The main requirement for milk processed in most cheese typologies is its rennet coagulation ability. Despite the increasing number of studies, the causes for abnormal coagulation of milk are not fully understood. The aim of this study was to ascertain relationships between milk characteristics and its rennet coagulation ability, focusing on the influence of calcium (Ca) and phosphorus (P). Ca and P are essential constituents of the micelles. Micellar P can be present as part of colloidal calcium phosphate (inorganic-P) or covalently bound to caseins as phosphate groups (casein-P). Eighty one herd milk samples (SCC<400 000 cell/ml) were classified as Optimal (8), Suboptimal (39) Poor (29) and Non-coagulating milk (5), according to their rennet coagulation parameters as assessed by lactodynamographic test. Samples were analysed for their chemical composition (basic composition, protein fractions, minerals and salt equilibria), physicochemical parameters (pH and titratable acidity) and rheological properties. Optimal milk was characterised by the highest contents of major constituents, protein fractions and minerals, lowest content of chloride and highest values of titratable acidity. Non-coagulating milk was characterised by the highest values of pH and the lowest of titratable acidity. At micellar level, Optimal milk showed the highest values of colloidal Ca, casein-P and colloidal Mg (g/100 g casein), while Non-coagulating milk showed the lowest values. Interestingly, there was no statistical difference regarding the content of colloidal inorganic-P (g/100 g casein) between Optimal and Non-coagulating milks. Overall, high mineralisation of the micelle (expressed as g inorganic-P/100 g casein) positively affect its rennetability. However, excessive mineralisation could lead to a reduction of the phosphate groups (g casein-P/100 g casein) available for curd formation.

Cell-free supernatant from Leuconostoc citreum MB1 revealed specific antilisterial activity. Preliminary studies demonstrated the proteinaceous, heat-stable, bacteriocin-like trait of the antimicrobial components present in the supernatant. Determination of the genes encoding bacteriocins by PCR and DNA sequencing led to amplification products highly homologous with leucocin A (found in diverse Leuconostoc species) and UviB (found in Leuc. citreum KM20) sequences. Additionally, antimicrobial activity of cell-free supernatant from Leuc. citreum MB1 was revealed by an inhibition halo of the SDS-PAGE gel subjected to a direct detection using Listeria monocytogenes as indicator strain. Different assays were carried out to assess the capacity of Leuc.citreum MB1 to control List. monocytogenes growth: (i) inactivation kinetics of the pathogen by antilisterial compounds present in concentrated cell-free supernatant from Leuc. citreum MB1, (ii) evaluation of optimal Leuc. citreum MB1 initial concentration to obtain maximum List. monocytogenes ATCC 15313 inhibition, and (iii) biocontrol of List. monocytogenes ATCC 15313 with Leuc. citreum MB1 during growth in milk at refrigeration temperature. According to our results, it is unquestionable that at least one bacteriocin is active in Leuc. citreum MB1, since important antilisterial activity was verified either in its cell-free supernatant or in co-culture experiments. Co-culture tests showed that ∼107 CFU/ml Leuc. citreum MB1 was the optimal initial concentration to obtain maximum pathogen inhibition. Moreover, Leuc. citreum MB1 was able to delay List. monocytogenes growth at refrigerated temperature.

Reconstituted skim milks (10 % w/w total solids, pH 6·7–8·0) were ultrasonicated (20, 400 or 1600 kHz at a specific energy input of 286 kJ/kg) at a bulk milk temperature of <30 °C. Application of ultrasound to milk at different pH altered the assembly of the casein micelle in milk, with greater effects at higher pH and lower frequency. Low frequency ultrasound caused greater disruption of casein micelles causing release of protein from the micellar to the serum phase than high frequency. The released protein re-associated to form aggregates of smaller size but with surface charge similar to the casein micelles in the original milk. Ultrasound may be used as a physical intervention to alter the size of the micelles and the partitioning of caseins between the micellar and serum phases in milk. The altered protein equilibria induced by ultrasound treatment may have potential for the development of milk with novel functionality.

