Experiments were designed to investigate the suitability of a combination of a short manual teat stimulation with a short latency period before teat cup attachment to induce and maintain oxytocin release and milk ejection without interruption. In Experiment 1, seven dairy cows in mid lactation were manually pre-stimulated for 15, 30 or 45 s, followed by either 30 s or 45 s of latency period. It was shown that all treatments induced a similar release of oxytocin without interruption until the end of milking. In particular, the latency period of up to 45 s did not cause a transient decrease of oxytocin concentration. In Experiment 2, milking characteristics were recorded in seven cows each in early, mid, and late lactation, respectively. Because the course of milk ejection depends mainly on the degree of udder filling, individual milkings were classified based on the actual degree of udder filling which differs between lactational stages but also between morning and evening milkings. All animals underwent twelve different udder preparation treatments, i.e. 15, 30, or 45 s of pre-stimulation followed by latency periods of 0, 30, 45, or 60 s. Milking characteristics were recorded. Total milk yield, main milking time and average milk flow rate did not differ between treatments if the degree of udder filling at the start of milking was >40% of the maximum storage capacity. However, if the udder filling was <40%, main milking time was decreased with the duration of a latency period up to 45 s, independent of duration of pre-stimulation. Average milk flow at an udder filling of <40% was highest after a pre-stimulation of 45 s followed by a latency period of another 45 s. In contrast, average milk flow reached its lowest values at a pre-stimulation of 15 s without additional latency period. However, average milk flow after a 15-s pre-stimulation increased with increasing latency period. In conclusion, a very short pre-stimulation when followed by a latency period up to 45 s before teat cup attachment remains a suitable alternative for continuous stimulation to induce milk ejection.

Ovine and caprine milks showed a marked stability maximum at ~ pH7 in their heat-stability/pH curves, but became very unstable at higher pH values. A low level of κ-casein appears to be responsible for this low stability of ovine milk; β-lactoglobulin offsets the stabilizing influence of κ-casein at lower pH values. Removal of colloidal calcium phosphate from ovine or caprine milks had very little influence on their heat stabilities and it was necessary to reduce the concentration of soluble salts to a very low level before an effect was observed. At the pH of maximum stability ovine and caprine milks, although quite variable, had stabilities in the same range as bovine milks. Equine milk was very unstable to heat and the shape of the heat-stability/pH curve of most individual samples was similar to ovine and caprine milks.

The effect of intramammary infusion of Escherichia coli endotoxin upon mammary blood flow was studied in lactating goats and cows. Blood flow was recorded by means of an electromagnetic flow probe chronically implanted around one mammary artery.

Endotoxin mastitis was always accompanied by a significant increase in mammary blood flow, characterized by 2 conspicuous peaks. The flow returned to control values by the thirteenth hour after infusion. Other symptoms of acute mastitis were: fever, increased heart rate, swelling, heat and pain of the gland, increased chloride and total cell count in milk.

Monospecific rabbit antibodies were produced against bovine chymosin (rennin) and bovine pepsin A and used for quantitative measurements of these enzymes by rocket immunoelectrophoresis. Bovine pepsin B was identified and measured quantitatively in tandem crossed immunoelectrophoresis, using a polyspecific antibody preparation. Chromatographically purified chymosin B and bovine pepsins A and B were used as standards. The contents of chymosin and bovine pepsin A were also determined by 2 forms of DEAE chromatography, and by an assay utilizing the ratio of milk-clotting activity at pH 6·0 and 6·5. Results obtained by measurements on 3 bovine rennets (calf rennet, adult bovine rennet, and a mixture of such types) by the different methods of analysis were in good agreement. The advantages of the immunochemical methods are direct and simultaneous identification and quantitation of enzymes.

The objective of this study was to investigate whether there is a difference in chemical composition and particularly in fatty acid (FA) profile, with emphasis on cis-9, trans-11 CLA, of milk obtained from conventional and organic dairy sheep and goats farms under the farming conditions practiced in Greece. Four dairy sheep and four dairy goat farms, representing common conventional production systems and another four dairy sheep and four dairy goat farms, organically certified, representing organic production and feeding systems were selected from all over Greece. One hundred and sixty two individual milk samples were collected from those farms in January–February 2009, about three months after parturition. The milk samples were analyzed for their main chemical constituents and their FA profile. The results showed that the production system affected milk chemical composition: in particular fat content was lower in the organic sheep and goats milk compared with the corresponding conventional. Milk from organic sheep had higher content in MUFA, PUFA, α-LNA, cis-9, trans-11 CLA, and ω-3 FA, whereas in milk from organic goats α-LNA and ω-3 FA content was higher than that in conventional one. These differences are, mainly, attributed to different feeding practices used by the two production systems. The results of this study show that the organic milk produced under the farming conditions practiced in Greece has higher nutritional value, due to its FA profile, compared with the respective conventional milk.

