Assessing Transition Cow Management in Dairy Cows for Improved Health, Milk Production, Pregnancy and Culling Outcomes.

Understanding of the social environment has the potential to benefit dairy cow welfare and production. Our aim was to evaluate the associations of stocking density, calving density, days spent in a pre-partum group before calving (days spent in close-up, DCU) and the number of days from a pen filling event (addition of new cows to the pre-partum pen) on early-lactation health, production, pregnancy and culling outcomes in dairy cows. Data were gathered from 2780 cows in 2 herds. Herd management and reproductive records were analyzed for cows receiving treatment in the first 30 d of lactation (days in milk, DIM) for clinical mastitis, reproductive tract disease, ketosis, milk fever and displaced abomasum. Principal component analysis was used to account for the relationship between gestation length (GL) and DCU and their association with early lactation disease, milk production, pregnancy and culling outcomes. The effect of stocking density, the number of days from a pen filling event to calving and the calving density experienced by a cow in her week of calving was also evaluated. Causal inference was used to correct for confounding bias due to farm identity. The odds of disease in the first 30 DIM increased with stocking density before calving. A quadratic association was found between the first principal component (PC1), representing the combined effect of GL and DCU, and the odds of disease in multiparous cows only. Early lactation milk production and 305 d milk production in multiparous cows increased with PC1 score. Quadratic relationships were found between stocking density at d 8 to 2 before calving with both early lactation and 305 d milk production in multiparous cows but no associations were found in primiparous cows. Calving density showed a quadratic association with 305 d milk production in primiparous cows. The number of days from the last pen filling event to calving was not associated with changes in milk production. Disease occurrence was negatively associated with both early lactation and 305 d milk yield in multiparous cows but only with early lactation milk production in primiparous cows. The occurrence of disease was associated with a delayed time to pregnancy only in primiparous cows while both disease and being in lactation group ≥ 3 were negatively associated with time to pregnancy in multiparous cows. Week 4 milk (W4MK) was positively associated with reduced time to pregnancy in multiparous cows. For primiparous cows, increasing age at calving was associated with increased culling risk, while being in lactation group ≥ 3 was associated with increased culling risk in multiparous cows. Culling risk decreased with increasing W4MK in all cows. These results suggest that gestation length, time spent in close-up and stocking density are important factors influencing disease incidence in early lactation and subsequent lactation performance but had differing effects on primiparous versus multiparous cows. A better understanding of how pre-partum management factors influence postpartum health and milk production can help farms to plan facilities and organize the day-to-day management of cows and will assist in improving cow welfare and productivity.

Association of days in close up, gestation length, and rumination around time of calving with disease and pregnancy outcomes in multiparous dairy cows.

The purpose of this study was to evaluate the effect of rumination times and days spent in a close-up group before calving (DCU) on early-lactation health and reproductive outcomes in dairy cows. Data were gathered for 719 cows located in a single herd. Herd management and reproductive records were analyzed for cows receiving treatment in the first 30 d of lactation (days in milk; DIM) for clinical mastitis, reproductive tract disease, ketosis, milk fever, and displaced abomasum. Rumination times for each cow were downloaded daily from the herd's automated collar system used to generate heat and health alerts for each cow beginning at 21 d precalving until 14 d postcalving. During the first 30 DIM, 121 cows (18%) developed at least 1 disease-any combination of ketosis (40 cows, 5.9% of total), mastitis (17 cows, 2.5%), metritis (75 cows, 11%), milk fever (17 cows, 2.5%), or displaced abomasum (28 cows, 4.1%); 305 cows (45%) were pregnant again at 100 DIM, and an additional 139 cows (20%) were pregnant at 150 DIM. Principal component analysis was used to determine the relationship between gestation length and DCU and their association with the odds of developing disease in early lactation. We did not find any significant association between precalving rumination time and disease within the first 30 DIM. Higher rumination time in the week before calving was shown to be strongly linked to a shorter time to subsequent pregnancy, whereas rumination times postcalving were not associated with changes in the time to pregnancy. Principal component analysis showed that a curvilinear combination of gestation length and DCU (principal component 1) was significantly associated with changes in disease incidence in the first 30 DIM. Gestation length and time spent in close up are important management factors in reducing the incidence of disease in early lactation, and rumination times around calving may help predict future reproductive outcomes.

