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The microsatellites HEL5, HEL9, INRA063, and BM2113 were used to analyze genetic similarities and differences of geographically isolated Criollo cattle herds in Mexico. Criollo cattle from five counties within the state of Chihuahua and one county from the state of Tamaulipas (n = 60) were sampled. The five counties in Chihuahua included Cerocahui (n = 14), Chinipas (n = 10), Guachochi (n = 15), Morelos (n = 30), and Temoris (n = 9). Samples of DNA were amplified by PCR and separated on a 7% polyacrylamide gel. Microsatellite size was established by comparison to M13mp18 DNA ladder and a documented set of four bovine controls. Allele frequencies and genotypic deviations from Hardy-Weinberg equilibrium were tested using the GENEPOP program. Eleven alleles were generated at HEL5 for the populations sampled (149 to 169 bp). Allele frequencies were greatest for the 163-bp allele in Criollo cattle from Cerocahui, Chinipas, Moralos, and Tamaulipas (0.23 to 0.5). Cattle from Guachochi had an allele frequency of 0.38 for the 151-bp allele, and cattle from Temoris had an allele frequency of 0.25 for the 149- and 167-bp alleles, with no 163-bp allele. Amplification with HEL9 produced 12 alleles (145, 149 to 169 bp) and showed common high-frequency alleles at 149, 157, and 159 bp for animals from all regions. The Chinipas population showed a moderate allele frequency at 145 bp; no other regions contained this allele. For INRA063 there were five alleles with 182 and 184 bp in low frequency. For BM2113 there were 10 alleles in the Criollo cattle (125 to 143 bp), with an equal distribution of frequencies for all alleles. In two regions, Guachochi and Morelos, genotypic frequencies deviated from Hardy-Weinberg equilibrium. Cattle from the Temoris region were genetically most distant from Criollo cattle of the other five regions. Key Words: Genetic Analysis, Genetic Variation, Microsatellites

Contents The effects of supplemented protein level (PL) during the periconceptional period and their interaction with body condition were evaluated in sheep. Multiparous Rambouillet ewes (n = 12) received two PL of rumen undegradable protein (UIP) during a 30-day pre-mating and 15-day post-mating period: low [LPL, 24% crude protein (CP), 14 g UIP and 36 g/CP animal/day] and high [HPL, 44% CP, 30 g UIP and 50 g/CP animal/day]. While ovulation rate (OR) did not differ between treatments (1.6 ± 0.5, mean ± SEM), a lower fertility rate, a decreased embryo number and a reduced uterine pH (UpH) was observed in the HPL group (p < 0.05), irrespective of BC. Luteal tissue weight, volume and progesterone secretion did not differ among treatments. Sheep with lower UpH also had lower conceptus weight (Cwt; p < 0.05, r = 0.65) and conceptuses with lower mass tended to secrete less INF-τ and IGF-1, and the correspondent endometrial explants had a higher basal PGF2α release. Current study indicates that high protein diets during the periconceptional period in sheep modify uterine and embryonic relationships, increasing early pregnancy losses and inducing embryo growth retardation. Surviving embryos were affected by weight reductions, which could compromise later foetal growth and birth weight. Results evidence the key role of a balanced diet in reproductive success and indicate that the quality and nutrient composition of the maternal diet are essential for an adequate establishment of pregnancy, having paramount effects on the interplay of the embryo and the uterus. [ABSTRACT FROM AUTHOR]

