Plasma Lactose Concentration Research Articles

Taurine alleviates heat stress-induced mammary inflammation and impairment of mammary epithelial integrity via the ERK1/2-MLCK signaling pathway

Heat stress leads to milk production losses and mammary gland inflammation, which may be associated with mammary epithelium damage. Taurine is one of the most abundant free amino acids in mammals which has anti-inflammatory properties. This study aimed to explore the effect of taurine pretreatment on heat stress-induced mammary epithelial integrity disruption and inflammatory damage. In our first experiment on dairy cows our results showed that compared with animals under autumn thermoneutral condition (THI = 62.99 ± 0.71), summer heat stress (THI = 78.01 ± 0.39) significantly reduced milk yield and disrupted mammary epithelial integrity as revealed by increased concentrations of serotonin and lactose in plasma, and increased levels of SA and Na+/K+ in milk. In our second study, 36 lactating mice were randomly divided into three groups (n = 12) for a 9d experiment using a climate chamber to establish a heat stress model. Our findings suggest taurine pretreatment could attenuate heat stress-induced mammary histopathological impairment, inflammation response, and enhance mammary epithelium integrity, which was mainly achieved by promoting the secretion of ZO-1, Occludin, and Claudin-3 through inhibiting activation of the ERK1/2-MLCK signaling pathway in the mammary gland. Overall, our findings indicated that heat stress induced mammary epithelium dysfunction in dairy cows, and emphasized the protective effect of taurine on mammary health under heat stress conditions using a mouse model, which may be achieved by alleviating the mammary epithelium integrity damage and inflammation response.

Response of lactating dairy cows fed different supplemental zinc sources with and without evaporative cooling to intramammary lipopolysaccharide infusion: intake, milk yield and composition, and hematologic profile1.

The objective of this study was to determine the effect of dietary supplemental Zn source and evaporative cooling on intake, milk yield and composition, and the rate of leukocyte migration into the mammary gland following intramammary lipopolysaccharide (LPS) infusion. Multiparous Holstein cows (n = 72) were assigned to one of four treatments with a 2×2 factorial arrangement including two sources of supplemental Zn: 75 mg/kg Zn hydroxychloride or 35 mg/kg Zn hydroxychloride + 40 mg/kg Zn-Met complex (ZMC) each with or without evaporative cooling. The cooling system was implemented by the use of fans and misters over the freestall and feeding areas. On day 34 of the experiment, cows (n = 16; days in milk = 263 ± 63 d) received an infusion of 10 μg of LPS, or a saline control, in the left or right rear quarters. Individual milk samples from both quarters were collected at -12, -4, 0, 6, 12, 24, 48, 72, 96, 120, 144, and 168 h relative to infusion and analyzed for composition and bovine serum albumin. Rectal temperature and respiration rate were assessed and blood samples were collected at the same time points (with an additional sample at 3 h) for analyses of lactose and cortisol. Complete blood counts were performed on samples collected within the first 24 h post infusion. Intramammary LPS infusion reduced (P < 0.01) milk yield, DMI and feed efficiency regardless of dietary or cooling treatments. Non-cooled cows tended (P = 0.09) to have greater feed efficiency (=milk yield/DMI) at 1 d after infusion than those subjected to cooling. Intramammary LPS infusion dramatically increased (P < 0.01) milk somatic cell count (SCC) but treatments had no apparent impact on milk SCC. Compared with cooled cows, non-cooled cows had greater (P < 0.05) plasma lactose concentrations, but lower (P < 0.03) blood concentrations of neutrophils and lymphocytes at 3 h post infusion. This suggests a greater leukocyte migration into the mammary gland of heat-stressed cows. In conclusion, noncooled cows tended to maintain greater feed efficiency and appeared to have greater leukocyte migration into the mammary gland immediately after intramammary LPS infusion compared with cooled cows. Dietary supplemental Zn source had no impact on measures assessed after intramammary LPS infusion.

Low‐quality rice straw forage increases the permeability of mammary epithelial tight junctions in lactating dairy cows

