The variation in the beta-casein genotypes and its effect on milk yield and genomic values in Holstein-Friesian cows.

The A2 milk marker is gaining popularity worldwide; thus, many farms plan to convert their dairy cattle herds to the A2A2 genotype. Variation in beta-casein genotypes needs to be monitored in large dairy cattle populations. Therefore we aimed to evaluate the genotypic distributions, population genetics, and diversity parameters in Holstein-Friesian cows. A total of 1200 cattle were genotyped using the Affymetrix® Axiom® array system. We performed an association analysis regarding the CSN2 genotypes and phenotypic traits, including lactation and test-day milk yield. We next evaluated the effects of the genotypes considering the genetic merit of the animals. Animals were grouped based on their PTAs for milk production, fat, protein, and daughter pregnancy rate. Thus, we tested the genotype × genetic merit interaction for significance. The A2 allele frequency is remarkably high (0.68), and the heterozygous genotype is predominant (46.25%). The marker showed intermediate variability and diversity levels, indicating a considerable frequency of the A1A1 genotype (9.33%) remains in the population. ANOVA results showed no significant association between the CSN2 genotypes and milk yield traits. A similar finding is valid for the genotype × genetic merit regarding the genomic test results. The data presented here may be helpful for further investigations and applications on A2 milk production.

Effects of daily propylene glycol drenching during the Ovsynch protocol on fertility and metabolic parameters in lactating dairy cows.

Negative energy balance (NEB) caused by restricted feed intake leads to body condition loss (BCS), increased metabolic stress and reduced fertility in dairy cows. Propylene glycol (PG) is a precursor of ruminal propionate for gluconeogenesis used to increase metabolic adaptation to the early postpartum period. The aim of this study was to determine the effects of daily drenching of PG during the fixed-time artificial insemination (FTAI) protocol on beta-hydroxybutyric acid (BHBA), glucose, adiponectin, insulin-like growth factor-1 (IGF1) concentrations, follicle size and pregnancy rate in dairy cows. Cows (n = 148) were randomly divided into two groups and received either 300 mL of PG (PG-OVS, n = 76) or 300 mL of water (CON-OVS, n = 72) each day of the Ovsynch protocol (GnRH-7 days-PGF2α -56 hours-GnRH-16-18-hour FTAI) between days 57 ± 3 to 67 ± 3 postpartum for the first service. Body condition scores (14 days before expected calving, at calving, on days 21 and 42 postpartum) were recorded. Blood samples were collected days 7 ± 3, 21 ± 3 postpartum, at the beginning of the Ovsynch (days 57 ± 3) and at the time of FTAI (days 67 ± 3) for measurements of BHBA, glucose, adiponectin and IGF1 concentrations. Ultrasonographic examinations were done to measure follicle size at the beginning of Ovsynch and FTAI and to determine pregnancy on days 30 and 60 following FTAI. There were no differences (p > .05) in glucose, adiponectin and IGF1 concentrations between the groups during the study. Although there was no difference (p > .05) in BHBA concentrations on postpartum day 7 ± 3, 21 ± 3 and 57 ± 3 between the groups, BHBA concentrations at the time of insemination was lower (p < .05) in the PG-OVS group (0.72 ± 0.03 mmol/L) than in the CON-OVS group (0.81 ± 0.03 mmol/L). Follicle sizes at the beginning of Ovsynch (PG-OVS, 14.5 ± 0.48 mm; CON-OVS, 14.3 ± 0.59 mm) and FTAI (PG-OVS, 17.8 ± 0.52 mm; CON-OVS, 17.7 ± 0.42 mm) were not different (p < .05). Pregnancy rate of the cows in the PG-OVS group (46.1%, 35/76) was higher (p = .05) than in the CON-OVS group (30.6%, 22/72) on day 30 following FTAI. In conclusion, decreasing serum BHBA concentrations at the time of FTAI by means of daily drenching of PG during the Ovsynch protocol, increased the pregnancy rate at first service in lactating dairy cows. On the other hand, blood glucose was not related with pregnancy rates in our study, probably as a result of our sampling time and more rapid fluctuations of blood glucose concentrations when compared to BHBA.

