Pregnancy Establishment and Diagnosis in Livestock.

This comprehensive review explores the complex processes of reproduction, pregnancy establishment, and pregnancy diagnostic methods in cattle, sheep, goats, swine, horses, and camelids. It provides an overview of the history of pregnancy detection and an in-depth exploration of the physiology of pregnancy in livestock. The detection of conceptus tissue and fluids, conceptus-produced hormones, and maternal responses to conceptus signals, crucial for pregnancy diagnosis, are also discussed in detail, as are emerging methods for pregnancy diagnosis in livestock species. Overall, this review emphasizes the direct impact of pregnancy diagnosis and efficient pregnancy management for profitability of livestock enterprises.

Unsupervised Bayesian classification for models with scalar and functional covariates.

We consider unsupervised classification by means of a latent multinomial variable which categorizes a scalar response into one of the L components of a mixture model which incorporates scalar and functional covariates. This process can be thought as a hierarchical model with the first level modelling a scalar response according to a mixture of parametric distributions and the second level modelling the mixture probabilities by means of a generalized linear model with functional and scalar covariates. The traditional approach of treating functional covariates as vectors not only suffers from the curse of dimensionality, since functional covariates can be measured at very small intervals leading to a highly parametrized model, but also does not take into account the nature of the data. We use basis expansions to reduce the dimensionality and a Bayesian approach for estimating the parameters while providing predictions of the latent classification vector. The method is motivated by two data examples that are not easily handled by existing methods. The first example concerns identifying placebo responders on a clinical trial (normal mixture model) and the other predicting illness for milking cows (zero-inflated mixture of the Poisson model).

Design of a handheld and portable fluorescence imaging system for quantitative detection of pregnancy-specific biomarkers in cattle.

Reproductive management significantly impacts dairy farm productivity, necessitating accurate timely pregnancy detection in cattle. This paper presents a novel handheld and portable fluorescence imaging system designed for quantitative assessment of pregnancy-specific biomarkers, addressing the limitations of current detection methods. The objective was to develop a cost-effective, at-farm solution for detecting pregnancy-specific protein B (PSPB) in bovine plasma samples. The system integrates an imaging module and a custom software application, enabling image capture, data processing, and PSPB concentration determination. Calibration utilizing known PSPB concentrations achieved a 0.6 ng/mL limit of detection. Validation encompassed a comparison with a standard ELISA method using 100 bovine plasma samples; minimal bias and good agreement were observed within the linear range of the calibration curve for both methods. The system offers portability, user-friendliness, and potential for multiplex detection, promising real-time, at-farm reproductive management. This study demonstrates the successful development and validation of a portable fluorescence imaging system, offering an efficient and accurate approach to detecting pregnancy-specific biomarkers in cattle. Its implications extend to improving dairy farm productivity by enabling timely and reliable reproductive management practices.

Effect of a targeted reproductive management program based on automated detection of estrus during the voluntary waiting period on reproductive performance of lactating dairy cows.

The objective of this experiment was to evaluate the effect on reproductive performance of a targeted reproductive management (TRM) program for first postpartum insemination (AI) that prioritized AI at detected estrus (AIE) by providing different intervals for estrus detection based on records of automated estrus alerts (AEA) during the voluntary waiting period (VWP). A secondary objective was to evaluate the association between occurrence of AEA during the VWP and reproductive performance. Lactating Holstein cows (n = 1,260) fitted with neck behavior monitoring sensors for detection of estrus were randomly assigned to a program that used all-timed AI (TAI) for first service (ALL-TAI; n = 632) or a TRM program that prioritized AIE and used TAI only for cows not detected in estrus (TP-AIE; n = 628). Cows in the ALL-TAI treatment received TAI at 76 ± 3 days in milk (DIM) after a Double-Ovsynch protocol. Cows in the TP-AIE treatment were eligible for AIE for 30 ± 3 or 16 ± 3 d after a 49 d VWP if at least one (n = 346) or no (n = 233) AEA were recorded from 15 to 49 DIM. Cows not AIE received TAI after an Ovsynch protocol with progesterone supplementation at 90 ± 3 or 76 ± 3 DIM if the cow had or did not have AEA during the VWP, respectively. Data were analyzed by logistic and Cox's proportional hazard regression. In the TP-AIE treatment, 69.3 % of cows received AIE and more cows with (83.3 %) than without (45.0 %) AEA during the VWP received AIE. Cows in the TP-AIE (69.0 ± 0.7 d) treatment had fewer days from calving to first AI than cows in the ALL-TAI (75.7 ± 0.8 d) treatment. The proportion of cows pregnant by 150 DIM (ALL-TAI = 59.1 % and TP-AIE = 56.0 %) and the hazard ratio (HR) for time to pregnancy (1.0 [95 % confidence interval: 0.9, 1.2]) did not differ between treatments and median days to pregnancy were 102 and 107 for the ALL-TAI and TP-AIE treatments, respectively. Overall, the ALL-TAI (42.3 %) treatment had more first service pregnancies per AI (P/AI) than the TP-AIE (29.0 %) treatment. Cows with AEA during the VWP had greater P/AI (42.5 % vs. 28.9 %), proportion of cows pregnant by 150 DIM (67.4 % vs. 47.0 %), and HR for time to pregnancy (1.6 [1.4, 1.9]) than cows without AEA during the VWP. We conclude that a TRM program that prioritized AIE based on AEA during the VWP led to a similar pregnancy rate and proportion of cows pregnant by mid-lactation than a program that used all-TAI with extended VWP despite fewer P/AI to first service. Also, expression of estrus during the VWP was associated with improved reproductive performance. Thus, AEA during the VWP could be used as a predictor of reproductive potential for TRM of lactating dairy cows.

