The variable content of human breast milk suggests that its routine fortification may result in sub-optimal nutritional intakes and growth. In a pragmatic trial, we randomised infants born below 30 weeks of gestation to either the intervention (Igp) of fortifying milk on measured composition according to birth weight criteria and postmenstrual age (PMA) or our routine practice (RPgp) of fortifying on assumed milk composition to target 3·8–4·4 g protein/kg per d and 545–629 kJ/kg per d. Milk composition was measured using the MIRIS® Human Milk Analyser. Percentage fat mass (%FM) was measured using PEA POD (COSMED). The effects of macronutrient intakes and clinical variables on growth were assessed using mixed model analysis. Mean measured protein content (1·6 g/100 ml) was higher than the assumed value (1·4 g/100 ml), often leading to lower amounts of fortifier added to the milk of intervention infants. At discharge (Igp v. RPgp), total protein (3·2 (sd 0·3) v. 3·4 (sd 0·4) g; P=0·067) and energy (456 (sd 39) v. 481 (sd 48) kJ; P=0·079) intakes from all nutrition sources, weight gain velocity (11·4 (sd 1·4) v. 12·1 (sd 1·6) g/kg per d; P=0·135) and %FM (13·7 (sd 3·6) v.13·6 (sd 3·5) %; P=0·984) did not significantly differ between groups. A protein intake >3·4 g/kg per d reduced %FM by 2 %. Nutrition and growth was not improved by targeting milk fortification according to birth weight criteria and PMA using measured milk composition, compared with routine practice. Targeting fortification on measured composition is labour intensive, requiring frequent milk sampling and precision measuring equipment, perhaps reasons for its limited practice. Guidance around safe upper levels of milk fortification is needed.

Human milk proteins provide term and preterm infants with both nutrition and protection. The objective of the present study was to examine longitudinal changes in the protein composition of term and preterm milk during the first 2 months of lactation, focusing on protein phosphorylation and glycosylation. Using gel electrophoresis, the relative concentration and glycosylation status of lactoferrin, secretory Ig A, β-casein, α-lactalbumin, serum albumin, bile salt-stimulated lipase, xanthine oxidoreductase, tenascin and macrophage mannose receptor 1 were measured in milk collected on days 7, 10, 14, 18, 21, 28 and 60 postpartum from preterm mothers (28–32 weeks gestation, n 17). The phosphorylation status of β-casein was also investigated. To determine if these variables differ in term and preterm milk, samples from term mothers (38–41 weeks gestation, n 8) collected on days 7, 14 and 30 of lactation were also analysed. The concentration of the abundant milk proteins decreased during lactation in term and preterm milk (P <0·05). No difference in protein glycosylation was observed, except for the glycoproteins serum albumin and tenascin. The phosphorylation of β-casein varied significantly between term and preterm milk. Further investigation is required to determine whether these modifications affect protein function and are clinically important to preterm infants.

Donor human milk is pasteurized to prevent the potential risk of the transmission of pathogens to preterm infants. Currently, Holder pasteurization (human milk held at 62·5°C for 30 min) is used in most human milk banks, but has the disadvantage that it results in excessive inactivation of important bioactive components. Power-ultrasound (20–100 kHz) is an emerging technology for the preservation of foods and could be an alternative method for the treatment of human milk. The aim of this study was to investigate the effect of different ultrasound settings on the elimination of Escherichia coli and the retention of bile salt stimulated lipase (BSSL) activity. Ultrasonication with a constant power decreased Esch. coli viability exponentially over time until the processing temperature increased to sub-pasteurization level to between 51·4 and 58·5°C, then a log10 1·3 decrease was observed (P<0·05). BSSL activity decreased to 91% until a temperature of 51·4°C and then it decreased to 8% between 51·4 and 64·9°C. Ultrasonication with a constant energy and various power and exposure times showed the highest temperature (53·7°C) when treated with the longest exposure time and lowest ultrasound-power (276 s at 3·62 W) compared with 37·6°C for 39 s at 25·64 W. The findings predict that the viability of Esch. coli could be reduced by log10 5 with a minimal loss of activity of BSSL by applying 13·8 kJ of energy in 12 ml of human milk using high ultrasound power over a short exposure time to ensure that the temperature remains below the critical level for protein denaturation. Alternatively, the use of lower power settings such as the 26 W used in the present studies would require a cooling system to ensure the human milk BSSL was protected against temperature denaturation.