This study investigated the effects of different strategies for feeding supplements to grazing dairy cows on the proportions of fatty acids in milk. Two hundred and sixteen cows were fed supplementary grain and forage according to one of 3 different strategies; (1) Control: cows grazed perennial ryegrass pasture (14 kg dry matter/d) supplemented with milled barley grain fed in the milking parlour and pasture silage offered in the paddock; (2) Partial mixed ration 1 (PMR1): same pasture allotment and supplement as Control strategy, but the supplements presented as a mixed ration after each milking in feedpad, and; (3) Partial mixed ration 2 (PMR2): same pasture allotment, supplemented with a mixed ration of milled barley grain, alfalfa hay, corn silage and crushed corn grain fed in a feedpad. Within each strategy, cows were assigned to receive either 6, 8, 10 or 12 kg dry matter supplement/cow per d. Milk fatty acid proportions from cows fed Control and PMR1 strategies were similar and different from those fed PMR2, particularly at 10 to 12 kg dry matter supplement/cow per d. The reduction in milk fat yield and concentration in cows fed high amounts of supplement as Control and PMR1 was coincident with 4 × increase in 10t-18:1 proportion. The composition of the partial mixed ration (PMR) and the amount offered affected milk fatty acid proportions and milk fat content, however, the method of supplementation did not.

Bacteriogenic Enterococcus faecium HZ was identified by using biochemical (Strep-API 20, API-50 CHL, fatty acid profile) and 16S rRNA analysis (99·99 %). Ent. faecium HZ was sensitive to clinically important antibiotics such as vancomycin, and did not have gelatinase and haemolysis activities. Enterocin HZ, a bacteriocin from Ent. faecium HZ, was sensitive to papain and tyripsin, but resistant to pepsin, lipase, catalase, α-amylase, organic solvents, detergents, ß-mercaptoethanol, and heat treatment (90 °C/30 min). It was biologically active at pH 2·0–9·0 and synthesised at the highest level in MRS or M17 broth at 32 or 37 °C with an inoculum amount of 0·1–0·5 % and an initial pH of 6·0–7·0. Enterocin HZ production reached maximum level at middle and late logarithmic phase and its molecular weight was ∼4·5 kDa. It was active against some Gram-positive foodborne bacteria. Ent. faecium HZ or its bacteriocin enterocin HZ is a good candidate to be studied as a food biopreservative since enterocin HZ showed strong bactericidal activity against Listeria monocytogenes in UHT milk and also Ent. faecium HZ grew very well in milk and produced enterocin HZ at maximum level.

In order to evaluate milking ability in dromedary camels, 124 milk flow curves were registered during morning milking of 20 dairy Maghrebi dromedary camels. Animals were in lactations 1–8, were 6–19 years old and were 4–15 months of their current lactation. Milk flow curves were recorded using an electronic milk flow meter (Lactocorder®). Milk flow curves were classified in three typical patterns: type 1 represents curves with one high and short peak of milk flow; type 2 represents curves with a moderate mean milk flow rate during a large plateau phase; and type 3 represents curves with lower mean milk flow rate and a relatively longer milking duration. The ratio of the different milk flow patterns in the population evaluated was 40:38:22% for types 1, 2 and 3, respectively. The highest milk yield per milking, average and peak milk flow were observed in camels with type 1 curves (4·24 kg, 1·49 and 3·54 kg/min, respectively) followed by type 2 animals (3·30 kg, 1·12 and 2·12 kg/min, respectively) and lastly type 3 curves (2·34 kg, 0·65 and 1·23 kg/min, respectively). This study confirmed that a major proportion of dromedary camels have a suitable machine milking ability. Nevertheless, our results suggest that pre-stimulation and improving the milking process may improve milking efficiency and guarantee a more complete and rapid emptying of the udder.