It is known that milking frequency and plasma hormones play important roles in regulating mammary permeability. However, it is still not known whether nutritional factors can influence udder permeability. This study was conducted to investigate mammary epithelial tight-junction permeability in lactating dairy cows fed different forage-based diets. Twenty mid-lactating dairy cows were allocated to ten blocks based on their parity and milk yield and then randomly assigned into rice straw-based diet and alfalfa-based diet groups. Both diets contained 15% corn silage and 55% concentrate (dry matter basis). In terms of forage sources, rice straw-based diets (RS) contained 30% rice straw, whereas alfalfa-based diets (AH) contained 23% alfalfa hay plus 7% Chinese wild rye hay. The concentrations of Na+ , Na+ /K+ ratio, bovine serum albumin, and plasmin in the milk, the plasma lactose concentration, and the mRNA abundance of BCL2 associated agonist of cell death, phosphatase and tensin homolog, and insulin like growth factor binding protein 5 in the mammary gland were greater in RS-fed cows than in AH-fed animals. Mammary expression of proliferating cell nuclear antigen and occludin was lower in RS-fed cows compared with the AH-fed group. The expressions of growth hormone receptor, claudin-1, -3, -4, and ZO-1 were similar in the two diet groups. The cows fed RS showed higher mammary alveolar permeability, likely due to its effect on proliferation/apoptosis rates of mammary epithelial cells. © 2018 Society of Chemical Industry.

Adaptation of dairy cows to increasing degrees of incomplete milk removal during a single milking interval

Milk accumulation in the udder decreases milk secretion and this effect is explained as well by the effects of the quantity of milk stored in the udder as by the duration and repetition of periods of milk stasis. This experiment aimed to better understand the underlying mechanisms of decreased milk yield in response to the specific effects of the quantity of milk stored in the udder, independent from storage duration, on milk yield and composition. Sixteen Holstein cows were assigned to 4 blocks of 4 cows in a 4 × 4 Latin square design using 7-d periods, with a 4-d sampling period and a 3-d washout period. Cows were milked twice daily at approximately 0700 and 1630 h throughout the trial. Treatments consisted of 4 degrees of milk removal (100, 70, 40, and 0%) applied at one morning milking, designated M0. Effects of the quantity accumulated were studied in relation to udder distension, via measurements of the total distance between the ends of the 4 teats, and cisternal capacity, via the evaluation of cisternal area by ultrasonographic scan at 1 and 9 h after M0. The effect of the quantity accumulated was also evaluated in relation to mammary epithelium permeability by determining plasma lactose concentrations 1 h before and 4, 7, and 10 h after M0. Leaving milk in the udder at M0 decreased milk production during the M0-M1 interval in a negative curvilinear manner. As a result, M0+M1 milk yield decreased or tended to decrease significantly by -1.3, -5.3, and -12.8 kg for the 70, 40, and 0% treatments compared with the 100% treatment (41.7 ± 1.26 kg/d), respectively. Negative carry-over effects on milk yield were observed until the M3 milking only for the 40 and 0% treatments, and no differences were observed between the effects of these treatments. The total distance between teats increased significantly but to decreasing degrees during the M0-M1 interval. For the 40 and 0% treatments, cisternal area, which was increased 1 h after M0 milking, exhibited no further increase during the M0-M1 interval, suggesting cisternal distension was close to maximum. Simultaneously, lactose concentrations increased in blood plasma for only these 2 treatments, and this increase occurred earlier for the 0% treatment. It was also observed that cows presenting the earliest increases in plasma lactose concentrations during milk accumulation lost more milk in response to extended milking intervals.

Oxytocin Induces Mammary Epithelium Disruption and Could Stimulate Epithelial Cell Exfoliation.

Mammary epithelial cells (MEC) are exfoliated from the epithelium into milk, influencing the number of MEC present in the udder. This process is associated with epithelium integrity. The release of oxytocin (OT) induced by milking causes myoepithelial cell contraction, which, in turn, may stimulate MEC exfoliation through mechanical forces. To investigate the role of OT in MEC exfoliation, we inhibited or induced myoepithelial cell contraction by injecting the OT receptor antagonist atosiban (Ato) or a supraphysiological dose of OT, respectively. Eight cows were assigned to 2 treatments during 2 milkings according to a crossover experimental design: Control+OT (cows were first milked to collect standard milk and then received 5IU of OT to collect residual milk through a second milking) and Ato + OT (cows were injected with Ato (50μg/kg of body weight) and milked to collect cisternal milk, then received 5IU of OT to collect alveolar milk through a second milking). Milk MEC were purified to determine their concentration and number in milk. Mammary epithelium integrity was assessed by measuring the kinetics of plasma lactose concentration. Inhibiting myoepithelial cell contraction by Ato injection decreased the number of exfoliated MEC in milk. In contrast, OT injection increased the concentration of MEC in the residual milk and the number of MEC in the alveolar milk. Ato injection reduced plasma lactose concentration, whereas, in both treatments, OT injections increased it. Our results suggested that myoepithelial cell contraction caused by OT could stimulate MEC exfoliation into milk and was associated with epithelium disruption.