The combination of ultrasound and heat (thermoultrasound) is an emerging food preservation technique that retains higher quantities of bioactive components compared with current thermal pasteurization practice, but has not yet been assessed for pasteurizing human milk. Artificially contaminated human milk samples were treated with ultrasound (20 kHz, 150 watts) with and without heating. The retention of four human milk proteins was quantified by biochemical assay and laser scattering particle sizing was used to determine the extent of homogenization. While ultrasonic treatment was effective at inactivating Escherichia coli (D4 °C=5·94 min), Staphylococcus epidermidis exhibited resistance (D4 °C=16·01 min). Thermoultrasonic treatment was considerably more effective (Esch. coli D45 °C=1·74 min, D50 °C=0·89 min; Staph. epidermidis D45 °C=2·08 min, D50 °C=0·94 minutes) with a predicted retention (2·8 min treatment, 50°C) of secretory IgA lysozyme, lactoferrin and bile salt stimulated lipase of 91, 80, 77, and 45%, respectively. Homogenization of the milk samples occurred after 5 min and 2 min of ultrasonic and thermoultrasonic treatment, respectively. Thermoultrasonic treatment is an effective method for pasteurizing donor human milk and retaining a greater proportion of bioactive components compared with current practices. However, further studies are required to assess the practicality of applying this technique routinely to donor human milk.

The evolutionary origin of the mammary gland has been difficult to establish because little knowledge can be gained on the origin of soft tissue organs from fossil evidence. One approach to resolve the origin of lactation has compared the anatomy of existing primitive mammals to skin glands, whilst another has examined the metabolic and molecular synergy between mammary gland development and the innate immune system. We have reviewed the physiology of lactation in five mammalian species with special reference to these theories. In all species, milk fulfils dual functions of providing protection and nutrition to the young and, furthermore, within species the quality and quantity of milk are highly conserved despite maternal malnutrition or illness. There are vast differences in birth weight, milk production, feeding frequency, macronutrient concentration, growth rate and length of lactation between rabbits, quokkas (Setonix brachyurus), pigs, cattle and humans. The components that protect the neonate against infection do so without causing inflammation. Many protective components are not unique to the mammary gland and are shared with the innate immune system. In contrast, many of the macronutrients in milk are unique to the mammary gland, have evolved from components of the innate immune system, and have either retained or developed multiple functions including the provision of nourishment and protection of the hatchling/neonate. Thus, there is a strong argument to suggest that the mammary gland evolved from the inflammatory response; however, the extensive protection that has developed in milk to actively avoid triggering inflammation seems to be a contradiction.

Despite the importance of human milk fatty acids for infant growth and development, there are few reports describing infant intakes of individual fatty acids. We have measured volume, fat content and fatty acid composition of milk from each breast at each feed over a 24h period to determine the mean daily amounts of each fatty acid delivered to the infant from breast milk at 1, 2, 4, 6, 9 and 12 months of lactation in five women. Daily (24h) milk production was 336·60 (sem 26·21) and 414·49 (sem 28·39) ml and milk fat content was 36·06 (sem 1·37) and 34·97 (sem 1·50) g/l for left and right breasts respectively over the course of the first year of lactation. Fatty acid composition varied over the course of the day (mean CV 14·3 (sd 7·7) %), but did not follow a circadian rhythm. The proportions (g/100g total fatty acids) of fatty acids differed significantly between mothers (P<0·05) and over the first year of lactation (P<0·05). However, amounts (g) of most fatty acids delivered to the infant over 24h did not differ during the first year of lactation and only the amounts of 18:3n-3, 22:5n-3 and 22:6n-3 delivered differed between mothers (P<0·05). Mean amounts of 18:2n-6, 18:3n-3, 20:4n-6 and 22:6n-3 delivered to the infant per 24h over the first year of lactation were 2·380 (sd 0·980), 0·194 (sd 0·074), 0·093 (sd 0·031) and 0·049 (sd 0·021) g respectively. These results suggest that variation in proportions of fatty acids may not translate to variation in the amount delivered and that milk production and fat content need to be considered.

