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.

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.

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.

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.

Replication factors MCM2 and ORC1 interact with the histone acetyltransferase HBO1.

The minichromosome maintenance (MCM) proteins, together with the origin recognition complex (ORC) proteins and Cdc6, play an essential role in eukaryotic DNA replication through the formation of a pre-replication complex at origins of replication. We used a yeast two-hybrid screen to identify MCM2-interacting proteins. One of the proteins we identified is identical to the ORC1-interacting protein termed HBO1. HBO1 belongs to the MYST family, characterized by a highly conserved C2HC zinc finger and a putative histone acetyltransferase domain. Biochemical studies confirmed the interaction between MCM2 and HBO1 in vitro and in vivo. An N-terminal domain of MCM2 is necessary for binding to HBO1, and a C2HC zinc finger of HBO1 is essential for binding to MCM2. A reverse yeast two-hybrid selection was performed to isolate an allele of MCM2 that is defective for interaction with HBO1; this allele was then used to isolate a suppressor mutant of HBO1 that restores the interaction with the mutant MCM2. This suppressor mutation was located in the HBO1 zinc finger. Taken together, these findings strongly suggest that the interaction between MCM2 and HBO1 is direct and mediated by the C2HC zinc finger of HBO1. The biochemical and genetic interactions of MYST family protein HBO1 with two components of the replication apparatus, MCM2 and ORC1, suggest that HBO1-associated HAT activity may play a direct role in the process of DNA replication.

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.

Collaborative role of E2F transcriptional activity and G1 cyclindependent kinase activity in the induction of S phase.

A considerable body of evidence points to a role for both cyclin E/cyclin-dependent kinase (cdk)2 activity and E2F transcription activity in the induction of S phase. We show that overexpression of cyclin E/cdk2 in quiescent cells induces S phase, that this coincides with an induction of E2F activity, and that coexpression of E2F enhances the cyclin E/cdk2-mediated induction of S phase. Likewise, E2F overexpression can induce S phase and does so in the apparent absence of cyclin E/cdk2 activity. In addition, although the inhibition of cyclin E/cdk2 activity blocks the induction of S phase after growth stimulation of normal mouse embryo fibroblasts, inhibition of cyclin E/cdk2 does not block S phase induction in Rb-/- cells where E2F activity is deregulated. These results point to the important roles for E2F and cyclin E/cdk2 in the induction of S phase. Moreover, the nature of the E2F targets and the suspected targets for cyclin E/cdk2 suggests a potential molecular mechanism for the collaborative action of cyclin E/cdk2 and E2F in the induction of S phase.

Differential regulation of transcription: repression by unactivated mitogen-activated protein kinase Kss1 requires the Dig1 and Dig2 proteins.

Kss1, a yeast mitogen-activated protein kinase (MAPK), in its unphosphorylated (unactivated) state binds directly to and represses Ste12, a transcription factor necessary for expression of genes whose promoters contain filamentous response elements (FREs) and genes whose promoters contain pheromone response elements (PREs). Herein we show that two nuclear proteins, Dig1 and Dig2, are required cofactors in Kss1-imposed repression. Dig1 and Dig2 cooperate with Kss1 to repress Ste12 action at FREs and regulate invasive growth in a naturally invasive strain. Kss1-imposed Dig-dependent repression of Ste12 also occurs at PREs. However, maintenance of repression at PREs is more dependent on Dig1 and/or Dig2 and less dependent on Kss1 than repression at FREs. In addition, derepression at PREs is more dependent on MAPK-mediated phosphorylation than is derepression at FREs. Differential utilization of two types of MAPK-mediated regulation (binding-imposed repression and phosphorylation-dependent activation), in combination with distinct Ste12-containing complexes, contributes to the mechanisms by which separate extracellular stimuli that use the same MAPK cascade can elicit two different transcriptional responses.

Repression of yeast Ste12 transcription factor by direct binding of unphosphorylated Kss1 MAPK and its regulation by the Ste7 MEK.

The mitogen-activated protein kinase (MAPK) Kss1 has a dual role in regulating filamentous (invasive) growth of the yeast Saccharomyces cerevisiae. The stimulatory function of Kss1 requires both its catalytic activity and its activation by the MAPK/ERK kinase (MEK) Ste7; in contrast, the inhibitory function of Kss1 requires neither. This study examines the mechanism by which Kss1 inhibits invasive growth, and how Ste7 action overcomes this inhibition. We found that unphosphorylated Kss1 binds directly to the transcription factor Ste12, that this binding is necessary for Kss1-mediated repression of Ste12, and that Ste7-mediated phosphorylation of Kss1 weakens Kss1-Ste12 interaction and relieves Kss1-mediated repression. Relative to Kss1, the MAPK Fus3 binds less strongly to Ste12 and is correspondingly a weaker inhibitor of invasive growth. Analysis of Kss1 mutants indicated that the activation loop of Kss1 controls binding to Ste12. Potent repression of a transcription factor by its physical interaction with the unactivated isoform of a protein kinase, and relief of this repression by activation of the kinase, is a novel mechanism for signal-dependent regulation of gene expression.

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.

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.