Theriogenology 

Abstract: This paper reports a new method for synchronizing the time of ovulation in cattle using GnRH and PGF(2alpha). In Experiments 1 and 2, lactating dairy cows (n=20) ranging from 36 to 280 d postpartum and dairy heifers (n=24) 14 to 16 mo old were treated with an intramuscular injection of 100 mug GnRH at a random stage of the estrous cycle. Seven d later the cattle received PGF(2alpha) to regress corpora lutea (CL). Lactating cows and heifers received a second injection of 100 mug GnRH 48 and 24 h later, respectively. Lactating cows were artificially inseminated 24 h after the second GnRH injection. Ovarian morphology was monitored daily by trans-rectal ultrasonography from 5 d prior to treatment until ovulation. In Experiment 3, the flexibility in the timing of hormonal injections with this synchronization protocol was evaluated by randomly assigning 66 lactating dairy cows to 3 different treatment groups. Lactating cows received the injection of PGF(2alpha) 48 (Group 1), 24 (Group 2), and 0 h (Group 3) prior to the second injection of GnRH, which was administered at the same time in each group to ensure the second injection of GnRH was given when follicles were at a similar stage of growth. In Experiments 1 and 2, the first injection of GnRH caused ovulation and formation of a new or accessory CL in 18 20 cows and 13 24 heifers. In addition, this injection of GnRH initiated or was coincident with initiation of a new follicular wave in 20 20 lactating cows and 18 24 heifers. Corpora lutea regressed after PGF(2alpha) in 20 20 cows and in 18 24 heifers. All cows and 18 24 heifers ovulated a newly formed dominant follicle between 24 and 32 h after the second injection of GnRH. Ten of 20 cows conceived to the timed artificial insemination. In Experiment 3, the conception rate in Groups 1 and 2 were greater than in Group 3, (55 and 46 % vs 11%, respectively). In summary, this protocol could have a major impact on managing reproduction in lactating dairy cows, because it allows for AI to occur at a known time of ovulation and eliminates the need for detection of estrus.

Abstract: A procedure to obtain high and repeatable fertilization frequencies for bovine in vitro fertilization (IVF) with frozen-thawed sperm was developed. IVF frequency of in vitro matured oocytes was increased by a swimup sperm separation procedure (P=0.01) or treatment of sperm with the glycosaminoglycan heparin (P=0.0001), but the two factors did not interact (P=0.23). Heparin was the most important factor in increasing IVF frequencies. The fertilization frequency was not affected by the batch of oocytes used (P=0.38), but bull effects were present (P<0.05). Within a bull, the IVF system was highly repeatable and varied between trials no more than +/- 12% in fertilization frequency with an overall fertilization frequency of 299 379 (79%) on four trials over four bulls. In vivo matured oocytes fertilized in vitro were transferred to ewe or heifer oviducts. Morulae or blastocysts were recovered from ewes after four to five days, while conceptuses were present in the bovine after 25 days (diagnosed by ultrasound). Embryonic development from the IVF system either pre- or postimplantation was normal.

Abstract: Uterine function is often compromised in cattle by bacterial contamination of the uterine lumen after parturition, and pathogenic bacteria often persist, causing uterine disease, a key cause of infertility in cattle. However, the definition or characterization of uterine disease frequently lacks precision or varies among research groups. The aim of the present paper was to provide clear clinical definitions of uterine disease that researchers could adopt. Puerperal metritis should be defined as an animal with an abnormally enlarged uterus and a fetid watery red-brown uterine discharge, associated with signs of systemic illness (decreased milk yield, dullness or other signs of toxemia) and fever > 39.5 degrees C, within 21 days after parturition. Animals that are not systemically ill, but have an abnormally enlarged uterus and a purulent uterine discharge detectable in the vagina, within 21 days post partum, may be classified as having clinical metritis. Clinical endometritis is characterised by the presence of purulent (> 50% pus) uterine discharge detectable in the vagina 21 days or more after parturition, or mucuopurulent (approximately 50% pus, 50% mucus) discharge detectable in the vagina after 26 days post partum. In the absence of clinical endometritis, a cow with subclinical endometritis is defined by > 18% neutrophils in uterine cytology samples collected 21-33 days post partum, or > 10% neutrophils at 34-47 days. Pyometra is defined as the accumulation of purulent material within the uterine lumen in the presence of a persistent corpus luteum and a closed cervix. In conclusion, we have suggested definitions for common postpartum uterine diseases, which can be readily adopted by researchers and veterinarians.