We developed a simple, non-invasive method to measure the intramammary pressure (IMP) response in sows during natural milk ejection and after i.v. injections of oxytocin. The teats of sows were cannulated and the amplitude and duration of IMP changes were measured via a pressure transducer linked to a computer. During natural milk ejections, the amplitude of the response was 5·5±1·8 kPa (mean±SD) and the duration of response at half the peak amplitude was 6·8±1·6 s. In response to oxytocin a threshold response occurred at 60 mIU oxytocin with no further change up to 200 mIU. The amplitude of the response to an i.v. dose of 60–200 mIU oxytocin (5·5±2·2 kPa) and duration (6·4±1·7 s) were similar to that during a natural milk ejection. The responses of two galactophorous ducts of a teat measured simultaneously were similar in peak amplitude and duration. When the responses of four glands of a sow were measured simultaneously, there was a lag time in the onset of the IMP peak from the anterior to the posterior glands, and an increase in the peak amplitude from anterior to posterior. Both during natural milk ejections and in response to oxytocin injections, there were significant differences in peak amplitude between sows. We also aimed to determine whether the pressure or duration of milk flow during natural milk ejections was limiting piglet growth rates. IMP of nine sows and the growth rates of their piglets were measured. Both the peak amplitude of IMP response and the growth rate of piglets increased over the course of lactation, but the relationship was not causal. We found that the growth rate of piglets did not depend on IMP amplitude or duration, but we suggest that it may depend on the frequency of suckling and/or the ability of piglets to withdraw milk while it is available.

Fat in human milk is extremely variable and can represent up to 50 % of infant energy intake. To accurately determine milk composition and infant intake at 1 (n 17), 2 (n 17), 4 (n 17), 6 (n 15), 9 (n 6) and 12 (n 5) months of lactation, samples of fore- and hind-milk were collected from each breast at each feed over 24 h periods from an initial group of seventeen women. The content of fat in milk varied over 24 h, with a mean CV of 47·6 (SE 2·1) % (N 76) AND 46·7 (se 1·7) % (n 76) for left and right breasts respectively. The 24 h amounts of fat, lactose and protein in milk differed between women (P=0·0001), but were consistent between left and right breasts. Daily milk production differed between breasts (P=0·0001) and women (P=0·0001). Accordingly, amounts of fat (P=0·0008), lactose (P=0·0385) and protein (P=0·0173) delivered to the infant over 24 h also differed between breasts and women (P=0·0001). The energy content of milk and the amount of energy delivered to the infant over 24 h were the same between breasts, but differed between women (P=0·0001). The growth rate of a group of only six infants in the present study was not related to either the concentrations or amounts of fat, lactose, protein and energy in milk over the first 6 months of life. These results show the individuality of milk composition and suggest that only a rigorous sampling routine that takes into account all levels of variation will allow the accurate determination of infant intake of fat, lactose, protein and energy.

