Sire contribution to fertilization failure and early embryo survival in cattle

Despite passing routine laboratory tests of semen quality, bulls used in artificial insemination (AI) exhibit a significant range in field fertility. The objective of this study was to determine whether subfertility in AI bulls is due to issues of sperm transport to the site of fertilization, fertilization failure, or failure of early embryo or conceptus development. In experiment 1, Holstein-Friesian bulls (3 high fertility, HF, and 3 low fertility, LF) were selected from the national population of AI bulls based on adjusted fertility scores from a minimum of 500 inseminations (HF: +4.37% and LF: ?12.7%; mean = 0%). Superovulated beef heifers were blocked based on estimated number of follicles at the time of AI and inseminated with semen from HF or LF bulls (n = 3–4 heifers per bull; total 19 heifers). Following slaughter 7 d later, the number of corpora lutea was counted and the uteri were flushed. Recovered structures (oocytes/embryos) were classified according to developmental stage and stained with 4′,6-diamidino-2-phenylindole to assess number of cells and accessory sperm. Overall recovery rate (total structures recovered/total corpora lutea) was 52.6% and was not different between groups. Mean (± standard error of the mean) number of embryos recovered per recipient was 8.7 ± 5.2 and 9.4 ± 5.5 for HF and LF, respectively. Overall fertilization rate of recovered structures was not different between groups. However, more embryos were at advanced stages of development (all blastocyst stages combined), reflected in a greater mean embryo cell number on d 7 for HF versus LF bulls. Number of accessory sperm was greater for embryos derived from HF than for LF bulls. The aim of experiment 2 was to evaluate the effect of sire fertility on survival of bovine embryos to d 15. Day 7 blastocysts were produced in vitro using semen from the same HF (n = 3) and LF (n = 3) bulls and transferred in groups of 5–10 to synchronized heifers (n = 7 heifers per bull; total 42 heifers). Conceptus recovery rate on d 15 was higher in HF (59.4%,) versus LF (45.0%). Mean length of recovered conceptuses for HF bulls was not affected by fertility status. In conclusion, while differences in field fertility among AI sires used in this study were not reflected in fertilization rate, differences in embryo quality were apparent as early as d 7. These differences likely contributed to the higher proportion of conceptuses surviving to d 15 in HF bulls.     

Use of intravaginal progesterone-releasing inserts in a synchronization protocol before timed AI and for synchronizing return to estrus in Holstein heifers

Holstein heifers (n = 189) were submitted to a 42-d artificial insemination (AI) period in which they underwent AI after once-daily evaluation of rubbed tail chalk. At the onset of the AI period (d 0), heifers were assigned randomly to receive synchronization of ovulation and timed AI (TAI; d 0: 100 mug of GnRH; d 6: 25 mg of PGF(2alpha); d 8: 100 mug of GnRH + TAI) either without (GPG; n = 95), or with inclusion of a CIDR insert (CIDR; n = 94) from d 0 to 6. No CIDR heifers received AI before d 8 compared with 24% of GPG heifers, and pregnancy rate per AI (PR/AI) at 30 d after TAI did not differ between treatments. To synchronize return to estrus for heifers failing to conceive after TAI, heifers (n = 166) receiving TAI to first service were randomly assigned to receive no further treatment (control; n = 85) or receive a new CIDR insert between 14 and 20 d after TAI (Resynch; n = 81). No Resynch heifers received AI during CIDR treatment compared with 35% of control heifers, and the proportion of heifers receiving AI within 72 h after the day of CIDR removal was 78 vs. 50% for Resynch vs. control heifers, respectively. No treatment x inseminator interaction was detected at first or second AI; however, overall PR/AI was modest for heifers throughout the experiment due to poor performance of 2 of the 3 herd inseminators (14, 6, and 58% PR/AI, respectively). Inclusion of CIDR inserts suppressed estrus during the TAI protocol with no reduction in PR/ AI. Resynchronization of estrus using CIDR inserts resulted in tighter synchrony of estrus among nonpregnant heifers compared with untreated controls.     

Evaluation of reproductive performance in lactating dairy cows with prostaglandin F2alpha, gonadotropin-releasing hormone, and timed artificial insemination