Consistent responses of young postpartum beef cows to protein supplementation could improve reproduction and cow sustainability. A 2-yr study conducted at the Corona Range and Livestock Research Center from February to July in 2003 (n = 51) and 2004 (n = 40) evaluated responses of 2- and 3-yr-old postpartum beef cows grazing dormant native range to three protein supplements with increasing glucogenic potential (GP). Supplements were fed at 1135 g•cow-1•d-1 twice weekly for approximately 70 d and provided 1) 341 g CP, 142 g ruminally undegradable protein (RUP), 57 g GP (RUP0), 2) 341 g CP, 151 g RUP + 80 g propionate salt (NutroCAL™, Kemin Industries, Inc.), 124 g GP (RUP80), or 3) 341 g CP, 159 g RUP + 160 g propionate salt, 192 g GP (RUP160). Weekly serum samples were composited and analyzed for glucose, non-esterified fatty acids (NEFA), and serum urea nitrogen (SUN). Supplement × yr interactions were observed for each metabolite (P ≤ 0.04). Cows fed RUP0 had higher glucose concentrations in 2004 than in 2003. In 2003, cows fed RUP160 had higher serum SUN concentrations than RUP80-fed cows; the opposite effect was observed in 2004. However, SUN concentrations for all cows were higher in 2004 than in 2003. Serum NEFA concentrations of RUP160-fed cows were higher in 2003 than in 2004. Cows fed RUP80 had the most consistent response, regardless of yr. Implications of this study suggest that cows fed the moderate level of GP were able to compensate for yr differences. [ABSTRACT FROM AUTHOR]

The objective was to evaluate the effects of GnRH or PGF2α-based synchronization and resynchronization programs on fertility in lactating dairy cows. For experiment 1, cows (n = 1,521) were presynchronized with 2 injections of PGF given at 36 and 50 DIM and assigned to 1 of 3 protocols: Ovsynch [OVS; n = 552; GnRH injection, PGF2α injection 7 d later, GnRH injection 56 h later, and timed artificial insemination (TAI) 16 h later] beginning at 14 d after presynchronization (PS), GnRH-GnRH-PGF2α-GnRH (GGPG; n = 402) treatment with a GnRH injection given 7 d after PS and OVS 7 d later, or PGF2α-GnRH-PGF2α-GnRH (P7GPG; n = 567) treatment with a PGF2α injection given 7 d after PS and OVS beginning 7 d later. Experiment 2 cows (n = 2,327) were assigned to 1 of 3 resynchronization protocols 7 d before nonpregnancy diagnosis (NPD): GGPG cows (n = 458) received a GnRH injection at enrollment and OVS at NPD; P7GPG cows (n = 940) received a PGF2α injection at NPD and OVS 7 d later; and P11GPG cows (n = 929) received a PGF2α injection 3 d after NPD and OVS 11 d later. In both experiments, cows were artificially inseminated upon estrus detection (ED). In experiment 1, 52.3% of cows were artificially inseminated upon ED, with GGPG having reduced ED (GGPG = 46.8 vs. OVS = 50.7 and P7GPG = 57.7%). Treatments did not affect overall pregnancy per artificial insemination (P/AI) at 36 and 66 d after AI (OVS = 34.1 and 32.3, P7GPG = 34.6 and 31.9, and GGPG = 31.3 and 28.1%, respectively) or pregnancy loss but cows artificially inseminated upon ED had higher P/AI than cows undergoing TAI (ED cows = 37.9 vs. TAI cows = 28.8%). Treatment did not affect P/AI for cows artificially inseminated upon ED or TAI at 36 and 66 d after AI (OVS = 34.1 and 32.3, P7GPG = 34.6 and 31.9, and GGPG = 31.3 and 28.1%). Median days in milk at first AI was affected by treatment (P7GPG = 59 vs. OVS = 68 and GGPG = 68 d). In experiment 2, GGPG reduced ED (GGPG = 23.3 vs. P7GPG = 74.9 and P11GPG = 79.6%). Treatment did not affect overall P/AI at 36 and 66 d after AI (GGPG = 29.2 and 25.8, P7GPG = 28.7 and 26.6, and P11GPG = 31.9 and 30.2%) or pregnancy loss. Cows artificially inseminated upon ED had greater P/AI than TAI cows (ED = 32.3 and TAI = 25.1%). However, treatment did not affect P/AI for cows artificially inseminated upon ED at 36 and 66 d after AI (GGPG = 29.6 and 27.3, P7GPG = 29.4 and 28.1, and P11GPG = 35.7 and 33.7%) or TAI (GGPG = 29.1 and 25.3, P7GPG = 25.0 and 22.1, and P11GPG = 16.9 and 16.9%). Median days between NPD and AI was affected by treatment (GGPG = 10 vs. P7GPG = 4 and P11GPG = 7 d). Prostaglandin-based programs increased ED and reduced interval to first AI and between AI. Gonadotropin-releasing hormone-based programs increased the proportion of TAI cows. Cows artificially inseminated upon ED had increased P/AI compared with TAI cows. [ABSTRACT FROM AUTHOR]

