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Ample research has described the assessment of dimensional changes for different teat traits, whereas diagnostic techniques to reliably assess blood circulation in teats of dairy cows are limited. Here, we describe the development and evaluation of a scanning technique to quantify blood flow in teats of dairy cows using power Doppler ultrasonography. In 2 consecutive trials, 384 teat scans [trial 1, n = 256 (sagittal plane, n = 128; transverse plane, n = 128); trial 2, n = 128 (transverse plane)] from 16 cows were obtained by the same 2 operators. Perfusion intensity from single images (trial 1) and video images (trial 2) were assessed using a commercially available software program. Intraclass correlation coefficients (ICC) and concordance correlation coefficients (CCC) were used to assess interoperator reproducibility (agreement between measurements performed by different operators) and intraoperator repeatability (agreement between measurements performed by the same operator). In trial 1, interoperator ICC and CCC indicated poor agreement (ICC ≤0.26, CCC ≤0.26). Intraoperator ICC and CCC demonstrated poor agreement between duplicate measurements within operators (ICC ≤0.19, CCC ≤0.19). Modifications after trial 1 included (1) a different ultrasound device, (2) analysis of video clips rather than single images, (3) restriction to 1 sectional plane (i.e., transverse), and (4) a scanning sequence such that repeated scans within operators were measured one after another. Through these modifications, intraoperator repeatability in trial 2 yielded fair to good agreement, with intraoperator ICC and CCC over both operators ranging from 0.44 to 0.70 and from 0.57 to 0.69, respectively, whereas interoperator ICC and CCC showed poor agreement (ICC = 0.35, CCC = 0.34). We conclude that repeatable measurements of blood perfusion intensity of teats in dairy cows can be attained with power Doppler ultrasonography. Power Doppler ultrasonography is a suitable tool to quantify slow flow in small vessels and may be an acceptable diagnostic technique to assess changes in blood circulation that result from machine milking in teats of dairy cows, although further research is necessary to validate this hypothesis., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Objectives were to investigate the effects of supplementation with corn dried distiller's grains plus solubles (DDGS) to late gestating beef cows on arterial blood flow to the mammary glands during late gestation and early lactation; colostrum and milk production; dystocia and immunity; and calf BW. Cows were fed a control (CON; n = 15; 5.1% CP; 36.2% ADF) diet consisting of 90% corn stover and 10% corn silage on a dry basis offered ad libitum or CON diet with supplementation of DDGS (0.30% of BW; SUP n = 12). Mammary gland blood flow was assessed on day 245 of gestation. At parturition, maternal and calving parameters were assessed; colostrum and jugular blood was sampled; and dams were weighed. Mammary gland blood flow and milk production was measured on day 44 of lactation. Calves were weighed fortnightly for 8 wk and at weaning. Colostrum production tended to be greater in SUP dams than in CON dams (837 vs. 614 ± 95 g, P = 0.10). Calves of SUP dams were heavier at birth and 24 h (0 h, 43.2 vs. 39.8 ± 1.0 kg, P = 0.02; 24 h, 44.0 vs. 40.4 ± 1.1 kg, P = 0.02). At birth and 24 h, blood pCO2 was greater in calves born to SUP dams (6.82 vs. 6.00 ± 0.41 kPa, P = 0.04). Serum IgG did not differ (P = 0.21) at 24 h. Ipsilateral mammary gland blood flow of SUP cows was greater than CON cows (2.76 vs. 1.76 ± 0.30 L/min; P = 0.03); however, when summed with contralateral, total blood flow was similar (P = 0.33). Hemodynamic measures on day 44 of lactation were similar (P ≥ 0.32). Milk production tended to be increased (13.5 vs. 10.2 ± 1.2 kg/d, P = 0.07) in SUP vs. CON cows. Despite similar BW through 56 d, calves from SUP cows were heavier (P = 0.04) at weaning (309.7 vs. 292.0 ± 6.0 kg). In conclusion, we accept our hypothesis that DDGS supplementation during gestation influenced mammary blood flow, milk production and calf weights. These findings implicate maternal nutrition's leverage on both nutrient and passive immunity delivery to the calf early in life as well as potential advantages on long-term performance., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

During lactation, mammary epithelial cells (MECs) form the blood-milk barrier by less-permeable tight junctions (TJs) to prevent the leakage of milk components. Phytoestrogens affect the proliferation, differentiation, and apoptosis of MECs. However, it remains unclear whether phytoestrogens are involved in the blood-milk barrier. Therefore, we investigated the influence of phytoestrogens (coumestrol, genistein, and daidzein) by using an in vitro mouse-MEC-culture model. The results showed that coumestrol and genistein changed the expression of TJ proteins (claudins-3 and -4 and occludin), weakened barrier function, and reduced β-casein production. Daidzein also weakened barrier function without inhibiting β-casein production. Additionally, coumestrol and genistein induced apoptosis in MECs. These results indicate that phytoestrogens weaken the blood-milk barrier by directly affecting TJs and the cellular viability of lactating MECs in different ways.

The objective of this study was to clarify the effect of different milking frequencies under an automatic milking system (AMS) on milk yield, plasma metabolite profiles and mammary arterial-venous (A-V) differences of milk precursors by mammary tissues in early lactation cows. Twelve Holstein cows were divided into two and four times milking frequency treatments by AMS after calving to 50 days postpartum. Cows were given a partial mixed ration ad libitum and a concentrate diet at every milking. Dry matter intake increased similarly in both treatments with advancing postpartum days. Milk yield was greater (P < 0.001) by 25% with four times milking, but milk composition was not affected by milking frequency. Body weight change was also not affected by milking frequency. Arterial concentrations of glucose and glutamate were lower (P < 0.05) for four times milking frequency. However, arterial concentration of nonesterified fatty acids did not differ between treatments. Although mammary A-V differences of plasma concentration for most milk precursors did not differ between treatments, estimated plasma flow was higher (P < 0.05) for four times milking frequency. These results indicate that higher milking frequency may increase mammary uptake of milk precursors, whereas may not affect the extent of fat mobilization of early lactating cows from day 20 postpartum onward., (© 2014 Japanese Society of Animal Science.)

