G. C. Lamb

Fixed-time artificial insemination with estradiol and progesterone for Bos indicus cows I: Basis for development of protocols

Five experiments were conducted on commercial farms in Brazil aiming to develop a fixed-time artificial insemination (TAI) protocol that achieved pregnancy rates between 40% and 55% in Bos indicus cows. These studies resulted in the development of the following protocol: insertion of an intravaginal device containing 1.9 g of progesterone (CIDR) plus 2.0mg im estradiol benzoate on Day 0; 12.5mg im dinoprost tromethamine on Day 7 in cycling cows or on Day 9 in anestrous cows; CIDR withdrawal plus 0.5mg im estradiol cypionate plus temporary calf removal on Day 9; TAI (48h after CIDR withdrawal) plus reuniting of calves with their dams on Day 11. Reduced dose of prostaglandin F(2alpha) (PGF(2alpha); 12.5mg im dinoprost tromethamine) effectively caused luteolysis. In cycling cows, fertility was greater when the treatment with PGF(2alpha) was administered on Day 7 than on Day 9, but in anestrous cows, no effects of time of the PGF(2alpha) treatment were found. Estradiol cypionate effectively replaced estradiol benzoate or gonadotropin-releasing hormone as the ovulatory stimulus, reducing labor and cost. In this protocol, CIDR inserts were successfully used four times (9 d each use) with no detrimental effects on fertility.

Fixed-time artificial insemination with estradiol and progesterone for Bos indicus cows II: strategies and factors affecting fertility.

In Experiments 1, 2, and 3, we evaluated the effects of temporary weaning (TW), equine chorionic gonadotropin (eCG), and follicle-stimulating hormone (FSH) treatments on results of a fixed-time artificial insemination (TAI) protocol in postpartum Bos indicus cows. In Experiment 1, treatment with 400 IU eCG or with TW for 48 h consistently improved pregnancy rates (PRs) at TAI, but, in Experiment 2, FSH treatment was less effective than eCG or TW. In Experiment 3, the inclusion of eCG treatment in cows subjected to TW did not improve PRs. We concluded that TW or 400 IU eCG should be included in the TAI protocol in postpartum Bos indicus cows to enhance fertility. In Experiment 4, we used records from heifers and cows treated with the proposed protocol during the 2006-2007 (n=27,195) and 2007-2008 (n=36,838) breeding seasons from multiple locations in Brazil to evaluate factors potentially affecting PRs. Overall PR at TAI was 49.6% (31,786 of 64,033). Pregnancy rate differed (P<0.01) among farm within location (results ranging between 26.8% and 68.0%; P<0.01), cow group within farm, by breed (Bos indicus, 48.3% [26,123 of 54,145]; Bos taurus, 61.7% [3652 of 5922]; and crossbred Bos indicus x Bos taurus, 50.7% [2011 of 3966]), category (nulliparous, 39.6% [2095 of 5290]; suckled primiparous, 45.2% [3924 of 8677]; suckled multiparous, 51.8% [24,245 of 46,767]; and nonsuckled multiparous, 46.1% [1522 of 3299]), body condition score at TAI (< or =2.5, 43.0% [3409 of 7923]; 3.0, 49.6% [18,958 of 38,229]; and > or =3.5, 52.7% [9419 of 17,881]). Days postpartum at beginning of protocol did not affect PR (30 to 60 d, 47.6% [4228 of 8881]; 61 to 90 d, 51.7% [16,325 to 31,572]; and 91 to 150 d, 50.8% [7616 to 14,991]; P>0.1). Pregnancy rate was also consistently affected (P<0.01) by sire (results ranging from 7.2% to 77.3%) and artificial insemination technician (results ranging from 15.1% to 81.8%).

