Prevention of the heat-induced aggregation of β-lactoglobulin (β-Lg) would improve the heat stability of whey proteins. The effects of lipoic acid (LA, or thioctic acid), in both its oxidised and reduced form (dihydrolipoic acid, DHLA), on heat-induced unfolding and aggregation of β-Lg were investigated. LA/DHLA was added to native β-Lg and the mixture was heated at 70, 75, 80 or 85 °C for up to 30 min at pH 6·8. The samples were analysed by Polyacrylamide Gel Electrophoresis (PAGE) and Size-exclusion HPLC (SE-HPLC). LA was not as effective as DHLA in reducing the formation of aggregates of heated β-Lg. Heating β-Lg with DHLA resulted in formation of more β-Lg monomers (due to dissociation of native dimers) and significantly less β-Lg aggregates, compared with heating β-Lg alone. The aggregates formed in the presence of DHLA were both covalently linked, via disulphide bonds, and non-covalently (hydrophobically) linked, but the amount of covalently linked aggregates was much less than when β-Lg was heated alone. The results suggest that DHLA was able to partially trap the reactive β-Lg monomer containing a free sulphydryl (−SH) group, by forming a ‘modified monomer’, and to prevent some sulphydryl−sulphydryl and sulphydryl−disulphide interactions that lead to the formation of covalently linked protein aggregates. The effects of DHLA were similar to those of N-ethylmaleimide (NEM) and dithio(bis)-p-nitrobenzoate (DTNB). However, the advantage of using DHLA over NEM and DTNB to lessen aggregation of β-Lg is that it is a food-grade compound which occurs naturally in milk.

Dairy cattle in the Alps are traditionally maintained on high altitude pastures during summer. In recent decades, however, many farmers prefer to maintain the cows always indoor with a hay-based diet. Many authors have shown that the forage type is able to modify the characteristics of milk and cheese. Recently the product specification of PDO Montasio allowed differentiation between mountain cheeses and other products. Aim of this trial is to study the effect of rearing system on the characteristics of milk and cheese produced in this context. One hundred and twenty Simmental dairy cows were considered, 60 grazed on high altitude pasture, and 60 kept indoor and fed a hay-based diet. Cheese production was repeated in two periods (early July and late August) and ripened two and six months. Pasture-derived milk and cheese presented higher fat and lower protein content than hay-derived ones. Rearing systems also affected cheese colour. Textural parameters, hardness, gumminess and chewiness were found to be higher in pasture-derived cheese. In addition, it showed lower level of total saturated fatty acids, and higher level of mono and polyunsaturated fatty acids than hay-derived cheeses. Consumers perceived the difference of cheeses in terms of colour and holes, but they express a similar overall liking. More limited effects of period and ripening time were observed.

Yogurts are important dairy products that have known a rapid market growth over the past few decades. Industrial yogurt manufacture involves different processing steps. Among them, protein fortification of the milk base is elemental. It greatly enhances yogurt nutritional and functional properties and prevents syneresis, an undesirable yogurt textural defect. Protein enrichment can be achieved by either concentration process (evaporation under vacuum and membrane processing: reverse osmosis and/or ultrafiltration) or by addition of dairy ingredients. Traditionally, skim milk powder (SMP) is used to enrich the milk base before fermentation. However, increased quality and availability of other dairy ingredients such as milk protein isolates (MPI), milk protein concentrates (MPC) whey protein isolates (WPI) and concentrates (WPC), micellar casein (MC) and caseinates have promoted their use as alternatives to SMP. Substituting different dry ingredients for skim milk powder in yogurt making affects the yogurt mix protein composition and subsequent textural and sensorial properties. This review focuses on various type of milk protein used for fortification purposes and their influence on these properties.

An experiment was conducted to study the seasonal changes in the fatty acid profile of milk from yaks (Bos grunniens) when kept at altitudes of 3000 m above sea level (a.s.l.) and higher. Data and samples were collected in summer (July), autumn (September), winter (November) and spring (March) from ten lactating yaks (four in spring). The yaks grazed pastures adjacent to the farm building throughout the year. In spring only they received 0·6 kg crop by-products per day (dry matter basis). Fresh alpine grasses, available in summer and autumn, showed high concentrations of α-linolenic acid (46–51 g/100 g lipids) compared with the dry, yellow vegetation of winter and spring (16 g/100 g lipids). In autumn and summer, the milk fat had higher concentrations of polyunsaturated fatty acids than in winter. These polyunsaturated fatty acids were comprised of vaccenic acid, rumenic acid and α-linolenic acid, which are all considered beneficial to human health. The rare fatty acid, γ-linolenic acid, was also detected in yak milk, especially in the milk obtained in spring. The results suggest that yak milk, which is the most important basic food of the Tibetan herders, has the most favourable fatty acid profile when yaks grazed green pasture, which also corresponds to the period of highest milk production.

