The aim of this research was to investigate variability in each of the five exons of the caprine growth hormone (gGH) gene, in order to establish the possible relationships with milk traits in Sarda breed goat. The general linear model procedure was used to analyse the effects of the single strand conformation (SSCP) profiles on milk traits of 100 lactating goats. Analysis of conformational polymorphism at exons 1–5 revealed a total of 25 differing banding patterns. Sequencing revealed 21 nucleotide changes (compared with GenBank D00476): 14 were polymorphic and 7 monomorphic; 19 in exonic regions, 5 of which were nonsynonymous. A SNP upstream of the transcription initiation codon (c.-3A>G) and an indel (c.*29_30insC) in the 3′UTR, were detected. Alignment of 4 cloned sequences including the entire gGH gene led to the identification of 22 nucleotide variations within the intron regions, including two indels. Association analysis revealed that each exon, except exon-1, affected milk yield, exons 1 and 3 influenced milk fat percentage, and all exons, except exon-2, had an effect on protein percentage, supporting previous results in livestock. The variability detected at the caprine GH gene might provide useful information for the phylogeny of ruminants and, more importantly, have implications on the biological function of the growth hormone and on those traits resulting from its physiological action, including milk production and composition. The caprine GH gene may become a useful molecular marker for a more effective genetic selection for milk production traits in goats.

The effect of CSN1S1 genotype and lactation stage on milk yield and composition were investigated in 80 extensively reared goats. Milk yield was recorded in early, mid and late lactation and individual milk samples were collected to determine: fat, protein, lactose and casein content, pH, freezing point, somatic cell count (SCC) and total microbic mesophilic count (TMC). Relative casein composition and amino acid profile were quantified by HPLC. Fatty acid profile was measured by gas-chromatography. Genotype did not affect milk yield, while this trait was significantly affected by lactation stage (P < 0·01). CSN1S1 BB goats produced significantly higher protein and casein percentages (P < 0·05). αs1-casein (CN) was significantly higher in BB and AB goats than AF and BF, showing intermediate values in AA goats (P < 0·01). The protein percentage and the αs1 and αs2-CN fractions were not affected by lactation stage, while the casein content and the β and κ-CN significantly increased throughout lactation (P < 0·01). C4 : 0 and C6 : 0 were not affected by genotype, while C8 : 0 and C10 : 0 were higher in the AA goats than BB; most of the long chain FA were higher in BB than AA goats. MUFA and PUFA increased in late lactation. In addition, BB goats showed higher essential amino acids, resulting in an optimal composition from the nutritional point of view, when compared with AA goats. The increase of MUFA, PUFA, essential and cis-FA in late lactation indicate that the lipid composition of goat's milk, with the progress of lactation, tends to improve its nutritional value.

The aim of this study was to assess the effect of chemical composition, coagulation properties, pH, and titratable acidity (TA, SH°/50 ml) of vat milk on Grana Padano cheese yield (CY) under field conditions. Twelve cheese-making sessions were carried out from February to December 2009 in a dairy cooperative of Grana Padano Consortium (Italy), for a total of 96 vats of milk processed. For each vat, samples of raw milk were collected and analysed for quality traits (fat, protein, and casein contents), pH, TA, and milk coagulation properties (MCP), measured as rennet coagulation time (RCT, min), curd-firming time (k20, min), and curd firmness (a30, mm). Cheese yield was expressed as kilograms of cheese per 100 kg milk transformed, and was measured after 2 d of drainage. Fat, protein, and casein contents were positively and strongly correlated with CY (coefficients of correlation, r = 0·72, 0·88, and 0·84, respectively; P < 0·001). Coagulation properties were moderately and significantly (P < 0·001) related to CY: milk that coagulated earlier and had stronger a30 was associated to greater CY. Cheese yield was analysed with a model that accounted for fixed effects of cheese-making day, fat and protein content, TA, and a30. Significance was found for all the effects (P < 0·05). Milk characterised by high values of a30 resulted in higher CY than milk with low values of a30, indicating that MCP could be used as indicators of cheese-making efficiency. Future research should investigate the relationships between MCP and quality of cheese, and explore the feasibility of including MCP in multiple component milk pricing system for Grana Padano cheese production.

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.

