First Few Days Of Lactation Research Articles

Effect of sialyllactose administration on growth performance and intestinal epithelium development in suckling piglets

Sialyllactose (SL) is an abundant carbohydrate in sow’s colostrum, but decreases during the first few days of lactation. Here, we investigated its effect on growth performance and intestinal epithelium development in pigs during the suckling period. Sixteen litters of suckling piglets were divided into two groups (Control and SL group). All piglets were breast fed and the SL group was orally administrated with 4 g SL per day for 21 d. Results showed that SL administration increased the litter weight (P = 0.043) and tended to elevate the average body weight (P = 0.084) at weaning. The serum concentrations of immunoglobulin G and glutathione peroxidase were elevated, but the concentrations of tumor necrosis factor α, malondialdehyde, and diamine oxidase were decreased upon SL administration (P < 0.05). Moreover, SL administration elevated the villus height, the ratio of villus height to crypt depth, and the activities of lactase and sucrase in jejunum (P < 0.05), and decreased the total apoptotic cells in ileal epithelium (P = 0.003). The expression levels of tight-junction protein ZO-1 in the jejunum and ileum, and critical functional genes such as the glucose transporter 2, divalent metal transporter 1, and insulin growth factor 1 receptor in the jejunum were elevated by SL supplementation (P < 0.05). Importantly, SL increased butyric acid concentration, and elevated the abundances of Lactobacillus and Bifidobacterium in cecum (P < 0.05). These results suggest that SL could improve the growth performance of suckling piglets, which was associated with improved immunity and antioxidant capacity, as well as the intestinal barrier functions.

Fatty Acid Profile and Enterolactone Content of Early and Commercial Milk of Dairy Cows Supplemented with Flaked Flaxseed during the Dry Period.

Simple SummarySeveral studies have been published on n-3 fatty acids enrichment of food of animal origin by the use of specific supplements in animal feeding. Flaxseed (Linum usitatissimum) is known as an excellent source of unsaturated fatty acids, particularly of alpha-linolenic acid. Moreover, most of the plant lignans contained in flaxseeds are converted by the animal in mammalian lignans, such as enterodiol and enterolactone. Thus, milk with elevated enterolactone may be an efficient strategy to optimize the effects of lignans on human health. This study aimed to investigate the effect of flacked flaxseed supplementation on dairy cow during the pre-partum period. The increase of n-3 on milk obtained from cow fed the flaxseed diet was detected only in the first days of lactation. In addition, an increase of enterolactone on milk from flaxseed fed cows was observed only at 15th day after calving; a higher amount of enetrolactone was detected in plasma and milk from the 15th day after calving, independently from the treatment group. This study suggests that the duration of n-3 carry-over into the milk is dependent on the n-3 feeding source concentration. Finally, more studies are needed to investigate enterolactone metabolism in ruminants.Various supplementations in animal feeding have been investigate in order to enrich food of animal origin with n-3 fatty acids. Although the effects of flaxseeds inclusion in diets for lactating dairy have already been assessed, few studies have focused on this n-3 source supplementation during the transition period. The aim of this work was to evaluate the effects of flacked flaxseed (200 g/head/day; 2.13% DM) dietary treatment during the dry period on milk yield and quality in the 30 days after calving. In addition, the enterolactone content in plasma (before and after calving) and in milk of cows fed diets supplemented or not with flaxseed was considered. The study demonstrated that the carry-over effect on the milk profile of C18:2, C18:3 n-3, and C20:5 n-3 was significantly higher in flaxseed diet than in the control one at 4th day of lactation. A significant increase of enterolactone on milk from flaxseed fed cows was observed only at 15 sampling day. The quick modification in fatty acid (FA) profile of the milk in the first few days of lactation suggests that the carry over effect from pre-calving flaxseed feeding at this concentration was very short lasting.

