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We tested whether utilising the male effect to stimulate ewes before the mating period can reduce the time to conception following the introduction of entire rams, and increase fertility, prolificacy, and reproductive rate (number of fetuses per 100 ewes exposed to fertile rams). A retrospective analysis was used to analyse records from 59,716 ewes collected over 34 years (1986–2020) from seven genotypes: Border Leicester, Composite (crossbred), Dorset, Merino, Dorset x Polypay, Rambouillet, White Suffolk. The dataset also included nulliparous young ewes (mated at age 8 months) and adult parous ewes. Vasectomized rams were used to stimulate 20,632 ewes before a mating period that lasted 2 or 3 estrous cycles, and the outcomes were compared with those from 39,084 ewes that had not been stimulated. Independently of genotype, utilising the male stimulus advanced the average conception date by 8 days for young ewes (P < 0.0001) and by 1 day for adult ewes (P < 0.0001). The male stimulus also increased the proportion of ewes that conceived in their first cycle by 33 % for young ewes and by 6 % for adult ewes (P < 0.0001). For the cycle of conception, there were significant (P < 0.0001) effects of two interactions: male stimulus x age at mating and male stimulus x live weight at mating. The male stimulus improved fertility in both adult ewes (99.8 % vs 89 %; P < 0.001) and young ewes (77.7 % vs 81.3 %; P < 0.001). The male stimulus increased the number of young ewes (41.9 % vs 11.1 %; P < 0.001) and adult ewes (16.6 % vs 2.7 %; P < 0.001) that conceived multiple fetuses in the first 17 days of the mating period. The reproductive rate was improved by the male stimulus in young ewes (129 % vs 135 %; P < 0.001) but not in adult ewes (120 % vs 122 %; P = 0.12). When all animals for all breeds were included in the analyses, there were improvements in fertility, prolificacy, and reproductive rate as age and live weight increased at mating (P < 0.0001). We conclude that, independently of genotype, utilising the male stimulus before the mating period reduces the time to conception and improves reproductive performance in both young and adult ewes. • Male stimulus advanced the average conception date for ewe lambs and adult ewes. • Male stimulus increased the proportion of ewes conceived in their first reproductive cycle. • Male stimulus improved fertility in both adult ewes and ewe lambs. • Male stimulus increased the number of ewes that conceived multiple foetuses in the first 17 days of the mating period. • Male stimulus improved the reproductive rate in ewe lambs. [ABSTRACT FROM AUTHOR]

The objective of this study was to create a stochastic, agent-based simulation model of a synthetic population of beef cattle, and then use it to compare the technical performance of different reproductive strategies. The model was parameterized using data from a real beef cattle herd and from the peer-reviewed scientific literature to represent a Nelore cattle herd in the state of São Paulo, Brazil. Ten scenarios were evaluated: natural mating (NM) only (ONM); one timed artificial insemination (TAI) plus NM (1TAI + NM); two TAI plus NM, with 24, 32, and 40 days between inseminations (2TAI/24 + NM, 2TAI/32 + NM, and 2TAI/40 + NM, respectively); three TAI without NM, with 24, 32, and 40 days between TAI (3TAI/24, 3TAI/32, and 3TAI/40, respectively); and three TAI plus NM, with 24 and 32 days (3TAI/24 + NM and 3TAI/32 + NM, respectively). NM began 10 days after the last TAI and was performed until the end of the breeding season. The size of the female herd was set to contain up to 400 individuals. The bull population was established at 0, 7, or 15 bulls depending on the used scenario. Simulation was performed for 5000 days. The outcomes for each scenario are means ± S.E. assessed on 32 farms at 1-day time intervals and on an animal-by-animal basis after steady state was reached (1825 days). The 3TAI/24 + NM scenario resulted in a greater number of births (279.85 ± 0.47 births), while the ONM scenario had the least value (202.38 ± 0.43 births). The heaviest males and females at weaning belonged to 3TAI/24, with 190.85 ± 0.17 kg for males and 173.89 ± 0.13 kg for females. The ONM scenario had the lightest males (166.84 ± 0.18 kg) and females (151.75 ± 0.16 kg). The greatest and least total pregnancy rates were found in 3TAI/24 + NM (0.91 ± 0.00) and ONM (0.62 ± 0.00), respectively. The ONM scenario required 52.5 days more than scenarios that included TAI to reach 50% of pregnancy. The greatest ages at culling for cows was 3TAI/24 + NM (3658.88 ± 10.41 days). In contrast, the lowest age at culling was found in ONM (2823.93 ± 8.28 days). We concluded that the proposed model represents the main interactions of a real beef cattle herd. It has all the advantages of a physical experiment, but does not require incurring significant expenses nor altering the real system. This study offers evidence that the scenarios that present the best technical performance are those that used TAI with a 24-day interval between inseminations. • A stochastic agent-base model was created to simulate a cow-calf operation. • Performance of using natural mating alone or in combination with TAI was evaluated. • Number of TAI (1, 2 or 3) and days between them (24, 32 and 40) were evaluated. • All TAI scenarios showed better reproductive performance and weaning weights. • 3 TAI plus natural mating (24 days of interval) presented the best technical results. [ABSTRACT FROM AUTHOR]

To meet growing worldwide demands for animal products, animal production will need to increase in capacity and efficiency. Every opportunity to improve animal protein yield should be considered and explored. Developmental programming is one such opportunity that has not yet been thoroughly investigated in farm animal production. While developmental programming can be advantageous for the survival of the offspring, it is often described in conjunction with negative consequences. The known and potential causes and mechanisms are numerous, often stemming from some sort of stress experienced during the prenatal or early postnatal period. One stressor that is particularly concerning for farm animal production is heat stress. Heat stress is known to elicit adaptations associated with developmental programming in several species, but has not been investigated in dairy cattle until recently. Multiple studies have shown that heat stress experienced during the periconceptional period is generally associated with reduced milk production of resulting offspring. This could be the result of adaptations within the pre-ovulatory oocyte or early developing embryo. Interestingly, in a few select comparisons, periconceptional heat stress was associated with greater milk production. This was only observed when dairy cattle calved in the spring, and would therefore be reaching peak milk production in late spring or early summer (in heat stress). This is consistent with the match/mismatch theory associated with developmental programming, where matched prenatal/postnatal environments confer advantageous adaptations and mismatched prenatal/postnatal environments are generally detrimental to the offspring. While these studies are important additions to our growing knowledge of heat stress impacts on dairy cow production, the broader implication of developmental programming requires further investigation. [ABSTRACT FROM AUTHOR]

The objectives of this study were to (1) characterize the variability for sire's predicted transmitting ability (PTA) for daughter's pregnancy rate (DPR) and cow conception rate (CCR) , (2) determine the relationships among sire's PTA for DPR, sire's PTA for CCR, daughter's parity, and daughter's 305-d mature-equivalent (ME) milk production, and (3) evaluate the associations among sire's PTA for DPR and CCR and daughter's reproductive performance [pregnancy to first artificial insemination (P/AI) , pregnancy by 150 d in milk (PR150) and pregnancy loss (PL) after first AI] in Canadian Holstein cows. The data were obtained from 822 lactating Holstein cows from 10 commercial dairy herds located in Alberta. Overall mean (range) for sire's PTA for DPR and CCR was −0.09 (−9.6 to 8.2) and −0.05 (−9.9 to 7.4), respectively. Sire's PTA for DPR was strongly and positively associated with sire's PTA for CCR (r = 0.89; P < 0.01). Sire's PTA for DPR and CCR were weakly and negatively associated with parity (r = −0.15 and −0.17, respectively; P < 0.01); however, the amount of phenotypic variation in sire's PTA for DPR and CCR explainable by parity was small (coefficient of determination; R2 = 0.02). The daughter's 305-d ME milk production tended to be weakly but positively associated with sire's PTA for DPR (r = 0.06; P = 0.07) and CCR (r = 0.06; P = 0.10); however, the amount of phenotypic variation in daughter's 305-d ME milk production explainable by changes in sire's PTA for DPR or CCR was small (R2 = 0.004 and 0.003, respectively). Overall P/AI, PR150 and PL were 38, 65 and 12%, respectively. Sire's PTA for DPR tended to be associated with P/AI (P = 0.08) but was significantly associated with PR150 (P = 0.03). For every unit increase in sire's PTA for DPR, the estimated probabilities for P/AI and PR150 increased by 6.2 and 7.5%, respectively. Sire's PTA for CCR was not associated with P/AI or PR150. Neither sire's PTA for DPR nor CCR were associated with PL. In summary, sire's PTA for DPR and CCR were highly variable and positively correlated. Sire's PTA for DPR was associated with daughter's P/AI and PR150, but sire's PTA for CCR was not associated with any of the measured reproductive outcomes. Therefore, selecting sires with high PTA for DPR has the potential to improve the reproductive efficiency of Canadian dairy herds. • The associations among sire's predicted transmitting ability (PTA) for daughter's pregnancy rate (DPR) and cow conception rate (CCR) and daughter's reproductive performance were evaluated. • Sire's PTA for DPR and CCR were strongly correlated (r = 0.89; P < 0.01). • Overall mean (range) for sire's PTA for DPR and CCR was −0.09 (−9.6 to 8.2) and −0.05 (−9.9 to 7.4), respectively. • Sire's PTA for DPR tended (P = 0.08) to be associated with pregnancy to first AI and was significantly (P = 0.03) associated with pregnancy by 150 days in milk. • Neither sire's PTA for DPR nor CCR were associated with pregnancy loss. [ABSTRACT FROM AUTHOR]

