The objective was to determine if using a Double-Ovsynch protocol [DO; Pre-Resynch: GnRH-7 d-PGF(2α)-3 d-GnRH, 7 d later Breeding-Resynch: GnRH-7 d-PGF(2α)-56 h-GnRH-16 h-timed artificial insemination (TAI)] to resynchronize ovulation after a previous TAI would increase synchrony and pregnancies per AI (P/AI) compared with an Ovsynch protocol initiated 32 d after TAI (D32; GnRH-7 d-PGF(2α)-56 h-GnRH-16 h-TAI). Lactating Holstein cows at various days in milk and prior AI services were blocked by parity and randomly assigned to resynchronization treatments. All DO cows received the first GnRH injection of Pre-Resynch 22 d after TAI, and cows (n=981) diagnosed not pregnant using transrectal ultrasonography 29 d after TAI continued the protocol. Pregnancy status for all D32 cows was evaluated 29 d after TAI so fertility and pregnancy loss could be compared with that of DO cows. All D32 cows received the first GnRH injection of Ovsynch 32 d after TAI, and cows (n=956) diagnosed not pregnant using transrectal palpation 39 d after TAI continued the protocol. In a subgroup of cows from each treatment, ultrasonography (n=751) and serum progesterone (P4) concentrations (n=743) were used to determine the presence of a functional corpus luteum (CL) and ovulation to the first GnRH injection of D32 and Breeding-Resynch of DO (GnRH1), luteal regression after PGF before TAI, and ovulation to the GnRH injection before TAI (GnRH2). Overall, P/AI 29 d after TAI was not affected by parity and was greater for DO compared with D32 cows (39 vs. 30%). Pregnancy loss from 29 to 74 d after TAI was not affected by parity or treatment. The percentage of cows with a functional CL (P4 ≥1.0 ng/mL) at GnRH1 was greater for DO than D32 cows (81 vs. 58%), with most DO cows having medium P4 (60%; 1.0 to 3.49 ng/ml), whereas most D32 cows had either low (42%; <1.0 ng/mL) or high (36%; ≥3.5 ng/mL) P4 at GnRH1. Ovulation to GnRH1 was similar between treatments but was affected by serum P4 at GnRH. Cows with low P4 (<1.0 ng/mL) had the greatest ovulatory response (59%), followed by cows with medium (≥1.0 to 3.49 ng/mL; 38%) and then high (≥3.50 ng/mL; 16%) P4 at GnRH1. A greater percentage of DO cows were synchronized compared with D32 cows (72 vs. 51%) primarily due to a greater percentage of D32 than DO cows without a functional CL at the PGF injection before TAI (35 vs. 17%) or without complete CL regression before GnRH2 (17 vs. 7%). We conclude that DO increased fertility of lactating dairy cows during a resynchronization program primarily by increasing synchronization of cows during the Ovsynch protocol before TAI.

The objectives of this observational study were (1) to assess the time from the appearance of the amniotic sac (AS) or feet outside the vulva to birth in Holstein cows (primiparous and multiparous) with (dystocia) or without assistance (eutocia) at calving, and (2) to estimate reference times to be used as guidelines for obstetric intervention in Holstein cows that need assistance during difficult births. Cows (n=92) from 1 commercial dairy operation were used in this study. Periparturient dairy cows (primiparous, n=58; multiparous, n=34) were placed in a maternity pen and constantly monitored until birth. The calving ease of cows, time from AS or feet appearance to birth, calving progress from a subset of 15 cows (frequency and duration of abdominal contractions during labor), calf birth weight, calf sex, and stillbirths (born dead or died within 24h after birth) were recorded. The reference times for obstetric intervention during dystocia were estimated based on values from unassisted births (normal). The normal range of times from the appearance of AS or feet outside the vulva to birth was estimated based on the mean+2 standard deviations (SD) of unassisted births. According to farm protocol, assistance was provided to cows without calving progress 80 min after AS appearance or earlier (e.g., to correct malpositions). Cows with dystocic births had a longer time from AS appearance to birth and increased incidence of stillbirth compared with cows with eutocic calvings. After the appearance of the AS, calving progress was evident every 15 min for eutocic births. The estimated reference times (mean+2 SD) from AS appearance to birth were 69.7 min and from feet appearance to birth were 64.6 min for eutocic births. Findings from this study suggested that calving personnel should start assisting cows 70 min after AS appearance (or 65 min after feet appearance) outside the vulva. The time spent in labor (straining) combined with the time from the appearance of the AS or feet to birth, and the assessment of calving progress (as described for eutocic births) should be used as guidelines for obstetric intervention during difficult births under field conditions. These reference times should be interpreted in combination with adequate obstetrical knowledge and examination.

