A new method is described for the determination of the free and masked sulphydryl (-SH) groups in heated milk and milk products. The procedure is based on the use of the specific -SH reagent p-chloromercuribenzoate, and utilizes another specific -SH reagent, 5,5' -dithiobis(2-nitrobenzoate), as an indicator producing yellow colours which are matched with the aid of standard glasses in the Lovibond Comparator.

The method was applied to heated skim-milks and a series of commercially prepared milk powders. Measurements were made in particular of the -SH content of UHT-treated milk. The changes in the -SH content of such milks on storage were studied; these changes could not be explained by a reversal of the denaturation of the whey proteins, although evidence was obtained that small amounts of heatdenatured whey protein can revert to the native state.

Two monoclonal antibodies (MAb) raised against bovine κ-casein were developed and applied in an automated optical biosensor (Biacore 3000) to create easy and fast direct and inhibition biosensor immunoassays (BIA) for the detection of cows' milk in the milk of ewes and goats. With both assay formats, low limits of detection (<0·1%) and fast run times (around 5 min) were obtained. For sample preparation, milk was diluted in buffer (direct assay) or in an antibody-containing buffer (inhibition assay) only. For quantitative analysis, calibrants of cows' milk in ewes' or goats' milk were used. Advantages of the direct BIA are: the single reagent format (biosensor chip immobilized antibodies only); the use of small amounts of antibodies (2 μg for >350 tests); and the wide measurement range (0·1 to 10% cows' milk). Despite these advantages, the inhibition BIA (using κ-casein immobilized on the chip) was preferred because of the possible application of non-purified Mab, the higher responses, the higher sensitivity at relevant low percentages of cows' milk and its robustness (>800 cycles per chip).

Of the lactic acid bacteria isolated, 720 strains from thirty-six typical Cheddar cheeses have been classified. The dominant organism in New Zealand cheese has been shown to be Sbm. plantarum. Sbm. casei occurs much less frequently, while betabacteria and betacocci are found in still smaller numbers. Betabacteria tend to replace Sbm. plantarum in the final stage of cheeseripening. The flora of good quality cheese has been found to consist mainly of one or two varieties of Sbm. plantarum, often associated with Sbm. casei or a small proportion of betabacteria. In poor-quality cheese a larger number of types of lactic acid bacteria frequently occur. A simple routine method is described for distinguishing the different groups present in cheese.

The effects which different strains of lactobacilli and betacocci produce in cheese have been correlated with their biochemical characteristics. Sbm. casei in general appears to be beneficial to cheese quality. The strains of Sbm. plantarum isolated have been grouped in four varieties, mainly on the basis of sugar reactions; one variety has been shown to be beneficial to cheese quality, another variety has very little apparent effect, while the remaining two varieties produce serious defects—bad flavours, discoloration and, occasionally, open texture. The same defects are produced by most strains of betabacteria and betacocci when present in large numbers.

The biochemical characteristics of the lactobacilli studied have shown a more regular gradation from Sbm. casei through Sbm. plantarum to Bbm. breve (the only species of Betabacterium isolated) than is indicated from the generally accepted systems of classification.

A proteinase was purified from a cell wall extract of a culture of Streptococcus lactis NCDO 763 grown in skim milk. Being active at a low pH (at pH 4·8 on haemoglobin and pH 6·–6·5 on casein) and completely inhibited by diisopropylfluorophosphate, it was considered to be a serine proteinase partly inhibited by EDTA; the mol. wt was ∼ 80000.

Machine milking induced repeatable, short-term changes in thickness of the teat apex. These changes were measured with a spring-loaded caliper instrument under milking conditions that imposed differing forces on the teat apex. The mean percentage changes in teat end thickness (relative to the premilking values for individual teats) varied from 10% decrease up to 20% or more increase depending on the particular milking system used. The overall decrease in teat end thickness associated with cannula milking, milking at very low vacuum levels or milking with positive pressure pulsation may result from: (i) decrease in the intramammary pressure; and/or (ii) small changes in tone of teat musculature; and/or (iii) changes in the distribution of interstitial fluids in the teat apex. The overall increase in teat thickness caused by conventional or PKME milking systems results mainly from oedema (the extravascular accumulation of fluid). It is likely that milking equipment can, and should, be designed to minimize circulatory impairment resulting in oedema of the teat.