Abstract: Lameness has contributed to reproductive inefficiency and increased the risk of culling in dairy cows. We developed a 5-point lameness scoring system that assessed gait and placed a novel emphasis on back posture. Our objective was to determine if this system predicted future reproductive performance and the risk of culling. The study was conducted at a commercial dairy farm with a history of declining reproductive efficiency and an increasing prevalence of lameness. A total of 66 primipara and pluripara calved, received an initial lameness score, and completed their 60-d voluntary waiting period. The overall prevalence of lameness (mean lameness score ‘2) was 65.2%. Scoring continued at 4-wk intervals and ceased with conception or culling. The percentage of cows confirmed pregnant and culled was 77.3 and 22.7, respectively. For each reproductive endpoint, a 2 x 2 table was constructed with lameness score >2 as the positive risk factor and either performance greater than the endpoint mean or being culled as the positive disease or condition. Positive and negative predictive values, relative risk, Chisquare statistic and regression analysis were used to evaluate the data. The positive predictive values for days to first service, days open, breeding herd days, services per pregnancy and being culled were 58, 68, 65, 39 and 35%, respectively. Similarly, the negative predictive values were 79, 96, 100, 96 and lOO%, respectively. Except for one reproductive endpoint, the total number of services, all linear regressions were significant at P 2 predicted that a cow would have extended intervals from calving to first service and to conception, spend or be assigned to (explained herein) more total days in the breeding herd, require more services per pregnancy and be 8.4 times more likely to be culled. We believe that this lameness scoring system effectively identifies lame cows. Observation of the arched-back posture in a standing cow (>LS 3) should trigger corrective interventions. 0 1997 by Elsevier Science Inc

Abstract: We describe a bovine embryo culture system that supports repeatable high development in the presence of serum or BSA as well as under defined conditions in the absence of those components. In the first experiment, embryo development in SOF with amino acids (SOFaa), sodium citrate (SOFaac) and myo-inositol (SOFaaci) and with BSA or polyvinyl alcohol (PVA) was compared with that in a M199 granulosa cell co-culture (M199 co-culture). Subsequently, development and cell numbers of blastocysts cultured under defined conditions in SOFaaci with PVA (SOFaaci-PVA), or under undefined conditions in SOFaaci with 5% cow serum (SOFaaciCS) or M199 co-culture were compared. The repeatability of culture results in SOFaaci-CS was checked by weekly replicates (n = 30) spread over 11 months. The viability of embryos developed in SOFaaci-PVA was estimated by transfer of morphologically good blastocysts (n = 10) to synchronized recipients. In the second experiment, the effect of omitting CS or BSA from IVM and IVM-IVF on subsequent embryo development in SOFaaci-PVA or in SOFaaci-CS was investigated. Blastocyst development in SOFaa-PVA, SOFaac-PVA, SOFaa-BSA and M199 was 16 ± 3, 23 ± 2ab, 30 ± 8a and 36 ± 7a%, respectively (Pab < 0.05). Additional inclusion of myoinositol resulted in 42 ± 1a% blastocysts in SOFaaci-PVA vs 19 ± 3b% in SOFaac-PVA, 47 ± 7a% in SOFaac-BSA, and 36 ± 7a% in M199 co-culture, respectively (Pab < 0.01). In 30 replicates, the average cleavage and blastocyst rates of oocytes in SOFaaci-CS were 87 ± 4 and 49 ± 5%, respectively. Five normal calves were produced after transfer of 10 blastocysts developed in defined culture medium (i.e., SOFaaci-PVA). Defined IVM or IVM-IVF (i.e., in absence of CS and BSA) reduced cleavage rates (83 ± 3 and 55 ± 3% vs 90 ± 1% in presence of CS; P < 0.01). Subsequent embryo development in SOFaaci-CS was not affected in either of these defined conditions. However, cleavage and blastocyst rates under completely defined IVP conditions were 54 ± 7 and 19 ± 4%, respectively. It was concluded that under defined culture conditions, addition of citrate and myo-inositol improved blastocyst development to rates comparable to those obtained with serum, BSA or co-culture and that the quality of blastocysts was not affected by the absence of serum or BSA. However, serum was essential during IVM/IVF for normal fertilization and subsequent high blastocyst development.