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The objectives of the present case-control study were to assess (1) daily activity patterns (lying time, number of steps, number of lying bouts, and lying bout duration), and (2) circulating concentrations of biomarkers of pain (substance P), inflammation (haptoglobin), and stress (cortisol) in lactating dairy cows diagnosed with clinical metritis. Lactating dairy cows (n = 200) from 2 commercial dairy herds were enrolled in the present study. Cows diagnosed with clinical metritis (n = 100) at 7 ± 3 d in milk were matched according to lactation and days in milk to cows without clinical metritis (NO-CM; n = 100). On study d 1, clinical metritis was diagnosed (using a Metricheck device, Simcro Tech Ltd., Hamilton, New Zealand) by the presence of watery, reddish, or brownish foul-smelling vaginal discharge, and blood samples were collected for assessment of circulating concentration of substance P, haptoglobin, cortisol, total calcium, β-hydroxybutyrate, and blood cells. In addition, on study d 1 body condition of cows was visually assessed, and activity monitors were placed on the hind leg of a subset of cows (CM, n = 56; CON, n = 56) and were kept until study d 7. Cows showing any other signs of other diseases were not included in the study. Cows with clinical metritis tended to spend more time lying (CM = 628.92 min/d; NO-CM = 591.23 min/d) compared with NO-CM cows. Activity analysis by parity revealed that primiparous cows with clinical metritis spent more time lying compared with primiparous cows without clinical metritis. However, no differences in daily lying time were observed between multiparous cows with and without clinical metritis. Furthermore, cows in the CM group had a higher circulating concentration of substance P (CM = 47.15 pg/mL; NO-CM = 37.73 pg/mL) and haptoglobin (CM = 233.00 µg/mL; NO-CM = 99.98 µg/mL) when compared with NO-CM cows. Cows with clinical metritis had lower body condition score, and a greater proportion of cows in this group had hypocalcemia when compared with cows without clinical metritis. The circulating concentration of leukocytes and erythrocytes were decreased in cows with clinical metritis compared with cows without clinical metritis. Results from this study showed that concentrations of markers of inflammation, stress, pain, and activity were affected in cows diagnosed with clinical metritis; thus, strategies aimed to minimize the negative effects associated with clinical metritis may be required to improve the welfare of dairy cows.

The objectives of this study were to assess the responses to treatments (clinical cure and cow survival 14 d posttherapy) of cows with clinical endometritis (CE) that received intrauterine infusion of a hypertonic solution of 50% dextrose (DEX) or subcutaneous ceftiofur crystalline free acid (CCFA) and subsequent pregnancy per artificial insemination (P/AI) in cows with CE compared with cows without CE. Cows (n=760) from 2 dairy herds were screened for CE using vaginoscopy and measurement of cervix diameters [exam 1; 26±3 d in milk (DIM)]. Cows with vaginal discharge scores of 2 or 3 (scale 0-3) were stratified by parity and randomly allocated into 1 of 3 treatment groups: (1) intrauterine infusion (∼200 mL) of 50% DEX solution (n=79); (2) 6.6 mg/kg single-dose of subcutaneous administration of CCFA (n=75); or (3) untreated control animals (CON, n=83). Fourteen days posttherapy (at 40±3 DIM), cows with CE were re-examined (exam 2; 40±3 DIM) to assess the response to treatments. All cows were presynchronized with 2 injections of PGF(2α) given 14 d apart (starting at 26±3 DIM) followed by Ovsynch (OV; GnRH-7 d-PGF-56 h-GnRH 16 h-timed-AI) 12 to 14 d later. Cows displaying signs of standing estrus any time during the protocol were inseminated, whereas the remaining cows were subjected to timed AI 16 h after the second GnRH of OV. Pregnancy diagnosis was performed via transrectal ultrasonography at 39±3d post-AI followed by pregnancy reconfirmation 30 d after the first pregnancy diagnosis. Uterine swabs revealed that Arcanobacterium pyogenes and Escherichia coli were the most predominant bacteria isolated at the time of treatments. Mortality within 14 d posttherapy was not different among treatment groups. Cows with CE had greater cervical diameter at exam 1 and decreased P/AI compared with cows without CE. Treatment with CCFA or DEX increased the proportion of cows with clear vaginal discharge (score 0; clinical cure) 14 d posttherapy compared with CON cows. Pregnancy per AI from DEX (29.8±4%) cows tended to differ from that of CON (21.1±4%) or CCFA cows (19.7±4%), but it resulted in similar P/AI as those cows without CE (39.1±2%). The use of intrauterine DEX alone or as an adjunct of antibiotic therapy for the treatment of CE needs further investigation.