Gelation is a significant operation in dairy processing. Protein gelation can be affected by several factors such as temperature, pH, or enzyme addition. Recently, the use of ultrasonication has been shown to have a significant impact on the formation of whey protein gels. In this work, the effect of ultrasonication on the gelation of casein systems was investigated. Gels were formed by the addition of 7·6 mm Tetra Sodium Pyro Phosphate (TSPP) to 5 wt% micellar casein (MC) solutions. Sonication at 20 KHz and 31 W for up to 30 min changed the surface hydrophobicity of the proteins, whereas surface charge was unaltered. Sonication before the addition of TSPP formed a firm gel with a fine protein network and low syneresis. Conversely, sonication after TSPP addition led to an inconsistent weak-gel-like structure with high syneresis. Gel strength in both cases increased significantly after short sonication times, while the viscoelastic properties were less affected. Overall, the results showed that ultrasonication can have a significant effect on the final gel properties of casein systems.

The aim of this study was to evaluate the ability of Bifidobacterium strains to prevent the effects associated with Clostridium difficile infection in a hamster model of enterocolitis. After clindamycin treatment (30 mg/kg), animals were infected intragastrically with C. difficile (5×108 CFU per animal). Seven days prior to antibiotic administration, probiotic treatment was started by administering bacterial suspensions of bifidobacteria in drinking water. Strains CIDCA 531, CIDCA 5310, CIDCA 5316, CIDCA 5320, CIDCA 5323 and CIDCA 5325 were used. Treatment was continued during all the experimental period. Development of diarrhoea, enterocolitis and mortality were evaluated. All the infected animals belonging to the placebo group developed enterocolitis (5/5) and only two dead (2/5) whereas in the group administered with Bifidobacterium bifidum strain CIDCA 5310 the ratio of animals with enterocolitis or dead decreased significantly (1/5 and 0/5 respectively). Biological activity of caecum contents was evaluated in vitro on Vero cells. Animals treated with strain CIDCA 5310 presented lower biological activity than those belonging to the placebo group. The present study shows the potential of selected strains of bifidobacteria to antagonise, in vivo, the virulence of C. difficile.

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.

Product authenticity is an important topic in dairy sector. Dairy products sold for public consumption must be accurately labelled in accordance with the contained milk species. Linear discriminant analysis (LDA), a common chemometric procedure, has been applied to fatty acid% composition to classify pure milk samples (cow, ewe, buffalo, donkey, goat). All original grouped cases were correctly classified, while 90% of cross-validated grouped cases were correctly classified. Another objective of this research was the characterisation of cow-ewe milk mixtures in order to reveal a common fraud in dairy field, that is the addition of cow to ewe milk. Stereospecific analysis of triacylglycerols (TAG), a method based on chemical–enzymatic procedures coupled with chromatographic techniques, has been carried out to detect fraudulent milk additions, in particular 1, 3, 5% cow milk added to ewe milk. When only TAG composition data were used for the elaboration, 75% of original grouped cases were correctly classified, while totally correct classified samples were obtained when both total and intrapositional TAG data were used. Also the results of cross validation were better when TAG stereospecific analysis data were considered as LDA variables. In particular, 100% of cross-validated grouped cases were obtained when 5% cow milk mixtures were considered.

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.

Propionibacteria derived from dairy products are relevant starter cultures for the production of Swiss and Emmental-type cheeses, and the monitoring of which is mandatory for proper quality control. This study aimed to evaluate an alternative procedure to enumerate propionibacteria, in order to develop a reliable and practical methodology to be employed by dairy industries. 2,3,5-triphenyltetrazolium chloride (TTC) inhibitory activity was tested against five reference strains (CIRM 09, 38, 39, 40 and 116); TTC at 0·0025% (w/v) was not inhibitory, with the exception of one strain (CIRM 116). Subsequently, the four TTC-resistant strains, three commercial starter cultures (PS-1, PB-I, and CHOO) and twelve Emmental-type cheese samples were subjected to propionibacteria enumeration using Lithium Glycerol (LG) agar, and Petrifilm™ Aerobic Count (AC) plates added to LG broth (anaerobic incubation at 30 °C for 7 d). Petrifilm™ AC added to LG broth presented high counts than LG agar (P<0·05) for only two reference strains (CIRM 39, and 40) and for all commercial starter cultures. Cheese sample counts obtained by both procedures did not show significant differences (P<0·05). Significant correlation indexes were observed between the counts recorded by both methods (P<0·05). These results demonstrate the reliability of Petrifilm™ AC plates added to LG broth in enumerating select Propionibacterium spp., despite some limitations observed for specific commercial starter cultures.

