Resynchronization with the G6G protocol: A retrospective, observational study of second and later timed artificial inseminations on commercial dairy farms.

We recorded conception rates and estimated pregnancy rates following second and later timed artificial inseminations (TAIs) after hormonal resynchronization on commercial dairy farms, using the so-called G6G protocol (PGF day-0; GnRH 2, 8 days; PGF 15, 16 days, GnRH 17 days; TAI 18 days), and the 5-day Ovsynch protocol or 5DO (GnRH day 0; PGF 5, 6 days; GnRH 7 days; TAI 8 days). In four farms, both protocols were implemented in parallel, and these 1,368 s and later TAIs were used for the protocols' comparison based on logistic regression (544 TAIs in primiparous; 824 in multiparous cows; 1,024 TAIs after G6G [600 TAIs in multiparous and 424 in primiparous]; 344 TAIs after 5DO [224 TAIs in multiparous and 120 in primiparous]; 280 TAIs during the hot season; 1,088 during the cool season). Conception rate (CR) was 31.7% ± 12.0% among all cows, 35.1% ± 10.7% among cows resynchronized with the G6G protocol and 21.8% ± 9.7% among cows resynchronized with the 5DO protocol (p < 0.0001). CR among all cows was lower during the hot season (19.3% ± 8.4%) than during the cool season (34.9% ± 10.6%; p < 0.0001), and similar seasonal results were observed with G6G protocols. Logistic regression showed significant effects on CR in second and later TAIs by protocol (OR = 0.514; 95% CI 0.385-0.686; p < 0.0001) and season (OR = 0.486; 95% CI 0.350-0.676; p < 0.0001). Parity did not influence CR after second and later TAIs (p > 0.1), and no interaction with season or resynchronization protocol was found. Estimated pregnancy rates based on these CR data from both hormonal protocols suggest that G6G can be effectively used for second and later TAIs and highlight the importance of considering protocol and season when designing strategies for second and later timed AIs on dairy farms.

Bull Semen Obtained on Beef Farms by Electroejaculation: Sperm Quality in the First Two Hours of Storing with Different Extenders and Holding Temperatures.

Sperm quality decreases over time, so bull semen may need to be preserved after field collection. However, the effect of handling such semen samples from commercial farms and placing them in very short-term storage has not been elucidated. Therefore, ejaculate from 25 bulls from 1 dairy and 14 beef cattle farms were collected under farm conditions and evaluated for semen quality during the first two hours after collection. Two commercial extenders (AndroMed® and BIOXcell®) and two different storage temperatures (5 °C and room temperature) were used to evaluate the influence on semen quality and sperm kinetics in ejaculates grouped into three evaluation times, based on time since collection (Time 1: <75 min, n = 7; Time 2: 75-105 min, n = 11; and Time 3: 105-120 min, n = 7). Classical semen parameters, sperm motion kinetics by CASA and colony-forming units were assessed. The differences between both extenders in curvilinear and straight-line velocities (VCL and VSL) for the different time groups (Time 2 and Time 3) were statistically significant for p < 0.05. AndroMed® showed lower VSL, straightness and linearity in sperm compared to BIOXcell® (p < 0.05). In conclusion, AndroMed® induced more curvilinear movement, while BIOXcell® stimulated straighter motility.

MOET Efficiency in a Spanish Herd of Japanese Black Heifers and Analysis of Environmental and Metabolic Determinants.

Multiple ovulation and embryo transfer (MOET) systems have been intensively implemented in Japanese Black cattle in Japan and to create Japanese Black herds out of these areas. Environmental conditions influence MOET efficiency. Thus, we describe results of 137 in vivo, non-surgical embryo flushings performed between 2016-2020, in a full-blood Japanese Black herd kept in Spain and the possible effects of heat, year, bull, donor genetic value, and metabolic condition. Additionally, 687 embryo transfers were studied for conception rate (CR) and recipient related factors. A total of 71.3% of viable embryos (724/1015) were obtained (5.3 ± 4.34/flushing). Donor metabolites did not affect embryo production (p > 0.1), although metabolite differences were observed over the years, and by flushing order, probably related to the donor age. CR was not affected by embryo type (fresh vs. frozen), recipient breed, and whether suckling or not suckling (p > 0.1). CR decreased significantly with heat (44.3 vs. 49.2%; (p = 0.042)) and numerically increased with recipient parity and ET-number. Pregnant recipients showed significantly higher levels of cholesterol-related metabolites, glucose, and urea (p < 0.05). Therefore, adequate MOET efficiency can be achieved under these conditions, and heat stress should be strongly avoided during Japanese Black embryo transfers. Moreover, recipients' metabolites are important to achieve pregnancy, being probably related to better nutrient availability during pregnancy.

