Pregnancy Rates in Angus Cross Beef Cows Bred at Observed Oestrus With or Without Second GnRH Administration in Fixed‐Time Progesterone‐Supplemented Ovsynch and CO‐Synch Protocols

Crossbred cows (n = 1073) from five locations had oestrous cycles synchronized with 100 μg of GnRH IM and insertion of controlled internal drug release device (CIDR) on Day 0 followed by 25 mg of PGF_2α IM and CIDR removal on Day 7. Kamar^® patches were placed on all cows at CIDR removal. Cows were observed three times daily for oestrus after PGF_2α administration. In the Ovsynch‐CIDR group, cows detected in oestrus (n = 193) within 48 h after PGF_2α were inseminated using the AM–PM rule. Among these cows, 80 received and 113 did not receive a second GnRH at 48 h after PGF_2α. Cows (n = 345) not detected in oestrus received a second GnRH at 48 h after PGF_2α on Day 9, and fixed‐time AI 16 h after the GnRH on Day 10. In the CO‐Synch‐CIDR group, cows detected in oestrus (n = 224) within 48 h after PGF_2α were inseminated using the AM–PM rule. Among these cows, 79 received and 145 did not receive a second GnRH at 64 h after PGF_2α. Cows (n = 311) not detected in oestrus received a second GnRH on Day 10 at the time of AI, 64 h after PGF_2α. The AI pregnancy rates were not different between the Ovsynch‐CIDR and CO‐Synch‐CIDR groups (p = 0.48). There were no differences in the AI pregnancy rates for cows inseminated at a fixed time (p = 0.26) or at detected oestrus (p = 0.79) between the treatment groups. Among cows inseminated in oestrus, there were no differences in the AI pregnancy rates between cows that received or did not receive the second GnRH (p = 0.47). In conclusion, acceptable AI pregnancy rates can be achieved with or without inclusion of oestrus detection in the Ovsynch‐CIDR and CO‐Synch‐CIDR protocols. Among cows detected in oestrus, cows that received a second GnRH yielded similar pregnancy rates when compared with cows that did not receive the second GnRH.     

Effect of Acute Bull Exposure Around the Time of Artificial Insemination on Serum Oxytocin and Progesterone Concentrations and Pregnancy Rates in Dairy Cows

This study examined the effect of acute bull exposure around the time of artificial insemination (AI) on oxytocin and progesterone concentrations, and pregnancy rates in dairy cows. Ninety six dairy cows, stratified according to parity into primiparous and pluriparous, were divided into three groups; short bull exposure (SBE; 10 min, n = 32), long bull exposure (LBE; 4 h, n = 32) or no bull exposure (NBE; n = 32). On day 45 post‐partum, all cows were treated with PGF2α on three occasions 11–14 days apart to synchronize oestrus. They were submitted to fixed time AI 80 h after the third PGF2α injection. Cows in the SBE and LBE groups were artificially inseminated 5 min after the introduction of the bull. From a subset of cows (n = 6 per group; three primiparous and three pluriparous), blood samples were collected once every 5 min starting 15 min before AI until 15 min after AI and analysed for oxytocin concentrations. Additional blood samples were collected for measurements of progesterone (P4) concentrations once daily for 4 days starting on the day of AI and once every 3 days thereafter until day 22. The effects of bull exposure, time, parity, difficulty of AI, and pregnancy on oxytocin and P4 concentrations were analysed using the mixed linear model procedure. Mean oxytocin concentrations or change in oxytocin concentrations after bull exposure or AI were not different among groups. Pregnancy rates for the NBE, SBE and LBE groups were 55.5%, 33.3% and 44.4%, respectively, and were not different among groups. In conclusion, acute bull exposure around the time of AI did not affect oxytocin and progesterone concentrations and did not improve pregnancy rates in dairy cattle under these farms conditions.     

Is a Delayed Treatment with GnRH,hCG or Progesterone Beneficial for Reducing Embryonic Mortality in Buffaloes?

The aims of this study were to verify the efficacy of delayed hormonal treatments performed on day 25 post‐insemination on pregnancy rate at 45 and 70 days in buffalo. The trial was performed on 385 buffaloes synchronized by the Ovsynch/TAI protocol and submitted to artificial insemination (AI). Twenty‐five days after AI, pregnant animals were assigned to four treatments: (1) GnRH agonist (n = 52), 12 μg of buserelin acetate; (2) hCG (n = 51), 1500 IU of human chorionic gonadotrophin; (3) Progesterone (n = 47), 341 mg of P4 intramuscular (im) every 4 days for three times; (4) Control (n = 54), treatment with physiological saline (0.9% NaCl). Milk samples were collected on days 10, 20 and 25 after AI in all buffaloes to determine progesterone concentration in whey by radioimmunoassay method. Statistical analysis was performed by anova . Pregnancy rate on day 25 after AI was 52.9%, but declined to 41.8% by day 45, indicating an embryonic mortality (EM) of 21%. If only control group is considered, the incidence of EM was 38.9%. Pregnant buffaloes had higher (p < 0.01) progesterone concentrations on day 20 and 25 after AI than both non‐pregnant buffaloes and buffaloes that showed EM. The treatments on day 25 increased (p < 0.01) pregnancy rate, although in buffaloes with a low whey progesterone concentration on day 20 and 25 after AI (n = 22); all treatments were ineffective to reduce EM.     

