The following compounds were isolated from best-quality Australian butter oil by high vacuum degassing, and identified by the combination of gas chromatography and mass spectrometry: the C3,5,7,9 alkan-2-ones, the C2,4,6,8,10,12 n-alkanoic acids, the C6,8,10 5-lactones (or their hydroxyacids), dimethyl sulphone, all isolated in amounts above 1 pt in 108 of butter oil, and butanone, undecan-2-one, 5-dodecalactone, diacetyl, n-nonanoic acid, a decenoic acid, methyl n-decanoate, toluene, o-methoxyphenol, a cresol, phenol, benzaldehyde, methyl benzoate and benzothiazole, all isolated in amounts between 1 pt in 1011 and 1 pt in 108 of butter oil.

No compound was isolated in an amount greater than flavour threshold, in fact most compounds were below one-tenth threshold level. However, the distillate obtained by degassing produced a very attractive creamy flavour when added to bland non-cultured butter.

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 anti-colon cancer effect of dietary fibre results in part from its fermentation into the short-chain fatty acid butyric acid (BA) by intestinal microflora. BA has potent anti-colon cancer properties owing to its ability to induce apoptosis in colon cancer cells. The colon is not the only location where BA may reach high concentrations, because dietary BA is rapidly absorbed and transported to the liver. We have investigated whether BA could induce apoptosis in transformed human liver (Hep G2) cells. Hep G2 cells treated with BA displayed acetylated histones, increased DNA fragmentation and morphological features consistent with apoptosis. These biochemical features of BA-treated liver cells are identical to those of BA-treated colon cells. In addition, we investigated whether BA present in tributyrin, a triacylglycerol more compatible for inclusion into colloidal lipid structures than BA, could also induce apoptosis in Hep G2 cells. Tributyrin induced DNA fragmentation and morphological features characteristic of apoptotic cells in Hep G2 cells. These results are a significant advance towards delivering BA via colloidal lipid particles to cancerous sites in vivo. This study showed that BA and tributyrin are potent apoptotic agents, and we suggest that sources of dietary BA, such as milk fat, may provide anti-liver cancer properties.

The aim of this study was to examine the effect of bovine colostral whey proteins from cows immunized with Streptococcus mutans/Strep. sobrinus on the adherence and aggregation of caries-inducing bacteria, i.e. mutans streptococci. Both adherence and aggregation are important phenomena in the bacterial colonization of the human oral cavity. In all adherence experiments there was a significant difference between treatments by immune product (IP; from immunized cows) and a control product (CP; a similar product from non-immunized cows). The adherence of 35S-labelled Strep. mutans cells (serotype c) to parotid saliva-coated hydroxyapatite (SHA) was dose-dependently inhibited by both IP and CP if SHA was coated with either product before exposure to bacteria, but markedly lower concentrations of IP than CP were effective. When instead of SHA the bacterial cells were pretreated with IP or CP, only IP strongly and dose-dependently inhibited streptococcal adherence. When bacteria, IP or CP, and SHA were incubated simultaneously, a significant difference between IP and CP treatments was again found. Further, IP effectively aggregated both Strep. mutans and Strep. sobrinus cells, whereas hardly any effect was seen with CP. Both IP and CP aggregated the control bacterium Strep. sanguis, which affected the adherence of the pretreated bacteria.

A method using low resolution NMR spectroscopy is described for investigating whey protein thermal denaturation. The method is based on measuring at 20 °C changes in water proton transverse (T2) relaxation parameter following the denaturing treatment. This parameter is shown to be sensitive to protein denaturation and not to other phenomena such as gelation. Examples are given for the qualitative study of protein thermal denaturation in whey protein concentratc, β-lactoglobulin, α-lactalbumin, bovine serum albumin and immunoglobulins aqueous solutions and for the quantitative determination of thermal denaturation in whey protein concentrate solutions.

