Amino acids activate mTORC1 to release roe deer embryos from decelerated proliferation during diapause.

Embryonic diapause in mammals leads to a reversible developmental arrest. While completely halted in many species, European roe deer (Capreolus capreolus) embryos display a continuous deceleration of proliferation. During a 4-mo period, the cell doubling time is 2 to 3 wk. During this period, the preimplantation blastocyst reaches a diameter of 4 mm, after which it resumes a fast developmental pace to subsequently implant. The mechanisms regulating this notable deceleration and reacceleration upon developmental resumption are unclear. We propose that amino acids of maternal origin drive the embryonic developmental pace. A pronounced change in the abundance of uterine fluid mTORC1-activating amino acids coincided with an increase in embryonic mTORC1 activity prior to the resumption of development. Concurrently, genes related to the glycolytic and phosphate pentose pathway, the TCA cycle, and one carbon metabolism were up-regulated. Furthermore, the uterine luminal epithelial transcriptome indicated increased estradiol-17ß signaling, which likely regulates the endometrial secretions adapting to the embryonic needs. While mTORC1 was predicted to be inactive during diapause, the residual embryonic mTORC2 activity may indicate its involvement in maintaining the low yet continuous proliferation rate during diapause. Collectively, we emphasize the role of nutrient signaling in preimplantation embryo development. We propose selective mTORC1 inhibition via uterine catecholestrogens and let-7 as a mechanism regulating slow stem cell cycle progression.

Estimation of udder emptying based on milk constituents of strip samples after milking.

Milk ejection disorders were induced by oxytocin receptor blockade. We tested the hypothesis that the degree of udder emptying at incomplete milk ejection can be estimated based on the concentration of various milk constituents in different milk fraction samples. To induce different levels of spontaneous udder emptying (SUE) 10 Holstein dairy cows were milked either with or without i.v. injection of the oxytocin receptor blocking agent atosiban (ATO). In ATOearly, 12 µg/kg BW ATO was injected immediately before and in ATOlate directly after a 1-min manual udder preparation. The normal milking routine served as the control treatment. In all 3 treatments the udder was completely emptied by the i.v. injection of 10 IU oxytocin (OT) at the end of spontaneous milk flow. During all experimental milkings 4 milk samples were taken in all treatments: at the start of udder preparation (foremilk; FM), immediately after cessation of spontaneous milk flow and cluster detachment by hand stripping (strip milk; SM), from spontaneous removed milk in bucket 1 (milk before OT; MBOT) and from the milk obtained after OT injection in bucket 2 (milk after OT; MAOT). Fat, protein, lactose, and electrolytes (Na, Cl, and K) were measured in each milk sample. In addition, electrical conductivity (EC) was determined in parallel to continuous milk flow recording. The treatments induced individual degrees of SUE; therefore, the final evaluations of data were based on SUE classes instead of treatments. The most pronounced differences of milk constituents at different degrees of SUE were found for the milk fat content. The fat content of SM and MBOT remained almost unchanged up to 60% SUE, but was considerably higher if >80% of the milk was spontaneously removed. The concentrations of Na and Cl were highest and of K lowest if less than 20% of milk was received in the different samples. The EC was higher in SM and MBOT if <20% of milk was received. In conclusion, the blockade of the OT effect influences primarily the fat content, which confirmed an OT-induced fat secretion during milking. Similar effects are likely found in situations of disturbed milk ejections, caused by a lack of or reduced release of OT in response to different degrees of tactile udder stimulation. Our results show that the measurement of fat content and the EC in SM samples collected after cluster detachment can be used to estimate the completeness of udder emptying.

The mouthpiece chamber vacuum pattern indicates the cessation of milk flow and suits as an indicator to reduce teat end vacuum at a quarter level.

We investigated the suitability of the quarter mouthpiece chamber vacuum (MPCV) as an indicator for cessation of quarter milk flow to potentially adjust the teat end vacuum at a quarter level. We tested the hypothesis that a MPCV increase is a clear indicator of quarter milk flow cessation. In addition, we tested if a quarter-individual vacuum reduction at MPCV increase reduces the mechanical effect on the teat. Ten dairy cows were milked twice daily with a quarter-specific vacuum supply with continuously high (51 kPa; TRT51) or low vacuum setting (41 kPa; TRT41), or high vacuum setting combined with a quarter-specific vacuum reduction by 10 kPa immediately after the quarter-specific MPCV increase (TRT51/41). Whole udder milk flow was continuously recorded. Each treatment was repeated at 4 subsequent milkings. The high vacuum settings (TRT51; TRT51/41) reached higher values in peak flow rate and average milk flow and consequently shorter machine-on time. The time from start of milking until the steep increase of the MPCV was shorter in front than rear quarters, and hence the time from start of MPCV increase until end of milking was longer in front than rear teats. Teat condition of the right front teats was measured for teat wall diameter by ultrasound and teat tissue thickness by cutimeter at 5 and 20 min after each experimental milking. The teat measurements were taken at the teat tip (distal barrel) and 2 cm above the teat tip (proximal barrel). The proximal teat wall diameter tended to be higher in TRT51 than in TRT41, both 5 and 20 min after milking. The distal teat wall diameter at 5 min was greater in TRT51 than in TRT41. In TRT51/41 the teat wall diameter at both locations was intermediate, not significantly different from either TRT51 or TRT41. The distal teat tissue thickness was greater in TRT51 than in TRT41, and tended to be greater in TRT51/41 than in TRT41 at 5 min. The proximal teat tissue thickness at 5 min was higher in TRT51 and TRT51/41 than in TRT41. The teat tissue thickness decreased from 5 to 20 min only in the proximal barrel. The quarter-individual MPCV increase appears to be a suitable indicator of the cessation of milk flow. The lack of a significant reduction of mechanical effect on the teat by a reduced vacuum of 41 kPa indicates that the vacuum level chosen may be still too high under conditions of a separate vacuum supply for each quarter, which prevents a vacuum drop caused by the whole udder milk flow.