Use of Vaginal Electrical Resistance to Diagnose Oestrus,Dioestrus and Early Pregnancy in Synchronized Tropically Adapted Beef Heifers

The primary objective of this study was to determine whether a single measurement of intravaginal electrical resistance (VER), using the commercially available Ovatec^® probe, can discriminate between dioestrus and oestrus in Bos indicus females, which had been treated to synchronize oestrus. Santa Gertrudis heifers (n = 226) received one of three oestrous synchronization treatments: double PGF_2α 10 days apart, 8‐day controlled internal drug release (CIDR) treatment or CIDR pre‐synchronization + PGF_2α 10 days after CIDR removal. The heifers were inseminated within 12 h following observed oestrus, or, if not observed, at a fixed time approximately 80 h, following the last synchronization treatment. They were palpated per rectum for signs of pregnancy 9 weeks after artificial insemination (AI). Vaginal electrical resistance measurements were taken at the completion of synchronization treatments (presumed dioestrus), immediately prior to AI (oestrus), and then at 3 and 9 weeks post‐AI. Mean VER differed between presumed dioestrus and oestrus (113.7 vs 87.4, p < 0.001). The area under the receiver operating characteristics (ROC) curve was 0.925, indicating that VER was highly discriminatory between dioestrus and oestrus. Vaginal electrical resistance at time of AI was negatively associated with odds of conception when all inseminations were included in the analyses [odds ratio (OR) = 0.97; 95% CI 0.95–1.00; p = 0.018], but not when fixed time AIs were excluded (OR = 1.00; 95% CI 0.97–1.03; p = 0.982). Mean VER readings differed between pregnant and non‐pregnant animals at both 3 weeks (120.5 vs 96.7, p < 0.001) and 9 weeks (124.0 vs 100.3, p < 0.001) post‐AI. However, 3‐ and 9‐week VER measurements were not highly discriminatory between pregnancy and non‐pregnancy (area under ROC curve = 0.791 and 0.736, respectively). Mean VER at time of AI for animals diagnosed in oestrus differed between each of the oestrous synchronization treatments (84.7, 73.6 and 78.9, groups 1–3 respectively, p < 0.001). These findings suggest that measurement of VER may improve accuracy of oestrus diagnoses when selecting cattle for AI following oestrous synchronization programmes involving tropically adapted cattle.     

Risk factors for and reproductive outcomes of phantom cows on New Zealand dairy farms

AIMS: To determine some of the risk factors for cows not observed in oestrus within 35–42 days of an unsuccessful artificial insemination (AI; phantom cows), and the reproductive outcomes and effect of treatment of phantom cows.

MATERIALS AND METHODS: Over 2 years, in dairy herds from the Waikato (n=10) and Canterbury (n=4) regions of New Zealand, pregnancy diagnosis was carried out 35–42 days after AI on cows that had been inseminated in the first 3 weeks after the start of mating (PSM) but had not been seen returning to oestrus. Risk factors for phantom cows were analysed using a generalised linear mixed effect model.

In Year 1, all phantom cows were left untreated. In Year 2, phantom cows were categorised as having a corpus luteum (CL) (CL+ n=120), or having ovarian follicles ≥10 (n=101) or <10 (n=40)?mm in diameter. Cows with a CL were treated with cloprostenol or untreated and placed with bulls. Cows with no CL received intravaginal progesterone (P4) for 7 days, with injection of gonadotrophin-releasing hormone (GnRH) on Days 0 and 9, and cloprostenol on Day 7 followed by AI. Pregnancy diagnosis of all cows took place 100–120 days after PSM and interval to conception and final pregnancy rate determined.

RESULTS: Overall, of cows inseminated in the first 3 weeks after PSM that did not return to oestrus, 610/6,734 (9.1%) were phantom cows. From the final multivariable analysis, treatment for anoestrus, BCS ≤4.0 at mating, being 2 or >6 years of age, and pure-bred, and decreasing interval between calving and mating, until 98 days post calving, were associated with increased odds of being a phantom cow. Compared to all other groups of cows, phantom cows had a longer interval to conception (p<0.001) and a lower final pregnancy rate (p<0.001).

Treatment of CL+ cows or cows with follicles ≥10?mm did not affect reproductive outcomes (p>0.3). For cows with follicles <10?mm treatment decreased the final percentage not pregnant (3/27; 11%; p=0.01) and interval to conception (21 days; p=0.02) compared with controls (7/13; 54% and 37 days, respectively).

CONCLUSIONS AND CLINICAL RELEVANCE: Risk factors for phantom cows were identified that could be manipulated to reduce the number of phantom cows in a herd, in particular increasing BCS. Treatment of the majority of phantom cows did not improve reproductive performance.     