Use of early lactation milk recording data to predict the calving to conception interval in dairy herds.

Economic success in dairy herds is heavily reliant on obtaining pregnancies at an early stage of lactation. Our objective in this study was to attempt to predict the likelihood of conception occurring by d 100 and 150 of lactation (days in milk, DIM) by Markov chain Monte Carlo analysis using test day milk recording data and reproductive records gathered retrospectively from 8,750 cows from 33 dairy herds located in the United Kingdom. Overall, 65% of cows recalved with 30, 46, and 65% of cows conceiving by 100 DIM, 150 DIM, and beyond 150 DIM, respectively. Overall conception rate (total cows pregnant/total number of inseminations) was 27.47%. Median and mean calving to conception intervals were 123 and 105 d, respectively. The probability of conception by both 100 DIM and 150 DIM was positively associated with the average daily milk weight produced during the fourth week of lactation (W4MK) and protein percentage for test day samples collected between 0 to 30 and 31 to 60 DIM. Butterfat percentage at 0 to 30 DIM was negatively associated with the probability of conception by 100 DIM but not at 150 DIM. High somatic cell count (SCC) at both 0 to 30 and 31 to 60 DIM was negatively associated with the probability of conception by 100 DIM, whereas high SCC at 31 to 60 DIM was associated with a reduced probability of conception by 150 DIM. Increasing parity was associated with a reduced odds of pregnancy. Posterior predictions of the likelihood of conception for cows categorized as having "good" (W4MK >30kg and protein percentage at 0 to 30 and 31 to 60 DIM >3.2%) or "poor" (W4MK <25kg and protein percentage at 0 to 30 and 31 to 60 DIM <3.0%) early lactation attributes with actual observed values indicated model fit was good. The predicted likelihood of a "good" cow conceiving by 100 and 150 DIM was 0.39 and 0.57, respectively (actual observed values 0.40 and 0.59). The corresponding values for a "poor" cow were 0.28 and 0.42 (actual observed values 0.26 and 0.37). Predictions of the future reproductive success of cows may be possible using a limited number of early lactation attributes.

Comparison of three reproductive management strategies for lactating dairy cows using combination of estrus detection or ovulation synchronization and Fixed-Timed Artificial Insemination.

The objective of this study was to compare the reproductive performance of lactating dairy cows submitted to first AI after combination of estrus detection and fixed timed AI (FTAI) and FTAI only. Cows were randomly assigned to receive AI at detected estrus between 50 and 70 d in milk (DIM), if not detected in estrus, were enrolled in either Ovsynch (ED-Ov, n = 485) or PRIDsynch (ED-PR, n = 505) protocols; or received FTAI at 80 DIM after Double-Ovsynch protocol (DO, n = 501). Cows were body condition scored (BCS) at calving and at 43 DIM; and evaluated for postpartum disorders within 7 d postpartum; clinical mastitis, lameness and bovine respiratory disease were recorded until first AI. Ovarian cyclicity was monitored at 43 and 50 DIM, and at 70 and 77 DIM. Pregnancy diagnoses (PD) were performed at 32 and 63 d after AI. Overall prevalence of postpartum anovulation was 7.8%. Pregnancy per AI (P/AI) did not differ between reproductive strategies at 32 d PD (ED-Ov = 43.2%; ED-PR = 41.7%; DO= 45.3%). Primiparous cows had greater P/AI than multiparous cows (53.7% vs 36.8%). Cows on farm 1 had lower P/AI compared with their counterparts on farm 2 (42.1% vs 45.4%). Cows with BCS > 2.5 at 43 DIM had greater P/AI compared with cows with BCS ≤ 2.5 (44.5% vs 34.7%). Similar P/AI for cow's receiving AI at detected estrus and FTAI, low prevalence of disease anovulation may have contributed to the similar performance of ED-Ov, ED-PR and DO.

Effect of pegbovigrastim treatment on the incidence of post-calving antimicrobial treatments in four UK dairy herds.