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

The objective of this study was to improve prediction of the probability of conception in dairy cows with clinical endometritis by means of a combination of examination results. For this purpose, a total of 1386 dairy cows were screened for clinical endometritis and data from 286 cows were collected including the calving history, puerperal or concurrent diseases and cow-specific data (breed, parity, BCS at puerperal control 1 (PC 1 = 22–45 days in milk (DIM)), and daily milk yield). The diagnosis of clinical endometritis was made at PC 1 by vaginal discharge scoring with a Metricheck device on a scale from 1 to 3 (endometritis score (ES) 1 to 3). A detailed clinical and gynecological examination including external inspection, transrectal palpation and ultrasonographic examination of the genital tract (cervix, uterus, and ovaries), vaginoscopy and microbiological analysis of the intrauterine content were performed. All cows with clinical endometritis (n = 286) were treated according to the ovarian findings (corpus luteum, dominant follicle) at PC 1. A second puerperal control was performed 21 ± 1 days later. Data was collected until 200 DIM. The risk factors for ES 3 and their effect on the distribution of ES were determined by the Chi-square test and binary logistic regression. Multivariable implications of a large sum of examination parameters on the pregnancy outcome at different DIM (100, 150 and 200 DIM) were analyzed on the basis of conditional inference trees. The prevalence of clinical endometritis was 28%. The ovarian findings at PC 1 did not have any impact on reproductive performance. Puerperal diseases prior to the study, visible vaginal discharge (VVD) and the presence of Trueperella pyogenes (TP) were associated with an ES 3. The prediction of pregnancy status for all DIM times was dominated significantly by VVD or TP as decisive factors. Cows with VVD or TP had lower conception rates at all DIM times than cows without these findings, but the predictive accuracy was similar for both groups. These results suggest that VVD is a valuable and practical examination parameter, which can be used for pregnancy prediction on farm. Therefore, it might facilitate early breeding decisions in cows with clinical endometritis under farm conditions. • Visible vaginal discharge and T. pyogenes are decisive for pregnancy prediction. • These factors are associated with low conception rates at 100, 150 and 200 DIM. • The predictability of pregnancy is similar for both factors. • Visible vaginal discharge can replace microbiology for breeding decisions. • Ultrasound findings do not improve the predictability of the pregnancy outcome. [ABSTRACT FROM AUTHOR]

Conception rate with the use of sexed semen is lower than that with the use of conventional semen, posing a major problem in the dairy industry. The aim of this study was to understand the risk factors that affect the conception rate after artificial insemination (AI) with conventional and sexed semen using field data. The records of the first insemination in Holstein heifers with conventional (n = 41,857) and sexed semen (n = 45,465) in Hokkaido, Japan were analyzed. The mean conception rate after AI from 2012 to 2016 was 56.9% with conventional semen and 47.3% with sexed semen. A multivariable logistic regression model including the effects of year, heifer age, time of the year, semen type, service sire, and their interactions was used to evaluate the interaction effect of heifer age and time of the year by semen type on the conception rate. In the analysis using heifer age, we found that heifers inseminated with sexed semen were approximately 21 days younger than those inseminated with conventional semen. Interestingly, in early, warmer months (Jun, Jul, and Aug), the conception rate after AI with sexed semen significantly decreased compared with that after AI with conventional semen (P < 0.01). Our results showed that more careful implementation of AI is required for a stable conception using sexed semen, particularly during warmer months. • The use of sexed semen increased as the use of conventional semen decreased for artificial insemination in dairy heifers. • Sexed semen was selectively used for genetically superior, fertile heifers. • The conception rates after artificial insemination with sexed semen significantly decreased in early, warmer months. [ABSTRACT FROM AUTHOR]

Buffaloes have tendency to show seasonal reproduction and remain in anestrus due to limited ovarian activity during summer. The seasonal reproductive behavior is ascribed the effect of melatonin related to photoperiod. Treating animals with melatonin could be a possible strategy to overcome the problem. The role of MTNR1A gene has not been fully explained in the buffalo. Therefore, we conducted a study on 114 buffalo heifers to detect the polymorphic site in MTNR1A gene and further treated them with melatonin implants to investigate the role of most frequent genotype following melatonin treatment on pregnancy. The present investigation is the first to investigate the association between melatonin treated different MTNR1A genotype buffalo and pregnancy. We confirmed SNP at position 72 in 812 bp fragment exon II of MTNR1A gene. RFLP of PCR products with Hpa I enzyme resulted in three genotypes: TT (812bp), CT (812, 743, 69bp) and CC (743, 69bp). Next, buffaloes of each genotype (TT, CC, CT; n = 28 for each) were treated with melatonin implants to compare the conception rate with their corresponding untreated control (n = 10 for each genotype). Melatonin concentrations were higher (P < 0.05) for the treatment groups of all genotypes compared to their respective untreated control from day 1–28. The pregnancy rate was significantly associated with the MTNR1A genotype. The conception rate was higher (P < 0.05) for TT genotype than for the other genotypes of buffaloes treated with melatonin. Furthermore, buffaloes of TT genotype treated with melatonin started exhibiting estrus activity soon from second week of melatonin treatment (14.1 ± 2.1; range: 10–17 days) and were found to be 7.8 times more likely to become pregnant compared to other genotypes following melatonin treatment. In conclusion, TT genotype of MTNR1A gene is more sensitive to melatonin treatment that favours pregnancy in buffaloes during summer. • Buffalo has MTNR1A gene polymorphism. • TT genotype is the most sensitive for melatonin treatment. • TT genotype Buffalo are more likely to become pregnant after melatonin treatment. [ABSTRACT FROM AUTHOR]

Abstract Little is known of the genetic variability in semen quality traits in cattle and their inter-relationships. The objective of the present study was to estimate genetic parameters for a range of semen quality measures. The data consisted of 35,573 ejaculates from 787 artificial insemination bulls of 16 breeds. Genetic parameters were estimated using a repeatability animal linear mixed model. Large breed differences were detected with Belgian Blue bulls, on average, producing lesser semen volume than all other breeds while the Charolais bulls, on average, produced semen with fewer live sperm and reduced motility. The within-breed coefficient of genetic variation for sperm concentration, semen volume and total number of sperm per ejaculate was 0.17, 0.15 and 0.19, respectively. The genetic standard deviation for percentage live sperm pre-cryopreservation was 5.6% units while the genetic standard deviation for progressive motility pre-cryopreservation (scale 0 to 5) was 0.25 units. The heritability of all traits was between 0.13 and 0.34. The repeatability of the semen quality traits varied from 0.22 to 0.45. Sperm concentration and volume were negatively genetically correlated (−0.40) although the phenotypic correlation was near zero (−0.01). The genetic correlations between percentage live sperm and sperm motility varied from 0.68 to 0.94 irrespective of whether the traits were measured pre- or post-cryopreservation or even the change in both traits during cryopreservation. A very strong genetic correlation existed between percentage live sperm pre- and post-cryopreservation (0.96) or sperm motility pre- or post-cryopreservation (0.92). Results highlight the large genetic variability in a range of semen quality traits, many of which are actually highly heritable, and therefore useful predictors of actual phenotypic measures. Highlights • Considerable exploitable genetic variation exists in a range of semen quality traits. • Many of the semen quality traits are moderately heritable. • Breed differences in semen quality traits exist. • Antagonistic genetic correlations exist among some quality traits necessitating concurrent selection. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effect of a PGF 2 α-analogue (PGF) on ovulation and pregnancy rates after timed artificial insemination (TAI) in cattle. In Experiment 1 cows received an intravaginal progesterone-releasing device (CIDR) plus 2 mg im of estradiol benzoate (EB) on Day 0. The CIDR devices were removed on Day 8, and all cows received 150 μg im of d-cloprostenol (PGF 2 α-analogue), 300 IU of eCG and 1 mg of estradiol cypionate (ECP) im. On Day 9, cows were randomly assigned into two groups: 1) ECP Group (n = 17), that did not receive any further treatment; and 2) ECP-PG Group (n = 14) that were given 150 μg of d-cloprostenol (PGF) as adjuvant stimulus for ovulation. No difference between groups was detected in interval for ovulation (P = 0.5), and in the proportion of cows ovulating (P = 0.09). In Experiment 2, multiparous suckling crossbred Aberdeen Angus cows (n = 260), were treated into two groups, similarly as Experiment 1; ECP group (n = 122), and ECP-PG group (n = 138). All females were TAI on Day 10. The proportion of cows treated with ECP that became pregnant was 54.9% (67/122), and cows treated with ECP plus PGF was 55.1% (76/138; P = 0.9). In Experiment 3, 686 Nelore cows, 40 to 50 days postpartum, were treated as Experiment 1 (ECP group), however, on Day 8 cows were divided into 3 groups: ECP Group (n = 216); PGF-SC Group (n = 228), in which cows did not receive ECP and were given an additional subcutaneous injection of PGF on Day 8; and PGF–IM Group (n = 242), in which cows also did not receive ECP on Day 8 and were given an additional injection of PGF im on Day 9. On Day 10, estrus was evaluated at timed AI (TAI). There was no difference in the diameter of the dominant follicle at CIDR removal and at TAI, and pregnancy per AI among groups (P > 0.05). However, the proportion of cows that displayed estrus between CIDR removal and TAI was higher in ECP group than in PGF-SC and PGF-IM groups (P < 0.001). Cows that displayed estrus has higher P/AI than cows that did not (P = 0.008). In conclusion, these results suggested that intramuscular or subcutaneous injection of PGF 2 α could be successfully used to induce ovulation in cattle undergoing TAI, with similar pregnancy rates when compared with ECP. The subcutaneous injection of PGF on the same day of CIDR removal could be an interesting alternative due it reduces cattle management to obtain similar results. [ABSTRACT FROM AUTHOR]

Recent research has indicated that the interval between inseminations in modern dairy cattle is often longer than the commonly accepted cycle length of 18–24 days. This study analysed 257,396 inseminations in 75,745 cows from 312 herds in England and Wales. The interval between subsequent inseminations in the same cow in the same lactation (inter-service interval, ISI) were calculated and inseminations categorised as successful or unsuccessful depending on whether there was a corresponding calving event. Conception risk was calculated for each individual ISI between 16 and 28 days. A random effects logistic regression model was fitted to the data with pregnancy as the outcome variable and ISI (in days) included in the model as a categorical variable. The modal ISI was 22 days and the peak conception risk was 44% for ISIs of 21 days rising from 27% at 16 days. The logistic regression model revealed significant associations of conception risk with ISI as well as 305 day milk yield, insemination number, parity and days in milk. Predicted conception risk was lower for ISIs of 16, 17 and 18 days and higher for ISIs of 20, 21 and 22 days compared to 25 day ISIs. A mixture model was specified to identify clusters in insemination frequency and conception risk for ISIs between 3 and 50 days. A “high conception risk, high insemination frequency” cluster was identified between 19 and 26 days which indicated that this time period was the true latent distribution for ISI with optimal reproductive outcome. These findings suggest that the period of increased numbers of inseminations around 22 days identified in existing work coincides with the period of increased probability of conception and therefore likely represents true return estrus events. [ABSTRACT FROM AUTHOR]