Impact of Buserelin Acetate or hCG Administration on the Day of First Artificial Insemination on Subsequent Luteal Profile and Conception Rate in Murrah Buffalo (Bubalus bubalis)

This study was designed to investigate the impact of buserelin acetate (BA) or human chorionic gonadotropin (hCG) administration on the day of first artificial insemination (AI) on subsequent luteal profile (diameter of corpus luteum (CL) and plasma progesterone) and conception rate in Murrah buffalo. The present experiment was carried out at two locations in 117 buffalo that were oestrus‐synchronized using cloprostenol (500 μg) administered (i.m.) 11 days apart followed by AI during standing oestrus. Based on treatment (i.m.) at the time of AI, buffalo were randomly categorized (n = 39 in each group) into control (isotonic saline solution, 5 ml), dAI‐BA (buserelin acetate, 20 μg) and dAI‐hCG (hCG, 3000 IU) group. Out of these, 14 buffalo of each group were subjected to ovarian ultrasonography on the day of oestrus to monitor the preovulatory follicle and on days 5, 12, 16 and 21 post‐ovulation to monitor CL diameter. On the day of each sonography, jugular vein blood samples were collected for the estimation of progesterone concentrations. All the buffalo (n = 117) were confirmed for pregnancy on day 40 post‐ovulation. The conception rate was better (p < 0.05) in dAI‐BA (51.3%) and dAI‐hCG (66.7%) groups as compared to their control counterparts (30.8%). Furthermore, the buffalo of dAI‐hCG group had improved (p < 0.05) luteal profile, whereas the buffalo of dAI‐BA group failed (p > 0.05) to exhibit stimulatory impact of treatment on luteal profile when compared to control group. In brief, buserelin acetate or hCG treatment on the day of first AI leads to an increase in conception rate; however, an appreciable impact on post‐ovulation luteal profile was observed only in hCG‐treated Murrah buffalo.     

Role of the sensitivity of detection of oestrus in the submission rate of cows treated to resynchronise oestrus

OBJECTIVE: To determine if failure to detect oestrus in cows treated to resynchronise oestrus leads to fewer cows being inseminated than are truly in oestrus. PROCEDURE: Cows in three herds were enrolled in a controlled breeding program that involved synchronisation of oestrus for a first round of artificial insemination (AI) followed by resynchronisation for a second round of AI. Just before oestrus was expected at the second round of AI, aids for the detection of oestrus were fitted, which included pedometers, radiotelemetric transmitters (HeatWatch), tail-paint and heatmount detectors. Milk samples were collected at the second round of AI (day 33, herds A and B; day 35, herd C of the treatment program) and were used in combination with pregnancy testing to determine the number of cows that were in oestrus (milk progesterone < 2.0 ng/mL) and cows that were not in oestrus (milk progesterone > 2.0 ng/mL or pregnant at second round of AI) at the time samples were collected. RESULTS: The mean sensitivity of detection of oestrus at the resynchronised oestrus was 92.5% and did not differ significantly between herds (P = 0.19). A total of 75% (60/80) of cows that were retrospectively determined to be not pregnant at the time of the second round of AI were classified as having high (> or = 2.0 ng/mL) concentrations of progesterone in milk at that time. Pregnancy testing of cows about 35 days after AI suggested that early pregnancy loss also contributed to a reduction in submission rates at the resynchronised oestrus. CONCLUSION: Failure to submit cows for insemination at a resynchronised oestrus was mainly due to cows not being in oestrus rather than due to a failure to detect oestrus.     

Follicle Turnover and Pregnancy Rates Following Oestrus Synchronization Protocols in Mediterranean Italian Buffaloes (Bubalus bubalis)

An ultrasound assessment of follicle turnover following two different protocols for synchronization of oestrus and ovulation, as well as an assessment of achieved synchronization between ovulation and AI and conception rates in nulliparous and pluriparous buffaloes were carried out during months of increasing day length. Nulliparous buffaloes (n = 30) were subjected only to Ovsynch protocol whereas pluriparous buffaloes (n = 31) were assigned to Ovsynch (n = 14) or to PRID-pregnant mare serum gonadotrophin (PMSG) (n = 17) protocol according to the presence of functional CL confirming cyclic and acyclic conditions. Ultrasound examination of ovarian follicular dynamics at critical days in the course of synchronization treatments was employed to monitor the fate of the largest available follicles at the beginning of treatments. Such available dominant follicle would persist throughout the protocol as ovulating follicle (no-follicle shift) or would regress giving way to a new follicle to become dominant and ovulate (follicle shift). Furthermore, ultrasound monitoring would determine the degree of synchronization of ovulation and final outcome represented by pregnancy rates. Pregnancy rate following Ovsynch protocol was 40% (12/30) and 42.8% (6/14) in nulliparous and pluriparous buffaloes respectively (p = 0.8575). Most ovulations were synchronized and recorded at AI and the following day in nulliparous (24/30; 80%) and pluriparous (12/14; 85.7%) buffaloes respectively (p = 1.000). A follicle shift was recorded in 14 of 30 (46.6%) and 11 of 14 (78.5%) in nulliparous and pluriparous buffaloes respectively (p = 0.0466). Among established pregnancies: eight derived from follicle shift (66.6%) and four from no-follicle shift (33.3%) in nulliparous buffaloes, p = 0.0729 whereas in pluriparous buffaloes five (83.3%) derived from follicle shift and one from no-follicle shift (16.6%), p = 0.6154. Collectively, from 18 pregnancies in nulliparous and pluriparous buffaloes following Ovsynch protocol, 13 derived from follicle shift (72.2%) and five from no-follicle shift (27.7%), p = 0.0860. Pregnancy rate in pluriparous buffaloes following PRID-PMSG protocol was 70.5% (12/17). The majority of ovulations were synchronized and recorded at first, second AI and following day (13/17; 76.4%). A follicle shift occurred in 15/17 buffaloes (88.2%) and among the 12 recorded pregnancies, 11 derived from follicle shift (91.6%), p = 0.5147. In conclusion, pregnancy rates following Ovsynch protocol were similar in nulliparous and pluriparous cyclic buffaloes. A progestagen treatment on acyclic buffaloes but still displaying some ovarian follicular dynamics, resulted in significantly higher pregnancy rate compared with Ovsynch (p = 0.0376). According to the time of scheduled AI, a high degree of synchronized ovulations were recorded following the implementation of both protocols.     

