Analysis of cow factors influencing conception rates after two timed breeding protocols

The objective of this study was to determine cow factors that influence conception rates after timed artificial insemination (TAI) on a commercial dairy farm. In Trial 1, 197 cows were synchronized by an administration of 25 mg of dinoprost between Days 48 and 54 post partum and again 14 days later. Cows were inseminated 66 and 90 hours after the second treatment. In Trial 2, 186 cows were treated with an Ovsynch protocol consisting of an administration of 0.02 mg of buserelin between Days 62 and 68 post partum, a treatment with 0.75 mg of tiaprost 7 days later and a second treatment with buserelin 48 h later. Cows were bred 16 to 20 hours after the last treatment. Cows with abnormal vaginal discharge at the time of insemination were excluded from AI. Lactation number, milk yield, fat and protein content of milk, signs of endometritis at an examination 14 to 20 days post partum and month of breeding were included as potential factors influencing conception on TAI. Conception rates after timed breeding were 32.0% and 30.6% in Trials 1 and 2, respectively. Logistic regression revealed that neither milk production parameters nor endometritis at post partum examination influenced conception rates in either of the two timed breeding protocols. Only parity showed an effect (P=0.012) in Trial 2. Primiparous cows were more likely to conceive after timed breeding than older cows (43.5 vs. 23.1%). An effect of parity, however, was not observed in Trial 1. It can be concluded that neither milk production nor endometritis at an examination 14 to 20 days post partum influence conception rates after TAI if cows with abnormal vulval discharge at the designated time of AI are excluded from breeding.     

Diagnosis of pregnancy by radioimmunoassay of a pregnancy-specific protein in the plasma of dairy cows

The accuracy and efficiency of progesterone (P4) and bovine pregnancy-specific protein B (bPSPB) radioimmunoassays (RIA) in detecting pregnant and nonpregnant dairy cows were compared at different stages of pregnancy. The study included 145 French Friesian heifers and cows from a single herd. A total of 175 artificial insemination (A.I.) and blood sampling procedures were performed. Animals were bled 24 d post AI for P4 RIA. They were bled at 24, 26, 30 to 35, and 70 +/- 9 after AI for bPSPB RIA. Females were declared nonpregnant when plasma P4 concentrations were lower than 1.5 ng/ml. With the bPSPB RIA, cows were nonpregnant when at least one of the B Bo x 100 replicates was higher than 95% in the RIA. When compared with palpations per rectum at 70 d, the accuracy of positive diagnoses (no. positive and pregnant/no. positive diagnoses) by P4 RIA at Day 24 was 67.2% (82 122 ). The accuracy of negative diagnoses was 98% (52 53 ). Accuracy of positive diagnoses by bPSPB RIA increased with gestation age (P<0.05) from 86.2% (50 58 ) on Day 24 to 98.8% (83 84 ) at time of palpation per rectum. Accuracy of negative diagnoses increased (P< 0.001) from Day 24 (71.8%; 84 117 ) to Days 30 to 35 (100%, 83 83 ). Efficiency in detecting nonpregnant females was much higher (P < 0.001) with the bPSPB RIA on Days 30 to 35 (90.2%; 83 92 ) than with the P4 RIA on Day 24 (56.5%, 52 92 ). It is concluded that 30 days after AI, the bPSPB RIA is an efficient test both for pregnancy prediction and detection of nonpregnant dairy cows.     

Increase in milk and body temperature of cows as a sign of embryo entry into the uterus

The objective of the first stage of these studies was to investigate whether temperature increases in the milk and body of cows during the early period of pregnancy. We studied 94 pregnant and 116 nonpregnant cows, and the temperatures were measured daily beginning 24 days after insemination. In addition, progesterone levels in milk were measured twice (on Days 21 and 24 after insemination), and examinations per rectum were conducted to determine pregnancy. Data analysis showed that in almost 90% of the pregnant cows milk temperature increased by 0.64 degrees C some time between Days 5 and 12 after insemination. This increase in milk temperature was highly significant and corresponded with a 0.46 degrees C increase in body temperature. There was no increase in the milk or body temperature in nonpregnant cows. After discovering in the first stage that a high percentage of pregnant cows experienced both milk and body temperature increases, we focused our attention in the second stage of the study on the question of whether this increase in temperatures might be due to an immune response of the mother to the entry of the embryo into the uterus. In this stage we conducted three different experiments on another group of animals consisting of 309 cows and heifers. The experiments included analysis of progesterone, estradiol, cortisol and PGE2 levels in the blood serum of the cows and heifers; an estimation of the pyrogenic activity of PGE1, PGE2 and progesterone; and measurements of body temperature in the heifers before and after embryo transfer. The results of these experiments suggested that an increase in milk and body temperature could be an indicator of an immune response of the mother to the entry of the embryo into the uterus.     

