceeded by national standards. Standards are rarely mandatory and in some countries variations in interpretation and even disagreement can lead to differences in operating conditions of the milking machine.

A survey of plant test results in the UK (Berry & Scrivens, 1997) showed that, on annual or biannual testing, more than 75% of plants failed to meet the then existing standard (BS5545, 1988). In the UK, most types of plant installed since 1994 meet the newest ISO standard (BS ISO, 1996). However, one type of plant has many features different from all other types. This is a type of plant whose major variations include a smaller claw bowl volume (150 ml), a narrower long milk tube diameter (13·5 mm), 8 mm milk pulse tube, greater cluster weight (approximately 3·5 kg) and simultaneous pulsation used on a plant with milk lift and milking at an installed system vacuum of 47–48 kPa. The majority of parlours installed with this equipment at the time of this study had no form of indexing (see below).

A study has been undertaken to assess 20 new installations for compliance with the ISO requirements (BS ISO, 1996), performance in a static test, milking performance, cow behaviour and the effects on teat condition. Compliance to standards and static test results have been reported elsewhere (Ohnstad, 1997), with no plant proving entirely satisfactory. Some plants had major problems with vacuum level, pulsation characteristics and large air leaks. The physical operation of the milking plant has important implications for cow behaviour and welfare and also ergonomics of the milking operation. The effects on teat condition have already been reported (Hillerton et al. 2000). Here the effects on cow behaviour and milking performance are assessed. Comparisons have been made in line with the International Dairy Federation recommendations (IDF, 1997) to examine the performance and interactions of the cow, the machine and the operator in evaluating the milking process.

Unsatisfactory milking conditions, including high milking vacuum, ineffective pulsation, heavy clusters, unsuitable liners, poor teat preparation and over-milking, are known to create poor, but undefined, teat conditions and pose a risk to an increased probability of intramammary infection (IDF, 1994). It has been shown that poor teat condition, seen as discolouration, abnormal firmness or thickened rings of tissue distally, after cluster removal may be common in a variety of commercial milking operations (Hillerton et al. 2000). That field study was observational and teat conditions were influenced by a number of variables. A small experimental study has been undertaken to develop an experimental model, under controlled milking conditions, to allow investigation of potential causes of teat trauma. Over-milking was suggested as highly important in the field observations and so has been examined in this study.

Samples of bovine caseinomacropeptide (CMP) were isolated from κ-casein A and κ-casein B and fractionated to give aglycosylated CMP A and CMP B and monoglycosylated CMP A. The secondary structures of these three peptides were compared under neutral and acidic (pH 4·2) conditions, using two-dimensional (2D) 1H nuclear magnetic resonance (NMR) spectroscopy. The differences between the spectra at pH 4·2 and 7·0 and the spectra of the aglycosylated and glycosylated CMP A were subtle, indicating little change in backbone conformation with these changes. These results suggest that differences in the coagulation properties of milks containing either κ-casein A or κ-casein B are more likely to be related to factors, such as micelle size or charge, than to structural differences arising from altered backbone conformation of the macropeptide segments of the κ-caseins.

Milk and yogurt constitute a major source of dietary protein. The nutritive value of dietary proteins is linked to subsequent postprandial amino acid availability in the portal blood (Rérat, 1988). Portal absorption of nutrients cannot be studied in humans, but pigs provide a valid model for studying protein digestion in humans (Rowan et al. 1994).

Since stable isotopes are suitable to distinguish the exogenous from endogenous protein fraction in the intestinal lumen, intrinsic isotopic labelling of milk proteins has been considered a useful technique for nutritional investigations (Gaudichon et al. 1995; Gaudichon et al. 1999; Mahé et al. 1994). Recently, the use of 15N-labelled milk proteins were used to distinguish exogenous from endogenous N fractions in the human intestine after ingestion of 15N-milk or 15N-yogurt (Gaudichon et al. 1995). These authors pointed out that the jejunal flux of 15N was different for milk and yogurt. It is known that milk proteins and lactose undergo preliminary hydrolysis during lactic fermentation (Tamine & Deeth, 1980). It is also suggested that lactic fermentation enhances the nutritional value of milk proteins (Vass et al. 1984).

