The effects on lactation of perphenazine, a tranquillizer which increases the level of prolactin in the blood, have been studied in cows, goats, guinea-pigs and rabbits. The drug caused a significant depression of milk yield in cows, goats and rabbits, but was without effect on the milk yield of guinea-pigs. It seems probable that the inhibitory effects on lactation were due to the actions of the drug on appetite and behaviour. It is concluded that perphenazine, because of its other actions, is not a suitable agent for studying the physiological effects of increased levels of endogenous prolactin.

Spontaneous rhythmical contractions of the teats of living cows were recorded by a plethysmographical method. These contractions were inhibited by isoproterenol, adrenaline, isoxsuprine and phentolamine, but were activated by phenylephrine. Noradrenaline induced activation which was often followed by inhibition.

These results obtained in vivo were in agreement with responses obtained in vitro. Apparently beta sympathomimetic effects are evoked easily under in vivo conditions.

Milks (bovine and human) and dairy products (butter, cheese, skim and whey powders, calf-replacer, casein, butter-oil and dietetic food) were collected during 1971/2 throughout Ireland together with a more limited sample of the 10 major animal feed ingredients, and analysed for organochlorine insecticide residues using electron-capture gas chromatography. The different materials contained low or negligible levels of chlorinated insecticides. Apart from some of the animal feed ingredients the DDT residues were generally the predominant contaminants detected together with lower levels of γ-BHC (lindane), aldrin/dieldrin and heptachlor/heptachlor epoxide. The maximum levels of these insecticides in the bovine milk and dairy products (511, 100, 62 and 21 μg/kg fat respectively) constitute only 50% or less of the Codex Tolerance Limits. The correspondingly low residue levels in the human milk (maxima of 128, 1, 1, and 5 μg/kg fat respectively) which at most represent insecticidal ingestion by infants equivalent to 13, 0·05, 5 and 5% respectively of the WHO/FAO acceptable daily intake for DDT, γ-BHC, aldrin/dieldrin and heptachlor/heptachlor epoxide again pose no obvious health hazards and are strongly indicative of negligible organochlorine contamination in the general diet. The samples of animal feed ingredients examined also contained trace levels of organochlorines (maxima of 0·9, 0·1, 1·6 and 1·0 μg/kg respectively). More extensive monitoring of the residues in animal feed ingredients (the most probable source of milk contamination) is advocated, and the desirability of tolerance limits for insecticides in animal feeds discussed.

Raw bulk milk samples were stored at 5°C in the laboratory mostly for 7 or 8d and examined at intervals for bacterial content, proteolysis as indicated by tyrosine value, and organoleptic quality. Data are presented for these samples illustrating relationships between bacterial population counts, tyrosine values and organoleptic quality ratings. Bacterial counts (total bacteria, psychrotrophs and proteolytic psychrotrophs) generally exceeded 106/ml before a definite increase in tyrosine value or any deterioration in organoleptic quality could be detected. Tyrosine value was indicated to be useful as a general index of bacteriological and organoleptic quality of commercial raw milk subjected to prolonged cold storage. The tyrosine value procedure was also shown to have application for monitoring the development of proteolysis in cold-stored pasteurized milk and cream.

The principal results of an investigation of the major physico-chemical properties of porcine milk were:

Heat stability: all individual porcine milks examined had poor heat stability and some coagulated in 2 min at 95°C; the Q10°C was high and variable and was 40 for some samples. Dialysis of porcine milk against bulk bovine milk did not influence its heat stability; removal of colloidal calcium phosphate (CCP) from porcine milk increased its heat stability 4-fold, but it was still very unstable. Unlike bovine milk, porcine milk did not exhibit a maximum-minimum in the heat stability-pH curve, but rather resembled bovine casein micelles suspended in milk dialysate. Addition of isolated bovine β-lactoglobulin to porcine milk did not influence its heat stability-pH curve.

Rennet coagulation: under similar conditions, porcine milk coagulated about 4 times faster than bovine milk and the resultant curd was about twice as soft. Dialysis of porcine milk against a large excess of bulk bovine milk did not influence its rennet coagulability nor did removal of 50% of its CCP, but complete removal of the CCP rendered porcine milk non-coagulable by rennin. Porcine milk was readily coagulated by rennin down to 5°C; its second-stage coagulation was independent of temperature – a characteristic due mainly to its high level of CCP.

Alcohol stability: porcine milk was coagulated by 66–68% (v/v) ethanol, as compared with 74% (v/v) for bovine milk.

Calcium stability: porcine milk was stable to at least 300 mM-CaCl2 at both 2 and 37°C.

Solvation: porcine milk contained significantly less water of hydration than bovine milk, 1·7g H2O/g casein compared with 2·0g H2O/g casein.

