Modulation of expression of estrus, steroidogenesis and embryo development following peri-artificial insemination nutrient restriction in beef heifers.

Nutritional changes immediately after insemination cause increased embryonic mortality, but the mechanisms controlling this are not well known. Our objective was to evaluate the impact of nutritional change on estrus expression, steroid concentrations, peripheral and uterine luminal fluid metabolites, and embryo quality in beef heifers. Heifers (n = 139) were assigned to one of two pre-artificial insemination (AI) dietary treatments: LOW (≤ 90% NEm) or HIGH (≥ 139% NEm). Heifers were on treatment for 33-36 days before AI (d0) when half of the heifers in each treatment were randomly reassigned to generate four treatments; HIGH-HIGH, HIGH-LOW, LOW-HIGH, and LOW-LOW. Heifers remained on treatments until embryo collection (d 6-8). Negative energy balance was achieved among LOW heifers as demonstrated by body weight loss and increased NEFA concentrations (P < 0.05). Pre-AI treatment influenced expression of estrus (P = 0.05; HIGH 80.4 ± 4.0% vs. LOW 69.4 ± 4.2%). Estradiol concentrations and interval to estrus were not affected by treatment (P > 0.55); however, progesterone concentrations were reduced among LOW compared to HIGH (3.57 ± 0.27, 4.64 ± 0.26 ng/mL, respectively; P = 0.004), and heifers maintained on the HIGH pre-AI diet had consistently greater concentrations of progesterone from d 0 to d 8 (P = 0.014). Pre-AI treatment influenced embryo stage (P = 0.05; HIGH 3.61 ± 0.32 vs. LOW 2.72 ± 0.30). Post-AI treatment affected embryo grade (P = 0.02; HIGH 1.78 ± 0.23 vs. LOW 2.64 ± 0.27). In summary, pre-AI nutrient restriction caused decreased expression of estrus, reduced progesterone concentrations after AI, and negatively impacted embryo development, while post-AI restriction hindered embryo quality.

Effect of progesterone supplementation in a resynchronization protocol on follicular dynamics and pregnancy success.

The objective of this study was to evaluate the necessity of a controlled internal drug releasing device (CIDR) in a fixed-time AI resynchronization protocol as well as to compare a commercially available blood pregnancy test with transrectal ultrasonography for Day 28 pregnancy detection. Over a two-year period, beef cows and heifers from twelve herds were inseminated using the 7-day CO-Synch + CIDR protocol. On Day 21 following the first insemination, the protocol was repeated, with animals receiving either a CIDR or no CIDR. Pregnancy status (AI1) was determined on Day 28 by both transrectal ultrasonography and the IDEXX Rapid Visual Pregnancy Test. Non-pregnant animals by both methods (CIDR: n = 190 cows, n = 228 heifers; no CIDR: n = 185 cows, n = 223 heifers) received an injection of Prostaglandin F2alpha and were inseminated at the appropriate time or bred following detection of estrus. Corpora lutea (CL) number and largest follicle diameter were recorded on a subset of non-pregnant animals (CIDR: n = 66 cows, n = 46 heifers; no CIDR: n = 76 cows, n = 41 heifers) at time of pregnancy diagnosis on Day 28. Final pregnancy status was determined a minimum of 31 days following the second AI (AI2). The GLIMMIX procedure of SAS was utilized for estrus and pregnancy data; while the MIXED procedure was utilized for analyses of CL number and largest follicle diameter. There was no effect (P ≥ 0.55) of treatment on AI1 pregnancy, AI2 pregnancy, or overall pregnancy rates. The presence of a CIDR during the resynchronization increased (P < 0.001) estrus expression prior to AI2. There was an effect of treatment by age on AI2 pregnancy (P < 0.01); heifers that received a CIDR had greater AI2 pregnancy rates than heifers that did not receive a CIDR (P = 0.04), but there was no difference between cows with and without a CIDR. Treatment had no effect (P > 0.10) on embryonic loss (between the first and second pregnancy diagnosis), CL number, or follicle diameter. Although, there was a tendency for the interaction of treatment by age on follicle size (P = 0.07), with cows having larger follicles than heifers in the no CIDR group but not the CIDR group. In conclusion, use of a CIDR in this resynchronization protocol increased estrus expression, increased AI2 pregnancy for heifers, but did not improve pregnancies in cows, and did not influence overall pregnancy or embryonic loss.

Role of preovulatory concentrations of estradiol on timing of conception and regulation of the uterine environment in beef cattle.

