Diurnal variations of progesterone,testosterone, and androsta-1,4-diene-3,17-dione in the rumen and in vitro progesterone transformation by mixed rumen microorganisms of lactating dairy cows

Five Holstein lactating dairy cows fed 5 total mixed rations (TMR) with different forage combinations were used in a 5 × 5 Latin square design to investigate diurnal variations of progesterone (P4), testosterone, and androsta-1,4-diene-3,17-dione (ADD) concentrations in the rumen. Meanwhile, different P4 inclusion levels [0 (control), 2, 20, 40, 80, and 100 ng/mL in culture fluids] were incubated in vitro for 6, 12, 24, 36, 48, and 72 h together with rumen mixed microorganisms grown on a maize-rich feed mixture (maize meal:Chinese ryegrass hay = :1) with an aim to determine microbial P4 transformation into testosterone and ADD. Ruminal P4, testosterone, and ADD concentrations of lactating dairy cows were greater in the TMR with forage combination of corn silage plus alfalfa hay or Chinese wild ryegrass hay than the TMR with the corn stover-based forage combination. The diurnal fluctuation pattern showed that P4, testosterone, and ADD concentrations in the rumen were greater at nighttime than daytime and peaked 6 h after feeding in the morning or afternoon. The in vitro batch cultures showed that the P4 elimination rate was highest at the P4 addition of 20 ng/mL and declined with the further increased addition of P4. The treatments after dosing P4 exhibited a shorter time than the control group until half of the initial P4 inclusion was eliminated (i.e., half time), and the lowest half time (1.46 h) occurred at the P4 addition of 20 ng/mL. In summary, the ruminal steroids concentration was affected by forage type and quality, and the rumen microorganisms exhibited great ability to transform P4 into testosterone and ADD, depending on incubation time and initial P4 addition level, suggesting that the host might affect the metabolism of its rumen microorganisms via the endogenous steroids.     

Genetic analyses of claw health in Norwegian Red cows

The aim of this study was genetic analyses of claw health in Norwegian Red. Claw health status at claw trimming has, since 2004, been recorded in the Norwegian Dairy Herd Recording System. The claw trimmer records whether the cow has normal (healthy) claws or if one or more claw disorders are present. Nine defined claw disorders were recorded: corkscrew claw (CSC), heel horn erosion (HH), dermatitis (DE), sole ulcer (SU), white line disorder (WLD), hemorrhage of sole and white line (HSW), interdigital phlegmon (IDP), lameness (LAME), and acute trauma (AT). Data from 2004 to 2011, with a total of 204,892 claw health records, were analyzed. The disorders were defined as binary traits with 1 record per cow per lactation. Further, 3 groups of claw disorders were analyzed: infectious claw disorders (INFEC, containing HH, DE, and IDP); laminitis-related claw disorders (LAMIN, containing SU, WLD, and HSW); and overall claw disorder. The 9 single traits and the 3 groups were analyzed using univariate threshold sire models. Multivariate threshold models were performed for the 5 most frequent single traits (CSC, HH, DE, SU, and WLD) and for CSC together with the grouped traits INFEC and LAMIN. Posterior mean of heritability of liability ranged from 0.04 to 0.23, where CSC had the highest heritability. The posterior standard deviations of heritability were low, between 0.01 and 0.03, except for IDP (0.06). Heritability of liability to INFEC and LAMIN were both 0.11 and for overall claw disorders, the heritability was 0.13. Posterior means of the genetic correlation among the 5 claw disorders varied between 0.02 and 0.79, and the genetic correlations between DE and HH (0.65) and between WLD and SU (0.79) were highest. Genetic correlation between INFEC and CSC was close to zero (0.06), between LAMIN and CSC it was 0.31, and between LAMIN and INFEC it was 0.24. The results show that claw disorders are sufficiently heritable for genetic evaluation and inclusion in the breeding scheme. At present, data are scarce with few recorded daughters per sire. Claw trimming records from more herds would therefore be beneficial for routine genetic evaluation of claw health.     

Concentrations of progesterone in milk of cows administered an intravaginal progesterone insert

Milk from pregnant cows contains concentrations of progesterone (P4) considered safe for human consumption. The objective of this study was to determine if concentrations of P4 in milk during administration of an intravaginal progesterone insert (CIDR insert) are less than concentrations of P4 in milk associated with pregnancy. Results have implications for human use of milk from cows receiving CIDR inserts. Holstein cows (N = 64; > 40 and < 150 d after calving) were administered 25 mg of PGF2alpha i.m. (study d 0) and 20 cows detected in estrus from 2 to 4 d later were assigned randomly to either control (N = 10; no further treatment) or CIDR insert (N = 10; 1.38 g of P4) inserted on study d 17 (14 +/- 1 d after estrus) and removed 7 d later. Composite milk samples were collected contemporaneously from each of the 20 estrous cycling cows and from 10 pregnant cows (> or = 60 and < or = 220 d of gestation) twice daily from study d 17 to 27. Concentrations of P4 in defatted milk samples were quantified using a validated radioimmunoassay. Mean logs of areas under the curve of concentrations of P4 from the afternoon on study d 17 through the afternoon on study d 27 were 3.05 ng day/ml for control, 3.33 ng day/ml for CIDR insert, and 3.81 ng day/ml for pregnant cows. Therefore, increased P4 due to pregnancy was 0.76 ng day/ml (3.81-3.05), whereas the increase in P4 due to CIDR insert was only 0.28 ng day/ml (3.33-3.05). Applying a 95% confidence interval to 0.28 ng day/ml provided an upper value of 0.70 ng day/ml, lower than the increase due to pregnancy. Because milk from pregnant cows is considered safe for human consumption, it follows that milk from cows administered CIDR inserts should also be considered safe, based on concentrations of P4.     