Cows were fed either 75 or 100% of the recommended intake levels for protein and 100% of recommended energy levels (Agricultural Research Council, 1965) from 8 weeks pre-calving until 14 weeks post calving. From 14 weeks post calving and to the end of lactation all the cows received 100% of the recommended protein and energy intakes.

The mean of the 305-d milk yields of the 2 groups was not significantly different and although cows on the lower protein intake produced less lactose during the first 14 weeks of lactation there was no significant difference in total lactose, fat, protein or total solids production between the groups. In both groups blood packed-cell volume, red cell count and haemoglobin decreased during the first 10 weeks of lactation and then began to increase in the high-protein group. The cows receiving the low-protein diet showed a similar increase only when they received the high-protein ration from 14 weeks post calving. The mean interval from calving to conception was 27·5 weeks in the high-protein group and 20 weeks in the low-protein group.

It is concluded that feeding 75% of protein requirements to dairy cows during the first 14 weeks of lactation does not reduce milk yield or quality significantly and probably has no adverse effect on fertility.

Soluble proteins and Ca and P salts in the fouling deposit on a mineral microfiltration membrane obtained in static conditions were analysed and compared with those found using dynamic filter conditions in a laboratory pilot installation for milk processing. The serum milk proteins were retained by the porous micellar deposit under both static and dynamic conditions, resulting in progressive fouling of the dynamic micellar membrane, enabling milk to be processed with a mineral microfiltration membrane. Ca and P salts increased fouling probably by allowing better adsorption of casein micelles on the alumina and by acting as intermicellar bonds in the deposit. Fat globules modified porosity, permeability and resistance to matter transfer of the deposit. The dynamic conditions gave a deposit of the same type and structure as with the static conditions although made thinner by the velocity effect.

Machine-induced changes in teat thickness were measured in a randomly selected group of 22 cows from a commercial dairy herd consisting of 110 cows during an experiment lasting 1 year. Half the cows used were free from mastitis whereas the remainder had at least one quarter infected by environmental pathogens. Teats were classified according to a threshold change in teat end thickness of 5%. The relationships between quarter infection or teat duct colonization and teat end thickness changes induced by machine milking were investigated. Teats with > 5% change in thickness had significantly increased teat duct colonization and a slight, non-significant, increase in quarter infection. We conclude that machine-induced changes in teat end thickness are predisposing factors for teat duct colonization by environmental pathogens. This may provid: an increased new infection risk, especially when hygiene is poor.

Starting in week 5 of lactation, goats were milked unilaterally once daily for 4 ½ weeks, then twice daily for 12 weeks and finally thrice daily for 2 weeks. The control gland was milked twice daily throughout. During less frequent milking, yield fell by 26%. There was a rapid and essentially complete recovery of milk yield on resumption of twice-daily milking, and five of the six goats subsequently responded to thrice-daily milking with a small increase in milk yield. Mammary function was assessed during each phase by determining enzyme activities and in vitro synthesis rates of lactose and casein in mammary biopsies. There was a trend towards reduced differentiation of secretory cells during infrequent milking, with very little recovery during twice-daily milking but complete recovery during thrice-daily milking. The differentiative state of the control gland remained unchanged throughout.

Preparation, physicochemical characterization and stability of iron-supplemented caseins were studied. When ferrous iron (final concentrations of 0·25, 0·5, 1·0 and 1·5 mM) was added to sodium caseinate (final casein concentration 25 g/l), iron binding to caseins was complete. In parallel, an increase in absorbance at 280 nm, a decrease in fluorescence intensity, modifications of reversed-phase HPLC profile, a decrease in pH and an increase in free sodium concentration were observed. Moreover, no release of iron from iron-casein complexes was found between pH 6·5 and 3·7 or after heat treatments or in the presence of inorganic phosphate. Only EDTA disodium salt and trisodium citrate had significant effects on the release of iron bound to caseins.