This study addresses the hypothesis that SIRT1 upregulation prior to cryopreservation helps in recuperation from cryoinjury and improves the embryo quality. Bovine blastocysts were produced in vitro , and at Day 6 or 7 after fertilization, the blastocysts were cultured with 0 or 1 μM resveratrol for 6 or 24 h. This short duration of resveratrol treatment did not affect the embryo development or the grade of the blastocysts. However, both the durations of resveratrol treatment (6 h or 24 h) significantly increased the expression levels of SIRT1. When embryos pre-treated with resveratrol (0 or 1 μM) were cryopreserved and subsequently thawed, the hatching rates following 48 h of incubation were significantly higher for the resveratrol-treated embryos than for vehicle-treated embryos. Moreover, the resveratrol pretreatment significantly increased the copy number of mitochondrial DNA in the embryos, irrespective of the treatment durations. The in vitro -produced embryos at 6 days of insemination and the in vivo -developed embryos, which were collected from the donor cows at 6.5 days of insemination, were treated with resveratrol for 6 or 24 h prior to cryopreservation, respectively. The resveratrol pretreatment (for 6 or 24 h) resulted in high conception rate after thawing and transfer to the recipients, in both the in vivo and in vitro -produced embryos. In conclusion, our results suggest that pretreatment of bovine embryos with resveratrol improves the quality of embryos after cryopreservation and thawing probably through mitochondrial synthesis. [ABSTRACT FROM AUTHOR]

This study evaluated effects of equine chorionic gonadotropin (eCG) on fertility of 679 crossbred ( Bos taurus x Bos indicus ) lactating grazing cows synchronized for fixed-time AI (FTAI). At a random day of the estrous cycle cows received an intravaginal progesterone (P4) implant, 2 mg estradiol benzoate (EB) and 100 μg gonadorelin (D0-AM). On D7-AM, cows received 0.5 mg sodium cloprostenol and were randomly assigned into two treatments: eCG (n = 340; 400 IU eCG on D7), or Control (n = 339; no eCG). On D8-PM, P4 implants were removed and cows received 0.5 mg sodium cloprostenol and 1 mg EB. Insemination was performed on D10-AM. Pregnancy was diagnosed 30 and 60 d after AI. Treatment with eCG tended to increase pregnancy per AI (P/AI) compared to Control at 30 (37.8 vs. 30.2%; P = 0.06) and 60 (31.9 vs. 25.1%; P = 0.08) d. Pregnancy loss and twinning did not differ between groups. Treatment with eCG increased (P < 0.05) P/AI at 30 (39.0 vs. 25.2%) and 60 (32.8 vs. 21.3%) d for cows inseminated at ≤ 70 d in milk (DIM) but had no effect in cows receiving AI after 70 DIM. Thus, eCG on D7 of a FTAI protocol increased fertility of crossbred dairy cows inseminated in the early postpartum period. [ABSTRACT FROM AUTHOR]

The fertility of dairy cows has decreased dramatically worldwide over the last few decades, and several causes of this trend have been reported. Several studies have associated compromised udder health with deteriorating reproduction performance. Subclinical ketosis (SCK) has also been reported to be a risk factor for decreased conception. The objective of the present study was to describe how SCK might interact with the reported association between udder health and conception in dairy cows. Data from the French Milk Control Program and data on 8,549,667 instances of artificial insemination (AI) and their corresponding preceding and subsequent test-days from 5,979,701 Holstein cows were examined over a 5-year period (2008–2012). The effect of udder health was evaluated through a low (L) or high (H) somatic cell count (SCC) before and after AI using a threshold of 200,000 cells/mL, and transformed into four groups (LL, LH, HL, and HH). Three proxies for defining SCK were proposed based on the milk fat and protein content (or their ratio) before AI. Statistical analysis first included a generalized additive model to help define the optimal threshold values. Next, a logistic regression with a Poisson correction was performed. On average, the risk of conception at first AI was reduced by 14% for LH or HH cows (relative risk [and 95% CI] = 0.86 [0.85–0.87]) when the SCC increased or remained high within 40 days before and after AI, relative to LL group. The reduction of conception success associated with SCK (fat and protein contents changes) varied from 3% to 17% depending on the used SCK proxy. Including the interaction term SCC ∗ SCK clearly showed that the association of increased SCC around AI with conception success was modified by the presence of SCK. A cow that already has SCK and experiences an increase in SCC around or after AI exhibits up to 2 times further decrease in conception success compared with a cow with a high SCC and no SCK. In conclusion, this study reinforces the previously described association between intramammary inflammation around or after AI and a decreased rate of conception. These findings highlight how SCK interacts with the above-mentioned relationship by strengthening the negative association between mastitis and conception success. In addition, the present work supports the theory that local inflammation may affect the whole-body response and alter the functions of other organs, such as the reproductive tract. [ABSTRACT FROM AUTHOR]

The effects of eCG on follicular growth, ovulation, and pregnancy per artificial insemination (P/AI) in multiparous and primiparous Bos indicus beef cows submitted to timed artificial insemination (TAI) were evaluated in three experiments. In experiments 1 (follicular responses; n = 64), 2 (follicular growth and ovulation rate; n = 662), and 3 (P/AI; n = 2092), cows submitted to TAI were assigned to receive one of two treatments on Day 8 of the synchronization protocol: control (no additional treatment) or eCG (300-IU of eCG intramuscularly). In experiment 1, largest follicle (LF) diameter on Day 8 (P = 0.56) and the interval from progesterone (P4)-device removal to ovulation (P = 0.79) did not differ between treatments. However, the maximum diameter of the LF (P = 0.05) and ovulation rate (P = 0.03) were greater in cows that received eCG. In experiment 2, the diameter of the LF on Day 10, follicular growth, and ovulation rate were greater in eCG-treated cows (P < 0.01). However, CL diameter was similar between treatments (P = 0.11). In experiment 3, there was a treatment-by-parity interaction (P = 0.003) on P/AI, such that treatment with eCG was more effective in primiparous cows. In conclusion, eCG treatment resulted in increased final follicular growth, ovulation rate, and fertility in B indicus cows submitted to TAI protocols, especially in primiparous cows. [ABSTRACT FROM AUTHOR]

The objective of this study was to elucidate the impact of parity on the efficiency of three different protocols to synchronize time of ovulation in Holstein cows. All cows enrolled in this trial were categorized into primiparous and multiparous (M 1 = 2–3 and M 2 ≥3 parities). Conception (P/AI at Day 28) and pregnancy (P/AI at Day 75) rates in the Presynch and CIDRsynch (31.7% and 35.5%; 26.3% and 28.4%, respectively) groups were significantly greater than that in spontaneous estrus (SE; 24.7 and 20.4%, respectively) group (P = 0.048 and 0.024; 0.041 and 0.011, respectively); however, no significant differences were detected between the SE and Ovsynch group for conception, pregnancy, and embryonic loss rates. Conception and pregnancy rates in the Presynch group decreased from 37.8% and 33.5%, respectively in primiparous cows to 29.6% and 23.1%, respectively in M 1 cows (P = 0.022 and 0.007, respectively). However, conception and pregnancy rates using the CIDRsynch regimen were increased from 31.1% and 23.8% in primiparous cows to 41.4% and 34.7% in M 1 cows (P = 0.017 and 0.008, respectively), and 40.9% and 33.6% in M 2 cows (P = 0.021 and 0.019, respectively). Embryonic loss rate using the CIDRsynch protocol was decreased from 22.9% in primiparous cows to 15.1% and 18.2% in M 1 and M 2 cows (P = 0.013 and 0.130, respectively). On the contrary, embryonic loss rate using the Ovsynch protocol was increased from 11.5% in primiparous cows to 22.1% and 21.8% in M 1 and M 2 cows (P = 0.001 and 0.003, respectively). The Cox proportional-hazards model of embryonic loss showed significant associations for parity and season of calving with the hazard of embryonic loss (P = 0.001 and 0.016, respectively). Multiparous cows (M1 and M2) had a higher risk of embryonic loss than primiparous cows (Hazard ratio = 1.32 and 1.89, respectively). Our results indicate that use of the CIDRsynch regimen may achieve satisfactory conception and pregnancy rates in multiparous Holstein cows. However, synchronizing time of ovulation in primiparous cows with use of the Presynch treatment increases the fertility indices. [ABSTRACT FROM AUTHOR]

Heat stress has consequences on both the physiology and reproductive performance of cows, but the most dramatic effect for dairy producers is the decrease produced in fertility. The effects of heat stress on fertility include an increased number of days open, reduced conception rate, and larger number of cows suffering different types of anestrus. Once becomes pregnant, heat stress affects also the reproductive success of the cow through its direct effects on the ovary, uterus, gametes, embryo, and early fetus. This article reviews current knowledge of the effects of heat stress on fertility in dairy cows and the hormonal strategies used to mitigate these effects at the farm level. Administration of GnRH at the moment of artificial insemination can improve the conception rate. Breeding synchronization protocols for fixed-time insemination may reduce the calving conception interval and the number of services per conception. Progesterone-based protocols seem resolve better the reproductive disorders related to a hot environment (anestrus) than GnRH-based protocols. The use of combinations of GnRH, eCG, and hCG in progesterone-based protocols can improve results. Progesterone supplementation during the late embryonic and/or early fetal period would be useful in curtailing pregnancy losses, mainly in single pregnancies, whereas a more positive effect of treatment with GnRH than progesterone has been found in twin pregnancies. Melatonin therapy is emerging as a promising strategy to improve the natural reproductive performance of cows suffering conditions of heat stress. [ABSTRACT FROM AUTHOR]