The ability to induce ovulation with an injection of GnRH facilitated the development of fixed-time artificial insemination (AI) protocols. However, animals that exhibit estrus prior to fixed-time AI have greater pregnancy success. Thus, the objectives of the present experiments were to determine the impact of estrus expression prior to fixed-time AI on timing of conception and to characterize the role of preovulatory estradiol in regulating changes in expression of uterine genes. In experiment 1, data were collected from 4,499 beef cows inseminated by fixed-time AI in 31 different herds. Animals that did not conceive to AI but exhibited estrus before timed-AI were more likely to conceive during cycle 1 after AI, and overall conceived earlier in the breeding season compared to animals that did not exhibit estrus. In experiment 2, beef cows were synchronized using a fixed-time AI protocol. Uterine horn biopsies and blood samples were collected on Day 0, 5, 10, or 16. Concentrations of estradiol on Day 0 did not influence expression of progesterone receptor, ER beta, or oxytocin receptor. Increased concentrations of estradiol on Day 0 increased expression of ER alpha from Days 0 to 5 of the estrous cycle. Furthermore, cows with increased concentrations of estradiol on Day 0 had increased expression of inhibin beta A, and uterine milk protein precursor. Thus, animals that do not exhibit estrus prior to fixed-time AI had decreased breeding season pregnancy success and conceived later in the breeding season, and preovulatory concentrations of estradiol likely play a major role in this establishment of pregnancy, not only directly by regulating uterine gene expression, but also indirectly throughout the subsequent estrous cycle.

Influence of Sexcel™ (gender ablation technology) gender-ablated semen in fixed-time artificial insemination of beef cows and heifers.

Previous research has indicated that gender-sorted semen has decreased conception rates compared to conventional semen. A new method to skew the gender ratio of semen has been developed that does not use traditional sorting technology. Thus, the objective of this study was to evaluate the use of gender-ablated semen in a fixed-time AI protocol. Beef heifers and cows (n = 878) from six herds were synchronized with the 7-d CO-Synch plus CIDR protocol, and artificially inseminated (AI) after CIDR removal (cows 60-66 h; heifers 52-56 h). Estrus detection aids were applied at CIDR removal and estrus activity was determined at time of AI. Animals remained separated from bulls for at least 10 d after AI. Pregnancy success and fetal age were determined between d 28 and 70 after AI. Semen from five sires was utilized in this study, with two sires used in each herd. Sire 1 was used in all herds and the use of the second sire varied by herd (both conventional and gender-ablated semen from each sire was used in each herd). Data were analyzed using the GLIMMIX procedure in SAS and included the influence of semen type, estrus expression, semen type by estrus expression, dam age, and sire in the model. Herd was included as a random variable. Overall, conventional semen had greater conception rates compared to gender-ablated semen (P < 0.01; 67% vs 52%), and cows that had fully activated patches and partially activated patches had greater conception rates compared to animals that had not exhibited estrus (P < 0.01; 69%, 65%, and 45%). There was no semen type by estrus expression interaction (P = 0.24). Conception rates tended to be greater for conventional semen among animals that had activated patches (P = 0.06; 73% vs 65%) and partially activated patches (P = 0.06; 72% vs 59%). Conception rates were greater for conventional semen (P < 0.01; 56% vs 33%) among animals that did not exhibit estrus. There was no effect of dam age (P = 0.40) or sire (P = 0.92) on conception rates. In conclusion, Sexcel™ gender-ablated semen can successfully be used in fixed-time AI protocols among animals that exhibit estrus, but caution should be used among animals that do not exhibit estrus.

Chemical and toxicologic assessment of organic contaminants in surface water using passive samplers.

Passive sampling methodologies were used to conduct a chemical and toxicologic assessment of organic contaminants in the surface waters of three geographically distinct agricultural watersheds. A selection of current-use agrochemicals and persistent organic pollutants, including polycyclic aromatic hydrocarbons, polychlorinated biphenyls, and organochlorine pesticides, were targeted using the polar organic chemical integrative sampler (POCIS) and the semipermeable membrane device passive samplers. In addition to the chemical analysis, the Microtox assay for acute toxicity and the yeast estrogen screen (YES) were conducted as potential assessment tools in combination with the passive samplers. During the spring of 2004, the passive samplers were deployed for 29 to 65 d at Leary Weber Ditch, IN; Morgan Creek, MD; and DR2 Drain, WA. Chemical analysis of the sampler extracts identified the agrochemicals predominantly used in those areas, including atrazine, simazine, acetochlor, and metolachlor. Other chemicals identified included deethylatrazine and deisopropylatrazine, trifluralin, fluoranthene, pyrene, cis- and trans-nonachlor, and pentachloroanisole. Screening using Microtox resulted in no acutely toxic samples. POCIS samples screened by the YES assay failed to elicit a positive estrogenic response.