A randomised controlled trial was carried out in four dairy herds located in the UK to evaluate the effect of pegbovigrastim treatment on the incidence of antimicrobial treatments during the first 30 d of lactation (DIM). Medical treatment records were analysed, and treatments identified where an antibiotic product was used. Records were available for 1865 cows, 933 of which received two injections of pegbovigrastim given approximately 14 d prior to expected calving (IMR) and again within 24 h of calving. 932 cows received no treatment (CON). In total, 11.6% (n = 108/933) IMR cows and 13.2% (n = 123/932) CON cows received at least one antibiotic treatment during the first 30 DIM. Of the IMR cows 2.9% (n = 27/933) were treated with antibiotics for the reason of mastitis along with 3.4% (n = 32/932) of cows from the CON group. 8.9% (n = 83/933) of IMR cows and 10.3% (n = 96/932) of CON cows received antibiotic treatment for a condition other than mastitis, 0.2% (n = 2/933) and 0.8% cows (n = 7/932) from the IMR and CON groups, respectively, received an antibiotic treatment for both mastitis and a reason other than mastitis during the first 30 DIM. Data were analysed with the farm where each cow was located as a random effect and with fixed effects of treatment (IMR or CON), parity (categorised as cows in 1st, 2nd and 3rd or subsequent lactations) and season of calving (autumn [AUT], September through November; winter [WIN], December through February; spring [SPR], March through May; and summer [SUM], June through August), and all 2-way interactions with treatment. Treatment was associated with reduced risk of receiving antibiotic therapy in the first 30 DIM (odds ratio [OR], 0.51; 95% confidence interval [95% CI], 0.28 to 0.94), but a treatment × farm interaction was detected. Compared with IMR, CON cows were more likely to receive an antibiotic treatment on 3/4 farms during the first 30 DIM. However, CON cows on Farm 2 were less likely to do so (12.4% [n = 45/364] vs.15.5% [n = 36/232]). Cows in the third or subsequent lactation were also found to be at increased risk of receiving antibiotic therapy (OR = 1.54; 95% CI, 1.09 to 2.20) than cows in their first lactation. Pegbovigrastim treatment pre-calving may be useful in some herds for reducing the incidence of antimicrobial treatments during early lactation.

Signaling in the yeast pheromone response pathway: specific and high-affinity interaction of the mitogen-activated protein (MAP) kinases Kss1 and Fus3 with the upstream MAP kinase kinase Ste7.

Kss1 and Fus3 are mitogen-activated protein kinases (MAPKs or ERKs), and Ste7 is their activating MAPK/ERK kinase (MEK), in the pheromone response pathway of Saccharomyces cerevisiae. To investigate the potential role of specific interactions between these enzymes during signaling, their ability to associate with each other was examined both in solution and in vivo. When synthesized by in vitro translation, Kss1 and Fus3 could each form a tight complex (Kd of approximately 5 nM) with Ste7 in the absence of any additional yeast proteins. These complexes were specific because neither Hog1 nor Mpk1 (two other yeast MAPKs), nor mammalian Erk2, was able to associate detectably with Ste7. Neither the kinase catalytic core of Ste7 nor the phosphoacceptor regions of Ste7 and Kss1 were necessary for complex formation. Ste7-Kss1 (and Ste7-Fus3) complexes were present in yeast cell extracts and were undiminished in extracts prepared from a ste5delta-ste11delta double mutant strain. In Ste7-Kss1 (or Ste7-Fus3) complexes isolated from naive or pheromone-treated cells, Ste7 phosphorylated Kss1 (or Fus3), and Kss1 (or Fus3) phosphorylated Ste7, in a pheromone-stimulated manner; dissociation of the high-affinity complex was shown to be required for either phosphorylation event. Deletions of Ste7 in the region required for its stable association with Kss1 and Fus3 in vitro significantly decreased (but did not eliminate) signaling in vivo. These findings suggest that the high-affinity and active site-independent binding observed in vitro facilitates signal transduction in vivo and suggest further that MEK-MAPK interactions may utilize a double-selection mechanism to ensure fidelity in signal transmission and to insulate one signaling pathway from another.

Phosphorylation and localization of Kss1, a MAP kinase of the Saccharomyces cerevisiae pheromone response pathway.