Atypical progesterone profiles and fertility in Swedish dairy cows

The incidence of normal and atypical progesterone profiles in Swedish dairy cows was studied. Data were collected from an experimental herd over 15 yr, and included 1,049 postpartum periods from 183 Swedish Holstein and 326 Swedish Red and White dairy cows. Milk progesterone samples were taken twice weekly until initiation of cyclical ovarian activity and less frequently thereafter. Progesterone profiles were 1) normal profile: first rise in milk progesterone above the threshold value before d 56 postpartum, followed by regular cyclical ovarian activity (70.4%); 2) delayed onset of cyclical ovarian activity: low milk progesterone the first 56 d postpartum (15.6%); 3) cessation of cyclical ovarian activity: ovarian activity resumed within 56 d postpartum, but ceased for a period of 14 d or more (6.6%); and 4) prolonged luteal phase: ovarian activity resumed within 56 d postpartum, but milk progesterone remained elevated in the nonpregnant cow for a period of 20 d or more (7.3%). Swedish Holsteins had 1.5 times higher risk of atypical profile than Swedish Red and Whites. Risk of atypical profiles was 0.5 and 0.7 times lower for older cows compared with first-parity cows; 2.3 times higher for cows in tie-stalls compared with those in loose housing; 2.6 times higher for cows calving during winter compared with summer; 0.5 times lower for cows in earlier (1994-1999) calving-year groups compared with the most recent (2000-2002); 2.5 times higher for cows with planned extended calving interval compared with conventional calving interval; and 2.2 times higher for an atypical profile in previous lactation compared with a normal profile. Cows with atypical profiles had a 15-d increase in interval from calving to first artificial insemination and an 18-d increase in interval from calving to conception. Progesterone samples taken within the first 60 d postpartum were used to calculate the percentage of samples above the threshold value of luteal activity. This measure had a significantly different mean in profiles and can be used to separate delayed onset of cyclical ovarian activity profiles and prolonged luteal phase profiles from normal. Thereby, it may be a more effective tool than measurements based only on the onset of ovarian cyclical activity in genetic evaluation of early postpartum fertility in dairy cows.     

Ultrasonographical examination of bovine claws through the sole horn on weight-bearing claws

Claw horn disruptions in the bovine claw are believed to be a consequence of pressure on the sole corium from the third phalanx, which may be caused by a weakening of the suspensory apparatus in the claw. We aimed to develop an ultrasonographic method that would make it possible to measure the thickness of the soft tissue between the third phalanx and the sole horn on a weight-bearing claw. A device was developed to record the sole horn and soft tissue thickness indirectly through a polyethylene plate, and 52 feet from slaughtered cows were examined using ultrasonography, both directly and indirectly. Soft tissue and sole horn thickness measurements in the apex and the plantar part of the sole were compared with anatomical measurements of transected claws. To assess the method on weight-bearing versus non-weight-bearing claws, we examined the hind claws of 10 live cattle without transection. We found a weak correlation between the soft tissue thickness measured by ultrasound and anatomical measurements. A strong correlation was observed between the direct ultrasound approach and the developed indirect method. There was a considerable difference between weight-bearing and non-weight-bearing claws, signifying a weak or nonexistent correlation. However, this part of the study was only done on 10 live cows and the results should be interpreted with caution. We concluded that it would be possible to measure the soft tissue using an indirect ultrasound approach on a weight-bearing-claw standing on a polyethylene plate. The major difference between the results of weight-bearing versus non-weight-bearing claws suggests that future studies of the suspensory apparatus could focus on weight-bearing claws.     

Reproduction in dairy cows following progesterone insert presynchronization and resynchronization protocols