Low concentrations of the essential amino acid histidine in circulation have been shown to increase mammary blood flow and it has been suggested that this effect is mediated by histamine. The hypotheses tested in this experiment were that interstitial histamine concentrations in the mammary gland are related to arterial His concentrations and that mammary blood flow is reduced by extracellular histamine via H(1) receptors. The hypotheses were tested by infusing saline or chlorpheniramine, a blocker of the H(1) histamine receptor, into the arterial supply of the mammary glands of lactating cows infused with 44 g/h of amino acid mixtures with or without His for 10 h. Infusates were administered in a 2 x 2 factorial arrangement within a 4 x 4 Latin square to 4 multiparous Holstein cows in mid lactation. Exclusion of His from the infusate decreased protein content in milk from the infused udder half from 3.98 to 3.77%, and increased arterial alpha-aminonitrogen concentration from 3.2 to 3.4 mM. Neither the decreased arterial His concentration nor the H(1) blocker affected plasma flow to the infused udder half. We conclude that histamine is not involved in the regulation of mammary blood flow. The H(1) blocker decreased milk production in the infused udder half from 4.6 to 3.5 kg without affecting protein, fat, and lactose percentages, suggesting an inhibition of milk ejection. Cows on chlorpheniramine ate less feed during the infusion than saline-infused cows, which resulted in lower arterial concentrations and mammary uptakes of acetate. The efficiency of plasma triacylglycerol uptake across the mammary glands was decreased by chlorpheniramine but net uptake of long-chain fatty acids was not affected. The mechanism by which an amino acid deficiency influences mammary blood flow does not involve histamine signaling through the H(1) receptor and remains unidentified.

Reports over the past decade have indicated that normal lactational performance can be achieved in genetically superior and high-producing dairy cows, even when the dry period between 2 lactations is omitted. The hypothesis tested in this experiment was that normal lactogenesis I and metabolic function may be achievable in continuously milked high-yielding dairy cows as a result of the genetic selection for lactation performance and hence longevity of mammary epithelial cells. The milk production and mammary nutrient uptake in response to omission of the dry period for cows with an expected peak milk yield higher than 45 kg/d were studied in 28 Holstein dairy cows managed without bovine somatotropin. Performance and metabolic parameters were followed in late gestation and in the following early lactation. Fourteen cows were milked continuously throughout late gestation, and another 14 dairy cows experienced a 7-wk prepartum dry period. Continuous milking during the prepartum period reduced milk production in the following early lactation period by >20%. The reduced milk production could not be readily ascribed to inefficiency of the mechanisms responsible for nutrient uptake by the lactating mammary epithelial cells, nor to systemic endocrine changes. This suggests that lowered mammary nutrient uptake must have been associated with reduced mammary blood flow, metabolic activity, or both, most likely as a result of disturbed lactogenesis I prepartum or lactogenesis II postpartum triggered by as yet unknown local mechanisms. Milk protein content was elevated by 0.4 percentage units in the continuously milked cows. The underlying reason is unknown, but given the current pricing system for milk, it deserves to be further investigated.

The aim of this study was to gain a clearer understanding of the different levels of regulation involved in the reduction in milk yield in response to once-daily milking and feed restriction. The treatments were designed as a 2 x 2 factorial arrangement of 2 milking frequencies (once- or twice-daily milking) and 2 feeding levels (70 or 98% of requirements determined 1 wk before the trial). The cows were surgically prepared to study the net mammary balance of the nutrients that are precursors of milk components. Mammary efficiency in synthesizing milk components was estimated using a milk output:mammary uptake ratio. No interaction was observed between the effects of milking frequency and feeding level on milk and blood parameters except for milk protein yield, milk fatty acid profile, and nonesterified fatty acids metabolism. Once-daily milking and feed restriction reduced milk yield by 5.1 and 2.9 kg/d and fat-corrected milk yield by 4.2 and 4.1 kg/d, respectively. Both treatments induced a decrease in mammary blood flow. Once-daily milking led to a reduction in the extraction rate of glucose but no changes to the lactose output:glucose uptake ratio. Feed restriction did not change the glucose extraction rate but tended to improve the lactose output:glucose uptake ratio. Under once-daily milking, the slight increase in milk fat content (0.34 percentage units) was linked to a depressed uptake of glucose and acetate but without any variations in the uptake of beta-hydroxybutyrate and total glycerol and in the efficiency of acetate and beta-hydroxybutyrate conversion to short- and medium-chain fatty acids in milk. The decline in milk fat and protein contents (-0.43 and -0.23 percentage units, respectively) under feed restriction was associated with relatively similar reductions in the mammary uptake of all nutrients and with enhanced conversion of the glucose taken up by the mammary gland and used for lactose synthesis. As a result, once-daily milking and feed restriction seem to affect milk yield through mechanisms that may be different and relatively independent.

Recent surveys have identified the presence of perchlorate, a natural compound and environmental contaminant, in forages and dairy milk. The ingestion of perchlorate is of concern because of its ability to competitively inhibit iodide uptake by the thyroid and to impair synthesis of thyroid hormones. A recent study established that milk perchlorate concentrations in cattle highly correlate with perchlorate intake. However, there is evidence that up to 80% of dietary perchlorate is metabolized in clinically healthy cows, thereby restricting the available transfer of ingested perchlorate into milk. The influence of mastitis on milk perchlorate levels, where there is an increase in mammary vascular permeability and an influx of blood-derived components into milk, remains unknown. The present study examined the effect of experimentally induced mastitis on milk perchlorate levels in cows receiving normal and perchlorate-supplemented diets. Over a 12-d period, cows were ruminally infused with 1 L/d of water or water containing 8 mg of perchlorate. Five days after the initiation of ruminal infusions, experimental mastitis was induced by the intramammary infusion of 100 microg of bacterial lipopolysaccharide (LPS). Contralateral quarters infused with phosphate-buffered saline served as controls. A significant reduction in milk perchlorate concentration was observed in the LPS-challenged glands of animals ruminally infused with either water or perchlorate. In control glands, milk perchlorate concentrations remained constant throughout the study. A strong negative correlation was identified between mammary vascular permeability and milk perchlorate concentrations in LPS-infused glands. These findings, in the context of a recently published study, suggest that an active transport process is operative in the establishment of a perchlorate concentration gradient across the blood-mammary gland interface, and that increases in mammary epithelial and vascular endothelial permeability lead to a net outflow of milk perchlorate. The overall finding that mastitis results in lower milk perchlorate concentrations suggests that changes in udder health do not necessitate increased screening of milk for perchlorate.

