Emulsions were prepared with 20% soy oil and different concentrations of lactoferrin, and tested at pH values from 3 to 7·5. The stability of the emulsions decreased as the pH got closer to the isoelectric point of the protein. A concentration of 1% lactoferrin was determined to be sufficient to provide full coverage of the emulsion droplets. Lactoferrin-stabilized emulsions were then prepared in water at pH 6·6 and their behaviour when added to reconstituted milk was studied. It was observed that lactoferrin emulsions were stable when reconstituted in milk, but they showed aggregation when diluted in milk serum alone. The destabilization was caused by shielding of the charges on the surface of the oil droplets. Stabilization in milk occurred due to interactions at the interface with other soluble proteins. In fact, when β-lactoglobulin or sodium caseinate were added to the serum, stability of the emulsion droplets was restored, indicating that these proteins were able to adsorb at the interface and aid in the stabilization. Since ζ-potential measurements did not show significant overall charge on the emulsion droplets, this suggests that the stabilization forces are not only electrostatic in nature, but that there are other mechanisms at play.

This study evaluated the effects of probiotic Dahi administration in ageing mice on macrophage and lymphocyte functions. Probiotic Dahi were prepared by co-culturing in buffalo milk (3% fat) Dahi bacteria (Lactococcus lactis ssp. cremoris NCDC-86 and Lc. lactis ssp. lactis biovar diacetylactis NCDC-60) along with Lactobacillus acidophilus LaVK2 (La-Dahi) or combined Lb. acidophilus and Bifidobacterium bifidum BbVK3 (LaBb-Dahi). Four groups of 12 mo old mice were fed for four months, with the supplements (5 g/day) of buffalo milk (3% fat), Dahi, La-Dahi and LaBb-Dahi, respectively, in addition to basal diet, and a fifth group that received no supplements served as control. The immune functions of young mice (4 mo old) were also compared with those of ageing adult mice (16 mo old). The production of nitric oxide and cytokines IL-6 and TNF-α declined and that of immunosuppressive prostaglandin E2 (PGE2) increased by stimulated peritoneal and splenic macrophages in ageing mice, compared with their young counterparts. The proliferation of stimulated splenocytes diminished and the production of IL-2 decreased and that of IL-6 and TNF-α enhanced in ageing compared with young mice. Feeding ageing mice with La-Dahi or LaBb-Dahi improved peritoneal macrophage functions stimulating nitric oxide and IL-6 and diminishing PGE2 production. Feeding La-Dahi or LaBb-Dahi also improved lymphocyte functions stimulating their proliferation and production of IL-2 in ageing mice. To conclude, the probiotic La-Dahi and LaBb-Dahi are effective in reversing age related decline in immune functions in mice.

Samples of the mammary gland were obtained by the freeze-clamping technique from heifers, lactating and dry cows. Levels of acetyl CoA, citrate, isocitrate and α-oxoglutarate were determined in extracts of the samples and calculated per g fresh gland and per mg DNA. All mammary metabolite levels were significantly higher in lactating cows than in heifers or dry cows. The accumulation in the lactating gland increased in the sequence acetyl CoA–α-oxoglutarate–isocitrate–citrate. NADP/NADPH ratios in cytoplasm, as calculated from the equilibrium of isocitrate dehydrogenase, were lowest in lactating glands. The activities of some enzymes were also determined in the cytoplasm and calculated per g fresh gland, per mg protein and per mg DNA. Mammary citrate synthase activity did not vary significantly between different groups of cattle, whereas mammary isocitrate dehydrogenase activity was higher in lactating cows than in heifers or dry cows. No activity of ATP citrate lyase was detected.