The objectives of this study were to evaluate the effects of an intravaginal insert containing progesterone (CIDR, controlled internal drug releasing) administered in presynchronization and resynchronization protocols on cyclicity, detection of estrus, pregnancy rate, and pregnancy loss to first AI; reinsemination patterns; and pregnancy rates to second postpartum AI before and after the time of first-service pregnancy diagnosis in dairy cows. Holstein cows (n = 1,052) were blocked by parity and BCS at 3 ± 3 d in milk (study d 0 = day of calving) and assigned randomly to 1 of 3 presynchronization treatments. During the presynchronization programs, all cows received 2 injections of PGF_2α, on study d 35 and 49. Cows enrolled in the control presynchronization treatment received AI after detected estrus from study d 49 to 62. Cows enrolled in the CIDR estrus-detection (CED) presynchronization treatment received a CIDR insert from study d 42 to 49 and AI on detection of estrus from d 49 to 62. Cows enrolled in the CIDR timed AI (CTAI) presynchronization treatment received the same treatment as CED, but were subjected to timed AI on study d 72 after the Ovsynch (GnRH, 7 d PGF_2α, 2 d GnRH, 24 h timed AI) protocol. The control and CED cows not inseminated by study d 62 were enrolled in the Ovsynch protocol on the same day and received timed AI on study d 72. After first AI, cows were assigned to no resynchronization (RCON) or resynchronization with a CIDR insert (RCIDR) between 14 and 21 d after AI. Blood samples collected on study d 35, 49, and 62 were analyzed for concentrations of progesterone and cows were classified as anestrous when progesterone was <1.0 ng/mL in the first 2 samples. On study d 62, anestrous cows with progesterone ≥ 1.0 ng/mL were classified as having resumed cyclicity. Pregnancy was diagnosed at 31 and 60 d after first AI and at 42 d after second AI. A greater proportion of anestrous cows in CED and CTAI became cyclic by d 62 postpartum than control cows. Resynchronization with the CIDR insert increased the pregnancy rate at 31 d after first AI in CED and CTAI, and at 60 d after AI in all cows because of reduced pregnancy loss. These results indicate that presynchronization with the CIDR insert increased induction of cyclicity in anestrous cows and that resynchronization with the CIDR insert did not affect the reinsemination rate but did reduce pregnancy loss and increased the pregnancy rate at 60 d after first AI.     

Genetic correlations between claw health and feet and leg conformation in Norwegian Red cows

The aim of this study was to estimate genetic correlations between claw disorders and feet and leg conformation traits in Norwegian Red cows. A total of 188,928 cows with claw health status recorded at claw trimming from 2004 to September 2013 and 210,789 first-lactation cows with feet and leg conformation scores from 2001 to September 2013 were included in the analyses. Traits describing claw health were corkscrew claw, infectious claw disorders (dermatitis, heel horn erosion, and interdigital phlegmon), and laminitis-related claw disorders (sole ulcer, white line disorder, and hemorrhage of sole and white line). The feet and leg conformation traits were rear leg rear view (new and old definition), rear leg side view, foot angle, and hoof quality. Feet and leg conformation traits were scored linearly from 1 to 9, with optimum scores depending on the trait. Claw disorders were defined as binary (0/1) traits for each lactation. Threshold sire models were used to model claw disorders, whereas the feet and leg conformation traits were described by linear sire models. Three multivariate analyses were performed, each including the 5 feet and leg conformation traits and 1 of the 3 claw disorders at a time. Posterior means of heritability of liability of claw disorders ranged from 0.10 to 0.20 and heritabilities of feet and leg conformation traits ranged from 0.04 to 0.11. Posterior standard deviation of heritability was ≤0.01 for all traits. Genetic correlations between claw disorders and feet and leg conformation traits were all low or moderate, except between corkscrew claw and hoof quality (−0.86), which are supposed to measure the same trait. The genetic correlations between rear leg rear view (new) and infectious claw disorders (−0.20) and laminitis-related claw disorders (0.26), and between hoof quality and laminitis-related claw disorders (−0.33) were moderate. Eight of the 15 genetic correlations between claw disorders and feet and leg conformation traits had 0 included in the 95% highest posterior density interval. These results imply that selection for feet and leg conformation is not an efficient approach to genetically improve claw health in Norwegian Red cattle.     

A multistate modeling approach to investigate long-term effects of claw horn disruption lesions and early lesion development in dairy cows

Claw horn disruption lesions (CHDL) are a leading cause of lameness in dairy cattle, and the development, effect, and pathology of these lesions remains an open area of interest within dairy cattle health. Current literature typically attempts to measure the effect of risk factors on the development of CHDL over a relatively short time period. Further understanding of the interaction of CHDL and the long-term effect of early CHDL in a cow's life remains an important area of research which is so far mostly unexplored. In this study 57,974 cows from 1,332 herds were selected and their regular claw trimming records containing important claw health information were used to model the long-term effect of lesions in a cow's lifetime in a 6-state multistate model. A multistate model predicts the time before transition from any one state to another and the probability of transition to a future state. The 6 lesion states that were modeled were as follows: never had a lesion, first recorded lesion event, no recorded lesion after first lesion event, second or subsequent recorded lesion event, no recorded lesion after second or subsequent lesion event, and culled. The effect of various cow level covariates on the transition probabilities between various states was tested. For the first time, this study shows the importance and effect of the first lesion and other cow level factors on long-term claw health. Model results showed that the timing and severity of the first recorded lesion event significantly influenced the likelihood of a future lesion being present. Cows with CHDL present within the 180 d of first calving had a short-term increased risk and long-term decreased risk of a future lesion, compared with cows that present with CHDL later than 180 d of first lactation. Moreover, presence of a severe first lesion increased a cow's risk of a future lesion being present. The model was used to evaluate the relative difference between high-risk cows (age of first calving ≥793 d, breeding values in the lowest quartile) and low-risk cows (age of first calving ≤718 d, breeding values in the highest quartile). Our results indicated that these low-risk cows present with a lesion on an average 3 mo later than high-risk cows. Furthermore, results from the model evaluation of a simulated herd with cows with breeding values in the higher quartile indicated that cows present with a CHDL on an average 7.5 mo later compared with a herd where cows have breeding values distributed in a lower quartile.     