Antiviral oxysterols are present in human milk at diverse stages of lactation

Oxysterols are cholesterol oxidation derivatives. Those containing an additional hydroxyl group on the side chain of the cholesterol molecule result from a physiological enzymatic synthesis and include the majority of oxysterols present in the circulation. Among these, 25-hydroxycholesterol (25OHC) and 27-hydroxycholesterol (27OHC) are characterized by a broad antiviral activity and are now considered involved in the innate immune response against viruses. Despite the emerging role of these sterols in the innate antiviral defences, no data are available on their presence in human breast milk (BM) to date. In this study, we investigated the content of oxysterols of enzymatic synthesis in BM of twelve donor mothers at different stages of lactation (i.e. in colostrum, transitional milk, and mature milk) by gas chromatography-mass spectrometry analysis. The side-chain oxysterols 25OHC, 27OHC, and 24S-hydroxycholesterol (24SOHC) were actually present in BM in all stages of lactation, but the concentration of 27OHC showed a remarkable peak in colostrum. Antiviral assays revealed that all the colostrum samples contained 27OHC concentrations that were active in vitro against two relevant pediatric viral pathogens: the human rotavirus and the human rhinovirus. Overall, this study discloses new antiviral components of BM and suggests a passive transfer of these protective factors to the infant via breastfeeding, especially in the first few days of lactation.

Effect of inhibiting the lactogenic signal at calving on milk production and metabolic and immune perturbations in dairy cows

During the periparturient period, the abrupt increase in energy demand for milk production often induces metabolic and immunological disturbances in dairy cows. Our previous work has shown that reducing milk output by milking once a day or incompletely in the first few days of lactation reduces these disturbances. The aim of this study was to reduce metabolic and immunological disturbances by limiting milk production during the first week of lactation by inhibiting the lactogenic signal driven by prolactin. Twenty-two fresh cows received 8 i.m. injections of the prolactin-release inhibitor quinagolide (QUIN; 2 mg) or water as a control (CTL). The first injection was given just after calving, and the subsequent 7 injections were given every 12 h. Milk production was measured until d 28 after calving. Blood samples were taken from d 1 (calving) to d 5 and then on d 7, 10, 14, 21, and 28 to measure concentrations of urea, phosphorus, calcium, glucose, nonesterified fatty acids (NEFA), β-hydroxybutyrate, and prolactin. Other blood samples were taken on d 2, 5, 10, and 28 to analyze oxidative burst, phagocytosis, and the effect of the serum on the lymphoproliferation of peripheral blood mononuclear cells from donor cows. Blood prolactin concentration was lower from d 2 to 5 but higher from d 10 to 28 in the QUIN cows than in the CTL cows. Milk production was lower from d 2 to 6 in the QUIN cows than in the CTL cows (24.3 ± 6.4 and 34.8 ± 4.1 kg/d on average, respectively). We observed no residual effect of quinagolide on milk production after d 6. During the first week of lactation, blood glucose and calcium concentrations were higher and β-hydroxybutyrate concentration was lower in the QUIN cows than in the CTL cows. Blood NEFA, urea, and phosphorus concentrations were not affected by the treatment. At d 2 and 5, the phagocytosis ability of polymorphonuclear leukocytes was not affected by treatment; however, quinagolide injection enhanced the proportion of cells that entered oxidative burst, The mitogen-induced proliferation of peripheral blood mononuclear cells was greater when they were incubated with serum harvested from the CTL cows and was negatively correlated with the NEFA concentration in the serum. Reducing the prolactin peak at calving was effective in reducing milk production during the first week of lactation without compromising the dairy cow's overall productivity. Slowing the increase in milk production allowed a more gradual transition from pregnancy to lactation and led to a reduction in metabolic stress and an improvement in some immune system aspects during this period.