Citrate stimulated growth, totally induced citrate lyase, partly induced acetolactate synthase activity and partly repressed both diacetyl and acetoin reductases in Leuconostoc lactis NCW1. Similar results were obtained with 2 other leuconostocs and a heterofermentative lactobacillus. In 2 of the 3 leuconostocs tested, diacetyl reductase and acetoin reductase were NADPH specific, while in the 2 heterofermentative lactobacilli, they were NADH specific.

The current work studied four types of binary antagonist/pathogen bacterial culture system, in order to determine the effect of interaction between two strains of Lactobacillus plantarum and two food-borne pathogens, Listeria monocytogenes and Escherichia coli, in whole UHT milk at 37°C. To determine the type of interaction between the two bacterial populations in co-cultures and to evaluate the antagonistic activity of the lactic acid bacteria (LAB) on the pathogenic bacteria, the growth curves, the kinetic parameters, and the pH profiles of mono- and co-cultures were compared. The Lb. plantarum strains showed different bacteriocin-like inhibitory substance (BLIS) production, auto- and co-inducible. The antibacterial effect of neutralized supernatants of mono and co-cultures harvested at different times of incubation was assessed in order to establish the presence of bacteriocin-like inhibitory-substances (BLIS) and their possible relation to the growth inhibition of the pathogen. The LAB reduced the growth of Esch. coli and of List. monocytogenes by 4 and ∼5 log cycles, respectively and influenced other growth kinetic parameters, such as μmax and lag phase, in the different binary combinations. The growth of the LAB was not relevantly altered by simultaneous growth with the pathogenic strains showing an interaction of amensalism. The pattern of inhibition exerted by the LAB on the pathogens was different; Lb. plantarum LB279 inhibited the growth of List. monocytogenes more effectively than that of Esch. coli. The behaviour of Esch. coli in co-culture with Lb. plantarum WS4174 suggested the presence of metabolic crowding in the mechanism of growth suppression. This exploratory study showed the complexity and specific particularities of the inhibition phenomena between bacterial communities.

Analysis by gas chromatography–mass spectrometry of pasteurized milk with a fruity (pineapple-like) off-odour and a sour, rancid and soapy taste indicated the presence of concentrations at μg/ml levels of ethyl butanoate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, octanoic acid, decanoic acid and dodecanoic acid. The off-odour and taste were attributed to the presence of these compounds in the milk. Microbiological examination confirmed that the milk was also contaminated with a series of psychrotrophic bacteria including Yersinia intermedia, Pseudomonas putida and Rahnella aquatilis. Growth of isolates of these bacteria in UHT milk at 23 °C for 7 d showed that Yer. intermedia produced significant quantities of the C4–C12 alkanoic acids; Ps. putida produced only small quantities of these acids and Rah. aquatilis produced none. In addition, Yer. intermedia and Ps. putida also produced small but significant quantities of the corresponding ethyl esters. In milk inoculated with both Yer. intermedia and Ps. putida, the quantity of ethyl esters produced was greater than that found in cultures containing only one of the isolates. These studies indicated that Yer. intermedia was the principal source of the alkanoic acids in the tainted milk and that the major producer of the corresponding ethyl esters was Ps. putida. This is the first report that Yer. intermedia and Ps. putida can cause an off-odour or taste in dairy products.

Proteinase activity of Lactobacillus fermentum IFO3956 cells was higher when they were grown on milk-based media than on 10% reconstituted skim milk. The lowest protease activity was observed when cells were grown on milk-free media. The extraction of milk-induced cell-bound proteases from Lb. fermentum IFO3956 was most efficient using 1% Tween 80 while the use of 1% SDS inhibited all proteolytic activity. Two bands of ∼35 and >100 kDa were observed by zymogram, indicating that proteolytic activity corresponded to the presence of at least two types of enzymes or two molecular forms of one enzyme. Mass spectrometry analyses of αS1-casein hydrolysates detected 24 peptides with sizes ranging from 5 to 36 amino acids, including 9 phosphorylated peptides, resulting from the fermentation of Lb. fermentum IFO3956 of αS1-casein. Most of the identified peptides originated from the N-terminal portion of αS1-casein. The studied bacterial strain could hydrolyze αS1-casein in many sites including the epitopes triggering the allergic reactions against αS1-casein e.g. at the positions 23, 30, 41, 71, 91, 98, 126, 179. After hydrolysis of αS1-casein with Lb. fermentum IFO3956 the recognition and the binding of this casein to IgE from the pooled sera of 18 patients with cow's milk allergy was significantly reduced.