Previous studies have suggested that the uptake of prolactin from the blood into the milk may be restricted when the alveolus is distended with milk. Therefore the aim of this study was to determine the relationship between prolactin in milk and milk production by measuring the concentration of prolactin in samples of fore- and hind-milk as well as the volume of milk removed for each breast, at each breastfeed over a 24 h period. The mean (± S.D.) concentration of prolactin in milk for all women (n = 15) over the 24 h period was 18.5 ± 11.6 µg l-1 (fore-milk) and 16.8 ± 12.8 µg l-1 (hind-milk). The variation between women masked small changes within women in the concentration of prolactin in milk over the 24 h period, therefore a prolactin ratio (individual fore- or hind-milk value divided by the mean for all fore- or hind-milk samples collected over a 24 h period) was determined. The concentration of prolactin was highest in milk between 02.01 and 06.00 h (prolactin ratio for fore- to hind-milk, 1.5), and lowest between 10.01 and 18.00 h (prolactin ratio for fore- to hind-milk, 0.8). Furthermore, we observed that the difference in prolactin concentration between the fore- and hind-milk (fore-hind gradient) was greatest between 06.01 and 10.00 h (4 µg l-1). To ensure that this effect was not due to permeability in the paracellular pathway, the concentrations of serum albumin and sodium in milk were measured. No significant (P > 0.05) changes over the 24 h period, or with increasing time since last feed were observed. We therefore concluded that when the breast is most drained of milk (in the late evening), and the rate of milk synthesis is greatest, the fore-hind prolactin gradient in the milk of the following feed (in the early morning) is highest. This is consistent with the observation that prolactin uptake from the blood by the lactocyte is dependent on the fullness of the breast, such that prolactin uptake may be inhibited in full alveoli. Experimental Physiology (2002) 87.2, 207-214.

Breast volume and morphology of eight subjects were measured before conception and at intervals throughout pregnancy until 1 month of lactation. Breast volume before conception ranged from 293 to 964 ml. At the end of pregnancy the volume of breast tissue had increased by 145 ± 19 ml (mean ± S.E.M., n = 13 breasts, range 12-227 ml) with a further increase to 211 ± 16 ml (n = 12 breasts, range 129-320 ml) by 1 month of lactation. Urinary excretion of lactose increased at 22 weeks of pregnancy, signalling the capacity of the breast to synthesize lactose at this time. During pregnancy, both the change in breast volume and the change in cross-sectional area of the areola were related to the concentration of human placental lactogen in the plasma. The growth of the nipple and the rate of excretion of lactose were related to the concentration of prolactin in the plasma. During the first 3 days after birth, the rate of excretion of lactose was related to the rate of excretion of progesterone. There was no relationship between the growth of the breast during pregnancy and the amount of milk produced at 1 month of lactation.

Quantitative measurements were made of relative breast volume and milk production from 1 month of lactation until 3 months after weaning, and the storage capacity of the breasts was calculated. The increase in breast tissue volume from before conception until 1 month of lactation was maintained for the first 6 months of lactation (means ± S.E.M.) (190.3 ± 13.1 ml, number of breasts, nb = 46). During this period of exclusive breast-feeding, 24 h milk production from each breast remained relatively constant (453.6 ± 20.1 g, nb = 48), and storage capacity was 209.9 ± 11.0 ml (nb = 46). After 6 months, breast volume, milk production and storage capacity all decreased. There was a relationship between 24 h milk production and the storage capacity of the breasts, and these both appeared to be responding to infant demand for milk. At 15 months of lactation, the 24 h milk production of each breast was substantial (208.0 ± 56.7 g, nb = 6), even though the breasts had returned to preconception size. This was associated with an apparent increased efficiency of the breast (milk production per unit breast tissue) after 6 months, which may have been due to redistribution of tissues within the breast. The possible causes of the decrease in breast volume are discussed.

Colostrum and milk were collected from ten sows at frequent intervals from before farrowing until 9 d after farrowing. Ionized calcium, pH, and total concentrations of citrate, calcium, phosphate and magnesium were measured in whole milk. The diffusible fraction of the mammary secretion was separated by ultrafiltration and was used for the measurement of diffusible citrate, calcium, phosphate and magnesium. The pH before farrowing was 5·7, and increased to 6·5 on day 4 as total calcium and phosphate also increased. Before farrowing, total and diffusible citrate were 7·8 and 7·3 mM respectively, while diffusible phosphate was 11·9 mM, and these concentrations all decreased during the study period. Total magnesium ranged between 3·3 and 4·1 mM, while diffusible magnesium ranged between 2·0 and 3·1 mM. While these concentrations and patterns of change of diffusible calcium and citrate are quite different from those of women's milk during the first week after birth, theoretical physicochemical relationships between diffusible calcium and citrate, and ionized calcium and HPO42− were corroborated by these results. We conclude that diffusible citrate plays an important role in the determination of the concentration of diffusible calcium. However, while citrate may be the major determinant of the total concentration of calcium in women's milk, this is not the case in sows' milk.

