Influence of sub-clinical endometritis on early pregnancy predictors and proinflammatory cytokines in circulating immune cells in dairy cows.

In Experiment 1, PBMC were isolated from cows considered healthy or with SCE (n=6/group) on Days 0 (estrus) and 7 (diestrus) of a synchronized estrous cycle. In Experiment 2, on D21 (D0 was defined as the day of Fixed Timed Artificial Insemination (FTAI), cows were evaluated by ultrasonography to assess luteal blood perfusion and PBMC were isolated. On D32, cows were classified into: healthy pregnant (n=7), pregnant with SCE (n=4), healthy non-pregnant (n=8), and non-pregnant with SCE (n=10). Gene expression of ISGs (ISG15, OAS1, MX1 and IFI6) and proinflammatory cytokines (IL1-ß, TNF-α and IFN-γ) were determined. Expression of ISG15, MX1, IFI6, TNF-α and IFN-γ did not differ between SCE and healthy cows and between Days 0 and 7. Expression of OAS1 and IL1-ß were higher (P=0.02) on Day 7 than Day 0, regardlees of the SCE presence. In Exp.2, ISG15 abundance was 2.5-fold greater (P=0.0008), TNF-α was 2.2-fold greater (P=0.05), and IL1-ß tended (P=0.06) to be 2.4-fold higher in pregnant than non-pregnant cows. Luteal blood perfusion was greater (P=0.01) in pregnant animals. In conclusion, OAS1 and IL1-ß are transcripts upregulated in PBMC at diestrus, regardless of SCE occurrence. Proinflammatory cytokines are not affected by SCE occurrence, but IL1-ß and TNF-α are upregulated in pregnant animals on D21 of pregnancy. ISG15 abundance is a good pregnancy predictor, regardless SCE presence.

Impact of 17ß-estradiol administration at the moment of timed-AI in Nelore cows with small dominant follicle or not showing estrus.

We aimed to evaluate the effects of administering estradiol (E-17ß) at the moment of timed-AI (TAI) on uterine gene expression, estrous expression rate (EER), and pregnancy rate (P/TAI) in Nelore cows with a small dominant follicle (DF) or not showing estrus at TAI. In Experiments 1 and 2 (Exp1, Exp2) cows were submitted to a P4/E-17ß-based protocol (day 0) for synchronization of ovulation. On day 7, devices were removed, cows received 1 mg E-17ß cypionate and 12.5 mg dinoprost. On day 9, cows with DF < 11.5 mm in diameter were split into different groups. In Exp1 (n = 16/group): Control (no treatment), E-2 (2 mg E-17ß) and E-4 (4 mg E-17ß). In Exp2: Control (n = 12); E-2 (n = 14); GnRH (0.1 mg gonadorelin acetate, n = 13); and E-2+GnRH (association of GnRH and E-17ß, n = 13). Between days 9 and 11, endometrial thickness (ET), time of ovulation detection, and EER were recorded. In Exp1, a uterine cytological sample was collected 4 h after treatment to evaluate the transcript expression of receptors for E-17ß (ESR1 and ESR2), oxytocin (OXTR), and P4 (PGR). In Experiment 3 (Exp3), 3829 suckled cows were submitted to a P4/E-17ß-based protocol for TAI. On day 9, devices were removed and cows received 1 mg E-17ß cypionate and 0.4 mg sodium cloprostenol. On day 11, TAI was performed and cows that did not demonstrate estrus received 0.1 mg gonadorelin acetate, and were allocated into two groups: GnRH (n = 368) and E-2+GnRH (2 mg E-17ß; n = 363). In Exp1, plasma E-17ß concentrations increased at 4 h after treatment in a dose-dependent manner but reduced at 12 h. The E-17ß-treated cows had greater transcript abundance for OXTR and lesser for ESR1 and ESR2, and the ET was reduced 12 h after treatment (P < 0.05). No significant difference (P > 0.1) was observed between the E-17ß doses in estrus or ovulation rate. In Exp2, the interval from treatment to ovulation was longer (P < 0.05) in the E-17ß group. GnRH-treated cows showed higher ovulation rates (89 vs. 35 %) compared to cows not treated with GnRH, as E-17ß-treated cows (P < 0.01) had a lower ovulation rate compared to those not receiving E-17ß (44 vs. 78 %). In Exp3, P/TAI was 55 % for cows in estrus. For those not showing estrus, no difference (P > 0.1) in P/TAI was observed between GnRH (34 %) and E-2+GnRH (31 %) groups. Cows with a DF ≥ 11 mm (n = 192) had a greater (P < 0.05) P/TAI (49 %) than those with DF < 11 mm (n = 377; 29 %). In conclusion, E-17ß administration in the moment of TAI modulates the mRNA expression of uterine receptors in cows with a small DF but does not impact the P/TAI compared with GnRH treatment in suckled Nelore not showing estrus previous to TAI.