The transfer of ingested α-linolenic acid (ALA) and linoleic acid (LA) determines the nutritional quality of milk, but the factors determining this transfer are unclear. The present experiment investigated the influence of roughage to concentrate proportions and the effect of concentrate types on milk fat composition. Respectively, six lactating dairy cows were fed one of three isoenergetic (5·4±0·05 MJ net energy for lactation/kg dry matter; DM) and isonitrogenous (215±3·5 g crude protein/kg DM) diets, consisting of ryegrass hay only (33 g fatty acids/kg DM; ALA-rich, no concentrate), maize (straw, whole maize pellets and gluten; 36 g fatty acids/kg DM; LA-rich; 560 g concentrate/kg DM), or barley (straw and grain plus soybean meal; 19 g fatty acids/kg DM; LA-rich; 540 g concentrate/kg DM). The fatty acid composition of feeds and resulting milk fat were determined by gas chromatography. The ALA concentration in milk fat was highest (P<0·001) with the hay-diet, but the proportionate transfer of ALA from diet to milk was lower (P<0·001) than with the maize- or barley-diets. The LA concentration in milk fat was highest with the maize-diet (P<0·05, compared with hay) but relative transfer rate was lower (P=0·01). The transfer rates of ALA and LA were reciprocal to the intake of individual fatty acids which thus contributed more to milk fat composition than did roughage to concentrate proportions. The amount of trans-11 18:1 in milk fat was lowest with the barley-diet (P<0·001) and depended on the sum of ALA and LA consumed. The milk fat concentration of cis-9, trans-11 18:2 (rumenic acid) was more effectively promoted by increasing dietary LA (maize) than ALA (hay). Amounts of 18:0 secreted in milk were four (maize) to seven (hay) times higher than the amounts ingested. This was suggestive of a partial inhibition of biohydrogenation in the maize-diet, possibly caused by the high dietary LA level.

To establish whether fishy off-flavour observed in cows' milk in Sweden is due to abnormal concentrations of trimethylamine (TMA) in milk, 15 milk samples from 13 cows of the Swedish Red and White breed (SRB) were analysed for this compound using dynamic headspace (DHS) gas chromatography (GC). A mass selective detector (MSD) was used for the qualitative and a flame ionization detector (FID) for the quantitative analyses. Further confirmation was obtained using a solid phase microextraction (SPME) followed by GC-olfactometry (GC-O) and GC-NPD (nitrogen and phosphorus detector). Samples were also evaluated by a two-person organoleptic panel for the presence of fishy off-flavour. Results showed that the seven milk samples with a fish-like taint contained >1 mg TMA/kg milk, whereas, with a single exception, the seven samples that were judged to be normal did not contain this compound. Furthermore, there were indications of a dose-dependent relationship between TMA concentration and fishy off-flavour score.

The aim of this study was to determine the influence of the somatic cell content (SCC) of milk on Parmigiano-Reggiano cheese yield, produced in commercial cheese factories under field conditions. The study was carried out following the production of 56 batches of Parmigiano-Reggiano in 13 commercial cheese factories by processing milk collected from Italian Friesian cattle herds. The vat-milk (V-milk) used for making each cheese batch was obtained by mixing evening milk (partially skimmed following spontaneous separation of fat overnight, natural creaming) and morning milk. The batches of cheese produced were divided into 5 classes according to the SCC value of the evening milk determined prior to natural creaming (class 1, from 0 to 200 000; 2, 201 000–300 000; 3, 301 000–400 000; 4, 401 000–500 000; 5, over 501 000 cells/ml). The cheese yield was calculated as the amount of 24-h cheese, expressed in kilograms, obtained from 100 kg of V-milk (24 h ACY). The values of fat, crude protein, true protein, casein and 24 h ACY of V-milk were negatively correlated with the somatic cell score (SCS) of the evening milk. Conversely, a positive correlation was observed between chloride and SCS. Fat, protein fractions (crude protein, casein and whey proteins), P and titratable acidity of V-milk were positively correlated with its 24 h ACY, while chloride, pH and SCS showed a negative correlation. A significant drop in 24 h ACY was observed in classes 3, 4 and 5, therefore when the SCC of the evening milk exceeded 300 000 cells/ml. Finally a lower recovery of milk fat in cheese was observed as SCC of evening milk increase.

