Lactobacillus bulgaricus strains LT1 and LT4 previously isolated from a yogurt factory were shown to be lysogenic and inducible by mitomycin C or u.v. irradiation, induction being optimal at the beginning of exponential growth. Five indicator strains (four of L. lactis, one of L. bulgaricus) were found in which the two phage lysates propagated well in a liquid medium, but formed plaques on bacterial lawns less readily. Ca2+ was required for the phage infection of the indicator strains to reach termination. Phage particles from each strain exhibited a comparable ultrastructural morphology under the electron microscope, having an isometric head, a triple collar and a noncontractile fibre-containing tail.

The aim of this Research Reflection is to describe the basic rumen function of goats and its modification in response to environmental factors, as well as to discuss similarities and differences when compared to other ruminants. In so doing we shall reveal the adaptive capacity of goats to harsh environments. The basic rumen function in goats is similar to other species of ruminants, as stressed by the opportunity to apply the updates of feeding systems for ruminants to goats. The rumen epithelium acts as a protective barrier between the rumen and the host, but it can be damaged by toxic compounds or acidosis. The rumen also plays an important role in water balance, both for dehydration and rehydration. Recent studies show that the microbiota exhibits a high fractional stability due to functional redundancy and resilience, but this needs more investigation. The microbial community structure differs between goats and cows, which explains the difference in sensitivity to milk fat depression following intake of high lipid diets. Goats also differ from other ruminants by their enhanced ability to feed-sort, but as with cows they can suffer from acidosis. Nevertheless, goats can be considered to be very resistant to environmental factors such as water stress, salt stress or heat stress, and this is especially so in some endogenous breeds. They also are able to detoxify tannins, polyphenols and other secondary metabolites. Some new trials involving feeding behaviour, microbiota and omics or approaches by meta-analyses or modelling will improve our knowledge of rumen function in goats.

Ovine casein macropeptide (CMP) was characterized by anion-exchange FPLC and reversed-phase (RP) HPLC. To study heterogeneity (the degree of glycosylation and phosphorylation), CMP was desialylated with neuraminidase and dephosphorylated with acid phosphatase. Following RP-HPLC, the main CMP components were identified using either on-line or off-line mass spectrometry. The most abundant ovine CMP component was a diphosphorylated carbohydrate-free form, followed by one or two monophosphorylated and a non-phosphorylated asialo-aglyco species. Aglyco non-phosphorylated, monophosphorylated and diphosphorylated forms were in the ratio 3[ratio ]20[ratio ]77. Only ∼ 30% of ovine CMP was glycosylated. Assuming that the monosaccharide fraction of ovine CMP is composed of N-acetylgalactosamine, galactose and N-glycolylneuraminic acid, molecular masses consistent with the presence of CMP containing tetra-, tri-, di- and monosaccharide were identified.

Dairy cows fitted with ruminal, duodenal and ileal cannulas were utilized to investigate the effects of feeding with Ca soaps (CaS) of palm fatty acids (FA) and rapeseed FA. Diets compared were control diet based on maize silage and concentrate, and two diets with 40 g CaS of palm oil FA or rapeseed oil FA/kg diet, replacing part of the concentrates of the control diet. Total digestibilities of dry matter, fibre and fat, and ruminal fermentation were not significantly altered by giving CaS; the extent of ruminal biohydrogenation of total unsaturated C18 FA was significantly reduced by both CaS diets. Apparent intestinal digestibility of FA was not different among diets, although the amount of FA absorbed with the CaS diets was twice that with the control diet. No difference among diets was observed for milk production, or fat and protein contents. Giving CaS diets decreased the proportions of 4[ratio ]0 to 14[ratio ]0 FA in milk fat, and increased cis-18[ratio ]1n−9, compared with control diet. The rapeseed diet lowered the content of 16[ratio ]0, and increased the contents of 18[ratio ]0 and trans-18[ratio ]1n−7. CaS diets did not result in a marked increase of polyunsaturated FA content in milk fat. Butter from cows fed on the CaS diets contained more liquid fat at 6 and 14°C than butter from the cows fed on the control diet. Incorporating CaS, particularly those from rapeseed, in dairy cows' diets increased C18 FA in milk and improved butter spreadability.