Early pregnancy diagnosis in cattle by means of linear-array real-time ultrasound scanning of the uterus and a qualitative and quantitative milk progesterone test

We compared three methods for diagnosing early pregnancy in cattle: 1) a trans-rectal ultrasound scan of the uterus, 2) a cow-side enzymeimmunoassay (EIA) milk progesterone test 3) a radioimmunoassay (RIA) milk progesterone test. Scanning of the uterus was performed in 148 cows. These cows were not detected in estrus before scanning, which took place between Days 21 and 33 after insemination (AI). A considerable difference was noted between the reliability of the scannings performed at an early stage (Days 21 to 25) and those performed at a later stage (Days 26 to 33). The sensitivity and specificity of the ultrasound examination between Days 21 and 25 were only 44.8% and 82.3%, respectively, but were 97.7% and 87.8% between Days 26 and 33, respectively. Milk samples were collected on the day of AI. (Day 0) and 21 days later. Samples that were positive in the EIA test always contained more than 1 ng/ml progesterone (P4); however, 20% of the negative EIA samples contained also more than 1 ng/ml P4. Only 59% of the animals showing a negative EIA test on Day 0 and a positive test on Day 21, indicating pregnancy, calved, while 16% of the cows with a negative test on Day 0 and Day 21, indicating nonpregnancy, turned out to be pregnant. Of the 82 animals with P4 levels lower than 1 ng/ml on Day 0 and higher than 1 ng/ml on Day 21, only 61.0% calved. All 14 cows with low levels both on Day 0 and Day 21, indicating nonpregnancy, were found to be not pregnant. The influence of both early embryonic death and the accumulation of intrauterine fluids on the accuracy of these tests are discussed.     

Estrus synchronization in cattle using estradiol, melengestrol acetate and PGF

In Experiment 1, all cattle were fed MGA (0.5 mg/head/d) for 7 d (designated Days 0 to 6) and given PGF on Day 6. One-half were administered estradiol valerate (EV; 5 mg, im) on Day 0. At Location 1, a higher proportion (P < 0.005) of EV-treated heifers were detected in estrus and bred by AI between Days 7 and 13 than control heifers not receiving EV (27 of 33 versus 15 of 32), but the number of pregnancies (12 vs 10) was not significantly different. Eighty-three of 104 EV-treated and 89 of 106 control cows were inseminated, resulting in 50 and 45 pregnancies, respectively (not significant). At Location 2, cattle were similarly treated and exposed to bulls on Days 7 to 49. Fall pregnancy rate was higher (P < 0.015) for EV-treated than control heifers (44 of 48 vs 33 of 46), but was not significantly different for cows (22 of 26 vs 19 of 23). In Experiment 2, estradiol 17beta (E17beta; 5 mg, im) and progesterone (100 mg, im) were administered on Day 0 (instead of EV). In a third group (designated the PGF group), cattle were bred on Days 0 to 6, and PGF was administered on Day 6 to those not yet bred. For 213 cows, the percentage pregnant to a synchronized estrus was greater in the PGF group (72%) than in either the control group treated with MGA (49%; P = 0.005) or the group receiving MGA and E17beta (54%; P < 0.025). Fall pregnancy rates were 91, 89, and 96% for the 213 cows (not significant) and 89, 93, and 98% for 131 heifers (not significant) in the PGF, MGA and E17beta groups, respectively. In cattle without a functional CL, the average diameter of the largest follicle at Day 6 was 1 to 2 mm smaller in the E17beta + MGA group than in the MGA group (difference significant only in cows at Location 1). Combined for both locations, the synchronized pregnancy rate in heifers without a functional CL on Day 6 was higher (P < 0.05) in the E17beta + MGA group than in the MGA group (11 of 21, 52% versus 4 of 20, 20%). Estrogen treatment caused regression of ovarian follicles with emergence of a new follicular wave. Including estrogen in an estrus synchronization program utilizing MGA and PGF significantly increased fall pregnancy rate in heifers (at 1 location) and the synchronized pregnancy rate of heifers without a functional CL at the time of PGF treatment (combined for both locations).     