Effect of oestrous expression prior to timed artificial insemination with sexed semen on pregnancy rate in dairy cows.

The objectives of the study were to determine (1) oestrous expression rate and (2) the effect of oestrous expression prior to progesterone-based Ovsynch protocol on pregnancy rate in Holstein cows. All cows (n = 917) were subjected to 7-day progesterone-based Ovsynch protocol. In this protocol, cows that expressed oestrus before (HEAT1) the scheduled second GnRH were inseminated 20 h later after the onset of oestrus without GnRH administration. Cows that expressed oestrus after the second GnRH administration (HEAT2) or did not express oestrus (NOHEAT) received fixed-timed AI. Oestrous expression was determined by using activity-rumination monitoring system and all cows were inseminated with sexed semen. Oestrous expression rate prior to FTAI was 40.5% and the majority (p < .01) of oestrous expression were in HEAT2 compared with HEAT1 in both primiparous (71.8 vs. 28.1%) and multiparous cows (69.5 vs. 30.5%). The mean interval from intravaginal device removal to the onset of oestrus was 47.4 ± 0.9 h and 62.9 ± 0.5 in HEAT1 and HEAT2, respectively. Primiparous cows (47.7%) had a higher (p < .01) expression rate compared with multiparous cows (37.2%). Overall pregnancy rate was 37.4% and there was two-way significant interaction between parity and oestrous expression on pregnancy rate (p < .01). Both primiparous (48.1 vs. 35.8%) and multiparous cows (47.4 vs. 28.4%) that expressed oestrus had greater (p < .01) pregnancy rate compared with cows that did not express oestrus. There was no difference in pregnancy rates of HEAT1 and HEAT2 in both primiparous (44.7 vs. 49.5%) and multiparous cows (47.2 vs. 47.6%). Pregnancy rate was not influenced (p = .21) by milk production (high or low) in both primiparous (47.6 vs. 48.6%) and multiparous (54.9 vs. 42.1%) cows that expressed oestrus, respectively. In conclusion, cows showing oestrus before or after second GnRH of the Ovsynch protocol had greater pregnancy rate than cows not showing oestrus.

Comparison of pregnancy per AI of heifers inseminated with sex-sorted or conventional semen after oestrus detection or timed artificial insemination.

The objective of the study was to compare the fertility after using sex-sorted or conventional semen either with oestrus detection (EST) or timed artificial insemination (TAI) in Holstein heifers. Holstein heifers were randomly assigned to one of the following treatments in a 2 × 2 factorial design. Heifers in the EST group were inseminated with sex-sorted (n = 114) or conventional semen (n = 100) after spontaneous or induced oestrus. Heifers in the TAI, subjected to the 5-day Cosynch+Progesterone protocol (GnRH+P4 insertion-5d-PGF2α +P4 removal-1d-PGF2α -2d-GnRH+TAI), were inseminated with sex-sorted (n = 113) or conventional semen (n = 88). Statistical analyses were performed using PROC GLIMMIX procedure of SAS 9.4 (SAS Institute Inc., Cary, NC). Overall P/AI was 60.7% for EST and 54.2% for TAI regardless of types of semen and 68.1% for conventional and 48.9% for sex-sorted semen regardless of insemination strategies. Fertility of heifers inseminated with either sex-sorted (53.5%; 44.2%) or conventional (69.0%; 67.0%) semen did not differ between EST and TAI respectively. Besides, the interaction between the semen type and the insemination strategy was not significant for P/AI. The embryonic loss was significantly greater with sex-sorted semen (17.1%) compared to conventional semen (1.6%). There was no sire effect with sex-sorted semen on P/AI (52.6% vs. 46.2%) and embryonic loss (16.4% vs. 18.0%). As expected, sex-sorted semen resulted in more female calves (89.8% vs. 51.6%) than conventional semen. Thus, sex-sorted semen can be used with 5-day Cosynch+Progesterone protocol to eliminate the inadequate oestrus detection and to increase female calves born in dairy heifers.

Effect of delaying the time of insemination with sex-sorted semen on pregnancy rate in Holstein heifers.