Mammary arterious-venous differences (A-V) and excretion into milk of four prostanoids were related to changes in milk yield and milk vein blood velocity (MBV) in goats at different stages of pregnancy and lactation, and during somatotropin (ST) treatment in mid-lactation. Arterial concentrations and mammary A-V for the vasodilators prostacyclin (PGI(2)) and prostaglandin (PG) E(2) (measured as 6-keto-PGF(1 alpha) and bicyclic PGE(2), respectively) decreased from late pregnancy to lactation. A-V were negatively correlated to MBV (r = -0.32 to -0.34). Arterial concentrations of the vasoconstrictors PGF(2 alpha) and TXA(2) (measured as TXB(2)) changed similarly, but no A-V across the mammary gland were found. The vasodilator to vasoconstrictor ratio in plasma was around 1:1, and in skimmed milk around 0.29-0.49 due to significantly higher TXB(2) levels in milk compared to plasma. Close linear correlations were established between milk yield and excretion of TXB(2) into milk (r = 0.80, P < 0.001), and between MBV and PGE(2) excretion into milk (r = 0.69, P < 0.001). ST treatment stimulated MBV and mammary prostanoid supply, and decreased prostanoid concentration in milk vein plasma. The high arterial levels of prostaglandins during pregnancy most likely reflected uterine synthesis. Our results support a role for PGI(2) and PGE(2) in local mammary blood flow regulation during lactation. Increased mammary uptake of these two prostanoids may be involved in the mammary blood flow response to ST. TXA(2) may be synthesized by mammary epithelial as well as vascular cells, and TXA(2) may be an important factor in regulation of mammary function.

To define dietary indispensable amino acid (IAA) needs for milk synthesis by the mammary glands (MG), 16 lactating sows were fed 1 of 4 isocaloric diets varying in protein concentrations (from 78 to 235 g/kg) with an ideal amino acid (AA) pattern. On d 9, 13, 17, and 21 of lactation, blood samples were obtained simultaneously from a carotid artery and the main mammary vein every 30 min over 6 h. A quadratic regression model of the log mammary arteriovenous difference (AVD) of plasma IAA (ŷ) against daily intake of dietary IAA (X) was established. First, the reverse log intercept, defined as the mammary AVD at zero dietary AA supply, was used to quantify the contribution of endogenous IAA. The quantification was validated by body N balance coupled with AA composition analysis. Then, the estimated vertex (ŷ(max), X(i)) was used in 2 aspects: 1) The maximal mammary uptake of plasma IAA, quantified by multiplying the maximal mammary AVD and plasma flow rate, was considered the physiological IAA need for milk synthesis. 2) Corresponding to the ŷ(max), dietary IAA intake (X(i)) would represent the total dietary IAA requirement, i.e., the sum of maintenance need and milk synthesis need after adjustment for body weight loss. Thus, dietary IAA needs for milk synthesis were derived. Moreover, the estimate of lysine need for milk synthesis in this study was identical to an estimate obtained from multiple regression analysis of feeding trial data. We conclude that dietary IAA needs for milk synthesis can be quantified by the maximal uptakes of plasma IAA by porcine MG.

Objectives were to measure net fluxes of free (FAA) and peptide bound amino acids (AA) (PBAA) across portal-drained viscera (PDV), liver, splanchnic, and mammary tissues, and of milk AA output of lactating Holstein cows (n = 6, 109 +/- 9 d in milk) as influenced by flaking density of corn grain. Cows were fed alfalfa-based total mixed ration (TMR) containing 40% steam-flaked (SFC) or steam-rolled corn (SRC) grain. The TMR were offered at 12-h intervals in a crossover design. Six sets of blood samples were obtained from indwelling catheters in portal, hepatic, and mammary veins and mesenteric or costoabdominal arteries every 2 h from each cow and diet. Intake of dry matter (18.4 +/- 0.4 kg/d), N, and net energy for lactation were not altered by corn processing. Milk and milk crude protein yields (kg/12-h sampling) were 14.2 vs. 13.5 and 0.43 vs. 0.39 for cows fed SFC or SRC, respectively. The PDV flux of total essential FAA was greater (571.2 vs. 366.4 g/12 h, SEM 51.4) in cows fed SFC. The PDV flux of total essential PBAA was 69.3 +/- 10.8 and 51.5 +/- 13.2 g/12 h for cows fed SFC and SRC, respectively, and differed from zero, but fluxes of individual PBAA rarely differed between treatments. Liver flux of essential FAA was greater in cows fed SRC, but only the PBAA flux in cows fed SRC differed from zero. Splanchnic flux of FAA and PBAA followed the pattern of PDV flux, but variation was greater. Mammary uptake (g/12 h) of total essential FAA was greater in cows fed SFC than SRC (224.6 vs. 198.3, SEM 7.03). Mammary uptake of essential PBAA was 25.0 vs. 15.1, SEM 5.2, g/12 h for cows fed SFC or SRC, respectively, and differed from zero in half of the PBAA. Milk output of EAA was 187.8 vs 175.4, SEM 4.4 g/12 h in cows fed SFC and SRC, respectively, and output of most essential AA consistently tended to be greater in cows fed SFC. It is apparent that PBAA comprise a portion of total AA flux across PDV and are affected by grain processing. Further, this pool supplies an important component of AA taken up by the mammary gland. Quantifying the contribution of PBAA may improve diet formulation with respect to intestinal absorption and mammary uptake of AA.