Effect of oxytocin use during colostrum harvest and the association of cow characteristics with colostrum yield and immunoglobulin G concentration in Holstein dairy cows.

Our objectives were to determine the effect of oxytocin use during colostrum harvest on colostrum yield and IgG concentration in Holstein dairy cows on a commercial dairy in New York and to describe associations of cow characteristics with these outcomes. Animals were enrolled between July and October 2023 using a randomized block design, with day of enrollment as the unit of randomization. A median (range) of 10 (3-19) cows were enrolled per day. Treatments were (1) 40 IU oxytocin (OXY40), (2) 20 IU oxytocin (OXY20), and (3) an untreated control group (CNTR). Oxytocin was administered i.m. ∼45 s before unit attachment in a rotary parlor. Colostrum weight was measured using the colostrum bucket-embedded scale or a platform scale when the yield was less than the smallest bucket scale. The concentration of IgG ([IgG]) in colostrum was determined using radial immunodiffusion and used to calculate total IgG, and DM (%) was determined by oven drying. Individual cow characteristics such as parity, calf sex, weight, dystocia score, stillbirth, and milk production in wk 4 of lactation; additionally, for multiparous cows, dry period length and previous lactation dry-off linear SCS were collected. Data were analyzed separately for primiparous and multiparous cows using backward stepwise elimination. To produce final mixed effects ANOVA models, primiparous cows (n = 201) were randomized to 35.8% (n = 72) OXY40, 32.8% (n = 66) OXY20, and 31.3% (n = 63) CNTR. Multiparous groups (n = 435) were randomized to 34.7% (n = 151) OXY40, 29.7% (n = 129) OXY20, and 35.6% (n = 155) CNTR. The median (range) colostrum yield was 6.0 (0-20.6) kg and [IgG] was 98.5 (0.1-293.6) g/L in the study population. In primiparous cows, OXY40 had a higher colostrum yield (LSM [95% CI]) of 5.4 (4.9-5.9) kg compared with both OXY20 (4.1 [3.5-4.7] kg) and CNTR (3.8 [3.3-4.3] kg). In multiparous cows, OXY40, OXY20, and CNTR did not differ in colostrum yield (5.9 [5.3-6.5], 5.7 [5.1-6.3], and 5.4 [4.8-6.0] kg, respectively), but colostrum yield was greater in parity 2 compared with all other parities, cows giving birth to male calves, cows with the highest milk production at wk 4 of lactation, and cows with a dry period of >65 d. Oxytocin use did not affect [IgG] in either primiparous or multiparous cows, but [IgG] was highest in cows in parity ≥4 and lowest in cows dry >65 d. In summary, oxytocin use at 40 IU i.m. was associated with a higher colostrum yield in primiparous cows only and no change in [IgG]. Oxytocin use likely addressed disturbed milk ejection and therefore increased colostrum yield in heifers milked for the first time in a rotary parlor. This study confirms cow characteristics associated with colostrum production within a single herd.

Increased pulsation ratio in rear quarters reduces machine-on time and milking on empty teats.

Dairy cows usually store more milk in rear than in front quarters. At conventional machine milking this leads to an earlier cessation of milk flow in front than in rear quarters and hence unavoidable milking on empty front teats. We tested the hypothesis that an increased pulsation ratio in rear quarters reduces both milking on empty front teats and machine-on time. We have also tested if the different amplitude of cyclic vacuum fluctuations during milking with simultaneous vs. alternate pulsation within front and rear quarters, respectively, impacts milking characteristics. Ten Holstein dairy cows were milked twice daily at 14-h and 10-h milking intervals. Pulsation ratio in front quarters was 65:35, but was either set at 65:35, 70:30, 75:25 or 80:20 in rear quarters. Whole udder milk flow, claw vacuum, and mouthpiece chamber vacuum of all 4 quarters were recorded during milking. Teat tissue thickness of all 4 teats was measured with a cutimeter at 5 and 30 min after cluster detachment. Total milk yield did not differ among treatments, indicating complete udder emptying. The time of quarter milk flow (tQMF) of front quarters did not differ among treatments but decreased with increasing pulsation ratio in rear quarters. In addition, the faster milking of rear quarters at higher pulsation ratio caused higher peak and average milk flow rates, a shorter duration of milk flow decline, and a reduced machine-on time. The increased milk flow at a higher pulsation ratio did not cause a significant reduction of the claw vacuum at the used sample size. Teat tissue thickness did not significantly differ among treatments, likely because milking on empty teats did almost not occur in the rear quarters, and treatment settings did not differ in front quarters. Simultaneous pulsation caused higher amplitudes of cyclic vacuum fluctuation than alternate pulsation. However, milking characteristics did not differ between simultaneous and alternate pulsation. In conclusion, an increased pulsation ratio in rear quarters reduces machine-on time, increases milking performance, and reduces the time of milking on empty teats in front quarters, without an increased impact on teat tissue thickness.