Reproduction in the era of genomics and automation.

Much progress has been made in the reproductive efficiency of lactating dairy cows across the USA in the past 20years. The standardisation of evaluation of reproductive efficiency, particularly with greater focus on metrics with lesser momentum and less lag-time such as 21-day pregnancy rates (21-day PR), and the recognition that subpar reproductive efficiency negatively impacted profitability were major drivers for the changes that resulted in such progress. Once it became evident that the genetic selection of cattle for milk yield regardless of fertility traits was associated with reduced fertility, geneticists raced to identify fertility traits that could be incorporated in genetic selection programs with the hopes of improving fertility of lactating cows. Concurrently, reproductive physiologists developed ovulation synchronisation protocols such that after sequential treatment with exogenous hormones, cows could be inseminated at fixed time and without detection of oestrus and still achieve acceptable pregnancy per service. These genetic and reproductive management innovations, concurrently with improved husbandry and nutrition of periparturient cows, quickly started to move reproductive efficiency of lactating dairy cows to an upward trend that continues today. Automation has been adopted in Israel and European countries for decades, but only recently have these automated systems been more widely adopted in the USA. The selection of dairy cattle based on genetic indexes that result in positive fertility traits (e.g. daughter pregnancy rate) is positively associated with follicular growth, resumption of ovarian cycles postpartum, body condition score and insulin-like growth factor 1 concentration postpartum, and intensity of oestrus. Collectively, these positive physiological characteristics result in improved reproductive performance. Through the use of automated monitoring devices (AMD), it is possible to identify cows that resume cyclicity sooner after calving and have more intense oestrus postpartum, which are generally cows that have a more successful periparturient period. Recent experiments have demonstrated that it may be possible to adopt targeted reproductive management, utilising ovulation synchronisation protocols for cows that do not have intense oestrus postpartum and relying more heavily on insemination at AMD-detected oestrus for cows that display an intense oestrus postpartum. This strategy is likely to result in tailored hormonal therapy that will be better accepted by the public, will increase the reliance on oestrus for insemination, will improve comfort and reduce labour by reducing the number of injections cows receive in a lactation, and will allow for faster decisions about cows that should not be eligible for insemination.

Effects of feeding rumen-protected methionine pre- and postpartum in multiparous Holstein cows: Health disorders and interactions with production and reproduction.