This experiment was conducted to determine the effects of diets supplemented with cottonseed oil, Acacia mearnsii-condensed tannin extract, and a combination of both on composition of bovine milk. Treatment diets included addition of cottonseed oil (800 g/d; CSO), condensed tannin from Acacia mearnsii (400 g/d; TAN) or a combination of cottonseed oil (800 g/d) and condensed tannin (400 g/d; CPT) with a diet consisting of 6·0 kg dry matter (DM) of concentrates and alfalfa hay ad libitum, which also served as the control diet (CON). Relative to the CON diet, feeding CSO and CPT diets had a minor impact on feed intake and yield of lactose in milk. These diets increased yields of milk and protein in milk. In contrast to the TAN diet, the CSO and CPT diets significantly decreased milk fat concentration and altered milk fatty acid composition by decreasing the proportion of saturated fatty acids but increasing proportions of monounsaturated and polyunsaturated fatty acids. The CPT diet had a similar effect to the CSO diet in modifying fatty acid profile. Overall, reduction in milk fat concentration and changes in milk fatty acid profile were probably due to supplementation of linoleic acid-rich cottonseed oil. The TAN diet had no effect on feed intake, milk yield and milk protein concentration. However, a reduction in the yields of protein and lactose occurred when cows were fed this diet. Supplemented tannin had no significant effect on fat concentration and changes in fatty acid profile in milk. All supplemented diets did not affect protein concentration or composition, nitrogen concentration, or casein to total protein ratio of the resulting milk.

In this study we identified Lactococcus lactis subsp. lactis, Lc. lactis subsp. lactis biovar diacetylactis, Kluyveromices lactis and Saccharomyces cerevisiae as the dominant microorganisms of traditional Moroccan acid-alcoholic fermented milk named Lben. The low pH (3·8±0·3), lactose (16·8±3·4 mg/l) and lactic acid (8·16±0·6 mg/l) content indicated that a strong fermentation occurred in the traditional product which was also characterised by the substantial presence of ethanol and typical volatile carbonyl compounds (i.e., acetoin, diacetyl and acetaldehyde). Microbiological analyses of experimental Lben manufactured with selected strains (isolated from the traditional product) of Lc. lactis subsp. lactis and Lc. lactis subsp. lactis biovar. diacetylactis alone (batch A) and in combination with enzymatic extract of a K. lactis strain (batch B) indicated a good effectiveness of the starters employed (∼1010 CFU/g of lactococci after 8 h of incubation) and a significant effect of the yeast enzyme extract on lactococci viability. Despite slight changes in the physicochemical characteristics of the two Lben during the 15 d storage period, volatile compounds (i.e. ethanol, acetaldehyde, diacetyl and acetoin) were consistently higher in batch B. Moreover, sensorial analysis performed after 15 d of storage, highlighted higher odour and flavour intensity, vegetable odour and viscosity in batch B while batch A displayed higher astringency.

Probiotic bacteria have been used in human nutrition for centuries and are now attracting more attention. In order to examine the immunological aspects of probiotic consumption, Lactobacillus rhamnosus LA68 was orally administrated using gavage to healthy C57BL/6 mice. After one month splenocytes were isolated, and analysed by flow cytometry. The magnitude of splenocyte proliferation upon stimulation with lipopolysaccharide and peptidoglycan and cytokine levels (IFN-γ, IL-6, IL-10 and IL-17) was assessed. Cytokine levels in the serum were also analysed. Oral application of strain LA68 leads to a significant decrease of CD3+, CD25+ and CD19+ cells, and an increase of CD11b+ and CD16/CD32+ positive cell populations in the mouse spleen. Increased sensitivity to stimulation through proliferation and IL-6 secretion was detected. Increased serum IFN-γ and decreased IL-10 levels were found. Our results show increased responsiveness of splenocytes, activation of the Th1 type of immune response, and a shift of leucocyte populations towards monocyte/granulocyte populations.