The objective of the study was to evaluate the interval from onset of oestrus to time of artificial insemination (AI) to obtain the optimum pregnancy rate with sex-sorted semen in Holstein heifers. Heifers in oestrus were detected and inseminated only by using heat-rumination neck collar comprised electronic identification tag at the age of 13-14 months. Heifers (n = 283) were randomly assigned to one of three groups according to the timing of insemination at 12-16 hr (G1, n = 97), at 16.1-20 hr (G2, n = 94) and at 20.1-24 hr (G3, n = 92) after reaching the activity threshold. The mean duration of oestrus was 18.6 ± 0.1 hr, and mean peak activity was found at 7.5 ± 0.1 hr after activity threshold. The mean interval from activity threshold to ovulation was 29.4 ± 0.4 hr. The overall pregnancy per AI (P/AI) was 53.0% at 29-35 days and 50.9% at 60-66 days after AI. There was a significant reduction between G1 (13.8 ± 1.4 hr) and G3 (7.9 ± 1.4 hr) related to the intervals from AI to ovulation time. Sex-sorted semen resulted in significantly higher P/AI at 29-35 days when heifers inseminated in G3 (60.9%) after oestrus than those inseminated in G1 (49.5%) and G2 (48.9%). In terms of fertility, when the temperature-humidity index (THI) was below the threshold value (THI ≤65) at the time of AI, there was a tendency (≤65; 57.2% vs. > 65; 47.1%) for high pregnancy rate. There was no effect of sire on P/AI. In addition, the interaction of the technician with the time of AI was found significant, and three-way interaction of technician, sire and time of AI was tended to be significant on pregnancy rate. Thus, in addition to delaying the time of insemination (between 20.1 and 24 hr) after oestrous detection, THI and experienced technician were also found to be critical factors in increasing fertility with the use of sex-sorted semen in Holstein heifers.

Effect of presynchronization with prostaglandin F2α before the 5-d timed AI protocol on ovarian responses and pregnancy in dairy heifers.

The objectives were the determine the effects of presynchronization with PGF2α 2 days before the 5-d timed artificial insemination (AI) protocol on ovarian responses and pregnancy per AI (P/AI) in dairy heifers. The hypothesis was that PGF2α would induce responsive heifers to be in proestrus at the initiation of the timed AI protocol, which was expected to improve ovulatory responses and P/AI. Weekly cohorts of Holstein heifers were blocked by age and, within block, randomly assigned to remain as control (CON; n = 255) or receive PGF2α on experiment Day -10 (PG; n = 255). All heifers were subjected to the 5-d timed AI protocol (Day -8, GnRH + intravaginal progesterone controlled internal drug release insert; Day -3, PGF2α and insert removal; Day -2, PGF2α; and Day 0, GnRH and AI). A subset of 22 blocks of heifers (n = 43) had their ovaries scanned by ultrasonography on experiment Days -8, -3, 0, and 2 and blood was sampled and analyzed for concentrations of progesterone on experiment Days -8, -7, -5, and -3. Pregnancy was diagnosed on experiment Days 32 and 60. On the day of the first GnRH of the timed AI protocol, PG heifers had smaller concentration of progesterone in plasma (CON = 4.5 ±â€¯0.5 vs. PG = 0.5 ±â€¯0.5 ng/mL), but larger follicular diameter (CON = 9.1 ±â€¯0.5 vs. PG = 11.0 ±â€¯0.5 mm), and a greater proportion of them had a follicle with at least 8.0 mm in diameter (CON = 61.9 vs. PG = 90.9%) than CON heifers, which resulted in increased ovulation to GnRH (CON = 19.0 vs. PG = 86.3%). Ovulation to the initial GnRH of the protocol increased as the concentration of progesterone in plasma decreased, from less than 20% when progesterone was greater than 5.0 ng/mL to more than 65% when progesterone was less than 1.0 ng/mL. More CON than PG heifers spontaenously ovulated before the day of timed AI. Detection of estrus on the day of timed AI did not differ between treatments (CON = 50.9 vs. PG = 46.6%), but P/AI on Days 32 (CON = 52.9 vs. PG = 61.1%) and 60 (CON = 49.0 vs. PG = 57.1%) after insemination tended to be greater for PG than CON; however, the benefit to presynchronization was observed in heifers inseminated with conventional (CON = 54.7 vs. PG = 67.4%), but not in heifers inseminated with sex-sorted semen (CON = 50.9 vs. PG = 52.8%). Administration of PGF2α 2 days before initiating the timed AI protocol induced heifers to be in proestrus, which enhanced ovulation to the initial GnRH and favored pregnancy per AI, particularly in heifers inseminated with conventional semen.