Accuracy of ultrasonography for pregnancy diagnosis on days 10 to 22 in heifers

Two operators independently conducted ultrasonic pregnancy examinations on nulliparous Holstein heifers on Days 10, 12, 14, 16, 18, 20 and 22, and assigned a diagnosis (pregnant or nonpregnant) and a score for degree of certainty in the diagnosis (1, 2 or 3 for low, intermediate or high, respectively). Pregnancy was retrospectively confirmed by the ultrasonographic detection of an embryo proper and embryonic heartbeat on Day 24 in 20 25 bred heifers; the five nonpregnant heifers were excluded from the analyses. Eleven nonbred heifers were included as an unequivocal source of nonpregnant heifers. Accuracy was not significantly greater than a guess (50%) before Day 18, but reached 100% on Days 20 and 22. Mean accuracy was higher (P<0.005) for nonpregnant (65 77 , 84%) than pregnant heifers (91.5 140 , 65%). For certainty score, there were main effects of day (P<0.0001), reproductive status (pregnant or nonpregnant, P<0.003), and an interaction of day and reproductive status (P<0.0001). The certainty score increased in all heifers among days and was higher (P<0.05) in pregnant than nonpregnant heifers on Days 16 to 20. For luteal area (area of corpus luteum, excluding area of fluid filled center, if present), there were significant main effects of day, reproductive status and a day by status interaction (P<0.0001 for each). Luteal area was approximately constant in pregnant heifers, but in nonpregnant heifers it was lower (P<0.05) on Days 16 to 22 than on Days 10 to 14. Uterine echotexture was scored on a scale of 1 to 3, characteristic of a diestrus, intermediate and estrus uterus, respectively. There were main effects of day and reproductive status (P<0.0001 for each) and an interaction of day and reproductive status (P<0.025). Uterine echotexture was approximately constant in pregnant heifers, but in nonpregnant heifers it was higher (P<0.05) on Days 16 to 22 than on Days 10 to 14. Pregnancy diagnosis on Days 10 to 14 was based on detection of the conceptus; however, detection of the conceptus was not accurate prior to visualization of the embryo proper (mean Day 22, range Days 20 to 24). In nonpregnant heifers, a correct diagnosis with high certainty was made when a small corpus luteum and uterine echotexture characteristic of estrus were detected. In the absence of these changes on Days 18 to 22, a diagnosis of pregnancy was made with high accuracy and intermediate or high certainty.     

Artificial insemination of subtropical commercial beef cattle following synchronization with cloprostenol (ICI 80996): I. Fertility

Two trials were conducted over a two-year period with 519 cycling Bos taurus x Bos indicus heifers and cows. The objectives of these trials were: 1) To compare fertility of artificial insemination at the cloprostenol-induced estrus and the naturally occurring estrus, 2) To evaluate the fertility of artificial insemination at a predetermined time (Timed AI) following an estrous synchronization regime with cloprostenol (CLP) and 3) To define the optimum interval from a second CLP treatment for Timed AI. In Trial I, 128 animals were assigned to four treatments: 1) Controls, which were inseminated at the natural occurring estrus; 2) timed AI at 72 hr and again at 96 hr post-second CLP; 3) Timed AI at 72 hr post-second CLP and 4) AI at the CLP-induced estrus. Trial II included 391 heifers distributed among six treatments; 1) Timed AI between 70 and 90 hr post-second CLP; 2) Sham AI between 70 and 90 hr post-second CLP, 3) Chute Stress between 70 and 90 hr post-second CLP; 4) AI at the CLP-induced estrus; 5) Control-AI at the naturally occurring estrus and 6) Non-treated and exposed to fertile bulls. The fertility of the animals artificially inseminated at the CLP-induced estrus was similar to that of insemination at the naturally occurring estrus in Trial I and Trial II (30 vs 46%; 37 vs 38%, respectively). The first service pregnancy rates of the animals bred at a predetermined time were similar to those bred at the CLP-induced estrus in Trial I, but lower in Trial II (P < .01).     

Reproductive performance of lactating dairy cows following estrus synchronization regimens with PGF2alpha and progesterone