Both the rate of formation and the total amount of dimethylglyoxime formed from diacetyl in the presence of hydroxylamine were increased on omission of phosphate from the Owades & Jakovac (1963) hydroxylamine reagent. Sodium acetate was also unnecessary and slightly decreased the rate of dimethylglyoxime formation. The optimum concentration of hydroxylamine lay between 0·5 and 0·75 M. The heating time necessary for formation of dimethylglyoxime can be reduced from 60 to 20min at 75°C. Based on these findings a shortened and more sensitive method for the estimation of diacetyl is recommended.

Part I

1. A method for the estimation of vitamin B1 in milk by a modified Jansen test involving takaphosphatase treatment is described in detail.

Part II

2. It is shown by ultra-filtration experiments, by treatment with pepsin and takaphosphatase, with trichloroacetic acid and with fuller's earth that vitamin Bx is present in milk unesterified and phosphorylated and that both forms may be freely diffusible or combined with protein either loosely or by firmer bonds.

Part III

3. The effect of the stage of lactation on the amounts and partition of vitamin B1 in milk has been studied. Samples of colostrum and early milk were found to contain large amounts of cocarboxylase (up to 80 % of the total). Hardly any cocarboxylase was present in mid or late lactation milks. The total vitamin B1 content was 60–100μg./100 ml. for colostrum, up to 60μg./100 ml. for early milk and 30–40μg./100 ml. for mid and late lactation.

4. A marked negative correlation exists between the cocarboxylase content of cow's and goat's milk and their alkaline phosphomonoesterase titre. As this enzyme cannot dephosphorylate cocarboxylase it is probably accompanied in milk by other enzymes more suited to this process.

5. The suggestion is put forward that the vitamin B1 of milk is mostly derived not directly from the blood but from the secretory cells of the mammary gland.

In the course of routine bacteriological examinations of milk carried out at one of the United Dairies pasteurising depots during the past three years, a remarkable lack of constant relationship between the numbers of colonies reported on the 1/10, 1/100 and 1/1000 dilution plates, when standard agar (the Ministry of Health formula) was used, has been observed. When, however, standard agar, enriched with 0·5 per cent, milk, has been used as a plating medium, many of the discrepancies between different dilutions of the same milk have disappeared.

A method is described for preparing immunologically homogeneous human milk β-casein, against which monospecific rabbit antiserum was prepared. The antiserum was used to quantify β-casein, the major human casein, by rocket Immunoelectrophoresis in individual milk samples. However, it was found that in most samples β-casein occurred together with degradation products originating from its proteolysis by plasmin. Immunological quantification of human β-casein, treated with plasmin for various time periods, showed that rocket height was not affected by proteolysis up to degradation states clearly more advanced than those observed in all samples of fresh human milk tested. Assays of 150 individual milk samples from 80 women, covering a lactation period of up to 730 d, gave an average concentration of β-casein (native+degraded) of 4·67±0·89 standard deviation (g/1); extremes at 2·1 and 7·3 g/1 did not vary significantly during the period under study. Comparison of this average value with an accepted casein content of 4·4 g/1 (Macy & Kelly, 1961) showed that the casein content of human milk is underestimated when obtained by N determinations on milk and on its supernatants at pH 4·6 (whey). Caseins other than β-casein occurred only in minute amounts, if at all.

Fifty-seven inoculations of a strain of Staphylococcus aureus were made into the outer 4 mm of the teat ducts of 8 cows. The inocula ranged from 10 to 600 colony-forming units (CFU) and were made with a stainless steel instrument designed to overcome the disadvantages of glass rods and cotton swabs hitherto used.

A single inoculation resulted in either no colonization of the duct, in temporary colonization for up to 7 days, or in colonization followed by intramammary infection.

The recovery of the organisms depended on the size of the inoculum. When 70–100 CFU were placed in the teat duct no organisms were recovered from 12 of 24 quarters after the 1st post-inoculation milking. When 500–600 CFU were used, organisms were recovered from all of 31 quarters for at least 3 milkings, and from 61% for 6 milkings or more, in spite of dipping the teats in a strong disinfectant twice daily.

Intramammary infection developed in 1 of 12 quarters (8%) inoculated in the teat duct with about 600 CFU when the animals were milked twice daily, and in 5 of 19 (23%) quarters if the 1st post-inoculation milking was omitted.

There was no evidence of sensitization resulting from previous infection.