The aim of this study was to compare seven non-linear mathematical models (Brody, Wood, Dhanoa, Sikka, Nelder, Rook and Dijkstra) to examine their efficiency in describing the lactation curves for milk fat to protein ratio (FPR) in Iranian buffaloes. Data were 43 818 test-day records for FPR from the first three lactations of Iranian buffaloes which were collected on 523 dairy herds in the period from 1996 to 2012 by the Animal Breeding Center of Iran. Each model was fitted to monthly FPR records of buffaloes using the non-linear mixed model procedure (PROC NLMIXED) in SAS and the parameters were estimated. The models were tested for goodness of fit using Akaike's information criterion (AIC), Bayesian information criterion (BIC) and log maximum likelihood (−2 Log L). The Nelder and Sikka mixed models provided the best fit of lactation curve for FPR in the first and second lactations of Iranian buffaloes, respectively. However, Wood, Dhanoa and Sikka mixed models provided the best fit of lactation curve for FPR in the third parity buffaloes. Evaluation of first, second and third lactation features showed that all models, except for Dijkstra model in the third lactation, under-predicted test time at which daily FPR was minimum. On the other hand, minimum FPR was over-predicted by all equations. Evaluation of the different models used in this study indicated that non-linear mixed models were sufficient for fitting test-day FPR records of Iranian buffaloes.

Bovine, ovine and caprine pepsin and lamb, piglet and kid chymosins (rennins) were isolated by basically the same procedure of selective pH precipitation, DEAE-cellulose chromatography in phosphate buffer, pH 6·5 and Sephadex G-100 chromatography. The homogeneity of the preparations was assessed by (a) re-cycling on Sephadex G-100, (b) chromatography on DEAE-cellulose using piperazine buffer, pH 5·3 and (c) ability to inactivate ribonuclease.

Three rations were compared in a 15-week winter feeding experiment using 12 Ayrshire cows. All the cows received silage 3 times a day and concentrates; ration A contained no sugar-beet pulp whilst ration B contained 6 lb of pulp and ration C 12 lb.

The total weights of dry matter consumed were 29·7, 32·3 and 33·4 lb/day, respectively, on treatments A, B and C. For every 1 lb of extra sugar-beet pulp dry matter eaten the decline in the dry weight of the other feeds consumed was 0·50 lb on treatment B and 0·55 lb on treatment C. The main decrease was in the silage part of the ration. The weights of starch equivalent (S.E.) consumed daily were 18·0, 19·8 and 20·7 lb on rations A, B and C, respectively, with mean daily milk yields of 38·2, 39·3 and 39·7 lb, giving a response of 0·55 lb milk/lb extra S.E. between rations A and C. The corresponding fat percentages of the milks did not differ significantly from each other but the solids-not-fat (SNF) percentages increased significantly from 8·76% on ration A to 8·88% on ration C. The average response per lb extra S.E. was 0·044% SNF and the increase was in the protein fraction of the milk. The cows were significantly heavier on rations B and C than on ration A. The inclusion of beet pulp in rations B and C had no adverse effect on the taste of the milk.

Bacteriological examination of 15 pairs of Cheddar cheeses, of which one of each pair was made aseptically, showed that there was a relationship between the development of rancid flavour and the numbers of Clostridium tyrobutyricum present. Other species of clostridia, Cl. beijerinkii, Cl. sporogenes and Cl. perfringens, were also present but occurred only as spores. Conditions for the growth of Cl. tyrobutyricum appeared to be more favourable in the aseptically made cheese; it is suggested that the adventitious bacterial flora of the non-aseptically made cheese, which gains access during cheesemaking, has some inhibitory effect on the growth of this organism.