The objective of this study was to evaluate the reproductive performance of lactating cows in seasonal dairy herds after estrus synchronization with PGF2alpha (PG) with or without supplementation with progesterone (P4). In Trial 1, synchronized cows (S1; n = 521) were compared with untreated control cows (C; n = 518) in 5 herds. Estrus of cows in the S1 group was synchronized with 2 treatments of PG (Lutalyse) 13 d apart. The breeding season started 2 d after the second PG. Cows were first bred by AI for 7 wk and then herd sires were used. Compared with C cows, estrus synchronization in the treated cows reduced the conception rate to first AI (61.1 vs 70.5%; P < 0.01) and the intervals from start of the breeding season to conception for cows conceiving to AI (11.0 vs 14.6 d; P < 0.05) or to both AI and natural mating (16.5 vs 18.4 d; P < 0.05). There was no effect on conception rate to second AI (68.8%), on pregnancy rate by Day 24 (72.3%) or Day 49 (86.3%) of the breeding season, or on the percentage of cows not pregnant at end of the breeding season (5.0%). In Trial 2, effects of P4 supplementation before the second PG on reproductive performance were evaluated in 4 herds. Estrus of each cyclic cow was synchronized with PG as in Trial 1. Half of the cows in each herd were treated with an intravaginal P4 device (CIDR) for 5 d before the second PG (S2+P4, n = 608), whereas the remaining half received no CIDR treatment (S2, n = 593). Compared with S2 cows, P4 treatment increased the estrous response rate to the second PG (89.6 vs 82.9%; P < 0.01), the conception rate to first AI (65.1 vs 59.7%; P = 0.07), the pregnancy rate by Day 6 of the breeding season (59.3 vs 49.0%; P < 0.001), and reduced the intervals from start of the breeding season to conception for cows conceiving to AI (8.6 vs 10.4 d; P < 0.10) or to both AI and natural mating (12.7 vs 16.4 d; P < 0.01). Treatment with a used CIDR from Days 16 to 21 after start of breeding to re-synchronize returns to service had no effect on conception rate to first or second AI but may decrease the conception rate to second AI in cows previously treated with CIDR. In conclusion, estrus synchronization with the double PG system can reduce fertility, while P4 supplementation for 5 d before the second PG can improve estrous response and overall reproductive performance. Stage of the estrous cycle at the time of the second PG can affect fertility following synchronization.     

Plasma progesterone profiles and fertility status of anestrus Zebu cattle treated with norgestomet-estradiol-eCG regimen

Zebu cattle are notorious for poor fertility characterized by late maturity and long intercalving intervals attributed to a variety of factors, including genetic, nutritional and climatic. The aim of the present investigation, therefore, was to induce fertile estrus in acyclic pubertal heifers and postpartum anestrous Zebu cows by hormonal intervention. Pubertal Hariana and Sahiwal anestrous heifers (n=51) and postpartum cows (n=55) were either assigned a placebo (controls, N=6 for each breed and parity) or treated with 10-d norgestomet (3 mg) subcutaneous ear implants, with an initial injection of 3 mg, im norgestomet + 5 mg estradiol valerate, followed by 500 IU eCG at implant withdrawal (NOR-treated groups). Jugular venous plasma samples were obtained from a total of 28 animals (controls : 4 heifers and 4 cows; NOR-treated : 12 heifers and 8 cows) on Days 0 (implant insertion), 3, 7, 9 and Day 10 (implant withdrawal), every 12 h on Days 11 and 12, and then once daily on Days 17, 24 and 31. All the samples were assayed for progesterone. Almost all (97%) heifers and 81% cows were induced to estrus, the majority (92% heifers and 79% cows) within 120 h of implant removal. Synchrony of the induced estrus was better in cows, but interval to estrus and estrus duration were significantly longer in heifers (P<0.05). Post-treatment fertility, based on Day 28 nonretum rate, first service, and overall conception rates, was better in heifers (78.9, 60.5 and 73.7%, respectively) than cows (77.1, 48.6 and 62.9%, respectively), but the differences were significant only for the overall pregnancy rate (71.8% for heifers and 51.2% for cows; P<0.05). Low pre-treatment plasma progesterone values (<0.5ng/mL) were consistent with ovarian inactivity, confirming the true anestrus status of experimental animals. Controls failed to exhibit estrus and maintained low progesterone concentrations throughout the study. In treated animals, high progesterone values from Day 17 onwards suggested ovulatory estrus. These early luteal phase progesterone concentrations in nonpregnant (P=0.06) and nonpregnant, nonretum (P<0.05) animals were low in comparison with those of pregnant animals. Good fertility resulting from breeding according to estrus, inspite of variable intervals to estrus and estrus duration, advocates its advantage over fixed-time insemination in norgestomet-treated anestrous Zebu cattle.     

Conception, embryonic development and corpus luteum function in beef cattle open for two consecutive breeding seasons

High-fertility (control cows) and low-fertility (cows and heifers not pregnant after two consecutive breeding seasons — twice-open) cyclic bovine females were treated with a single injection of 1000 IU of human chrionic gonadotropin (HCG) or 100 μg of gonadotropin releasing hormone (GnRH) to enhance and/or hasten corpus luteum formation and progesterone secretion, and improve conception rate in the low-fertility females. Hormone treatments were administered to 38 parous control cows, 34 twice-open parous cows, and 27 twice-open nonparous heifers immediately after natural mating by a fertile bull. Blood samples were collected on Days 3, 6, 9, 12, and 18 after mating for determination of systemic progesterone concentrations. Pregnancy rate at necrospy approximately 33 days after mating (range 31–37) was higher in control cows (73.0%) than in twice-open cows (48.4%; P<0.05) or twice-open heifers (34.6%; P<0.01). Pregnancy rate was not affected by the HCG or GnRH treatment. The HCG treatment increased plasma progesterone concentrations in twice-open heifers but not in control or twice-open cows. Progesterone was unaffected by the GnRH treatment. Systemic progesterone concentrations were higher in control than in twice-open females but did not differ between pregnant and nonpregnant females of Days 3, 6, 9 and 12 after mating. Enhanced gonadotropin stimulation at estrus by injection of either HCG or GnRH did not increase pregnancy rate or systemic progesterone concentrations (except in HCG-treated twice-open heifers) in low- or high-fertility females. Lower pregnancy rates in twice-open females were not associated directly with the lower systemic progesterone concentrations.     