Bacterial analyses were carried out of 2069 udder secretions, isolated from 1481 heifers with mastitis in eight veterinary districts in Sweden. Streptococci, e.g. Streptococcus dysgalactiae and Str. uberis, dominated the bacterial flora, being isolated from 34·4 and 19·5% respectively of heifers with clinical mastitis occurring from puberty up to 14 d post partum. Bacterial species generally regarded as important pathogens in the summer mastitis complex, e.g. Actinomyces pyogenes, Stuart–Schwan coccus and strictly anaerobic bacteria such as Peptostreptococcus indolicus, Fusobacterium necrophorum and Bacteroides melaninogenicus were isolated at low frequencies (13·2, 6·3, 9·4, 3·8 and 1·3% respectively). When the cases of mastitis studied were restricted to those appearing in heifers pre partum, May 15 to October 14 (summer mastitis), these bacterial species were isolated at higher percentages (27·1, 14·4, 21·4, 13·5 and 5·2% respectively). These figures were, nevertheless, still lower than those published in reports from other countries. Whether considered up to 14 d post partum or only pre partum, there were no significant differences in the frequencies of A. pyogenes isolated at different seasons. There were geographical differences in bacterial incidence, e.g. Staphylococcus aureus was isolated significantly more often in northern regions whereas Str. dysgalactiae was more common in the south. This and other Swedish investigations support the theory that A. pyogenes and strictly anaerobic bacteria are ‘secondary invaders’ that depend on Str. dysgalactiae to cause a primary infection. It is stressed that the udders of all heifers should be examined daily so that cases of mastitis can be treated immediately.

This study describes a method for species-specific detection of animal DNA from different species (cattle, sheep, goat, water buffalo) in milk and dairy products. A primer set was designed in conserved region on the basis of the alignment of the sequence codifying the genomic κ-casein gene in order to amplify all four species with a single primer pair. Polymorphisms were detected via minisequencing with extension primers designed in conserved sequences for haplotype determination that allow unambiguous assignment to each species. The method was successfully applied to the detection of raw and pasteurized milk from the four different species considered as well as to cheese products from the retail trade. Estimation of the limit of detection was carried out using a progression of dilutions of genomic DNA as well as DNA isolated from milk of a known number of somatic cells from different species in order to be able to achieve detection rates as low as 0·1% bovine milk mixed with buffalo milk.

Stereospecific analysis is an important tool for the characterization of lipid fraction of food matrices, and also of milk samples. The results of a chemical-enzymatic-chromatographic analytical method were elaborated by chemometric procedures such as linear discriminant analysis (LDA) and artificial neural network (ANN). According to the total composition and intrapositional fatty acid distribution in the triacylglycerol (TAG) backbone, the obtained results were able to characterize pure milk samples and milk mixtures with 1, 3, 5% cow milk added to donkey milk. The resulting score was very satisfactory. Totally correct classified samples were obtained when the TAG stereospecific results of all the considered milk mixtures (donkey-cow) were elaborated by LDA and ANN chemometric procedures.