Nine hundred and twenty Holstein cows from 16 commercial dairy herds to evaluate three systematic breeding protocols: 14-d PGF2alpha, timed artificial insemination (AI), and GnRH-PGF2alpha, relative to AI following estrus detection without hormone intervention. The timed AI protocol involved GnRH, followed by PGF2alpha 7 d later and GnRH again 2 d after PGF2alpha, with AI 6 to 18 h after the second GnRH. The GnRH-PGF2alpha protocol consisted of GnRH followed by PGF2alpha 7 d later. Eight herds relied on visual observation to detect estrus, and eight herds utilized the HeatWatch estrus detection system. The average interval to first postpartum AI was shortest for the timed AI protocol (77.1 d) followed by the 14-d PGF2alpha protocol (81.6 d). There was no difference in days to first AI between the control (86.1 d) and GnRH-PGF2alpha (89.5 d) protocols. Percent pregnant per first AI did not differ among control (45.6%), 14-d PGF2alpha (43.7%), or GnRH-PGF2alpha (44.0%) protocols, but all protocols had a higher percent pregnant per first AI than the timed AI protocol (30.1%). Response to the GnRH-PGF2alpha protocol was limited because 44.0% of the cows submitted to the protocol were not detected in estrus < or = 10 d post-PGF2, administration and had an interval to first AI of 103.8 d. Cumulative percent pregnant by 120 d postpartum did not differ between control cows (53.1%) and hormonally treated cows (50.6%). Visual observation herds had a shorter interval to first postpartum AI (82.8 d) than the HeatWatch herds (84.8 d), with a higher overall rate of estrus detection across all protocols (75.3 and 67.6%, respectively).     

Inseminations at estrus induced by presynchronization before application of synchronized estrus and ovulation

A controlled field study examined conception rates after 2 timed artificial insemination (TAI) breeding protocols conducted on 2 commercial dairy farms. Estrous cycles in postpartum lactating cows were presynchronized with 2 injections of PGF(2alpha) given 14 d apart (Pre-synch) and then, after 12 d, the standard Ovsynch protocol (injection of GnRH 7 d before and 48 h after an injection of PGF(2alpha), with one TAI at 12 to 16 h after the second GnRH injection) or Heatsynch protocol [injection of GnRH 7 d before an injection of PGF(2alpha), followed 24 h later by 1 mg of estradiol cypionate (ECP) and one TAI 48 h after ECP] was applied. Experimental design allowed artificial insemination to occur anytime after the second Presynch injection and during the designed breeding week when estrus was detected. Of the 1846 first services performed, only 1503 (rate of compliance = 81.4%) were performed according to protocol. Numbers of cows inseminated, logistic-regression adjusted conception rates, and days in milk (DIM) were for inseminations made: 1) during 14 d after first Presynch injection (n = 145; 22.6%; 54 +/- 0.4 DIM); 2) during 12 d after second Presynch injection (n = 727; 33%; 59 +/- 0.2 DIM); 3) during 7 d after the first GnRH injection of Ovsynch or Heatsynch (n = 96; 32.1%; 74 +/- 0.5 DIM); 4) after estrus as part of Heatsynch (n = 212; 44.6%; 76 +/- 0.3 DIM); 4) after TAI as part of Heatsynch (n = 154; 21.1%; 76 +/- 0.4 DIM); 5) after estrus as part of Ovsynch (n = 43; 48.7%; 77 +/- 0.7 DIM); and 6) after TAI as part of Ovsynch (n = 271; 24.4%; 77 +/- 0.3 DIM). Conception rates when AI occurred after one Presynch injection were less than when AI occurred after 2 Presynch injections. Conception rates for those inseminated after either Presynch injection did not differ from those inseminated after combined Heatsynch + Ovsynch. Cows in the Ovsynch and Heatsynch protocols inseminated after estrus during the breeding week had greater conception rates than those receiving the TAI, but overall conception rates did not differ between protocols. Among cows inseminated after detected estrus, conception was greater for cows in the Heatsynch + Ovsynch protocol (77 +/- 0.4 DIM) than for those inseminated after either Presynch injection (54 +/- 0.4 or 59 +/- 0.2 DIM). We concluded that conception rates after Heatsynch and Ovsynch were similar under these experimental conditions, and that delaying first AI improved fertility for cows inseminated after detected estrus.     

Estrous detection intensity and accuracy and optimal timing of insemination with automated activity monitors for dairy cows