Effects on Synchronization and Reproductive Efficiency of Delaying the Removal of the Intravaginal Progesterone Device by 24 h in the 5d Co-Synch Protocol in Heifers.

In beef herds, increasing animal welfare, improving reproductive performance and easing animal management are key goals in farm economics. We explored whether delaying the removal of the intravaginal progesterone device by 24 h in heifers synchronized with a 5d Co-synch 72-h protocol could improve reproductive efficiency of fixed-time artificial insemination (FTAI). In experiment 1, we examined the total synchronization rate (TSR) in cycling Holstein heifers. Heifers (13.4 ± 0.69 mo.) were randomly assigned to the standard 5d Co-synch 56-h protocol (5dCo56; n = 10), 5d Co-synch 72-h (5dCo72; n = 17), or the modified 5d Co-synch 72-h protocol, in which removal of the progesterone device was delayed by 24 h (6dCo48; n = 19). In experiment 2, 309 cycling beef heifers on 18 commercial farms were subjected to the 5d Co-synch 72-h or 6-d Co-synch 48-h protocol and conception rate (CR) studied. In experiment 1, the three protocols led no differences on TSRs of 80.0% (5dCo56), 88.2% (5dCo72), and 89.5% (6dCo48). In experiment 2, the CR from the beef heifers, observed during two consecutive reproductive seasons did not differ: 59.7% for 5dCo72 and 62.0% for 6dCo48 (p = 0.907). Therefore, delaying removal by 24 h provides satisfactory results without reducing reproductive efficiency of heifers.

Effects of Extender Type, Storage Time, and Temperature on Bull Semen Parameters.

Seminal parameters can be evaluated in situ, or samples can be delivered to a diagnostic centre. How storage conditions affect ejaculates up to evaluation is unclear. We assessed, in 25 commercial bulls electroejaculated in the field, the impact of time until evaluation (0-2 h, 4-6 h, and 24 h post-ejaculation), holding temperature (5 °C vs. room temperature), and extender (AndroMed®, BIOXcell® or INRA96®) on semen quality. Acrosome integrity, sperm viability and morphology, CASA-total and progressive motility, pH, and colony-forming units were assessed. Semen quality was preserved for up to 4-6 h post-ejaculation, except for INRA96® at 5 °C. Regardless of extender or temperature, motility decreased from 4 to 6 h up to 24 h, with the best values obtained with BIOXcell® at 5 °C. pH differed from 4 to 6 h up to 24 h, acidifying when stored at room temperature. Microbiological load was stable over time with AndroMed® and BIOXcell®, and increased at room temperature with INRA96®. Our results suggest that AndroMed® and BIOXcell® can preserve semen quality for up to 6 h, either at 5 °C or room temperature, while INRA96® only at room temperature. These results help to fix adequate protocols for short-term storage and shipment of bovine semen collected under field conditions.

Effect of Extender, Storage Time and Temperature on Kinetic Parameters (CASA) on Bull Semen Samples.

CASA kinetic parameters are often evaluated in a diagnostic centre. How storage conditions affect ejaculates up to evaluation is unclear. We assessed, in 25 commercial bulls electroejaculated in the field, the impact of time until evaluation (0-2 h, 4-6 h, and 24 h post-ejaculation), holding temperature (5 °C vs. room temperature), and extender (AndroMed®, BIOXcell® or INRA96®) on CASA kinetic parameters. Total and progressive motility, VCL, VAP, VCL, ALH, BCF, STR, LIN, and WOB were assessed. CASA kinetic parameters were preserved for up to 4-6 h post-ejaculation, except for AndroMed®. Regardless of extender or temperature, motility decreased from 4-6 h up to 24 h, with the best values obtained with BIOXcell® at 5 °C. Our results suggest that BIOXcell® can preserve sperm motility for up to 6 h, either at 5 °C or room temperature, and also INRA96® at room temperature, with motility assessments and the percentage of the most rapid sperms being the lowest with INRA96® at 5 °C. The kinetic parameters decreased when analyses were performed at 24 h. Therefore, we suggest evaluating seminal quality as soon as possible, before 6 h after collection. These results help to fix adequate protocols for the short-term storage and shipment of bovine semen collected under field conditions.