We investigated whether different rearing conditions affected the physical development, ease of integration into the dairy herd and performance of primiparous dairy cows and the results are reported in this Research Communication. The three rearing conditions investigated were whole-day cow-calf contact for 9 weeks (WDC), half-day contact for nine weeks (HDC) and no mother-contact (NC) with nipple-bucket-feeding (max. 2 × 3 l/d) and group-housing from the 8th day onward. After permanent separation from the dams (WDC and HDC), all calves had been nipple-bucket-fed and gradually weaned from week 10 to 13 of life and kept together as calves and heifers. Measures of physical development were trunk girth, height at withers and body weight. Lying behavior during the first 48 h after introduction to the dairy herd after first calving was used as an indicator of ease of integration. Performance measures were age at calving, lactation duration, milk yield and culling rates during the first lactation. No differences between WDC and HDC could be detected. Moreover, no treatment-effects on physical development or performance could be found. All treatments showed reduced lying for the first 24 h after introduction to the dairy herd. Afterward NC-heifers lay less than WDC, with HDC-heifers ranging in between. We conclude that under the conditions investigated higher weights two weeks after weaning in WDC- and HDC-calves did not carry through to the first lactation and did not lead to earlier maturity and higher performance, but that integration into the herd may be alleviated when calves have early experience of the herd and associated conditions.

Dulce de leche is a dairy-based confectionery product, widely consumed in Argentina and other Latin American countries as filling or topping for desserts, cakes and biscuits (Hansen, 1978; Bouzas, 1997). It is prepared by heat concentration of whole milk with added sucrose until it reaches 700 g total solids/kg dulce de leche. Sucrose is usually partly replaced by other sugars (not more than 400 g/kg of total sugars added according to Normas Mercosur, 1996) to avoid crystallization. Sodium bicarbonate is added to prevent coagulation of proteins as the pH of the mixture decreases and the isoelectric pH of proteins increases during heating.

Maillard browning is the main reaction of milk proteins during the manufacture of dulce de leche. This results in an attractive flavour and colour, but also leads to less desirable changes, such as loss of nutritional value and formation of toxic compounds (Rizzi, 1994; Friedman, 1996a). The natural reactants are free amino groups of milk proteins and lactose. However, as the addition of other reducing sugars is an accepted practice, the consequences may be quite different depending on the formulation used. In addition, other reactions leading to crosslinkages between protein chains contribute to the reduction in nutritional value (Mauron, 1990; Friedman, 1996b; Finot, 1997). The reduction of nutritional quality of milk proteins is attributed to the lower digestibility and the biological inactivation or destruction of essential amino acids. Lysine is the essential amino acid most affected as its free ε-amino group is very reactive (Hurrell, 1990; Mauron, 1990; Finot, 1997).

Several studies have investigated the physical and rheological characteristics (Pauletti et al. 1984, 1996), sensory properties (Sabioni et al. 1984; Buera et al. 1990; Hough et al. 1990), sugar composition and microbiological stability (Ferramondo et al. 1984) of dulce de leche. However, there is little information about the loss in available lysine during manufacture (Pavlovic et al. 1994).

The aim of this study was to examine the effects of the nature and quantity of various sugars commonly used in the manufacture of dulce de leche on the loss of available lysine.

Investigations on the growth characteristics of Streptococcus lactis ML8 in skim-milk medium have indicated that agitation of the medium may decrease the growth rate of the organism by decreasing the availability of CO2 (CO2 effect) and of soluble nitrogen (soluble nitrogen effect). The former effect was observed only when the CO2 concentration was depleted by heat sterilization; the latter occurred when a certain threshold level of agitation was exceeded. Results suggested that the soluble nitrogen affect was caused by an alteration in the rate of diffusion of substances formed in the immediate vicinity of the bacterial cell by the proteolytic enzyme system.