The aims of the present study were (a) to maintain the structure and function of the small intestine of the piglet after weaning, and (b) to compare the capacity in vivo of sucking and weaned piglets to digest oral boluses of lactose and sucrose and absorb their monosaccharide products. Piglets were fed on cows' whole milk ad libitum every 2 h for 5 d after weaning. Physiological doses of lactose plus fructose (treatment LAC + FRU) and sucrose plus galactose (treatment SUC + GAL) were administered on day 27 of lactation and on the fifth day after weaning, after which time piglets were killed. Villus height and crypt depth were maintained (P > 0·05) by feeding cows' milk after weaning. The areas under the curves (AUC) for galactose and glucose, adjusted for live weight and plasma volume, increased (P < 0·05) after weaning. Despite the enhancement of gut function after weaning, the galactose index (Gall: AUC for galactose ingested as lactose divided by the AUC for the same dose of galactose ingested as the monosaccharide) and fructose index (FruI: AUC for fructose ingested as sucrose divide by the AUC for the same dose of fructose ingested as the monosaccharide), which are indices of digestive and absorptive efficiency, both decreased after weaning. This apparent anomaly may be reconciled by increased growth, and hence surface area, of the small intestine between weaning and slaughter such that ‘total’ digestion and absorption most probably increased despite apparent decreases in GalI and FrnI. Positive correlations (P < 0.05) between villus height and Gall are consistent with the maximum activity of lactase occurring more apically along the villus. Significant linear relationships (P < 0·05) were recorded between villus height at the proximal jejunum and adjusted AUC for galactose and glucose following treatment LAC + FRU, and between villus height at the proximal jejunum and adjusted glucose AUC following treatment SUC + GAL. These relationships suggest that maximum digestion and absorption occurs at increasing distances along the crypt:villus axis in the weaned pig.

The concentrations of lactose, glucose, glucose 6-phosphate, glucose 1-phosphate, UDPglucose, UDPgalactose, UDP, UMP, inorganic phosphate, ADP and AMP (metabolites involved in the lactose synthesis pathway), and cAMP, galactose and fructose were measured in the mammary secretion from sucked (n = 9) and unsucked (n = 4) mammary glands of nine sows during the first 5 d post partum. The concentrations of lactose, glucose, galactose and fructose were also measured in plasma during this time. The progressive increase in the concentration of lactose, and changes in the concentrations of cellular metabolites in the mammary secretion from sucked glands were consistent with an increase in the metabolic activity of those glands during lactogenesis II. In contrast, unsucked glands showed a progressive decrease in the concentration of lactose, while the concentrations of cellular metabolites in the milk generally remained unchanged. These results indicated that there was no increase in the metabolic activity of unsucked glands (no increase in lactose synthesis or utilization of glucose and ATP) and that the rate of lactose synthesis prior to milk removal was limited by the availability of glucose and/or UDPgalactose. Therefore, the removal of colostrum from the mammary gland was necessary for an increase in the rate of lactose synthesis (and probably de novo fatty acid synthesis) and implies that autocrine mechanisms are operating to control the rate of milk synthesis during lactogenesis in the sow. The low concentration of glucose in colostrum compared with that in plasma is discussed in view of the paracellular pathway.

