Sorting of normal and abnormal milk at time of milking is done visually for conventional milking systems, but more concrete standards are needed when milking is done in automatic milking systems (AMS). Several panel tests were carried out to find out how different consumer groups, milkers and advisors look at and respond to the visual appearance of milk mixed with blood, in order to set a limit for what they think is acceptable. It is concluded from the test panel results that milk samples with 0·4% or more of blood all will be scored as pink and samples with 0·1% blood (about 6 μM-haemoglobin or 100 mg/l) can be visually detected if they are compared with milk samples without blood. The consumer group scored fewer of the samples with 0·1% blood as normal than did the professional groups. The test panel scored 65% of the samples with 1% blood as normal when milk was presented in a black strip cup, which is the reference method when foremilking takes place in a conventional parlour. Only 2% of the milk samples with 2% blood (about 120 μM-haemoglobin or 2000 mg/l) were scored as normal in a black strip cup and should consequently be detected by conventional as well as automatic systems. One model of AMS was tested for its ability to detect and separate milk coloured by blood. The model separated milk with [ges ]6 μM-haemoglobin. Milk mixed with blood injected into the milk stream for a short time at the beginning of milking was not separated. We lack data on how blood is naturally expelled into milk and in what amount. We propose that cow composite milk with >6 μM-haemoglobin should be separated because at this level milk will have a red tinge.

To study the effect of growth hormone (GH) on the functions of mammary epithelia, we examined the effect of GH on the synthesis and secretion of α-casein in a bovine mammary epithelial cell (BMEC) clonal line, which was established from a 26-d-pregnant Holstein heifer. GH receptors (GHR) were observed in the BMEC on the membrane as well as in the cytoplasm. After BMEC were plated onto cell culture inserts, GH stimulated α-casein release in both the presence and absence of the lactogenic hormone complex, which included dexamethasone, insulin and prolactin (DIP). DIP enhanced the effect of GH on α-casein release. Although αs1-casein mRNA expression was not detected in untreated control cells, its expression was observed in BMEC in response to the GH, DIP and GH+DIP treatments. Expression was greater for GH and GH+DIP than for just DIP. Expression of GHR mRNA was increased by DIP treatment, while the mRNA expression was little changed by GH treatment. We conclude that GH acts on BMEC and induces the expression of both the α-casein gene and protein. GHR gene expression was shown to be regulated by DIP and GHR. GHR may be involved in a synergic effect between GH and DIP on casein secretion. These results suggest that GH, in addition to its widely accepted homeorhetic role in vivo, also can act on the mammary parenchyma, and that the effects of GH on mammary epithelial cells could partly account for the clear galactopoietic effect of recombinant bovine GH seen in lactating dairy cows.

An outstanding parameter in cheese making is the type of coagulant, which greatly influences the characteristics of the final products. Proteolysis is the most important set of biochemical changes during ripening of most cheeses, and is carried out, in different magnitude, by proteolytic agents originated in milk, rennet (or rennet substitute), and starter and non-starter micro-organisms (Silva & Malcata, 2000). The demand for alternative sources of milk coagulants, to replace the expensive and limited natural rennet supplies, has increased (Esteves et al. 2001). All commercial enzymes employed as milk coagulant are aspartic proteinases, which are most active at acidic pH and preferentially cleave peptide bonds between residues with hydrophobic side-chains (Silva & Malcata, 1999).

Psychrotrophs, particularly strains of Pseudomonas fluorescens, dominate the microflora of refrigerated raw milk and secrete heat-stable extracellular enzymes (proteases and lipases) which survive pasteurisation and even UHT heat treatments and degrade the casein and fat components of raw milk causing a reduction in cheese yield, gelation of UHT milk and off flavours in many dairy products. These enzymes are usually produced in the late log/early stationary growth phases when the cell density is high. This fact indicated that induction of these enzymes may be a candidate for quorum sensing, a phenomenon by which bacteria can sense and respond to cell population size by means of chemical signals based on the homoserine lactone molecule. It is assumed that the production of these enzymes contributes to the selective advantage of psychrotrophs and hence have a significant effect on their growth kinetics. In the work reported here nine proprietary homoserine lactone compounds were screened, using water as a control, for their effect on the lag phase duration (LPD) and exponential growth rate (EGR) of three strains of Pseudomonas fluorescens isolated from refrigerated raw milk after 1, 3 and 5 days storage. Two compounds viz. N-benzoyloxycarbonyl-L-homoserine lactone and N-3-oxyhexanoyl-DL-homoserine lactone were found to significantly (P<0·001) reduce the LPD and increase the EGR of the three strains. Further work with these compounds is warranted, monitoring growth in parallel with enzyme production, to determine the extent to which extracellular enzyme production is mediated by the quorum sensing phenomenon in this species.