The effect of long-term intake of two fermented milk products on the development of hypertension was compared in young spontaneously hypertensive rats (SHR). The products contained tripeptides isoleucine-proline-proline (IPP) and valine-proline-proline (VPP), which have been shown to possess angiotensin converting enzyme (ACE) inhibitory activity. Six-week-old SHR were divided into four groups to receive orally ad libitum water, skim milk or two fermented milk products (fermented milk A or fermented milk B; the latter is commercially available in Japan with trade name Calpis®) for 14 weeks. The calculated intake of IPP was 0·4 mg/d and 0·2 mg/d in the groups receiving fermented milk A and B, respectively, whereas the corresponding amounts for VPP were 0·6 mg/d and 0·3 mg/d. Systolic blood pressure (SBP) was monitored weekly by tail-cuff method. The development of hypertension was significantly attenuated in both groups receiving fermented milk products, whereas skim milk did not affect blood pressure. The effect was detectable after 6 weeks of treatment. At the end of the experiment, the lowest blood pressure level was found in the group receiving fermented milk A: the SBP was 21 mm Hg lower than in the group receiving water and 10 mm Hg lower than in the group receiving fermented milk B. This difference could be explained by larger intake of ACE inhibitory tripeptides in the group receiving fermented milk A as compared with fermented milk B.

The effects of high intensity pulsed electric field (HIPEF) treatments at room or moderate temperature on water-soluble (thiamine, riboflavin, ascorbic acid) and fat-soluble vitamins (cholecalciferol and tocopherol) were evaluated and compared with conventional thermal treatments. Vitamin retention was determined in two different substrates, milk and simulated skim milk ultrafiltrate (SMUF). Samples were subjected to HIPEF treatments of up to 400 μs at field strengths from 18·3 to 27·1 kV/cm and to heat treatments of up to 60 min at temperatures from 50 to 90 °C. No changes in vitamin content were observed after HIPEF or thermal treatments except for ascorbic acid. Milk retained more ascorbic acid after a 400 μs-treatment at 22·6 kV/cm (93·4%) than after low (63 °C-30 min; 49·7% retained) or high (75 °C-15 s; 86·7% retained) heat pasteurisation treatments. Retention of ascorbic acid fitted a first-order kinetic model for both HIPEF and thermal processes. First-order constant values varied from 1·8×10−4 to 1·27×10−3 μs−1 for the HIPEF treatments (18·3–27·1 kV/cm) and, for thermal processing ranged from 5×10−3 to 8×10−2 min−1 (50–90 °C). No significant differences were found between the results obtained after applying HIPEF treatments at room or moderate temperature. However, results depended on the treatment media. A beneficial effect of natural skim milk components, mainly proteins, was observed on the preservation of ascorbic acid, since skim milk retained more ascorbic acid than SMUF after HIPEF treatments.

The influence of carbonation on the evolution of lactose, galactose and glucose in fermented milks with added probiotic bacteria (Lactobacillus casei, Lactobacillus acidophilus and/or Bifidobacterium bifidum) was evaluated and related to β-galactosidase activity of starter strains. During incubation and first days of refrigeration, lactose hydrolysis resulting in the liberation of galactose and glucose occurred in CT (Streptococcus thermophilus/Lb. casei), AT (Str. thermophilus/Lb. acidophilus) and ABT fermented milks (Str. thermophilus/Lb. acidophilus/Bifid. bifidum). Levels of galactose were higher than those of glucose and could be related to the preferential consumption of glucose by actively growing bacteria. Through the incubation, lactose and monosaccharide levels were not affected by milk carbonation. However, during refrigerated storage the presence of this gas was associated with slightly lower content of lactose and higher levels of galactose and glucose in AT and ABT products but not in CT fermented milks. Through the refrigeration galactose was moderately utilised by Lb. acidophilus in AT products whereas the presence of Bifid. bifidum seems to prevent the consumption of this sugar in ABT fermented milks. Glucose remained constant, with minor variations in CT products but a continuous increase of this sugar occurred in carbonated AT and ABT fermented milks during storage. β-Galactosidase activity displayed by Str. thermophilus strains was similar at pH 6·5 (initial pH of non-carbonated samples) and pH 6·3 (initial pH of carbonated samples) whereas Lb. acidophilus LaA3 showed greater β-galactosidase activity at pH 6·3 than at higher pH values. Thus, the enhanced metabolic activity of Lb. acidophilus caused by the low initial pH of carbonated milk also promoted higher cellular β-galactosidase activity that could have released greater amounts of galactose and glucose from lactose in AT and ABT fermented milks through the refrigerated period. In CT fermented milks, similar β-galactosidase activity levels of Str. thermophilus at pH 6·5 and 6·3 together with the absence of β-galactosidase activity in Lb. casei could explain the lack of differences on glucose and galactose content between carbonated and non-carbonated samples.