Relationships between rumination time, metabolic conditions, and health status in dairy cows during the transition period1

The main objective of this experiment was to monitor the rumination pattern during the transition period in primiparous (PR) and pluriparous (PL) dairy cows and to investigate its relationships with metabolic conditions, milk yield, and health status. The study was carried out in an experimental free-stall barn and involved 32 Italian Friesian cows (9 PR and 23 PL) during the transition phase. The rumination time (RT) was recorded with an automatic system (HR-Tag), and data were calculated and summarized in 2-h intervals. Blood samples were collected during the transition phase to assess biochemical variables related to energy, protein, and mineral metabolism, as well as markers of inflammatory conditions and some enzyme activity. Daily milk yield, BW, nutritional condition, and health status were also recorded. The average RT before calving (-20 to -6 d) was 463 min/d in PR (range 270 to 620) and 522 min/d in PL (range 411 to 640). In the early lactation [15 to 40 d in milk (DIM)], the average RT was 504 min/d in PR (range 400 to 585) and 562 min/d in PL (range 414 to 685) and was positively correlated with milk yield (r = 0.36; P < 0.001). The RT reached the minimum at calving d (262 min/d in PR and 278 min/d in PL). Before calving the percentage of RT during the nighttime was 60% in PR and 62% in PL, whereas the values decreased after calving (55% in PR and 57% in PL). During the first weeks of lactation, PR showed a shorter RT than PL in the 2-h intervals that included milkings. Cows with reduced RT before calving maintained reduced RT after calving and suffered a greater frequency of disease than cows with greater RT in late pregnancy. Moreover, cows characterized by mild inflammatory conditions and without health disorders or only mild health disorders during the puerperium showed a greater average rumination time (over 520 min/d) during the first 10 d of lactation. Conversely, the decreased RT (450 min/d) during the first few days of lactation was observed in cows with subclinical diseases or health disorders. Cows affected by clinical mastitis during the trial showed a reduction of RT and a change in its variability already some days before the drug treatment. Our results suggest that the automatic measurement of RT is useful to predict calving time and to quickly obtain information on health status of the animals in a period as critical as the transition phase.

Relationships between rumination time, metabolic conditions, and health status in dairy cows during the transition period

The main objective of this experiment was to monitor the rumination pattern during the transition period in primiparous (PR) and pluriparous (PL) dairy cows and to investigate its relationships with metabolic conditions, milk yield, and health status. The study was carried out in an experimental free-stall barn and involved 32 Italian Friesian cows (9 PR and 23 PL) during the transition phase. The rumination time (RT) was recorded with an automatic system (HR-Tag), and data were calculated and summarized in 2-h intervals. Blood samples were collected during the transition phase to assess biochemical variables related to energy, protein, and mineral metabolism, as well as markers of inflammatory conditions and some enzyme activity. Daily milk yield, BW, nutritional condition, and health status were also recorded. The average RT before calving (-20 to -6 d) was 463 min/d in PR (range 270 to 620) and 522 min/d in PL (range 411 to 640). In the early lactation [15 to 40 d in milk (DIM)], the average RT was 504 min/d in PR (range 400 to 585) and 562 min/d in PL (range 414 to 685) and was positively correlated with milk yield (r = 0.36; P < 0.001). The RT reached the minimum at calving d (262 min/d in PR and 278 min/d in PL). Before calving the percentage of RT during the nighttime was 60% in PR and 62% in PL, whereas the values decreased after calving (55% in PR and 57% in PL). During the first weeks of lactation, PR showed a shorter RT than PL in the 2-h intervals that included milkings. Cows with reduced RT before calving maintained reduced RT after calving and suffered a greater frequency of disease than cows with greater RT in late pregnancy. Moreover, cows characterized by mild inflammatory conditions and without health disorders or only mild health disorders during the puerperium showed a greater average rumination time (over 520 min/d) during the first 10 d of lactation. Conversely, the decreased RT (450 min/d) during the first few days of lactation was observed in cows with subclinical diseases or health disorders. Cows affected by clinical mastitis during the trial showed a reduction of RT and a change in its variability already some days before the drug treatment. Our results suggest that the automatic measurement of RT is useful to predict calving time and to quickly obtain information on health status of the animals in a period as critical as the transition phase.