Constituents of a flavour concentrate from fishy butterfat were separated by gas chromatography on silicone oil and ‘Carbowax 400’ columns. Two fractions with distinct flavours were obtained, the one of oily flavour and containing n-hexanal, n-heptanal, hex-2-enal and heptan-2-one, the other of metallic flavour, containing a single carbonyl compound present in relatively small amount. It is considered that these compounds with the exception of heptan-2-one are mainly responsible for the characteristic fish-oil flavour.

Other compounds isolated include propanal, n-pentanal, n-octanal, n-nonanal, n-decanal, acetone, pentan-2-one, nonan-2-one, undecan-2-one, acraldehyde, pent-2-enal, hept-2-enal, oct-2-enal, non-2-enal, hepta-2, 4-dienal, a compound resembling octa-2, 4-dienal, a compound with a mushroom flavour, and a compound that formed the 2, 4-dinitrophenylosazone of methylglyoxal.

Indirectly heated ultra-high-temperature (UHT) processed milk was prepared with initially high, medium, and low dissolved O2 contents of 8·9, 3·6 and 1·0 ppm respectively, aseptically bottled, and tested at intervals during storage at room temperature for 150 d. Flavour acceptability increased to a maximum after a few days, but declined slowly after about 6 d; the increase was associated with less off-flavour described as ‘cabbagey’, and the decrease with more ‘stale’ off-flavour descriptions. Milks with higher initial O2 contents were preferred up to 8–13 d, but thereafter acceptability was independent of initial O2 content. Sulphydryl group (–SH) contents rapidly decreased and O2 levels correspondingly declined in the first few days as the flavour improved. Loss of –SH was lower with lower initial O2 contents, and moderate –SH content remained in low O2 samples for several weeks. Ferricyanide reducing (FR) values did not satisfactorily measure stale flavour development. They were initially high and decreased during the first 13 d at rates dependent on O2 content. After 20 d the FR values began to rise in high O2 samples, but continued to decline slowly in low O2 samples up to 90d although stale flavour was increasing.

High initial O2 contents resulted in rapid depletion of ascorbic acid and folic acid during storage. Losses of vitamin B12 were small, but were higher with high O2 contents than with low.

The beneficial effect of O2 on flavour, therefore, appears to be so slight and confined to such a short period in the early life of the milk as to be completely outweighed by the adverse nutritional effects.

The 31P nuclear magnetic resonance spectrum of liquid milk was examined. Of the three peaks observed, the two larger were assigned to inorganic phosphate (Pi) and the seryl phosphate (SerP) residues of casein; the third peak was assigned to a phosphodiester, which is probably glycerophosphoryl choline. The pH-dependences of the chemical shifts of the Pi and SerP were measured with and without added EDTA, and the results confirm the assignments. The width of the Pi peak in milk is significantly greater than in similar solutions lacking casein, probably because of binding to, or chemical exchange with, the casein micelle. Most of the SerP residues in milk are not sufficiently mobile to have been detected in these experiments but a significant fraction of SerP residues is able to move freely and can be titrated.

This study investigated the application of near infrared (NIR) reflectance spectroscopy to the measurement of texture (sensory and instrumental) in experimental processed cheese samples. Spectra (750 to 2498 nm) of cheeses were recorded after 2 and 4 weeks storage at 4 °C. Trained assessors evaluated 9 sensory properties, a texture profile analyser (TPA) was used to record 5 instrumental parameters and cheese ‘meltability’ was measured by computer vision. Predictive models for sensory and instrumental texture parameters were developed using partial least squares regression on raw or pre-treated spectral data. Sensory attributes and instrumental texture measurements were modelled with sufficient accuracy to recommend the use of NIR reflectance spectroscopy for routine quality assessment of processed cheese.

Late blowing, caused by the outgrowth of clostridial spores present in raw milk and originating from silage, can create considerable product loss, especially in the production of hard and semi-hard cheeses. The conventional method for the isolation of Clostridium spp. from cheeses with late-blowing symptoms is very complicated and the identification of isolates is problematic. The aim of this work was the development of a multiplex PCR method for the detection of the main dairy-related clostridia such as: Cl. beijerinckii, Cl. butyricum, Cl. sporogenes, Cl. tyrobutyricum. Samples derived from silage, raw milk and hard cheese were analysed by the most probable number (MPN) enumeration. Forty-four bacterial strains isolated from gas positive tubes were used to check the reliability of the multiplex PCR assay. The specificity of the primers was tested by individually analysing each primer pair and the primer pair combined in the multiplex PCR. It was interesting to note that the samples not identified by the multiplex PCR assay were amplified by V2–V3 16S rRNA primer pair and the sequencing revealed the aligned 16S rRNA sequences to be Paenibacillus and Bacillus spp. This new molecular assay provides a simple promising alternative to traditional microbiological methods for a rapid, sensitive detection of clostridia in dairy products.