The objectives of this observational study were to assess the ability of automated activity monitoring (AAM) to detect estrus for first insemination, the accuracy of detection, and the optimum interval from the estrus alert from the AAM system to insemination. Four commercial farms using 1 of 2 commercial AAM systems were studied over 1 yr. Cows were inseminated between 55 and 80 d in milk (DIM) based on AAM only, then by a combination of AAM and timed artificial insemination (AI). Blood progesterone was measured in 1,014 cows at wk 5, 7, and 9 postpartum; purulent vaginal discharge (PVD) was assessed at wk 5; and lameness and BCS at wk 7. Overall, AAM detected 83% of cows in estrus by 80 DIM. Cows that had 3 serum progesterone <1 ng/mL, had PVD, or were both lame and had BCS ≤2.5 has lesser odds of being detected in estrus by 80 DIM (62, 68, and 53%, respectively). Blood samples were collected on the day of 445 AI based on AAM and 323 timed AI. The proportion of cows not in estrus (progesterone >1 ng/mL) on the day of AI was similar between AAM (4 ± 1.8%) and timed AI (3 ± 1.2%). Managers elected, based on subjective criteria, not to inseminate 17% of cows for which an AAM estrus alert was issued, of which 43% were not in estrus. Activity data were extracted from AAM software for 1,399 AI. Onset of estrus was calculated using the same or similar data processing criteria as the AAM system. Producers recorded the time of AI. The interval from onset of estrus to AI was categorized as 0 to 8, 8 to 16, or 16 to 24 h. We found no effect of AAM system on the probability of pregnancy per AI, but noted an interaction of interval with parity. For multiparous cows, the probability of pregnancy per AI was 31%, which did not differ with the interval to AI. For primiparous cows, the odds of pregnancy were greater if AI occurred 0 to 8 h (49%) than 8 to 16 (36%) or 16 to 24 h (31%) after the estrus alert from the AAM. Automated activity monitoring can detect estrus for first AI in just over the length of 1 estrous cycle for over 80% of cows, but the remainder would likely require intervention for timely insemination. For multiparous cows, performing AI based on AAM once per day would not affect pregnancy per AI, but for primiparous cows AI within 8 h of the onset of estrus may be advantageous.     

Resynchronizing estrus and ovulation after not-pregnant diagnosis and various ovarian states including cysts

We compared outcomes of 2 protocols used to resynchronize estrus and ovulation in dairy females after a not-pregnant diagnosis. Nulliparous heifers and lactating cows in which artificial insemination (AI) occurred 41 +/- 1 d earlier were presented every 2 to 3 wk for pregnancy diagnosis by using ultrasonography. Ovaries were scanned, follicles were mapped and sized, presence of corpus luteum was noted, and GnRH was injected (d 0). Females were assigned randomly to receive PGF(2alpha) 7 d later (d 7) and then either received estradiol cypionate (ECP) 24 h after PGF(2alpha) (d 8; Heatsynch; n = 230) or a second GnRH injection 48 h after PGF(2alpha) (d 9; Ovsynch; n = 224). Those detected in estrus since their not-pregnant diagnosis were inseminated, whereas the remainder received a timed AI (TAI) between 65 and 74 h after PGF(2alpha). Ovarian scans and blood collected before injections for progesterone analysis were used to classify 4 ovarian status groups: anestrus, follicular cysts, luteal cysts, and cycling, plus an unknown group of females in which no blood sampling or ovarian scans were made. Few females (5.1%) were inseminated between not-pregnant diagnosis and d 8. On d 10, more ECP- than GnRH-treated females were inseminated after detected estrus (24 vs. 6%). Overall, more Ovsynch than Heatsynch females received a TAI (82 vs. 62%). Conception rates tended to be greater for females inseminated after estrus (37%) than after TAI (29%), particularly for those treated with Heatsynch (41 vs. 27%) than with Ovsynch (33 vs. 31%). Those inseminated after estrus conceived 31 +/- 8 d sooner than those receiving the TAI. Conception rates for females having elevated progesterone 7 d after the not-pregnant diagnosis were greater than those having low progesterone in Heatsynch (42%; n = 133 vs. 25%; n = 55) and Ovsynch protocols (33%; n = 142 vs. 15%; n = 45), respectively. Conception rates were greater in nulliparous heifers than in lactating cows (43 vs. 28%) regardless of protocol used. Although overall pregnancy outcomes after a not-pregnant diagnosis were similar in response to either the Ovsynch and Heatsynch protocols, inseminations performed after detected estrus before the scheduled TAI reduced days to eventual conception and tended to increase conception rates, particularly after Heatsynch.     

Controlling first service and calving interval by prostaglandin F2 alpha, gonadotropin-releasing hormone, and timed insemination

Our objective was to determine if calving intervals could be shortened and made less variable by using prostaglandin F2 alpha to control the occurrence of first services. Holstein cows (n = 348) were assigned at calving to four treatment groups. Control cows (n = 88) were inseminated at their first observed estrus after 40 d postpartum. Estrous cycles of the remaining cows were synchronized with prostaglandin F2 alpha to allow insemination (first services) 80 h after the second injection (n = 86), insemination at 80 h preceded by gonadotropin-releasing hormone at 72 h (n = 86), or insemination at 72 and 96 h (n = 88) after the second injection (51 to 57 d postpartum). By design, interval to first service was reduced to 57 d for treated cows (63 d for controls) and was less variable (12% of that for controls). Conception rate at first service was lower after timed inseminations than that of controls. Intervals to conception and subsequent calving were similar in all treated cows and controls. Of cows sampled, 23 of 176 (13%) failed to respond with luteolysis when progesterone in serum exceeded 1 ng/ml, and 26 of 176 (15%) had low concentrations of progesterone in serum and could not respond to prostaglandin F2 alpha. Poor response to timed inseminations may have occurred because only 72% of 176 cows sampled responded with luteolysis. When only cows observed in estrus were considered, conception rate approached that of controls (51%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Gonadotropin-releasing hormone and conception of Holsteins