Accuracy of early pregnancy diagnosis and determining pregnancy loss using different biomarkers and machine learning applications in dairy cattle.

This study aimed to compare the accuracy of IFN-τ stimulated gene abundance (ISGs) in peripheral blood mononuclear cells (PBMCs), CL blood perfusion by Doppler ultrasound (Doppler-US), plasma concentration of P4 on Day 21 and pregnancy-associated glycoproteins (PAGs) test on Day 25 after timed-artificial insemination (TAI) for early pregnancy diagnosis in dairy cows and heifers. Holstein cows (n = 140) and heifers (n = 32) were subjected to a hormonal synchronization protocol and TAI on Day 0. On Day 21 post-TAI, blood samples were collected for PBMC isolation and plasma concentration of P4. The CL blood perfusion was evaluated by Doppler-US. Plasma samples collected on Day 25 were assayed for PAGs. The abundance of ISGs (ISG15 and RSAD2) in PBMCs was determined by RT-qPCR. Pregnancy was confirmed on Days 32 and 60 post-TAI by B-mode ultrasonography. Statistical analyses were performed by ANOVA using the MIXED procedure and GLIMMIX in SAS software. The pregnancy biomarkers were used to categorize the females as having undergone late luteolysis (LL); early embryonic mortality (EEM); late embryonic mortality (LEM); or late pregnancy loss (LPL). The abundance of ISGs, CL blood perfusion by Doppler-US, and concentrations of P4 on Day 21, and PAGs test on Day 25 were significant (P < 0.05) predictors of early pregnancy in dairy cows and heifers. Dairy cows had a greater (P = 0.01) occurrence of LL than heifers, but there was no difference (P > 0.1) for EEM, LEM, and LPL in heifers compared to cows. Cows with postpartum reproductive issues had a greater (P = 0.008) rate of LEM and a lesser (P = 0.01) rate of LPL compared to cows without reproductive issues. In summary, the CL blood perfusion by Doppler-US had the highest accuracy and the least number of false negatives, suggesting it is the best predictor of pregnancy on Day 21 post-TAI. The PAGs test was the most reliable indicator of pregnancy status on Day 25 post-TAI in dairy heifers and cows. The application of machine learning, specifically the MARS algorithm, shows promise in enhancing the accuracy of predicting early pregnancies in cows.

Effects of Maternal Nutrition on Female Offspring Weight Gain and Sexual Development.

Maternal nutrition during pregnancy influences postnatal life of animals; nevertheless, few studies have investigated its effects on the productive performance and reproductive development of heifers. This study evaluated the performance, reproductive development, and correlation between reproduction × fat thickness and performance × ribeye area (REA) traits of heifers. We also performed an exploratory genomic association during the rearing period in heifers submitted to fetal programming. The study comprised 55 Nellore heifers born to dams exposed to one of the following nutritional planes: control, without protein-energy supplementation; PELT, protein-energy last trimester, protein-energy supplementation offered in the final third of pregnancy; and PEWG, protein-energy whole gestation, protein-energy supplementation upon pregnancy confirmation. Protein-energy supplementation occurred at the level of 0.3% live weight. After weaning, heifers were submitted to periodic evaluations of weight and body composition by ultrasonography. From 12 to 18 months, we evaluated the reproductive tract of heifers to monitor its development for sexual precocity and ovarian follicle population. The treatments had no effect (p > 0.05) on average daily gain; however, the weight of the animals showed a significant difference over time (p = 0.017). No differences were found between treatments for REA, backfat, and rump fat thickness, nor for puberty age, antral follicular count, and other traits related to reproductive tract development (p > 0.05). The correlation analysis between performance traits and REA showed high correlations (r > 0.37) between REA at weaning and year versus weight from weaning until yearling; however, no correlation was found for reproductive development traits versus fat thickness (p > 0.05). The exploratory genomic association study showed one single-nucleotide polymorphism (SNP) for each treatment on an intergenic region for control and PEWG, and the one for PELT on an intronic region of RAPGEF1 gene. Maternal nutrition affected only the weight of the animals throughout the rearing period.