The total calcium content of cheese, along with changes in the equilibrium between soluble and casein (CN)-bound calcium during ripening can have a major impact on its rheological, functional and textural properties; however, little is known about the effect of other alkaline earth metals. NaCl was partially substituted with MgCl2 or SrCl2 (8·7 and 11·4 g/kg curd, respectively) at the salting stage of cheesemaking to study their effects on cheese. Three cheeses were produced: Mg supplemented (+Mg), Sr supplemented (+Sr) and a control Cheddar cheese. Ca, Mg and Sr contents of cheese and expressible serum obtained therefrom were determined by atomic absorption spectroscopy. Addition of Mg2+ or Sr2+ had no effect on % moisture, protein, fat and extent of proteolysis. A proportion of the added Mg2+ and Sr2+ became CN-bound. The level of CN-bound Mg was higher in the +Mg cheese than the control throughout ripening. The level of CN-bound Ca and Mg decreased during ripening in all cheeses, as did % CN-bound Sr in the +Sr cheese. The presence of Sr2+ increased % CN-bound Ca and Mg at a number of ripening times. Adding Mg2+ had no effect on % CN-bound Ca. The +Sr cheese exhibited a higher G′ at 70 °C and a lower LTmax than the control and +Mg cheeses throughout ripening. The +Sr cheese had significantly lower meltability compared with the control and +Mg cheeses after 2 months of ripening. Hardness values of the +Sr cheese were higher at week 2 than the +Mg and control cheeses. Addition of Mg2+ did not influence the physical properties of cheese. Supplementing cheese with Sr appeared to have effects analogous to those previously reported for increasing Ca content. Sr2+ may form and/or modify nanocluster crosslinks causing an increase in the strength of the para-casein matrix.

The purpose of this work was to analyse the influence of rennet from different Cynara cardunculus plants, selected for its clotting and proteolytic activity on caseins, on the characteristics of manufactured ‘Torta del Casar’ cheeses. After classifying the cardoon according to proteolytic activity into five groups of greater or lesser activity, 16 batches of cheeses were made with rennet derived from different wild cardoon plants. We observed a major development of the proteolysis during ripening leading to the generation of non-protein nitrogen compounds. Especially noteworthy was the relationship of amino acid nitrogen (AN) generation with rennet clotting activity after 24 h of maceration, and the fact that the production of biogenic amines was not related to the proteolytic activity of the rennet. The activities of the rennet observed ‘in vitro’ were also developed ‘in vivo’ in the cheeses, with the different rennets used affecting the final sensory characteristics of cheeses. The rennet with high clotting activity after 24 h of maceration was positively correlated with the creaminess, viscosity, and acceptability of the cheese. However, the high proteolytic activity rennet negatively influenced the acidity, bitterness, and creaminess parameters. Therefore the most appropriate cardoons for making this cheese are those with higher clotting activities and moderate proteolytic activities especially on β-casein. The use of controlled and characterised cardoons in the manufacturing process of Torta del Casar is fundamental to obtaining the homogeneous product demanded by the Torta del Casar Registry of the Protected Designation of Origin.

This study investigated factors affecting milk production and lactation curves from complete lactations of Lacaune dairy sheep. Animals were part of a single flock under intensive management and were milked twice daily starting at lambing. The results of the analyses of 7788 complete lactations showed an average total milk yield of 434±183 l from lactations 234±63 d long, with an average lambing interval of 302±65 d. A Pollott additive mathematical model was used to estimate complete lactation curves. Clustering analysis identified four lactation types among Lacaune dairy sheep differing mainly in productivity i.e. milk yield per lactation (MY) and length of lactation (DIM). The so-called SL type involved short, less productive lactations (n=2137; 27·4%; MY=222±75·5 l and DIM=182±52·9 d). The SN type involved short lactations of normal productivity (n=2039; 26·2%; MY=396±73·7 l and DIM=205±33·1 d). The LP type involved long and productive lactations (n=2169; 27·9%; MY=487±70·5 l and DIM=265±40·7 d), while the LVP type included long and extremely productive lactations (n=1443; 18·5%; MY=694±114·0 l and DIM=295±54·7 d). Sheep showing the best lactation curves were usually younger than other sheep, and they had higher yield during the previous lactation, a shorter previous dry period (55±50·4 for LP and 61±55·0 d for LVP types) and longer lambing intervals. In addition, they tended to be born in September and to lamb in March, October and December. Sheep were remarkably stable in their lactation curve behaviour: the curve type observed for the first lactation was highly likely to persist in subsequent lactations (P<0·0001). These results suggest that farmers can use the shape of the first lactation curve to guide their selection of ewes for breeding and retention on the farm, thereby improving flock productivity.