A field investigation of the effects of bovine viral diarrhea virus infection around the time of insemination on the reproductive performance of cattle

During a study of methods of synchronizing estrus in Bos indicus cattle, blood was collected from 169 heifers and 38 cows 2 to 3 days prior to artificial insemination (AI), and then again at Day 51 and Day 210 after AI to determine the incidence of infection with bovine viral diarrhea (BVD) virus. Prior to insemination 53 and 68% of the cows and heifers, respectively, were seronegative to the BVD virus. At Day 51 after AI, 70 and 32% of the seronegative cows and heifers, respectively, had seroconverted; but between Day 51 and Day 210, only 17 and 3% of the seronegative cows and heifers, respectively, had seroconverted. The Day- 51 pregnancy rate of cows which were immune (seropositive) to BVD virus infection at the time of AI was similar to the rate of the cows which became infected around the time of AI. However, the pregnancy rate of the immune heifers (44%, n=54) was significantly (P=0.04) greater than the rate of the heifers which became infected around the time of AI (24%, n=37). It was concluded that infection of susceptible females with BVD virus around the time of AI may significantly lower the pregnancy rate.     

Pregnancy rates in lactating dairy cattle following supplementation of progesterone after artificial insemination

Poor conception rates in highly productive lactating cattle is especially prevalent in large, intensively-managed commercial herds. One of the causative factors is sub-optimal pre-implantation embryonic development which appears to result from inadequate circulating concentrations of progesterone. In the present study, the efficacy of very modest progesterone supplementation, between Days 3.5 and 10 post-AI, on pregnancy rates was determined in a commercial herd where bovine somatotropin (bST) was used as a management tool. All lactating cattle that were deemed to be in estrus and inseminated over a 4-week period were randomly assigned to either a control group (no treatment) or CIDR-1.9g (previously used for estrous synchronization) treatment from Day 3.5 to Day 10 post-AI. Milk samples were collected four times: on the day of AI, at Day 2 or 3, at Day 4 and at Day 22 post-AI and were analyzed for progesterone content. Data from a total of 130 breedings were used in the final analysis. The CIDR treatment increased circulating concentrations of progesterone in treated animals over those of control animals on Day 4 by 0.7ng/ml (P<0.05) and increased pregnancy rate from 35% (22/63) to 48% (32/67) (P=0.068). The effect of treatment was greater in first and second lactation cows, where pregnancy rates were 33% (18/55) in controls and 51% (31/61) in treated animals (P=0.03). The results of this study indicate that the timing of onset of the progesterone influence is important for successful pregnancy outcome, particularly in first and second lactation cows.     

Use of short-term progestin treatment to resynchronize the second estrus following synchronized breeding in beef heifers

Two trials were conducted to evaluate the efficacy of short-term progestin administration to resynchronize the second estrus after artificial insemination in yearling beef heifers. In Trial 1 crossbred yearling heifers (n = 208) were synchronized with Syncro-Mate-B (SMB) and artificially inseminated (AI) between 48 and 54 h following implant removal. Implant removal is defined as Day 1. Following AI, the heifers were randomly assigned to 1 of 2 experimental groups. Group 1 heifers were fed melengestrol acetate (MGA) daily from Day 17 to 21 at a rate of 0.5 mg/head, while Group 2 control received no exogenous progestin during this period. Synchrony of estrus was defined as the 3-d period in which the highest number of heifers expressed behavioral estrus in each group. There was no difference (P < 0.05) in the pregnancy rate during the second estrus due to MGA supplementation. More MGA-treated heifers (P < 0.01) expressed estrus in a 3-d period than the controls. In Trial 2, yearling heifers (n = 108) were synchronized with 2 injections of PGF(2alpha) (second PGF(2alpha) injection is designated as Day 1) administered 14 d apart with AI 12 h after the onset of behavioral estrus. The heifers were then randomly assigned to 1 of the following 3 treatment groups after initial AI: 1) MGA fed at 0.5 mg/head daily from Days 17 to 21; 2) norgestomet administered in 6.0-mg implants from Days 17 to 21; 3) untreated control heifers. Blood samples were collected on Day 21 and analyzed for progesterone (P(4)). Elevated P(4) (> 1 ng/ml) on Day 21 indicated pregnancy to the first insemination. Synchrony among the 3 groups of heifers was similar (P > 0.10); however, the second estrus was less (P < 0.05) variable in the MGA and norgestomet treated heifers. During the resynchronized second estrus, conception rates were not affected by progestin treatment (MGA 40%, norgestomet 64%, and control 62%; P > 0.10). However, a proportion of heifers treated MGA 10% 4 36 and norgestomet 3% 1 36 expressed behavioral estrus during second estrus even though they were diagnosed as pregnant from first service by elevated P(4) levels on Day 21. We conclude that short-term use of progestin from Days 17 to 21 following AI causes closer synchrony of estrus; however, inseminating pregnant heifers that exhibit behavioral estrus may cause abortion.     