Evaluating recipient and embryo factors that affect pregnancy rates of embryo transfer in beef cattle

The objectives of this experiment were to determine the effects of corpus luteum characteristics, progesterone concentration, donor-recipient synchrony, embryo quality, type, and developmental stage on pregnancy rates after embryo transfer. We synchronized 763 potential recipients for estrus using one of two synchronization protocols: two doses of PGF2alpha (25 mg i.m.) given 11 d apart (Location 1); and, a single norgestomet implant for 7 d with one dose of PGF2alpha (25 mg i.m.) 24 h before implant removal (Location 2). At embryo transfer, ovaries were examined by rectal palpation and ultrasonography. Of the 526 recipients presented for embryo transfer, 122 received a fresh embryo and 326 received a frozen embryo. Pregnancy rates were greater (P < 0.05) with fresh embryos (83%) than frozen-thawed embryos (69%). Pregnancy rates were not affected by embryo grade, embryo stage, donor-recipient synchrony, or the palpated integrity of the CL. Corpus luteum diameter and luteal tissue volume increased as days post-estrus for the recipients increased. However, pregnancy rates did not differ among recipients receiving embryos 6.5 to 8.5 days after estrus (P > 0.1). There was a significant, positive simple correlation between CL diameter or luteal tissue volume and plasma progesterone concentration (r = 0.15, P < 0.01 and r = 0.18, P < 0.01, respectively). There were no significant differences in mean CL diameter, luteal volume or plasma progesterone concentration among recipients that did or did not become pregnant after embryo transfer. We conclude that suitability of a potential embryo transfer recipient is determined by observed estrus and a palpable corpus luteum, regardless of size or quality.

Detection of anovulation by heatmount detectors and transrectal ultrasonography before treatment with progesterone in a timed insemination protocol

Our objective was to determine the accuracy of identifying noncycling lactating dairy cows before the application of a timed artificial insemination (AI) protocol [with or without progesterone supplementation via a controlled internal drug-release (CIDR) insert and 2 different timings of AI] by using heatmount detectors and a single ovarian ultrasound examination. At 6 locations in the Midwest, 1,072 cows were enrolled in a Presynch protocol (2 injections of PGF(2alpha) 14 d apart), with the second injection administered 14 d before initiating the Ovsynch protocol (injection of GnRH 7 d before and 48 h after PGF(2alpha) injection, with timed AI at 0 or 24 h after the second GnRH injection). Heatmount detectors were applied to cows just before the first Presynch injection, assessed 14 d later at the second Presynch injection (replaced when activated or missing), and reassessed at initiation of the Ovsynch protocol. Ovaries were examined for the presence of a corpus luteum (CL) by ultrasound before the initiation of treatment. Treatments were assigned to cows based on the presence or absence of a CL detected by ultrasound: 1) no CL + no CIDR; 2) no CL + CIDR insert for 7 d; and 3) CL present. Further, alternate cows within the 3 treatments were assigned to be inseminated concurrent with the second GnRH injection of Ovsynch (0 h) or 24 h later. Pregnancy was diagnosed at 33 and 61 d after the second GnRH injection. By using low (<1 ng/mL) concentrations of progesterone in serum as the standard for noncycling status, heatmount detectors were activated on a large percentage of noncycling cows (>60%), whereas the single ultrasound examination incorrectly classified noncycling cows only 21% of the time. Conversely, cycling cows (progesterone > or =1 ng/mL) were correctly identified 70 to 78% of the time by heatmount detectors, but 85 to 92% were correctly identified by ultrasound. Overall accuracy of heatmount detectors and ultrasound was 71 and 84%, respectively. Application of progesterone to cows without a CL at the time of the first injection of GnRH reduced the incidence of ovulation but increased the proportions of pregnancies per AI at d 33 or 61 compared with nontreated cows without a CL at the onset of the Ovsynch protocol. Percentages of cows pregnant and pregnancy survival did not differ for cows having a CL before treatment compared with those not having a CL and treated with progesterone. Compared with no response, when a follicle ovulated in response to the first GnRH injection, percentage of cows becoming pregnant after the timed AI increased from 33.3 to 41.6%. Timing of AI at 0 or 24 h after the second GnRH injection did not alter pregnancies per AI, but cows having luteal activity before treatment had improved pregnancies per AI compared with noncycling cows. We conclude that identifying noncycling cows by ultrasound was more accurate than by heatmount detectors. Subsequent progesterone treatment of previously cycling cows not having a CL at the onset of Ovsynch increased the proportion of pregnant cows, equal to that of cows having a CL but not treated with progesterone.

Effects of acclimation to human interaction on performance, temperament, physiological responses, and pregnancy rates of Brahman-crossbred cows.