The thermal resistance of the lactococcal bacteriophage P008 was investigated between 55 and 80 °C. Inactivation kinetics revealed an order of reaction above 1 and could be determined by a non-1st-order regression model. Phage inactivation was influenced by the medium (milk and Ca-M17-broth). Within the investigated temperature range, milk had a protective effect on phage P008. This was reflected in the rate constant and in the activation energy. Thermal phage inactivation studies reported in literature were re-analysed using non-1st-order regression. The obtained kinetic parameters showed that phage P008 belongs to the most heat resistant lactococcal phages investigated so far.

Changes in nitrogen compounds during ripening of 40 batches of Manchego cheese made from raw milk (24 batches) or pasteurized milk (16 batches) at five different dairies throughout the year were investigated. After ripening for six months, degradation of p-κ- and β-caseins was more intense in raw milk cheese and degradation of αs2-casein in pasteurized milk cheese. Milk pasteurization had no significant effect on breakdown of αs1-casein. Hydrophobic peptide content did not differ between raw and pasteurized milk cheese, whereas hydrophilic peptide content was higher in raw milk cheese. There were no significant differences between seasons for residual caseins, but hydrophobic peptides were at a higher level in cheese made in autumn and winter and hydrophilic peptides in cheese made in winter and spring. Raw milk cheese had a higher content of total free amino acids and of most individual free amino acids than pasteurized milk cheese. The relative percentages of the individual free amino acids were significantly different for raw milk and pasteurized milk cheeses. The relative percentages of Lys and Ile increased, while those of Val, Leu and Phe decreased during ripening. There were also seasonal variations within the relative percentages of free amino acids. In raw milk cheeses, Asp and Cys were relatively more abundant in those made in autumn, Glu and Arg in cheeses made in winter, and Lys and Ile in cheeses made in spring and summer. Biogenic amines were detected only in raw milk cheese, with the highest levels of histamine, tryptamine and tyramine in cheeses made in spring, winter and spring, respectively.

The objective was to measure teat canal length and diameter, teat diameter and teat wall thickness by ultrasonographic scanning in order to determine the differences in bovine breeds, and to study the influence of teat canal length and diameter on the occurrence of mastitis. A total of 269 lactating dairy cows of four different breeds (Brown Swiss, Simmental, Simmental crossbred with Red Pied, and Holstein-Friesians) from seven Upper Austrian dairy farms were examined. Average teat canal length of Brown Swiss animals was shortest (15·7 mm) followed by Holstein-Friesians (17·2 mm) and Simmental (18·3 mm). These differences in teat canal length were highly significant (P[les ]0·001). There was no significant difference in teat canal length between pure-bred and crossbred Simmentals. Differences of teat canal diameter between breeds were significant (P[les ]0·05). Brown Swiss animals had the largest diameters (2·0 mm) and Holstein-Friesians the smallest (1·7 mm). Differences in teat diameter between Brown Swiss, Holstein-Friesian and Simmental were also significant. No differences were found between the pure-bred and crossbred Simmental cows. The narrowest teats were in Holstein-Friesians and the widest in Simmental. Holstein-Friesians also exhibited the thinnest teat walls while the Simmental had the thickest ones. Teat canal length and diameter were correlated with udder health. Teat canals of healthy udders tended to be longer (17·4 mm) and narrower (1·8 mm) than teat canals of infected udders (15·8 mm, 2·1 mm; P[les ]0·001). A logistic regression model showed significant effects of teat canal length, teat canal diameter and lactation number on udder health.