The objective was to determine the proportions and bacterial counts of major mastitis pathogens in samples of bulk tank milk (BTM), as well as to clarify the relationship between these bacteria counts and bulk tank somatic cell count (BTSCC). The purpose was to judge the importance of the counts of mammary pathogens for BTSCC at the population level. Samples of BTM were collected from 268 randomly selected anonymous dairy herds (with approximately 29 000 dairy cows). Staphylococcus aureus, other coagulase-positive staphylococci, Streptococcus agalactiae and Streptococcus dysgalactiae were grouped as contagious pathogens, and Streptococcus uberis, Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis and Enterococcus faecium as environmental pathogens. Based on assessment of the dominant pathogen in each herd, environmental pathogens were found to dominate. Counts of specific mammary pathogens in BTM samples did not exceed a geometric mean of 1000 cfu/ml. Significant differences were documented in BTSCC between BTM samples containing Str. agalactiae, Staph. aureus, and Str. dysgalactiae and BTM samples that were pathogen-free. Geometric means of BTSCC associated with these pathogens were noticeably higher than the overall BTSCC (Str. agalactiae 243 628; Staph. aureus 205 610; Str. dysgalactiae 203 978; overall 173 000 cells/ml). It follows that the somatic cell count (SCC) associated with these pathogens contributed substantially to the overall BTSCC. Environmental pathogens predominated in samples of BTM with SCC <300×103/ml and contagious mastitis pathogens predominated in BTM samples with SCC >300×103/ml. No correlation was detected between bacterial counts of specific pathogens and BTSCC. This study revealed that the assessment of bacterial counts of mammary pathogens in samples of BTM in relation to BTSCC is applicable for the monitoring of changes in the occurrence of major mastitis pathogens in dairy herds at a national level.

Some of the acid phosphatase isozymes of bovine leucocytes and plasma have been separated and partly characterized. About 80% of the phosphatase activity of leucocytes at pH 4·9 was particle-bound and about 8% was extractable with Amberlite CG-50 ion exchange resin. This extractable enzyme existed as a single electrophoretic component with a mol. wt of about 42000 and with optimum activity at pH 5·8. Km for p-nitrophenyl phosphate was 1·6 mM at pH 5·8 and 0·4 mM at pH 4·9. At pH 5·8 orthophosphate (K1 = 1·5 mM) and pyrophosphate (Ki = 4·1 mM) were competitive inhibitors. The enzyme was also strongly inhibited by F−, Al3+, IO4− and S2032−. The enzyme which was not extractable with Amberlite was very heterogeneous with respect to molecular weight. At the pH optimum (4·9), Km for p-nitrophenyl phosphate was 0·4 mM and orthophosphate (K1 = 2·3 mM) and pyrophosphate (K1 = 2·1 mM) were competitive inhibitors. Other inhibitors included F−, Al3+, Hg2+, IO4− and tartrate. The enzyme extracted from plasma by Amberlite CG-50 treatment had properties similar to that extracted from leucocytes. Normal bovine milk contained a single acid phosphatase, but milk from cows with mastitis showed 3 electrophoretic isozyme bands, one being the same as in normal milk; the 2 additional bands were of leucocyte origin.

1. The variation in the stability of milk protein to heat and the relationship between milk composition and heat stability were examined.

2. The coagulation times of the majority of the milk samples decreased by a factor of about 3 with an increase in temperature of 10°C. over the range 130–150°C. Because of the general proportionality of the coagulation times at 130, 140 and 150°C., the coagulation time at 130°C. only were used as a measure of the stability of the samples to heat.

3. The coagulation times of herd bulk milks ranged from 17·2 to 59·0 min. at 130°C., whereas the range for samples from individual cows was 0·6–86·2 min.

4. Samples of colostrum were very unstable to heat, and milk from cows in late lactation tended to have the longest coagulation times, but otherwise there was little relation between the heat stability of milk and the stage of lactation of the cow.

5. Although colostrum samples were comparatively rich in ionized calcium, their marked instability to heat appeared to be caused solely by their high content of lactalbumin plus lactoglobulin.

6. The stability to heat of the calcium caseinate-calcium phosphate complex in all samples, other than colostrum, could not be closely related either to the concentration and composition of the complex or to the composition and salt-balance of the aqueous phase.

7. When the calcium phosphate content of the caseinate complex was relatively low, the heat stability of the complex tended to be inversely related to the concentration of ionized calcium in the milk, but in general coagulation time was not related to the concentration of ionized calcium.