The objective of this study was to evaluate, over 2 consecutive years, the effects of acclimation to human interaction on performance, temperament, plasma concentrations of hormones and metabolites, and pregnancy rates of Brahman-crossbred cows. A total of 160 Braford and 235 Brahman x British cows were assigned to the 2-yr study. Approximately 45 d after weaning (August 2006) in yr 1, cows were evaluated for BW, BCS, and temperament (chute score, pen score, and exit velocity), stratified by these measurements in addition to breed and age, and randomly allocated to 14 groups (Braford = 8; Brahman x British = 6). Groups were randomly assigned to the control or acclimation treatment. In yr 2, cows were reevaluated within 45 d after weaning (August 2007) for BW, BCS, and temperament, stratified, and divided into 14 groups similarly as in yr 1, but in such a way that cows received the same treatment assigned in yr 1. Cows were acclimated to human interaction from August to January, and the acclimation process consisted of the same person visiting groups twice weekly and offering approximately 0.05 kg of range cubes per cow (as-fed basis). In January of both years, cow temperament, BW, and BCS were reassessed and cows were exposed to a 90-d breeding season. Blood samples were collected at the beginning of the acclimation period (August) and breeding season (January) for determination of plasma cortisol, IGF-I, and acute phase proteins. A treatment x breed interaction was detected during yr 1 (P < 0.01) for pregnancy analysis because acclimated Braford cows conceived earlier and at a greater percentage (P < 0.01) compared with control cows. According to values obtained at the beginning of breeding and pooled across treatments and breeds, IGF-I concentrations and BCS affected quadratically (P < 0.05), and concentrations of ceruloplasmin and haptoglobin decreased linearly (P < 0.05), the probability of pregnancy during both years. Temperament and cortisol concentrations decreased the probability of pregnancy linearly (P < 0.05) during yr 1 (only Braford cows for cortisol analysis) and affected the probability of pregnancy quadratically (P < 0.05) during yr 2. Results from this study indicate that acclimation did not affect cow temperament and physiological responses but did increase pregnancy rates of Braford cows during yr 1. Further, measurements and physiological responses associated with temperament influenced the probability of cows becoming pregnant during the breeding season.

Control of the estrous cycle to improve fertility for fixed-time artificial insemination in beef cattle: a review.

Early estrus-synchronization protocols focused on regressing the corpus luteum (CL) with an injection of PGF(2alpha) followed by detection of estrus or involved the use of exogenous progestins that prevent estrus from occurring. Later, protocols combining the use of PGF(2alpha) and exogenous progestins were developed. Gonadotropin-releasing hormone was utilized to control follicular waves, synchronize ovulation, or to luteinize large dominant follicles. Our research aimed to develop reliable protocols that 1) relied solely on fixed-timed AI (TAI); 2) required a maximum of 3 animal handlings, and 3) were successful in estrous-cycling and noncycling females. In cows, insertion of an intravaginal progesterone insert during the 7-d interval between the initial GnRH and PGF(2alpha) injections enhanced pregnancy rates by 9 to 10%. In a multi-location study, a TAI protocol yielded pregnancy rates similar to a protocol involving detection of estrus plus a fixed-time clean-up AI for females not detected in estrus (54 vs. 58%, respectively, for cows and 53 vs. 57%, respectively, for heifers). Initiation of estrous cycles in noncycling cows is likely the primary manner in which beef producers may improve fertility in response to estrus synchronization and TAI protocols. Treatment of noncycling females with progesterone and GnRH increases the percentage of cycling females and improves fertility to a TAI, but inducing cyclicity with hCG failed to enhance fertility in TAI protocols. Supplementing progesterone after TAI failed to increase pregnancy rates in beef cattle. In contrast, administration of hCG 7 d after TAI induced an accessory CL, increased progesterone, and tended to enhance pregnancy rates. Development of TAI protocols that reduce the hassle factors associated with ovulation synchronization and AI provide cattle producers efficient and effective tools for capturing selective genetic traits of economic consequences. Location variables, however, which may include differences in pasture and diet, breed composition, body condition, postpartum interval, climate, and geographic location, affect the success of TAI protocols.