Epithelial cells of mammary alveoles may he considered as a bioreactor that works being monitored by regulatory system maintaining balance between blood substrate supply and rate of macromolecular synthesis. In the trial performed on lactating cows assigned to feeding regimen with temporally altered level of nutrition, functioning of this system in the course of a transition period was studied. The earliest sign of adaptation was a decrease in volume blood flow through mammary gland provoking decline in uptake of water soluble substrates (glucose, amino N, P-hydroxybutyrate) with increase in arterio-venous difference across mammary gland and extraction efficiency. At the end of deprivation period, an activity of transport into the cell decreased for amino N from 7.5 to 4.5; l/min (p < 0.05), for P-hydroxyhutyrate from 16.0 to 13.8 l/min; activity of glucose transport was not changed. The data obtained indicate existence of defined points in the system monitoring organ blood supply and transport of substrates into the cell being adjusted during adaptation to alteration in the level of nutrition.

Two groups of four Holstein cows, one in their second and the other in their third or fourth lactation, were used to study temporal variations of mammary metabolism over a 12-h period between two milkings. Blood samples were collected every 30 min from an artery and a mammary vein during a 12-h interval between two milkings. Isoleucine, leucine, lysine, methionine, and phenylalanine mammary net fluxes varied or tended to change over time after milking with a similar pattern between whole blood and plasma. For these amino acids, whole blood and plasma net fluxes reached their maximum over the first 8 h after milking. Simultaneously, respiratory quotients decreased linearly and varied from 2.31 to 2.01 during the first 8 h of the period, suggesting active mammary lipogenesis. From 8 to 12 h after milking, mammary amino acid net fluxes decreased, while mammary oxygen uptake tended to increase with a concomitant decrease in the respiratory quotient reaching 1.84 to 1.40. These findings suggest that, beginning 8 h after milking, mammary uptake of amino acids starts to decrease and catabolic processes appear promoted; this phenomenon could help to explain the increase in milk production reported in the literature with increased milking frequency.

The effect of intestinal glucose supply on whole body rate of glucose appearance (WBGRa) and mammary utilization of glucose was studied in four lactating dairy cows. Glucose (0, 443, 963 and 2398 g/d) was continuously infused in the duodenum over 14-d periods using a Latin square design. A grass silage-based diet was formulated so that treatments were isoenergetic and isonitrogenous and contained 100 and 110% of energy and protein requirements according to INRA (1989). The WBGRa was measured by the [6,6-(2)H2]glucose dilution technique, and mammary glucose balance by arteriovenous differences and blood flow measurements. Duodenal glucose infusion increased arterial glucose concentrations linearly, whereas arterial concentrations of insulin, growth hormone, and glucagon were not changed. The WBGRa increased linearly with increasing glucose loads. The increase represented 42% of the intestinal glucose supplement. Mammary blood flow dramatically increased (up to 45%) and was associated with a significant increase of arterial insulin-like growth factor-1 concentrations. Mammary gland rate of glucose disappearance ([6,6-(2)H2]glucose measurement) increased linearly, whereas net mammary balance of glucose, lactose, and milk yields increased quadratically. Net mammary balance of glucose accounted for 60% of WBGRa, except for the greatest dose (47.6%). The decrease in milk yield with 2398 g/d of glucose may be explained by an imbalance in intracellular intermediate concentrations. The milk ratio of glucose-1-phosphate to glucose-6-phosphate decreased significantly at the greatest infusion of glucose. In conclusion, exogenous glucose supply to a grass silage-based diet increased WBGRa, mammary utilization of glucose and milk synthesis.

Concentrations of glucose in the external iliac artery feeding one udder half of 14 midlactation Holstein cows were increased by infusion to test the following three hypotheses of mammary function: 1) that mammary glands control their blood supply to maintain intracellular energy balance, 2) that milk precursors are taken out of capillary blood according to mass action kinetics, and 3) that the rate of milk component synthesis is dependent on its precursor's uptake from blood. The first seven cows received 20 g/h glucose during 10 h of infusion. Arterial concentrations of glucose were locally increased by only 10%, and the iliac plasma flow was not affected by glucose infusion, so the next seven cows were given 90 g/h glucose. Quantitative predictions resulting from the hypotheses were that arterial plasma flow would decrease by 32% with 90 g/h glucose infusion, glucose uptakes would increase and acetate, fatty acid, and amino acid uptakes decrease, and milk protein and fat yields and percentages would decrease. Iliac plasma flow decreased 16%, half of what was predicted, which suggests that other regulatory processes besides blood flow control took part in the response. Acetate and fatty acid uptakes by the mammary glands were reduced as predicted because of the lower blood flow, but an unexpected depression in extraction of plasma triacylglycerol also contributed to the reduced fatty acid uptake. Milk fat and protein yields were not affected by the exogenous glucose, falsifying the third hypothesis that milk component secretion is a function of uptake of its precursor. Milk fat and protein percentages declined with glucose infusion because of increased lactose synthesis and secretion of water into milk.

Objectives were to determine net release or uptake of alpha-amino N, ammonia N, and urea N across portal-drained viscera, liver, splanchnic, and mammary tissues of lactating Holstein cows (n = 8, 86 +/- 8 d in milk) fed alfalfa hay-based total mixed rations containing 40% dry-rolled or steam-flaked sorghum grain. The total mixed rations were offered at 12-h intervals in a crossover design. Blood samples were obtained from indwelling catheters in the portal, hepatic, and mammary veins and mesenteric or costoabdominal arteries, every 2 h for each cow and diet. Steam-flaking increased in vitro rate of starch hydrolysis compared with dry-rolled sorghum (66 vs. 25%). Diet did not alter dry matter intake (18.2 +/- 0.3 kg). Daily milk yield (27.6 +/- 0.8 kg), efficiency of production, and most milk components did not differ between diets, but fat yield was reduced (0.86 vs. 0.91 kg/d) by steam-flaked sorghum, and lactose concentration was increased (4.99 vs. 4.82%). Blood flows in portal and hepatic veins did not differ between diets. Steam-flaking tended to increase urea N cycling to the gut (162 vs. 95 g/d) compared with dry-rolling of sorghum, whereas net absorption of ammonia N and alpha-amino N across portal-drained viscera were decreased. Net mammary uptake of a-amino N increased more than 20% (83 vs. 67 g/d), resulting in a higher mammary extraction ratio (15 vs. 11%) for steam-flaked versus dry-rolled sorghum. Flaking of sorghum improved the efficiency of postabsorptive N metabolism by increasing urea N cycled to the gut and alpha-amino N uptake by the mammary gland.