A simplified moving boundary electrophoresis technique has been developed for the measurement of the electrophoretic mobility of casein micelles. The zeta potentials of casein micelles from different skim-milk samples were calculated using Henry's equation and shown to decrease with decrease in pH between pH 6.9 and 5.3 and to increase with increase in temperature between 10 and 45 °C. Neither severe heat treatment (up to 135 °C for 51 min) nor centrifugal fractionation of micelles into different micelle size ranges had any significant effect on zeta potential. The ionic composition of the serum phase has been shown to be extremely important in determining the electrophoretic mobility. Casein micelles electrophoresed through milk ultrafiltrate consistently gave a lower mobilities than the same micelles centrifuged through milk centrifugate. The results are discussed in relation to present theories of casein micelle structure; these theories do not accommodate all of the observations.

Four grade Jersey cows were exposed for 7 hr. twice a week to each of several atmospheres having dry-bulb temperatures ranging from 85 to 110° F. and absolute humidities ranging from 6 to 16 gr./cu.ft. The following reactions were studied: rectal temperature; pulse rate; respiratory rate; respiratory volume; evaporative loss; calcium, phosphate and erythro-cyte composition of the blood; milk volume and butterfat content; behaviour.

Rectal temperature rose to higher values with less ready establishment of equilibrium the hotter the condition, but exceeded 107° F. only in the hottest atmosphere. (Dry-bulb temperature 110° F., absolute humidity 16 gr./cu.ft.)

Respiratory rate was similarly and markedly affected. In both cases, humidity had a marked effect as well as temperature, an increment of 0–4 gr./cu.ft. (approx. 4%) in humidity having the same effect as 1° F. rise in air temperature. The highest average respiratory rate was 200/min. Respiratory minute volume rose less than the rate, so that tidal volume was reduced.

Pulse rate was essentially unaffected by a rise in temperature but tended to rise somewhat with humidity.

Evaporative loss was markedly increased by temperature, much less so by humidity. Increased pulmonary ventilation was inadequate to account for the observed losses, which are attributed to sweat-gland activity.

Behavioural changes included some licking, panting, salivation, mild agitation, cessation of rumination and refusal of water. Weakness and staggers did not occur under these conditions.

Neither milk nor butterfat production was essentially affected by the exposures. Blood calcium and phosphate levels fell, but the erythrocyte count was unchanged.

These findings provide basic data with which subsequent observations under varying conditions can be compared.

1. The composition of the milk of six low-plane and six high-plane ewes has been studied. Definite lactational trends have been observed for all the major constituents.

2. For all the ewes the initial colostrum was characterized by very high protein and fat contents and by low lactose content.

3. Analyses of the mineral constituents—calcium, magnesium, sodium, potassium, phosphorus, and chloride—are reported covering the whole lactation of twelve sheep. The colostrum of the low-plane animals was higher in total mineral content than that of the high-plane animals.

The author wishes to thank Mr L. J. Lambourne for providing the milk samples and Mr J. E. Allen for the spectographic analyses.

T cells play a central role in specific immunity; their populations and phenotypes could be affected by number of lactation in high-yielding dairy cows. To investigate the effects of parity on the dynamics of T lymphocytes, lymphoproliferative capacity, T lymphocyte subsets and CD4+/CD8+ ratio were studied in peripheral blood of primiparous and pluriparous dairy cows during mid–late lactation. A non-radioactive technique was also adapted for a detailed lymphoproliferation assay. Compared with the primiparous cows, the pluriparous cows exhibited weaker lymphoproliferative activity, larger number of CD4+ cells and substantially greater CD4+/CD8+ ratio in their blood circulation. The increase of the CD4+/CD8+ ratio in the blood of pluriparous dairy cows was mainly due to the rise in the proportion of CD4+ cells and decline in the proportion of CD8+ cells. This increase of the CD4+/CD8+ ratio coincided with the decrease of mitogen-induced proliferation capacity of T lymphocytes. Of four lymphocyte divisions or generations during the lymphoproliferation assay, maximal lymphocyte proliferation capacity at generation 3 in primiparous cows was markedly greater than in pluriparous cows. With an alternatively safer, faster and more reproducible assay (compared with 3H-thymidine scintillation assay) we showed for the first time that aging in dairy cows leads to a decreased mitogen-induced lymphoproliferation and disturbed proportion between CD4+ and CD8+ T cells. This CD4+-CD8+ imbalance together with diminished lymphoproliferative capacity may lead to a weaker T cytotoxic-mediated immunity and increased susceptibility to infectious diseases in pluriparous lactating cows. Our study also emphasizes further application of the methods in farm animals.