Reproductive tract bacterial infections, particularly those caused by Escherichia coli and Trueperella pyogenes , can have a negative impact on reproductive performance. It has been hypothesized that the presence of E coli early postpartum may increase the risk of isolation of T pyogenes later postpartum. The objective of the present study was to examine associations between intrauterine bacterial infections with E coli and T pyogenes and any bacterial growth (irrespective of bacterial species), purulent vaginal discharge (PVD), cytologic evidence of endometritis (an increased proportion of polymorphonuclear cells [PMNs]), and reproductive performance. Dairy cows (n = 272) from six herds were examined at Days 0 (median, 2 days in milk), 21 and 42 postpartum. From each cow two intrauterine samples were collected via triple-guarded cytobrush at Days 0 and 21. The first cytobrush was used for bacteriologic culture. Escherichia coli and T pyogenes were isolated by culture, and E coli isolates were assigned to one of four phylogenetic groups using a two-step triplex polymerase chain reaction. In addition, T pyogenes was confirmed by polymerase chain reaction. The second cytobrush was used to prepare a cytology slide. Nucleated cells (n = 200) were categorized as epithelial cells, PMNs, or macrophages. Cows were also assessed for body condition score, PVD score, the presence of a CL, and pregnancy. Statistical analysis was performed using multivariable models. There was no association between the presence of E coli at Day 0 and probability of isolation of T pyogenes 3 weeks later; however, E coli positive cows at Day 0 were more likely to be diagnosed with E coli at Day 21 (relative risk [RR] = 2.0, P < 0.01). Escherichia coli at Day 0 or T pyogenes at Day 21 increased the risk of PVD diagnosis 3 weeks later (RR = 1.9; P = 0.04 and RR = 3.0; P = 0.05, respectively). Cows with any bacterial growth at Day 21, irrespective of species, were less likely to conceive within 3 weeks after the start of the seasonal breeding program (RR = 0.8; P = 0.05). Interestingly, cows with 25% PMNs or greater at Day 0 had shorter time to pregnancy (hazard ratio = 1.32; P = 0.05). Intrauterine bacterial infection may impair reproductive performance but the presence of E coli was not associated with isolation of T pyogenes 3 weeks later. Increased endometrial flux of PMNs in cows early postpartum may be a physiological process and improve reproductive performance. [ABSTRACT FROM AUTHOR]

Prebreeding vaccination should provide fetal and abortive protection against bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BoHV-1) but not impede reproduction when administered to cattle before estrus synchronization and breeding. The objective was to assess reproductive performance when naive beef heifers were vaccinated with modified-live viral (MLV) vaccine 2 days after unsynchronized estrus, and then revaccinated with MLV vaccine at 10 or 31 days before synchronized natural breeding. Sixty beef heifers naive to BVDV and BoHV-1 were randomly assigned to one of four treatment groups. Groups A and B (n = 20 per group) were vaccinated with MLV vaccine containing BVDV and BoHV-1 at 2 days after initial detected estrus, and then revaccinated 30 days later, which corresponded to 10 days (group A) or 31 days (group B) before synchronized natural breeding. Groups C and D (n = 10 per group) served as controls and were vaccinated with an inactivated vaccine that did not contain BVDV or BoHV-1 at the same time points as groups A and B, respectively. Estrous behavior was assessed using radio frequency technology. Estrus synchronization was performed, with initiation occurring at revaccination (groups A and C) or 21 days after revaccination (groups B and D). After synchronization, heifers were submitted to a bull breeding pasture for 45 days. At the end of the breeding period, heifers were assessed for pregnancy using ultrasonography. Progesterone concentrations were evaluated at estrus and 10 days after unsynchronized and synchronized estrus, at initial pregnancy check, and at the end of the study. All pregnant heifers in groups A and B and five pregnant heifers in group C were euthanized between 44 and 62 days of gestation and ovarian and conceptus tissues were assayed for BVDV and BoHV-1. Vaccination with MLV vaccine did not result in significant negative reproductive impact based on the duration of interestrus intervals, proportion of heifers exhibiting estrus within 5 days after synchronization, serum progesterone concentrations, pregnancy rates, and pregnancies in the first 5 days of the breeding season. Bovine viral diarrhea virus and BoHV-1 were not detected in luteal tissue, ovarian tissue, or fetal tissues. Use of MLV vaccine did not impede reproduction, when revaccination was performed at 10 or 31 days before synchronized natural breeding. [ABSTRACT FROM AUTHOR]

The primary objective was to determine if low doses of PGF2α (dinoprost) given intramuscularly (im) concurrent with timed artificial insemination (TAI) would improve conception rates in dairy cattle. A secondary objective was to determine if body condition score (BCS) and parity would influence conception rates, either independently or in interaction with PGF2α treatment. In experiment I, 307 lactating Holstein cows were randomly assigned to receive either 5-mg PGF2α im (PGF2α treated, n = 154) or 0-mg PGF2α (control, n = 153) at TAI (Day 0). Blood samples were obtained on Days −10, −3, 0, and 7 to determine plasma progesterone (P4) concentrations. Pregnancy was confirmed 30 to 32 days after insemination by transrectal ultrasonography. In experiment II, 451 cows were randomly assigned to receive either 10-mg PGF2α im (PGF2α treated, n = 226) or 0-mg PGF2α (control, n = 225) at TAI, and pregnancy was confirmed 45 to 50 days after TAI by palpation per rectum. Pregnancy data were analyzed by CATMOD (SAS). In experiment I, PGF2α treatment, BCS, and parity did not affect conception rate (35.7% vs. 37.0% for PGF2α treated vs. control; P > 0.05). However, in experiment II, conception rates were higher in cows given 10-mg PGF2α than those in control cows (45.8% vs. 36.0%; P < 0.05), in cows with high BCS than in cows with low BCS (52.1% vs. 30.4%; P < 0.01), and in primiparous than in multiparous cows (47.6% vs. 34.4%; P < 0.01), but their interaction with PGF2α treatment did not affect conception rates. In summary, 5 mg of PGF2α given im concurrent with TAI failed to enhance conception rate in lactating dairy cows, whereas 10 mg of PGF2α significantly increased conception rate. [ABSTRACT FROM AUTHOR]

Progesterone treatments are used to increase submission rates in postpartum dairy cows; however, in many cases the protocol is used as a blanket therapy for all cows without regard for physiological or disease state. The objective of this study was to identify the physiological or disease classes of cows that respond well (or not) to synchronization of estrus via progesterone. Dairy cows (n = 402) were monitored peri and postpartum to establish their physiological or disease status. Animals were classified as having negative energy balance, clinical lameness, uterine infection (UI), anovulatory anestrus, high somatic cell counts, and healthy (H). Blood samples were collected at five different time points and analyzed for metabolites. All animals received an 8-day controlled internal drug release protocol, which included GnRH at insertion and PGF 2α the day before removal. Response to the protocol was determined by visual observation of estrus synchronization. Conception rate was determined by ultrasonography between Days 32 and 35 after artificial insemination. Animals without UI were 1.9 times more likely to respond and two times more likely to be confirmed pregnant than those with UI. There was no relationship between negative energy balance and clinical lameness in the visual estrous response, but both conditions were associated with reduced conception rates. Dairy cows in anovulatory anestrus responded successfully to the protocol in both estrous response and conception rates. High glutathione peroxidase concentrations had a positive effect on conception rates, whereas high non-esterified fatty acids and beta-hydroxybutyrate had a negative effect on the estrous response. In conclusion, disease and physiological states of dairy cows determined the response to progesterone-based synchronization. The more disease or physiological problems the cows had, the lower the estrous response and conception rates; cows with these problems were not ideal candidates for synchronization. Both anestrus and healthy dairy cows were good responders to progesterone-based synchronization. [ABSTRACT FROM AUTHOR]

The influence of body condition score (BCS), rump fat thickness (RFAT), and live weight (LW), and the changes in these parameters during the interval from 165 of prepartum (i.e., 125 days of prior gestation) to 112 postpartum on first service conception and pregnancy rates were investigated in suckled Zebu ( Bos indicus ) beef cows (n = 266) subjected to timed artificial insemination (TAI) followed by natural mating. The aforementioned parameters were recorded at 165 ± 14 days (mean ± standard error) prepartum (concurrent with the weaning of previous calf), at parturition, and at 42 ± 7 days (at the onset of the synchronization of ovulation protocol), 82 ± 7 days (30 days after TAI), and 112 ± 7 days (60 days after TAI) postpartum. At the start of the breeding season (BS), cows were subjected to a synchronization of ovulation program for TAI. Bulls were placed with cows 10 days after TAI and remained until the end of the study (112 days postpartum). Cows with the highest BCS at parturition had an increased probability of first service conception rate at 60 days after TAI (P = 0.02) and a reduced probability of occurrence of pregnancy loss (P = 0.05). Also, cows had a greater likelihood of conceiving postpartum if they had greater RFAT and BCS at 165 ± 14 days prepartum (P = 0.01 and P = 0.03, respectively) and at parturition (P = 0.0007 and P = 0.003, respectively). Cows that had an increase in RFAT and BCS during the dry period (i.e., interval from weaning of the previous calf to parturition) also had a greater likelihood of conceiving (P = 0.03 and P = 0.06, respectively) during the BS. Among the different time points, RFAT and BCS at parturition had the largest impact on risk of conception during the BS. The LW was a poor predictor of conception during the BS (P = 0.11–0.68) except for LW at 165 ± 14 days prepartum (P = 0.01). Collectively, the findings indicated that the likelihood of conception during the BS was highest in cows that had an improvement in RFAT and BCS during the dry period. Therefore, assuring a good nutritional status in the dry period (BCS ≥ 3.0 at 165 ± 14 days prepartum and ≥3.25 at parturition) is an important aim to optimize the postpartum conception rate of suckled Zebu beef cows subjected to TAI followed by natural mating. [ABSTRACT FROM AUTHOR]

Although the snow leopard (Uncia uncia) is a common endangered felid species in zoos, little is known about the complex endocrine interactions controlling ovarian function and conception in this species. The goal of this work was to characterize ovarian activity throughout the estrous cycle, nonpregnant luteal phase (pseudopregnancy), and gestation in female snow leopards. This goal was accomplished using an enzyme immunoassay to measure fecal concentrations of estrogen metabolites (E) and progesterone metabolites (P). Fecal samples were collected from 12 female snow leopards (ages 18 months to 18 years) during one to three breeding seasons. In each breeding season, the majority of females (78%, 88%, and 100%, respectively) began to exhibit ovarian activity in December or January. The estrous cycle, defined by the first day of estrus (E ≥ 2 × basal concentration) to the first day of the subsequent estrus, was 12.7 ± 0.6 days (n = 145 cycles). Estrus lasted 4.3 ± 0.4 days with mean concentrations of fecal E during the follicular phase (1661 ± 139 ng/g feces) increasing 3.2-fold above basal concentrations (515 ± 32 ng/g feces). No spontaneous ovulations were observed in any of the cycling females. Nonpregnant luteal phases were observed in eight females that bred but did not become pregnant. The length of the nonpregnant luteal phase ranged from 11 to 72 days (45.7 ± 5.7 days; n = 10) with mean concentrations of fecal P during the luteal phase (12.46 ± 1.7 μg/g feces) increasing 6.2-fold above basal concentrations of P (2.01 ± 0.2 μg/g feces). Three of the females in the study became pregnant and gave birth after a gestation of 93 (n = 2) and 95 (n = 1) days. Fecal P concentrations during pregnancy increased to 11.64 ± 1.3 μg/g feces, or 5.8-fold above basal concentrations. The results of this study provide a comprehensive characterization of reproductive endocrinology in snow leopards, and confirm that fecal hormone monitoring is an effective way to monitor female snow leopards throughout the breeding season. [ABSTRACT FROM AUTHOR]