Luteotrophic influence of early bovine embryos and the relationship between plasma progesterone concentrations and embryo survival

This study was carried out to evaluate the luteotrophic influence of early (before Day 7 as well as after Day 7; Day 0=estrus) bovine embryos and the relationship between plasma progesterone (P4) concentrations and embryo survival. Virgin Holstein dairy heifers (n=325) from a single herd were randomly allocated to be nonbred, bred by artificial insemination (AI) or by embryo transfer (ET). Bred heifers were either treated with 1500 IU human chorionic gonadotrophin (hCG) on Day 7 of the estrous cycle or received no hCG treatment. Plasma P4 concentrations on Days 0, 5, 7, 10, 13, 15, 17, 19 and 21 were similar in pregnant AI- and ET-bred heifers and, this was observed in both hCG-treated and untreated females. Nonbred, AI- and ET-bred nonpregnant heifers (both hCG-treated and untreated) presented similar plasma P4 concentrations. Plasma P4 concentrations of pregnant heifers significantly deviated from those of nonpregnant and nonbred heifers on Day 17. In hCG-treated heifers, plasma P4 concentrations and Day 28 pregnancy rate were significantly higher in females with an induced accessory corpus luteum (CL) than in those females without an induced accessory CL. Treatment with hCG, although inducing the formation of accessory CL and significantly increasing plasma P4 concentrations had no significant effect on Day 28 pregnancy rate. In conclusion, this study does not support the existence of any peripherally detectable luteotrophic influence from early embryos (Days 5-7). Plasma P4 was only significantly related to embryo survival on Day 17, the time of expected onset of luteolysis.     

Effects of administration of gonadotropin-releasing hormone at artificial insemination on conception rates in dairy cows

A controlled trial investigating the effect on conception of administration of 250 μg of gonadotropin-releasing hormone (GnRH) at artificial insemination (AI) in dairy cows in seasonal or split calving herds was conducted. Time of detection of estrus, body condition, extent of estrous expression, treatment, breed, age and milk production from the most recent herd test of the current lactation was recorded. Cows were tested for pregnancy with fetal aging between 35 and 135 days after AI. Sixteen herds provided 2344 spring-calved cows and 3007 inseminations. Logistic regression adjusting for clustering at herd level was used to examine the effect of treatment for first (2344) and second (579) inseminations separately. For first AI, treatment significantly improved conception rate in cows with milk protein concentrations of 3.75% or greater and for cows with milk protein concentrations between 3.00% and 3.50% and less than 40 days calved; increased conception rate from 41.2% to 53.4%. Treatment reduced conception rates in cows with milk protein concentrations of 2.75% or less. Treating only cows identified as responding positively to treatment (11% of all study cows) was estimated to increase first service conception rate in herds from 48.1% to 49.4%. There was no significant effect of treatment on conception to second AI, nor any significant interactions. These findings indicate that GnRH at AI should be limited to the sub-group cows most likely to respond. The positive effect of GnRH at AI may be mediated through improved oocyte maturation and/or improved luteal function, rather than by reducing AI-to-ovulation intervals.     

Synchronization of estrus and fertility in beef cattle with two injections of buserelin and prostaglandin

Postpartum beef cows and heifers in Group 1 received 8 mug of buserelin on Day 0 (the beginning of the experiment) and 500 mug of cloprostenol (PGF) on Day 6 (GnRH I, n = 54). In Group 2 (GnRH II, n = 54), the females were injected with buserelin on Day 0 (8 mug) and Day 3 (4 mug), and PGF on Day 6 and Day 9 for females not detected in estrus previously. Animals were bred by AI 12 hours after the onset of estrus. Blood samples were collected on Day -11 and Day 0 to assess cyclicity and on Day 3 and Days 6 to 12 to examine luteal activity. Progesterone levels did not differ between the 2 groups between Days 0 to 9. In both groups, the proportion of spontaneous estruses from Days 0 to 6 was reduced. Precision of estrus was higher (P < 0.005) in the GnRH II group than in the GnRH I group of cows that were detected in estrus between Days 6 and 9. The synchronization rate, interval to estrus, pregnancy and conception rates were similar in GnRH I and GnRH II groups. The conception rate and interval to estrus were similar in cyclic and acyclic cows. Increasing the number of buserelin injections enhanced the precision of estrus, but not the conception rate, without any detrimental effect on luteal activity and induced more estruses in postpartum acyclic beef cattle.     