Effects of induced weight shift in the hind limbs on claw loads in dairy cows

The modern dairy industry is plagued by a high prevalence of claw horn lesions in cows, which cause lameness, affect well-being, limit milk production, and are responsible for premature removal of cows from the herd. The lateral hind claws are primarily affected, and this has been linked to a relatively higher load being exerted on the lateral claws when cows shift weight from one hind limb to the other. The vertical ground reaction forces and mean and maximum pressures under the claws were measured in 40 nonlame dairy cows before and during a shift in weight from one hind limb to the other, which was accomplished by applying pressure manually to one side of the pelvis. During square standing on firm ground, about two-thirds of the entire hind limb load was exerted on the 2 lateral claws, and the remaining one-third was exerted on the medial claws combined. At the moment of maximum weight shift, the lateral claw of the loaded limb bore almost two-thirds of the entire load of both hind limbs, with the heel zone bearing almost half of the load of both hind limbs. Subsequently, the load of the lateral claw of the contralateral hind limb decreased, as did the load of both medial claws. Thus, the weight redistribution had occurred predominantly between the lateral hind claws. The high load exerted on a lateral hind claw during weight shift and at maximum weight shift is assumed to play a role in the pathogenesis of claw horn lesions, particularly when accentuated by a softened claw horn and hard flooring.     

Concentration of progesterone during the development of the ovulatory follicle: II. Ovarian and uterine responses

Two experiments evaluated the influence of altering the concentrations of progesterone during the development of the ovulatory follicle on the composition of the follicular fluid, circulating LH and PGF_2α metabolite (PGFM), and expression of endometrial progesterone receptor and estrogen receptor-α. In both experiments, the estrous cycles were presynchronized (GnRH and progesterone insert followed by insert removal and PGF_2α 7 d later, and GnRH after 48 h) and cows were then enrolled in 1 of 2 treatments 7 d later (study d −16): high progesterone (HP) or low progesterone (LP). In experiment 1 (n = 19), cows had their estrous cycle synchronized starting on study d −9 (GnRH and progesterone insert on d −9, and insert removal and PGF_2α on d −2). In experiment 2 (n = 25), cows were submitted to the same synchronization protocol as in experiment 1, but had ovulation induced with GnRH on study d 0. In experiment 1, plasma was sampled on d −4 and analyzed for concentrations of LH; the dominant follicle was aspirated on d 0 and the fluid analyzed for concentrations of progesterone, estradiol, and free and total IGF-1. In experiment 2, follicular development and concentrations of progesterone and estradiol in plasma were evaluated until study d 16. Uterine biopsies were collected on d 12 and 16 for progesterone receptor and estrogen receptor-α protein abundance. An estradiol/oxytocin challenge for PGFM measurements in plasma was performed on d 16. In experiments 1 and 2, LP cows had lower plasma concentrations of progesterone and greater concentrations of estradiol, and had larger ovulatory follicle diameter (20.4 vs. 17.2 mm) at the end of the synchronization protocol than HP cows. Concentration of LH tended to be greater for LP than HP cows (0.98 vs. 0.84 ng/mL). The dominant follicle of LP cows had greater concentration of estradiol (387.5 vs. 330.9 ng/mL) and a lower concentration of total IGF-1 (40.9 vs. 51.7 ng/mL) than that of HP cows. In experiment 2, estradiol and progesterone concentrations did not differ between treatments from d 0 to 16; however, the proportion of cows with a short luteal phase tended to increase in LP than HP (25 vs. 0%). Concentrations of PGFM were greater for LP than HP. Uterine biopsies had a greater abundance of progesterone receptor, and tended to have less estrogen receptor-α abundance on d 12 compared with d 16. An interaction between treatment and day of collection was detected for estrogen receptor-α because of an earlier increase in protein abundance on d 12. Reduced concentrations of progesterone during the development of the ovulatory follicle altered follicular dynamics and follicular fluid composition, increased basal LH concentrations, and prematurely increased estrogen receptor-α abundance and exacerbated PGF_2α release in the subsequent estrous cycle.     