Abstract: The objectives of this retrospective study were to investigate the relationship between temperature–humidity index (THI) and conception rate (CR) of lactating dairy cows, to estimate a threshold for this relationship, and to identify periods of exposure to heat stress relative to breeding in an area of moderate climate. In addition, we compared three different heat load indices related to CR: mean THI, maximum THI, and number of hours above the mean THI threshold. The THI threshold for the influence of heat stress on CR was 73. It was statistically chosen based on the observed relationship between the mean THI at the day of breeding and the resulting CR. Negative effects of heat stress, however, were already apparent at lower levels of THI, and 1 hour of mean THI of 73 or more decreased the CR significantly. The CR of lactating dairy cows was negatively affected by heat stress both before and after the day of breeding. The greatest negative impact of heat stress on CR was observed 21 to 1 day before breeding. When the mean THI was 73 or more in this period, CR decreased from 31% to 12%. Compared with the average maximum THI and the total number of hours above a threshold of more than or 9 hours, the mean THI was the most sensitive heat load index relating to CR. These results indicate that the CR of dairy cows raised in the moderate climates is highly affected by heat stress. [Copyright &y& Elsevier]

Abstract: The curvilinear velocity (VCL) of boar spermatozoa between standard microscopy glassware decreases when the slides are coated with the hydrophobic polymer polystyrene (PS) compared with the less hydrophobic poly(methyl methacrylate) (PMMA) coating. Sperm from three boars were observed and analyzed using particle tracking software. The VCL did not differ significantly between coatings of different thickness, indicating no penetration of the sperm into the coating and that only the surface layer of the polymer film interacts with the sperm and buffer medium. The VCL of sperm between PS-coated surfaces was significantly reduced compared with PMMA surfaces (P < 0.0001), and this was attributed to a stronger hydrophobic effect between PS and water. The size of this effect varied between different boars, perhaps as a consequence of variations in hydrophobicity of sperm from different boars or different ejaculates. The modification of surface properties in this way may improve our understanding of sperm behavior and may provide improvements to assisted conception techniques as animal or human sperm used in assisted conception are frequently manipulated in laboratory plastics as part of diagnostic procedures (e.g., semen analysis) or before injection into an oocyte or during the co-incubation with the oocyte in IVF. Controlling the velocity of sperm using the interaction properties of inert polymer coatings could lead to new sperm selection procedures for clinical use or the development of model systems to better understand sperm–surface interactions. [Copyright &y& Elsevier]

Abstract: The ATP-binding cassette subfamily B member 1 (ABCB1) is an efflux transporter that excretes xenobiotics and waste matter. High expression of ABCB1 induced by forskolin (FSK) and rifampicin (RIF) in the bovine blastocysts reportedly improves the cellular quality. In the present study, interferon-α, similar to FSK and RIF, was highly potent in inducing the expression of ABCB1 in the bovine blastocysts but did not exhibit an additive effect with FSK and RIF. Bovine blastocysts stimulated by the combined treatment with FSK, RIF, and interferon-α to express high levels of ABCB1 displayed better freezing resistance as indicated by higher cell numbers in post thawing cultures. On transfer to recipients, such embryos established pregnancies with significantly higher frequencies in repeat breeder cows rather than normal ones. [Copyright &y& Elsevier]

Abstract: The aim of this study was to compare four methods of estrus resynchronization performed 23 days after timed artificial insemination (TAI) plus estrus observation in Bos indicus cows. Eight hundred fourteen lactating Nelore cows were submitted to TAI and then randomly assigned to one of the five following treatments: R23 (resynchronization without eCG), R23/200 (resynchronization with 200 IU of eCG), R23/300 (resynchronization with 300 IU of eCG), R23/TCR (resynchronization with temporary calf removal [TCR]), and a control group, with estrus observation followed by AI (with no resynchronization). Treatment consisted of a progesterone device plus administration of estradiol benzoate on Day 0; on Day 8, the device was removed and cloprostenol was applied, together with estradiol cypionate. Also on Day 8, either eCG was administered or TCR was performed in the resynchronized groups, except for R23. The females were inseminated 48 hours after device removal or TCR (33 days after the first TAI). The control group was kept under estrus observation from 18 to 23 days after the first TAI and was inseminated 12 hours after detection of estrus. The first pregnancy evaluation was performed using ultrasound examination 31 days after the first TAI. After 30 days of the resynchronization, a second pregnancy evaluation was performed and the animals in the R23/300 and R23/TCR groups achieved the highest conception rates, 76.6% and 74.0%, respectively (P < 0.05). There were no differences between the conception rates of the animals in the R23/200 (63.3%), R23 (61.3%), and control (54.3%) groups (P > 0.05). These results suggest that estrus resynchronization at 23 days after TAI can effectively improve the conception rate of lactating Bos indicus cows in a short time period. Furthermore, resynchronization with 300 IU of eCG or with TCR provided the best results. [Copyright &y& Elsevier]

Abstract: Crossbred beef heifers (N = 59) were vaccinated at the time of synchronization/breeding with either a commercially available bovine herpesvirus type 1 modified live virus (MLV) (one dose) or inactivated virus vaccine (one or two doses). The estrus cycle was synchronized at vaccination and heifers were artificially inseminated 8 days (one dose) or 36 days (two dose) after initial vaccination. Pregnancy rates were greater for control heifers (90%; P = 0.02) and heifers given the inactivated virus vaccine (one dose: 86%; P = 0.08; or two: 90%; P < 0.01) than those given the MLV vaccine (48%). No control heifers experienced an abnormal estrous cycle, whereas only two (two dose; 2/21) and one (one dose; 1/7) heifers in the inactive virus groups had abnormal estrous cycles and were similar to control (P > 0.10). Heifers given the MLV vaccine had a greater (P = 0.02) percentage of abnormal estrous cycles (38%; 8/21) compared with the control and inactivated groups. Of the heifers with an abnormal estrous cycle, 100% of heifers given the inactivated vaccine (one or two dose) conceived at their return estrus, whereas only 38% of heifers given the MLV vaccine conceived at their return estrus (P > 0.10). During the synchronization period, concentrations of estrogen were greater (P < 0.01) in the control and the two-dose inactivated group compared with the MLV group. After AI, progesterone concentrations were greater (P < 0.01) in control heifers compared with the inactivated and MLV groups, but were similar (P ≥ 0.18) between the inactivated and MLV groups. Therefore, naïve heifers vaccinated with the inactivated vaccine were less likely to have an abnormal estrous cycle and had significantly higher pregnancy rates compared with heifers vaccinated with the MLV vaccine. In summary, vaccination of naïve heifers with an MLV vaccine at the start of a fixed-time AI protocol had a negative effect on pregnancy success. [Copyright &y& Elsevier]

Abstract: An adjustment of sex ratio of offspring to the conditions present at conception is seen in many mammals including horses. This depends on preferential survival of male embryos under conditions of high energy intake. In several species, growth factors including insulin like growth factor (IGF)-1 have been shown to promote embryonic development by decreasing apoptosis and increasing cell proliferation. We hypothesized that sex-related differences in IGF-1 expression in equine embryos during the phase of maternal recognition of pregnancy might exist and thus contribute to preferential survival of embryos from either of both sexes under specific environmental conditions. Insulin like growth factor-1 mRNA expression of in vivo-produced equine embryos on different days of pregnancy (Day 8, N = 6; Day 10, N = 8; Day 12, N = 14) was analyzed. Insulin like growth factor-1 mRNA expression was evaluated by reverse transcription quantitative polymerase chain reaction. The sex of the embryo was determined by detection of X-inactivation specific transcript (Xist) RNA and equine sex determining region of the Y chromosome DNA. Embryos positive for Xist expression were classified as female, and Xist negative and equine sex determining region of the Y chromosome positive embryos were classified as male. From 28 embryos tested, 15 (54%) showed positive Xist expression and were thus classified as female. Insulin like growth factor-1 mRNA expression was influenced by sex (P = 0.01) but not by day of pregnancy (relative expression of IGF-1 in relation to β-actin, Day 8: male 5.1 ± 2.1, female 11.4; Day 10: male 5.2 ± 1.6, female 17.4 ± 6.7; Day 12: male 2.6 ± 0.3, female 11.6 ± 2.4). Results demonstrate an increased expression of IGF-1 in female equine embryos. Sex-related influences on expression of the IGF system are probably related to a gradual X chromosome inactivation. [Copyright &y& Elsevier]

Abstract: Cryopreservation of bovine embryos can be performed by a variety of methods with variable degree of success. Here, we report a new, easy to perform, simple, inexpensive, and successful method for vitrification of bovine blastocysts. In vitro produced bovine blastocysts were exposed to vitrification solution (5.5 m ethylene glycol, 10% serum and 1% sucrose) in one single step for 20 s, loaded on a paper container prepared from commonly available non-slippery, absorbent writing paper, and then were directly plunged into liquid nitrogen for storage. Vitrified blastocysts were warmed by serial rinsing in 0.5, 0.25 and 0.125 m sucrose solution for 1 min each. Results showed that one step exposure of bovine blastocysts to cryoprotective agents was sufficient to achieve successful cryopreservation. Under these conditions, more than 95% of blastocysts survived the vitrification-warming on paper containers which was significantly higher than those obtained from other containers, such as electron microscope (EM) grid (78.1%), open pulled straw (OPS; 80.2%), cryoloop (76.2%) or plastic straw (73.9%). Embryo transfer of blastocysts vitrified-warmed on paper container resulted in successful conception (19.3%) and full-term live birth of offspring (12.3%) which were lower (P < 0.05) than those obtained from non-vitrified blastocysts (38.0 and 32.7%) but were comparable (P > 0.05) to those obtained from blastocysts vitrified-warmed on EM grid (23.3 and 14.2%). Our results, therefore, suggest that paper may be an inexpensive and useful container for the cryopreservation of animal embryos. [Copyright &y& Elsevier]