Study objectives were to evaluate the effects of feeding rumen-protected Met (RPM) in pre- and postpartum total mixed rations (TMR) on health disorders and the interactions of health disorders with lactation and reproductive performance. Multiparous Holstein cows [470; 235 cows at University of Wisconsin (UW) and 235 cows at Cornell University (CU)] were enrolled at approximately 4 wk before parturition and housed in close-up dry cow (n = 6) and replicated lactation pens (n = 16). Pens were randomly assigned to treatment diets (pre- and postpartum, respectively): (1) control (CON): basal diet = 2.30% and 2.09% Met as % of metabolizable protein (MP) (UW) or 2.22% and 2.19% Met as % of MP (CU); (2) RPM: basal diet fed with RPM with 2.83% and 2.58% Met (Smartamine M, Adisseo Inc.; 12 g prepartum and 27 g postpartum), as % of MP (UW) or 2.85% and 2.65% Met (Smartamine M; 13 g prepartum and 28 g postpartum), as % of MP (CU). Total serum Ca was evaluated at the time of parturition and on d 3 ± 1 postpartum. Daily rumination was monitored from 7 d before parturition until 28 d postpartum. Health disorders were recorded during the experimental period until the time of first pregnancy diagnosis (32 d after timed artificial insemination; 112 ± 3 d in milk). Uterine health was evaluated on d 35 ± 3 postpartum. Time to pregnancy and herd exit were evaluated up to 350 d in milk. Treatment had no effect on the incidence of most health disorders and did not alter daily rumination. Cows fed RPM had reduced subclinical hypocalcemia (13.6 vs. 22%; UW only) on day of parturition relative to CON. Percentage of cows culled (13.1 vs. 19.3%) and hazard of herd exit due to culling [hazard ratio = 0.65, 95% confidence interval (CI): 0.42-1.02] tended to be reduced for cows fed RPM compared with CON. Moreover, cows fed RPM had greater milk protein concentration and protein yield overall, although retrospective analysis indicated that RPM only significantly increased protein yield in the group of cows with one or more health disorders (1.47 vs. 1.40 kg/d), not in cows without health disorders (1.49 vs. 1.46 kg/d) compared with CON. Overall, treatment had no effect on pregnancy per timed artificial insemination; however, among cows with health disorders, those fed RPM had reduced time to pregnancy compared with CON (hazard ratio = 0.71, 95% CI: 0.53-0.96). Thus, except for subclinical hypocalcemia on the day of parturition, feeding RPM in pre- and postpartum TMR did not reduce the incidence of health disorders, but our retrospective analysis indicated that it lessened the negative effects of health disorders on milk protein production and time to pregnancy.

Effects of feeding rumen-protected methionine pre- and postpartum on reproductive outcomes of multiparous Holstein cows.

Our primary objective was to evaluate the effect of feeding rumen-protected Met (RPM) in the pre- and postpartum total mixed ration (TMR) on pregnancy per artificial insemination (AI) and pregnancy loss in multiparous Holstein cows. We also evaluated multiple secondary reproductive physiological outcomes before and after AI, including uterine health, ovarian cyclicity, response to synchronization of ovulation, and markers of embryo development and size. A total of 470 multiparous Holstein cows [235 at the University of Wisconsin (UW) and 235 at Cornell University (CU)] were used for this experiment. Experimental treatment diets were applied at the pen level (2 and 4 close-up pens at CU and UW, respectively, and 12 and 6 postfresh pens at CU and UW, respectively); thus, pen was the experimental unit, and cow was the observational unit. Cows were enrolled and randomly assigned to be fed the experimental treatment diets at approximately 4 wk before parturition until 67 d of gestation [147 d in milk (DIM)] after their first service. Close-up dry cow and replicated lactation pens were randomly assigned to treatment diets: RPM, prepartum = 2.83% (UW) and 2.85% (CU), postpartum = 2.58% (UW) and 2.65% (CU); and control (CON), prepartum = 2.30% (UW) and 2.22% (CU), postpartum = 2.09% (UW) and 2.19% (CU; Met as percentage of metabolizable protein). Vaginal discharge and uterine cytology (percentage of polymorphonuclear leucocytes) were evaluated at 35 ± 3 DIM. Cows received timed AI (TAI) at 80 ± 3 DIM after synchronization of ovulation with the Double-Ovsynch protocol. Ovarian cyclicity status, response to synchronization of ovulation, and luteal function were determined by measuring circulating concentrations of progesterone at 35 and 49 ± 3 DIM, 48 and 24 h before TAI, and 8, 18, 22, 25, and 29 d after TAI. Interferon-stimulated gene expression in white blood cells were compared on 18 d after TAI (CU only) and pregnancy-specific protein B concentrations at 22, 25, 29, 32, and 67 d after TAI. Pregnancy status was determined using pregnancy-specific protein B at 25 and 29 d after TAI, and by transrectal ultrasonography at 32, 39, and 67 d after TAI. Embryo and amniotic vesicle size were determined at 32 and 39 d after TAI. Pregnancy per AI (25 d: 64.7 vs. 64.0%, 32 d: 54.3 vs. 55.1% for CON and RPM, respectively) and pregnancy loss (25 to 67 d: 22.6 vs. 19.2% for CON and RPM, respectively) for synchronized cows did not differ. The proportion of cows with purulent vaginal discharge (CON = 7.7 vs. RPM = 4.6%) and cytological endometritis (CON = 20.8 vs. RPM = 23.6%) did not differ. Cyclicity status, ovarian responses to the synchronization protocol, and synchronization rate also did not differ. In addition, fold change for interferon-stimulated genes, concentrations of pregnancy-specific protein B, and embryo size were not affected by treatments. In conclusion, feeding RPM in the pre- and postpartum TMR at the amounts used in this experiment did not affect uterine health, cyclicity, embryo development, or reproductive efficiency in dairy cows.