Characterization of progesterone profiles in fall-calving Norwegian Red cows

Progesterone profiles in Norwegian Red cows were categorized, and associations between the occurrence of irregularities in the profiles and the commencement of luteal activity were investigated. The cows were managed in 3 feeding trials from 1994 to 2001 and from 2005 to 2008 at the Norwegian University of Life Sciences. The cows were followed from calving, and the milk samples collected represented 502 lactations from 302 cows. Milk samples for progesterone analysis were taken 3 times weekly from 1994 throughout 1998 and from 2005 to 2008 and 2 times weekly from 1999 to 2001. Commencement of luteal activity was defined as the first day of 2 consecutive measurements of progesterone concentration ≥3 ng/mL not earlier than 10 d after calving. Delayed ovulation type I was defined as consistently low progesterone concentration, <3 ng/mL for ≥50 d postpartum. Delayed ovulation type II was defined as prolonged interluteal interval with milk progesterone measurements <3 ng/mL for ≥12 d between 2 luteal phases. Persistent corpus luteum (PCL) type I was defined as delayed luteolysis with milk progesterone ≥3 ng/mL for ≥19 d during the first estrous cycle postpartum. Persistent corpus luteum type II was defined as delayed luteolysis with milk progesterone ≥3 ng/mL for ≥19 d during subsequent estrous cycles before first artificial insemination. Delayed ovulation type I was present in 14.7%, delayed ovulation type II in 2.8%, PCL type I in 6.7%, and PCL type II in 3.3% of the profiles. Commencement of luteal activity was related to milk yield, parity, PCL type I, and the summated occurrence of PCL type I and II. The least squares means for the interval to commencement of luteal activity were 24.2 d when PCL type I and II were present and 29.5 d when PCL type I and II were absent. The likelihood of pregnancy to first service was not affected in cows with a history of PCL when artificial insemination was carried out at progesterone concentrations <3 ng/mL (i.e., during estrus); however, cows that had experienced PCL were more likely to be inseminated during a luteal phase. The occurrence of delayed ovulation and PCL in Norwegian Red cows was less than that reported in most other dairy populations.     

Concentration of progesterone during the development of the ovulatory follicle: I. Ovarian and embryonic responses

Objectives were to evaluate the effects of differing progesterone concentrations during follicle development on follicular dynamics, fertilization, and embryo quality. Lactating Holstein cows (n = 154) were assigned randomly to 1 of 2 treatments. Cows underwent a presynchronization of the estrous cycle composed of an injection of GnRH concurrently with the placement of a progesterone insert, an injection of PGF_2α and insert removal 7 d later, and a second injection of GnRH 48 h later (study d −16). All cows were then submitted to a hormonal protocol identical to the presynchronization program starting on d 7 of the estrous cycle (study d −9). Cows enrolled in the high progesterone (HP) treatment received no further treatment. Cows in the low progesterone (LP) treatment received additional PGF_2α injections on study d −14, −13.5, and −13 and again on study d −9, −7, −6.5, and −6. Ovaries were evaluated by ultrasonography, and blood was sampled for concentrations of progesterone and estradiol throughout the study. Uteri were flushed 6 d after artificial insemination (AI) and recovered oocytes-embryos were evaluated. Concentrations of progesterone were less for LP cows from study d −7 to −2; concentrations of estradiol at PGF_2α and at the last GnRH of synchronization were greater for LP than HP. The proportion of cows in estrus at AI was greater for LP than for HP (38.0 vs. 5.3%). Ovulatory follicles of LP cows had larger diameters at the injections of PGF_2α (17.2 vs. 14.6 mm) and final GnRH (19.4 vs. 16.9%) of the synchronization, which resulted in a larger diameter of the corpus luteum 6 d after AI (24.3 vs. 22.6 mm). Double ovulation after the last GnRH of the synchronization was increased in LP (18.6%) compared with HP (4.5%). Fertilization rate was similar and averaged 82.7%. The proportion of embryos and oocytes-embryos classified as grades 1 and 2, proportion of degenerated embryos, and unfertilized-degenerated oocytes-embryos were not different between LP and HP. Number of blastomeres did not differ between LP and HP, but the proportion of live blastomeres tended to be less for LP than HP (94.2 vs. 98.7%). Reducing progesterone concentrations during the synchronization program altered concentrations of estradiol and follicular dynamics, but resulted in similar fertilization and only minor changes in embryo quality.     

Short communication: Effects of the progesterone receptor variants on fertility traits in cattle

The progesterone receptor (PGR) gene is a key factor in the initiation and maintenance of pregnancy and in embryo development. Currently, it is unknown what variants of the PGR gene are related to fertility traits in cattle. Identification of such variants would allow the implementation of marker-assisted selection in breeding schemes. The objective of this study was to investigate the association of single nucleotide polymorphisms (SNP) of PGR with fertility traits in Holstein dairy cattle. An in vitro fertilization system was used to maximize the efficiency of the identification of genetic factors affecting fertility. This in vitro fertilization system would allow the assessment of fertilization and embryonic survival rates independently of influences from the uterine environment. A total of 5,566 fertilization attempts were performed, and a total of 3,679 embryos were produced using oocytes from 324 Holstein cows and semen from 10 Holstein bulls. Sequencing of pooled DNA samples from ovaries revealed an SNP (G/C) in intron 3 of PGR. A generalized linear model was used to analyze the association of this SNP with fertilization and embryonic survival rates for each ovary. Oocytes obtained from CC ovaries showed a 61% fertilization rate, compared with 68 and 69% for GC and GG ovaries, respectively. The survival rate of embryos produced from GG ovaries was 5 and 6% higher than that of GC and CC ovaries. These results indicate that the PGR SNP could be used in marker-assisted selection breeding programs in Holstein dairy cattle.     