Fertility of lactating dairy cows inseminated with sex-sorted or conventional semen after Ovsynch, Presynch-Ovsynch and Double-Ovsynch protocols.

The objective was to compare pregnancy per artificial insemination (P/AI) with conventional (CS) or sex-sorted semen (SS) in dairy cows subjected to one of the three timed AI protocols. Cows (n = 356) were randomly assigned to synchronization with Ovsynch (OVS), Presynch-Ovsynch (PO) or Double-Ovsynch (DO) and inseminated on Day 77 ± 3 postpartum with either frozen-thawed SS (n = 182) or CS (n = 184) of the same bull. More cows were cyclic at the beginning of breeding Ovsynch increased (p < 0.01) with presynchronization and it was greater for DO than PO (OVS = 78.5%, PO = 85.1%, DO = 95.6%). Overall, P/AI for SS and CS increased with presynchronization (p < 0.05) on Days 31 (OVS = 35.5%, PO = 47.1%, DO = 48.3%) and 62 (OVS = 30.1%, PO = 43.8%, DO = 43.9%). Regardless of synchronization treatments, insemination with SS reduced P/AI (p < 0.02) on Days 31 (38.1% vs. 50.6%) and 62 (34.5% vs. 45.6%) compared with CS. No interaction was observed between synchronization treatment and type of semen for P/AI, although in cows receiving CS, P/AI was numerically greatest for PO (OVS = 42.0%, PO = 59.3%, DO = 49.0%), and in cows receiving SS, it was numerically greatest for those inseminated following DO (OVS = 27.9%, PO = 35.5%, DO = 47.6%). Thus, presynchronization improved P/AI in cows inseminated with sex-sorted or conventional semen.

Comparison of synchronisation and fertility after different modifications of the ovsynch protocol in cyclic dairy cows.

The objectives of this study were to evaluate the effects of (1) double doses of PGF2α administration or (2) an exogenous progesterone (CIDR) applied concurrently with, or (3) the day after, first GnRH of Ovsynch (GnRH-1), on synchronisation and fertility during the Ovsynch protocol. All cows (n = 378) received the Ovsynch protocol (GnRH-7d-PGF2α-56h-GnRH-18h-TAI). The 'OVS' group (n = 105) received only the Ovsynch protocol. The 'OVS-PGF' group (n = 118, GnRH-7d-PGF2α-12h-PGF2α-44h-GnRH-18h-TAI) received an extra dose of PGF2α 12 h later on Day 7. The 'OVS-7CIDR' group (n = 78, GnRH+CIDR-7d-PGF2α-56h-GnRH-18h-TAI) received a CIDR for 7 days between GnRH-1 and PGF2α. In the 'OVS-6CIDR' group (n = 77, GnRH-24h-CIDR-6d-PGF2α-56h-GnRH-18h-TAI), CIDR was applied one day after GnRH-1 and removed 6 days later. When all cows were evaluated, the responses to GnRH-2 were higher (P = 0.005) in cows that responded to GnRH-1 (95.4%) compared to the cows that did not respond (87.6%). The pregnancy rates at 31 and 62 days for each group were 48.6% and 42.9% in the OVS, 54.2% and 52.5% in the OVS-PGF, 52.6% and 48.7% in the OVS-7CIDR, and 55.8% and 49.3% in the OVS-6CIDR groups. Thus, none of the three different treatments has an effect on increasing the out-comes of the Ovsynch protocol in cyclic lactating dairy cows.

The role of luteinizing hormone in regulating gene expression during selection of a dominant follicle in cattle.