Effects of gonadotropin-releasing hormone (100 micrograms) and time of artificial insemination on fertility, were examined in lactating dairy cows at first, second, and third services. Inseminations were either soon after detected estrus (0 h) or 12 h later, and cows were given gonadotropin-releasing hormone or saline after inseminations. Conception at first service was not improved by hormone treatment. But conception rates at repeat services were improved by nearly 21% when cows were treated with gonadotropin-releasing hormone within 30 s after insemination. Inseminations soon after detection of estrus (0 h) followed by hormone treatment resulted in the highest conception rates for all services. Time of the insemination (0 versus 12 h) had no effect on conception. Administering gonadotropin-releasing hormone at repeat services should improve conception rate of lactating dairy cows.     

Role of myometrial activity in sperm transport through the genital tract and in fertilization in sows

The effects of stimulation and suppression of uterine contractility at about the time of insemination on sperm distribution and fertilization in multiparous sows are described. For assessment of fertilization, sows were inseminated about 28 h before (synchronized) ovulation and killed at day 5 after ovulation (n = 53). For assessment of sperm distribution, sows were inseminated about 20 h before expected ovulation and were killed 12 h later (n = 26). At 10 min before insemination, sows received an intrauterine infusion of one of three solutions: (i) saline (control); (ii) 0.60 mg clenbuterol hydrochloride to suppress contractility; or (iii) 1 mg cloprostenol to stimulate contractility. Both clenbuterol and cloprostenol reduced median fertilization rate (P < 0.05) and median number of accessory sperm cells (P < 0.05). Distribution of sperm cells was also affected by treatments. Clenbuterol increased, and cloprostenol decreased, the number of sperm cells (P < 0.05) in the proximal 20 cm of the uterine horn and in the uterotubal junction. In addition, clenbuterol tended to increase and cloprostenol tended to decrease the number of sperm cells in the isthmus, although these effects were not significant. However, relative to the number of sperm cells in the uterus, clenbuterol treatment reduced the number of sperm cells in the uterotubal junction and oviduct, in contrast to cloprostenol. Cloprostenol increased the reflux of semen during insemination. It is hypothesized that suppression of uterine contractility increases transuterine transport time, reducing the ability of sperm cells to enter the uterotubal junction and the oviduct. Stimulation of uterine contractility above a certain level probably increases reflux and impedes transuterine transport of sufficient numbers of sperm cells.     

Short communication: Conception rates following detection of estrus and timed AI in dairy cows synchronized using GnRH and PGF2alpha

The objective of this study was to compare conception rates of cows exhibiting spontaneous estrus and receiving artificial insemination (AI) before completion of a timed AI protocol with cows that did not display estrus spontaneously, but were inseminated after 1 of 3 GnRH-PGF_2α protocols. Cows (n = 432) in 2 herds were administered GnRH on d -7 and were tail-chalked daily. Cows detected in estrus before d 0 were inseminated immediately. Cows not detected in estrus by d 0 were administered PGF_2α and were tail-chalked daily until 48 h after PGF_2α. Cows detected in estrus from d −7 to 48 h after PGF_2α were inseminated and designated as treatment A (n = 46). Cows not detected in estrus and not inseminated by 48 h after PGF_2α were assigned randomly to receive either GnRH 48 h after PGF_2α and timed AI 16 h later (treatment B; n = 132), or GnRH and timed AI 64 h after PGF_2α (treatment C; n = 127), or timed AI 64 h after PGF_2α (treatment D; n = 127). Pregnancy was diagnosed 38 to 45 d after AI by palpation per rectum of uterine contents. Nearly 11% of all cattle exhibited spontaneous estrus and received immediate AI. Herd did not influence the percentage of cows detected in estrus and inseminated. Conception rates did not differ among treatments. Conception rates differed between herds, but no interaction of herd × treatment was detected. No differences were detected between herds for days in milk, milk production, AI service number, or parity.     