Analysis of Insulin in Human Breast Milk in Mothers with Type 1 and Type 2 Diabetes Mellitus

Despite the important role that insulin plays in the human body, very little is known about its presence in human milk. Levels rapidly decrease during the first few days of lactation and then, unlike other serum proteins of similar size, achieve comparable levels to those in serum. Despite this, current guides for medical treatment suggest that insulin does not pass into milk, raising the question of where the insulin in milk originates. Five mothers without diabetes, 4 mothers with type 1, and 5 mothers with type 2 diabetes collected milk samples over a 24-hour period. Samples were analysed for total and endogenous insulin content and for c-peptide content. All of the insulin present in the milk of type 1 mothers was artificial, and c-peptide levels were 100x lower than in serum. This demonstrates that insulin is transported into human milk at comparable concentration to serum, suggesting an active transport mechanism. The role of insulin in milk is yet to be determined; however, there are a number of potential implications for the infant of the presence of artificial insulins in milk.

Hypernatremia

Hypernatremia

Evaluation of a Mechanistic Model of Glucose and Lipid Metabolism in Periparturient Cows

A mechanistic model was previously developed to quantitatively describe glucose and lipid metabolism in periparturient cows. The objectives of the current study were to evaluate the accuracy and precision of the model by comparing predictions to data collected in an independent experiment; to identify the critical metabolic processes for ketosis development; and to use the model to evaluate the relative importance of dry matter intake, calf birth weight, milk yield, and body condition score on nutrition management. Residuals (observed – predicted) were regressed on model predictions using the independent data for the model inputs, and prediction error was calculated. Each model parameter (e.g., the rate of glucose consumption by peripheral tissues) was increased independently by 1 standard deviation to identify the critical metabolic processes for ketosis development. Critical control points to prevent ketosis were identified by increasing the driving variables of the model by 1 standard deviation to estimate the response in ketone body formation. The root mean square prediction error was 0.527mM for ketone body predictions. The sensitivity analysis indicated that in the first few days of lactation, the rate of nonesterified fatty acid utilization had a greater effect on ketone body concentrations in periparturient cows than the other parameters tested in the model. The model was consistent with the knowledge that over-fattening during the prepartum period should be avoided to help prevent ketosis.

Estimation of Genetic Parameters for Perinatal Sucking Behavior of Italian Brown Swiss Calves

Brown Swiss breeders sometimes experience difficulties in feeding calves because of the weak sucking ability of the calves in the early days of life. For the welfare of the calves, they should be suckled by their dams or should aggressively ingest colostrum immediately after birth. The composition of colostrum changes rapidly during the first few days of lactation, and the ability of calves to absorb the Ig decreases quickly as well. The aim of this study was to increase our knowledge of environmental and genetic components affecting the sucking response, to evaluate the possibility of selecting for this trait. Sucking ability was recorded in 3 categories (drank from the milk bucket nipple or bottle without help, drank with help, did not drink) at 5 post-natal meals (6, 12, 24, 48, and 72h from birth). Records were analyzed with 2 different models: a single-trait threshold sire model, in which all observations were analyzed as a single trait with 5 levels, and a multiple-trait threshold liability sire model, in which meal-by-meal observations were analyzed as 5 different binary traits. Management procedures, the interval between birth and meals, parity, and season of birth were environmental factors affecting the variability in sucking ability. The heritability estimate for the single-trait analysis was 0.14, whereas heritabilities for the multiple-trait analysis were 0.26, 0.22, 0.21 0.12, and 0.13 for the first, second, third, fourth, and fifth meal, respectively. Estimated genetic correlations among traits were high (0.82 to 0.99). This study suggests the possibility of selection based on sucking ability. Future collection of larger data sets on the sucking response of calves in the first 2 meals after birth would increase the accuracy of genetic parameter estimates.

Milk L-lactate concentration is increased during mastitis.