Dynamics of micro-structural changes in milk during the renneting process were analysed using high-resolution ultrasonic spectroscopy in combination with dynamic rheology and NIR transmission measurements. Two independent ultrasonic parameters, velocity and attenuation were measured in the frequency range 2 to 15 MHz, as a function of time after addition of rennet to milk. The results show an initial decrease of 20 nm for the average diameter of micelles caused by hydrolysis of the κ-casein ‘hairy’ layer followed by an aggregation of the micelles into small clusters (effective aggregation number of 3) and then formation of the gel structure. It was found that evolution of ultrasonic attenuation in the renneting process could well be described by the scattering of the ultrasonic waves on aggregates. The evolution of ultrasonic velocity is well described by the scattering theory but deviates from the predicted curve at the gelation stage of the process, which shows the difference in propagation of ultrasonic waves in a gel structure compared with dispersions. Overall, we found high-resolution ultrasonic spectroscopy to be a powerful tool for analysis of microscopic processes in the formation of milk gel. It allows the characterisation of the pre-gelation processes, such as hydrolysis and aggregation, and the initial stages in the formation of the gel network as well as monitoring of the microscopic evolution in the gel at the post-gelation stage.

A yogurt mix (2 g fat and 17 g solids/100 g) was supplemented with an algae oil emulsion to provide 500 mg ω-3 fatty acids per 272 g serving of yogurt white mass. The emulsion was added to the yogurt mix either before or after the homogenization step and prior to pasteurization. It was then flavoured with a strawberry fruit base and fermented and stored for up to three weeks. The oxidative deterioration of the products was determined by hydroperoxide measurements and by trained and consumer sensory evaluations. The hydroperoxide content of the supplemented yogurts increased over the storage treatment and was unaffected by the stage of addition. The trained panel could distinguish a stronger fishy flavour in both of the supplemented yogurts after 22 days storage, but the consumer panel rated both control and supplemented samples similarly, as ‘moderately liked’.

Genetic polymorphisms in CSN3 gene in Pag (Croatia), Sarda (Italy) and Pramenka (Serbia) sheep breeds were investigated. A single nucleotide polymorphism (SNP) was localized by sequence analysis (sequence submitted to GenBank under accession AY237637) relying on an original primer pair. Primers for sequencing (κ-casF and κ-casR) were designed on the available CSN3 sequences to amplify the genomic region encoding the major part of the mature protein (exon 4). An SNP was detected at position 237 of the sheep κ-casein mRNA (reference sequence: GenBank X51822), where a thymine was substituted for a cytosine. The SNP was typed by conventional PCR and SYBR Green I-based real-time PCR. C and T alleles were discriminated using a dedicated set of primers that consisted of one common forward primer (SNP-TC) and two reverse primers (SNP-T and SNP-C), the latter two differing in the 3′ end base and in the presence of a 12 bp poly-G tail in SNP-C. The SNP was found in both the heterozygous and the homozygous state in Sarda and Pramenka breeds, and in the heterozygous state only in the Pag breed. The observed allelic frequencies of the SNP were 0·12 in Pag, 0·27 in Sarda and 0·45 in Pramenka.

Unidentified constituents in fresh pasture increase milk fat cis-9, trans-11 conjugated linoleic acid (CLA) concentration, and prevent milk fat depression, even though ruminal conditions conducive to reducing milk fat synthesis exist. One possible explanation is vitamin E (α-tocopherol), a constituent high in fresh pasture, but naturally low in conserved/dried forages and cereal grains. Twenty late-lactating dairy cows previously consuming a total mixed ration (TMR) were randomly allocated to one of two dietary treatments for 21 d: TMR (control; n=10); and TMR plus an additional 10000 i.u. α-tocopherol/d (VIT E; n=10). These cows were simultaneously compared with 13 late-lactation dairy cows previously grazing fresh pasture (PAS) balanced for age, parity and genetic merit. Average daily α-tocopherol intakes were approximately 468, 10520 and 1590 i.u./cow for the control, VIT E and PAS treatments, respectively. Dietary α-tocopherol supplementation (VIT E v. control) slightly increased milk fat content by 0·23 percentage units, but did not significantly alter milk fatty acid composition. Plasma trans-11 18[ratio ]1 (VA) content tended to increase and trans-10 18[ratio ]1 levels numerically declined following α-tocopherol supplementation suggesting possible changes in rumen biohydrogenation products. In addition, increased α-tocopherol intake in TMR-fed cows decreased serum urea levels and tended to alter milk fat 15[ratio ]0 suggesting changes in rumen microbial populations. However, when compared with cows grazing pasture, TMR-fed cows supplemented with α-tocopherol, still produced milk with lower cis-9, trans-11 CLA and VA, and higher trans-10 18[ratio ]1 concentrations suggesting α-tocopherol is not a primary reason for milk fatty acid profile differences between pasture and TMR-fed cows. Therefore, additional unknown pasture constituents favour production of fatty acids originating from the cis-9, trans-11 instead of the trans-10, cis-12 CLA biohydrogenation pathways.