The interplay of continuous milk ejection and milking system with and without prestimulation at different vacuum settings.

Efficient machine milking requires an optimal interaction of alveolar milk ejection in the udder and milk removal by the milking machine. The aim of the present study was to test whether the equilibrium between continuous milk ejection and milk removal can also be maintained at very fast milking through a particularly high vacuum. Eight Holstein dairy cows were milked at 42, 52, or 60 kPa, with (PS) or without (nPS) prestimulation. Each of the 6 treatments was conducted at 2 afternoon milkings in each animal. The prestimulation lasted 40 s and consisted of forestripping and teat cleaning. The cluster attachment followed after a 20-s latency period. Throughout each milking, B-mode ultrasound videos of the gland cistern of 1 front quarter as well as milk flow and claw vacuum curves were recorded. Total milk yield was neither affected by nPS or PS nor by the vacuum level. Milk removed within the first minute and the first 2 min of milking and average milk flow were higher, and the duration of incline and time until peak milk flow were shorter at PS than at nPS milkings at all vacuum levels. Machine-on time was shorter at PS than at nPS milkings, although only at 42 and 52 kPa vacuum, obviously caused by the high percentage of bimodalities occurring in nPS milkings (17% bimodalities in PS vs. 92% bimodalities in nPS milkings). The frequency of bimodalities was higher at high than at low vacuum both in PS and nPS milkings. Peak flow rate and average milk flow were both higher at higher vacuum levels. The duration of milk flow plateau was shorter at 60 kPa than at 42 kPa milkings. At the highest vacuum (60 kPa), the shorter plateau phase indicated a declining milk ejection rate toward the end of the plateau phase, and milk ejection could no longer keep up with the fast milk removal; hence, a higher milking efficiency at a higher vacuum level could only be achieved as long as the gland cistern remained sufficiently filled by the continuous milk ejection. The ultrasound imaging confirmed this finding as the duration of cisternal area plateau in the recorded front quarter was shorter at high than at low vacuum. Thus, the highest vacuum of 60 kPa did not cause a shorter machine-on time than 52 kPa. In conclusion, milking at a very high vacuum can increase milking efficiency compared with a low vacuum. However, a vacuum reduction at the start and toward the end of milking is required to prevent overmilking if milking is performed at a very high vacuum.

Effects of different nutrient supply on metabolism and mammary immune response to an LPS challenge in early lactation of dairy cows.

Energy and nutrient deficiency in dairy cows in early lactation is considered to contribute to their increased susceptibility to mastitis. We have tested the hypothesis that feeding diets with high contents of either nitrogenic, glucogenic, or lipogenic components in early lactation affects both the endocrine and metabolic status, as well as the mammary immune competence. After calving, cows were fed increasing amounts of concentrate up to 10 kg/d rich in crude protein (nitrogenic, n = 10), glucogenic precursors (glucogenic, n = 11), or lipids (lipogenic, n = 11). In wk 3, one udder quarter was challenged with lipopolysaccharide (LPS) from Escherichia coli. Blood and milk were sampled on the day before LPS challenge (d -1), and on d 0, 1, 2, 3, and 9 after LPS challenge. On the day of LPS challenge additional samples were taken hourly for quarter milk and every 3 h for blood. Urea concentrations were higher in plasma and milk of cows fed the nitrogenic diet. However, plasma concentrations of glucose, cholesterol, triglycerides, ß-hydroxybutyrate, nonesterified fatty acids, as well as insulin, glucagon, and insulin-like growth factor-1 were not affected by the different diets. The mammary immune challenge induced a substantial increase of somatic cell count (SCC) in the treated quarter, and a transient decrease of total milk yield and white blood cells similar in all diet groups for one day. The absolute phagocytosis of blood leukocytes was decreased; however, the phagocytosis per cell was increased in glucogenic-fed cows at 6 h after LPS challenge. During mammary inflammation an insulin resistance, shown by increased plasma glucose, insulin, and glucagon, developed similarly in all diet groups. ß-hydroxybutyrate and nonesterified fatty acids were decreased at 1 d after LPS challenge in glucogenic-fed cows only. Cholesterol did not change, and triglycerides only decreased significantly in lipogenic-fed cows 6 h after challenge. On d 9 after LPS challenge, SCC and milk yield and metabolic factors were recovered in all groups. In conclusion, the endocrine and metabolic situation, and the immune response to intramammary LPS of dairy cows during early lactation was not substantially influenced by the elevated supply of nitrogenic, glucogenic, or lipogenic components due to the provided feed in this study.