Factors affecting conception rates following artificial insemination or embryo transfer in lactating Holstein cows

The objective of this study was to evaluate the factors that may affect conception rates (CR) following artificial insemination (AI) or embryo transfer (ET) in lactating Holstein cows. Estrous cycling cows producing 33.1 ± 7.2 kg of milk/d received PGF_2α injections and were assigned randomly to 1 of 2 groups (AI or ET). Cows detected in estrus (n = 387) between 48 and 96 h after the PGF_2α injection received AI (n = 227) 12 h after detection of estrus or ET (n = 160) 6 to 8 d later (1 fresh embryo, grade 1 or 2, produced from nonlactating cows). Pregnancy was diagnosed at 28 and 42 d after estrus, and embryonic loss occurred when a cow was pregnant on d 28 but not pregnant on d 42. Ovulation, conception, and embryonic loss were analyzed by a logistic model to evaluate the effects of covariates [days in milk (DIM), milk yield, body temperature (BT) at d 7 and 14 post-AI, and serum concentration of progesterone (P4) at d 7 and 14 post-AI] on the probability of success. The first analysis included all cows that were detected in estrus. The CR of AI and ET were different on d 28 (AI, 32.6% vs. ET, 49.4%) and 42 (AI, 29.1% vs. ET, 38.8%) and were negatively influenced by high BT (d 7) and DIM. The second analysis included only cows with a corpus luteum on d 7. Ovulation rate was 84.8% and was only negatively affected by DIM. Conception rates of AI and ET were different on d 28 (AI, 37.9% vs. ET, 59.4%) and 42 (AI, 33.8% vs. ET, 46.6%) and were negatively influenced by high BT (d 7). The third analysis included only ovulating cows that were 7 d postestrus. Conception rates of AI and ET were different on d 28 (AI, 37.5% vs. ET, 63.2%) and 42 (AI, 31.7% vs. ET, 51.7%) and were negatively influenced by high BT (d 7). There was a positive effect of serum concentration of P4 and a negative effect of milk production on the probability of conception for the AI group but not for the ET group. The fourth analysis was embryonic loss (AI, 10.8% vs. ET, 21.5%). The transfer of fresh embryos is an important tool to increase the probability of conception of lactating Holstein cows because it can bypass the negative effects of milk production and low P4 on the early embryo. The superiority of ET vs. AI is more evident in high-producing cows. High BT measured on d 7 had a negative effect on CR and embryonic retention.     

Impact of site of inseminate deposition and environmental factors that influence reproduction of dairy cattle

Uterine body and cornual inseminations (n = 2127) were evaluated over a 3-yr period in Holstein and Jersey cattle. For cornual insemination one-half of each semen dose was deposited approximately 2.5 cm into each uterine horn. Conception rate was lower for cervical insemination (39.4%) than uterine body (48.1%) or cornual (49.3%) inseminations. Pregnancy site distribution was equal for both insemination techniques but cervical insemination resulted in 60% right horn pregnancies. Primary housing, service number, inseminator, lactation number, and sire affected conception rate. Older cows were least fertile (31.4%), second service conception rate was lowest (42.6%), and barn housed cattle had a 39.7% conception rate. Days open was affected by primary housing, service number, sire, site of semen placement, and twinning. Twinning increased days open by 10 d. Optimum time for insemination of lactating cows was between 6 and 12 h after the initial observation of estrus. From this study we conclude that shallow cornual insemination is as effective as uterine body insemination, and conception rate is optimized when estrus is positively assessed.     

Influence of gonadotropin-releasing hormone and timing of insemination relative to estrus on pregnancy rates of dairy cattle at first service

The objective was to determine the influence of gonadotropin-releasing hormone on pregnancy rates of dairy cattle at first services, when both the timing of hormone injection and insemination were altered relative to the onset of estrus. Cows (n = 325) were assigned randomly to six groups making up a 2 X 2 X 2 incomplete factorial experiment; dose of GnRH (100 micrograms versus saline), timing [1 h (early) or 12 to 16 h (late) after first detected estrus] of AI, and timing of hormone injection (early versus late) were the three main effects. Cows were observed for estrus 4 times daily. Treatments and resulting pregnancy rates were: 1) hormone injection early plus AI early (35%), 2) hormone injection late plus AI early (34%), 3) saline injection early plus AI early (30%), 4) hormone injection late plus AI late (30%), 5) hormone injection early plus AI late (46%), and 6) saline injection late plus AI late (43%). Pregnancy rate in the first four groups (32%) was less than that in the latter two groups (44%). Concentrations of LH in serum were greater for cows given hormone or saline injections in early estrus than for cows injected with either hormone of saline during late estrus. Concentrations of LH in serum 2 h after GnRH were elevated above those of controls, whether GnRH was injected during early or late estrus. Neither concentrations of LH during estrus nor concentrations of progesterone 8 to 14 d after estrus explained the possible antifertility effect of GnRH given during late estrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of sire fertility and timing of artificial insemination in a Presynch + Ovsynch protocol on first-service pregnancy rates