Effects of heat stress and dietary zinc source on performance and mammary epithelial integrity of lactating dairy cows

Dietary Zn and heat stress alter gut integrity in monogastric animals. However, effects of Zn on mammary epithelial integrity in heat-stressed lactating dairy cows have not been studied. Multiparous lactating Holstein cows (n = 72) were randomly assigned to 1 of 4 treatments with a 2 × 2 factorial arrangement to study the effects of environment and Zn source on performance and mammary epithelial integrity. Treatments included 2 environments [cooled (CL) or not cooled (NC)] and 2 Zn sources [75 mg/kg of supplemental Zn as Zn hydroxychloride (IOZ) or 35 mg/kg of Zn hydroxychloride + 40 mg/kg of Zn-Met complex (ZMC)]. The experiment was divided into baseline and environmental challenge phases of 84 d each. All cows were cooled during the baseline phase (temperature-humidity index = 72.5), whereas NC cows were not cooled during environmental challenge (temperature-humidity index = 77.7). Mammary biopsies were collected on d 7 and 56 relative to the onset of environmental challenge to analyze gene expression of claudin 1, 4, and 8, zonula occludens 1, 2, and 3, occludin, and E-cadherin and protein expression of occludin and E-cadherin. Deprivation of cooling increased respiration rate (64.8 vs. 73.9 breaths/min) and vaginal temperature (39.03 vs. 39.94°C) and decreased dry matter intake (26.7 vs. 21.6 kg/d). Energy-corrected milk yield decreased for NC cows relative to CL cows (24.5 vs. 34.1 kg/d). An interaction between environment and Zn source occurred for milk fat content as CL cows fed ZMC had lower milk fat percentage than other groups. Relative to CL cows, NC cows had lower concentrations of lactose (4.69 vs. 4.56%) and solids-not-fat (8.46 vs. 8.32%) but a higher concentration of milk urea nitrogen (9.07 vs. 11.02 mg/mL). Compared with IOZ, cows fed ZMC had lower plasma lactose concentration during baseline and tended to have lower plasma lactose concentration during environmental challenge. Plasma lactose concentration tended to increase at 3, 5, and 41 d after the onset of environmental challenge in NC cows relative to CL cows. Treatment had no effect on milk BSA concentration. Cows fed ZMC tended to have higher gene expression of E-cadherin relative to IOZ. Compared with CL, NC cows had increased gene expression of occludin and E-cadherin and tended to have increased claudin 1 and zonula occludens 1 and 2 gene expression in the mammary gland. Protein expression of occludin and E-cadherin was unchanged. In conclusion, removing active cooling impairs lactation performance and affects gene expression of proteins involved in the mammary epithelial barrier, and feeding a portion of dietary zinc as ZMC improves the integrity of the mammary epithelium.

Mammary epithelium disruption and mammary epithelial cell exfoliation during milking in dairy cows

The presence of mammary epithelial cells (MEC) in the milk of ruminants indicates that some MEC are shed from the mammary epithelium; however, the mechanisms that regulate the MEC exfoliation process are not known. Through the release of oxytocin, prolactin, and cortisol and through oxytocin-induced mechanical forces on the mammary epithelium, milking could participate in regulating the MEC exfoliation process. The aims of the present study were to determine the rate of MEC exfoliation throughout milking and to investigate its relationship to mammary epithelium integrity and milking-induced hormone release. Milk samples from 9 Holstein dairy cows producing 40.6 ± 1.36 kg of milk/d were collected at the beginning (after 1 and 2 min), in the middle, and at the end of milking. Milk MEC were purified using an immunomagnetic method. Blood samples were collected before, during, and after milking, and the oxytocin, prolactin, and cortisol concentrations in the samples were measured. Tight junction opening was assessed by plasma lactose concentration and the Na+:K+ ratio in milk. The somatic cell count in milk varied during the course of milking; it decreased at the beginning of milking and then increased, reaching the highest values at the end of milking. Exfoliated MEC were present in all milk samples collected. The presence of MEC in the milk sample collected during min 1 of milking, likely corresponding to the cisternal milk fraction, suggests that MEC were exfoliated between milkings. The observed increase in the Na+:K+ ratio in milk and in the plasma concentration of lactose indicated that disruption of mammary epithelium integrity occurred during milking. The MEC exfoliation rate at milking was not correlated with the variables describing milking-induced prolactin release but was negatively correlated with cortisol release, suggesting that cortisol may play a role in limiting exfoliation. In conclusion, milking induced a disruption of the mammary epithelial barrier. Mammary epithelial cells may be continuously exfoliated between milkings or exfoliated during milking as a consequence of the oxytocin-induced mechanical forces and the disruption of mammary epithelium integrity.