A study was undertaken in cattle to evaluate changes in milk L-lactate in relation to mastitis. A healthy, rear quarter of the udder of each of ten cows in mid-lactation was infused with 1000 colony-forming units (cfu) of Streptococcus uberis following an afternoon milking. Foremilk samples were taken at each milking from control and treated quarters and antibiotic treatment was applied following the onset of clinical mastitis or after 72 h. One cow did not become infected. Six quarters showed clinical symptoms of mastitis within 24-40 h and this was associated with a more than 30-fold increase in milk L-lactate (to 3.3 mM) and an increase in somatic cell count (SCC) from 4.5 x 10(3) to 1 x 10(7) cells/ml. Three cows were subclinical, with cell counts ranging from 1.5 x 10(6) to 1 x 10(7) cells/ml. In these animals, milk lactate ranged from 0.7 to 1.5 mM in the infected quarters up to 40 h post-infection, compared with less than 0.1 mM in control quarters. Milk was examined from 137 cows in mid-lactation which were known to have mastitis. Foremilk samples were taken aseptically from control and infected quarters of cows on commercial farms. Mean milk L-lactate concentrations and SCC were 0.14 +/- 0.02 mM and 1.85 +/- 0.3 x 10(5) cells/ml, respectively, in control (bacteriologically negative) samples. However, L-lactate concentrations exceeded 2.5 mM in the presence of some types of infection, the level of the lactate response being closely related to the impact of the infection on SCC. L-Lactate concentrations were relatively elevated in milk samples taken post partum, declining from 0.8 to 0.14 mM oyer the first few days of lactation. In conclusion, milk L-lactate has potential as an indicator of clinical and subclinical mastitis in dairy cows.

Prepartum teat apex colonization with Staphylococcus chromogenes in dairy heifers is associated with low somatic cell count in early lactation

A high number of dairy heifers freshen with udder health problems. The prevalence of teat apex colonization (TAC) with Staphylococcus chromogenes, one of the most widespread coagulase-negative staphylococci (CNS) in milk samples from freshly calved dairy heifers, was measured cross-sectionally in non-lactating heifers on eight commercial dairy farms in Belgium. The influence of age on this prevalence, and the association between teat apex colonization with S. chromogenes prepartum and quarter milk somatic cell count (SCC) in early lactation were studied. In total, 492 teat apices were sampled from 123 heifers. The age of the heifers varied from 8 to 34 months. Overall, 20% of the heifers had at least one teat apex colonized with S. chromogenes. Of all teats sampled, 10% were colonized with S. chromogenes. The chance of having at least one teat apex colonized with S. chromogenes increased with age of the heifer. The presence of prepartum teat apex colonization with S. chromogenes was not associated with intramammary infection (IMI) early postpartum with the same bacterium. On the contrary, teat apex colonization with S. chromogenes prepartum appeared to protect quarters in the first few days of lactation from having somatic cell count ≥200,000 cells/ml milk, commonly accepted as the threshold for intramammary infection.

Expression of cysteine sulfinate decarboxylase mRNA in rat mammary gland.

Our previous report demonstrated that high concentration of taurine is present in rat milk for the first few days of lactation and plays an important role in the body growth of rat pups. In the present study, gene expression of rate-limiting enzyme for taurine biosynthesis, cysteine sulfinate decarboxylase (CSD) were examined in rat mammary gland. By RT-PCR, CSD mRNA was found to be expressed in rat mammary gland like that in the liver. The expression level of CSD mRNA in the mammary gland was higher in the earlier lactational stage (days 1 and 6 of lactation) than that in the later lactational stage (day 14). CSD mRNA expression in the mammary gland of non-pregnant rats was only a trace level. By in situ hybridization analysis, CSD mRNA was demonstrated in the epithelial cells of the mammary gland. These results suggest that high concentrations of taurine in the milk are at least partially resulted from de novo synthesis of taurine in mammary gland epithelial cells and that the expression pattern of CSD mRNA may be responsible for the changes in taurine levels in the milk during a lactational period.

Lactotrope differentiation in rats is modulated by a milk-borne signal transferred to the neonatal circulation.