Based on a previous ensiling study in glass silos of various pomegranate pulp (PP) mixtures, fresh pomegranate pulp (PP) was mixed with drier feeds including soy hulls and corn silage (40:35:25 on DM basis) and ensiled in 32 pressed bales (700 kg each) wrapped with stretch polyethylene film. This ensiled pomegranate pulp mixture (PPM) was included in lactating cow total mixed ration (TMR) at a level of 20% of DM (PPM-TMR). Performance and digestion experiment was conducted with two groups of 21 milking cows each, fed individually one of the two TMR: 1. Control TMR without ensiled PPM; 2. Experimental TMR which contained 20% ensiled PPM, including 8% PP as corn grain replacer. Voluntary DM intake of cows fed the control TMR was 5·04% higher than that of the PPM cows. In vivo digestibility of DM, OM, NDF, CP and fat were significantly higher in the control cows compared with the PPM group, but methane production in the rumen fluid was 25% lower in the PPM cows. A slightly higher milk yield (by 2·2%) observed in the control cows; however, milk fat content was 5·9% higher in the PPM cows. This was reflected in similar yield of energy corrected milk (ECM) and 3·97% increase in production efficiency (ECM/DM intake) of the PPM cows compared with the control ones. Welfare of the cows, as assessed by length of daily recumbence time, was in the normal range for both groups. Body weight gain was also similar in both groups. Data suggest that the level of 8% PP in the PPM-TMR used in this study was probably too high for lactating cows and should be lowered to 4% in order to achieve better performance.

β-Galactosidases from Kluyveromyces lactis and Kluyveromyces marxianus isolated from artisanal ewes’ milk cheeses, were used to transgalactosylate lactose from cheese whey permeate (WP). The content of galactooligosaccharides (GOS) obtained by transgalactosylation was comparable with that formed using pure lactose as substrate. In order to obtain a mixture with higher prebiotic oligosaccharide content, isomerisation of the transgalactosylated WP was carried out using sodium aluminate as catalyst. The transgalactosylated mixtures at 6 h of reaction contained amounts of prebiotic carbohydrates (tagatose, lactulose, GOS and oligosaccharides derived from lactulose, OsLu) close to 50 g/100 g of total carbohydrates for all the strains tested, corresponding to 322 g prebiotics/kg whey permeate. Thus, the suitability of this methodology to produce mixtures of dietary non-digestible carbohydrates with prebiotic properties from WP has been demonstrated, which is interesting for the food industry since it increases the value and the applicability of this by-product from cheese manufacture.

The imprinted insulin-like growth factor 2 gene (IGF2) encodes a fetal mitogenic hormone protein (IGF-II) and has previously been shown to be associated with performance in dairy cattle. In this study we assessed genotype-phenotype associations between four single nucleotide polymorphisms (SNPs) located within the bovine IGF2 locus on chromosome 29 and a range of performance traits related to milk production, animal growth and body size, fertility and progeny survival in 848 progeny-tested Irish Holstein-Friesian sires. Two of the four SNPs (rs42196909 and IGF2.g-3815A>G), which were in strong linkage disequilibrium (r2=0·995), were associated with milk yield (P⩽0·01) and milk protein yield (P⩽0·05); the rs42196901 SNP was also associated (P⩽0·05) with milk fat yield. Associations (P⩽0·05) with milk fat percentage and milk protein percentage were observed at the rs42196901 and IGF2.g-3815A>G SNPs, respectively. The rs42196909 and IGF2.g-3815A>G SNPs were also associated with progeny carcass conformation (P⩽0·05), while an association (P⩽0·01) with progeny carcass weight was observed at the rs42194733 SNP locus. None of the four SNPs were associated with body size, fertility and progeny survival. These findings support previous work which suggests that the IGF2 locus is an important biological regulator of milk production in dairy cattle and add to an accumulating body of research showing that imprinted genes influence many complex performance traits in cattle.