The Effect of Stress on Reproduction and Reproductive Technologies in Beef Cattle-A Review.

Researchers have contributed by increasing our understanding of the factors affecting reproduction in beef, mainly physical health and nutrition aspects, which have been main concerns during decades. Animal welfare is of outmost relevance in all animal production systems and it is strongly associated to stress. Stress responses involve endocrine, paracrine and neural systems and the consequences of this stress on the reproductive efficiency of specifically, beef cattle and bulls, need to be highlighted. We, therefore, describe the fundamentals of stress and its quantification, focusing in beef herds, reviewing the highly valuable pieces of research, already implemented in this field. We examine major factors (stressors) contributing to stress in beef cattle and their effects on the animals, their reproductive performance and the success of reproductive biotechnologies. We include terms such as acclimatization, acclimation or temperament, very relevant in beef systems. We examine specifically the management stress due to handling, social environment and hierarchy or weaning effects; nutritional stress; and thermal stress (not only heat stress) and also review the influence of these stressors on reproductive performance and effectiveness of reproductive biotechnologies in beef herds. A final message on the attention that should be devoted to these factors is highlighted.

Pregnancy Loss (28-110 Days of Pregnancy) in Holstein Cows: A Retrospective Study.

The objective of this retrospective study was to investigate the prevalence of pregnancy loss (PL; between 28-110 pregnancy days) and its relationship with factors: farm, year (2015-2018), season, artificial insemination (AI)-rank, parity, AI-type (fixed-time vs. oestrus-AI), previous PL, days in milk (DIM), fixed-time-AI protocol, AI-technician, bull, and semen-type (sexed vs. conventional). Data of 19,437 Holstein cattle AIs from eight Spanish farms were studied. Overall conception rate was 34.3% (6696/19,437) and PL 12.3% (822/6696). The PL was more likely to occur in primiparous (10.8%, odds ratio (OR) = 1.35; p = 0.04) and multiparous (15.3%; OR = 2.02, p < 0.01) than in heifers (PL = 6.9%, reference). Pregnancies achieved with AI after observed oestrus and natural breedings were associated with less PL than pregnancies after fixed-time-AI (12.7 vs. 11.9%; OR = 0.12, p = 0.01). First AIs related to higher PL than ≥2nd AIs (PL = 13.8% vs. 11.2; OR = 0.73, p < 0.01). The factors season, fixed-time-AI protocol, DIM, bull, AI-technician, or type of semen were not significantly associated with PL. Therefore, farmers and consultants should adapt their preventive strategies relating to PL, particularly, to the parity of the cattle.

Thoracoscopy as a safe and effective technique for exploring calves affected with bovine respiratory disease.

BACKGROUND: Bovine respiratory disease (BRD) is one of the leading causes of economic losses in the beef and dairy industry. Reliable antemortem tools for diagnosing BRD would improve the efficacy of treatment and reduce costs. Here we examined whether the relatively simple technique of thoracoscopy can support BRD diagnosis under field conditions. We also compared various equipment set-ups in order to optimize the safety and efficacy of the procedure. A total of 24 thoracoscopic procedures were performed in 17 calves diagnosed with BRD and in 2 healthy control calves. Rigid and flexible endoscopes and industrial videoscopes were tested using various insertion approaches. The suitability of the technique was assessed in terms of duration, volume of air extracted, visualization score, and image quality. Safety was assessed in terms of rectal temperature, body weight, breaths/min, presence of fibrinogen, pain score, recovery time, intraoperative complications and risk of laceration or threatening collapse. RESULTS: Insertion of a flexible endoscope via a right, dorso-caudal approach at the 5th intercostal space allowed complete examination of the right lung in 15 min, as well as identification of main lung lesions and adherences in calves with BRD, without compromising calf welfare. While the dorso-caudal approach was optimal, it was associated with substantial discomfort when rigid endoscopes were used, minimal complications or mortality due to thoracoscopy were observed up to 28 days after the procedure. Videoscopes were as safe and easy to use as endoscopes, but endoscopes provided better image quality. CONCLUSION: This study provides the first field evidence that thoracoscopy can be safe to explore BRD-diseased calves. These results justify a larger study to rigorously assess the diagnostic performance of the technique.