We have previously reported that normal differentiation of PRL-secreting cells in neonatal rats requires a maternal signal specific to the first few days of lactation. Subsequently, we found that milk proteins from this period had a greater capacity than those from later in lactation to stimulate lactotrope differentiation in vitro, suggesting that the maternal signal was a milk-borne peptide. We reasoned in the present study that if this were the case, the signal should be present in the serum of suckling pups, and its level should reflect that in mothers' milk. To test this hypothesis, we first compared serum from 3-day-old pups with that from newborn pups that had not yet suckled. Anterior pituitary cells from 1-day-old pups were cultured for 6 days in the presence of various serum samples (0.01-1.0% by volume). Lactotrope differentiation was assessed using a reverse hemolytic plaque assay for PRL. Serum from 3-day-old pups was more effective (P < 0.01; n = 4) than that from unsuckled newborns at stimulating lactotrope differentiation in vitro (to 3.5 +/- 0.5% and 1.5 +/- 0.5% of all anterior pituitary cells, respectively). Next, we tested whether this increase in serum bioactivity in 3-day-old pups required maternal influences restricted to the first few days of lactation. Newborn pups were placed with foster mothers that had been lactating for 0 or 7 days. When the pups were 3 days old, their serum was tested for lactotrope-differentiating activity. Serum from pups placed with day 0 foster mothers (control) increased the abundance of PRL cells to 2.4 +/- 0.3% of all cells (P < 0.01; n = 3), whereas serum from pups placed with day 7 foster mothers failed to stimulate lactotrope differentiation. In similar experiments, litters placed the day after birth with day 7 foster mothers were separated on day 3 postpartum for 3 h, after which littermates were allowed to suckle for 90 min on different foster mothers on day 3 or 9 of lactation. This brief suckling bout increased (P < 0.01; n = 3) the capacity of serum from pups on day 3 mothers to stimulate PRL cell differentiation (to 3.9 +/- 0.4%) above that of serum from littermates placed with day 9 foster mothers (to 1.8 +/- 0.2%). Finally, the differentiating activities in pup serum and mothers' milk eluted from gel filtration chromatography in identical fractions (4-8 kilodaltons).(ABSTRACT TRUNCATED AT 400 WORDS)

Tumor necrosis factor-alpha in human milk.

We previously demonstrated that certain biologic activities in human milk were partially blocked by antibodies directed against human tumor necrosis factor-alpha (TNF-alpha). In this study, immunochemical methods were used to verify the presence of TNF-alpha in human milk obtained during the first few days of lactation. Gel filtration revealed the presence of TNF-alpha by RIA in molecular weight fractions between 80 and 195 kD. TNF-alpha could not be detected consistently by conventional Western blotting or cytotoxic assays. Although immunoreactive bands were detected by a Western blot-125I protein A technique in TNF-alpha-positive fractions from gel filtration, those bands proved to be nonspecific. TNF-alpha in milk was reliably quantified by the competitive RIA. Those studies revealed that the concentrations of TNF-alpha in milk were 620 +/- 183 pg/mL. Although RNA to TNF-alpha was detected in milk leukocytes by Northern blotting, little TNF-alpha was found in those cells before or after stimulation with N-formyl-l-methionyl-l-leucyl-l-phenylalanine or 4 beta-phorbol-12 beta-myristate-13 alpha-acetate. The origin of this cytokine in human milk remains unclear. Nevertheless, this study suggests that TNF-alpha is present in early human milk in sufficient quantities to exert possible biologic effects upon the mammary gland of the mother or the immune system of the infant.