Effect of one or three timed artificial inseminations before natural service on reproductive performance of lactating dairy cows not observed for detection of estrus

The objectives were to determine the effects of one or three timed artificial insemination (AI) before natural service (NS) in lactating dairy cows not observed for detection of estrus on hazard of pregnancy, days nonpregnant, and 21-days cycle pregnancy rate. A total of 1050 lactating Holstein cows were subjected to a double Ovsynch program for their first postpartum AI. On the day of first AI (78 ± 3 days in milk), cows were blocked by parity and randomly assigned to receive either one timed AI (1TAI, n = 533) or three timed AI (3TAI, n = 517) before being exposed to NS. Cows assigned to 1TAI were exposed to bulls 7 days after the first AI. Nonpregnant cows in 3TAI were resynchronized with the Ovsynch protocol supplemented with progesterone twice, with intervals between AI of 42 days, before being exposed to NS 7 days after the third AI. Cows were evaluated for pregnancy 32 days after each timed AI, or every 28 days after being exposed to NS. Pregnant cows were re-examined for pregnancy 28 days later (i.e., 60-day gestation). Exposure to heat stress was categorized based on the first AI being performed during the hot or cool season, according to the temperature-humidity index. Body condition was scored at first AI. All cows were allowed a period of 231 days of breeding, after which nonpregnant cows were censored. Pregnancy to the first AI did not differ between 1TAI and 3TAI on Day 60 after insemination (30.8 vs. 33.5%). Cows receiving 3TAI had a 15% greater hazard of pregnancy and a 17% greater 21-days cycle pregnancy rate than 1TAI and these benefits originated from the first 84 days of breeding. These changes in rate of pregnancy reduced the median and mean days nonpregnant by 9 and 10 d, respectively. Despite the long inter-AI interval in cows subjected to 3TAI, reproductive performance was improved compared with a single timed AI and subsequent exposure to NS. In dairy herds that use a combination of AI and NS, allowing cows additional opportunities to AI before onset of breeding with bulls is expected to improve reproductive performance.     

Effects of abnormal ovarian cycles during pre-service period postpartum on subsequent reproductive performance of high-producing Holstein cows

The primary objective of this study was to investigate the effects of abnormal ovarian cycles during the pre-service postpartum period on subsequent reproductive performance of high-producing Holstein cows. The study was conducted in a commercial dairy farm with approximately 150 lactating cows, in a subtropical region of Japan. Animals were kept in free-stall barn, and fed a total mixed ration. Cows that calved from June 2001 to July 2002 were included in the study. Milk samples were collected twice weekly from 2 to 11 weeks postpartum, and progesterone concentrations in skim milk were determined by ELISA. After a voluntary waiting period of 40 days, cows detected in estrus were bred by artificial insemination (AI). Pregnancy was confirmed by palpation per rectum 40-70 days after AI. Out of a total of 91 cases, 39 (42.9%) had normal ovarian cycles (ovulation within 45 days after calving, followed by normal ovarian cycles), 32 (35.2%) had prolonged luteal phase (i.e. luteal activity for >20 days), and 12 (13.2%) had anovulation or delayed first ovulation (i.e. first ovulation did not occur until >45 days after calving). The remaining (8.8%) had other types of abnormalities. When compared with cows with a normal ovarian cycle, prolonged luteal phase cows had a lower 100 days AI submission rate, conception rate and pregnancy rate (84.2% versus 56.3%; P<0.05, 50% versus 16.7%; P<0.05 and 42.1% versus 9.4%; P<0.01, respectively), and longer intervals to first AI and to conception ( 67+/-6 days versus 98+/-7 days and 95+/-9 days versus 136+/-11 days; P<0.01 for each). Similarly, when compared with cows with normal ovarian cycles, those with anovulation had lower 100 days conception rate and pregnancy rate (50% versus 0%; P<0.05 and 42.1% versus 0%; P<0.01, respectively), and longer intervals to first AI and to conception ( 67+/-6 days versus 93+/-12 days; P<0.05 and 95+/-9 days versus 155+/-14 days; P<0.01, respectively). Survival analysis of the data for calving to conception interval showed that cows with prolonged luteal phase or anovulation were getting pregnant at a slower rate, and took longer to get pregnant than the cows with normal resumption of ovarian cyclicity postpartum. In conclusion, abnormal ovarian cycles during the pre-service period postpartum adversely affected reproductive performance, including AI submission rate, pregnancy rate, interval to first AI, and calving to conception interval in high-producing Holstein cows.     