Effects of routine treatment with nonsteroidal anti-inflammatory drugs at calving and when lame on the future probability of lameness and culling in dairy cows: A randomized controlled trial

Claw horn lesions (CHL) are reported as the most common cause of lameness in intensive dairy systems. Despite their prevalence, the underlying pathological mechanisms and preventive strategies for CHL remain poorly understood. Recent advances have pointed to the role of inflammation in disease aetiopathogenesis. Moderating inflammation from first calving may lead to long-term benefits and a viable intervention for treating and preventing disease. We conducted a 34-mo randomized controlled trial to investigate the effects of routine treatment with the nonsteroidal anti-inflammatory drug ketoprofen at calving and during treatment for lameness, on the future probability of lameness and culling, caused by exposure to normal farm conditions. A cohort of dairy heifers were recruited from a single, commercial dairy herd between January 8, 2018, and June 22, 2020, and randomly allocated to one of 4 treatment groups before first calving. The lactating herd was lameness scored every 2 wk on a 0 to 3 scale, to identify animals that became lame (single score ≥2a) and hence required treatment. Animals in group 1 received a therapeutic trim and a hoof block on the sound claw (if deemed necessary) every time they were treated for lameness. Animals in group 2 received the same treatment as group 1 with the addition of a 3-d course of ketoprofen (single dose daily) every time they were treated for lameness. Animals in group 3 received the same treatment as group 2 with the addition of a 3-d course of ketoprofen (single dose daily) starting 24 to 36 h after each calving. Animals in group 4 received a 3-d course of ketoprofen (single dose daily) every time they were identified with lameness. No therapeutic trim was administered to this group, unless they were identified as severely lame (a single score ≥3a). Animals were followed for the duration of the study (ending October 23, 2020). Probability of lameness was assessed by a lameness outcome score collected every 14 d. Data on culling was extracted from farm records. One hundred thirty-two animals were recruited to each group, with data from 438 animals included in the final analysis (111 in group 1, 117 in group 2, 100 in group 3, and 110 in group 4). Mixed effect logistic regression models were used to evaluate the effect of treatment group on the ongoing probability of lameness. Compared with the control group (group 1), animals in group 3 were less likely to become lame (odds ratio: 0.66) and severely lame (odds ratio: 0.28). A Cox proportional hazards survival model was used to investigate the effect of treatment group on time to culling. Compared with group 1, animals in groups 2 and 3 were at reduced risk of culling (hazard ratios: 0.55 and 0.56, respectively). The lameness effect size we identified was large and indicated that treating a cohort of animals with the group 3 protocol, would lead to an absolute reduction in population lameness prevalence of approximately 10% and severe lameness prevalence of 3%, compared with animals treated in accordance with conventional best practice (group 1).     

Pregnancy in dairy cows after synchronized ovulation regimens with or without presynchronization and progesterone

Two experiments examined pregnancy after synchronized ovulation (Ovsynch) with or without progesterone (P4) administered via controlled internal drug release (CIDR) intravaginal inserts. In experiment 1, 262 lactating cows in one herd were in 3 treatments: Ovsynch (n = 91), Ovsynch + CIDR (n = 91), and control (n = 80). The Ovsynch protocol included injections of GnRH 7 d before and 48 h after an injection of PGF20. Timed artificial insemination (TAI; 57 to 77 d postpartum) was 16 to 20 h after the second GnRH injection. Cows in the Ovsynch + CIDR group also received a CIDR (1.9 g of P4) insert for 7 d starting at first GnRH injection. Control cows received A-I when estrus was detected using an electronic estrus detection system. Based on serum P4, 44.1% of cows were cyclic before Ovsynch. Pregnancy rates at 29 d (59.3 vs. 36.3%) and 57 d (45.1 vs. 19.8%) after TAI and embryo survival (75.9 vs. 54.5%) from 29 to 57 d were greater for Ovsynch + CIDR than for Ovsynch alone. In experiment 2, 630 cows in 2 herds received TAI at 59 to 79 d postpartum after 6 treatments. Estrous cycles were either presynchronized (2 injections of PGF2alpha 14 d apart; n = 318) or not presynchronized (n = 312). Within those groups, Ovsynch was initiated 12 d after second presynchronization PGF2alpha, and used alone (n = 318) or with CIDR inserts for 7 d (1.38 g of P4/insert, n = 124 or 1.9 g of P4/insert, n = 188). Before Ovsynch, 80% of cows were cyclic. Presynchronization increased pregnancy (46.8 vs. 37.5%) at 29 d after TAI, but CIDR inserts had no effect on pregnancy in experiment 2. Overall embryonic survival between 29 and 57 d in experiment 2 was 57.7%. Use of CIDR inserts with Ovsynch improved conception and embryo survival in experiment 1 but not in experiment 2, in part due to differing proportions of cyclic cows at the outset. Presynchronization before Ovsynch enhanced pregnancy rate.     