Sire fertility may influence pregnancy rate (PR) by differences in sperm survival in the female reproductive system and time required for capacitation and transport of sperm to site of fertilization. A predicted fertility index, Estimated Relative Conception Rate, was used to select 3 high-fertility artificial insemination (AI) sires (+3) and 3 average AI sires (−1). Ovulation can be predicted to occur at approximately 28 h following GnRH administration when used in an Ovsynch protocol. The objective of this study was to determine if AI at 2 times, 0 or 24 h after GnRH administration, in a Presynch + Ovsynch protocol resulted in different first-service PR when average or high-fertility sires were used. Lactating Holstein cows (n = 1,457) from 2 dairy herds located in the Piedmont region of North Carolina were utilized for 12 mo. Timing of AI did not affect first AI PR and no interaction of sire-fertility group and timing of AI was detected. First AI PR did not differ between sire-fertility groups (23.2 vs. 29.4%) for average and high-fertility groups, respectively. First-lactation cows were 53% more likely to conceive than older cows, and cows bred during April through June were 66% less likely to become pregnant compared with cows bred from October through January. No interactions were detected among parity, season, sire-fertility group, or time of AI. Using only 3 sires per group based on Estimated Relative Conception Rate estimates resulted in large variability of sire conception within groups, although group averages differed by 6 points.     

Effects of gonadotropin-releasing hormone at initiation of the 5-d timed artificial insemination (AI) program and timing of induction of ovulation relative to AI on ovarian dynamics and fertility of dairy heifers

Two experiments evaluated the effects of the first GnRH injection of the 5-d timed artificial insemination (AI) program on ovarian responses and pregnancy per AI (P/AI), and the effect of timing of the final GnRH to induce ovulation relative to AI on P/AI. In experiment 1, 605 Holstein heifers were synchronized for their second insemination and assigned randomly to receive GnRH on study d 0 (n = 298) or to remain as untreated controls (n = 307). Ovaries were scanned on study d 0 and 5. All heifers received a controlled internal drug-release (CIDR) insert containing progesterone on d 0, a single injection of PGF_2α and removal of the CIDR on d 5, and GnRH concurrent with timed AI on d 8. Blood was analyzed for progesterone at AI. Pregnancy was diagnosed on d 32 and 60 after AI. Ovulation on study d 0 was greater for GnRH than control (35.4 vs. 10.6%). Presence of a new corpus luteum (CL) at PGF_2α injection was greater for GnRH than for control (43.1 vs. 20.8%), although the proportion of heifers with a CL at PGF_2α did not differ between treatments and averaged 87.1%. Progesterone on the day of AI was greater for GnRH than control (0.50 ± 0.07 vs. 0.28 ± 0.07 ng/mL). The proportion of heifers at AI with progesterone <0.5 ng/mL was less for GnRH than for control (73.8 vs. 88.2%). The proportion of heifers in estrus at AI did not differ between treatments and averaged 66.8%. Pregnancy per AI was not affected by treatment at d 32 or 60 (GnRH = 52.5 and 49.8% vs. control = 54.1 and 50.0%), and pregnancy loss averaged 6.0%. Responses to GnRH were not influenced by ovarian status on study d 0. In experiment 2, 1,295 heifers were synchronized for their first insemination and assigned randomly to receive a CIDR on d 0, PGF_2α and removal of the CIDR on d 5, and either GnRH 56 h after PGF_2α and AI 16 h later (OVS56, n = 644) or GnRH concurrent with AI 72 h after PGF_2α (COS72; n = 651). Estrus at AI was greater for COS72 than for OVS56 (61.4 vs. 47.5). Treatment did not affect P/AI on d 32 in heifers displaying signs of estrus at AI, but COS72 improved P/AI compared with OVS56 (55.0 vs. 47.6%) in those not in estrus at AI. Similarly, P/AI on d 60 did not differ between treatments for heifers displaying estrus, but CO S72 improved P/AI compared with OVS56 (53.0 vs. 44.7%) in those not in estrus at AI. Administration of GnRH on the first day of the 5-d timed AI program resulted in low ovulation rate and no improvement in P/AI when heifers received a single PGF_2α injection 5 d later. Moreover, extending the proestrus by delaying the final GnRH from 56 to 72 h concurrent with AI benefited fertility of dairy heifers that did not display signs of estrus at insemination following the 5-d timed AI protocol.     