In order to predict the oozing susceptibility of heat-induced milk protein gels such as processed cheeses during storage, ultracentrifugation forces are required to accelerate the expulsion of water from the gel structure. Two predictive methods have been tested. Direct ultracentrifugation of processed cheese was used to study effects of centrifugal compression on the water-holding ability. The water release with optimal parameters (time, temperature and ultracentrifugation force) from a freshly manufactured processed cheese was correlated with visual assessment of oozing after 6 months storage. A second method was based on the susceptibility of gelled proteins to resist the dissociating action of solutions of chemical agents such as SDS, urea, EDTA and 2-mercaptoethanol. Most of the protein sedimented by ultracentrifugation in the presence of SDS represented that still in complexed form. This correlated with optimal water binding by the processed cheese. Response-surface methods, used to optimize the ultracentrifugation variables and the composition of the dissociation solutions, showed that the dissociation test was best with a dispersion of processed cheese in SDS solution (10 g/l) at a ratio of 1:6 (w/v) at 20°C, ultracentrifuged at 86000 g and 20°C for 25 min. Analysis of processed cheese cooked at 115°C and then dissociated in different solutions showed various protein–protein interactions in the gel network, whereas hydrophobic interactions were the most important stabilizers of the protein matrix of cheese cooked at higher temperature.

The extent and level of proteolysis were evaluated in Cabrales and five other European blue vein cheese varieties, Bleu de Bresse, Danablu, Edelpilzkäse, Gorgonzola and Roquefort, by analysing their different N-containing constituents and determining the degree of casein hydrolysis. The electrophoretic patterns of the caseins in the cheeses confirmed their strong proteolysis, components featuring the lowest electrophoretic mobility offering the greatest resistance to subsequent hydrolysis. Component αs1 — I was detected in all varieties studied with the exception of Roquefort cheese.

The changes in milk fat globules and fat globule surface proteins during concentration of whole milk using a pilot-scale multiple-effect evaporator were examined. The effects of heat treatment of milk at 95 °C for 20 s, prior to evaporation, on fat globule size and the milk fat globule membrane (MFGM) proteins were also determined. In both non-preheated and preheated whole milk, the size of milk fat globules decreased while the amount of total surface proteins at the fat globules increased as the milk passed through each effect of the evaporator. In non-preheated samples, the amount of caseins at the surface of fat globules increased markedly during evaporation with a relatively small increase in whey proteins. In preheated samples, both caseins and whey proteins were observed at the surface of fat globules and the amounts of these proteins increased during subsequent steps of evaporation. The major original MFGM proteins, xanthine oxidase, butyrophilin, PAS 6 and PAS 7, did not change during evaporation, however, PAS 6 and PAS 7 decreased during preheating. These results indicate that the proteins from the skim milk were adsorbed onto the fat globule surface when the milk fat globules were disrupted during evaporation.

The leucocytes in sheep colostrum, milk and involution secretion were studied using the electron microscope. The predominant cell type in colostrum was the polymorphonuclear leucocyte (PMNL) (41–84%), followed by the macrophage (8–49%), and the lymphocyte (6–11 %). Plasma cells were present in low numbers (1–2%) while no secretory epithelial cells were observed. In mid-lactation the cell components changed so that the macrophage was the predominant cell (83–86%) followed by the lymphocyte (10–17%). In early involution secretion, PMNL reappeared but declined in secretion obtained 21 d after weaning, when the macrophage was again the predominant cell. Associated with the whole cells were membranous extracellular materials and ‘sunburst’ fragments of the secretory epithelial cells. These were phagocytosed by both PMNL and macrophages, which also engulfed fat droplets from the milk. The lymphocytes were examined for plasma membrane markers and the T-cell percentage (E+ rosettes) fluctuated within wide limits, (0–80 %). However, there was no discernible trend associated with stage of lactation. The B-cell markers (C1 and Fcγ) also fluctuated widely and it was evident that contaminating monocytes reduced the accuracy of the count. However, it appeared that more B-cells were present in milk than in blood.