The aim of this study was to evaluate the effect of feeding goats with distilled and non-distilled thyme leaves (Thymus zygis subsp. gracilis) on the physicochemical composition and technological properties of pasteurised goat milk, and on the physicochemical composition, phenolic content, oxidative stability, microbiology, sensory and texture profile of Murcia al Vino goat cheese. One group of goats was fed the basal diet (control), the second and third groups were fed with different levels of distilled (10 and 20%) or non-distilled (3·75 and 7·5%) thyme leaves. Goat milk physicochemical composition was significantly affected by the substitution of 7·5% of basal goat diet with non-distilled thyme leaves (increase in fat, protein, dry matter and PUFA content), while goat milk clotting time was increased significantly by the introduction of 20% distilled thyme leaves, which reduces its technological suitability. Microbiology, sensory and texture profiles were not affected by the introduction of distilled thyme leaves. The introduction of distilled and non-distilled thyme leaves as an alternative feed to diet can lead to an inhibition of lipids oxidation. The introduction of distilled and non-distilled thyme leaves into goat's diet can be successfully adopted as a strategy to reduce feeding costs and to take advantage of the waste from the production of essential oils, minimising waste removing costs and the environmental impact.

Genotypic and technological properties, antibiotic susceptibility and antimicrobial activity of 35 Leuconostoc strains, isolated from different Italian raw milk cheeses, were investigated. RAPD-PCR was used to study genetic variability and to distinguish closely related strains. The results showed a high degree of heterogeneity among isolates. All the strains had weak acidifying activity and showed low proteolytic and lipolytic activities. Reduction activity, was generally low. All the Leuconostoc were susceptible to ampicillin, mupirocin, erythromycin, quinupristin/dalfopristin and tetracycline. Many strains were classified as resistant to oxacillin, ciprofloxacin and nitrofurantonin, while all isolates were found resistant to vancomycin. PCR-based detection did not identify any of the common genetic determinants for vancomycin (vanA, vanB, vanC1, vanC2, vanC3, vanD, vanE, vanG) or erythromycin (ermB and ermC). Tetracycline resistance genes were detected in 25 tetracycline susceptible strains, the most frequent one being tetM. One strain, belonging to Ln. pseudomesenteroides species, was positive for the presence of the int gene of the Tn916/Tn1545 trasposon family. This is the first time the conjugative transposon Tn916 has been detected inside the Leuconostoc species. All strains showed antimicrobial activity against Enterococcus faecalis and Ent. faecium. The presence of genes encoding amino-acid decarboxylases (hdc and tdc) was not detected. Some strains are interesting in view of their use in cheese production as starter and non starter cultures.

The objective of this study was to collect and analyse milking data from a sample of commercial farms with swingover herringbone parlours to evaluate milking efficiency over a range of parlour sizes (12–32 milking units). Data were collected from 19 farms around the Republic of Ireland equipped with electronic milk metres and herd management software that recorded data at individual milking sessions. The herd management software on each farm was programmed to record similar data for each milking plant type. Variables recorded included cow identification, milking date, identification time, cluster-attachment time, cluster/unit number, milk yield, milking duration, and average milk flow rate. Calculations were performed to identify efficiency benchmarks such as cow throughput (cows milked per h), milk harvesting efficiency (kg of milk harvested per h) and operator efficiency (cows milked per operator per h). Additionally, the work routine was investigated and used to explain differences in the benchmark values. Data were analysed using a linear mixed model that included the fixed effects of season-session (e.g. spring-AM), parlour size and their interaction, and the random effect of farm. Additionally, a mathematical model was developed to illustrate the potential efficiency gains that could be achieved by implementing a maximum milking time (i.e. removing the clusters at a pre-set time regardless of whether the cow had finished milking or not). Cow throughput and milk harvesting efficiency increased with increasing parlour size (12 to 32 units), with throughput ranging from 42 to 129 cows/h and milk harvesting efficiency from 497 to 1430 kg/h (1–2 operators). Greater throughput in larger parlours was associated with a decrease in operator idle time. Operator efficiency was variable across farms and probably dependent on milking routines in use. Both of these require consideration when sizing parlours so high levels of operator efficiency as well as cow throughput can be achieved simultaneously. The mathematical model indicated that application of a maximum milking time within the milking process could improve cow throughput (66% increase in an 18-unit parlour when truncating the milking time of 20% of cows). This could allow current herd milking durations to be maintained as herd size increases.