Stimulation of Prolactin Cell Differentiationin Vitroby a Milk-Borne Peptide*

We have previously reported that the normal expression of PRL-secreting cells in neonatal rats requires a maternal signal specific to the first few days of lactation. These results raised the possibility that a milk-borne factor(s) ingested by the neonate and absorbed into the circulation might induce the ontogenic appearance of PRL cells. The purpose of the present study was to determine whether milk from this period could directly stimulate the differentiation of PRL secretors in culture. Monodispersed anterior pituitary cells from 1-day-old pups were cultured for 6 days with aqueous extracts of milk from early (days 2, 3, and 4) and late (days 15 and 16) lactation and then subjected to reverse hemolytic plaque assays for PRL and GH release. We found that the addition of milk extracts (10 mg/ml) from either early or late lactation stimulated the differentiation of PRL secretors (to 6.1 +/- 1.0% and 2.4 +/- 0.7% of all pituitary cells, respectively; mean +/- SE; n = 3) above that in control cultures without milk (0.2 +/- 0.2%). Thus, early milk was more than twice as effective as late milk in this regard (P less than 0.05). This effect appeared to be specific to PRL cell differentiation, since the relative abundance of GH secretors was not different between cells treated with either early or late milk (29.3 +/- 4.8% and 33.7 +/- 3.9%, respectively). On the other hand, late milk was more than twice as effective as early milk at increasing the capacity of GH secretors to release hormone (P less than 0.05). Preliminary characterization by gel filtration chromatography and proteolytic hydrolysis indicates that the bioactivity that differentiates PRL secretors is a small peptide(s) of 2000-6000 daltons. Taken together, our results demonstrate that a milk-borne peptide(s) is capable of specifically stimulating the differentiation of PRL-secreting cells in vitro, and that this bioactivity is more prevalent in milk from early lactation.

Cholesterol metabolism in the rat lactating mammary gland: The role of cholesteryl ester hydrolase

An acid cholesteryl ester hydrolase activity associated with a fraction containing mitochondria and lysosomes from rat lactating mammary glands was found to have a pH optimum of 5.0. Its sedimentation pattern was closely related to that of the lysosomal enzyme markers acid phosphatase and beta-glucuronidase, suggesting that the activity is associated with the lysosomes. The enzyme was strongly inhibited by Cu2+, but was inhibited little by other divalent metal ions. Acid cholesteryl ester hydrolase activity was almost completely abolished by p-hydroxy-mercuribenzoate, but this effect was reversed in the presence of an equimolar concentration of reduced glutathione (GSH), indicating that the enzyme requires free sulfhydryl groups for activity. These properties are similar to those of acid, lysosomal cholesteryl ester hydrolases found in other tissues. Acid cholesteryl ester hydrolase activity was 8-14 fold higher in mammary tissue from lactating as compared to virgin rats. Neutral cholesteryl ester hydrolase activities associated with the microsomal and cytosolic subcellular fractions were also increased in lactating glands, but to a lesser extent. In addition, a 2-fold increase in the activities of both the acid and microsomal neutral enzymes was seen during the first few days of lactation, while the cytosolic neutral activity remained constant. These results suggest that mammary gland cholesteryl ester hydrolases have a role in the regulation of cholesterol metabolism in mammary cells, and in the provision of cholesterol for secretion into milk.

Normal Differentiation of Prolactin Cells in Neonatal Rats Requires a Maternal Signal Specific to Early Lactation*