Evaluation of human chorionic gonadotropin as a replacement for gonadotropin-releasing hormone in ovulation-synchronization protocols before fixed timed artificial insemination in beef cattle

Two experiments were conducted during 2 yr to evaluate differences in ovulation potential and fertility in response to GnRH or hCG. In Exp. 1, 46 beef cows were given 100 microg of GnRH or 500, 1,000, 2,000, or 3,000 IU of hCG. Ovulation incidence was not different between GnRH and any of the hCG doses, indicating that ovulatory capacity of at least 500 IU of hCG was equivalent to GnRH. In Exp. 2, beef cows (n = 676) at 6 locations were assigned randomly to a 2 x 3 factorial arrangement of treatments. Main effects were: 1) pre-timed AI (TAI) treatment (GnRH or hCG) and 2) post-TAI treatment (saline, GnRH, or hCG) to initiate resynchronization of ovulation in previously inseminated cattle. Blood samples were collected (d -21 and -10) to determine progesterone concentrations and assess cyclicity. Cattle were treated with a progesterone insert on d -10 and with 100 microg of GnRH or 1,000 IU of hCG. A PGF(2alpha) injection was given at insert removal on d -3. Cows were inseminated 62 h (d 0) after insert removal. On d 26 after first TAI, cows of unknown pregnancy status were treated with saline, GnRH, or hCG to initiate a CO-Synch protocol. Pregnancy was diagnosed 33 d after first TAI to determine pregnancies per AI (P/AI). Nonpregnant cows at 6 locations in yr 1 and 1 location in yr 2 were given PGF(2alpha) and inseminated 56 h later, concurrent with a GnRH injection. Five weeks later, pregnancy diagnosis was conducted to determine pregnancy loss after first TAI and pregnancy outcome of the second TAI. Injection of pre-TAI hCG reduced (P < 0.001) P/AI compared with GnRH, with a greater reduction in cycling cows. Post-TAI treatments had no negative effect on P/AI resulting from the first TAI. Serum progesterone was greater (P = 0.06) 7 d after pre-TAI hCG than after GnRH and greater (P < 0.05) after post-TAI hCG on d 26 compared with saline 7 d after treatment in association with greater frequency of multiple corpora lutea. Compared with saline, injections of post-TAI GnRH and hCG did not increase second insemination P/AI, and inconsistent results were detected among locations. Use of hCG in lieu of GnRH is contraindicated in a CO-Synch + progesterone insert protocol. Compared with a breeding season having only 1 TAI and longer exposure to cleanup bulls, total breeding season pregnancy rate was reduced by one-third, subsequent calving distribution was altered, and 50% more AI-sired calves were obtained by applying 2 TAI during the breeding season.     

Radioimmunoassay of milk progesterone as a tool for fertility control in smallholder dairy farms

This study focused on the use of radioimmunoassay of progesterone in milk for the diagnosis of post-partum ovarian cyclicity and accurate detection of oestrus and non-pregnancy in cows in the artificial insemination (AI) programme in Bangladesh. In Investigation 1, milk samples were collected on day 0 (day of AI), day 9–13 and day 21–24 from 444 milking cows of various breeds presented for the first post-partum insemination by 413 farmers living at 182 villages/regions in Mymensingh District from 6 AI centres and sub-centres. Each cow was then examined three times after each AI until it stopped returning to oestrus. Sixty to 90 days after the last AI, the cows were examined per rectum to confirm the pregnancy. Milk progesterone data on day 21–24 contributed to a clear diagnosis with respect to non-pregnancy in 100% cows, indicating a possible use of this progesterone assay for identifying non-pregnant cows in AI programmes. In Investigation 2, milk progesterone was monitored two times in a month with a 10-day interval in 88 cows. The samples were taken between 10 days after calving and the first detected oestrus, followed by two more samples 10 days apart. The proportion of cows accurately detected in oestrus was 30%. Another 30% were stated to be in oestrus when they were not (false positive) and 40% were not detected when they were in oestrus (false negative). The mean intervals between calving and oestrus and between calving luteal activity were 40 to 362 days (median = 120, n = 82) and 34 to 398 (median = 111, n = 64) days, respectively. The body condition scores at calving and at the initiation of luteal activity influenced the interval between calving and luteal activity (p < 0.05). Cows suckled twice daily initiated luteal activity earlier than their counterparts suckled several times daily (p < 0.05). Determination of progesterone in milk on day 21–24 is a good means for detecting non-pregnant cows.     