Exposure of beef females to the biostimulatory effects of bulls with or without deposition of seminal plasma prior to AI

The objective of this experiment was to evaluate the biostimulatory effect of bull exposure, with or without the deposition of seminal plasma, on expression of estrus and pregnancy rate to AI in cattle. Beef heifers (n=86) and cows (n=193) were allocated to one of three treatments: (1) no bull exposure (CON; n=95), (2) exposure to a bull with a surgically-deviated penis for 21 d prior to AI (DB; n=88), or (3) exposure to a vasectomized bull for 21 d prior to AI (VB; n=96). The DB treatment provided the physical presence of a bull but prevented intromission, whereas the VB treatment allowed for intromission and deposition of seminal plasma but not spermatozoa. The estrous cycles of all females were synchronized using the Select Synch+CIDR protocol (GnRH+CIDR-7d-CIDR removal+PGF(2α), detection of estrus+AI 12h later for 84 h-clean-up TAI+GnRH). Pregnancy was detected via transrectal ultrasonography on d 35 post-AI. At the onset of the experiment, 75.7% of heifers and 86.1% of cows were estrous cycling. The percentages of females that displayed estrus were similar (P>0.05) among treatments (71.4%, 76.8%, and 74.4% for CON, DB, and VB, respectively). Pregnancy rates tended to be greater (P=0.06) in females in the DB treatment (60.5%) compared to females in the VB treatment (42.2%), with the control group intermediate (49.5%). In conclusion, biostimulation did not affect the expression of estrus but females exposed to the DB treatment tended to have an increased pregnancy rate.     

Concentrations of progesterone in milk as a monitor of early pregnancy diagnosis in dairy cows

The objectives of the experiment were to evaluate the efficacy of using progesterone concentrations in milk and palpation per rectum on days 21 or 22 postbreeding to estimate pregnancy and evaluate management practices; and to investigate physiological occurrences leading to incorrect diagnosis of pregnancy when serial samples of milk were collected. Of particular interest were indications of early embyronic death and insemination of cows not in estrus. Milk samples were collected at the afternoon milking of days 0 or 1 (day 0 = day of estrus), 9 or 10, 21 or 22 and 27 or 28 following breeding in 200 lactating dairy cows. Tentative diagnosis of pregnancy was made based on concentrations of progesterone in milk on days 21 and 22 alone and on days 21 or 22 and 27 or 28. In addition all cows were palpated per rectum on days 21 or 22 postbreeding and a tentative pregnancy diagnosis was made. Pregnancy was confirmed by examination of the genital tract per rectum between 35 and 50 days after breeding. Values of 4 ng/ml or greater and/or the presence of a mature corpus luteum were considered positive signs of pregnancy. Progesterone in milk ranged from 0.1 to 18 ng/ml. On days 0 or 1, 9 or 10, 21 or 22 and 27 or 28 concentrations of progesterone in milk averaged 1.5 +/- 0.3, 11.1 +/- 0.5, 12.0 +/- 0.4 12.5 +/- 0.5 ng/ml for pregnant cows. Corresponding samples from nonpregnant cows averaged 1.2 +/- 0.2, 10.3 +/- 0.4, 3.0 +/- 0.4, 6.8 +/- 0.6 ng/ml, respectively. Ninety-six and 104 cows were classified as pregnant and nonpregnant on days 21 or 22 as compared to 78 and 118 cows diagnosed as pregnant and nonpregnant on days 21 or 22 and 27 or 28 combined. Pregnancy detection by progesterone in milk on days 21 or 22 with pregnancy determined via rectal palpation 35 to 50 days postbreeding was 77 and 100% accurate for positive and negative diagnosis, respectively. The percent agreement using progesterone in milk on days 21 or 22 and 27 or 28 combined was 95 and 100%, respectively, for positive and negative diagnosis. Diagnosis based on rectal palpation 21 or 22 days postbreeding was 63 92 (69%) and 76 88 (87%) for pregnant and nonpregnant cows, respectively. Ten of the 200 cows had progesterone concentratins in milk of > 4 ng/ml at the time of breeding. Six of these cows were pregnant from a previous insemination. The other four cows were nonpregnant and were inseminated during the luteal phase of the cycle. In conclusion, measurement of progesterone in milk is a useful tool in early detection of pregnant and nonpregnant cows and may be useful in detecting reproductive problems in a dairy herd. It will probably be most useful when used in combination with later pregnancy diagnosis per rectum .