It was found that the increase in heat stability (coagulation time) that occurs when some milks are stored in light was not caused by changes in pH. These labile milks invariably gave a type A heat-stability curve, had a pH in the range corresponding to the coagulation-time minimum of the curve, and showed the expected abnormal 2-stage type of coagulation. The increase in coagulation time following exposure to light resulted from a delay in the occurrence of the first coagulation, the second coagulation time remaining unaltered. The presence of riboflavin in milk and of O2 in the headspace atmosphere during storage were necessary conditions for milk to be labile. Since light, headspace O2 and riboflavin had similar influences on rennet coagulation time it is suggested that in both instances the causative reaction, as proposed by others to account for the increase in rennet coagulation time of H2O2-treated milk, is stabilization of the heat- and rennin-sensitive methionine-phenylalaninebond in κ-casein by photo-oxidation of the methionine to methionine sulphoxide.

An examination was made of the effect of varying the protein, lactose and total serum ion content of milk on the rate of production of heat-induced acidity (as measured by pH decrease), on heat stability (as measured by coagulation time), on the relationship between coagulation time and initial pH of milk (as measured by coagulation time-pH curves) and on the interrelationship of these parameters. In addition, the effect on these parameters and their interrelationship of varying the composition and volume of the headspace atmosphere in contact with the milk was investigated. Explanations are proposed for the observed effects on heat stability of varying the composition of milk and the heating conditions, with special reference to the influence of heat-induced acidity.

The formation of protein polymers in ultra-high-temperature (UHT)-treated milk during storage at various temperatures was examined by gel filtration. The extent of polymer formation was found to depend on both storage time and temperature. After some months of storage at the higher temperatures of 30 and 37°C, the extent of polymerization of the caseins and whey proteins due to reactions of the Maillard type was several times greater than the heat-induced changes resulting from the UHT processing itself. After storage for 6 months the following proportions of milk proteins were found to exist in the form of covalently bound polymers: 50% at 37°C, 40% at 30°C, 26% at 20°C and 21% at 4°C. In addition, further amounts of polymer were formed by disulphide bonding, the contribution of such polymers diminishing gradually in a temperature-dependent manner during storage due to continuing polymerization reactions. It appeared that αs1-casein may be preferentially involved in these polymerizations with β-casein reacting at a somewhat slower rate. Polymerization and associated reactions modifying molecular charge led to the expected alterations in electrophoretic mobility and a loss of definition in the bands due to the various protein components. Measurements of proteolytic breakdown indicated only very small increases in trichloroacetic acidsoluble fragments, suggesting that proteolysis was of minor significance.

A comparison has been made of a number of different elution conditions for the separation by ion-exchange chromatography of the glycopeptides released by the action of rennin on whole milk. It was found that the most satisfactory separation was achieved using an NaCl gradient (0·10–0·18m) in 0·049 m-KH2PO4+0·001 m-Na2HPO4 at a constant pH of 5·1.

Data are presented which show that the use of glucono-δ-lactone as a chemical acidulant in cheese manufacture results in a much more rapid decrease in pH than that occurring in biologically acidified cheese. One of the consequences of the excessive early acid development was a very marked increase in the level of proteolysis during cheese-manufacturing operations. Consequently, chemically acidified cheese, as manufactured by the technique of Mabbitt, Chapman & Berridge (1955) (expt 17), does not appear to be a suitable ‘reference cheese’, especially for studies in which proteolysis during ripening is under investigation. The technique of Mabbitt et al. Avas modified to simulate the pH development pattern of starter cheese more closely; the level of proteolysis in curd made by the modified method was approximately equal to that in starter curd.

An attempt was made to link the propensity to develop bitterness of cheese made with fast acid-producing cultures with excessive early proteolysis in such cheese. The results show that fast- and slow-culture strains differed significantly in their acid-producing capability only after the cooking stage and consequently such cheeses differed little with respect to early proteolysis. Further, cheeses made with 0·5, 1·0, 2·0 or 4·0% of ML8 culture which differed considerably in rate of acid development and consequently in the level of early proteolysis did not develop bitterness, even when 4·0% starter was used.

It is concluded that gross proteolysis and bitterness are not correlated.

Yeast extract was fractionated on Sephadex G-25 into 7 fractions. The fraction most stimulatory to the growth of Streptococcus lactis C10 contained over 70% of the amino N present in yeast extract and consisted of a wide variety of free amino acids and a small amount of peptide material. Examination of possible replacement factors for this fraction revealed that the amino-acid material present was largely responsible for the stimulation of Str. lactis C10. Purine and pyrimidine bases and inorganic constituents also contributed to the stimulation. In addition, yeast extract contained a component which decomposed H2O2, a metabolite which accumulates in the growth medium under aerobic conditions and inhibits growth. The nature of the stimulation was studied by isolating slow and fast acid-producing colonies of Str. lactis C10. It appeared that yeast extract and other amino-acid supplements prevented an observed inhibition of the growth of the slow variants below pH 6·0, apparently by satisfying a nutritional deficiency caused by a drop in pH.