Sixty-nine bovine mastitis streptococci belonging to the species Str. agalactiae (n=13), Str. dysgalactiae (n=16), Str. canis (n=22), Str. uberis (n=20) and Str. parauberis (n=4) and six reference strains of the five streptococcal species were examined for their tRNA gene intergenic length polymorphism (tDNA-ILP) fingerprint pattern. Epidemiologically unrelated isolates from bovine mastitis cases were selected by macrorestriction analysis with pulsed-field gel electrophoresis (PFGE). Their results were compared with those obtained from biochemical and serological studies and with those obtained by PCR-mediated identification amplifying species-specific gene segments of the five streptococcal species. According to the present results tDNA-ILP allowed a correct identification of all Str. agalactiae, Str. uberis and Str. parauberis strains investigated also including the reference strains of each species showing species-specific banding pattern. However, all Str. dysgalactiae ssp. dysgalactiae and all Str. canis strains appeared with an undistinguishable pattern which did not allow an identification of the species.

A novel 2-stage gravity separation scheme was developed for fractionation of raw, whole bovine milk into fractions enriched in small (SFG) or large (LFG) fat globules. The volume mean diameter of fat globules in SFG, LFG or control (CTRL) milk was 3·45, 4·68 and 3·58 μm, respectively. The maximum in storage modulus (index of firmness) decreased with increasing fat globule size for rennet-induced gels formed from SFG, LFG or CTRL milks. Miniature (20 g) Cheddar cheeses were manufactured using each of the 3 milks. There were no significant (P>0·05) differences in the pH, moisture and fat in dry matter levels between cheeses made using any of the 3 milks, however, the fat content of the cheese made using SFG milk was ~1% lower than that of cheese made using LFG or CTRL milk in each of the 2 trials. Image analysis of confocal scanning laser micrographs of the cheeses illustrated that the star volume of fat globules in the cheeses decreased significantly (P[les ]0·05) as the size of fat globules in the milks used for cheesemaking was reduced. This indicates that it is possible to manipulate the size distribution of fat globules in Cheddar cheese by adjusting the fat globule size distribution of the milk used for cheesemaking. The concentration of free fatty acids (FFA) increased in all cheeses during ripening. At 120 d of ripening, the concentration of FFA varied significantly (P[les ]0·05 and P[les ]0·001 for trials 1 and 2, respectively) with fat globule size, with cheeses made in trial 2 from LFG, SFG or CTRL milks having total FFA levels of 3391, 2820 and 2612 mg/kg cheese, respectively.

Based on potential health benefits, there is a need to develop effective strategies for enhancing milk fat concentrations of cis-9 18[ratio ]1, 18[ratio ]3 n-3 and conjugated linoleic (CLA) content in milk without compromising the sensory or storage characteristics of processed milk or dairy products. Sixteen Finnish Ayrshire dairy cows were used in a cyclic change-over experiment with four 21-d experimental periods and a 4×2×2 factorial arrangement of treatments to evaluate the effects of forage conservation method, concentrate level and supplements of propylene glycol (PG), and their interactions on milk fatty acid composition and vitamin content. Experimental treatments consisted of four conserved forages offered ad libitum, supplemented with two levels of a standard concentrate (7 or 10 kg/d) and PG (0 and 210 g/d) fed as three equal meals. Primary growths of timothy and meadow fescue sward were conserved by ensiling with none (NA), an inoculant enzyme preparation (IE) or a formic acid based (FORM) additive or as hay 1 week later. Conservation of grass by drying rather than ensiling resulted in lower forage 18[ratio ]2n-6, 18[ratio ]3n-3, total fatty acid and fat-soluble vitamin concentrations. In spite of lower intakes, milk fat 18[ratio ]2n-6 and 18[ratio ]3n-3 content was higher (P<0·05) for hay than for silage diets (12·1, 9·6, 9·6 and 9·3 and 5·00, 3·51, 4·27 and 2·93 g/kg total fatty acids, for hay, NA, IE and FORM silages, respectively). Forage conservation method had no clear effects on milk trans 18[ratio ]1 or CLA content. Compared with silage, hay diets resulted in milk containing lower (P<0·001) riboflavin, α-tocopherol and β-carotene concentrations, but had no effect on ascorbic acid, thiamine, pyridoxine or retinol content. Feeding more concentrates had no effect on milk fatty acid composition or milk vitamin content, other than lowering (P<0·001) 16[ratio ]0 concentrations from 348 to 338 g/kg fatty acids. Supplements of PG led to small (P<0·05) increases in milk 13[ratio ]0 anteiso and 15[ratio ]0 content from 1·06 and 11·3 to 1·22 and 12·6 g/kg fatty acids and reduced (P<0·05) the concentrations of ascorbic acid (16·1 v. 15·1 g/kg milk).