A Randomized Clinical Trial Evaluating the Effect of an Oral Calcium Bolus Supplementation Strategy in Postpartum Jersey Cows on Mastitis, Culling, Milk Production, and Reproductive Performance.

The objectives of this study were to evaluate the effects of a postpartum oral calcium supplementation strategy in multiparous Jersey cows on (1) the odds of clinical mastitis in the first 60 days in milk (DIM); (2) the odds of culling up to 60 DIM; (3) the risk of pregnancy in the first 150 DIM; (4) milk production in the first 15 weeks of lactation. A randomized clinical trial was performed in a dairy herd located in west Texas, United States. A total of 809 cows were used in the final analyses. Overall, postpartum oral calcium supplementation did not influence milk production, reproductive performance, or culling. Among second parity cows, oral calcium supplementation tended to decrease the odds of clinical mastitis in the first 60 DIM compared to controls; however, no differences were observed for cows in parities three and greater. To date, data evaluating the effect of postpartum oral calcium supplementation in multiparous Jersey cows are limited. In our study, oral calcium supplementation tended to reduce clinical mastitis in second parity cows. No positive benefits based on the reduction of culling, and improvement of milk production and reproductive performance were evident for the herd included in this study.

Biochemical profile in dairy cows with artificial induction of lactation / Perfil bioquímico de vacas leiteiras submetidas a indução artificial de lactação

The objective of this study was to determine the biochemical profile of dairy cows with induced lactation. For comparison, another group of normally calved cows was used as control. Lactation was induced in multiparous Holstein cows (n=10) with two norgestomet implants (3mg each implant) on day 1. The testing continued with intramuscular norgestomet (3mg/animal) on days 1, 3, 5, 7, 9, 11, 13 and 15. On days 1, 9, 16 to 18 and then every 14 days, bSTr (500mg/animal) was added. On day 16, the intravaginal implant was removed and intramuscular prostaglandin F2α (0.530mg/animal) and intramuscular estradiol benzoate (5mg/animal) were added. On days 16 to 18 dexamethasone (10mg/animal) was added, and from days 18 to 20 intramuscular metoclopramide (100mg/animal) was added. Milking began on day 19 of the induction. Blood was collected for a biochemical profile after 21 days in milk. It was found that urea and triglyceride concentrations were significantly higher in the induced cows (P<0.05). Therefore, it was concluded that the animals that had lactation induced did not present disorders related to the biochemical profile indicating that the hepatic function, renal function and lipidogram of the animals were not affected by the use of the drugs to induce lactation.(AU)

O objetivo deste estudo foi determinar o perfil bioquímico de vacas leiteiras com submetidas a indução de lactação. Para comparação, outro grupo de vacas que apresentaram parto normal foi usado como controle. A lactação foi induzida em vacas Holandesas (n=10) utilizando dois implantes de norgestomet (3mg cada implante) no dia 1. O protocolo continuou com a aplicação de norgestomet intramuscular (3mg / animal) nos dias 1, 3, 5, 7, 9, 11, 13 e 15. Nos dias 1, 9, 16 a 18 e depois a cada 14 dias, foi adicionado bSTr (500mg / animal). No dia 16, o implante intravaginal foi removido e adicionou-se prostaglandina F2a intramuscular (0,530 mg / animal) e benzoato de estradiol intramuscular (5mg / animal). Nos dias 16 a 18 foi adicionada dexametasona (10mg / animal) e dos dias 18 a 20 foi adicionada metoclopramida intramuscular (100 mg / animal). A ordenha começou no dia 19 da indução. O sangue foi coletado para mensuração do perfil bioquímico após 21 em leite. Verificou-se que as concentrações de ureia e triglicérides foram significativamente maiores nas vacas induzidas (P<0,05). Portanto, concluiu-se que os animais que tiveram a lactação induzida não apresentaram distúrbios relacionados ao perfil bioquímico, indicando que a função hepática, a função renal e o lipidograma dos animais não foram afetados pelo uso das drogas para induzir a lactação.(AU)