345 Feed restriction increases mammary epithelial cell exfoliation rate in dairy cows

In dairy cows, feed restriction is known to decrease milk yield by reducing the number of mammary epithelial cells (MEC) in the udder, due to an increased rate of MEC apoptosis. The exfoliation of MEC from the mammary epithelium into milk is another process that could participate in regulating MEC number. We hypothesized that the decreased number of MEC in the udder of feed-restricted cows might also result from an increase in MEC exfoliation rate. Nineteen Holstein dairy cows producing 40 ± 0.7 kg at 80 ± 4 DIM were divided into 2 groups, control (n = 9) and feed restricted (n = 10). Ad libitum DMI was recorded during a pre-experimental period of 2 wk. For 29 d (period 1), cows were fed either 100 or 80% of their DMI measured during the pre-experimental period. Then, all cows were fed ad libitum for 35 d (period 2). Feed intake and milk production were recorded daily. Blood and milk samples were collected once during the pre-experimental period and 5, 9, and 27 d after the beginning of periods 1 and 2. Mammary epithelial cells were purified from milk samples using an immunomagnetic method. Lipid mobilization was assessed by plasma NEFA concentration. Mammary epithelium integrity was assessed by plasma lactose concentration, milk Na, and milk Na:K ratio. Mammary tissue samples were collected by biopsy at the end of each period in order to analyze the rate of cell proliferation and apoptosis. Data were analyzed using the mixed procedure of SAS. Compared with control feeding, feed restriction induced a decrease in DMI by 20% (P < 0.001), an increase in plasma concentration of NEFA (P < 0.001), and a 3-kg/d milk yield loss (P < 0.001) but had no effect on the rate of proliferation and apoptosis in the mammary tissue. Daily MEC exfoliation rate was 65% greater in feed-restricted cows compared with control cows (P < 0.05). Feed restriction had no effect on plasma lactose concentration and milk Na:K ratio but increased milk Na concentration, which does not allow conclusions about the integrity of the mammary epithelium. In period 2, when all cows returned to ad libitum feeding, no significant difference was observed on DMI, milk yield, MEC exfoliation rate, and mammary epithelium integrity between feed-restricted and control cows. These results show that the exfoliation process could play a role in regulating the number of MEC in the udder of dairy cows during a feed restriction without a carryover effect on their milk production.

Lactose in blood plasma and the ability of dairy cows to tolerate once-daily milking in terms of milk loss and milk recovery

This experiment described the variability among cows with the aim of studying the ability of dairy cows to tolerate once-daily milking (ODM) in terms of milk losses and milk recoveries observed when cows are switched from twice- to once-daily milking and then back to twice-daily milking (TDM). It also aimed to investigate whether or not lactose in blood plasma, which indicates the mammary epithelium permeability, is correlated with milk losses and recoveries and, consequently, a potential candidate assessor of dairy cow tolerance to ODM. The study used 86 crossbred dairy cows (Holstein × Normande) split into 5 groups over 2 yr. The trial consisted of 3 successive periods: a 1-wk control period of TDM, then 3 wk of ODM, followed by 2 wk of TDM. Blood samples were collected 1 to 1.5h before the morning milking and 5 to 6h after milking on d −3, 0, 1, 3, 7, 21, 23, and 28, where d 0 is the last d of the control period. Milk losses measured as kilograms per day were higher in cows with highest control-period milk yields (r=−0.66). When expressed in relative terms (%), milk losses were weakly correlated with the control-period milk yield (r=−0.22). When switched back to TDM, cows recovered only 47% of the milk lost. Milk recovery (kg/d) was weakly correlated with control milk yield (r=0.34) and not correlated with ODM milk yield. Milk recovery was correlated with milk yield losses: the higher the milk yield losses (kg/d or %), the higher the milk recovery (kg/d; r=−0.59 and −0.52, respectively). Rate of milk recovery expressed as the quantity of milk recovered per kilogram of milk lost, was not correlated with milk loss expressed as a percentage. This means that to be considered well-adapted to ODM scheduling, a cow must be well adapted to milk loss, and be well adapted to milk recovery. Blood plasma lactose concentration (log10-transformed data) was not correlated with milk production levels in either control, ODM, or back-to-TDM periods. It was poorly correlated with milk loss but was positively correlated with milk recovery (kg/d) and rate of milk recovery. We conclude that blood plasma lactose needs to be coupled with other indicators for evaluating dairy cow tolerance to ODM.