We have modified deproteinization methods and a number of spectrophotometric and bioluminescent methods in order to measure the concentrations of cellular metabolites in small volumes (< 0·3 ml) of sows' colostrum and milk. For the majority of the assays, recoveries ranged from 92 to 105%. The binding of ATP and UTP to a calcium phosphate–citrate–caseinate complex in milk, and the decrease in ATP (92%/h) and UTP (18·1 %/h) concentrations during in vitro incubation of the whey fraction suggested that it was unlikely ATP and UTP (both < 1 µM) could exist free in sows' milk. The mean concentrations (range) of cellular metabolites in milk (6–11 d post partum) were: glucose, 669 µM (220–1367); glucose 6-phosphate, 63·0 µM (27·6–101·4); glucose 1-phosphate, 18·3 µM (13·1–24·8); UDPglucose, 296 µM (170–494); UDPgalactose, 635 µM (230–945); lactose, 162 mM (124–187); UDP, 105 µM (85–130); UMP, 1760 µM (1326–2587); inorganic phosphate, 135 mM (1·4–29·3); ATP, <0·5 µM; ADP, 53·6 µM (10·5–171·25); AMP, 215 µM (61·6–491·6); cAMP, 22·3 µM (3·5–61·6); galactose, 198 µM (118–474) and fructose, 226 µM (172–283). Differences in the concentrations of glucose, glucose 6-phosphate, glucose 1-phosphate, UDPgalactose and cAMP between fore and hind milk samples indicated postsecretory changes in the concentrations of certain metabolites. Changes in the concentrations of metabolites during in vitro incubation of milk and of colostrum suggested that these postsecretory changes were probably due to the actions of enzymes present in mammary secretion. Collection of milk that has been most recently secreted (hind milk) will provide the most accurate indication of the intracellular concentrations of these metabolites. The concentrations of cellular metabolites in the milks from different species are compared.

The concentrations of lactose, glucose, glucose 6-phosphate, glucose 1-phosphate, UDPglucose, UDPgalactose, UDP, UMP, inorganic phosphate, ADP and AMP (metabolites involved in the lactose synthesis pathway), and cAMP, galactose and sodium were measured in the mammary secretion from four or five mammary glands on each of six sows during the first 5 d post weaning. The concentrations of lactose, glucose and galactose were also measured in plasma during this time. Following weaning, the rapid increase in the concentrations of glucose 6-phosphate and UDPgalactose suggested that the rate of lactose synthesis was regulated by the inhibition of hexokinase and/or lactose synthase, while the decrease in glucose and AMP indicated a subsequent decline in glucose and ATP utilization. The rapid increase in glucose 6-phosphate which plays a pivotal role as a substrate for both lactose and de novo fatty acid synthesis, and the rapid decrease in AMP which reflects ATP utilization, were good markers of decreased metabolic activity. These rapid changes in the metabolic activity of the mammary glands were not observed in a second weaning study when two piglets were removed from selected mammary glands for periods up to 5 h during established lactation. Since concentrations of lactogenic hormones remain elevated following partial weaning, but fall following total weaning (Rojkittikhun et al. 1991), these differences in mammary gland metabolism indicate that endocrine rather than autocrine mechanisms are controlling lactose and fat synthesis during the initial stages of total weaning.

The functional significance of citrate in the mammary secretion of six sows was investigated during the second stage of lactogenesis (lactogenesis II) and established lactation. The changes in the concentrations of progesterone and lactose in the maternal blood, and lactose, Na and K in the mammary secretion, suggested that lactogenesis II began during the final day of pregnancy. The concentration of citrate in the mammary secretion of the sows during lactogenesis II was high and varied from 5·4 (SEM 0·5) mai at day 0·5 post partum to 6·8 (SEM 0·4) mM at day 1·5 post partum. There was a decline of ˜ 30% in the concentration of citrate in the milk of sows during the first week of lactation. These findings suggest that, in contrast to all other species studied previously, milk citrate is not a harbinger of lactogenesis II in the sow. However, the changes in the concentration of citrate in the mammary secretion of sows may reflect changes in the rate of de novo synthesis of fatty acids that take place in the mammary glands of sows during lactogenesis II and established lactation.