Kss1 protein kinase, and the homologous Fus3 kinase, are required for pheromone signal transduction in Saccharomyces cerevisiae. In MATa haploids exposed to alpha-factor, Kss1 was rapidly phosphorylated on both Thr183 and Tyr185, and both sites were required for Kss1 function in vivo. De novo protein synthesis was required for sustained pheromone-induced phosphorylation of Kss1. Catalytically inactive Kss1 mutants displayed alpha-factor-induced phosphorylation on both residues, even in kss1 delta cells; hence, autophosphorylation is not obligatory for these modifications. In kss1 delta fus3 delta double mutants, Kss1 phosphorylation was elevated even in the absence of pheromone; thus, cross-phosphorylation by Fus3 is not responsible for Kss1 activation. In contrast, pheromone-induced Kss1 phosphorylation was eliminated in mutants deficient in two other protein kinases, Ste11 and Ste7. A dominant hyperactive allele of STE11 caused a dramatic increase in the phosphorylation of Kss1, even in the absence of pheromone stimulation, but required Ste7 for this effect, suggesting an order of function: Ste11-->Ste7-->Kss1. When overproduced, Kss1 stimulated recovery from pheromone-imposed G1 arrest. Catalytic activity was essential for Kss1 function in signal transmission, but not for its recovery-promoting activity. Kss1 was found almost exclusively in the particulate material and its subcellular fractionation was unaffected by pheromone treatment. Indirect immunofluorescence demonstrated that Kss1 is concentrated in the nucleus and that its distribution is not altered detectably during signaling.

Glycosylated hemoglobins and glycosylated plasma proteins in the diagnosis of diabetes mellitus and impaired glucose tolerance.

Total and stable glycosylated hemoglobins and glycosylated plasma proteins were determined on 53 patients referred for a glucose tolerance test. Significant correlations were found with fasting blood glucose (r greater than 0.89), 2-h glucose (r greater than 0.69), and area under the glucose tolerance curve (r greater than 0.75), but the correlations with labile glycosylated proteins were not significant. Thirty-one of the patients were normal, five had impaired glucose tolerance (IGT), and seventeen diabetes mellitus (DM) according to the WHO criteria. Comparison of the glycosylated protein values showed that, in all cases, the values for those with IGT and DM were significantly (P less than 0.001) greater than the values for normals. The range of values of stable glycosylated hemoglobins for those with DM (9.4-24.4%), those with IGT (8.6-10.0%), and normals (5.0-8.5%) shows that there was no overlap between overt diabetic subjects and normal subjects. This was also found for total glycosylated hemoglobins. The results for glycosylated plasma proteins, total and stable, were comparable, but one patient with overt DM and two with IGT had values within the normal range. The measurement of glycosylated hemoglobins and glycosylated plasma proteins by the simple, precise, affinity-chromatography method is potentially a quick, accurate, and simple screening test for patients with DM and IGT and deserves consideration as criteria for their diagnosis.

Measurement of glycosylated haemoglobins using an affinity chromatography method.

After removal of the labile material, we have measured the stable glycosylated fraction of haemoglobin with a new, commercially available, phenylboronic acid affinity gel, Glycogel B. The mean value was established for 61 non-diabetics as 7.31 (SD +/- 0.92)% and for 108 diabetics as 12.70 (SD +/- 2.88)%. The method is highly reproducible with a coefficient of variation below 2.0%. The effect of changing the temperature from 7 degrees C to 37 degrees C, and pH from 8.1 to 8.9 was investigated. For accurate results the temperature should be maintained between 20 degrees C +/- 1 degree C, and the pH between 8.6 +/- 0.1. A poor, but significant correlation (r = 0.43) between glycosylated haemoglobin and simultaneous blood glucose was shown. There was a good correlation with the agar gel electrophoretic method (r = 0.95). The slope of the regression line was 1.20 which indicates that this affinity method measures more than just HbA1. The affinity method appears to offer selectivity for diabetics than the electrophoretic method.

Application of analytical methods to automatic analysers.

In adapting methods for automatic analysers modifications to the chemistry may help to simplify machine design. These modifications are acceptable only if they do not adversely influence the accuracy and reproducibility of the method. The strict timing sequence which is ensured in automatic analysers, particularly of the discrete type, allows a simplification of some methods not possible in manual assays.