The objective was to evaluate the effects of GnRH or PGF(2α)-based synchronization and resynchronization programs on fertility in lactating dairy cows. For experiment 1, cows (n=1,521) were presynchronized with 2 injections of PGF given at 36 and 50 DIM and assigned to 1 of 3 protocols: Ovsynch [OVS; n=552; GnRH injection, PGF(2α) injection 7 d later, GnRH injection 56 h later, and timed artificial insemination (TAI) 16 h later] beginning at 14 d after presynchronization (PS), GnRH-GnRH-PGF(2α)-GnRH (GGPG; n=402) treatment with a GnRH injection given 7 d after PS and OVS 7 d later, or PGF(2α)-GnRH-PGF(2α)-GnRH (P7GPG; n=567) treatment with a PGF(2α) injection given 7 d after PS and OVS beginning 7 d later. Experiment 2 cows (n=2,327) were assigned to 1 of 3 resynchronization protocols 7 d before nonpregnancy diagnosis (NPD): GGPG cows (n=458) received a GnRH injection at enrollment and OVS at NPD; P7GPG cows (n=940) received a PGF(2α) injection at NPD and OVS 7 d later; and P11GPG cows (n=929) received a PGF(2α) injection 3 d after NPD and OVS 11 d later. In both experiments, cows were artificially inseminated upon estrus detection (ED). In experiment 1, 52.3% of cows were artificially inseminated upon ED, with GGPG having reduced ED (GGPG=46.8 vs. OVS=50.7 and P7GPG=57.7%). Treatments did not affect overall pregnancy per artificial insemination (P/AI) at 36 and 66 d after AI (OVS=34.1 and 32.3, P7GPG=34.6 and 31.9, and GGPG=31.3 and 28.1%, respectively) or pregnancy loss but cows artificially inseminated upon ED had higher P/AI than cows undergoing TAI (ED cows=37.9 vs. TAI cows=28.8%). Treatment did not affect P/AI for cows artificially inseminated upon ED or TAI at 36 and 66 d after AI (OVS=34.1 and 32.3, P7GPG=34.6 and 31.9, and GGPG=31.3 and 28.1%). Median days in milk at first AI was affected by treatment (P7GPG=59 vs. OVS=68 and GGPG=68 d). In experiment 2, GGPG reduced ED (GGPG=23.3 vs. P7GPG=74.9 and P11GPG=79.6%). Treatment did not affect overall P/AI at 36 and 66 d after AI (GGPG=29.2 and 25.8, P7GPG=28.7 and 26.6, and P11GPG=31.9 and 30.2%) or pregnancy loss. Cows artificially inseminated upon ED had greater P/AI than TAI cows (ED=32.3 and TAI=25.1%). However, treatment did not affect P/AI for cows artificially inseminated upon ED at 36 and 66 d after AI (GGPG=29.6 and 27.3, P7GPG=29.4 and 28.1, and P11GPG=35.7 and 33.7%) or TAI (GGPG=29.1 and 25.3, P7GPG=25.0 and 22.1, and P11GPG=16.9 and 16.9%). Median days between NPD and AI was affected by treatment (GGPG=10 vs. P7GPG=4 and P11GPG=7 d). Prostaglandin-based programs increased ED and reduced interval to first AI and between AI. Gonadotropin-releasing hormone-based programs increased the proportion of TAI cows. Cows artificially inseminated upon ED had increased P/AI compared with TAI cows., (Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Short-term fasting of mature ewes during diestrus results in increased serum concentrations of progesterone and a delayed pre-ovulatory surge release of LH. To determine if these changes in reproductive hormones influence subsequent follicular development, mature ewes observed in estrus were assigned randomly to control (n=10) or fasted (n=15) groups. Control ewes had ad libitum access to feed, whereas fasted ewes were not fed from day 7 through 11 of their estrous cycle. Daily blood samples were collected from control and fasted ewes throughout the fasting period. Fasting increased (P<0.001) serum concentrations of progesterone (4.4 ng/mL versus 2.7 ng/mL [+/-0.3]). On day 12, all ewes were treated with 10mg of PGF(2alpha) and fasted ewes were returned to ad libitum feed. Ovaries were collected from ewes (n=5 each group) at 0 and 72 h following PGF(2alpha) in control and 0, 72 and 96 h in fasted ewes. Ovaries were weighed and small (< or =2mm), medium (3-4mm), and large (> or =5mm) follicles were enumerated. Total numbers of follicles were less (P<0.001) in fasted than fed ewes (14.6 versus 30.2 [+/-2.2]) at 0 h, but did not differ (P=0.9) when numbers of follicles were compared at similar times before the anticipated LH surge (i.e., at 72 h versus 96 h in control and fasted ewes, respectively). Within follicular size class, numbers of small and medium follicles were decreased (P=0.04) at 0 h in fasted ewes. Numbers of large follicles did not differ (P=1.0) between groups. Although numbers of small and medium ovarian follicles in fasted ewes recovered by 96 h to values comparable to fed ewes at 72 h following PGF(2alpha), serum concentrations of estradiol 17beta (P=0.08) and FSH (P=0.06) tended to be decreased in fasted ewes before the anticipated surge release of LH. Pituitary content of LH and FSH also tended to be lower (P< or =0.09) at 96 h in fasted ewes than at 72 h in control ewes, but did not differ (P> or =0.4) at hour 0 following PGF(2alpha). Hypothalamic and stalk median eminence contents of GnRH were not influenced (P> or =0.2) by fasting at any time period. Fasting during the luteal phase perturbs gonadotropin secretion and may influence fertility by causing a delay in ovarian follicle development.