A new sampling method for the collection of fore and hind milk from the sow has been developed which resembles normal milk removal by the piglet, yet overcomes the difficulties of collecting milk that is available for only a short time (10–20 s) at each let-down. Samples of hind milk were collected immediately after the completion of a successful sucking, while the fore milk was collected at the beginning of the next natural let-down. Modification of existing assays for fat, protein and lactose was required to provide rapid analysis of the small volumes (< 0·5 ml) of milk collected using this procedure, and these methods were validated by comparison with reference methods. Total solids and the concentration of fat in whole milk, and protein and lactose in skim milk, were measured in fore and hind milk collected from 4, 20, 12 and 12 sows respectively, 6–11 d post partum. For fore milk, the results (mean ± SD (n)) were: total solids, 199·9±9·9 g/1 (8); fat, 96·7 ± 13·9 g/1 (41), protein, 47·7±4·5 g/1 (27) and lactose, 58·3 + 4±0 g/1 (27). In hind milk, there was a significant increase in the concentration of fat (15·3 g/1, p < 0·001, n = 41) which was reflected by a significant increase in total solids (14·7 g/1, P < 0·001, n = 8) and calculated energy (511 kj/l, P < 0·001, n = 27), but there was no significant difference in the concentration of either protein or lactose. This increase in milk fat during milk letdown is in contrast to the results of most previous studies in the sow and is discussed in terms of the possible mechanisms that may cause the concentration of fat to increase as milk is removed from the mammary gland.

The onset of copious milk secretion (lactogenesis II) in women occurs between 1 and 3 d after birth, and during this period the composition of breast milk changes. During the first 5 d of lactation we measured the concentrations of total, diffusible and ionized Ca (Catot, Cad, Ca2+), diffusible phosphate (Pid), diffusible citrate (Citd) and lactose in the breast milk. On day 1 after birth the concentrations (mean ± SEM) were Catot, 5·71±0·30 mM; Cad, 2·66±0·19 mM; Ca2+, 2·90 ±0·18 mM; Pid, 0·26±0·16 mM; Citd, 0·25±0·03 mai and lactose, 76±11 mM. Between day 1 and day 4 the concentration of Catot increased 1·7-fold to 9·56 ±0·39 mai, Cad increased 1·8-fold to 4·75±0·26 mM, Ca2+ decreased by 20% to 2·33 ± 0·13 mM, Pid increased 6·6-fold to 1·69±0·11 mM, Citd increased 20-fold to 5·06±0·21 mM, and lactose increased 2·3-fold to 173±4 mM. A high correlation has been found between [Cad] and [Citd] in the milk of both ruminant and non-ruminant species, which show a wide range in concentrations of [Cad] and [Citd], and the data fit a simple physicochemical model of ion equilibria in the aqueous phase of milk. The results of the present study confirm the relationship between [Cad] and [Citd] in human milk, even during lactogenesis II when the composition of the milk is changing very rapidly.

Changes in the concentrations of glucose and galactose were measured in the peripheral blood of ten piglets after they had ingested milk during a natural sucking. In addition, the mild stress associated with the experimental procedure was determined by sampling nine fasted piglets over a period of 9 to 12 min. During this period there was a significant increase in the concentration of glucose in the blood of the piglets but no change in the concentration of galactose. After milk ingestion during a natural sucking the concentrations of both glucose and galactose increased from 5·7 mM and 19 μM to reach peak values of 7·7 mM and 122 μM, respectively, by 30 to 35 min. The concentrations of glucose and galactose returned to initial values in 60–80 min and 80–100 min, respectively, after sucking. Since the change in the concentration of galactose in the peripheral blood was much lower than the change in the concentration of glucose, we conclude that galactose was rapidly removed by the livers of sucking piglets. However, after the ingestion of milk the percentage increase (from initial to peak values) in the concentration of galactose in the blood was much larger (650%) than the increase in the concentration of glucose (43%). Thus, we propose that the determination of galactose in the peripheral blood may provide a qualitative method for monitoring the digestion and absorption of milk lactose in sucking piglets.