This laboratory has previously demonstrated that PRL-secreting cells are virtually nonexistent on day 3 but appear in appreciable numbers on day 4 of neonatal life. The purpose of the present study was to determine whether this explosive appearance of PRL cells is due to maternal influences specific to the first 4 days of lactation. Litters of 1-day-old rat pups were placed with foster mothers that had been lactating for either 1 or 4 days. Four days later (at 5 days of age), the anterior pituitaries from these pups were removed, dispersed into individual cells with trypsin, and subjected to reverse hemolytic plaque assays for PRL and GH release. We found that the proportion of PRL-releasing cells in pituitaries from pups placed with day 1 mothers (4.2 +/- 0.6%; mean +/- SE; n = 4 experiments) was similar to that in 5-day-old pups that had not been fostered (3.4 +/- 0.6%; n = 3 experiments). In contrast, placing 1-day-old pups with day 4 mothers significantly reduced (P less than 0.01) the fraction that released PRL to 0.6 +/- 0.2% of all pituitary cells (n = 4 experiments). This effect appeared to be specific to PRL cells, since the percentage of all pituitary cells that secreted GH was not different between litters fostered onto day 1 or day 4 dams (40.0 +/- 3.8% and 41.1 +/- 3.9%, respectively; n = 3 experiments). Furthermore, the suppression of PRL cell expression did not result from a gross nutritional deficit, since the rates of body wt gain during the foster period were not different between the two groups (12.4 +/- 0.5 and 12.9 +/- 0.8 g/litter.day, respectively; n = 4 experiments). In a second design, newborn pups were placed with females that had been lactating for 0, 2, 4, or 7 days and analyzed for PRL- and GH-releasing cells at 5 days of age. In two trials of this experiment, the fractions of pituitary cells that secreted PRL were 5.2, 6.3, 5.9, and 1.2%, respectively for trial 1 and 3.3, 4.1, 0.7, and 0.6%, respectively for trial 2. As in the first experiment, the proportion of GH-releasing cells and the rate of growth were not different among the four groups. Taken together, these results indicate that normal differentiation of PRL cells in neonatal rats is triggered by a maternal signal present during the first few days of lactation, and that the magnitude of this signal declines with the progression of lactation.

Changes in maternal taurine levels in response to pregnancy and lactation

Taurine levels in various tissues and fluids of female rats were measured throughout pregnancy and lactation. The taurine concentration of liver markedly increased at days 19 and 21 of pregnancy to 188% of levels for nonpregnant, nonlactating control rats and then fell rapidly after delivery to reach only 30% of the control level by 3 days post-partum. Muscle and heart taurine concentrations were significantly negatively correlated with liver taurine levels. Brain taurine levels were low at days 14, 19 and 21 of pregnancy and day 14 of lactation. Urinary excretion of taurine decreased to 32% of control levels at day 21 of pregnancy and was negatively correlated with the hepatic taurine concentration over the course of pregnancy and lactation. The ratio of glycine- to taurine-conjugated bile acids was strongly negatively correlated with the hepatic taurine concentration. The milk taurine level was positively correlated with hepatic taurine concentration during lactation. The hepatic taurine pool appears to increase just before parturition and to rapidly decrease during the first few days of lactation when high levels of taurine are secreted in the milk. Our data suggests that the accumulation of taurine in the liver may be related to both a decreased renal clearance of taurine and a shifting of tauring from other tissues to the liver and that this enlarged pool of hepatic taurine may serve as a source of taurine for secretion in the early milk.

Mammary development in mice: effects of hemihysterectomy in pregnancy and of litter size post partum.

1. Mice were hemihysterectomized on day 8 of pregnancy to reduce the number of feto-placental units.2. Fetal mortality was not affected by hemihysterectomy; mean single-pup weight at birth was increased when compared with sham-operated controls.3. Pregnant sham-operated and hemihysterectomized animals were killed on days 13 and 18 of gestation, and their mammary glands were analysed for total DNA (DNA(t)) and RNA (RNA(t)). Both were significantly lower in the hemihysterectomized group on day 18, but not on day 13.4 Milk yield was assessed, by daily weighing of the litter, in groups of sham-operated and hemihysterectomized lactating animals suckling nine pups each. There was no difference in yield between the two groups.5. One group of sham-operated mice suckling nine pups, one of hemihysterectomized mice suckling nine pups and one of hemihysterectomized mice suckling four pups were killed on day 5 of lactation for mammary gland analysis. There was no significant difference in mammary weight or DNA(t) between the sham-operated and hemihysterectomized animals suckling nine pups, although RNA(t) was still reduced in the latter. Mammary weight, DNA(t) and RNA(t) were all significantly lower in the hemihysterectomized group suckling four pups than in either of the other two groups.6. It is concluded that the less well developed mammary glands of mice which give birth to small litters are capable of compensatory growth during the first few days of lactation if a sufficiently strong suckling stimulus is given.7. It is suggested that control of mammary development by the fetus during gestation and by the suckling young during early lactation are both mechanisms designed to ensure that milk yield is appropriate to the needs of the young.