Sedimentation Field Flow Fractionation (SdFFF) was combined with Photon Correlation Spectroscopy (PCS), to characterize changes in the structure of the colloidal particles of reconstituted skim milk of diameter >50 nm (aggregates of casein and calcium phosphate known as casein micelles) with the changes in partitioning (with the addition of salt) of calcium (Ca), inorganic phosphate (Pi) and casein between the serum and colloidal phases of the milk. The number weighted particle size distributions are determined. These are well represented by a log-normal distribution. Methods are presented for estimating the relative contributions of scattering and absorbance to the SdFFF detector signal and for taking both into account when analysing SdFFF data. The values found for the effective density of the casein micelles were in good agreement with the literature and ranged from (1·06–1·08 g cm−3) according to the composition of micelles. The changes in the scattering intensity as determined by PCS correlated with the changes in the particle composition. Although the concentrations of colloidal calcium phosphate (CCP) (1·1–3·5 g/kg milk) and micellar casein (18·1–27·2 g/kg milk) varied considerably only small changes in the size distribution of particles >50 nm diameter were observed except for milk to which 30 mmol Pi+10 mmol Ca/kg milk had been added where the particle size distribution shows a swelling of the particles consistent with a lower than expected value for the particle density. These observations suggest that the micelles have the ability to both lose (depleted micelles) and accommodate (enriched micelles) more casein, calcium and inorganic phosphate in their interior, thus confirming the model of the micelles which postulates an open structure allowing freedom of movement of casein and small ions.

Subclinical mastitis (SM) is one of the most important diseases affecting dairy ewes worldwide, with negative impact on the animal health, farm income and public health. Animals with SM often remain untreated because the disease may not be revealed. Increase in somatic cell count (SCC) and positive bacteriology for mastitis pathogens in milk samples are indicative of SM but the evidence of only one of these alterations must suggest an uncertain SM (UM). UM is defined when positive bacteriological examination (Latent-SM) or SCC>500 000 cells/ml (non-specific-SM) are detected in milk. Nevertheless, SCC and bacteriological examination are expensive, time consuming and are not yet in use at the farm level in dairy ewes. Recently, a sensitive acute phase protein, amyloid A, displaying multiple isoforms in plasma and different body fluids including mammary secretion (milk amyloid A-MAA), has been investigated as a marker of mastitis in cows and, in a few studies, in sheep. The aim of this trial was to compare the concentration of MAA of single udder-halves in ewes with healthy udder-halves (HU-control group) and naturally occurring subclinical mastitis, both confirmed (SM group) and uncertain (UM groups: Latent-SM and non-specific-SM), for monitoring udder health. The reliability of a specific ELISA kit for the measurement of MAA was also tested. During a 3-month trial period, 153 udder halves were assigned to the experimental groups based on their health status: 25 with SM, 40 with UM (11 with latent-SM and 29 with non-specific-SM) and 88 HU. SCC and bacteriological analysis were performed to establish the control and subclinical mastitis groups. MAA concentrations in milk samples were measured using a specific commercially milk ELISA kit. The data were submitted to statistical analysis. Significant (P<0·05) differences among the groups SM, non-specific-SM and HU were detected with the SM having the highest level and HU the lowest. MAA concentration is affected by the udder health status and is a useful indicator of subclinical mastitis and increased SCC in sheep.

Variations in chemical composition and in some physico-chemical properties of milk from a herd of lactating Alpine goats imported into Greece were determined from week 8 after parturition until the end of lactation (35 weeks, 35 samples). The average composition (g/100 g) of the milk was: fat 3·44, protein 3·35, casein 2·46, lactose 4·30, ash 0·79, total solids 11·76 and solids-not-fat 8·32. The average pH was 6·57 and titratable acidity was 16·76 (°D). The averaged density was 1·030. The mean mineral contents (mg/100 g) of the milk were: Na 53·12, K 157·67, Ca 140·06, P 110·00, Mg 13·59, citrate 106·97. All constituents studied as well as pH and density were significantly (P < 0·05) dependent on the stage of lactation. Titratable acidity was not significantly affected by stages of lactation. There were significant (P < 0·001) differences in Na/K and Ca/P ratios in the milk between lactation stages. Trends with lactation for each of the components and physico-chemical properties determined are reported and correlations between constituents in the milk are given.