Intravaginal progesterone devices in synchronization protocols for artificial insemination in beef heifers

Two experiments were designed to investigate the administration of intravaginal progesterone in protocols for oestrus and ovulation synchronization in beef heifers. In Experiment 1, cyclic Black Angus heifers (n = 20) received an Ovsynch protocol and were randomly assigned to receive (CIDR‐Ovsynch) or not (Ovsynch) a progesterone device between Days 0 and 7. Treatment with a controlled internal drug release (CIDR) device significantly increased the size of the dominant follicle prior to ovulation (12.8 ± 0.4 CIDR‐Ovsynch vs 11.4 ± 0.4 Ovsynch) (p < 0.02). Plasma progesterone concentrations throughout the experiment were affected by the interaction between group and day effects (p < 0.004). In Experiment 2, cyclic Polled Hereford heifers (n = 382) were randomly assigned to one of the six treatment groups (3 × 2 factorial design) to receive a CIDR, a used bovine intravaginal device (DIB), or a medroxiprogesterone acetate (MAP) sponge and GnRH analogues (lecirelin or buserelin). All heifers received oestradiol benzoate plus one of the devices on Day 0 and PGF on Day 7 pm (device withdrawal). Heifers were detected in oestrus 36 h after PGF and inseminated 8–12 h later, while the remainder received GnRH 48 h after PGF and were inseminated on Day 10 (60 h). The number of heifers detected in oestrus on Day 8 and conception rate to AI on Day 9 were higher (p < 0.01) in the used‐DIB than in the CIDR or MAP groups, while the opposite occurred with the pregnancy rate to FTAI on Day 10 (p < 0.01). There was no effect of progesterone source, GnRH analogue or their interaction on overall pregnancy rates (64.9%). Progesterone treatment of heifers during an Ovsynch protocol resulted in a larger pre‐ovulatory follicle in beef heifers. Progesterone content of intravaginal devices in synchronization protocols is important for the timing of AI, as the use of low‐progesterone devices can shorten the interval to oestrus.     

Comparison of a camera - software system and typical farm management for detecting oestrus in dairy cattle at pasture

AIM: To compare the sensitivity, specificity, predictive values and accuracy of detection of oestrus using a novel oestrus detection strip (ODS) and a camera-software device (CSD) with typical farm management practices of visual observation and use of tail paint in dairy cattle at pasture. METHODS: Dairy cows (n = 480) in a seasonal-calving herd managed at pasture under typical commercial conditions in New Zealand were stratified by age, body condition score and days in milk, then randomly allocated to one of two groups prior to the planned start of mating (PSM). Tail paint was applied to all cows and oestrus detected by visual observation of oestrous behaviour and removal of paint, by farm staff. One group (n = 240) was fitted with ODS and also monitored for signs of oestrus using a CSD, while the Control group (n = 240) was monitored using tail paint and visual observations only. Cows detected in oestrus were artificially inseminated (AI), and pregnancy status determined using rectal palpation and ultrasonography, 51-52 days after the end of a 55-day A period. Results of pregnancy diagnosis were used to confirm the occurrence of oestrus, and the sensitivity, specificity, predictive value and accuracy of detection of oestrus compared between oestrus detection methods. RESULTS: The sensitivity and accuracy of oestrus detection in the Control group, using visual observation and tail paint, were low. Compared with the Control group, detection of oestrus using the ODS and CSD resulted in greater sensitivity (85% vs 78%; p = 0.006), specificity (99.6% vs 98.0%; p < 0.001), positive predictive value (PPV; 88% vs 51%; p < 0.001) and overall accuracy (99.0% vs 98.0%; p < 0.001). Negative predictive value (NPV) did not differ significantly between groups (99.4% vs 99.3%; p = 0.28). Pregnancy rate to first service was higher in the CSD group than in the Control group (72% vs 39%; p < 0.05). Use of the CSD significantly increased the cumulative proportion of cows pregnant to AI over the breeding period (p = 0.044). CONCLUSION AND CLINICAL RELEVANCE: The ODS and CSD was satisfactory for detection of oestrus in seasonal calving dairy herds grazing on pasture and could improve the sensitivity and accuracy of detection of oestrus in herds where these are low.     

Field experiences with early pregnancy diagnosis by progesterone-based ELISA in sows

In four Kenyan pig breeding units the pregnancy diagnosis of sows has been carried out in two groups: Group 1 (n = 1911): the sows were transrectaly pregnancy tested between Days 17-22 post-mating by ultrasound. Sows testing non-pregnant immediately received one dose of 400 IU pregnant mare serum gonadotropin (PMSG) (equine chorion gonadotropin, eCG) and 200 IU human chorion gonadotropin (hCG). On showing signs of oestrous, the animals were subsequently artificially inseminated (AI). Group 2 (n = 1923): sows were pregnancy tested by serum progesterone (P4)-based enzyme-linked immunosorbent assay (ELISA) on Day 17 post-breeding. P4 concentrations were categorized as positive (> 5 ng/ml) or negative (< 5 ng/ml). Sows testing nonpregnant immediately received one dose of 400 IU PMSG and 200 IU hCG by injection, and were subsequently artificially inseminated. The following parameters were evaluated: sows diagnosed non-pregnant, days from first post-weaning insemination until the sows were inseminated at their first return to oestrus; farrowing rate and total piglets born and number of live-born piglets in litters. The percentage of sows diagnosed non-pregnant in the two groups, as well as the totals of born piglets and of live-born piglets in litters did not differ significantly between the two groups. The number of days from the first post-weaning mating until the sows were artificially inseminated at their first return to oestrus and the administration of eCG and hCG was shorter (P < 0.01) and farrowing rate was higher (P< 0.01) in the ELISA-tested sows.     