Objectives were to determine if neuropeptide Y (NPY) had direct effects GnRH induced secretion of LH from the anterior pituitary gland, and if endogenous steroids modulated the effect of NPY. To accomplish these objectives, 15 Hereford heifers were assigned to one of three ovarian status groups: follicular, luteal, or ovariectomized. One animal from each of the three ovarian status groups was slaughtered on each of 5 days and anterior pituitary gland harvested. Anterior pituitary gland cells within ovarian status were equally distributed and randomly assigned to one of three cell culture treatments: no NPY or GnRH (control), 10 nM GnRH, or 100 nM NPY+10 nM GnRH. Anterior pituitary cell cultures were incubated with or without NPY for 4 h and further incubated for an additional 2 h with or without GnRH and supernatant collected for quantification of LH. Treatment of anterior pituitary cell cultures with GnRH or GnRH+NPY did not affect LH release in cultures obtained from follicular (S.E.=5%; P=0.58) or ovariectomized (S.E.=7%; P=0.22) heifers. Both GnRH and GnRH+NPY increased LH release from anterior pituitary cell cultures from heifers in the luteal phase (S.E.=14%; P < or = 0.05) compared to control cultures. Cultures from luteal phase heifers treated with GnRH did not differ from those treated with GnRH+NPY (P=0.34). These data provide evidence to suggest that effects of NPY on LH release may occur primarily at the level of the hypothalamus.