Altrenogest treatment during late pregnancy did not reduce colostrum yield in primiparous sows1

The decrease in circulating concentrations of progesterone is the lactogenic trigger in many species. The aim of the present study was to determine the effect of an orally active progestogen, altrenogest, administered in late gestation, on lactogenesis in sows. Gilts were treated with altrenogest (20 mg/d) from d 109 to 112 of gestation (ALT112, n = 6) or d 113 (ALT113, n = 8) or were not treated (control, n = 9). Colostrum production, estimated from the BW gains of the piglets, was measured during 24 h starting at the onset of parturition. Colostrum samples were collected at the onset of parturition until 48 h later. Jugular blood samples were taken from d -8 prepartum until d 3 postpartum. Altrenogest treatment extended the gestation length of ALT113 sows in comparison with control sows (116.3 vs. 114.7 d; P < 0.05). Litter size and litter weight at birth did not differ between groups (P > 0.1). Estimated colostrum yield was not reduced in altrenogest-treated sows compared with control sows (4.20 kg) and tended to be greater in ALT112 (4.73 kg) than in ALT113 sows (3.74 kg; P = 0.09). Altrenogest reduced endogenous progesterone concentrations during the 2 d prepartum in ALT113 relative to control sows (P < 0.05), likely because luteolysis occurred earlier in relation to parturition in ALT113 sows. Altrenogest reduced estradiol-17beta concentrations during the 2 d prepartum in ALT113 (P < 0.05) and ALT112 (P < 0.1) sows. Altrenogest treatment did not influence the timing of the prepartum peak of prolactin in relation to parturition. The ALT113 sows had lesser (P < 0.05) concentrations of lactose in plasma and a lesser Na:K ratio in colostrum after parturition than Control and ALT112 sows, indicating that the junctions between their mammary epithelial cells were tighter. Concentrations of colostral IgG in sows that received altrenogest tended to be less than in control sows (P = 0.08). In conclusion, altrenogest administered from d 109 to 112 or 113 of pregnancy did not affect lactogenesis in sows, possibly because the treatment delayed farrowing and main hormonal changes without affecting the relative chronology of these changes.

Short communication: Effect of estrogen supplemented at dry-off on temporal changes in concentrations of lactose in blood plasma of Holstein cows

The objective was to determine the effect of supplemental estrogen (estradiol cypionate, ECP) at dry-off on temporal changes in concentrations of lactose in blood plasma of Holstein cows as an indicator of rate of mammary involution. Thirty-two Holstein cows (8/group) were assigned randomly to 4 treatment groups: 30-d dry, 30-d dry + ECP, 60-d dry, and 60-d dry + ECP. A single injection (7.5mL) of cottonseed oil (30- and 60-d dry) or ECP (15mg) in oil (30- and 60-d dry + ECP) was administered intramuscularly at dry-off. Blood samples were collected from the coccygeal vein of all cows 24h before dry-off and at dry-off, and then 8 samples were collected throughout the subsequent 48h to monitor concentrations of lactose in blood plasma. No significant effects of ECP on the overall mean concentrations of lactose were detected. Concentrations of lactose increased and were greatest in blood collected 20h (520.4±54.1, 268.1±48.2, 345.0±52.3, 418.4±49.8 μM, for the 4 treatment groups respective to the order listed above) after supplemental ECP and final milk removal. At 40h, concentrations approached those observed 24h before dry-off (140.5±52.1, 57.6±47.1, 90.1±51.4, 61.2±48.4 μM, respectively). Concentrations of lactose at 20h were positively correlated with milk yield of cows at dry-off. Similar temporal profiles of lactose in blood plasma of cows supplemented or not with ECP indicated that ECP at dry-off did not markedly alter the course of tight junction leakage that typically occurs in mammary epithelial tissue during progressive early involution when milk removal is discontinued.

Effects of milking interval and cisternal udder evaluation in Tunisian Maghrebi dairy dromedaries (Camelus dromedarius L.)