Citrate utilization and diacetyl, acetoin and acetaldehyde production by 2 strains each of Leuconostoc cremoris and Streptococcus diacetilactis in milk were studied. With the leuconostoc bacteria no growth and little citrate utilization occurred unless a stimulant (yeast extract) was present, when complete utilization of citrate without concomitant production of diacetyl or acetoin was obtained. The addition of Mn2+ stimulated growth and citrate utilization in the presence of yeast extract. Addition of citric acid after 65-h growth resulted in diacetyl and acetoin production. Destruction of diacetyl and acetoin occurred when the citric acid level fell to c. 1000 and 600 μg/g in the case of Leuc. cremoris FR8-1 and CAF1, respectively. Only strain FR8-1 produced acetaldehyde. In contrast, Str. diacetilactis produced diacetyl, acetoin and acetaldehyde concomitant with citrate utilization.

Several peptidase activities in the 3 species of Group N streptococci were partly separated by gel filtration on Sephadex G-200. The peptidases identified were a general aminopeptidase of wide specificity, a tripeptidase, a proline iminopeptidase (prolyl-β-naphthylamidase), a proline iminodipeptidase and an aminopeptidase-P. The effects of temperature and pH on the stability of the enzyme activities, and the influence of the type of N source used in the growth medium on the elution pattern of the enzymes were examined.

Buffalo αs-casein (α-sb) was found to be heterogeneous; on starch gelurea electrophoresis 4 electrophoretic components were detected.

The αsb-fraction was isolated by urea and alcohol fractionation. A preparation containing only the 2 main components was obtained by chromatography on DEAE-cellulose. Its amino acid pattern was different from that of cow αs1-casein. The fraction was also free of carbohydrate, cystine and cysteine.

Buffalo αsb-casein was less stable than cow αs-casein in solutions containing Ca, and the maximum stability of the αs-/κ-casein complex occurred at a lower Ca concentration.

β-Casein from individual buffalo's milk was found to be homogeneous by starch-gel electrophoresis. β-Casein was separated from buffalo's milk by the method of Warner (1944) and purified by DEAE-cellulose chromatography.

Buffalo β-casein possesses identical end-groups to those of cow β-casein; namely N-terminal arginine and assuming a single polypeptide chain a possible C-terminal sequence of Ile-Ile-Val. However, the amino-acid composition and the tryptic peptide patterns of the 2 proteins are not the same.

Although casein micelles are disrupted by removal of Ca, individual caseins remain aggregated in sub-micellar casein aggregates or sub-units. These sub-units have been studied by: (1) the use of gel filtration on Sepharose 4B at 6, 20 and 37°C at pH 6·7 and 0·1 ionic strength, (2) ultracentrifugation and (3) electron microscopy. At 37°C the protein composition of the sub-units varied across the gel-filtration peak, with κ-casein being eluted towards the leading edge and the ratio of αs1- to β-casein being almost constant across the peak. Re-chromatography of the protein from the leading edge of this peak gave a new wide peak with the κ-casein again being eluted towards the leading edge. However, αs1-casein was eluted before β-casein in the leading edge of the new peak. Prior treatment of the casein micelles by dispersion with 6 m-urea solution, precipitation with acid or reduction with 2-mercaptoethanol did not alter the gel-filtration pattern. An examination of the purified casein components and their mixtures showed that a 1:1 ratio mixture of αs1- and β-casein had the same peak maximum elution volume as casein micelle sub-units. κ-Casein by itself eluted at the void volume of the gel-filtration column, but after admixture with a sample of small micelles it eluted at the leading edge of the sub-unit peak and was indistinguishable from the κ-casein normally present. These results suggest that the sub-units are in equilibrium with their component caseins and that their size distribution is determined by only those factors (such as protein concentration, pH, temperature and ionic strength) which determine the strength of association between the casein components. The results from electron microscopy and ultracentrifugation support these conclusions.

Autoxidation was studied in milk obtained from cows fed on formaldehyde-treated casein-safflower oil supplement. The fat of this milk contained high levels of linoleic acid. A rapid disappearance of dissolved oxygen (DO), measured by using an oxygen electrode, from milk samples stored at 0°C in tubes without headspace, coincided with the development of oxidized flavours detected organoleptically. A correlation coefficient of 0·9 with 69 degrees of freedom (P < 0·001) was obtained between the amount of DO disappearing and the taste panel scores for oxidized flavours.

Butylated hydroxyanisole, sesamol, nordihydroguaiaretic acid, ethoxyquin, or butylated hydroxyanisole with propylgallate or tocopherols, when added in emulsified form to the milk at the rate of 10–15 mg/l milk, checked the development of oxidized flavours and the rapid disappearance of DO. Other antioxidants tested were either ineffective or imparted off-flavours to milk.

Samples of mare's milk neither developed oxidized flavours nor showed rapid disappearance of DO over a test period of 8 d.

The oxygen electrode provides a convenient and sensitive method for studying autoxidation and evaluating antioxidants in milk.