The effect of partial substitution of NaCl with KCl on texture profile, soluble Ca, K, Na, and P, and microstructure of low-moisture mozzarella cheese (LMMC) was investigated. LMMC batches were prepared using four combinations of NaCl and KCl salt viz., NaCl only, NaCl:KCl, 3:1, 1:1 and 1:3 (w/w); all used at of 46 g/kg curd and plasticised in 4% brine containing the above salt mixtures. Texture profile, microstructure, and percentages of soluble Ca, K, Na, and P were determined. There were no significant differences in hardness, cohesiveness, adhesiveness, and gumminess among the experimental LMMC batches. Environmental scanning electron microscopy images showed compact and homogeneous structure of LMMC at day 27 of storage; however, no significant difference was observed among the experimental LMMC batches. Hardness increased significantly in all experimental LMMC during storage. LMMC salted with NaCl/KCl mixtures had almost similar sensory properties compared with the control. There was no significant difference in creaminess, bitterness, saltiness, sour-acid, and vinegary taste among the experimental LMMC at the same storage period.

The present study examines the resistance of the α-lactalbumin to α-chymotrypsin (EC 3.4.21.1) digestion under various experimental conditions. Whey protein isolate (WPI) was hydrolysed using randomised hydrolysis conditions (5 and 10% of WPI; pH 7·0, 7·8 and 8·5; temperature 25, 37 and 50 °C; enzyme-to-substrate ratio, E/S, of 0·1%, 0·5 and 1%). Reversed-phase high performance liquid chromatography (RP-HPLC) was used to analyse residual proteins. Heat, pH adjustment and two inhibitors (Bowman–Birk inhibitor and trypsin inhibitor from chicken egg white) were used to stop the enzyme reaction. While operating outside of the enzyme optimum it was observed that at pH 8·5 selective hydrolysis of β-lactoglobulin was improved because of a dimer-to-monomer transition while α-la remained relatively resistant. The best conditions for the recovery of native and pure α-la were at 25 °C, pH 8·5, 1% E/S ratio, 5% WPI (w/v) while the enzyme was inhibited using Bowman–Birk inhibitor with around 81% of original α-la in WPI was recovered with no more β-lg. Operating conditions for hydrolysis away from the chymotrypsin optimum conditions offers a great potential for selective WPI hydrolysis, and removal, of β-lg with production of whey protein concentrates containing low or no β-lg and pure native α-la. This method also offers the possibility for production of β-lg-depleted milk products for sensitive populations.

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.

Curcumin is a bioactive polyphenolic compound extracted from turmeric with known anti-inflammatory properties, and its hydrophobic nature restricts its solubility and its bioaccessibility. Solubility may be improved upon binding of curcumin to native or treatment-modified casein micelles. The present work demonstrated that high hydrostatic pressure treatment of skim milk increases the binding of curcumin to caseins. The association of curcumin to casein micelles was assessed using fluorescence spectroscopy, either directly or by tryptophan quenching. The amount of curcumin associated with the milk proteins increased in pressure-treated milk, and a further improvement in the amount of bound curcumin was observed upon pressure treatment of a milk/curcumin mixture. However, in this case, some of the curcumin dissociated during storage, contrarily to what was observed for untreated milk. From a molecular standpoint, the data presented here indicate that structural modifications induced by high-pressure treatment and known to affect the structure of milk proteins result in a rearrangement of the amino acid residues in close proximity to the protein-associated curcumin.