Effects of 4 different milking intervals (8, 12, 16, and 24h) on milk yield and milk composition were studied in Tunisian Maghrebi dairy dromedaries (n=6) at late lactation [240±14 days in milk (DIM), 5.84±1.62 L/d]. Camel-cows suckled their calves for 2 mo, were hand milked while suckling until mo 4 of lactation (calf weaning) and machine milked thereafter. Intravenous injection of oxytocin was administered before machine milking at each experimental milking to induce complete milk ejection and to avoid carryover effects of milking intervals. Cisternal and alveolar milk were measured at 380±16 DIM for a 24-h milking interval. Milk accumulated logarithmically (R2=0.95) in the udder from 8- to 24-h milking interval without reaching a plateau. Consequently, milk secretion rate decreased exponentially (R2=0.93) according to milking interval. Compared with 12-h milking interval (6.1 L/d), estimated daily milk yield was 113, 87, and 70% for 8-, 16-, and 24-h intervals, respectively. Total milk solids, milk fat content, and milk pH decreased with increasing milking interval, showing the greatest value at 8-h intervals (14.1±0.4%, 4.6±0.5%, and 6.66±0.05, respectively) and the lowest at 24-h intervals (12.3±0.9%, 2.9±0.6%, and 6.54±0.02, respectively). Milk protein (3.9±0.1%), lactose (4.5±0.2%), ash (0.84±0.01%) and density (1.028±0.01) remained constant for all milking intervals. Milk K, Ca, and Mg contents increased as milking interval increased, but Na content did not change (0.06±0.01%, on average). Milk Na:K ratio tended to decrease from 0.35 (1:2.9) to 0.22 (1:4.5) for the extreme milking intervals. Plasma lactose concentration steadied from 8- to 16-h (67±32 μmol) but increased dramatically at 24-h intervals (338±118 μmol), indicating that mammary tight junctions became permeable after 24h of milk accumulation. Camel udders showed small cisterns (19.3% of total milk in the udder at 24h) when compared with other dairy animals; we recommend the use of prestimulation for machine milking and selection for larger udder cisterns. Alveolar milk contained more fat (5.16 vs. 1.75%; SEM, 0.39%) and protein (3.23 vs. 2.73%; SEM, 0.15%) than cisternal milk. Despite the increase of plasma lactose during tight junction leakiness, the tendency for the Na:K ratio to decrease may be indicative of a camel's specific regulatory mechanism for controlling Na and K concentrations in milk and delaying the inhibitory effect of milk stasis on milk secretion rate. In conclusion, this short-term study proved the low storage capacity of the Tunisian Maghrebi camel udder but also showed their moderate ability to adapt to extended milking intervals at late lactation.

Disrupted secretory activation of the mammary gland after antenatal glucocorticoid treatment in sheep

Antenatal glucocorticoids are administered to women at risk of preterm delivery to prevent neonatal respiratory morbidity. The effects of exogenous glucocorticoids on the development of lactation are unknown. This study investigated the effects of a single dose of antenatal glucocorticoids on secretory activation in sheep before and after parturition. Pregnant ewes (N=36) were randomised to receive either medroxyprogesterone acetate (MPA) at 118 days of pregnancy and betamethasone at 125 days (BETA group), MPA at 118 days and saline at 125 days (MPA group) or saline at 118 and 125 days (SALINE group). The concentration of lactose, progesterone, cortisol and prolactin in maternal plasma was measured during pregnancy. After term parturition, the concentration of lactose in milk and maternal plasma was measured daily for 5 days. Lambs were weighed at birth and at 5 days of age; milk volume was measured on day 5. The concentration of lactose in maternal plasma increased significantly after betamethasone administration, corresponding to a fall in plasma progesterone. No changes in lactose were observed in MPA or SALINE ewes. Transient decreases in cortisol and increases in prolactin were observed in the BETA group, but not in either the MPA or SALINE group. After parturition, BETA ewes experienced reduced milk yield and lamb weight gain, and delayed increases in milk lactose levels compared with MPA and saline controls. This study demonstrated that, in sheep, antenatal glucocorticoid administration disrupted secretory activation, causing precocious mammary secretion before parturition and compromising postpartum milk production and lamb growth.

Mammary gland involution and endocrine status in sows: Effects of weaning age and lactation heat stress

The effects of lactation length and/or heat stress on endocrine status of sows and indicators of mammary involution at weaning were studied. Twenty-eight first parity Yorkshire × Landrace sows were housed at 21 (CTL) or 29°C (HS) throughout lactation. Within each temperature group, half the sows were randomly assigned to a weaning age of 22 (W22: CTL, n = 7; HS, n = 8) or 44 d (W44: CTL, n = 6; HS, n = 7). Litter size was standardized to 10 or 11 on day 2 and to 9 on day 23 of lactation. Blood and milk samples were collected on days -1, 1, 2 and 3 relative to weaning (day 0). Concentrations of prolactin, IGF-I and lactose were measured in both fluids and concentrations of Na, K and IGFBP-5 were measured in milk. Standard milk composition was also determined. Lactose was lower (P &lt; 0.001) and protein greater (P &lt; 0.05) in all milk samples from W44 compared with W22 sows. Percentages of milk fat and dry matter were lower (P &lt; 0.05) in W44 than in W22 on days 2 and 3 post-weaning. Milk lactose was reduced (P &lt; 0.05) with heat stress on days 2 and 3. The day before weaning, concentrations of prolactin in blood (P &lt; 0.001) and lactoserum (P &lt; 0.01) were lower and those of IGF-I in lactoserum were greater (P = 0.01) in W44 than in W22. Values of IGFBP-5 in milk were drastically lower (P &lt; 0.001, more than a fourfold difference), those of Na greater (P &lt; 0.001) and those of K unchanged (P &gt; 0.1) for W44 compared with W22. Post-weaning, evidence of leaky tight junctions (increased milk Na, decreased K) was evident a day earlier (2 vs. 3) in W44 compared with W22 and was most marked in HS. In support, plasma lactose concentrations increased transiently on the same day in HS W44. The greater Na/K ratio in W44 compared with W22 taken together with increased plasma lactose particularly in HS shows that mammary epithelial cell tight junctions become leaky as lactation advances. Weaning age therefore has an impact on milk variables which are indicative of the status of mammary involution in sows and some of these effects are exacerbated by heat stress. Key words: Heat stress, lactation length, mammary involution, sows, weaning