Four multiparous Holstein cows (569+/-122 kg) surgically prepared with indwelling catheters in the mesenteric, portal, and hepatic veins and carotid artery were allocated in a 4 x 4 Latin square to determine the effects of dietary crude protein (CP) level and amino acid (AA) profile on N metabolism during early lactation (from 25 to 65 d in milk). Cows received their diets in two equal meals and were milked twice daily. The dietary treatments were: 18% CP with a high (18H) or a low (18L) quality AA profile, and 15% CP with a high (15H) or a low (15L) quality AA profile. The four diets were similar in net energy for lactation (1.75 NEL Mcal/kg) and contained the same amount of RUP (34% of CP). The quality of the AA profile pertained only to the essential AA (EAA), and was assessed by comparison with the EAA profile of casein and considered the potential contribution of EAA from ruminal bacteria. The 18H and 15H diets were supplemented with 50 and 25 g/d of ruminally protected Met, respectively. After 10 d on treatment, a blood flow marker (p-amino-hippurate) was infused into a mesenteric vein, and arterial, portal, hepatic, and mammary blood samples were obtained at 3, 6, and 12 h after feeding. Dry matter intake was similar across treatments (23.4+/-0.5 kg/d). Amino acid oxidation, and consequent urea production, in the liver were numerically greater with the 18% CP rations, and, as a result, arterial urea concentrations were greatest (P < 0.01) with these rations. The amount of total AA extracted by the mammary gland tended to be greater with the H than with the L diets (21.4 vs. 18.2 mmol/ h, respectively). Milk yield tended to be greater (P = 0.16) with the 18H and 15H diets (47.7 and 46.3 kg/d, respectively) compared with the 18L and 15L diets (45.9 and 44.6 kg/d, respectively). Also, milk CP and casein contents were greatest (P = 0.09) with the H diets compared with the L diets. Milk and plasma urea N were greatest (P < 0.01) with the 18% CP diets. The efficiency of N utilization for milk protein synthesis was greatest (P < 0.09) with the 15% CP diets. It is concluded that milk protein production during early lactation is less susceptible to variations in dietary CP contents than variations in the AA profile of the dietary protein.

epithelium. Neutrophil migration across mammary arterial endothelial cells was almost completely dependent on CD18, the beta-chain of the beta(2) integrins, and to a lesser extent on CD11b, one of the alpha-chains of the beta(2) integrins. Neutrophil migration across collagen was partially blocked by monoclonal antibodies to CD18. No inhibition was observed by monoclonal antibodies to CD11b. Conversely, neutrophil diapedesis across mammary epithelial cells was dependent to a greater extent on CD11b. These results provide evidence for different CD11b/CD18-dependent mechanisms for neutrophil diapedesis across the various cell layers of the blood-milk barrier.

The effect of dietary lipids on the fatty acid (FA) profile of cows' milk fat is mainly dependent on digestive processes and mammary gland uptake and metabolism of FA. The objective of this study was to determine the separate effects of high arterial concentrations of 16:0, 18:0 and cis-18:1(n-9) on uptake, synthesis and 18:0 desaturation rate in the mammary gland of lactating dairy cows, via arterio-venous differences and mammary gland balance of FA. In a 4 x 4 Latin square, four lactating Holstein cows with cannula in the proximal duodenum were infused duodenally with a mixture providing daily 0 (C treatment) or 500 g FA with mainly 16:0 (P treatment), 18:0 (S treatment) or cis-18:1(n-9) (O treatment). Significantly higher arterial concentrations of infused FA in arterial plasma nonesterified FA and triglycerides (NETGFA) were observed with P and O treatments, but the effect of the S treatment was much lower. Arterio-venous differences of NETGFA increased with arterial concentrations. The number of synthesized FA in the mammary gland was not significantly affected by duodenal infusion of FA. Mean chain length was significantly reduced by P and O treatments, suggesting an effect of mammary gland uptake of long-chain FA on the termination process of mammary gland synthesis of FA. Across all treatments, 4:0 mammary gland balance increased linearly (r = 0.67, P = 0.004) with mammary gland FA uptake. Mammary gland desaturation of 18:0 to cis-18:1(n-9) averaged 52% and was not significantly affected by treatments, but was reduced by trans-18:1 mammary gland uptake. Uptake, synthesis and desaturation of FA by the mammary gland of dairy cows are affected by arterial concentrations of 16:0, 18:0 and cis-18:1(n-9).

Ten Holstein cows in early lactation were used in a replicated 5 x 5 Latin square design to study the effects of MgO and three buffers added to diets containing Ca salts of canola oil fatty acids. Treatments were 1) control (basal diet; no buffer). 2) 1.1% NaHCO3 plus 1.1% KHCO3, 3) 1.9% NaHCO3, 4) 0.5% MgO, and 5) 2.0% Na sesquicarbonate (percentage of dry matter). The control diet contained 53% grass silage, 43% concentrate, and 4% Ca salts. Body weight, intake, milk yield, and percentages of milk fat, protein, and lactose were unaffected by treatments. Buffers and MgO tended to increase triacylglycerol extraction by the mammary gland and changed the proportions of some fatty acids in milk. Arterial concentrations of acetate and triacylglycerol were correlated with their respective arteriovenous differences. Extraction by the mammary gland was high for acetate (approximately equal to 58.2%), triacylglycerol (approximately equal to 47.3%) propionate (approximately equal to 34.6%), and glucose (approximately equal to 24.3%). Extraction of free fatty acids, phospholipids, or cholesterol was negligible. Mammary triacylglycerol arteriovenous difference tended to be higher than when MgO was fed than when NaHCO3 was fed. Sodium sesquicarbonate, NaHCO3, and the blend of bicarbonate buffers increased C18:2 in milk fat when compared with the control treatment. The concentration of C18:2 in milk fat decreased when MgO was fed, but the ratio of cis-C18:1 to trans-C18:1 increased compared with effects of dietary NaHCO3. Medium-chain fatty acids in milk fat tended to be higher than Na sesquicarbonate than with NaHCO3. Buffers and MgO modified the profiles of fatty acids in milk.