Relationship among seminal quality measures and field fertility of young dairy bulls using low-dose inseminations

Optimal use of genetically superior bulls through artificial insemination (AI) is highly dependent on precise assessment of seminal quality which allows for reasonable estimations of field fertility with normal or low-dose inseminations. In the present study, seminal measures such as sperm motility and morphology, sperm viability, sperm DNA fragmentation, and the ability of the sperm to display an acrosome reaction were tested. The relationships between field fertility and the seminal measures were investigated using 3 ejaculates from each of 195 bulls (156 Holstein and 39 Jersey) participating in a progeny test program. A range of AI doses, varying from 2 × 10⁶ to 15 × 10⁶ sperm/straw, was obtained by a controlled dilution process applied to each ejaculate. The different AI doses were distributed at random among 75,610 experimental first inseminations in 4,721 herds and 208 AI technicians. Most of the seminal measures appeared to contain a predictive value for the nonreturn to estrus at 56 d post-AI (NRR56) regardless of the number of sperm per AI dose and can be regarded as noncompensable sperm traits. But, due to correlations between the individual measures, the best model for describing (and predicting) NRR56 was based on sperm concentration and viability in the neat (raw) semen, and post-thaw sperm viability. The statistical models for describing NRR56 included the following explanatory variables: strength of the estrus, number of sperm per AI dose, breed, parity, and random components representing herds and AI technicians. The present results show that the most precise estimation of a bull’s NRR56 can be achieved through flow cytometric detection of sperm concentration and viability in neat semen as well as flow cytometric detection of post-thaw sperm viability.     

Comparison of reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen based on expression of estrus,pregnancy outcomes,and cost per pregnancy

Our objective was to evaluate reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen. In experiment 1, nulliparous Holstein heifers (n = 462) were submitted to a 5-d progesterone-releasing intravaginal device (PRID)-Synch protocol [d 0, GnRH + PRID; d 5, PGF_2α − PRID; d 6, PGF_2α; d 8, GnRH + TAI] and were randomly assigned for PRID removal on d 5 or 6 of the protocol followed by timed artificial insemination (TAI) with conventional semen. Delaying PRID removal decreased early expression of estrus before scheduled TAI (0.9 vs. 12.2%), and pregnancies per AI (P/AI) did not differ between treatments. In experiment 2, nulliparous Holstein heifers (n = 736) from 3 commercial farms were randomized within farm to 1 of 3 treatments for first AI with sexed semen: (1) CIDR5 [d −6, GnRH + controlled internal drug release (CIDR); d −1, PGF_2α − CIDR; d 0, PGF_2α; d 2, GnRH + TAI]; (2) CIDR6 (d −6, GnRH + CIDR; d −1, PGF_2α; d 0, PGF_2α − CIDR; d 2, GnRH + TAI); and (3) EDAI (PGF_2α on d 0 followed by once-daily estrous detection and AI). Delaying CIDR removal decreased early expression of estrus before scheduled TAI (0.004 vs. 27.8%); however, CIDR5 heifers tended to have more P/AI at 35 (53 vs. 45 vs. 46%) and 64 (52 vs. 45 vs. 45%) days after AI than CIDR6 and EDAI heifers, respectively. Overall, CIDR5 and CIDR6 heifers had fewer days to first AI and pregnancy than EDAI heifers which resulted in less feed costs than EDAI heifers due to fewer days on feed until pregnancy. Despite greater hormonal treatment costs for CIDR5 heifers, costs per pregnancy were $16.66 less for CIDR5 than for EDAI heifers. In conclusion, delaying PRID removal by 24 h within a 5-d PRID-Synch protocol in experiment 1 suppressed early expression of estrus before TAI, and P/AI for heifers inseminated with conventional semen did not differ between treatments. By contrast, although delaying CIDR removal by 24 h within a 5-CIDR-Synch protocol in experiment 2 suppressed early expression of estrus before TAI, delaying CIDR removal by 24 h tended to decrease P/AI for heifers inseminated with sexed semen. Further, submission of heifers to a 5-d CIDR-Synch protocol for first AI tended to increase P/AI and decrease the cost per pregnancy compared with EDAI heifers.     

Fertility of holstein dairy heifers after synchronization of ovulation and timed AI or AI after removed tail chalk

Nonlactating Holstein dairy heifers (n=352) 13 mo of age were managed using a 42-d artificial insemination (AI) breeding period in which they received AI after removed tail chalk evaluated once daily. At AI breeding period onset (d 0), heifers were randomly assigned to receive synchronization of ovulation (100 microg of GnRH, d 0; 25 mg of PGF2alpha, d 6; 100 microg of GnRH, d 8) and timed AI (TAI; d 8) and AI after removed tail chalk for the entire AI breeding period (GPG; n=175), or AI after removed tail chalk for the entire AI breeding period (TC; n=177). As expected, 17.7% (31/175) of GPG heifers received AI after removed tail chalk before scheduled TAI. Pregnancy rate per artificial insemination (PR/AI) at approximately 30 d after first AI tended to be greater for TC (46.5%) than for GPG (38.3%) heifers. No treatment x inseminator interaction was detected; however, overall PR/AI was low for heifers in both treatments due to variation among the 3 inseminators (24.8, 30.0, and 58.0%). Pregnancy loss from approximately 30 to approximately 75 d after first AI was 10% and did not differ between treatments. Based on survival analysis, days to first AI was greater for TC than for GPG heifers, whereas days to pregnancy across the 42-d AI breeding period did not differ between treatments. Overall, 81.2% of GPG heifers receiving TAI synchronized luteal regression and ovulated within 48 h after the second GnRH injection. We conclude that this synchronization protocol can yield acceptable fertility in dairy heifers if AI to estrus is conducted between treatment with GnRH and PGF2alpha and AI efficiency is optimized.     