Abstract: Using two PGF2α treatments 14 days apart as a way to enhance estrus detection rate following the 2nd treatment is a reproductive management tool that continues to be used on large dairy farms. In one study, in cows with a functional CL and a dominant follicle, treatment with cloprostenol vs. dinoprost resulted in greater peripheral estradiol concentrations. The objective of the present study was to determine if cloprostenol could enhance pregnancy rates of cows in a large dairy herd using a PGF2α program for 1st artificial insemination (AI). Lactating dairy cows (n = 4549) were randomly assigned to receive two treatments of either 500 μg cloprostenol or 25 mg dinoprost 14 days apart, with the 2nd treatment on the 1st day of the voluntary waiting period (57 DIM). Cows detected in estrus within 5 days after the 2nd treatment were inseminated. There was no effect of treatment on day of estrus detection, with 78% of cows inseminated on Days 3 or 4 following treatment. Cloprostenol increased (P < 0.01) estrus detection rates in 1st parity cows compared to dinoprost, 42.4 vs. 34.0%. In cows inseminated on Days 3 or 4 after treatment, cloprostenol increased (P = 0.05) conception rates compared to dinoprost, 38.3 vs. 34.4%. When treatments and parities were combined, conception rates increased (P < 0.02) with interval after treatment (27.0, 36.4, and 44.5% for Days 1 or 2, Days 3 or 4, and Day 5, respectively). Cloprostenol increased (P = 0.02) overall pregnancy rate compared to dinoprost, 14.4 vs. 12.2%. In summary, cloprostenol increased fertility in 1st parity cows inseminated on Days 3 or 4 following treatment and subsequently enhanced pregnancy rates of 1st parity lactating dairy cows compared to dinoprost. Fertility appeared greater in cows expected to have had a young antral ovarian follicle at treatment. [Copyright &y& Elsevier]

Abstract: The objective was to evaluate if uterine sampling by low-volume uterine lavage, done shortly before the end of the voluntary waiting period, affected reproductive performance, milk production, or culling. Approximately 20 cows (40 and 60 d postpartum) were sampled in each of 38 herds (total, 752 cows), whereas all remaining cows concurrently between 40 and 60 d were a non-sampled reference cohort (n = 2,252 cows). There was an interaction between parity and sampling for first-service conception, calving to conception interval, and milk production. When stratified by parity, there was a tendency for impaired reproductive performance in sampled primiparous cows compared with non-sampled primiparous cows (Odds Ratio for pregnancy to first-service = 0.78 (95% C.I. 0.58–1.04). The Hazard Ratio for pregnancy in sampled primiparous cows was 0.89 (95% C.I. 0.77–1.04) compared to non-sampled primiparous cows. Sampling did not affect first-service conception rate (Odds Ratio for pregnancy Odds ratio (OR) for pregnancy = 1.03; 95% C.I. 0.80–1.33) or calving-to conception interval in multiparous cows (Hazard Ratio = 1.04; 95% C.I. 0.91–1.18). Sampling did not affect culling risk (Hazard Ratio HR = 0.92; 95% C.I. 0.77–1.11) after accounting for covariates. After stratification by parity, milk production was not affected by sampling except in fourth- and fifth-parity cows where sampled cows produced more milk than non-sampled cows after controlling for first test-day milk production and days-postpartum at first test-day. In conclusion, sampling by low-volume uterine lavage did not have significant detrimental effects on reproduction, culling, or milk production. However, there was a tendency for lower first-service conception in sampled primiparous cows, but the procedure appeared to be benign in multiparous cows. [Copyright &y& Elsevier]

Abstract: The objective was to improve the reproductive performance of a prostaglandin (PG) F2α-based protocol for timed artificial insemination (TAI) in sheep (Synchrovine®: two doses of 160 μg of delprostenate 7 d apart, with TAI 42 h after second dose). Three experiments were performed: Experiment 1) two doses of a PGF2α analogue (delprostenate 80 or 160 μg) given 7 d apart; Experiment 2) two PGF2α treatment intervals (7 or 8 d apart) and two times of TAI (42 or 48 h); and Experiment 3) insemination 12 h after estrus detection or TAI with concurrent GnRH. Experiments involved 1131 ewes that received cervical insemination with fresh semen during the breeding season (32/34 °S–58 °W). Estrous behaviour, conception rate, prolificacy, and fecundity (ultrasonography 30–40 d), were assessed. In Experiment 1, ewes showing estrus between 25 and 48 h or at 72 h after the second PGF2α did not differ between 80 and 160 μg of delprostenate (73 vs 86%, P = 0.07; and 92 vs 95%, P = NS, respectively). Conception rate and fecundity were lower (P < 0.05) using 80 vs 160 μg (0.24 vs 0.42, and 0.27 vs 0.47, respectively). In Experiment 2, giving PGF2α 7 d apart resulted in higher (P < 0.05) rates of conception (0.45 and 0.51) and fecundity (0.49 and 0.53) than treatments 8 d apart (conception: 0.33 and 0.29; fecundity: 0.33 and 0.34) for TAI at 42 and 48 h, respectively. In Experiment 3, rates of conception, prolificacy and fecundity were similar (NS) between Synchrovine® with TAI at 42 h (0.50, 1.13, and 0.56) and AI 12 h after estrus detection (0.47, 1.18, and 0.55), and Synchrovine® plus GnRH at TAI (0.38, 1.28, and 0.49). However, all TAI treatments had lower (P < 0.05) prolificacy and fecundity compared to AI following detection of spontaneous estrus (1.39 and 0.83, respectively). In conclusion, the Synchrovine® protocol was: a) more successful using 160 vs 80 μg delprostenate; b) more successful with a 7 d than 8 d PGF2α interval; c) similarly effective for TAI versus AI 12 h after estrus detection; and d) not improved by giving GnRH at TAI. [Copyright &y& Elsevier]

Abstract: The objective was to determine the association of mRNA expression of cystine rich secretary protein 2 (CRISP2), chaperonin containing T-complex protein 1, subunit 8 (CCT8), and phosphatidylethanolamine-binding protein 1 (PEBP1), in sperm of Holstein bulls with Sire Conception Rate (SCR) scores between −4 and +4. These proteins were involved in sperm capacitation and sperm-egg fusion. Samples of sperm obtained on a single day from Holstein bulls (N = 34) in a commercial AI centre were used to evaluate relative mRNA expression of CRISP2, CCT8, and PEBP1. The mRNA abundance of CRISP2 was positively correlated (r = 0.88; P < 0.002), CCT8 was negatively correlated (r = −0.87; P < 0.002), and PEBP1 was positively correlated (r = 0.83; P < 0.006) with SCR-scores. The means of CRISP2 mRNA abundance was greater among positive SCR-score bulls (2.5 to 8 fold), the means of CCT8 mRNA abundance was greater among the negative SCR-score bulls (9.5 to 3.5 fold), and the means of PEBP1 mRNA abundance was greater for the positive SCR-score bulls (5.4 to 7.7 fold). In multivariate regression models predicting SCR-scores, mRNA abundance of CCT8 was significantly associated with SCR-score in all models. In the presence of CRISP2 mRNA abundance in the model, the SCR score''s predictability of PEBP1 was insignificant. However, in the absence of CRISP2 mRNA abundance in the model, the SCR-score''s predictability of PEBP1 was significant. In multivariate regression models, CRISP2 and CCT8 mRNA expression in sperm accounted for 95% of the variance in Holstein bull''s SCR-scores. In conclusion, Holstein bulls with greater CRISP2 and lower CCT8 mRNA expression in sperm had higher probabilities of siring calves. [Copyright &y& Elsevier]

Abstract: Yearling Bos indicus × Bos taurus heifers (n = 410) from three locations, were synchronized with either the Select Synch/CIDR+timed-AI (SSC+TAI) or 7-11+timed-AI (7-11+TAI) treatments. On Day 0 of the experiment, within each location, heifers were equally distributed to treatments by reproductive tract score (RTS; Scale 1–5: 1 = immature, 5 = estrous cycling) and body condition score. The 7-11+TAI treatment consisted of melengestrol acetate (0.5 mg/head/d) from Days 0 to 7, with PGF2α (25 mg im) on Day 7, GnRH (100 μg im) on Day 11, and PGF2α (25 mg im) on Day 18. The SSC+TAI heifers received the same carrier supplement (without MGA) from Days 0 to 7, and on Day 11 they were given 100 μg GnRH and an intravaginal CIDR (containing 1.38 g progesterone). The CIDR were removed on Day 18, concurrent with 25 mg PGF2α im For both treatments, estrus was visually detected for 1 h twice daily (0700 and 1600 h) for 72 h after PGF2α, with AI done 6 to 12 h after a detected estrus. Non-responders were timed-AI and received GnRH (100 μg im) 72 to 76 h post PGF2α. The 7-11+TAI heifers had a greater (P < 0.05) estrous response (55.2 vs 41.9%), conception rate (47.0 vs 31.3%), and synchronized pregnancy rate (33.5 vs 24.8%) compared to SSC+TAI heifers, respectively. Heifers exhibiting estrus at 60 h (61.7%) had a greater (P < 0.05) conception rate compared to heifers that exhibited estrus at ≤ 36 (35.3%), 48 (31.6%), and 72 h (36.2%), which were similar (P > 0.05) to each other. As RTS increased from ≤ 2 to ≥ 3, estrous response, conception rate, synchronized pregnancy rate, and 30 d pregnancy rate all increased (P < 0.05), irrespective of synchronization treatment. In conclusion, the 7–11+TAI treatment yielded greater synchronized pregnancy rates compared to SSC+TAI treatment in yearling Bos indicus × Bos taurus heifers. [Copyright &y& Elsevier]