At approximately 8.5 mm in diameter, the future dominant follicle is "selected" for continued growth in cattle. In the present study, cows were treated with a gonadotropin-releasing hormone receptor antagonist, acyline, just before follicle selection (near 7.8 mm) to investigate the role of LH in changing mRNA concentrations during selection of a dominant follicle. The ovaries containing the expected dominant follicle (EDF; first largest follicle) and expected largest subordinate follicle (ESF) were removed after 12 or 24 h of treatment. Real-time PCR was used to determine mRNA concentrations. ELISA was used to measure testosterone and 17beta-estradiol (E(2)) and radioimmunoassay to measure androstenedione (A(4)) in follicular fluid. Concentrations of E(2) were greater in EDF than in ESF of untreated cows near the time of follicle selection (12 h) or at 12 h after selection (24 h). Testosterone, E(2), and A(4) were all dramatically decreased by acyline treatment at both times. In theca cells, acyline treatment reduced CYP17A1 (P450 17alpha) in EDF and STAR (steroidogenic acute regulatory protein) in both EDF and ESF but did not alter CYP11A1 (P450scc). In granulosa cells (GCs), LHCGR (luteinizing hormone [LH] receptor) was much greater in EDF than in ESF at both time of selection (739% greater) and 12 h after selection (2837% greater) and was decreased by acyline in EDF (87% decrease). The mRNA for CYP19A1 (cytochrome P450 aromatase) and PAPPA (pregnancy-associated plasma protein-A) tended to be greater in EDF than in ESF at follicle selection, and both mRNAs were much greater at 12 h after selection, with acyline significantly decreasing PAPPA mRNA after 24 h of treatment. The mRNA for FSHR (follicle-stimulating hormone receptor) was not different in EDF versus ESF and was not altered by acyline. Thus, induction of LHCGR mRNA in GCs is an early event during the follicle selection process, and surprisingly, expression of LHCGR mRNA is dependent on circulating LH. Production of follicular A(4), testosterone, and E(2) are also acutely related to LH but due to changes in expression of STAR and CYP17A1 in TC.

Pathological abnormalities after long-term administration of medroxyprogesterone acetate in a queen.

In this report, multiple abnormalities (bilateral ovarian cysts, cystic endometrial hyperplasia and pyometra (CEH-P), mammary adenoma, fibrosarcoma and cystic-papillary adenocarcinoma) identified in a queen continually administered medroxyprogesterone acetate (MPA) for 9 years are described. An 11-year-old domestic shorthair intact queen was presented for reduced appetite, polyuria, polydipsia, abdominal distension, and mammary mass. Pyometra was diagnosed based upon clinical, ultrasonographic and radiographic examinations. Mastectomy and ovariohysterectomy (OHE) were performed for treatment. Bilateral ovarian cysts were detected during OHE. Histopathologically, follicular ovarian cysts, CEH-P, and benign and neoplastic mammary lesions were identified. We suggest that, the pathological abnormalities may have been attributed to adverse effects of prolonged administration of MPA.

Changes in reproductive physiology of lactating dairy cows due to elevated steroid metabolism.

This manuscript focuses on potential changes in reproductive physiology that occur due to high milk production in lactating dairy cows. Four reproductive measures are discussed: interval to first ovulation, conception rate, duration of estrus, and multiple ovulation rate. The last two responses have now been closely linked to level of milk production. In contrast, time to first ovulation does not appear to be associated with level of milk production, and the association of conception rate with level of milk production is still controversial. In an attempt to explain some of the changes in reproductive physiology caused by high milk production a model of elevated steroid metabolism in lactating dairy cows is presented. Although many aspects of this model remain to be tested, a central role for elevated steroid metabolism in lactation-induced reproductive changes seems likely.

Follicular cysts occur after a normal estradiol-induced GnRH/LH surge if the corpus hemorrhagicum is removed.