Bacteriological analyses, cell counts using the Fossomatic method and California Mastitis Test were performed on 1523 goat milk samples taken aseptically at monthly intervals throughout lactation from three goat herds. Of the goat udders, 81·4% were infected, minor pathogens being the most frequent isolates (65·7%). Differences in the level of infection by minor pathogens were found betwccn herds. Cell counts were influenced by stage of lactation and intramammary infection. Cell counts < 106 cells/ml were found in 80% of milk samples infected by major pathogens and in 45% infected by minor pathogens. About 81% of udders infected with major pathogens gave California Mastitis Test scores of 2 and 3, compared with 20% for uninfected goats. A high proportion (65%) of udders infected with minor pathogens also produced scores of 2 and 3. A significant positive correlation was found between the California Mastitis Test and cell counts. The use of cell counts for the detection of abnormal goat milk is discussed.

1. The mixed evening and morning milk from 360 cows, all sampled six times during their lactation, was analysed chemically and for its cell content. A total and a differential cell count were made. Similar analyses were made on 823 samples of farm milk taken at a receiving creamery.

2. The total cell counts varied from less than 10,000 to 5,000,000/ml. and there was considerable variation in the chemical composition of samples containing similar numbers and types of cells.

3. The proportion of polymorph cells in the total count increased rapidly with increasing total count.

4. Stage of lactation had little effect on the number of cells present in the milk, but samples from cows in very early and very late lactation had been avoided. The total cell count was lowest from the 70th to 130th day of lactation with a tendency to rise after the 130th day.

5. Both the total and the polymorph cell counts of the milk increased sharply with the lactation number of the cow until the seventh lactation and then remained at a fairly constant level.

6. As the total cell count increased to an average value of 500,000/ml. there was a fall in the daily yield of milk per cow and a marked fall (0·25%) in solids-not-fat content. The latter was the result of a steady drop in the lactose content, the casein content being affected only by counts approaching 1,000,000/ml. An increasing cell count was without effect on the fat content. Possible reasons for the relatively small effect on chemical composition of high cell counts in some milks is discussed.

7. It is concluded that the presence of subclinical mastitis, as evidenced by the cell count of the milk, leads to a lowering of the solids-not-fat content.

The authors wish to thank their assistants in the Technical Chemistry and Veterinary Pathology Departments, who did most of the practical work.

A method is described for the chromatographic analysis of the headspace of hard cheese. Among the cheeses examined were 12 commercial cheeses and 12 Cheddars made in the NIRD experimental dairy. The 12 commercial cheeses were Parmesan, Caerphilly, Stilton, Leicester, Double Gloucester, Cheshire, Gouda, Wensleydale and 4 varieties of Cheddar. It was found that those varieties containing low concentrations of sulphur compounds had little aroma and derived most of their flavour from taste and texture. The concentration of 2-pentanone was found to be a good indicator of maturity of Cheddar cheese.

Aminopeptidase P was purified 65·3-fold from the cytoplasm of Lactococcus lactis subsp. cremoris AM2 with a 5·8% yield. The purified enzyme was found to consist of one polypeptide chain with a relative molecular mass of 41600. Metal chelating agents were found to be inhibitory and Mn2+ and Co2+ stimulated activity 7-fold and 6-fold respectively. The purified enzyme removed the N-terminal amino acid from peptides only where proline (and in one case alanine) was present in the penultimate position. No hydrolysis was observed either with dipeptides even when proline was present in the C-terminal position or when either N-terminal proline or pyroglutamate was present preceding a proline residue in the penultimate position of longer peptides. On the basis of this substrate specificity either aminopeptidase P or post-proline dipeptidyl aminopeptidase are necessary along with a broad specificity aminopeptidase to effect complete hydrolysis of casein-derived peptides containing a single internally placed proline residue. However, both aminopeptidase P and post-proline dipeptidyl aminopeptidase would be required together with a broad specificity aminopeptidase in order to completely hydrolyse casein-derived peptides that contain two internally placed consecutive proline residues. As bitter casein-derived peptides are likely to contain either single prolines or pairs of prolines, aminopeptidase P appears to be an important enzyme for debittering.