Twelve Hereford x Angus heifers, 2.5 year, 492 +/- 17 kg (BCS = 5 +/- 0.5), were randomly assigned to one of three supplements, stratified by calving date and calf sex. Treatments consisted of a daily equivalent of: (1) low undegradable intake protein (UIP) (L: 908 g per cow per day; 36% CP, 108 g UIP), (2) middle UIP (M: 908 g per cow per day; 36% CP, 165 g UIP), and (3) high UIP (H: 908 g M + 243 g feather meal per cow per day; 46% CP, 335 g UIP). Cows were fed sudan grass hay (7.3% CP, as fed) daily at 2% BW (as fed). Supplement was individually fed twice weekly from Week 2 to Week 11 post-partum. Cow body weight (BW), backfat (BF) and rumpfat (RF) thicknesses decreased in all cows, (P < 0.05) yet did not differ among treatments (P > 0.10). There were no differences among treatments in calf BW (P > 0.10). Serum insulin, blood urea nitrogen (BUN), milk components and yield did not differ among treatments (P > 0.10). Area under the curve (AUC) for serum LH was greater (P = 0.07) in H versus L and M (809 versus 599 and 607 +/- 69 U, respectively). No differences were observed in FSH AUC or mean serum concentrations (P > 0.10). Uterine pH did not differ among treatments or between supplement versus non-supplement days (P > 0.10). Serum progesterone remained below I ng/ml for all cows indicating absence of return to estrus. Under the conditions of this study, BW, BF, RF, serum insulin, BUN, milk components, and yield, uterine pH and serum FSH were not affected by level of UIP. However, supplement containing high levels of UIP enhanced GnRH-induced LH release.

We conducted three experiments to test various protocols for synchronizing estrus, ovulation, or both before insemination of heifers. In experiment 1, 23 controls received two PGF2alpha injections; 23 heifers were treated like the controls plus a norgestomet implant for 8 d, with the second PGF2alpha injection 24 h before implant removal; and 23 heifers were treated like the previous group plus 100 microg of GnRH 54 h after the second PGF2alpha injection. Although norgestomet and GnRH altered some estrual characteristics, conception rates in experiment 1 (n = 69) and experiment 2 (278 heifers receiving the same treatments as those in experiment 1) generally were not different among treatments. Reproductive outcomes were not improved by adding norgestomet and GnRH to a standard PGF2alpha protocol. In experiment 3, control heifers received PGF2alpha and were inseminated after detected estrus or at 72 to 80 h after a second injection of PGF2alpha given 14 d after the first injection. Select Synch heifers, treated with GnRH either 6 or 7 d before PGF2alpha were inseminated after detected estrus, whereas Ovsynch heifers were treated like Select Synch heifers but also received a second GnRH injection approximately 36 h after PGF2alpha and were inseminated 18 h later. Estrus detection and pregnancy rates after Ovsynch were less than those of controls, whereas conception and pregnancy rates did not differ between control and Select Synch heifers. Therefore, the Select Synch protocol was equivalent to a standard PGF2alpha protocol, whereas Ovsynch was inferior to both of those protocols.

The subacute dose-response effects of swainsonine (SW) consumption on immunocompetence and serum constituents of sheep in a nutrient-restricted state were investigated. Sheep (23 wethers, 5 ewes) were assigned to 1 of 5 SW treatments (0, 0.2, 0.4, 0.8, or 1.6 mg swainsonine/ kg bw/d). Swainsonine was delivered by feeding locoweed (Oxytropis sericea) with grama grass and alfalfa hays for a 28-d treatment period followed by a 21-d recovery period without locoweed. Body weights were measured weekly and behavioral changes were monitored for clinical signs of SW toxicity. Venous blood was collected weekly for lymphoblastogenesis and serum constituent analyses. Clinical signs (sluggishness, decreased responsiveness) of swainsonine toxicity were observed from d 14 to 35 in the 0.8 and 1.6 mg treatments. Subacute oral exposure did not appear to affect lymphoblastogenic analyses. Acute and subacute alterations in various serum constituents did indicate subclinical effects of SW ingestion. Linear, quadratic and cubic dose-response relationships were detected for some serum constituents (e.g., alkaline phosphatase, aspartate aminotransferase). Subacute SW consumption at the levels investigated does not seem to affect the immunocompetence of nutrient restricted sheep. The lack of change in serum alkaline phosphatase at the 0.2 mg SW/kg bw/d dose indicates the potential for a no adverse effect level of SW consumption in nutrient restricted sheep. In combination with measurable SW in serum, rises in serum alkaline phosphatase and aspartate aminotransferase activities, and declines in serum Fe and cholesterol during subacute exposure to SW establish these markers as potential indicators of subclinical SW toxicosis.