The pathophysiology underlying follicular cysts appears to be lack of an estradiol (E2)-induced GnRH/LH surge due to hypothalamic insensitivity to E2. In addition, progesterone (P4) can cause animals with follicular cysts to resume normal cyclicity and normal hypothalamic responsiveness to E2. We postulated that follicular cysts may be a pathological manifestation of a physiological state that cows, and possibly other species, go through during the normal estrous cycle but the rise in P4 following ovulation induces them back to normal hypothalamic responsiveness to E2. Based on this hypothesis, we expected that removal of the ovary containing the corpus hemorrhagicum would prevent the normal rise in P4 following ovulation and induce development of follicular cysts. Cows (n = 24) on day 7 of the estrous cycle were treated with prostaglandin F2alpha (PGF2alpha) and time of ovulation was detected by ovarian ultrasonography every 8 h. Immediately following detection of ovulation, cows were randomly but unequally assigned to have the ovary containing the corpus hemorrhagicum removed (TRT; n = 16) or the ovary opposite to the corpus hemorrhagicum removed (CON; n = 8). Cows were subsequently evaluated by daily ultrasound and blood sampling to determine follicular dynamics. Ovulation was detected at 93.7 +/- 4.5 h after PGF2alpha injection. All CON cows had a normal estrous cycle length (22.0 +/- 0.6 days) after ovariectomy (OVX). Half of the TRT cows became anovular (TRT-ANO; n = 8) after OVX with large anovular follicles developing on the ovary (maximal size, 25.4 +/- 1.4 mm; range, 20-32 mm). However, eight TRT cows ovulated (TRT-OV; n = 8) 7.3 +/- 0.6 days after OVX. Control cows had a normal P4 rise after ovulation. Removal of the newly formed corpus hemorrhagicum prevented the rise in circulating serum P4 in TRT-ANO cows throughout the 25-day experimental period. The TRT-OV cows had a delayed increase in circulating P4 but it was normal in relation to time of ovulation. Serum E2 concentrations were similar among groups (TRT-OV, TRT-ANO and CON cows) until 7 days after OVX. Serum E2 concentrations then decreased in TRT-OV and CON cows but remained elevated (>5 pg/ml) in TRT-ANO cows. Thus, the endogenous increase in circulating E2 that induces the GnRH/LH surge and estrus is sufficient to induce cows into a physiological state that resembles follicular cysts if it is not followed by increased circulating P4.

Length of progesterone exposure needed to resolve large follicle anovular condition in dairy cows.

Hypothalamic unresponsiveness to an estradiol surge appears to be an underlying cause of large follicle anovular condition (follicular cysts), but progesterone exposure for 7 days resolves this condition. In this study, dairy cows with induced (Experiment 1) or naturally occurring (Experiment 2) follicular cysts were treated for different times with progesterone. In Experiment 1, 16 of 26 cows (62%) were induced into anovulation by causing a GnRH/LH surge when no ovulatory follicle was on the ovary. Anovular cows (n = 16) were assigned to one of four treatment groups ( 0, 1, 3, or 7 days of progesterone treatment) using an intravaginal, progesterone-releasing implant (CIDR). All anovular cows had low circulating progesterone concentrations before controlled internal drug releasing (CIDR) and greater concentrations that reached steady state (1.3 +/- 0.1 ng/mL progesterone) by 3 h after CIDR insertion. Circulating progesterone decreased to basal concentrations by 4 h after CIDR removal. Cows were treated with 5mg estradiol benzoate (EB) 12 h after CIDR removal. None (n = 4) of the control cows (0 day) had an LH surge after EB. All of the 3 days (5/5) and 7 days (4/4) CIDR-treated cows had an LH surge following EB, but only one of the 1 day (1/3) CIDR-treated cows. Magnitude of the LH peak was similar in the 3 and 7 days cows. All cows treated for 7 days ovulated (4/4), whereas, ovulation occurred in only 3/5, 1/3, and 0/4 of the cows treated for 3, 1, and 0 day, respectively. The two cows in the 3 days group that did not ovulate had a normal LH surge, but these two cows had a smaller maximal follicle size than cows that ovulated. In Experiment 2, naturally anovular lactating dairy cows (24 of 248) were identified using weekly ultrasonography. All anovular cows grew follicles to >12 mm, with 54% (13 of 24) having follicles larger than ovular size (15-24 mm) and 33% (8 of 24) having follicles that would be considered cystic (>25 mm). Anovular cows were randomly assigned to CIDR treatment for 0, 1, or 3 days. All (7/7) of 3 days, 33% (3/9) of 1 day, and 25% (2/8) of control (0 day) cows ovulated by 1 week after CIDR removal. Thus, 3 days but not 1 day of progesterone exposure appears to be sufficient to reinitiate estradiol responsiveness of the hypothalamus.