The objectives of this study were to compare the impact of different coagulase-negative species (CNS) on udder health measured in terms of individual quarter milk somatic cell count (SCC) and duration of intramammary infection, and to get some insight into most likely routes of infection for different CNS species. This longitudinal observational study was performed on four farms that were sampled at 4-week intervals for a total of 12 visits each. Quarters infected with CNS were followed through time with milk samples being submitted for bacteriological culture and SCC determination. PCR amplification of the internal transcribed spacer region and sequencing of the sodA and rpoB genes were used for species allocation. Pulsed-field gel electrophoresis (PFGE) was performed to assess strain identity. The percentage of quarters affected per farm varied between 6 and 35%, with the most frequently isolated CNS species being Staphylococcus epidermidis, followed by Staph. simulans, Staph. chromogenes and Staph. haemolyticus. It was possible to follow 111 intramammary infections due to CNS through time. Duration of infection had a mean of 188 d and was not significantly different between CNS species. Geometric mean quarter SCC overall was 132 000 cells/ml and was also not significantly different between CNS species. Despite the possibility of a different epidemiology of infection, the impact in terms of udder health seems to be similar for different CNS species.

The aim of this work was to study the effect on teat wall thickness and canal length in sheep and goats of overmilking for 2 min (OM+2) and of milking with used (AL; +3000 milkings) and twisted (TL; 45°) liners in sheep and goats, as well as the effect of milking goats with liners designed for sheep (SL, shorter length and diameter than liners for goats). To this end, we performed four experiments in goats and three in sheep, in a Latin square design with two experimental periods. During the experimental period 4 controls were carried out, performing ultrasound scans before and immediately after milking to determine the teat wall thickness (TWT), teat wall area (TWA), teat end area (TEWA) and teat canal length (TCL). OM+2 caused a significant increase in TWT, TWA, TEWA and TCL in goats and in TWA, TEWA and TCL in sheep. Liner features had a strong influence on the variables studied; aged liners caused significant changes in TWT and TCL in goats and in TWT in sheep; twisted liners produced a significant effect on the increase of TWT and TCL in goats, without reaching significance level in sheep; and milking goats with sheep liners led to a significant increase in TWT, TWA, TEWA and TCL. In practice, it is therefore important to avoid overmilking and the use of worn-out liners. It is also necessary to use liners designed for the morphological features of each species, taking special care to carry out periodic liner positioning revisions to ensure the benefits of pulsation on the teat end. Finally, it would be necessary to carry out long-term experiments to study whether the increase in thickness observed in some experiments is sufficient to affect milking efficiency and mammary gland health status.

The impact of mastitis on milk value per litre independent of the effect of mastitis on milk volume, was quantified for Ireland using a meta-analysis and a processing sector model. Changes in raw milk composition, cheese processing and composition associated with increased bulk milk somatic cell count (BMSCC) were incorporated into the model. Processing costs and market values were representative of current industry values. It was assumed that as BMSCC increased (i) milk fat and milk protein increased and milk lactose decreased, (ii) fat and protein recoveries decreased, (iii) cheese protein decreased and cheese moisture increased. Five BMSCC categories were examined from ⩽100 000 to >400 000 cells/ml. The analysis showed that as BMSCC increased the production quantities reduced. An increase in BMSCC from 100 000 to >400 000 cells/ml saw a reduction in net revenue of 3·2% per annum (€51·3 million) which corresponded to a reduction in the value of raw milk of €0·0096 cents/l.

This work applies culture-independent methods for the characterization of fungal populations (yeasts and moulds) naturally occurring in Sardinian ewe's milk sampled in the Italian areas with the largest dairy production (Sardinia and Lazio regions). Sequences of the D1/D2 variable domains at the 5′ end of the 26S rRNA gene were obtained by amplification of DNA directly isolated from milk, and this allowed identification of a total of 6 genera and 15 species of fungi. Among the 6 identified genera Geotrichum spp., Candida spp., Phaeosphaeriopsis spp., Pestalotiopsis spp. and Cladosporium spp. belong to the phylum of Ascomycota, while Cryptococcus spp. is part of the phylum of Basidiomycota. In particular, two genera (Pestalotiopsis and Phaeosphaeriopsis) and two species (Plectosphaerella cucumerina and Pryceomyces carsonii) have never been reported in dairy ecosystems before. Results provide evidence that several moulds and yeasts, previously described only in ovine cheeses, are transferred directly from raw milk. The knowledge of fungal consortia inhabiting sheep raw milk is a particularly relevant issue because several species are directly involved in cheese making and ripening, determining the typical aroma. On the other hand, spoilage yeasts and moulds are involved in anomalous fermentation of cheese and may be responsible for considerable economic losses and serious risks for consumers' health.