The activins (dimers of betaA or betaB subunits, encoded by the genes Inhba and Inhbb, respectively) are TGF-beta superfamily members that have roles in reproduction and development. Whereas mice homozygous for the Inhba-null allele demonstrate disruption of whisker, palate and tooth development, leading to neonatal lethality, homozygous Inhbb-null mice are viable, fertile and have eye defects. To determine if these phenotypes were due to spatiotemporal expression differences of the ligands or disruption of specific ligand-receptor interactions, we replaced the region of Inhba encoding the mature protein with Inhbb, creating the allele Inhbatm2Zuk (hereafter designated InhbaBK). Although the craniofacial phenotypes of the Inhba-null mutation were rescued by the InhbaBK allele, somatic, testicular, genital and hair growth were grossly affected and influenced by the dosage and bioactivity of the allele. Thus, functional compensation within the TGF-beta superfamily can occur if the replacement gene is expressed appropriately. The novel phenotypes in these mice further illustrate the usefulness of insertion strategies for defining protein function.

The rate-limiting step in luteal biosynthesis of progesterone consists of cleavage of the side chain of cholesterol by mitochondrial cytochrome P450 side-chain cleavage enzyme (P450scc) to form pregnenolone. Luteal mRNA encoding P450scc, quantitated on selected days of the 16-day ovine estrous cycle, was similar on days 3 and 6, increased by 2-fold on day 9 (P < 0.05) and remained elevated on day 15. Levels of P450scc mRNA on day 15 of pregnancy were not different from those found on any day of the cycle (P < 0.05). To determine whether levels of mRNA encoding P450scc are hormonally regulated, ewes on day 10 of the estrous cycle were injected with hCG or prostaglandin F2 alpha (PGF2 alpha). P450scc mRNA was not increased for up to 36 h after injection of hCG, nor decreased within 8 h after injection of PGF2 alpha (P < 0.05). An assay for P450scc activity was developed which utilized ovine small and large luteal cells in the presence of 22R-hydroxycholesterol and ovine high density lipoprotein. Enzyme activity was quantitated by measurement of progesterone production. In small luteal cells activation of the protein kinase A (PKA) second-messenger system by treatment with LH resulted in 910% increase in progesterone production without altering activity of P450scc. Activation of the protein kinase C (PKC) second-messenger system with phorbol 12-myristate 13-acetate caused a 51% reduction in progesterone secretion from large luteal cells but did not alter activity of P450scc. These findings suggest that in mature luteal tissue steady state levels of mRNA encoding P450scc, and enzyme activity are independent of acute regulation by activation of PKA or PKC second-messenger systems.

Three experiments were conducted to determine how steady-state levels of mRNA encoding 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase (3 beta-HSD) in the ovine corpus luteum vary 1) between the two steroidogenic luteal cell types, 2) during the estrous cycle, and 3) during prostaglandin F2 alpha (PGF2 alpha)-induced luteolysis. In the first experiment, RNA (10 micrograms) was isolated from purified preparations (n = 4) of large or small steroidogenic luteal cells. Large luteal cells contained 42% more (p < 0.05) message for 3 beta-HSD per microgram RNA than did small luteal cells, while the amount of mRNA for tubulin did not differ between the two types of luteal cells. To determine whether luteal levels of mRNA for 3 beta-HSD differ during the estrous cycle, corpora lutea were collected from cycling ewes (n = 3/day) on Days 3, 6, 9, 12, and 15 postestrus. Levels of mRNA for 3 beta-HSD were similar on Days 3, 6, 9, and 12 but were lower (p < 0.05) on Day 15 postestrus, while levels of mRNA for tubulin were unchanged. In the final experiment, ewes were treated on Day 10 postestrus with two injections of PGF2 alpha (5 mg each) or saline (control) at a 4-h interval. Corpora lutea were collected from ewes (n = 4/treatment) 1 h or 8 h after the second injection of PGF2 alpha or 8 h after the second saline injection.(ABSTRACT TRUNCATED AT 250 WORDS)