The C6–C11 2,4–dienals have been identified in distillates from skim milk with oxidized flavour. The C4–C11 2–enals were isolated from the same distillates in a previous investigation. The most abundant individual compounds in the two groups were 2–octenal, 2–nonenal, 2–4–heptadienal and 2,4–nonadienal. These compounds when flavour–tested in skim milk in dilutions of 10–7–10–9 closely resembled oxidized (cardboard) flavour. It is concluded that they are the main constituents of this flavour defect and that they originate from the oxidation of the more highly unsaturated fatty acids in milk lipids.

Average N-acetyl neuraminic-acid (NANA) content of milk from cow, sheep, goat, mare, donkey and sow (about 10 samples from each species) was 17·7, 10·1, 6·4, 6·1, 12·0 and 64·0mg/100 ml, of which 14·2, 8·4, 3·2, 3·1, 5·0 and 53·7mg/100 ml respectively was in the casein fraction. The NANA content of casein showed marked species variation ranging from an average of 2·18 % for the sow to 0·09 % for the goat.

About 28 % of the casein NANA of mare's milk was liberated in the fraction soluble in 12 % TCA by the action of rennin at pH 6·6 compared with 75 and around 100 % for donkey's and sow's milk respectively; about half the total NANA in the casein of cow's, sheep's and goat's milk was released under the same conditions.

Dried skim-milk stored at 55°C in air at 70% relative humidity has been found to contain lactulose, galactose, tagatose, glyceraldehyde and maltol; and formic, acetic and glycollic acids. These compounds are not present, or are present only in trace quantities, in fresh dried skim-milk. It is suggested that while the proteinsugar reaction is responsible for most of the browning of the powder, many of the compounds found are formed by degradation of lactose catalysed by the free basic amino groups of the casein. The compounds so formed probably then react with amino groups in a Maillard reaction and thus contribute to the browning of the milk powder.

The development of a rapid, highly sensitive double antibody–solid phase (DASP) radioimmunoassay for ovine/caprine prolactin is described. The assay was applied to the measurement of basal levels of prolactin in the blood of 4 anoestrous lactating female, 4 anoestrous virgin female and 4 castrated male goats. For several hours after milking there was a chronic gradually decreasing release of prolactin in the lactating goats, such that the circulating levels of prolactin were, for 14–16 h of the day, at a much higher level than those in the virgin female or castrated male goats. The basal levels of prolactin in the castrated male goats (87–200ng/ml) were somewhat higher than those in the virgin females (47–77ng/ml). The effects of feeding and stress on prolactin levels are discussed.

1. Microbiological methods were used to measure biotin, nicotinic acid, pantothenic acid, riboflavin, ‘vitamin B6’ and vitamin B12 in milk from Guernsey, Friesian and Shorthorn cows.

2. A study was made of day-to-day variations and breed differences in the B-vitamin content of such milk. Fluctuations in each vitamin were observed from one day to the next in milk from individual cows, these variations being especially marked with biotin and vitamin B12. In addition, the variations in the mean vitamin levels of milk from cows of the same breed were generally greater than any breed difference.

3. Samples of milk from ten Friesian and ten Shorthorn cows were taken at intervals throughout one complete lactation and analysed for each of the vitamins mentioned above. The results of this study of lactational trends are presented in detail.