Effects of feeding level, milking frequency, and single injection of cabergoline on blood metabolites, hormones, and minerals around dry-off in dairy cows.

This study aimed to investigate the effect of the different dry-off strategies based on reducing feeding level (normal vs. reduced energy density), reducing milking frequency (twice vs. once daily), and administration of a dopamine agonist after last milking (i.e. saline vs. cabergoline injection) on blood metabolites, hormones, and minerals around dry-off. In this experiment, 119 Holstein dairy cows were used in a 2 × 2 × 2 factorial arrangement. In the last week before dry-off, cows were allocated to 1 of the 4 possible dry-off strategies based on feeding level and milking frequency. Within 3 h after last milking, cows were injected with either saline or a D2 dopamine agonist (cabergoline; Velactis, Ceva Santé Animale, Libourne, France; labeled for use only with abrupt dry-off, e.g., no preceding reduction in feeding level or milking frequency before last milking). After dry-off, all cows were fed the same dry cow diet and data collection continued for a week. Blood samples were collected from the coccygeal vein on d -9, -6, -5, -2, 1, 2, 5, and 7 relative to dry-off. Additionally, blood was sampled at 0, 3, and 6 h relative to injection of either cabergoline or saline, equivalent to d 0.125, 0.250, and 0.375 relative to last milking (dry-off). The reduced feeding level before dry-off caused reduced glucose and insulin concentrations as well as increased free fatty acid concentrations, particularly when reduced feeding level was combined with milking the cows 2× daily. The intramuscular injection of cabergoline caused the expected reduction in circulating prolactin concentrations. In addition, dopamine-agonist cabergoline induced an atypical simultaneous pattern of plasma metabolites (i.e., increased glucose and free fatty acid concentrations), hormones (i.e., reduced insulin and increased cortisol concentrations), and minerals (i.e., reduced calcium concentration), indicating that normal metabolic and mineral homeostatic regulations were hindered after the injection of ergot alkaloid cabergoline. In conclusion, reducing milking frequency seems the best management strategy to reduce milk production at dry-off among those tested in this study.

Inhibition of lipopolysaccharide-induced suppression of luteal function in isolated perfused bovine ovaries.

Recently, we observed that lipopolysaccharide (LPS) suppresses corpus luteum (CL) function in isolated perfused ovaries. It remained unclear if this suppression was due to increased luteal PGF2α secretion or LPS-induced apoptosis. Therefore, possible impacts of PGF2α and LPS were inhibited by a non-steroidal anti-inflammatory drug (flunixin) and an endotoxin-binding agent (polymyxin B), respectively. Bovine ovaries with a mid-cycle CL were collected immediately after slaughter and perfused for 240 min. After 50 min of equilibration, either flunixin or polymyxin B (5 µg/ml of each) were added to the perfusion medium of six ovaries, respectively. All ovaries (n = 12) were treated with E. coli LPS (0.5 µg/ml) 60 min after the onset of perfusion, and received 500 I.U. of hCG after 210 min of perfusion. Progesterone and PGF2α were measured in the effluent perfusate every 10 and 30 min, respectively. Biopsies of the CL were collected every 60 min to determine the mRNA expression of the cytokine TNFA and factors of apoptosis (CASP3, -8). Flunixin-treatment inhibited the increase of PGF2α after LPS-challenge that was observed in the polymyxin B-treated (PX-LPS) ovaries. After hCG-stimulation, progesterone secretion increased (P < 0.05) in group PX-LPS but not in the flunixin-treated (F-LPS) ovaries. Compared to initial values before LPS-challenge, luteal mRNA expression of TNFA and CASP3 was increased (P < 0.05) in group F-LPS at 120 and 180 min, respectively, and those of CASP8 was decreased (P < 0.05) in PX-LPS at 60 and 120 min after LPS-treatment. In conclusion, although flunixin managed to inhibit PGF2α, it did not suffice to successfully prevent LPS-induced apoptosis. However, endotoxin-binding polymyxin B resulted in luteal responsiveness to hCG after LPS-challenge.

Reduced liner-open phase and vacuum instead of prestimulation increase parlor efficiency in dairy cows.