In the present work, 67 strains of Lactobacillus helveticus isolated from whey starter cultures and cheeses were identified and grouped by genotypic and phenotypic methods. Strains were identified by sugar fermentation pattern, by cell-wall protein profile, and by probe hybridisation. Phenotypic diversity was evaluated by a chemometric model taking into account biochemical characteristics (i.e. acidifying and peptidase activities) of technological interest. Genotypic diversity was evidenced by RAPD-PCR, which provided strain-specific patterns and revealed the occurrence of different strains. The RAPD-PCR profiles were clustered according to their similarities; the groups obtained, together with the cell-wall protein profiling and the chemometric information, could be sometimes correlated with the type of cheese and/or dairy niches used as sources of strains. A computerised analysis of genotypic and phenotypic information could be successfully applied for rapid and reliable differentiation and characterisation of Lb. helveticus isolates occurring in different dairy products.

Use of electromyography (EMG) to monitor mastication is a relatively new concept in assessing food physical and sensory properties. Although expert assessment of cheese characteristics is widely used, the effect of training in sensory analysis on mastication patterns, as assessed using EMG is not well known. Nine samples of the same Mahon cheese (60 days ripening) were given to 24 subjects (8 experts, 16 untrained) and EMG recorded for each chewing sequence. Three samples were tested in a single session by each subject, and three sessions carried out on different days. EMG was recorded from four masticatory muscles for each subject. From EMG records the following was extracted: number of chews, chewing time, mean and maximum voltage of EMG bursts (i.e. chews) across chewing sequence, chewing work and chewing rate. No gender bias was found for the EMG parameters considered, therefore, as regards gender, each group was considered to be homogeneous. Variability within-subjects across samples was greater for experts than untrained subjects. Significant differences in chewing time, chewing work and chewing rate were found between the expert and untrained groups. Data analysis of the three sessions showed an influence of cognitive constructs, mediating states, on the chewing process. The experts were found to be goal driven as to their mastication process. Experts showed no significant differences between sessions, untrained subjects were found to vary their EMG output in successive sessions for number of chews, chewing time, mean voltage, and chewing work.

Desi ghee (DG) was prepared from fermented cream followed by heat clarification (desi method) in the laboratory and butter oil (BO) was prepared from fresh butter by melting and centrifugation. Fresh samples of three brands of industrial ghee (IG-1, IG-2, IG-3) were collected from the local market. Volatile compounds of desi and industrial ghee and butter oil were isolated and concentrated using direct injection and cryofocussing techniques; separation and identification was by Gas Chromatography Mass Spectrometry (GCMS). A maximum of 36 compounds were detected in desi ghee whereas compounds detected in three industrial ghee samples varied from 22–29. The lowest number of compounds (16) was detected in butter oil. Of the identified compounds, maltol, 5-hydroxymethyl furfuraldehyde, dihydrodihydroxypyranone, 1,3-butanediol and 1-octanol were identified only in desi ghee volatiles. The concentration of acetic acid was found to be remarkably higher in desi ghee volatiles than in industrial ghee. Also the levels of identified fatty acids, methyl ketones, aldehydes, lactones and alcohols were high in desi ghee volatiles compared with industrial ghee and butter oil. In total, 62 compounds were detected, which included 6 aldehydes, 12 ketones, 8 each of fatty acids, alcohols and lactones, 4 each of esters and hydrocarbons or other compounds, and 12 compounds remained unidentified.