Human synthetic parathyroid hormone-related protein (PTHrP) increased mammary blood flow (MBF) following close-arterial infusion via the external pudic artery in goats during mid-lactation. MBF increased 74 +/- 8% within 30 min of the start of continuous infusion of PTHrP compared with 10 +/- 3% in controls. MBF decreased by 90 min, however, and was not different from control values for the remainder of the infusion. The increase in plasma concentrations of calcium and decrease in phosphate during PTHrP suggests that this was not due to altered activity of PTHrP, but may relate to downregulation of response or production of counter-regulatory vasoconstrictive agents within the gland. This problem was alleviated when PTHrP was infused in a pulsatile fashion. An average 14-40% increase in MBF was achieved over 6 h, but this did not alter the rate of milk secretion, suggesting that mammary hyperaemia is not sufficient by itself to increase milk yield in the normally lactating goat. MBF increased in a dose-dependent fashion, although the lowest dose used to give a detectable response was approximately 40-fold higher than the concentration normally present in the mammary venous circulation. Thus, endogenous PTHrP may not be an important regulator of MBF during lactation in the goat.

1. Glucose and protein uptake were measured in both mammary glands of two low- and two high-yielding dairy goats during lactation. 2. Low-yielding goats tended to have higher arterial glucose concentrations, but approximately 40% lower arterio-venous differences (AV) and extraction rates (E) for glucose than high-yielding goats. 3. AV and E for glucose (but not protein) were linearly related to yields of both lactose, milk protein and fat. 4. Mammary uptake of glucose is determined primarily by mammary glucose metabolism, not glucose supply; lower intracellular glucose concentration in mammary cells of genetically superior animals thus explains the more efficient mammary uptake.

The milk yield and mammary blood flow responses to close-arterial, intra-mammary infusion of IGF-I were investigated in five Saanen goats milked frequently or normally the day before. Animals were infused for 6 h with recombinant human IGF-I (1.3 nmol/min) and milked hourly following i.v. injection of oxytocin beginning 2 h before infusion and then every 2 h. On one occasion animals were milked five times (after i.v. injection of oxytocin) on the day before infusion and on the other they were milked twice, without oxytocin. The ratio of milk yield from the infused to that from non-infused gland increased by 17 +/- 4% (mean +/- S.E.M.) in goats milked twice the day before infusion and by 6 +/- 2% when the infusion was preceded by frequent milking. Maximal responses were obtained 4 h after the start of the infusion and differed significantly (P < 0.05), according to pretreatment milking. Blood flow through the infused gland rose in parallel to the milk yield response. At 5 h, when maximal levels were achieved, blood flow was 182 +/- 23% of the pre-infusion flow rate following twice-daily milking and 139 +/- 3% of the pre-infusion flow rate following more frequent milk removal. Thus, more frequent milk removal on the day before close-arterial infusion of IGF-I attenuated both the milk yield and mammary blood-flow response to the infusion of IGF-I.

Milk yield is determined by number, size, synthetic and secretory capacity of mammary alveolar cells and low amounts of residual milk after milking. Initial milk yield and persistency of lactation are important. Important factors to achieve high milk yield are: optimal mammary development and preparation to lactation; maintenance and possibly increase in number and of synthetic capacity of alveolar cells during established lactation; sufficient availability of substrates for milk synthesis through high feed intake, readiness to mobilize body reserves (especially depot fat) and increased mammary blood flow; optimal milk let-down; regular and possibly more than twice milking/day. Favorable for a high milk yield are high circulating concentrations of somatotropin, possibly associated with increased production and levels of the insulin-like growth factor I, in the presence of low concentrations of and reduced sensitivity or responsiveness of target organs to insulin, reduced circulating levels of thyroid hormones and possibly enhanced conversion of thyroxine to triiodothyronine in the mammary gland. Enhanced sensitivity of depot fat to lipolytic hormones is favorable for high milk production.

125I-Labelled insulin-like growth factor-I (IGF-I) was infused as the free form directly into the pudic artery supplying one gland of lactating goats (n = 6). The infusion was for 60 min and 0.4 +/- 0.09% (S.E.M.) of the infusate was secreted into milk from the infused gland during its first passage through that gland. A large proportion of the 125I-labelled IGF-I escaped into the systematic circulation and was secreted into milk of both glands. A total of 5.2 +/- 0.4% of infused radioactivity was recovered in milk from both glands from 0 to 720 min. Radioactivity consisted of trichloroacetic acid (TCA)-precipitable and -soluble counts which were shown by gel filtration to be authentic IGF-I and degraded products of the peptide. The amount and time course of TCA-soluble radioactivity in milk from both glands was similar, suggesting degradation of 125I-labelled IGF-I at extramammary sites. Maximum specific activity for 125I-labelled IGF-I in milk from the infused gland was reached 80-120 min after the start of infusion and was 2.5-fold greater than milk from the non-infused gland. The time course of appearance of 125I-labelled IGF-I in milk suggests that transfer was via the transcellular pathway and this was further supported by comparing the pattern of transfer of [14C]sucrose and [14C]amino acids. When excess unlabelled IGF-I was included in the infusate, specific activity in milk from the infused gland was reduced to that of the non-infused gland, indicating a competitive and saturable mechanism of secretion for 125I-labelled IGF-I.(ABSTRACT TRUNCATED AT 250 WORDS)