Abstract: A sample of 423 Sarda ewe lambs from three different farms was used to evaluate the effect of one or two melatonin implants on the time of first conception. On each farm, 141 animals were divided into three groups. On June 30 these animals received either no treatment (Group C), 18 mg melatonin (Group M1, one implant), or 18 + 18 mg melatonin (Group M2, two implants). Thirty-five days after treatment, rams were introduced in the ewe lambs flock and subsequently removed after 40 days. Lambing dates were recorded between 150 and 190 days from the first day of male introduction. Genotyping and sequencing of the MT1 exon 2 were carried out to analyze the structure and the possible influence of the MT1 receptor gene on reproductive response to melatonin treatment. Melatonin-treated animals had a higher rate of pregnancy (P < 0.05) and lambed earlier (P < 0.05) compared with untreated animals. Single nucleotide polymorphisms were found in exon II of MT1 gene at positions C606T and G612A leading to genotypes C/C, C/T or T/T and +/+, +/− and −/−, respectively. Melatonin-treated animals of +/+ genotype showed a higher number of pregnancies (P < 0.05) and lambed earlier (P < 0.05) compared to untreated animals of the same genotype. Melatonin treatment did not affect reproductive activity in any other genotype analyzed. No correlation between genotype and the time of first conception was found in untreated animals. Concluding data revealed the positive effect of melatonin treatment on the time of first conception in ewe lambs and highlighted that +/+ genotype is able to influence reproductive response to melatonin treatment. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to determine the prevalence and risk factors for subclinical endometritis (SE) and its effects on fertility in repeat breeder dairy cows. Dairy cows of parity 1 to 5 that were artificially inseminated (AI) 3 or more times (selected cows were artificially inseminated an average of 3.9 times) were examined at 190 ± 40 days in milk, and clinically normal cows (n = 77) were selected based on the absence of abnormal discharges on external inspection and the absence of abnormal findings on transrectal palpation and ultrasonographic examination. Endometrial samples were collected from the uterus by using the lavage technique in the luteal phase of the estrus cycle. Collected samples were centrifuged and a drop of sediment was streaked on to a clean microscopic slide and stained with Giemsa. The percentage of polymorphonuclear cells (neutrophils) was counted for each specimen. The analysed data showed that the average amount of neutrophils was 3.1% (0–9) in the selected cows. Abnormal calving (dystocia, twin births, and abortion), retained placenta, and postpartum uterine infections were associated with an increase in prevalence of SE. Subsequently, SE was significantly associated with a decrease in conception rate in the next AI. Conception rate in the next AI was 5% for cows (n = 38) with SE (≥ 3% neutrophil), and 47% for cows (n = 34) without SE (< 3% neutrophil) (P = 0.001). The prevalence of cytologically diagnosed SE (≥ 3% neutrophil) was 52.7% (n = 38). In conclusion, abnormal calving, retained placenta, and postpartum uterine infections may be associated with an increase in prevalence of SE and subsequently, SE may decrease reproductive performance and increase the incidence of repeat breeder syndrome. [Copyright &y& Elsevier]

Abstract: The net economic value of reproductive efficiency in dairy cattle was estimated using a stochastic dynamic simulation model. The objective was to compare the economic consequences of reproductive performance scenarios (“average” and “poor”) of a cow having a good reproductive performance and to explore which reproductive factors have an important impact on economic efficiency. A “good” reproductive performance scenario was defined with 1 ovulation rate (POVUi), 0.7 estrus detection rate (PEst), 0.7 conception rate (PCon), 0.03 incidence rate of postpartum disorders prolonging the ovarian cyclicity (CO), 0.2 incidence rate of postpartum disorders reducing conception (ME), 0.05 embryonic death rate (ED), and voluntary waiting period (VWP) of 9 wks pp (post partum). In the current situation of dairy cows in the Netherlands, an “average” reproductive scenario (0.95 POVUi, 0.5 PEst, 0.5 Pcon, 0.07 CO, 0.27 ME, 0.07 ED and VWP of 12 wks pp) and a “poor” reproductive scenario (0.90 POVUi, 0.3 PEst, 0.3 Pcon, 0.11 CO, 0.33 ME, 0.09 ED and VWP of 15 wks pp) were identified. A sensitivity analysis was performed by comparing changes of single effect of factors in a good and poor scenario with the average scenario. The mean net economic loss (NELi) compared with the good scenario was €34 and €231 per cow per year for the average and poor reproductive performance scenario, respectively. Increasing the calving interval resulted in greater economic loss. The important factors on the cost of reproductive efficiency were the involuntary culling cost and the return of milk production. Variation in PCon, PEst, ME, ED, and VWP had large impacts on economic benefits. [ABSTRACT FROM AUTHOR]

Abstract: The ECF™ lateral flow assay test is marketed to detect non-pregnancy in mares. The objectives of the present study were to determine the accuracy of the ECF test, the accuracy of the electronic reader accompanying the ECF test, and agreement between two human readers and the electronic reader. Serum samples were collected from anestrus, cycling but not inseminated, and inseminated mares, and were evaluated with the ECF™ test (EDP Biotech Company, Knoxville, TN, USA) at The Ohio State University and at the EDP Biotech Laboratory. Specificity ranged from 0.07 to 0.16, the negative predictive value ranged from 0.15 to 0.33, and accuracy ranged from 0.43 to 0.52. The electronic reader did not add improve the accuracy or predictive values of the test. Based on the electronic reader, 80.0% of the serum samples collected from the anestrus mares were false positives; Readers 1 and 2 had 60.0 and 33.3% false positives, respectively. For samples collected during the estrous cycle, 83.9% were false positives by the electronic reader, whereas Readers 1 and 2 had 43.7 and 26.4% false positives. We concluded that, regardless of whether the test strips were evaluated by a human or electronic reader, this assay was not accurate for determination of the non-pregnant mare. [Copyright &y& Elsevier]

Abstract: The objective of this study was to investigate the prevalence of subclinical endometritis 4h after AI and its effect on first service conception rate (FSCR) in dairy cows. A total of 201 Holstein-Friesian cows with no signs of clinical endometritis were examined 4h after first AI for signs of subclinical endometritis. Endometrial samples were collected from the uterus using the cytobrush technique. The proportion of polymorphonuclear cells (PMN) in the cytological sample was used to characterize an inflammation of the endometrium. Cows were categorized into three groups according to the proportion of PMN in the sample. Cows with 0% PMN (n =115) were assigned to group Zero, cows with >0–15% PMN (n =59) to group Medium, and cows with >15% PMN (n =27) to group High. Pregnancy diagnosis was performed between days 38–44 after AI by palpation of the uterus and its contents per rectum. The FSCR was significantly higher in group Medium than in groups Zero and High (57.6% vs. 39.1% and 29.6%). Statistical analysis revealed an interaction between parity and PMN group. Primiparous cows were at higher risk of being classified into group Medium than multiparous cows (OR=2.27, P =0.01). Primiparous cows in group Zero had lower odds of pregnancy after first AI than primiparous cows in group Medium (OR=0.3, P =0.02). A comparison with cows that were not examined for subclinical endometritis showed that the collection of endometrial samples itself had no effect on FSCR. [Copyright &y& Elsevier]

Abstract: The objective of this study was to compare conception rates, non-return rates and sex ratios of sexed and conventional semen from the same sires in commercial dairy herds in Denmark. The semen was produced from three bulls from each of the three major dairy breeds in Denmark: Holstein, Jersey and Danish Red Dairy Breed (nine bulls total), in order to answer questions on breeds differences in field results. AI was performed by trained technicians using a minimum of 150 doses of sorted sperm and 50 control doses from each bull. During the trial, a total of 2087 doses were used in 63 herds. The trial showed that the conception rate using sorted semen was 5% points lower than with conventional doses for Danish Reds, 7% points for Jerseys, and 12% points for Holsteins. Translating this into non-return rate revealed differences of 10–20% points among bulls. These differences are thought to be a good indicator of what to expect from commercial use of sexed semen. The sex ratios varied from 89% to 93% female calves among breeds, which on average is consistent with the theoretical average sex ratio of 93% females considering the low number of inseminations. [Copyright &y& Elsevier]

Abstract: This study was designed to examine estrous response rates to the therapeutic treatment of clinical anestrus in high producing dairy cows and to identify the factors that could affect these rates. Cows with silent ovulation (Subestrus group), cystic ovarian disease (Cyst group) or ovarian hypofunction (OH group) were given specific treatment for their disorder. Data were derived from 1764 treatments in cows producing a mean of 45.4kg of milk upon treatment including: 889 subestrous cows, 367 cystic cows and 508 cows with ovarian hypofunction. Cows showing estrus following treatment exhibited a similar pregnancy rate to cows attaining natural estrus used as reference: 33% (337/1006) and 35% (626/1796), respectively. No significant differences in pregnancy rates were observed among the Subestrus, Cyst and OH groups (34% (196/571), 34% (44/130), 32% (97/305), respectively. Based on the odds ratio, an estrous response for all groups was less likely to occur in cows that had suffered previous anestrus, compared to cows that were anestrous for the first time, whereas the likelihood of an estrous response increased in cows treated after 90 days in milk. Our results indicate that previous anestrus and a late stage of lactation can have a negative and positive effect, respectively, on the estrous response to the specific treatment of clinical anestrus shown by high producing dairy cows. Treatment targeted at each type of clinical anestrus can render similar pregnancy rates to those shown by cows in natural estrus. [Copyright &y& Elsevier]

Abstract: The present study was conducted to establish a simple and efficient method of producing monozygotic twin calves using the blastomere separation technique. To produce monozygotic twin embryos from zona-free two- and eight-cell embryos, blastomeres were separated mechanically by pipetting to form two demi-embryos; each single blastomere from the two-cell embryo and tetra-blastomeres from the eight-cell embryo were cultured in vitro using the Well of the Well culture system (WOW). This culture system supported the successful arrangement of blastomeres, resulting in their subsequent aggregation to form a demi-embryo developing to the blastocyst stage without a zona pellucida. There was no significant difference in the development to the blastocyst stage between blastomeres separated from eight-cell (72.0%) and two-cell (62.0%) embryos. The production rates of the monozygotic pair blastocysts and transferable paired blastocysts for demi-embryos obtained from eight-cell embryos (64.0 and 45.0%, respectively) were higher than those for demi-embryos obtained from two-cell embryos (49.0 and 31.0%, P <0.05). The separated demi-embryos obtained from eight-cell embryos produced by IVM/IVF of oocytes collected by ovum pick-up (OPU) from elite cows and cultured in wells tended to have a higher pregnancy rate (78.9% vs. 57.1%) and similar monozygotic twinning rate (40.0% vs. 33.3%) compared with monozygotic twin blastocysts obtained by the conventional bisection of in vivo derived blastocysts. In conclusion, producing twins by separation of blastomeres in OPU-IVF embryos, followed by the WOW culture system, yielded viable monozygotic demi-embryos, resulting in high rates of pregnancy and twinning rates after embryo transfer. [Copyright &y& Elsevier]