Fertilization and early embryonic development in heifers and lactating cows in summer and lactating and dry cows in winter

Two experiments in two seasons evaluated fertilization rate and embryonic development in dairy cattle. Experiment 1 (summer) compared lactating Holstein cows (n = 27; 97.3 +/- 4.1 d postpartum [dppl; 40.0 +/- 1.5 kg milk/d) to nulliparous heifers (n = 28; 11 to 17 mo old). Experiment 2 (winter) compared lactating cows (n = 27; 46.4 +/- 1.6 dpp; 45.9 +/- 1.4 kg milk/d) to dry cows (n = 26). Inseminations based on estrus included combined semen from four high-fertility bulls. Embryos and oocytes recovered 5 d after ovulation were evaluated for fertilization, embryo quality (1 = excellent to 5 = degenerate), nuclei/embryo, and accessory sperm. In experiment 1, 21 embryos and 17 unfertilized oocytes (UFO) were recovered from lactating cows versus 32 embryos and no UFO from heifers (55% vs. 100% fertilization). Embryos from lactating cows had inferior quality scores (3.8 +/- 0.4 vs. 2.2 +/- 0.3), fewer nuclei/embryo (19.3 +/- 3.7 vs. 36.8 +/- 3.0) but more accessory sperm (37.3 +/- 5.8 vs. 22.4 +/- 5.5/embryo) than embryos from heifers. Sperm were attached to 80% of UFO (17.8 +/- 12.1 sperm/UFO). In experiment 2, lactating cows yielded 36 embryos and 5 UFO versus 34 embryos and 4 UFO from dry cows (87.8 vs. 89.5% fertilization). Embryo quality from lactating cows was inferior to dry cows (3.1 +/- 0.3 vs. 2.2 +/- 0.3), but embryos had similar numbers of nuclei (27.2 +/- 2.7 vs. 30.6 +/- 2.1) and accessory sperm (42.0 +/- 9.4 vs. 36.5 +/- 6.3). From 53% of the flushings from lactating cows and 28% from dry cows, only nonviable embryos were collected. Thus, embryos of lactating dairy cows were detectably inferior to embryos from nonlactating females as early as 5 d after ovulation, with a surprisingly high percentage of nonviable embryos. In addition, fertilization rate was reduced only in summer, apparently due to an effect of heat stress on the oocyte.     

Supplementation with estradiol-17beta before the last gonadotropin-releasing hormone injection of the Ovsynch protocol in lactating dairy cows

The aim of this study was to determine whether an increase in circulating estrogen concentrations would increase percentage pregnant per artificial insemination (PP/AI) in a timed AI protocol in high-producing lactating dairy cows. We analyzed only cows having a synchronized ovulation to the last GnRH of the Ovsynch protocol (867/1,084). The control group (n = 420) received Ovsynch (GnRH - 7 d - PGF_2α - 56 h - GnRH - 16 h - timed AI). The treatment group (n = 447) had the same timed AI protocol with the addition of 1 mg of estradiol-17β (E2) at 8 h before the second GnRH injection. Ovarian ultrasound and blood samples were taken just before E2 treatment of both groups. In a subset of cows (n = 563), pressure-activated estrus detection devices were used to assess expression of estrus at 48 to 72 h after PGF_2α treatment. Ovulation was confirmed by ultrasound 7 d after timed AI. Treatment with E2 increased expression of estrus but overall PP/AI did not differ between E2 and control cows. There was an interaction between treatment and expression of estrus such that PP/AI was greater in E2-treated cows that showed estrus than in E2-treated or control cows that did not show estrus and tended to be greater than control cows that showed estrus. There was evidence for a treatment by ovulatory follicle size interaction on PP/AI. Supplementation with E2 improved PP/AI in cows ovulating medium (15 to 19 mm) but not smaller or larger follicles. The E2 treatment also tended to improve PP/AI in primiparous cows with low (≤2.5) body condition score, and in cows at first postpartum service compared with Ovsynch alone. In conclusion, any improvements in PP/AI because of E2 treatment during a timed AI protocol appear to depend on expression of estrus, parity, body condition score, and size of ovulatory follicle.