Effects of feeding rumen-protected methionine pre- and postpartum in multiparous Holstein cows: Lactation performance and plasma amino acid concentrations.

Objectives were to evaluate the effect of feeding rumen-protected methionine (RPM) in pre- and postpartum total mix ration (TMR) on lactation performance and plasma AA concentrations in dairy cows. A total of 470 multiparous Holstein cows [235 cows at University of Wisconsin (UW) and 235 cows at Cornell University (CU)] were enrolled approximately 4 wk before parturition, housed in close-up dry cow and replicated lactation pens. Pens were randomly assigned to treatment diets (pre- and postpartum, respectively): UW control (CON) diet = 2.30 and 2.09% of Met as percentage of metabolizable protein (MP) and RPM diet = 2.83 and 2.58% of Met as MP; CU CON = 2.22 and 2.19% of Met as percentage of MP, and CU RPM = 2.85 and 2.65% of Met as percentage of MP. Treatments were evaluated until 112 ± 3 d in milk (DIM). Milk yield was recorded daily. Milk samples were collected at wk 1 and 2 of lactation, and then every other week, and analyzed for milk composition. For lactation pens, dry matter intake (DMI) was recorded daily. Body weight and body condition score were determined from 4 ± 3 DIM and parturition until 39 ± 3 and 49 DIM, respectively. Plasma AA concentrations were evaluated within 3 h after feeding during the periparturient period [d -7 (±4), 0, 7 (±1), 14 (±1), and 21 (±1); n = 225]. In addition, plasma AA concentrations were evaluated (every 3 h for 24 h) after feeding in cows at 76 ± 8 DIM (n = 16) and within 3 h after feeding in cows at 80 ± 3 DIM (n = 72). The RPM treatment had no effect on DMI (27.9 vs. 28.0 kg/d) or milk yield (48.7 vs. 49.2 kg/d) for RPM and CON, respectively. Cows fed the RPM treatment had increased milk protein concentration (3.07 vs. 2.95%) and yield (1.48 vs. 1.43 kg/d), and milk fat concentration (3.87 vs. 3.77%), although milk fat yield did not differ. Plasma Met concentrations tended to be greater for cows fed RPM at 7 d before parturition (25.9 vs. 22.9 µM), did not differ at parturition (22.0 vs. 20.4 µM), and were increased on d 7 (31.0 vs. 21.2 µM) and remained greater with consistent concentrations until d 21 postpartum (d 14: 30.5 vs. 19.0 µM; d 21: 31.0 vs. 17.8 µM). However, feeding RPM decreased Leu, Val, Asn, and Ser (d 7, 14, and 21) and Tyr (d 14). At a later stage in lactation, plasma Met was increased for RPM cows (34.4 vs. 16.7 µM) consistently throughout the day, with no changes in other AA. Substantial variation was detected for plasma Met concentration (range: RPM = 8.9-63.3 µM; CON = 7.8-28.8 µM) among cows [coefficient of variation (CV) > 28%] and within cow during the day (CV: 10.5-27.1%). In conclusion, feeding RPM increased plasma Met concentration and improved lactation performance via increased milk protein production.

Embryo Mortality Around the Period of Maintenance of the Corpus Luteum Causes Alterations to the Ovarian Function of Lactating Dairy Cows.