Prolonging the MGA-prostaglandin F2α interval from 17 to 19 days in an estrus synchronization system for heifers.

Our objective was to determine whether extending the interval from 17 to 19 d between removal of melengestrol acetate (MGA) feed and administration of PGF2 alpha would alter conception rates, pregnancy rates and the degree of synchrony in replacement beef heifers. A commercial heifer operation in north-central Kansas purchased 591 Angus x Hereford heifers from 12 sources. Prior to the spring breeding season, 14% of the heifers were culled. The remaining heifers were assigned randomly to 2 MGA-PGF2 alpha synchronization systems. All heifers were fed MGA (0.5 mg/head/d) for 14 d, and PGF2 alpha was administered either 17 or 19 d after the completion of MGA feeding. Heifers were inseminated artificially for 30 d followed by 30 d of natural mating. Based on each source, first-service conception rates ranged from 66 to 90%, whereas overall pregnancy rates ranged from 91 to 100%. Heifers given PGF2 alpha on Day 17 after MGA had first-service conception rates of 75.9% compared with 81.4% for heifers receiving PGF2 alpha on Day 19. In response to the PGF2 alpha injection, 99% of the Day 19 heifers that were detected in estrus were inseminated artificially by 72 h after the PGF2 alpha injection, whereas 74% of the heifers in the Day 17 treatment were inseminated by that time. Average interval to artificial insemination (AI) after PGF2 alpha was greater (P < 0.01) for the Day 17 heifers (73.1 +/- 1.1 h) than for the Day 19 heifers (56.2 +/- 1.1 h). No differences in conception rates or overall pregnancy rates occurred; however, heifers receiving PGF2 alpha on Day 19 after MGA had shorter intervals to estrus, and a greater proportion was inseminated within 72 h after PGF2 alpha, thus possibly facilitating successful timed insemination of the remaining heifers not yet inseminated by that time.

Postpartum endometrial cytology in beef cows.

The objectives were to characterize postpartum endometrial cytology and to determine the prevalence of subclinical endometrial inflammation and its impact on reproduction in beef cows. Samples for endometrial cytology (low-volume uterine lavage) were obtained from 135 of 137 Angus cows (2-87d postpartum) in northern Minnesota, 26d before breeding started. Agreement between examiners for subjective inflammation scores was very high (kappa=0.971); the correlation between these scores and PMN counts was high (r=0.83; P<0.001), validating subjective categorization. The proportion of PMN and large mononuclear cells (principally macrophages) declined with postpartum interval (P<0.001), whereas small mononuclear cells were consistently present (and not significantly affected by postpartum interval). Pregnancy rate to fixed-time AI was 29% and overall pregnancy rate was 89%. There was no association between cell type and ultimate pregnancy status or day of conception (P>0.10). Although inflammation later in the postpartum period apparently impaired subsequent reproduction in dairy cows, in cows >50 d postpartum at sample collection in the present study, no cytological parameter significantly predicted final pregnancy status or day of conception. Previous twinning increased the risk of subclinical endometritis (P=0.02), but not the probability of becoming pregnant (P=0.14). In conclusion, we inferred that beef cows had the ability to clear uterine inflammation after resumption of ovarian cyclicity.

Effects of supplementation frequency on performance, reproductive, and metabolic responses of Brahman-crossbred females