Ovulation rate after GnRH or PGF2alpha administration in early postpartum dairy cows.

The objectives of this study evaluating induction of ovulation in early postpartum dairy cows were to: compare two methods of GnRH (100 mcg) administration (i.m. route and s.c. implant), and determine if prostaglandin F(2alpha) (PGF) causes release of LH or ovulation similar to that reported for GnRH. In trial #1, serum LH peaked at 2h after i.m. administration of GnRH and was declining at 4h. The s.c. GnRH implant also caused an elevation in serum LH at 2 and 4h after treatment, with LH declining at 6h. Serum LH was unchanged in control cows. Experimental treatment caused ovulation in 4 of 14 GnRH i.m. treated cows, 4 of 12 GnRH implanted cows and 0 of 13 control cows. Parity had no effect on LH response but did affect resulting ovulation rate as multiparous cows were more likely to ovulate than were primiparous cows in response to either GnRH treatment. All cows that ovulated had a follicle larger than 12 mm at the time of treatment. In trial #2, serum LH increased as before after i.m. administration of GnRH, however, serum LH was unchanged in cows treated with PGF or saline. Gonadotropin releasing hormone caused more cows to ovulate than did PGF or saline treatments, and GnRH shortened the interval from treatment to the onset of CL function over the PGF treatment; 13.9+/-2.6, 28.2+/-4.1 and 22.3+/-4.1 days for GnRH, PGF and saline, respectively. In summary, there was no difference in the ability of s.c. implantation and i.m. administration of GnRH to cause ovulation. Prostaglandin F(2alpha) did not cause release of LH or ovulation. In 22 early postpartum dairy cows treated with 100 mcg GnRH i.m. in these two trials, nearly all cows (95%) responded with a release of LH but only 45% (10/22) responded with an ovulation and subsequent formation of a CL.

An alteration in the hypothalamic action of estradiol due to lack of progesterone exposure can cause follicular cysts in cattle.

Many mammals, including cattle, can develop ovarian follicular cysts, but the physiological mechanisms leading to this condition remain undefined. We hypothesized that follicular cysts can develop because estradiol will induce a GnRH/LH surge on one occasion but progesterone exposure is required before another GnRH/LH surge can be induced by estradiol. In experiment 1, 14 cows were synchronized with an intravaginal progesterone insert (IPI) for 7 days, and prostaglandin F(2alpha) was given on the day of IPI removal. Estradiol benzoate (EB; 5 mg i.m.) was given 3 days before IPI removal to induce atresia of follicles. Cows were given a second EB treatment 1 day after IPI removal to induce a GnRH/LH surge in the absence of an ovulatory follicle. All cows had an LH surge following the second EB treatment, and 10 of 14 cows developed a large-follicle anovulatory condition (LFAC) that resembled follicular cysts. These LFAC cows were given a third EB treatment 15 days later, and none of the cows had an LH surge or ovulation. Cows were then either not treated (control, n = 5) or treated for 7 days with an IPI (n = 5) starting 7 days after the third EB injection. Cows were treated for a fourth time with 5 mg of EB 12 h after IPI removal. All IPI-treated, but no control, cows had an LH surge and ovulated in response to the estradiol challenge. In experiment 2, cows were induced to LFAC as in experiment 1 and were then randomly assigned to one of four treatments 1) IPI + EB, 2) IPI + GnRH (100 microg), 3) control + EB, and 4) control + GnRH. Control and IPI-treated cows had a similar LH surge and ovulation when treated with GnRH. In contrast, only IPI-treated cows had an LH surge following EB treatment. Thus, an initial GnRH/LH surge can be induced with high estradiol, but estradiol induction of a subsequent GnRH/LH surge requires exposure to progesterone. This effect is mediated by the hypothalamus, as evidenced by similar LH release in response to exogenous GnRH. This may represent the physiological condition that underlies ovarian follicular cysts.