Milk protein synthesis in the mammary gland involves expression of six major milk protein genes whose nutritional regulation remains poorly defined. In this study, the effect of long term under- and over-feeding on the expression of αs1-casein: CSN1S1, αs2-casein: CSN1S2, β-casein: CSN2, κ-casein: CSN3, α-lactalbumin: LALBA and β-lactoglobulin: BLG gene in sheep mammary tissue (MT) was examined. Twenty-four lactating dairy sheep, at 90–98 d in milk, were divided into three groups and fed the same ration, for 60 d, in quantities which met 70% (underfeeding), 100% (control) and 130% (overfeeding) of their energy and crude protein requirements. The results showed a significant reduction on mRNA of CSN1S1, CSN1S2, CSN2 and BLG gene in the MT of underfed sheep compared with the overfed ones and a significant reduction in CSN3 and LALBA gene expression compared with the respective control animals. Significant positive correlations were observed between the mRNA levels of milk proteins’ genes with the milk protein yield and milk yield respectively. In conclusion, the feeding level and consequently the nutrients availability, affected the milk protein yield and milk volume by altering the CSN1S1, CSN1S2, CSN2, CSN3, LALBA and BLG gene expression involved in their metabolic pathways.

The objective of this study was to determine whether Staphylococcus aureus chronic intramammary infection (IMI) influences expression of proteins related to regulation of proliferation and apoptosis processes and proliferation/apoptosis index during active involution in bovine mammary gland. Twenty-one Holstein non-pregnant cows in late lactation either uninfected or with chronic naturally acquired S. aureus IMI were included in this study. Cows were slaughtered at 7, 14 and 21 d after cessation of milking and samples for immunohistochemical analysis were taken. Protein expression of Bcl-2, Bax, Fas and active caspase-3 in mammary tissue was significantly affected by chronic S. aureus IMI, all showing increased immunoexpression in S. aureus-infected quarters at all involution stages. The percentage of apoptotic cells was increased by IMI in both mammary parenchyma and stroma, and the percentage of parenchymal and stromal cell proliferation was also increased. The proliferation/apoptosis ratio was significantly increased by IMI only in stromal cells. This imbalance to favour proliferation in S. aureus-infected mammary quarters could be one of the underlying causes that induce aberrant involution with permanence of nonsecretory tissue and increase of stromal components.

Loss of milk clotting activity when rennin was treated with urea at several concentrations and at different pH values was compared with the change in UV absorption of the protein. The change in absorption at 287 and 293 nm did not correlate with loss in activity, indicating that the loss of enzymic properties was not related to the extent of unfolding of the molecule but was related to the reaction of susceptible groups to the environmental conditions.

Reactions of modifying agents, for example N-bromoacetamide, 2-hydroxy-5-nitrobenzyl bromide and mercuric ions with tryptophan groups and the relationship of these changes to enzyme activity suggested that tryptophan was not specifically associated with enzyme action but was important in maintaining tertiary structure.

Rennin was not inhibited by the pepsin inhibitors p-bromophenacyl bromide and α-diazo-p-bromoacetophenone which suggested that the active site did not possess a reactive carboxyl group as in pepsin. However, the reversible inhibition with 1-cyclohexyl-3(2morpholinoethyl)-carbodiimide metho p-toluenesulphonate together with the results of oxidation by N-bromoacetamide indicated that a tyrosine group or groups was important in enzyme activity.

Angiotensin-I-Converting Enzyme (ACE) inhibitors peptides were produced from unsupplemented acid goat whey fermented aerobically for 168 h by Kluyveromyces marxianus and Lactobacillus rhamnosus. This yeast-lactobacillus association is GRAS. Two novel lactokinins were identified: NYW and W with IC50 of 20 and 0·86 μm respectively. They both were resistant toward simulated gastrointestinal digestion. In addition, WLAHK was found in the hydrolysate. These three sequences belong to f(99–110) of α-la which seems to be a lactokinin cryptic zone. W was the major molecule released by the fermentation process. Considering that W is the precursor of serotonin, the hydrolysate produced could be of interest for the generation of functional health ingredient involved in regulation of affective disorders and hypertension.

The rate of removal of a dissolved tainting substance from cream by steam-distillation treatment is dependent in part on the relative volatility (or boiling point) of the pure substance, in part on the temperature of steam distillation, and in part on the influence of water on the vapour pressure of the tainting substance in dilute solution (i.e. on the activity coefficient, γ).