Adequate prestimulation is considered a requirement for a fast, gentle, and complete udder emptying at machine milking. Reduced vacuum or reduced liner-open phase of pulsation (or both) may replace prestimulation and consequently reduce work load of the milker and increase parlor efficiency. In the present study we compared 2 milking routines (MR) with manual prestimulation (LPrep = long preparation: 15 s of forestripping, teat cleaning, and stimulation; SPrep = short preparation: 5 s of teat cleaning) followed by 1-min latency period and milking at standard vacuum and pulsation settings (claw vacuum 44 kPa, pulsation rate 60 cycles/min, pulsation ratio 65/35) with 2 MR consisting of 5 s of teat cleaning, immediate cluster attachment and milking at reduced vacuum with or without shortened liner-open phase of pulsation until milk flow exceeded 400 g/min (RP = reduced pulsation: pulsation ratio 30/70, pulsation rate 50 cycles/min, claw vacuum 44 kPa; RPV = reduced pulsation and vacuum: pulsation ratio 30/70, pulsation rate 50 cycles/min, claw vacuum 34 kPa). Cluster detachment was performed at 400 g/min in all MR. Ten Holstein dairy cows were milked twice daily at 14-h and 10-h milking intervals. Milk flow and electrical conductivity (EC) were recorded throughout milking. During the first 2 min of each milking ultrasound cross-section images of the gland cistern of one front quarter were recorded, and 5 min after the end of milking teat tissue thickness of both front teats was measured by using a cutimeter. Most milking characteristics such as total milk yield, average milk flow, and machine-on time reached higher values at 14-h than 10-h milking intervals, but did not differ among MR. However, the occupancy time (time from the first touch of the udder until cluster detachment) was considerably shorter in RP and RPV compared with LPrep and SPrep. Ultrasound cross section areas of the gland cistern were larger in LPrep than in RP and RPV indicating that milk ejection already occurred at cluster attachment in LPrep. This assumption is also supported by the lower EC at cluster attachment in LPrep than in RP and RPV, which was caused by the presence of alveolar milk in the gland cistern after milk ejection. The MR RP and RPV increase parlor efficiency and are work-saving alternatives to MR, which include an adequate prestimulation (LPrep). However, shortening prestimulation to a 5-s teat cleaning followed by a latency period and milking at regular vacuum and pulsation is not adequate to save occupancy time. Because milking was performed at a relatively low vacuum (44 kPa) and at a detachment level of 400 g/min, teat tissue thickness did not differ among MR, and the vacuum reduction in RPV did not cause an additional advantage for teat condition compared with RP.

Drying-off practices on Swiss dairy farms: Status quo and adoption potential of integrating incomplete milking.

Drying-off practices to reduce milk production before dry-off are gaining attention because high milk yields at dry-off are becoming more common and increase the risk to cow health and welfare during the dry period. Incomplete milking for the last days before dry-off is one approach for reducing milk production. We conducted an online survey to determine the currently used drying-off practices on Swiss dairy farms and to identify the adoption potential of integrating incomplete milking before dry-off. In March 2021, the online survey was sent to a representative sample of 1,974 Swiss dairy farmers. A total of 518 completed questionnaires were analyzed. The mean number of dairy cows per farm was 39 (range: 11-140 cows). Thirty-five percent of cows produced considerable quantities of milk (>15 kg/d) at dry-off, and milk yield at dry-off increased with increasing annual milk yield. Abrupt dry-off was applied on 45% of the farms. The participants reported observing behavioral changes of cows such as increased vocalizations and decreased lying time associated with dry-off. Selective dry cow therapy was applied on 74% of the farms, and 44% of the participants indicated the use of antibiotics at dry-off as being "rather often," "often," or "always." Correlation analysis revealed that with increasing annual milk yields, the frequency of observed behavioral changes and antibiotic use at dry-off increased as well. Therefore, drying-off approaches that reduce milk production while supporting cow welfare are needed. We found that farmers showed an interest in testing the presented drying-off approach of incomplete milking. In addition, the farmers indicated that they would be more willing to test incomplete milking before dry-off if it became available for automated use in milking parlors or robots. Uncertainties regarding udder health appeared to be the main barrier for the adoption potential of this approach.

The teat cup detachment level affects milking performance in an automatic milking system with teat cleaning and milking in the same teat cup.

The goal of the present study was to determine the best quarter milk flow for teat cup detachment depending on lactational stage and milking interval to optimize the milking process in automatic milking systems (AMS). Milking characteristics and post-milking teat condition were recorded in an AMS with all actions from teat cleaning to post-milking teat dipping occurring in the same teat cup and liner (GEA DairyRobot R9500). In 24 dairy cows, 12 in early (<80 DIM) and 12 in late lactation (>180 DIM), 294 milkings were recorded during 12 consecutive days. Teat cup detachment was set at a quarter milk flow of 50, 87.5 or 125 g/min. Voluntary milking intervals varied similarly in cows in early (6.1-14.8 h) and late lactation (6.3-15.7 h). Total milk yield, milk production per h and average milk flow were higher in early than in late lactation cows. Total milk yield per milking did not differ between detachment levels. The mean milking time was reduced by up to 1.5 min at the highest compared to the lowest detachment level mainly in early lactation cows (P < 0.05). However, no significant effect of the detachment level on milking time was observed at milking intervals >10 h. Average milk flow was higher at milking intervals >10 h than ≤10 h (P < 0.05). In the early lactation cows (only) the average milk flow increased with higher detachment levels (P < 0.05). Teat condition did not differ among detachment settings or milking intervals. In conclusion, teat cup detachment up to 125 g/min reduces milking time in both early and late lactation without a loss of milk yield or affecting the teat condition. Combined with a higher average milk flow through avoiding too short milking intervals, the total milking time and hence stall occupancy can be optimized by early teat cup detachment.