Colostrum and milk samples from 20 goats were analysed for concentrations of immunoglobulin G (IgG), β-lactoglobulin (β-lg), α-lactalbumin (α-la) and serum albumin (CSA) throughout the first 14 milkings post partum (7 d of lactation) using single radial immunodiffusion assay. Concentrations (mg/ml, means±SD) at first milking were IgG 47·9±25·5, β-lg 30·7±10·4, α-la 2·77±0·82 and CSA 2·97±2·46 mg/ml. Large variations were recorded for IgG concentrations (19·9–94·5 mg/ml) and β-lg (9·3–49·8 mg/ml). Concentrations of IgG, β-lg and CSA dropped abruptly in the subsequent milkings and α-la concentration decreased slowly. Mean IgG concentration was <2 mg/ml after 7 milkings and <1 mg/ml after 11 milkings. However, IgG concentration does not differ significantly, at the 1% level, from milkings 7–14. The contribution of β-lg to the increase in whey proteins in early milks was greater than that of IgG from milkings 5 to 14. The results were tabulated to make it possible to calculate the excess of whey proteins that would be obtained if early milks were illegally added to milk supply.

The addition of simple aldehydes brought about large increases in the heat stability of both skim-milk and concentrated skim-milk over a comparatively wide milk–pH range. The coagulation time–pH minima of type A milks were completely removed by aldehyde treatment. Some sugars, which react readily as aldehydes on heating, were also shown to stabilize concentrated milk to prolonged heat treatment at 120 °C.

Sow milk protects the piglet against infectious diseases through a variety of mechanisms. In this study, the presence of potentially probiotic lactic acid bacteria in this biological fluid was investigated. Milk samples were obtained from 8 sows and a total of 19 rod-shaped isolates were selected for identification and assessment of their probiotic potential. RAPD profiling revealed the existence of 8 different genetic profiles among them. One representative of each profile was selected for further characterization and they were identified as Lactobacillus reuteri, Lb. salivarius, Lb. plantarum, Lb. paraplantarum, Lb. brevis and Weissella paramesenteroides. Then, their probiotic potential was evaluated through different assays, including survival in conditions simulating those existing in the gastrointestinal tract, production of antimicrobial compounds, adherence to intestinal mucin, production of biogenic amines, degradation of mucin, and pattern of antibiotic sensitivity. Three strains, Lb. reuteri CR20 (a reuterin-producing strain), Lb. salivarius CELA2 (a bacteriocin-producing strain) and Lb. paraplantarum CLB7 displayed the highest probiotic potential.

An analytical procedure, based upon HPLC, has been used to determine the degree of isomerization of retinol (vitamin A) in various dairy products. In raw milks, which are not subjected to thermal processing, there was no conversion of the predominant all-trans-isomers to cis-isomers in samples from various species. Pasteurized milks with mild heat treatment (high quality milk) had an average 13-cis[ratio ]all-trans ratio of 2·6%, while pasteurized milk treated for 15 s at temperatures ranging from 72 to 76°C had an average ratio of 6·4%. Milk subjected to more severe heat treatments had a higher degree of isomerization (UHT milk, 15·7%; sterilized milk, 33·5%), consistent with increased thermal conversion of the retinol isomers. In pasteurized and UHT cream, the increase in 13-cis-isomer was also a consequence of heat treatment (pasteurized cream, 3·0%; UHT cream, 14·4%). The presence of cis isomers in fermented milk suggests that fermentation processes, directly or indirectly, can induce cis–trans isomerization. In the cheeses analysed, the extent of retinol isomerization ranged from 7·6 to 35·0%. Our results confirm the importance of individual isomers in evaluating the vitamin A status of dairy products.