The effects of adding flaxseed or fish oil to the diet of dairy cows on the chemical and physical profile of Mozzarella cheese production were studied. The experiment involved 24 Friesian cows, divided into 3 groups accordingly fat supplementation: basal diet (CT), diet supplemented with flaxseed (FS) or fish oil (FO). Mozzarella cheeses were manufactured from bulk milk of each group. Bulk milk was analysed for chemical composition and renneting parameters. Mozzarella cheeses were analysed for chemical composition, fatty acid profile, and textural properties. Results suggest that Mozzarella cheese from cows receiving flaxseed supplementation showed a decrease in saturated fatty acids (SFA), an increase in monounsaturated fatty acids (MUFA), and in polyunsaturated fatty acids (PUFA) compared with control Mozzarella cheese. The increased dietary intake of C18:3 in flaxseed supplemented cows resulted in increased levels of trans-11 C18:1, and of CLA cis-9 trans-11 C18:2, and in low Atherogenic and Trombogenic Indexes. FO Mozzarella cheese showed compositional and textural properties quite similar to CT Mozzarella cheese; however, increased levels of n-3 polyunsaturated fatty acids in FO Mozzarella were found.

Conventional medical therapies for ulcerative colitis (UC) are still limited due to the adverse side effects like dose-dependent diarrhoea and insufficient potency to keep in remission for long-term periods. So, new alternatives that provide more effective and safe therapies for ulcerative colitis are constantly being sought. In the present study, probiotic LaBb Dahi was selected for investigation of its therapeutic effect on DSS-induced colitis model in mice. LaBb Dahi was prepared by co-culturing Dahi culture of Lactococci along with selected strain of Lactobacillus acidophilus LaVK2 and Bifidobacterium bifidum BbVK3 in buffalo milk. Four groups of mice (12 each) were fed for 17 d with buffalo milk (normal control), buffalo milk plus DSS (Colitis control), Dahi plus DSS, and LaBb Dahi plus DSS, respectively, with basal diet. The disease activity scores, weight loss, organ weight, colon length, myeloperoxidase (MPO) and β-glucoronidase activity was assessed, and the histopathological picture of the colon of mice was studied. All colitis control mice evidenced significant increase in MPO, β-glucoronidase activity and showed high disease activity scores along with histological damage to colonic tissue. Feeding with LaBb Dahi offered significant reduction in MPO activity, β-glucoronidase activity and improved disease activity scores. We found significant decline in length of colon, organ weight and body weight in colitis induced controls which were improved significantly by feeding LaBb Dahi. The present study suggests that LaBb Dahi can be used as a potential nutraceutical intervention to combat UC related changes and may offer effective adjunctive treatment for management of UC.

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 effect of high-pressure treatment (400, 500 and 650 MPa) on the structure and activity of bovine lactoferrin in different iron-saturation forms has been studied by several techniques. The structural changes produced in lactoferrin by high-pressure were analysed by differential scanning calorimetry and fluorescence spectroscopy, and the immunoreactivity by ELISA. The effect of high-pressure was also studied on some biological properties of lactoferrin, such as iron binding capacity, retention of the bound iron, and antibacterial activity against Escherichia coli O157:H7. Results obtained indicate that treatment at 400 MPa does not substantially modify the conformation of lactoferrin, meanwhile treatments at 500 and 650 MPa greatly affect some of its properties. With respect to the antibacterial activity, the apo and native forms of lactoferrin maintain that activity against Esch. coli only after 400 MPa treatment.

Several mastitis-causing pathogens are able to colonize the bovine teat canal. The objective of this study was to investigate the association between the treatment of sawdust bedding with a commercial alkaline conditioner and the bacterial counts on teat skin and in the teat canal. The study used a crossover design. Ten lactating Holstein cows that were free of udder infections and mastitis were included in the study. The animals were bedded on either untreated sawdust or sawdust that had been treated with a hydrated lime-based conditioner. Once a day, fresh bedding material was added. After 3 weeks, the bedding material was removed from the cubicles, fresh bedding material was provided, and the cows were rotated between the two bedding material groups. Teat skin and teat canals were sampled using the wet and dry swab technique after weeks 1, 2, 3, 4, 5 and 6. Staphylococcus aureus, Streptococcus uberis, Escherichia coli and other coliform bacteria were detected in the resulting agar plate cultures. The treatment of the bedding material was associated with the teat skin bacterial counts of Str. uberis, Esch. coli and other coliform bacteria. An association was also found between the bedding material and the teat canal bacterial counts of coliform bacteria other than Esch. coli. For Staph. aureus, no associations with the bedding material were found. In general, the addition of a hydrated lime-based conditioner to sawdust reduces the population sizes of environmental pathogens on teat skin and in teat canals.