Effects of endophyte infection of perennial ryegrass on somatic cell counts, mammary inflammation, and milk protein composition in grazing dairy cattle

Concentrations of selected milk and blood components were determined for dairy cows grazing perennial ryegrass pasture with either high or low levels of endophyte (Neotyphodium lolii) infection during the summers of 1998 and 1999. Cows (Jerseys in 1998 and Friesians in 1999) grazing high endophyte pasture did not exhibit any increase in somatic cell count in either study, as measured in the bulk milk of cows and in milk from individual udder quarters. Measurements of Na, K, serum albumin, and IgGj in milk, together with concentrations of lactose in blood plasma, provided no evidence that high endophyte pasture was associated with mammary inflammation. The level of endophyte infection in pasture did not alter milk concentrations of a‐lactalbumin and P‐lacto‐globulin. Similarly, no effect of endophyte on plasma prolactin concentrations was observed.

Effect of milking frequency and pasture intake on milk yield and composition of late lactation cows.

Twenty-four monozygous twinsets in late lactation (> 210 d in milk) were used to examine the effects of feed restriction and milking frequency prior to drying off on milk yield and composition in a pastoral dairying system. Cows were assigned to one of four treatment groups for 26 d and were milked either twice or once daily and given either unrestricted or restricted access to feed. Dry matter intakes averaged 16 or 8 kg per cow per day, and diets comprised ryegrass and white clover pasture supplemented with 15% pasture silage. Feed restriction and once daily milking reduced milk yield and increased concentrations of milk fat and protein. Somatic cell count was increased by feed restriction only. Production losses caused by feed restriction were nearly threefold higher than were those for once daily milking. Yields of components that were mammary synthesized and serum derived were reduced by feed restriction, in accordance with milk volume reduction. Plasma lactose concentration increased with once daily milking only and indicated enhanced permeability of mammary tight junctions. Both feed restriction and once daily milking compromised milk quality, but increased leakage of serum components into milk via mammary tight junctions was deemed to occur only for once daily milking.

Normal plasma lactose concentrations and kinetics of intravenously infused lactose in cattle

Plasma lactose concentration and its kinetics were determined in apparently normal cattle, as a prelude to investigating its chemotherapeutic significance in bovine trypanosomiasis. It is hoped that intravenously administered lactose may be able to reduce the rate of sequestration of desialylated erythrocytes during Trypanosoma vivax infection of cattle; thus decreasing the rate of development of trypanosomal anaemia in infected animals. A range of 0·061 to 0·55 mM with a mean of 0·208 ± 0·128 mM standard deviation (SD), observed in adult cattle was significantly lower (P<0·001) than corresponding values in recently weaned calves; 0·429 to 1·496 mM (0·972 ± 0·318 mM). Semi-logarithmic plots from calves given a single dose (0·5 g lactose per kg bodyweight as a solution in normal saline, infused at the rate of 18 ml min −1) showed a biexponential pattern of regression lines. Decrease in plasma concentrations was biphasic and lactose was rapidly distributed into the extravascular space after administration. The biological half-life ( t 1 2 ) of the infused lactose ranged from 4·10 to 6·00 hours (5·01 ± 0·81 hours); its mean elimination rate constant was 0·14 ± 0·02 hour −1, mean apparent volume of distribution was 168·09 ± 56·65 ml kg −1 while its mean total clearance was 23·54 ± 8·31 ml kg −1 hours. A single dose rapidly reached a peak and gradually fell below the pre-infusion level while repeated doses did not cause accumulation of the lactose in the plasma as each infusion fell back to normal relatively rapidly.