The close-arterial infusion of free insulin-like growth factor-I (IGF-I; 1.1 nmol/min) for 6 h into the pudic artery supplying one mammary gland of lactating goats caused a 25 +/- 6% (mean +/- S.E.M., n = 6) increase in the rate of milk secretion of that gland. The increase in the rate of milk secretion in the adjacent noninfused gland (14 +/- 4%) was not significantly different from that observed during saline infusion (4 +/- 5%). Blood flow to the infused gland was increased from 378 +/- 26 ml/min 1 h before to 487 +/- 56 ml/min approximately 5 h after the start of the infusion of IGF-I, declining to 420 +/- 44 ml/min approximately 2 h after the end of the infusion. The total concentration of IGF-I (free and bound) in milk of the infused gland was significantly higher than that of the non-infused gland. The concentrations of IGF-I in carotid arterial plasma samples increased during IGF-I infusion from a mean value of 32 +/- 2 nmol/l before to a maximum of 49 +/- 3 nmol/l 5 h after the infusion commenced. Circulating concentrations of total IGF-I declined slowly after the infusion with an estimated half-life of 5 h. Infusion of saline alone did not alter mammary blood flow or the concentration of total IGF-I in milk or plasma. The results indicate that the infusion of free IGF-I into the mammary arterial supply enhances milk secretion and mammary blood flow in intact, conscious goats.(ABSTRACT TRUNCATED AT 250 WORDS)

1. Measurements of milk secretion, mammary oxygen consumption and blood flow, cardiac output and blood pressure have been made in lactating goats exposed to a thermoneutral environment and to two levels of cold stress which were sufficient to raise total body oxygen consumption by an average of 18 and 46% above that in a thermoneutral environment. 2. Mammary blood flow and oxygen consumption were not appreciably affected by the milder cold stress but fell to 63 and 70%, respectively, of their thermoneutral value in the moderately cold environment. Resistance to blood flow through the udder was increased by this level of cold stress and the gland took a smaller fraction of an increased cardiac output. 3. Both levels of cold exposure caused a reduction in milk secretion, to 80 and 40% of its thermoneutral value in the mild and moderate cold respectively. The amount of milk secreted per unit volume of blood flowing through the gland decreased as the severity of the cold stress increased. 4. Voluntary food intake was not affected by the change in environmental temperature but the intake of water was reduced to 55% of the thermoneutral value during mild cold exposure and to 40% of the thermoneutral value during the moderate cold stress. 5. It is concluded that a reduction in blood flow to the mammary gland could be one of several possible mechanisms underlying the depression of milk secretion during cold exposure.

1. The effects of raising intramammary pressure on mammary function have been studied in conscious goats. 2. When intramammary pressure was raised to levels which normally occur following cessation of milking (by infusing isosmotic sucrose into the lumen of the gland) the rate of milk secretion fell within 6 hr, but in short-term studies at these pressures, there was no reduction in mammary blood flow (in fact there was a significant increase) and no change in oxygen consumption or glucose uptake. 3. At pressures more than twofold higher than those which occur under physiological conditions, there was a decrease in mammary blood flow within 5 min; in addition the arterio-venous difference for oxygen, but not for glucose, fell. The changes were reversed when pressure was lowered. Similar results were obtained in autotransplanted (denervated) glands. 4. Intramammary pressure-volume curves were determined 3 days before cessation of milking. By determining the rate of secretion after cessation it was evident that the rate of secretion fell as the pressure-volume curve steepened. 5. A positive correlation was found between empty gland volume and functional capacity of the gland (volume of liquid needed to raise intramammary pressure to 30 mmHg at the tip of the teat) in goats of the Saanen breed. 6. No apparent effect on blood-milk potential difference was obtained at pressures within the range observed after cessation of milking. At higher pressures, the potential difference fell, eventually to zero. 7. It is concluded that in the goat the arrest of milk secretion following cessation of milking is caused by mammary distension but is not due to a primary effect of mammary distension on blood flow, and that the loss of integrity of the mammary epithelium, which also occurs, is not due simply to mechanical rupture. 8. From present and previous studies, a scheme is proposed to account for the control of mammary function after the cessation of milking in late lactation in the goat.

Mammary uptake of insulin from blood was quantified near parturition in nine dairy cows, some of which were milked prepartum to induce prepartum lactogenesis. Milk samples were immunoassayed for concentrations of insulin, insulin-like growth factor I, and insulin-like growth factor II. Cows milked prepartum, but in which prepartum lactogenesis did not occur (denoted as unsuccessful group), had higher arterial concentrations and greater mammary uptake of insulin during several days immediately preceding parturition than did those with prepartum lactogenesis. Although mammary uptake of insulin was measurable before parturition, estimated uptake on postpartum d 4 through 6 was negligible. Colostrum and milk contained very substantial quantities of insulin, insulin-like growth factor I, and insulin-like growth factor II. Cows in which prepartum lactogenesis was induced (denoted as successful group) had lower concentrations of both forms of insulin-like growth factor in prepartum samples of milk. Cows of the successful group also had a decreasing prepartum profile of milk insulin, but the unsuccessful group had an increasing trend until parturition. Milk concentrations of insulin and both forms of insulin-like growth factor declined rapidly after parturition and completion of lactogenesis.

A balance model of milk synthesis was developed. The model balances carbon flow into and out of the gland and generates sufficient energy and reducing equivalents to meet synthetic requirements. Calculated uptakes of glucose, acetate, and betahydroxybutyrate were sufficient to meet gland requirements. Estimated uptakes of essential amino acids were less than output in milk and had to be adjusted to balance the model. Triacylglyceride fatty acid uptake from blood plasma was less than required for milk fat synthesis. A possible uptake of other plasma fatty acids was postulated to balance the model. Availability of glucose and oxidizable substrate suggested 42% of reducing equivalents for fat synthesis were generated in the pentose cycle and the remainder (58%) by isocitrate dehydrogenase. The balance model was used to formulate a dynamic model with equations to trace fates of isotopically labeled carbons from a variety of substrates. These were used to evaluate effects of changing nutrient availability and rate constants upon patterns of tracer distribution in products and to identify experimental data required to define rate constants uniquely in the model. Additional data are required to define equations for a number of key reactions that exhibit nonlinear kinetics in vivo.