Effects of supplemental progesterone after artificial insemination on expression of interferon-stimulated genes and fertility in dairy cows

The objectives of the current study were to evaluate the effects of supplemental progesterone after artificial insemination (AI) on expression of IFN-stimulated genes (ISG) in blood leukocytes and fertility in lactating dairy cows. Weekly cohorts of Holstein cows were blocked by parity (575 primiparous and 923 multiparous) and method of insemination (timed AI or AI on estrus) and allocated randomly within each block to untreated controls, a controlled internal drug release (CIDR) containing 1.38 g of progesterone from d 4 to 18 after AI (CIDR4), or a CIDR on d 4 and another on d 7 after AI and both removed on d 18 (CIDR4+7). Blood was sampled to quantify progesterone concentrations in plasma and mRNA expression in leukocytes for the ubiquitin-like IFN-stimulated gene 15-kDa protein (ISG15) and receptor transporter protein-4 (RTP4) genes. Pregnancy was diagnosed on d 34 ± 3 and 62 ± 3 after AI. Treatment increased progesterone concentrations between d 5 and 18 after AI in a dose-dependent manner (control = 3.42, CIDR4 = 4.97, and CIDR4+7 = 5.46 ng/mL). Cows supplemented with progesterone tended to have increased luteolysis by d 19 after AI (control = 17.2; CIDR4 = 29.1; CIDR4+7 = 30.2%), which resulted in a shorter AI interval for those reinseminated after study d 18. Pregnancy upregulated expression of ISG in leukocytes on d 19 of gestation, but supplementing progesterone did not increase mRNA abundance for ISG15 and RTP4 on d 16 after insemination and tended to reduce mRNA expression on d 19 after AI. For RTP4 on d 19, the negative effect of supplemental progesterone was observed only in the nonpregnant cows. No overall effect of treatment was observed on pregnancy per AI on d 62 after insemination and averaged 28.6, 32.7, and 29.5% for control, CIDR4, and CIDR4+7, respectively. Interestingly, an interaction between level of supplemental progesterone and method of AI was observed for pregnancy per AI. For cows receiving exogenous progesterone, the lower supplementation with CIDR4 increased pregnancy per AI on d 62 in cows inseminated following timed AI (CIDR4 = 39.2; CIDR4+7 = 27.5%); in those inseminated following detection of estrus, however, the use of a second insert on d 7 resulted in greater pregnancy per AI (CIDR4 = 26.9; CIDR4+7 = 31.5%). Pregnancy loss did not differ among treatments. Supplemental progesterone post-AI using a single intravaginal insert on d 4 was beneficial to pregnancy in cows inseminated following timed AI, but incremental progesterone with a second insert on d 7 did not improve fertility of dairy cows.     

A prospective cohort study of digital cushion and corium thickness. Part 2: Does thinning of the digital cushion and corium lead to lameness and claw horn disruption lesions?

The aim of this study was to determine whether a decrease in thickness of the sole soft tissues (SST) beneath the flexor tuberosity of the distal phalanx (i.e., the digital cushion and corium) predisposed a claw to develop claw horn disruption lesions (CHDL) or a leg to lameness. Data were analyzed from a longitudinal study of 179 cows, which had been examined at 5 assessment points ?8, +1, +9, +17, and +29 wk relative to their first, second, third, or fourth calving. At each assessment point, SST were measured using ultrasonography. Additional assessment point data included sole lesions and back fat thickness (BFT), and cows had been locomotion scored every 2 wk from calving. One hundred fifty-eight cows completed the study. Separate logistic regression survival analyses were constructed to assess the outcomes, either lameness on a leg or CHDL on a claw; combinations of lameness and lesions were tested as outcomes. Cow level variables tested included farm and lactation number. Variables were tested describing previous SST thickness, minimum previous SST thickness, BFT, and change in either variable between prior assessment points. Prior lesions/lameness strongly predicted repeat cases and the final models had the outcome first lesion or lameness on a claw or leg. In the reported lameness models, lameness was defined as a leg being recorded as lame twice within 3 consecutive scores, and in the reported lesion models, lesion was defined as the first presence of either a sole ulcer or a severe sole hemorrhage on a claw. Thin SST increased the likelihood of lesion occurrence; thin SST on the lateral claw predicted subsequent lameness on a leg. Thin BFT and thinning of BFT between previous assessment points increased the likelihood of future lesion occurrence. Thin SST and thinning of BFT had additional effects on the likelihood of lesion occurrence, suggesting that BFT and sole SST had independent effects on lesion occurrence. However, change in SST thickness between assessment points did not influence the likelihood of future lesions or lameness. This suggests that thin SST were not simply a result of depletion of body fat and challenges the theory that thinning of the digital cushion with body fat mobilization leads to CHDL. Other possible mechanisms by which SST become thin are discussed and could include changes in integrity of the suspensory apparatus with physiological events.     

Genetic analysis of atypical progesterone profiles in Holstein-Friesian cows from experimental research herds