A method for the estimation of urea in milk using ammonia electrode is described. Urea is first degraded by urease enzyme into ammonium ion and carbon dioxide at neutral pH. The ammonium ion is then converted into ammonia at alkaline pH. A linear inverse relationship was observed between logarithmic concentration of ammonia or urea and electrode response. Repeatability, expressed as a coefficient of variation, was 1·77% at a level of 8·92 mm-urea in milk. The method was validated in milk samples spiked with between 2×10−3 and 10×10−3 m-urea and recovery of added urea was quantitative. Whereas, preservative sodium azide at 0·5 g/l or 2 g/l level did not affect results, lower values of urea concentration in presence of Bronopol at 0·5 g/l were observed. Urea levels in milk samples estimated by this method were comparable to standard enzymatic method. The method is simple, fast and is not prone to interference from other milk constituents.

The variations in the composition of milk over an 18-week period from lactating British Alpine and Anglo–Nubian goats imported into Trinidad are reported. The animals were fed Napier grass (Pennisetum purpureum) ad lib. and 2 lb of a proprietary concentrate mixture daily.

The mean percentage contents of the milks of British Alpine and Anglo–Nubian goats were: total solids 11·49±0·22, 12·17±0·28; butterfat 3·42±0·18, 4·06±0·22; total N 0·462±0·04, 0·530±0·07; ash 0·78±0·02, 0·79±0·03 and ‘lactose’ 4·38±0·28, 4·05±0·55. Anglo–Nubian goats had a higher content of percentage total solids, fat, total N and ash. For all constituents the coefficient of variation between weeks was higher for Anglo–Nubian goats. No significant trends were found between weeks for each of the components analysed for both breeds. Statistically significant correlations (P < 0·01) were found between percentage fat and percentage total N for both breeds.

The mean mineral contents (mg/100ml) for the milks of British Alpine and Anglo–Nubian goats were: Na 52·6±4·6, 55·5±1·9; K164·0±8·4, 133·4±11·4; Ca 85·5±11·6, 89·9±9·8; Mg 13·2±3·8, 10·2±1·6 and P 74·7±3·9, 82·1±3·9. British Alpine goat's milk had a higher content of K and that of Anglo–Nubian goats had a higher content of P; both differences were statistically significant (P < 0·01).

The milk composition figures in this study together with those from a similar study in Guadeloupe suggest that the values are low and considerably lower in comparison with the same breeds of goats in a temperate environment. This could be a feature of dairy goats imported into the tropics due perhaps to inadequate nutrition. Much more information is needed to verify this possibility.

This study was designed to assess anti-diabetic potential of goat, camel, cow and buffalo milk in streptozotocin (STZ) induced type 1 diabetic albino wistar rats. A total of 48 rats were taken for the study where one group was kept as non-diabetic control group (8 rats) while others (40 rats) were made diabetic by STZ (50 mg/kg of body weight) injection. Among diabetic rats, a control group (8 rats) was kept and referred as diabetic control whereas other four groups (8 rats each) of diabetic rats were fed on 50 ml of goat or camel or cow or buffalo milk for 4 weeks. All the rats (non-diabetic and diabetic) were maintained on standard diet for four weeks. STZ administration resulted in enhancement of glucose, total cholesterol, triglyceride, low density lipoprotein, HbA1c and reduction in high density lipoprotein in plasma and lowering of antioxidative enzymes (catalase, glutathione peroxidase and superoxide dismutase) activities in pancreas, kidney, liver and RBCs, coupled with enhanced levels of TBARS and protein carbonyls in pancreas, kidney, liver and plasma. OGTT carried out at the end of 4 week milk feeding indicated that all milks helped in early maintenance of glucose level. All milks reduced atherogenic index. In camel milk fed diabetic group, insulin concentration enhanced to level noted for non-diabetic control while goat, cow and buffalo milk failed to restore insulin level. HbA1c level was also restored only in camel milk fed diabetic group. The level of antioxidative enzymes (catalase, GPx and SOD) in pancreas enhanced in all milk fed groups. Camel milk and to a reasonable extent goat milk reduced formation of TBARS and PCs in tissues and blood. It can be concluded that camel milk ameliorates hyperglycaemia and oxidative damage in type-1 diabetic experimental rats. Further, only camel milk completely ameliorated oxidative damage in pancreas and normalised insulin level.