Effects of short and long milking intervals on milking characteristics and changes of milk constituents during the course of milking in crossbred Istrian × Awassi × East-Friesian ewes.

The main objective of this experiment was to evaluate the effects of two milking intervals (8 and 16 h) on milk constituents (fat, protein, lactose, dry matter, and log10 SCC) of nineteen Istrian × Awassi × East-Friesian crossbred ewes in different milk fractions (0-25, 25-50, 50-75 and 75-100%) during the course of milking and in machine stripping (MS) milk. Furthermore, we sought to determine the effect of the two milking intervals on milking characteristics (average milk flow rate, peak milk flow rate, machine-on time, total milk yield, and milk production rate) and whether each milk constituent within each milking interval is best described by a linear, quadratic, or cubic function. Average milk flow rate and milk yield per milking decreased in the 8 h milking interval compared to the 16 h milking interval (P < 0.05). Peak milk flow rate, machine-on time, and milk production rate were not different between the two milking intervals. Overall, milk fat content, dry matter content, and log10 SCC increased in the 8 h milking interval compared to the 16 h milking interval (P < 0.05). Milk protein content did not change through the main milk fractions at either milking interval. Milk lactose content did not change through the milk fractions at the 8 h milking interval, whereas it decreased in the 75-100% and stripping milk fractions at the 16 h milking interval (P < 0.05). The 0-25% and stripping milk fractions contained the highest log10 SCC compared to all other milk fractions (P < 0.05). Changes of milk fat and dry matter content throughout milking were best described by quadratic functions, whereas milk protein content, milk lactose content, and log10 SCC were best described by different functions depending on the milking interval. These results demonstrate that milking interval influenced all milk constituents in various milk fractions during the course of ewe milking. Moreover, milking characteristics such as average milk flow and total milk yield, and the appropriate mathematical function to characterize milk constituents throughout a milking, were affected by milking interval.

Colostrogenesis: Role and Mechanism of the Bovine Fc Receptor of the Neonate (FcRn).

Colostrogenesis is a separate and unique phase of mammary epithelial cell activity occurring in the weeks before parturition and rather abruptly ending after birth in the bovine. It has been the focus of research to define what controls this process and how it produces high concentrations of specific biologically active components important for the neonate. In this review we consider colostrum composition and focus upon components that appear in first milked colostrum in concentrations exceeding that in blood serum. The Fc Receptor of the Neonate (FcRn) is recognized as the major immunoglobulin G (IgG) and albumin binding protein that accounts for the proteins' long half-lives. We integrate the action of the pinocytotic (fluid phase) uptake of extracellular components and merge them with FcRn in sorting endosomes. We define and explore the means of binding, sorting, and the transcytotic delivery of IgG1 while recycling IgG2 and albumin. We consider the means of releasing the ligands from the receptor within the endosome and describe a new secretion mechanism of cargo release into colostrum without the appearance of FcRn itself in colostrum. We integrate the insulin-like growth factor family, some of which are highly concentrated bioactive components of colostrum, with the mechanisms related to FcRn endosome action. In addition to secretion, we highlight the recent findings of a role of the FcRn in phagocytosis and antigen presentation and relate its significant and abrupt change in cellular location after parturition to a role in the prevention and resistance to mastitis infections.

Type of teat cup liner and cluster ventilation affect vacuum conditions in the liner and milking performance in dairy cows.

The optimal milking cluster should milk as gently as possible to minimize the mechanical effect on the teat tissue at an optimal milking performance and milk quality. The objective of this study was to investigate the influence of liner shape (round vs. triangular) and type of cluster ventilation (claw vs. mouthpiece chamber; MPC) on milking performance and vacuum at the teat end and in the MPC. Our hypothesis was that liner shape and cluster ventilation affect milking performance and MPC vacuum. Six Holstein Friesian cows were milked twice daily over 12 d with a bucket milker, using 4 different cluster types that combined liner shape and type of cluster ventilation at 3 different system vacuum settings (35, 42, and 50 kPa) in an incomplete randomized block design. Milk flow and vacuum in the MPC, at the teat end (measured in the short milk tube), and in the short pulse tube were continuously recorded during milking. Milk flow was higher, and hence main milking time was shorter, with the round than with the triangular liners. The MPC vacuum was lower in round than triangular liners, which was caused by higher air leakage between teat and liner barrel in the triangular liners. The MPC vacuum, as well as its cyclic fluctuations, increased at the end of milking (immediately before cluster detachment) in all cluster types, with the highest amplitude of fluctuation in the triangular liners with MPC ventilation. The MPC ventilation reduced the MPC vacuum in both liner types at the end of milking, and also in triangular liners during peak milk flow. Despite the observed differences of MPC vacuum, the ventilation type did not affect milking performance. However, milking with triangular MPC-ventilated liners caused an increased proportion of foamed milk, which could potentially have a negative effect on milk quality.