A study of the hypouricaemic effect of tiaprofenic acid.

Tiaprofenic acid is a non-steroidal anti-inflammatory drug which also has hypouriceamic effect. Studies involving 10 healthy volunteers were designed to investigate the mode of this effect. We postulate that the site of action of tiaprofenic acid is at the cell membrane, the mechanism being an interference with the transport of uric acid from intra- to extra-cellular fluid thus limiting its passage into the plasma. The same mechanism acting on renal tubular cells impedes reabsorption thereby increasing uric acid clearance.

An inexpensive, rapid and precise affinity chromatography method for the measurement of glycosylated haemoglobins.

We have assessed an affinity chromatography technique, using commercially available materials, for the estimation of total glycosylated haemoglobin in the routine clinical chemistry laboratory. The method gives good discrimination between normals (7.31 +/- 0.92%) and diabetics (12.70 +/- 2.88%) and has excellent precision (CV 1.5-2.0%). Labile glycosylated haemoglobin is normally removed as it is so variable. There is no significant correlation between labile glycosylated haemoglobin and blood glucose. Immediate analysis of incubated haemolysates is preferable to storage of haemolysates or erythrocytes. The affinity gel can be reused about 16 times, but oxidation must be reduced by keeping the gel at 4 degrees C in the dark when not in use. The cost of the gel is about 7p a test and 60 samples can be analysed in a working day. The method is not affected by the presence of up to 20% met-haemoglobin and should also give correct values for samples containing genetic variants of haemoglobin.

A sensitive method for the measurement of glycosylated plasma proteins using affinity chromatography.

We describe a simple, sensitive affinity technique for the routine measurement of glycosylated plasma proteins in clinical laboratories. The commercially available phenylboronic acid gel used for the chromatography has recently been marketed as a kit for this purpose (Glycogel Test Kit, Pierce Chemical Co). The manufacturers of this kit recommend loading 200 microliters neat plasma to each 1 ml gel column. This high loading is to enable the direct measurement of protein in the bound and unbound fractions at 280 nm. This loading is consistent with 10-15 mg protein being added per ml gel. Our results show that protein levels greater than 2 mg per ml gel overload the column. Therefore we used a modification of the more sensitive Bradford procedure to measure protein. The method discriminates between normals (6.29 +/- 1.87%) and diabetic patients (12.62 +/- 3.36%) and has good precision (CV 4-6%). The results obtained correlate with the colorimetric method using thiobarbituric acid (r = 0.70) and with glycosylated haemoglobin (r = 0.82).

Whole blood osmolality.

The osmolality of plasma and heparinised whole blood samples collected from hospital patients was estimated using measurement of the depression of freezing point. There was no clinically significant difference between osmolality measurement made on either whole blood, or plasma taken from the same patient. Neither cell volume nor haemolysis was found to affect the measurement. The reproducibility of whole blood measurements was similar to that for determinations carried out on plasma. Measurement of osmolality on whole blood is quicker and cheaper and needs a smaller specimen than if serum or plasma is used.

Sex- and age-specific lungworm infection in Rocky Mountain bighorn sheep during winter.

Sex- and age-specific fecal lungworm (Protostrongylus spp.) larvae concentrations in Rocky Mountain bighorn sheep (Ovis canadensis canadensis) from the Cinnabar Winter Range in southwestern Montana were determined. All 228 fecal samples collected from bighorn sheep of known sex and age class from November to April, 1984 to 1987 contained first-stage lungworm larvae. Fecal lungworm concentrations of ewes and rams declined significantly from late fall through early spring, whereas number of lungworms in lamb feces increased as winter progressed.

Susceptibility of elk to lungworms from cattle.