In this paper we examine the ability of the mammary gland to remove from circulating blood three compounds which differ in their physico-chemical and structural properties. Mammary extraction of progesterone, oestrone sulphate and epidermal growth factor (EGF) is similar at peak lactation in goats, but the proportion of labelled infusate that is transferred into milk is greater for oestrone sulphate and EGF than progesterone which is rapidly metabolised by mammary tissue. The kinetics of transfer of progesterone, oestrone sulphate and EGF from blood into milk show that transcellular processes are involved, and on the basis of earlier hypotheses and new information reported here the results indicate the probable importance of simple and facilitated diffusion pathways for progesterone and oestrone sulphate, and secretory mechanisms for oestrone sulphate and EGF. Although evidence is lacking for a direct effect of hormones in milk on mammary function, their concentration in milk may reflect changes in local regulation of mammary secretion. Considerable practical value is attached to the immunodiagnostic use of milk hormone concentrations to determine ovarian and placental endocrine activity during pregnancy in domestic ruminants.

Lactation causes increases in mammary blood flow, gastrointestinal blood flow and cardiac output in the rat, and the suckling-induced release of lactogenic anterior pituitary hormones probably causes these changes, directly or indirectly. Metabolic requirements of lactation in women are similar to those in other animals not artificially selected for a high milk yield but the physiological control of the relationship between milk secretion and nutrient availability is not well understood. The unusual composition of the aqueous phase of human milk (rich in lactose but poor in sodium and potassium ions) can be explained by the same basic mechanisms for secretion of lactose and ions as operate in other animals but with quantitative differences.

When goats were milked every hour for 8 h without the use of exogenous oxytocin, milk ejection each hour appeared to be more complete than had previously been reported. Moreover, the rate of milk secretion increased by 15% over the period of hourly milking. There were no major changes in the composition of the aqueous phase of milk. Mammary blood flow was not apparently changed when goats were milked hourly: there was no significant effect of time of day (between 09.00 and 17.00 h) on blood flow. These results are discussed in relation to the mechanism by which frequent milking increases the rate of milk secretion.

1. A cannulation technique is described for measuring arteriovenous differences across the lactating-rabbit mammary gland. 2. Analysis of milk obtained before and after surgery shows no effect of cannulation on milk constituents. 3. Results of blood analysis show significant net changes in the concentrations of glucose, acetate, 3-hydroxybutrate, triacylglycerols and non-esterified fatty acids across the mammary gland. 4. The molar proportions of individual fatty acids in both the triacylglycerol and non-esterified fatty acid fractions did not alter between the arterial and venous samples. 5. The extraction rates are compared with those obtained from other species.

In ruminants, mammary supply of substrate varies with rate of mammary blood flow and concentrations of blood substrates. Blood concentrations of most mammary substrates, except acetate and tryptophan, do not vary greatly with feed intake, short term. Fasting reduces mammary blood flow, whereas milking and injection of growth hormone or thyroxine increase flow. It is proposed that the fraction of cardiac output that perfuses the udder of lactating ruminants plays a role in regulation of nutrient partitioning between milk and body tissues. In fed animals this fraction is 15 to 16% of cardiac output, which declines on fasting to 8 to 9% and increases slightly following growth hormone treatment to 17.6%. Following realimentation of fasted cows or goats, mammary blood flow takes several hours to return to normal. Investigation of the mechanism of this response, in terms of the ability of the animal to recognize its nutritional status and partition nutrients accordingly, should prove fruitful to understanding causes of variations of milk production in response to feed quantity and quality. Several substrates show increased mammary arteriovenous difference with increasing blood concentrations. This may reflect differing ratios of blood flow:milk yield. The steep gradient of concentration of substrates across the mammary epithelial cell membrane suggests that a major impediment to substrate supply for milk synthesis is the rate of substrate transport across the membrane.

1. Prostacyclin production in mammary gland of two lactating goats measured as the excretion in milk of 6-ketoprostaglandin F1 alpha (6-KPGF1 alpha) was followed for 16 days before, during and after exogenous administration of recombinant bovine growth hormone (GH). 2. 6-KPGF1 alpha was detected in all milk samples in concentrations ranging from 32-99 pg/ml milk independently of the time of sampling. 3. GH-treatment significantly increased milk yield, the concentration and excretion of 6-KPGF1 alpha in milk. 4. The concentration of milk 6-KPGF1 alpha was positively correlated with milk yield in the high (R2 = 0.35), but not in the low yielding goat (R2 = 0.003). 5. The possible role of prostacyclin as a local vasodilator in the mammary gland of goats is discussed.

1. The effect of milking goats 1-4 times an hour for 3-12 hr on the yield and composition of milk has been studied in fed and fasted animals at all stages of lactation.2. It was essential to inject oxytocin (50-400 m-u. I.V.) just before each milking to remove all the milk already in the udder and then the yield was similar to that obtained on twice daily milking (105 +/- 2.1 S.E.%). There were no significant differences between goats or between the two glands of one goat, even if one had been denervated by autotransplantation. However, the variation from hour to hour was 1.5 times greater than from day to day.3. The claim of Zaks (1964) and Zaks, Natochin, Sokolova, Tanasichuk & Tverskoy (1965) that milking every 15 min always produces a large rise in milk Na and a fall in K and lactose, which is characteristic of alveolar milk, is not substantiated. In high yielding goats milking gently by hand or with a cannula caused a small change in K only, but vigorous hand milking exacerbated this fall and also caused a fall in lactose and a rise in Na and Cl. Still larger changes were produced by using excessively large doses of oxytocin (2500 m-u.) when there was also a rise in citrate and total nitrogen. Hourly milking in goats fasted for 24 hr had the same effect.4. In fasted goats the milk yield fell to 90% within 8 hr and to 56 +/- 2.1% of the previous level by 24 hr. It remained at this level for a further 10-12 hr on twice daily or on hourly milking. The yields of autotransplanted glands usually fell slightly but significantly more than that of the glands in situ. In most goats mammary blood flow was halved but in all animals there were large falls in mammary uptake of glucose, acetate and amino acids and greatly increased uptake of free fatty acids. There were significant differences between fasted goats on hourly milking.5. It is concluded that, in spite of changes in milk composition, milking hourly can be a useful technique for studying milk secretion. The striking effects of a short fast in a lactating animal are emphasized.