Treatment of Unobserved Oestrus in a Dairy Cattle Herd with Low Oestrous Detection Rate up to 60 days Post-partum

The efficiency of treatments for unobserved oestrus and their effect on the reproductive performance of a dairy cattle herd with low oestrous detection rate till 60 days post‐partum (dpp), attributed to the declivous and slippery concrete floor were investigated. The herdsman requested advice in order to improve the mean days open of the herd, but no investments were allowed because a new unit was about to be built. Due to the low oestrus detection rate of the herd, the breeding policy was to inseminate at the first detected post‐partum oestrus. Cows were examined at 20–30 dpp to assess uterine involution, ovarian activity and prevalence of reproductive disorders and, at 60 dpp if no previous oestrus was detected. Each examination included palpation per rectum, ultrasound scanning and collection of a blood sample for plasma progesterone (P4) measurement. Cows with unobserved oestrus till 60 dpp were allocated either to a treatment group (n=139) or to a control group (n=139). Three treatments were used: (a) injection of PGF_2α (PG) upon detection of a corpus luteum (CL; n = 30), cows not observed in oestrus being re‐injected 11–12 days later. AI was at oestrus; (b) PRID (n=35) or Crestar (n=74) devices kept in situ for 12 and 9 days, respectively, were associated to an injection of PG and of equine chorionic gonadotrophin (eCG) at device removal. Cows were double‐fixed time‐inseminated at 48 and 72 h after device removal. All treated cows were examined at 48–72 h after treatment to confirm oestrus. The percentage of cows detected in oestrus up to 60 dpp remained unchanged through the trial (35 and 47% for years before intervention: 1994–95; 51 and 48% for years of intervention: 1996–97). In contrast, the oestrous detection rate was high both in treated (93%) and control (100%) cows. This possibly resulted from an improvement in the oestrous detection efficiency of the herd's personnel and from examination of cows at 48–72 h after treatment. Treated and control cows had identical conception rate (CR; 36 and 37%, respectively) and reproductive performance. However, the mean days open of the herd in 1996 was significantly improved in comparison with previous years (mean ± SEM: 134 ± 6, 126 ± 5, 110 ± 4 and 114 ± 5 days, for years 1994, 1995, 1996 and 1997, respectively, p < 0.05, ANOVA ). Conception rate to AI up to 40 dpp was significantly reduced, compared with the period between 60 and 100 dpp but, mean days open were significantly improved in cows inseminated up to 60 dpp, compared with thereafter (p < 0.05).     

Effect of Complex Vertebral Malformation on Luteal Function in Holstein Cows During Oestrous Cycle and Early Pregnancy

The reason why cows carrying the mutation of complex vertebral malformation (CVM) show poor reproductive capability although they carry only one mutant allele is still not fully understood. Monitoring the progesterone profiles during oestrous cycle and early pregnancy in carrier cows might help explain their lowered reproductive capability. Progesterone concentration was measured in 19 CVM carrier cows and 21 control cows during oestrous cycle and early pregnancy. Milk samples were collected from all cows starting on the day of artificial insemination until day 45 post‐AI. Progesterone was measured in skim milk using enzyme‐linked immunosorbent assay (ELISA). Progesterone concentration was significantly reduced on day 7 (p < 0.05) and day 9 (p < 0.01) post‐insemination in conceived CVM carrier cows when compared with that in control conceived cows. The mean progesterone concentration during early pregnancy was significantly lower (p < 0.05) in conceived cows with CVM than that of control cows in the same period. However, the mean progesterone concentration did not differ significantly (p = 0.072) in CVM cows that showed fertilization failure or embryonic death than that of control cows. Additionally, of 13 conceived control cows, eight cows (61.5%) showed normal luteal function. In contrast, of nine conceived CVM cows, only four cows (44.4%) showed normal luteal function. The conception rate was 47.4% in CVM carrier cows and 61.9% in control cows, but this difference did not reach significance. In conclusion, progesterone concentration might be lowered during early pregnancy in conceived CVM cows compared with that in control cows.     