The objective of this study was to quantify the genetic variation in normal and atypical progesterone profiles and investigate if this information could be useful in an improved genetic evaluation for fertility for dairy cows. The phenotypes derived from normal profiles included cycle length traits, including commencement of luteal activity (C-LA), interluteal interval, luteal phase length. and interovulatory interval. In total, 44,977 progesterone test-day records were available from 1,612 lactations on 1,122 primiparous and multiparous Holstein-Friesian cows from Ireland, the Netherlands, Sweden, and the United Kingdom. The atypical progesterone profiles studied were delayed cyclicity, prolonged luteal phase, and cessation of cyclicity. Variance components for the atypical progesterone profiles were estimated using a sire linear mixed model, whereas an animal linear mixed model was used to estimate variance components for the cycle length traits. Heritability was moderate for delayed cyclicity (0.24 ± 0.05) and C-LA (0.18 ± 0.04) but low for prolonged luteal phase (0.02 ± 0.04), luteal phase length (0.08 ± 0.05), interluteal interval (0.08 ± 0.14), and interovulatory interval (0.03 ± 0.04). No genetic variation was detected for cessation of cyclicity. Commencement of luteal activity, luteal phase length, and interovulatory interval were moderately to strongly genetically correlated with days from calving to first service (0.35 ± 0.12, 0.25 ± 0.14, and 0.76 ± 0.24, respectively). Delayed cyclicity and C-LA are traits that can be important in both genetic evaluations and management of fertility to detect (earlier) cows at risk of compromised fertility. Delayed cyclicity and C-LA were both strongly genetically correlated with milk yield in early lactation (0.57 ± 0.14 and 0.45 ± 0.09, respectively), which may imply deterioration in these traits with selection for greater milk yield without cognizance of other traits.     

Resynchronization strategies to improve fertility in lactating dairy cows utilizing a presynchronization injection of GnRH or supplemental progesterone: I. Pregnancy rates and ovarian responses

Objectives were to evaluate 3 resynchronization protocols for lactating dairy cows. At 32 ± 3 d after pre-enrollment artificial insemination (AI; study d −7), 1 wk before pregnancy diagnosis, cows from 2 farms were enrolled and randomly assigned to 1 of 3 resynchronization protocols after balancing for parity, days in milk, and number of previous AI. All cows were examined for pregnancy at 39 ± 3 d after pre-enrollment AI (study d 0). Cows enrolled as controls (n = 386) diagnosed not pregnant were submitted to a resynchronization protocol (d 0-GnRH, d 7-PGF_2α, and d 10-GnRH and AI) on the same day. Cows enrolled in the GGPG (GnRH-GnRH-PGF_2α-GnRH) treatment (n = 357) received a GnRH injection at enrollment (d −7) and if diagnosed not pregnant were submitted to the resynchronization protocol for control cows on d 0. Cows enrolled in CIDR treatment (n = 316) diagnosed not pregnant received the resynchronization protocol described for control cows with addition of a controlled internal drug release (CIDR) insert containing progesterone (P₄) from d 0 to 7. In a subgroup of cows, ovaries were scanned and blood was sampled for P₄ concentration on d 0 and 7. After resynchronized AI, cows were diagnosed for pregnancy at 39 ± 3 and 67 ± 3 d (California herds) or 120 ± 3 d (Arizona herds). Cows in the GGPG treatment had more corpora lutea than CIDR and control cows on d 0 (1.30 ± 0.11, 1.05 ± 0.11, and 1.05 ± 0.11, respectively) and d 7 (1.41 ± 0.14, 0.97 ± 0.13, and 1.03 ± 0.14, respectively). A greater percentage of GGPG cows ovulated to GnRH given on d 0 compared with CIDR and control cows (48.4, 29.6, and 36.6%, respectively), but CIDR and control did not differ. At 39 ± 3 d after resynchronized AI, pregnancy per AI (P/AI) was increased in GGPG (33.6%) and CIDR (31.3%) cows compared with control (24.6%) cows. At 67 or 120 ± 3 d after resynchronized AI, P/AI of GGPG and CIDR cows was increased compared with control cows (31.2, 29.5, and 22.1%, respectively). Presynchronizing the estrous cycle of lactating dairy cows with a GnRH 7 d before the start of the resynchronization protocol or use of a CIDR insert within the resynchronization protocol resulted in greater P/AI after resynchronized AI compared with control cows.     

Estimating probability of insemination success using milk progesterone measurements

The aim of this study was to quantify the effects of progesterone profile features and other cow-level factors on insemination success to provide a real-time predictor equation of probability of insemination success. Progesterone profiles from 26 dairy herds were analyzed and the effects of profile features (progesterone slope, cycle length, and cycle height) and cow traits (milk yield, parity, insemination during the previous estrus) on likelihood of artificial insemination success were estimated. The equation was fitted on a training data set containing data from 16 herds (6,246 estrous cycles from 3,404 lactations). The equation was tested on a testing data set containing data from 10 herds (8,105 estrous cycles from 3,038 lactations). Predictors were selected to be implemented in the final equation if adding them to a base model correcting for timing of insemination and parity decreased the overall likelihood distance of the model. Selected variables (cycle length, milk yield, cycle height, and insemination during the previous estrus) were used to build the final model using a stepwise approach. Predictors were added 1 by 1 in different order, and the model that had the smallest likelihood distance was selected. The final equation included the variables timing of insemination, parity, milk yield, cycle length, cycle height, and insemination during the previous estrus, respectively. The final model was applied to the testing data set and area under the curve (AUC) was calculated. On the testing data set, the final model had an AUC of 58%. When the farm effect was taken into account, the AUC increased to 63%. This equation can be implemented on farms that monitor progesterone and can support the farmer in deciding when to inseminate a cow. This can be the first step in moving the focus away from the current paradigm associated with poorer estrus detection, where each detected estrus is automatically inseminated, to near perfect estrus detection, where the question is which estrous cycle is worth inseminating?