Abstract: The high incidence of pregnancy loss and prenatal morbidity and mortality in cloned animals may be due to placental insufficiency, thereby compromising fetal survival. Our objective was to characterize morphological changes in fetal membranes of cloned bovine pregnancies. Two groups of cows with cloned fetuses, produced by two cloning techniques, a commercial group (n =16) and a hand-made group (n =4), and control fetuses derived from traditional embryo transfer (n =6) or AI (n =6), were compared at various stages of gestation (Days 80, 120, 150, 180, 210, and 240; Day 0=estrus). Thickness and shape of the amniotic membrane, placentome shape and length, umbilical cord shape and diameter, and fetal fluid echodensities were assessed by ultrasonography, and the placenta was evaluated histologically. Only eight (40%) of cloned pregnancies reached term and seven calves (35%) were alive at birth. Both placentome length and umbilical cord diameter were larger (P <0.05) in clones than in normal fetuses at all stages of gestation. Amniotic membrane abnormalities (Day 120) including focal edema and the presence of a series of nodules were detected in 38% of the clones and were always accompanied by hyper-echodense spikes or irregularities (detected ultrasonographically) around the umbilical cord. Histopathology revealed degenerate inflammatory cells, edematous chorioallantoic membranes, and decreased epithelial thickness. We inferred that these morphological anomalies of placentomes compromised fetal development, and we concluded that ultrasonographic monitoring of pregnancies enabled characterization of changes in the placentae and may be useful to assess fetal well-being. [Copyright &y& Elsevier]

Abstract: Repeat breeding is an important factor affecting economic success in dairy management. The objective of this study was to investigate the effectiveness of transfer of frozen-thawed IVF embryos in establishing pregnancy in repeat-breeding Holstein cattle. Cumulus oocyte complexes were collected by aspiration of 2–5mm follicles from ovaries obtained at two local abattoirs. After IVF, days 7 and 8 blastocysts were frozen either in 1.5M ethylene glycol with 0.1M sucrose, or in 1.4M glycerol with 0.1M sucrose. Holstein recipients (122 heifers and 410 cows) included those that had not conceived after 3–21 inseminations. Embryos frozen in ethylene glycol were transferred directly, and embryos frozen in glycerol were transferred after dilution of the cryoprotectant in sucrose into recipients 7 or 8 days after estrus (without-AI group), or following AI (with-AI group). Pregnancy rates were compared by the Chi-square test. Significantly higher pregnancy rates were achieved by embryo transfer following AI (with-AI group) than by embryo transfer alone (without-AI group) in both heifers (49.2 and 29.5%, respectively) and cows (41.5 and 20.4%, respectively; P <0.05). Pregnancy rates were not significantly different between heifers and cows. However, pregnancy rate decreased as the number of inseminations prior to embryo transfer increased in the with-AI group, but not in the without-AI group. Therefore, transfer of frozen-thawed IVF embryos during the same cycle in which AI was done improved pregnancy rates in repeat-breeding Holstein heifers and cows, and suggested that embryo transfer is an alternative in the treatment of repeat breeding. [Copyright &y& Elsevier]

Abstract: The aims of the present study were to evaluate and compare the accuracy of transrectal ultrasonography and pregnancy-associated glycoprotein radioimmunoassay (PAG-RIA) test for diagnosis of pregnancy in buffaloes. Two hundred and seventy-five buffalo cows and heifers were examined once for pregnancy diagnosis by transrectal ultrasonography using a 5MHz linear-array transducer between Days 19 and 55 after mating. After ultrasound scanning, a blood sample was withdrawn from jugular vein of each animal for measuring pregnancy-associated glycoprotein using a heterologous double-antibody RIA. Based on palpation of the uterus per rectum at Days 75–90, 87 animals were designated pregnant and 188 as non-pregnant. The sensitivity of transrectal ultrasonography at Days 19–24 was 44.4%, reaching 100% from Day 31 after mating. The specificity of transrectal ultrasonography ranged between 92.5 and 100% from Days 19 to 55 after mating. The sensitivity of PAG-RIA test was 11.1% at Days 19–24 and reached 100% from Day 31 after mating. The specificity of PAG-RIA test ranged from 90 to 100% from Days 19 to 55 after mating. There were no significant differences between the sensitivity and specificity of the two tests in all examined periods. In conclusion, transrectal ultrasonography and PAG-RIA test are highly accurate tests for detecting pregnant buffaloes from Day 31 after mating onwards. [Copyright &y& Elsevier]

Abstract: This study was conducted to examine pregnancy progression and fetal characteristics following transfer of vitrified bovine nuclear transfer versus in vivo-derived embryos. Nuclear transfer (NT) was conducted using cumulus cells collected from an elite Holstein–Friesian dairy cow. Expanding and hatching blastocysts on Day 7 were vitrified using liquid nitrogen surface vitrification. Day 7 in vivo embryos, produced using standard superovulation procedures applied to Holstein–Friesian heifers (n =6), were vitrified in the same way. Following warming, embryos were transferred to synchronized recipients (NT: n =65 recipients; Vivo: n =20 recipients). Pregnancies were monitored by ultrasound scanning on Days 25, 45 and 75 and a sample of animals were slaughtered at each time point to recover the fetus/placenta for further analyses. Significantly more animals remained pregnant after transfer of in vivo-derived embryos than NT embryos at all time points: Day 25 (95.0 versus 67.7%, P <0.05), Day 45 (92.8 versus 49.1%, P <0.01) and Day 75 (70.0 versus 20.8%, P <0.0). There was no significant difference (P =0.10) in the weight of the conceptus on Day 25 from NT transfers (1.14±0.23g, n =8) versus in vivo transfers (0.75±0.19g, n =8). On Day 45, there was no significant difference in the weight of either fetus (P =0.393) or membranes (P =0.167) between NT embryos (fetus: 2.76±0.40, n =12; membranes: 59.0±10.0, n =11) or in vivo-derived embryos (fetus: 2.60±0.15, n =6; membranes: 41.8±5.2, n =4). However, on Day 75 the weight of the fetus and several of the major organs were heavier from NT embryos. These data suggest that morphological abnormalities involving the fetus and the placenta of cloned pregnancies are manifested after Day 45. [Copyright &y& Elsevier]

Abstract: Many immunological aspects of pregnancy, such as the role played by gamma interferon (IFN-γ) in abortion, are not well understood. Neospora caninum is an intracellular protozoan considered to be among the main causes of abortion in cattle worldwide. The present study analyzes the interaction between IFN-γ production and N. caninum infection in naturally infected pregnant cows. Data were obtained from 126 pregnant cows: 86 seropositive and 40 seronegative for the parasite. Pregnancy diagnosis and blood sample collection were performed on days 40, 90, 120, 150, 180 and 210 post-insemination or until the time of abortion detection. Plasma was tested for antibodies against N. caninum and IFN-γ. Interferon-gamma was detected at some point along the pregnancy in 16 (19%) of the 86 Neospora-seropositive cows yet was undetectable in the 40 seronegative animals. Of the 126 pregnancies examined, 22 (17.5%) ended in abortion. Abortion occurred in 24.4% of seropositive cows (21/86) and in 2.5% of seronegative animals (1/40). Significant (P <0.0001) interaction was observed between Neospora-seropositivity and IFN-γ production. Based on the odds ratio, the risk of abortion was 15.6 times higher in seropositive cows not producing IFN-γ than in seronegative animals, whereas neosporosis had no effect in seropositive cows with IFN-γ production. A significant (P =0.001) negative effect of IFN-γ production on the Neospora titer was furthermore observed in the 65 non-aborting seropositive animals. These results indicate that IFN-γ production affords protection against abortion in Neospora-infected cows and also point to a reduced humoral immune response to N. caninum during gestation in cows producing IFN-γ. [Copyright &y& Elsevier]

Abstract: This research tested the hypothesis that immunological sensitization of mares by skin allografting, followed by the establishment of pregnancy using semen from the skin-graft donor, would give rise to secondary immune responses to the developing horse conceptus, resulting in an earlier demise of the fetally derived endometrial cups. Maiden mares received skin allografts from a stallion homozygous for Major Histocompatibility Complex (MHC) antigens and/or equivalent autografts and were subsequently mated to the skin-graft donor stallion during the next two breeding seasons. Mares that had been immunologically primed to the foreign MHC class I antigens of the skin-graft donor stallion developed strong secondary antibody responses early in their first pregnancies, whereas autografted mares made weak primary antibody responses in their first pregnancies and strong secondary responses in their second pregnancies. In contrast, histological examination of the endometrial cups after surgical pregnancy termination at Day 60 of gestation revealed no discernible differences between allografted and autografted mares, and there were no significant differences in the concentrations and/or duration of secretion of the endometrial cup-specific hormone, equine chorionic gonadotrophin (eCG), between allografted and autografted mares, nor in either group between first and second pregnancies. The vigorous antibody response observed in the pregnant allografted mares supported the first part of our hypothesis, providing evidence of systemic immunological priming. However, there was a lack of an equivalent heightened cellular response to the endometrial cups. These findings provided strong evidence for an asymmetric immune response to the conceptus, characterized by strong humoral immunity and a dampened cellular response. [Copyright &y& Elsevier]

Abstract: Summer heat stress is a main factor related to low conception rate in high producing dairy herds in warm areas worldwide. We assessed the impact of several climate variables on conception rate in high producing dairy cows in northeastern Spain by examining 10,964 inseminations. The temperature–humidity index (THI) was compared with maximum temperature in terms of its efficiency at predicting conception rate. The following data were recorded for each animal: herd, lactation number, insemination number, insemination date, inseminating bull, and AI technician along with climate variables such as mean and maximum temperatures, rainfall, mean and maximum THI for individual time points Days 7 to 1 before insemination, the day of insemination and 1, 2 and 3 days after insemination. Averages were also established for the following periods: from 7 days before insemination to the insemination day, from 3 days before insemination to the insemination day and from the insemination day to 3 days postinsemination. Based on the odds ratios, the likelihood of conception rate increased significantly by factors of 1.48, 1.47, 1.5, and 1.1 for the respective maximum THI classes <70, 71–75, 76–80, and 81–85 only on Day 3 before AI, while on the insemination day, it increased by factors of 1.73, 1.53, 1.11, and 1.3 for the respective maximum THI classes <70, 71–75, 76–80, and 81–85. In a subsequent logistic regression excluding mean and maximum THI, the effectiveness of temperature at predicting conception rate was evaluated. Although high, the fit of the second logistic regression model was slightly lower than that of the full model (P =0.88 versus P =0.98, respectively) and the information provided by the THI model. The likelihood of conception rate increased significantly by factors of 1.5, 1.2, 1.0, 1.0 for the respective maximum temperature classes <20, 21–25, 26–30, 31–35°C on Day 1 after AI. The choice of the THI or temperature to monitor the farm environment would have to depend on the particular farm and situation. In our study conditions, the use of maximum temperature alone gives a new point of view regarding the information provided by the THI variables. [Copyright &y& Elsevier]