A total of 480 samples of milk from 10 organically and 10 conventionally producing dairy farms in Denmark and covering 8 sampling periods over 1 year (triplicate samplings) were analysed for 45 trace elements and 6 major elements by high-resolution inductively coupled plasma mass spectrometry and inductively coupled plasma atomic emission spectrometry. Sampling, sample preparation, and analysis of the samples were performed under carefully controlled contamination-free conditions. The dairy cattle breeds were Danish-Holstein or Jersey. Sources of variance were quantified, and differences between production systems and breeds were tested. The major source of variation for most elements was week of sampling. Concentrations of Al, Cu, Fe, Mo, Rb, Se, and Zn were within published ranges. Concentrations of As, Cd, Cr, Mn and Pb were lower, and concentrations of Co and Sr were higher than published ranges. Compared with Holsteins, Jerseys produced milk with higher concentrations of Ba, Ca, Cu, Fe, Mg, Mn, Mo, P, Rh, and Zn and with a lower concentration of Bi. The organically produced milk, compared with conventionally produced milk, contained a significantly higher concentration of Mo (48 v. 37 ng/g) and a lower concentration of Ba (43 v. 62 ng/g), Eu (4 v. 7 ng/g), Mn (16 v. 20 ng/g) and Zn (4400 v. 5150 ng/g respectively). The investigation yielded typical concentrations for the following trace elements in milk, for which no or very few data are available: Ba, Bi, Ce, Cs, Eu, Ga, Gd, In, La, Nb, Nd, Pd, Pr, Rh, Sb, Sm, Tb, Te, Th, Ti, Tl, U, V, Y, and Zr.

Microfiltration and ultrafiltration were used to manufacture skim milks with an increased or reduced concentration of whey proteins, while keeping the casein and milk salts concentrations constant. The skim milks were heated on a pilot-scale UHT plant at 80, 90 and 120 °C. The heat-induced denaturation and aggregation of β-lactoglobulin (β-lg), α-lactalbumin (α-la) and bovine serum albumin (BSA) were quantified by polyacrylamide gel electrophoresis. Apparent rate constants and reaction orders were calculated for β-lg, α-la and BSA denaturation. Rates of β-lg, α-la and BSA denaturation increased with increasing whey protein concentration. The rate of α-la and BSA denaturation was affected to a greater extent than β-lg by the change in whey protein concentration. After heating at 120 °C for 160 s, the concentration of β-lg and α-la associated with the casein micelles increased as the initial concentration of whey proteins increased.

We wish to apologise to Vivek Upadhyay for the incorrect spelling of his name on page 226 of volume 72 issue 2 of this journal.

The correct reference is given in full below.

Thom Huppertz, Mathias R Zobrist, Therese Uniacke, Vivek K Upadhyay, Patrick F Fox and Alan L Kelly

Effects of high pressure on some constituents and properties of buffalo milk. Journal of Dairy Research 72 226–233

β-lactoglobulin polymorphisms have been reported in the milk of different goat breeds, although no genetic variants affecting the protein have been characterized. In the present study, we amplified and sequenced the proximal promoter and the first six exons containing the entire coding region for the β-lactoglobulin gene in eleven goat breeds from Spain, France, Italy, Switzerland, Senegal and Asia to identify genetic variants. Fifteen polymorphisms were detected, nine in the promoter region and six in the exons of the β-lactoglobulin gene. All polymorphisms were single nucleotide substitutions with the exception of one deletion/insertion in the promoter region. The polymorphisms in the coding region did not produce any amino acid change. In addition, pyrosequencing technology was used to genotype four polymorphisms in the promoter region in 200 goats belonging to eleven breeds. Differences in allelic frequencies for these polymorphisms between breeds are described and a specific polymorphism for the Italian populations was identified. Finally, the analysis of association between these four promoter point mutations was investigated resulting in five haplotypes, GCGC being the most frequent haplotype in all breeds analysed.