Effect of Atropine on Milk Protein Yield by Dairy Cows with Different β-Lactoglobulin Phenotypes

The objective of this study was to determine the response of individual milk proteins to a reduction in amino acid (AA) availability induced by atropine and to determine whether the response was different in cows with different β-lactoglobulin (LG) phenotypes. Six cows that were homozygous for the A variant of β-LG and six cows that were homozygous for the B variant of β-LG were each given a single subcutaneous injection of saline or 20mg of atropine. In both groups of cows, atropine decreased milk yield by 30% and reduced the concentration of α-lactalbumin (LA) by 25 to 30% at 8h following injection. Eight hours after atropine injection, yield of β-LG was 41% lower than it was following saline injection, and yield of β-casein (CN) after atropine injection declined 16% relative to saline. Concentrations of BSA and the ratio of γ-CN to β-CN, which reflects plasmin activity in milk, were significantly increased after administration of atropine. Although the response to atropine tended to be more pronounced in cows that were homozygous for β-LG B, they were not significantly different from the response of cows that were homozygous for β-LG A. The differential response of individual proteins to a reduction in AA concentrations in whole blood suggested that susceptibility to restriction in substrate availability differed for individual proteins. The concentration of lactose in plasma did not change, which implied that the integrity of the mammary epithelial barrier was not compromised when AA derived from blood were diminished. The consistent concentration of lactose combined with the minimal increase in total yield of BSA in milk following atropine treatment indicated that the increased concentration in milk of proteins derived from serum was due to the concentrating effect of lower milk volume.

Time course of milk accumulation-induced opening of mammary tight junctions, and blood clearance of milk components.

Eight cows in early lactation were used to study the effect of milk accumulation on the state of mammary tight junctions and to examine alpha-lactalbumin as an indicator of tight junction permeability in vivo. During three successive periods, the cows were milked twice (4 days), once (6 days), and twice daily (4 days). Plasma lactose, alpha-lactalbumin, and milk sodium concentrations were used as indicators of tight junction permeability. Furthermore, four cows were used to study the clearance of lactose and alpha-lactalbumin from the blood. Milk yield during once-daily milking decreased by 15.4% (P < 0.001). All indicators of mammary tight junction patency increased (P < 0.05) transiently during once-daily milking and indicated that tight junctions opened after approximately 18 h. Plasma alpha-lactalbumin and lactose were highly correlated (r = 0.82, P < 0.001), indicating the suitability of plasma alpha-lactalbumin as an indicator of tight junction status in vivo. Clearance of alpha-lactalbumin and lactose from the blood was best described by a biexponential model. Elimination half-lives for lactose and alpha-lactalbumin were 44 and 40 min, respectively. This study showed that milk stasis during early established lactation induces tight junctions to switch to a leaky state after approximately 18 h and to revert to the closed state shortly after milking.

The effect of exogenous prolactin on lactation performance of first-litter sows given protein-deficient diets during the first pregnancy

Twenty-four sows were used to study the effects of dietary protein restriction during pregnancy and exogenous porcine prolactin (pPRL) during late pregnancy and throughout lactation on lactation performance. Eight sows were given a protein-adequate diet containing 179 g crude protein (CP)kg−1 during their first pregnancy while the remaining 16 sows received the same amount of a diet containing 80 g CP kg−1. Eight of the sows given 80 g CP kg−1 during pregnancy were injected with 15 mg pPRL i.m. twice daily at 08:00 and 20:00 between day (d) 102.1 (±0.3) of pregnancy and weaning after their first lactation. Pregnant sows offered the low protein diet gained significantly less body weight during gestation and tended to eat less in the subsequent lactation than sows given the protein-adequate diet. Dietary protein had no significant effect on birth weight, milk yield, milk composition or growth rate of the litter during lactation. Neither dietary protein intake during pregnancy nor exogenous prolactin affected the concentrations of plasma glucose, serum insulin, urea or non-esterified fatty acid (NEFA) during lactation. The concentration of lactose in plasma during lactation was unaffected by treatment, but at d 105 of pregnancy, plasma lactose levels were greater in sows which had received exogenous prolactin (32.4 vs. 6.2 mg l−1, P < 0.05). The concentrations of RNA and DNA in mammary tissue biopsies were unaffected by either dietary protein or pPRL. The concentration of RNA and DNA increased between d 70 and 90 from 0.66 to 2.77 mg g−1 and from 0.54 to 1.19 mg g−1, respectively. Thereafter, RNA increased to 4.40 mg g−1 at d 14 of lactation whilst DNA concentration remained at a similar level of 0.90 mg g−1. Milk yield of sows between d 5 and 8 and between d 19 and 22 of lactation was reduced from 8.36 to 7.00 kg day−1 and from 10.74 to 8.22 kg day−1, respectively, in sows given pPRL. The protein content of colostrum from sows treated with pPRL was reduced from 164 to 104 g kg−1 whereas the fat content increased from 47 to 127 g kg−1. These results indicate that the administration of exogenous pPRL during late pregnancy and throughout lactation initiated lactogenesis prematurely and reduced subsequent milk yield during established lactation.