The aim of this study was to evaluate the effect of storage time (4 °C) on milk samples and the use of azidiol as preservative on the results of microbial inhibitor tests used to detect antimicrobials in milk. For this purpose, 16 milk bulk samples divided into two aliquots, preservative-free and with azidiol, spiked with 12 concentrations of amoxicillin, ampicillin, penicillin G and oxytetracycline, were used. The milk samples were analysed using the BRT MRL, Delvotest MCS Accelerator and Eclipse 100 at 0, 24, 48 and 72 h stored at 4 °C. The logistic regression model was applied to study the effect of storage time (ST), preservative (P) and their interaction (ST×P). At a concentration equivalent to the Detection Limit (DL), the positive results of microbial inhibitor tests do not remain stable during storage time. These results are more reproducible if samples are stored with a preservative. At Maximum Residue Limits (MRLs) concentration microbial inhibitor tests can detect penicillin for up to 72 h of storage. For oxytetracycline, the BRT MRL and Delvotest MCS tests presents DL exceeding the MRL, therefore they are not sensitivity to this tetracycline (0% positive results). By contrast Eclipse 100, whose DL is lower than the MRL, gave 100% positive results during the 72 h storage period. It can be concluded that it would be convenient to store milk samples with azidiol at 4 °C and to carry out analyses within the first 48 h since milk sampling.

The aim of the present study was the examination of the boot swab sampling technique for the collection of environmental material in order to identify Mycobacterium avium ssp. paratuberculosis (MAP)-infected herds. Eight dairy herds were included into the study. Four of them had a well-known history of MAP-infection from a herd surveillance programme conducted since 2006. Cows in these herds were repeatedly tested positive in Pourquier® MAP-ELISA (Pourquier, Montepellier, France); in some MAP could be isolated in individual faecal culture despite that symptoms of paratuberculosis were never reported. In four presumably negative herds nearly all cows were repeatedly tested serologically negative for MAP. The pathogen was never isolated from faecal samples of cows by culture. The study was initiated with the aim of standardising environmental samples as a herd diagnostics, in which overall 130 pairs of boot swab samples from the cows’ surroundings were taken In 58 of 64 swab samples (90·6%) from confirmed MAP-infected herds the organism could be isolated by mycobacterial culture of the boot swab. Contrarily, in 66 samples from presumably MAP-negative herds only one swab was positive (1·5%). The utilisation of boot swabs as a standardised technique for environmental sampling offers an effective and inexpensive tool for identifying herds infected with MAP. This is the first report of using boot swabs for the collection of environmental samples for MAP- detection in cattle herds. This easy to perform technique enables the economical detection of MAP herd status.

The dairy industry uses lipase extensively for hydrolysis of milk fat. Lipase is used in the modification of the fatty acid chain length, to enhance the flavours of various chesses. Therefore finding the unlimited source of lipase is a concern of dairy industry. Due to the importance of lipase, this study was an attempt to express the lipase from Burkholderia cepacia in Lactococcus lactis. To achieve this, a gene associated with lipase transport was amplified and subcloned in inducible pNZ8148 vector, and subsequently transformed into Lc. lactis NZ9000. The enzyme assay as well as SDS-PAGE and western blotting were carried out to analysis the recombinant lipase expression. Nucleotide sequencing of the DNA insert from the clone revealed that the lipase activity corresponded to an open reading frame consisting of 1092 bp coding for a 37·5-kDa size protein. Blue colour colonies on nile blue sulphate agar and sharp band on 37·5-kD size on SDS-PAGE and western blotting results confirm the successful expression of lipase by Lc. lactis. The protein assay also showed high expression, approximately 152·2 μg/ml.h, of lipase by recombinant Lc. lactis. The results indicate that Lc. lactis has high potential to overproduce the recombinant lipase which can be used commercially for industrially purposes.

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.