Two studies were conducted to determine the infectivity of the lungworm, (Dictyocaulus viviparus) of cattle origin, in Rocky Mountain elk (Cervus elaphus nelsoni) or wapiti. In the first study, each of three 9-mo-old elk was administered 3,000 D. viviparus larvae from cattle using a nasogastric tube. In the second study, four 16-mo-old elk were each inoculated with 2,000 D. viviparus from cattle using a nasogastric tube. Elk were observed daily for signs of respiratory disease, and fecal samples were collected during the studies and evaluated for lungworm larvae using a modified Baermann technique. One elk was euthanatized during the patent period for recovery of adult lungworms, and three elk were euthanatized after larvae were no longer detected in feces. Lungworm larvae were not detected before inoculation in any of the 16-mo-old elk, but were detected 22 days after inoculation in one elk, 23 days after inoculation in two elk and 24 days after inoculation in all four elk. The prepatent period of this cattle isolate of D. viviparus in elk is therefore 22 to 24 days. The precise prepatent period was not determined in the three 9-mo-old elk, but larvae were detected in all three elk 25 days after inoculation. Numbers of larvae ranged from 1/ to 101/g feces with peak larval detection occurring 32 to 50 days after inoculation. Elk shed larvae from 22 to 83 days after inoculation, and patent periods of the parasite ranged from 24 to 62 days. Clinical signs of respiratory disease, with the exception of mild coughing after exercise, were not observed during the infections. Results from this experiment indicated that D. viviparus larvae of cattle origin can mature in elk and larvae can be passed in large numbers in feces, but this cattle isolate of D. viviparus was not highly pathogenic in elk.

One-sample pregnancy diagnosis in elk using fecal steroid metabolites.

Recent research has demonstrated the potential of pregnancy diagnosis in elk (Cervus elaphus nelsoni) using immunoassays of fecal steroid concentration. However, multiple samples are required to insure accurate results, limiting its utility for free-ranging animals. We attempted to develop an accurate one-sample pregnancy diagnosis using 153 fecal samples that were collected from free-ranging, radio-collared, adult female elk in Yellowstone National Park (Wyoming, USA) and from captive elk maintained at the Starkey Research Facility (La Grande, Oregon, USA) February through April 1992 and 1997. The pregnancy status of each animal was diagnosed using serum pregnancy-specific protein B (PSPB) assays providing fecal samples from 38 nonpregnant and 115 pregnant animals. Fecal radioimmunoassay (RIA) indicated that mean (+/- SD) progestagens (P4) were elevated significantly in pregnant (2.96 +/- 1.49 micrograms/gm) compared to nonpregnant (0.43 +/- 0.26 microgram/gm) individuals. Confidence intervals (1.96 +/- SE) for the two groups were widely separated (nonpregnant 0.34-0.51, pregnant 2.69-3.24) with little overlap in the range of concentrations measured for each group (nonpregnant 0.09-0.98, pregnant 0.90-8.29). These results indicate that fecal progestagens RIA provides a reliable method of noninvasive pregnancy diagnosis using single fecal samples collected from elk during late gestation. However, independent validation of the suggested discrimination criteria should be performed before routine application.

Antibody response of elk immunized with porcine zona pellucida.

Immunocontraception using porcine zona pellucida (PZP) vaccines is being explored as a nonlethal method of solving the problems of locally overabundant wildlife populations. This study characterized the immunological response of captive elk (Cervus elaphus nelsoni) to PZP challenge using 18 3-yr-old cows and was conducted from 14 September 1994 to 13 December 1995. All animals were given a single PZP inoculation and 1 mo later six of these animals were randomly chosen and received a booster inoculation. Blood samples were drawn from all animals at the time of the initial inoculation and 1, 2, 4, 6, 10, and 15 mo later. Immunological response was assessed by measuring anti-PZP antibody levels in serum. All animals demonstrated a strong immune response with no evidence that the booster enhanced antibody levels. Antibody levels rose from between 0 and 4 at the time of the initial injection to peak levels of 85 to 163 within 2 to 6 mo, followed by a noticeable decline by 15 mo post-vaccination. Limited data suggest that antibody levels > 100 may be required to effect contraception. High individual variability in immune response observed in this study suggests it may be difficult to predict the proportion of animals effectively treated. Disruption of seasonal synchrony in calving also could occur if antibody levels in individuals fall below effective levels while animals are still cycling. These results indicate that immunocontraception using PZP vaccines is possible for elk. However, carefully controlled population experiments will be required in order to assess the potential and limitations for management applications of this technique.