Increased teat wall thickness in response to machine milking.

Transiently increased teat wall thickness in response to machine milking has been documented by various methods, including ultrasound. However, correlative ultrasonography and histology to detect the origin of this phenomenon is lacking. The first goal of the present study was to evaluate and compare milking-related changes of the teat tissue in 2 breeds of dairy cows (11 Simmental and 3 Holstein) using B-mode ultrasonography. Additionally, the observed changes were compared with ultrasonographic findings in a Holstein cow with periparturient udder edema. Finally, corresponding histological sections of the Simmental teats were analyzed and compared with those from a lactating nonmilked Angus cow. We hypothesized that the mechanical load of both stretching by the vacuum during phases of open teat cup liner and compression by the closed liner during machine milking results in a transient congestion of blood vessels in the teat wall. The barrel of 1 front teat of each cow was scanned immediately before and after machine milking (system vacuum: 42 kPa; pulsation rate: 60 cycles/min; pulsation ratio: 65:35). Shortly after milking (33 ± 6 min), the Simmentals were slaughtered, and their scanned teat was immediately removed and processed for investigation by light microscopy. Ultrasonography after milking revealed anechoic tubular structures mainly in the inner half of the teat wall. Histological examination revealed these structures to be thick-walled veins. The left front and hind teats of the nonmilked lactating cow, collected and prepared identically to those from the Simmental cows, showed the same histological features. Ultrasonographic measurements showed that the diameter of these veins significantly increased after milking compared with matching images before milking. This effect was most pronounced in the Holstein cows. Similarly, these veins were very prominent in the periparturient cow. However, neither the milked cows, including the periparturient cow, nor the lactating nonmilked cow provided any evidence of edematous extravasation on ultrasonography or histology. These findings corroborated our hypothesis that the increase in size of thick-walled veins in the teat tissue is the main reason for the thickening of the teat walls in response to machine milking.

Influence of nutrient availability on in vitro growth of major bovine mastitis pathogens.

The aim of the present study was to investigate the effects of milk composition changes on the in vitro growth of bovine mastitis pathogens. Nutritional requirements of three major bovine mastitis pathogens Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Streptococcus uberis (S. uberis) were investigated in vitro. We used ultra-high temperature (UHT) treated milk with different contents of fat, protein, and carbohydrates to test the influence of the availability of various milk constituents on pathogen growth characteristics. Additionally, the bacterial growth was investigated under experimentally modified nutrient availability by dilution and subsequent supplementation with individual nutrients (carbohydrates, different nitrogen sources, minerals, and different types of B vitamins) either to milk or to a conventional medium (thioglycolate broth, TB). Varying contents of fat, protein or lactose did not affect bacterial growth with the exception of growth of S. uberis being promoted in protein-enriched milk. The addition of nutrients to diluted whole milk and TB partly revealed different effects, indicating that there are media-specific growth limiting factors after dilution. Supplementation of minerals to diluted milk did not affect growth rates of all studied bacteria. Bacterial growth in diluted whole milk was decreased by the addition of high concentrations of amino acids in S. aureus, and by urea and additional B vitamins in E. coli and S. aureus. The growth rate of S. uberis was increased by the addition of B vitamins to diluted whole milk. The present results demonstrate that growth-limiting nutrients differ among pathogen types. Because reduced bacterial growth was only shown in diluted milk or TB, it is unlikely that alterations in nutrient availability occurring as a consequence of physiological changes of milk composition in the cow's udder would directly affect the susceptibility or course of bovine mastitis.

Plasma cholesterol levels and short-term adaptations of metabolism and milk production during feed restriction in early lactating dairy cows on pasture.

Low plasma total cholesterol (TC) concentrations are characteristic during the negative energy balance in early lactating dairy cows. The objective was to investigate short-term effects of different TC concentrations during an aggravated energy deficiency through a 1-week concentrate withdrawal on adaptations of metabolism and milk production. Multiparous Holstein cows (n = 15) were investigated during 3 week beginning at 24 ± 7 DIM (mean ± SD). Cows were kept on pasture and received additional concentrate in experimental week 1 and 3, while in week 2, concentrate was withdrawn. Blood was sampled once and milk twice daily. Based on their average TC concentration during week 1 (prior to concentrate withdrawal), cows were retrospectively assigned into a high (H-Chol; n = 8, TC ≥ 3.36 mmol/L) and a low TC groups (L-Chol; n = 7, TC < 3.36 mmol/L). Concentrations of phospholipids and lipoproteins were higher in H-Chol compared to L-Chol throughout the study (p < 0.05). During concentrate withdrawal, milk yield, glucose and insulin concentrations decreased similarly in both groups, while milk fat, milk acetone and plasma BHB were higher in H-Chol compared to L-Chol (p < 0.05). Compared to initial values, plasma NEFA, TAG and VLDL increased in both groups within 2 days after concentrate withdrawal (p < 0.05). Concentrations of NEFA during week 2 were greater in L-Chol compared to H-Chol (p < 0.05). Despite reintroduction of concentrate, milk yield in H-Chol remained lower for two more days compared to week 1 (p < .05), whereas milk yield recovered immediately in L-Chol. Activity of aspartate aminotransferase was higher in H-Chol compared to L-Chol in week 2 (p < 0.05). Greater plasma TC concentrations were associated with a reduced increase of NEFA. Further research is warranted if TC concentrations are related to adipose tissue mobilization and fatty acid turnover.