The aggregation of proteins after heating of calcium-fortified milks has been an ongoing problem in the dairy industry. This undesirable effect restricts the manufacture of calcium rich dairy products. To overcome this problem, a completely new approach in controlling the heat stability of dairy protein solutions, developed in our lab, has been employed. In this approach, high intensity, low frequency ultrasound is applied for a very short duration after a pre-heating step at ⩾70 °C. The ultrasound breaks apart whey/whey and whey/casein aggregates through the process of acoustic cavitation. Protein aggregates do not reform on subsequent post-heating, thereby making the systems heat stable. In this paper, the acid gelation properties of ultrasonicated calcium-enriched skim milks have also been investigated. It is shown that ultrasonication alone does not change the gelation properties significantly whereas a sequence of preheating (72 °C/1 min) followed by ultrasonication leads to decreased gelation times, decreased gel syneresis and increased skim milk viscosity in comparison to heating alone. Overall, ultrasonication has the potential to provide calcium-fortified dairy products with increased heat stability. However, enhanced gelation properties can only be achieved when ultrasonication is completed in conjunction with heating.

This study investigated the antibacterial properties of plasmin, the plasmin hydrolysis of bovine κ-casein and the fractions (named κC1, κC2, κC3, κC4, and κC5) liberated from it using RP-HPLC. The target bacteria were Escherichia coli, Staphylococcus aureus (pathogenic), Lactobacillus casei and Lactobacillus acidophilus (probiotic). Three peptides (kC1, kC3, and kC4) were found to have antibacterial activity, with κC3 peptide being the most active. The plasmin digest of bovine κ-casein proved to be stronger than any of its fractions in terms of antibacterial potential. Measurement of the minimum inhibitory concentration (MIC) showed that Gram-positive bacteria are generally more sensitive to antibacterial activity than Gram-negative bacteria. The MIC of nisin, as a bacteriocin peptide, was also measured. The three antibacterial peptides were identified using LC-Mass. The molecular mass of kC1, kC3, and kC4 corresponded to the f(17–21), f(22–24), and f(1–3) of bovine κ-casein, respectively. It was also found that the positive charge and hydrophobicity of a peptide are not key factors in antibacterial activity. On the whole, the present study demonstrated that the plasmin digest of κ-casein has a high antibacterial potential and can be considered as a natural antibacterial agent in the food chain.

In this study, we developed mathematical models to describe the growth kinetics of Staphylococcus aureus on natural cheeses. A five-strain mixture of Staph. aureus was inoculated onto 15 g of Brie and Camembert cheeses at 4 log CFU/g. The samples were then stored at 4, 10, 15, 25, and 30 °C for 2–60 d, with a different storage time being used for each temperature. Total bacterial and Staph. aureus cells were enumerated on tryptic soy agar and mannitol salt agar, respectively. The Baranyi model was fitted to the growth data of Staph. aureus to calculate kinetic parameters such as the maximum growth rate in log CFU units (rmax; log CFU/g/h) and the lag phase duration (λ; h). The effects of temperature on the square root of rmax and on the natural logarithm of λ were modelled in the second stage (secondary model). Independent experimental data (observed data) were compared with prediction and the respective root mean square error compared with the RMSE of the fit on the original data, as a measure of model performance. The total growth of bacteria was observed at 10, 15, 25, and 30 °C on both cheeses. The rmax values increased with storage temperature (P<0·05), but a significant effect of storage temperature on λ values was only observed between 4 and 15 °C (P<0·05). The square root model and linear equation were found to be appropriate for description of the effect of storage temperature on growth kinetics (R2=0·894–0·983). Our results indicate that the models developed in this study should be useful for describing the growth kinetics of Staph. aureus on Brie and Camembert cheeses.