The application of Raoult's law and Henry's law to the problem of removal of taints from cream is discussed. For the dilute solutions commonly encountered in cream Henry's law can be expected to apply.

The equilibrium relationships for diacetyl and acetoin used as reference substances in investigations on cream-deodorizing equipment have been determined. The equilibrium coefficient for diacetyl in water at concentrations up to 50 p.p.m. in the liquid, as measured by the continuous equilibrium vaporization method, is 39·3. With rise in concentration above 50 p.p.m. in the liquid the coefficient decreases. At 100 p.p.m. it is 32 and at 1000 p.p.m. it is 20. The coefficient for acetoin in water at all concentrations up to 1·2% in the liquid is constant at 1·29. The activity coefficient for diacetyl in concentrations up to 50 p.p.m. was found to be 26·4 and for acetoin at concentrations up to 1·2%, 5·4.

For diacetyl the solubility partition coefficient as between butterfat and water rises from 0·47 at 100° F. to 0·74 at 180° F. For acetoin the partition coefficient remains constant at 0·18–0·19 over this range of temperatures. Correspondingly diacetyl is removed equally easily from butterfat and from water, whereas acetoin steam-distils much more readily from butterfat than from water.

The objective of the study reported in this Research Communication was to investigate the association of polymorphisms in the insulin-like growth factor receptor 2 (IGF2R) gene with milk traits in 283 Polish Holstein-Friesian (PHF) cows from the IGAB PAS farm in Jastrzębiec. IGF2R regulates the availability of biologically active IGF2 which is considered as a genetic marker for milk or meat production in farm animals. Two novel genetic polymorphisms were identified in the bovine IGF2R gene: a polymorphic TG-repeat in intron 23 (g.72389 (TG)15-67), and a g.72479 G > A SNP RFLP-StyI in exon 24. The following milk traits were investigated: milk yield, protein and fat yield, SCC and lactose content. To determine the influence of the IGF2R STR and SNP genotypes on the milk traits, we used the AI-REML (average information restricted maximum likelihood) method with repeatability, multi-trait animal model based on test-day information using DMU package. Statistical analysis revealed that the G/A genotype (P ≤ 0·01) was associated with milk and protein yield, lactose content and somatic cell count (SCC) in Polish HF cows. TGn (29/22, 28/29, 28/22, 28/28) genotypes were associated with high values for milk, (28/22, 28/23) with protein and fat yield, (25/20) with lactose content, and (29/33, 28/28) with low SCC. We suggest that the IGF2R gene polymorphisms could be useful genetic markers for dairy production traits in cattle.

A method for the determination of the fat content of milk from measurements of the turbidities of diluted milk suspensions in the near infra-red region has been shown to provide greater accuracy than the alkali-addition method in the visible region. Variations in results due to breeds of cows and lactation times have been studied, and excluding early lactation samples, the maximum deviation within a given breed was found to correspond to ±0·2% in the fat concentration.

At present there is no rapid microbiological method for the detection of viable Mycobacterium paratuberculosis in milk. By combining an extensive milk sample pretreatment with solid phase cytometry as the detection technique we were able to demonstrate viable mycobacterial cells in 50 ml of artificially contaminated pasteurized milk in less than one working day.

Ectoine is a compatible solute accumulated in halophilic bacteria in response to high salt concentrations and offers protection from osmotic stress. The occurrence of compatible solutes is widespread among bacteria, yet ectoine has never been detected in foods. The use of an ectoine producing microorganism (Brevibacterium linens) in the surface ripening of red smear cheeses led to the question whether ectoine can be found in cheese. Therefore we examined samples from a variety of cheese manufacturers and different types of red smear cheeses for the presence of ectoine using HPLC and HPLC/MS analysis. Ectoine solely appears in the rind and was detected up to 178 mg/200 g red smear cheese, depending on several factors like ripening status and conditions throughout the cheese production process (e.g. salt concentrations of used brine baths).