Two experiments were conducted to compare performance and metabolic responses of beef females consuming low-quality forages and offered an energy supplement based on fibrous byproducts daily (S7) or 3 times per week (S3) at similar weekly rates. In Exp. 1, BW gain, reproductive performance, mRNA expression of hepatic and skeletal muscle genes associated with nutritional metabolism and growth, and concentrations of blood urea nitrogen (BUN), plasma glucose, insulin, and IGF-I were assessed in 56 Brahman x Angus heifers supplemented at a daily rate of 1.0% of BW. Mean BW gain was greater (P = 0.03) for S7 compared with S3 heifers. Treatment x sampling day interactions were detected (P < 0.01) for all blood measurements. Heifers provided S7 had less daily variation in concentrations of BUN, glucose, and insulin, and frequently had greater (P < 0.05) concentrations of IGF-I compared with S3 heifers. Expression of liver IGF-I mRNA was greater (P = 0.04) for S7 heifers compared with S3 heifers. Treatment x day interactions were detected (P </= 0.05) for mRNA expression of liver IGFBP-3, gluconeogenic enzymes, and muscle myostatin because the expression of these transcripts was greater (P < 0.05) for S3 heifers when both treatment groups were supplemented, but was similar or greater (cytosolic phosphoenolpyruvate carboxykinase; P = 0.04) for S7 heifers when only these were supplemented. Attainment of puberty and pregnancy were hastened (P = 0.03 and 0.02, respectively) in S7 heifers compared with S3 heifers. In Exp. 2, 12 Brahman x British mature cows received S3 or S7 for a 3-wk period at a daily rate of 0.5% of BW. Concentrations of BUN were greater for S7 compared with S3 cows (P < 0.03). A treatment x time interaction was detected (P = 0.01) for insulin concentrations because a time effect was significant (P < 0.01) for S3 but not S7 cows. With the advance of the experiment, concentrations of IGF-I increased for S7 (P < 0.01) but not S3 cows (treatment x week interaction; P = 0.02). The combined expression of gluconeogenic enzymes mRNA tended to be greater (P = 0.09) for S3 cows when both treatment groups received supplements, but was greater (P = 0.03) for S7 cows when only these were supplemented (treatment x day interaction; P < 0.01). In conclusion, offering an energy supplement based on fibrous byproducts daily instead of 3 times weekly enhanced the nutritional and metabolic status of forage-fed Brahman-crossbred females, resulting in improved growth and reproductive performance of developing heifers.

Effects of vaccination on the acute-phase protein response and measures of performance in growing beef calves

Two experiments were conducted to evaluate the influence of vaccination on the acute-phase protein (APP) reaction (Exp. 1 and 2) and measures of performance (Exp. 2) in growing beef calves. In Exp. 1, the APP reaction was assessed in newly weaned steers administered 1 of 3 treatments (n = 8 steers/treatment), consisting of 1) Mannheimia haemolytica vaccine (One Shot; Pfizer Inc., New York, NY), 2) Clostridium vaccine (UltraBac 7; Pfizer, Inc.), or 3) saline-injected control. Blood samples for the evaluation of APP concentrations were collected on d 0, 1, 3, 5, 7, 10, and 14 and steer BW measured on d 0 and 21 relative to treatment administration. Plasma concentrations of haptoglobin (Hp) increased (P < 0.05) in vaccinated but not control calves and reached a peak on d 3 and 5 for steers receiving Mannheimia haemolytica and Clostridium vaccine, respectively. Plasma concentrations of ceruloplasmin (Cp) and fibrinogen (Fb) increased (P < 0.05) in all calves after treatment administration and Fb concentrations were greatest (P < 0.01) in calves receiving Mannheimia haemolytica vaccine on d 3 and 5 compared with the other treatments. There were no treatment effects (P = 0.44) on 21-d steer ADG (0.43 kg/d; SEM = 0.082). In Exp. 2, 23 heifers were randomly assigned to 2 treatments: 1) vaccinated (Mannheimia haemolytica vaccine (One Shot; n = 12) and 2) saline control (n = 11). After vaccination, blood samples were collected for determination of APP concentrations on d 0, 3, 6, 9, 12, and 15. During this period, individual heifer DMI was measured using an automated feed intake measuring system (Model 4000E; GrowSafe Systems Ltd., Airdrie, Alberta, Canada). Initial and final shrunk BW did not differ (P > 0.36) among treatments. On d 1, plasma Cp concentrations increased (P < 0.01) sharply in vaccinated heifers but not control heifers and were greater (P < 0.05) in vaccinated vs. control heifers on d 3, 6, 9, and 12 relative to injection. Daily DMI did not differ (P = 0.66) among treatments (average = 9.1 kg/d; SEM = 0.34); however, ADG and G:F were greater (P ≤ 0.05) for control vs. vaccinated heifers (1.14 vs. 0.87 kg/d and 0.13 and 0.10 kg, respectively; SEM = 0.064 and 0.011). These data indicate that within a 2 wk period after vaccination, beef calves experience an acute-phase protein response, which may result in reduced ADG and feed efficiency.