Shortening the Postpartum Anoestrous Interval in Suckled Crossbred Dual Purpose Cows Using Progestagen Intravaginal Sponges plus eCG and PGF2α

One hundred and twenty‐six suckled crossbred cows (Bos taurus × Bos indicus), with body condition score ≥3 (1–5 point scale), were employed in the present study to evaluate the effectiveness of intravaginal progestin‐releasing sponges (IVS) for shortening anoestrous interval. Fifty‐four cows were assigned to control group. Seventy‐two cows were treated with IVS impregnated with 250 mg of medroxy‐acetate‐progesterone (MAP) as follows: day 0, IVS plus 5 mg of 17β‐E and 50 mg of MAP i.m.; day 6, 500 IU of equine chorionic gonadotropin and 25 mg prostaglandin F_2α i.m.; day 8, IVS withdrawal and day 9, 1 mg 17β‐E i.m. Cows were also grouped according to postpartum days (dpp) at treatment: MAP <70 days (n = 25); control <70 days (n = 22); MAP >70 days (n = 47); control >70 days (n = 32). From IVS removal, cows were detected in oestrus and inseminated. Cows not detected in oestrus were timed artificial insemination 72 h after sponge removal. Treatment effect on oestrous rate (ER), conception rate (CR), pregnancy rate (PR) and treatment to conception intervals (TCI) and calving to conception intervals (CCI) were evaluated. The ER, CR and PR were analysed using proc logistic , while TCI and CCI with proc glm of SAS. The groups MAP <70 days and MAP >70 days showed higher (p < 0.01) ER than control <70 days and control >70 days (84.0% and 76.6% vs 31.8% and 31.3% respectively). The PR was higher (p < 0.01) in MAP <70 days vs control <70 days (64.0% vs 22.7%) and also higher (p < 0.05) in MAP >70 days vs control <70 days (40.4% vs 18.8%). The TCI and CCI were shorter (p < 0.01) in MAP <70 days vs control <70 days (36.0 and 95.8 days; 95.3 and 158.6 days respectively). In conclusion, only cows treated with IVS before 70 dpp had a CCI shorter than 100 days, consequently this treatment shortened postpartum anoestrous interval in crossbred dual purpose cattle.     

hCG‐Induced Ovulation in Thoroughbred Mares Does Not Affect Corpus luteum Development and Function During Early Pregnancy

Our aim was to compare Corpus luteum (CL) development and blood plasma concentration of progesterone ([P₄]) in thoroughbred mares after spontaneous (Control: C) or human chorionic gonadotrophin (hCG)‐induced ovulation. Lactating mares (C = 12; hCG = 21) were daily teased and mated during second oestrus post‐partum. Treated mares received 2500 IU hCG i.v. at first day of behavioural oestrus when dominant follicular size was >35, ≤42 mm and mated 12–24 h after. Control mares in oestrus were mated with dominant follicular size ≥45 mm. Dominant follicle before ovulation, CL and gestational sac were measured by ultrasound and [P₄] by radioimmunoassay (RIA). Blood sampling and ultrasound CL exams were done at days 1, 2, 3, 4, 8, 12, 16, 20, 25, 30, 35, 40, 45, 60 and 90 after ovulation and gestational sac from day 12 after ovulation in pregnant (P) mares; non‐pregnant (NP) were followed until oestrus returned. Data analyses considered four subgroups: hCG‐P, hCG‐NP, C‐P and C‐NP. Preovulatory follicular size was smaller in hCG mares than in C: 39.2 ± 2.7 mm vs 51.0 ± 1.8 mm (p < 0.0001). All hCG mares ovulated 24–48 h after treatment and presented similar oestrus duration as controls. C. luteum size in P mares showed the same pattern of development through days 4–35, presenting erratic differences during initial establishment. Thus, on days 1 and 3, CL was smaller in hCG‐P (p < 0.05); while in hCG‐NP, CL size was greater than in C‐NP on day three (p = 0.03). Corpus luteum size remained stable until day 90 in hCG‐P mares, while in C‐P a transient and apparently not functional increase was detected on days 40 and 45 (p < 0.05) and the decrease from day 60 onwards, made this difference to disappear. No differences were observed in [P₄] pattern between P, or between NP subgroups, respectively. So, hCG‐induced ovulation does not affect CL development, neither [P₄] during early pregnancy. One cycle pregnancy rate tended to be lower in hCG mares while season pregnancy rates were similar to controls.     

Pregnancy rate after timed artificial insemination in early post-partum dairy cows after Ovsynch or specific synchronization protocols

The present study was designed to compare the reproductive performance of pre-synchronized post-partum dairy cows subjected, either to the Ovsynch protocol without screening for ovarian status (control group), or to a specific oestrous synchronization protocol applied according to their ovarian status, as determined by transrectal ultrasound (experimental group). The study was conducted on 428 lactating dairy cows. Cows in the Ovsynch group (n = 205) were synchronized and time inseminated after receiving the Ovsynch protocol treatment. Cows in the specific synchronization (Ssynch) group (n = 223) were weekly subjected to transrectal ultrasound exams for 4 weeks, or until AI or starting treatment, and divided into four subgroups according to their ovarian status: (i). corpus luteum (CL) subgroup (n = 130), cows with a CL; (ii). natural oestrus (NE) subgroup (n = 58), cows showing NE; (iii). anovulatory follicles (AF) subgroup (n = 26), cows considered to have AF; and (iv). ovarian cysts (OC) subgroup (n = 9), cows with OC. Cows in the Ssynch group were synchronized and time inseminated following a specific oestrous synchronization protocol, or inseminated at NE. Logistic regression analysis was carried out for the dependent variables ovulation and pregnancy rates to first and to second AI (second AI: first AI + return AI). Cows subjected to Ssynch were 2.1 times more likely to become pregnant at first and at second AI compared with those synchronized using the Ovsynch protocol (P < 0.0001). Our results show that the response of post-partum pre-synchronized cows to a specific oestrous synchronization protocol applied according to their ovarian status is more effective than the response to the Ovsynch protocol applied without taking into account the ovarian status of the animals.     