Different vacuum levels, vacuum reduction during low milk flow, and different cluster detachment levels affect milking performance and teat condition in dairy cows.

Traditionally, machine milking is performed at a constant vacuum supply. The system vacuum has to be set high enough to allow a sufficiently high vacuum at the teat end, despite the inevitable vacuum drop caused by milk flow. This leads to an increased vacuum load on the teat, especially when milk flow ceases at the end of milking. We tested the hypothesis that a milk flow-controlled adaptation of vacuum settings during milking allows even higher vacuum levels than are usually recommended during the period of high milk flow if the vacuum is reduced during low milk flow. Combined with a high cluster detachment flow rate level, increased milking performance is expected without an increased effect on teat tissue. Ten Holstein dairy cows were milked with a bucket milker with the claw vacuum adjusted in the absence of milk flow at a regular (43 kPa) and high (48 kPa) claw vacuum, with and without vacuum reduction during low milk flow (<2 kg/min), and combined with different cluster detachment levels (0.2, 0.6, and 1 kg/min). Each treatment was applied in each cow during 4 subsequent milkings in a randomized crossover design. Both claw vacuum and milk flow were continuously recorded throughout milking. Teat tissue thickness was measured using a cutimeter and teat wall diameter was measured by B-mode ultrasonography at 5 min after the end of milking. Milk yield was not affected by either vacuum settings or detachment levels. Machine-on time in treatments with vacuum reduction was shorter at high than at low vacuum and decreased with increasing detachment levels. Average milk flow was higher at high than at low vacuum and reached highest values in milkings without vacuum reduction at both vacuum levels. The average milk flow was higher at a cluster detachment of 1 kg/min than at 0.2 kg/min. However, both teat tissue thickness and (as a tendency) teat wall diameter at 5 min after cluster detachment were higher in milkings at high vacuum without vacuum reduction compared with all other treatments. In conclusion, high claw vacuum up to 48 kPa increases milking performance because of higher milk flow and reduced machine-on time. Negative effects of high vacuum on teat tissue are prevented by reducing vacuum during low milk flow (<2 kg/min) at the start and end of milking. Additionally, using a high cluster detachment level reduces machine-on time without a loss of harvested milk.

Short communication: Teat wall diameter and teat tissue thickness in dairy cows are affected by intramammary pressure and by the mechanical forces of machine milking.

The objective of this study was to validate and apply 2 different methods to record changes in teat tissue related to machine milking. Teat wall diameter was measured via B-mode ultrasound cross sectioning with a 7.5-MHz linear probe. Teat tissue thickness was measured using a cutimeter (spring-loaded caliper, spring constant 6.5 N/cm, 0.5 N at closed jaws). Both methods were applied at the teat barrel, 2 cm above the teat tip. In experiment 1, 24 teats from freshly slaughtered cows were used to perform ultrasound imaging (12 teats) or cutimeter measurements (12 teats) while the teat cisterns were filled with water to increase the intracisternal pressure from 0 to 30 kPa in steps of 1 kPa. Teat tissue thickness did not change at an intracisternal pressure from 0 to 10 kPa but increased with intracisternal pressure at levels >10 kPa. In contrast, teat wall diameter decreased with intracisternal pressure between 0 and 7 kPa but did not significantly change at a pressure ≥7 kPa up to 30 kPa. Significant Pearson correlation coefficients between intracisternal pressure and teat wall diameter were observed from 0 to 7 kPa (r = -0.38), and between intracisternal pressure and teat tissue thickness from 10 to 30 kPa (r = 0.45). In experiment 2, ultrasound and cutimeter measurements were performed in 12 lactating Holstein cows. Measurements before and during milking, immediately after cluster removal, with normal milking or with a 5-min overmilking, were performed and continued at 5-min intervals for 60 min and at 10-min intervals until 120 min. Additionally, with the 5-min overmilking treatment, measurements were continued at 60-min intervals up to 10 h after milking. Teat wall diameter decreased in response to milk ejection, followed by a continuous increase during the course of milking, with highest values after 5 min overmilking. Teat tissue thickness did not change during milking but was significantly increased after overmilking. Teat wall diameter and teat tissue thickness recovered to premilking levels within 35 min after normal milking and within 60 min after overmilking. Until 10 h after overmilking, the teat wall diameter decreased steadily, whereas teat tissue thickness was unfluctuating. In the physiologically relevant range of intramammary pressure, ultrasound measurements of the teat wall were affected by both intramammary pressure and mechanical forces, whereas cutimeter measurements were not affected by the intramammary pressure.