Objectives were to identify cows with embryo mortality (EM) around the period of corpus luteum maintenance by interferon tau (IFNT) and to characterize ovarian function in cows that underwent EM. Lactating Holstein cows received artificial insemination (AI) (Day = 0) with semen or extender only. From Day 14 to 42 transrectal ultrasonography was performed daily to monitor ovarian dynamics and uterine contents whereas blood was collected every 48 h to determine ISG15 and MX2 mRNA abundance in blood mononuclear cells (Day 14 to 22 only) and determination of hormone concentrations. Cows were classified in the following reproductive status groups: cyclic (inseminated with extender; n = 15), pregnant (embryo present on Day 42; n = 23), no embryo (n = 23), and EM (n = 14). EM was defined as the presence of an embryo based on interferon-stimulated genes (ISG) mRNA abundance and concentrations of pregnancy-specific protein B (PSPB) above specific cutoff points but no embryo visualized by ultrasonography. Within the EM group, early EM (up to Day 22) was when ISG fold changes were above specific cutoff points from Day 18 to 22 and PSPB below 0.7 ng/ml on and after Day 24, whereas late EM (after Day 22) was when PSPB was above 0.7 ng/ml on or after Day 24 regardless of ISG expression. This experiment provided evidence that the combination of ISG expression patterns and PSPB concentrations is a reasonable method to determine EM around the period of corpus luteum maintenance by IFNT because cows with evidence of EM had patterns of ISG expression more similar to pregnant than cyclic cows or cows with no embryo. Within the EM group, only cows with late EM had delayed luteal regression and longer interovulatory intervals. No major alterations in follicular function were observed after the onset of luteolysis. Our results suggest that embryo development needs to continue beyond 22 days after AI to effectively prevent luteolysis and extend the luteal phase.

Methods for and Implementation of Pregnancy Diagnosis in Dairy Cows.

The first part of this article defines the attributes of the ideal pregnancy test and describes the direct and indirect methods for pregnancy diagnosis in dairy cows that are currently available that have the potential to replace transrectal palpation. Second, this new technology must be practically integrated into a systematic on-farm reproductive management strategy and empirically demonstrated to exceed the status quo of the industry in reproductive performance. Finally a future direction for research and technology in the area of early pregnancy diagnosis in dairy cows is presented, and the overall conclusions of the ideas presented herein are drawn.

Effect of increasing GnRH and PGF2α dose during Double-Ovsynch on ovulatory response, luteal regression, and fertility of lactating dairy cows.

Ovsynch-type synchronization of ovulation protocols have suboptimal synchronization rates due to reduced ovulation to the first GnRH treatment and inadequate luteolysis to the prostaglandin F2α (PGF2α) treatment before timed artificial insemination (TAI). Our objective was to determine whether increasing the dose of the first GnRH or the PGF2α treatment during the Breeding-Ovsynch portion of Double-Ovsynch could improve the rates of ovulation and luteolysis and therefore increase pregnancies per artificial insemination (P/AI). In experiment 1, cows were randomly assigned to a two-by-two factorial design to receive either a low (L) or high (H) doses of GnRH (Gonadorelin; 100 vs. 200 µg) and a PGF2α analogue (cloprostenol; 500 vs. 750 µg) resulting in the following treatments: LL (n = 263), HL (n = 277), LH (n = 270), and HH (n = 274). Transrectal ultrasonography and serum progesterone (P4) were used to assess ovulation to GnRH1, GnRH2, and luteal regression after PGF2α during Breeding-Ovsynch in a subgroup of cows (n = 651 at each evaluation). Pregnancy status was assessed 29, 39, and 74 days after TAI. In experiment 2, cows were randomly assigned to LL (n = 220) or HH (n = 226) treatment as described for experiment 1. For experiment 1, ovulation to GnRH1 was greater (P = 0.01) for cows receiving H versus L GnRH (66.6% [217/326] vs. 57.5% [187/325]) treatment, but only for cows with elevated P4 at GnRH1. Cows that ovulated to GnRH1 had increased (P < 0.001) fertility compared with cows that did not ovulate (52.2% vs. 38.5%); however, no effect of higher dose of GnRH on fertility was detected. The greater PGF2α dose increased luteal regression primarily in multiparous cows (P = 0.03) and tended to increase fertility (P = 0.05) only at the pregnancy diagnosis 39 days after TAI. Overall, P/AI was 47.0% at 29 days and 39.7% at 74 days after TAI; P/AI did not differ (P = 0.10) among treatments at 74 days (LL, 34.6%; HL, 40.8%; LH, 42.2%; HH, 40.9%) and was greater (P < 0.001) for primiparous cows than for multiparous cows (46.1% vs. 33.8%). For experiment 2, P/AI did not differ (P = 0.21) between H versus L treatments (44.2% [100/226] vs. 40.5% [89/220]). Thus, despite an increase in ovulatory response to GnRH1 and luteal regression to PGF2α, there were only marginal effects of increasing dose of GnRH or PGF2α on fertility to TAI after Double-Ovsynch.