The Effect of Intravaginal Applied GnRH-agonist on the Time of Ovulation and Subsequent Reproductive Performance of Weaned Multiparous Sows

In order to prove the effect of 'fixed time insemination' and insemination at standing oestrus after post-weaning application of GnRH, in a Croatian large breeding unit, 502 sows were assigned to three groups and were artificially inseminated (AI) at their first post-weaning oestrus as many times as they stand, in 24-h intervals. The groups were treated as follows: group 1 (control, n = 160) were AI during their standing reflex; group 2 ['GnRH-fixed time insemination' (GnRH-FT-AI), n = 175] were AI, independent of detection of oestrus and following administration of GnRH-agonist at 96 h post-weaning; group 3 [GnRH insemination at standing oestrus (GnRH-OE-AI), n = 167] the animals were GnRH-agonist treated as group 2 and were AI at their standing reflex. Pre-trial daily average lactational feed intake, average daily feed intake from weaning to oestrus, oestrus within 6 days post-weaning (%), ovulation within 6 days post-weaning (%), weaning-to-oestrus interval (h), duration of oestrus (h), follicle size (mm), interval from oestrus to ovulation (h), subsequent day 24 pregnancy rate (%), farrowing rate (%) and total pigs born were evaluated. Pre-trial average daily lactational voluntary feed intake was 7.1 +/- 0.08 kg in group 1, 7.0 +/- 0.07 kg in group 2 and 7.1 +/- 0.17 kg in group 3 (p > 0.05). Average voluntary daily feed intake from weaning to oestrus was 5.1 +/- 0.3 kg in group 1, 5.2 +/- 0.5 kg in group 2 and 5.2 +/- 0.19 kg in group 3 (p > 0.05). Oestrus was detected within 6 days post-weaning in 134 (83.8%) in control, 164 (93.7%) in GnRH-FT-AI and 155 (92.8%) animals in GnRH-OE-AI groups (p = 0.05). Follicle size did not differ (p > 0.05) among the groups. In control 82.8%, in GnRH-FT-AI 91.5% and in GnRH-OE-AI 91.0% of the sows ovulated within 6 days post-weaning (p = 0.04), and had 80.6, 90.9 and 89.7% 24-day pregnancy rates (p = 0.16), respectively. In GnRH-FT-AI group 90.2%, in GnRH-OE-AI sows 89.7%, in control animals 79.9% farrowing rates were recorded (p = 0.17). Weaning to oestrus interval was 113.1 h in control, 114.1 h in GnRH-FT-AI and 112.6 h GnRH-OE-AI (p > 0.05). Duration of oestrus was significantly shorter in GnRH-FT-AI (44.9 h) and GnRH-OE-AI (48.1 h) animals, compared with the control (62.9 h) sows (p = 0.001). Similarly, the interval from oestrus to ovulation revealed significant (p = 0.004) differences between the groups (control 44.1 h, GnRH-OE-AI 34.1 h and GnRH-FT-AI 32.9 h). GnRH-FT-AI (12.5) and GnRH-OE-AI (12.6) sows had significantly higher (p = 0.01) number of total pigs born (n = 10.4) compared with control sows. GnRH-agonist-gel treatment to the sow shortens duration of oestrus, the interval from oestrus to ovulation, and may eliminate the need for oestrus detection in the hands of skilled personnel.     

Fertility of Holstein cows and heifers submitted to timed artificial insemination and receiving one or two doses (12 h apart) of semen

The objective of this retrospective study was to assess the effect of receiving a single (n = 50,285) or double (n = 4392) artificial insemination (AI), 12 h apart, within a timed artificial insemination protocol on pregnancy per AI (P/AI) in nulliparous heifers (inseminated with either sex-sorted or conventional semen) and pluriparous Holstein cows in a commercial dairy herd. Also, this study aimed to investigate the relationship between temperature-humidity index (THI) and time of the first AI and fertility. Fertility of cows receiving two AI with normothermia (THI <68) was higher (p < .05) than cows receiving a single AI (42.9% vs. 36.4%). P/AI of cows receiving two AI with severe heat stress (THI >85) was higher (p < .05) than cows receiving a single AI (21.0% vs. 12.6%). Regardless of heat stress conditions, applying the first AI in the morning increased (p < .05) P/AI in cows with double AI than in cows whose first AI occurred in the afternoon (38.4 vs. 33.3%). With moderate heat stress, and sexed-sorted semen, P/AI to timed AI was higher (65.0 vs. 51.9%; p < .05) in heifers receiving double AI than those serviced once. It was concluded that double AI, 12 h apart, enhanced fertility at timed AI than herd mates with a single AI, particularly with heat stress at breeding.