An inflammatory response is induced in the reproductive tract by deposition of semen during natural mating. This response might facilitate establishment and maintenance of pregnancy and alter the phenotype of the offspring by modifying the microenvironment of the reproductive tract. Here, we hypothesized that intrauterine infusion of 0.5 mL of seminal plasma at the time of artificial insemination (AI) in first-service lactating Holstein cows will improve pregnancy success after insemination. Cows were inseminated (511 primiparous cows inseminated with X-sorted semen, 554 multiparous cows inseminated with X-sorted semen, and 627 multiparous cows inseminated with conventional semen) using the Double-Ovsynch protocol. Cows were randomly assigned to receive intrauterine infusion of either 0.5 mL of seminal plasma or saline immediately after AI. There was no overall effect of seminal plasma infusion on the percentage of inseminated cows diagnosed pregnant at d 32 or 60 after AI, pregnancy loss, or percent of inseminated cows calving. If cows were inseminated with conventional semen, seminal plasma reduced pregnancies at d 32 and tended to reduce calvings. There was no effect of seminal plasma if cows were inseminated with X-sorted semen. Seminal plasma infusion increased the birth weight of heifer calves born using X-sorted semen but not conventional semen. These results do not support a beneficial effect of seminal plasma on pregnancy success after AI, but exposure to seminal plasma may program fetal development to affect phenotype at birth.

Once it enters the uterus at d 4 to 5 after ovulation, the preimplantation bovine embryo is controlled in its development by regulatory signaling molecules from the mother called embryokines. Here, several cell-signaling molecules whose genes are expressed in the endometrium during d 5 to 7 after estrus were tested for the ability to affect the competence of the embryo for further development and the characteristics of the resultant blastocysts. Molecules tested were Cnatriuretic peptide (CNP), IL-8, bovine morphogenetic protein 4 (BMP-4), IL-6, and leukemia inhibitory factor (LIF). None of the cell-signaling molecules tested improved the competence of the embryo to become a blastocyst; in fact, BMP-4 decreased development. All molecules modified attributes of the blastocyst formed in culture. In particular, CNP increased the number of cells in the ICM, whereas IL-8 decreased inner cell mass cell numbers and tended to increase the proportion of blastocysts that were hatching or hatched. In addition, BMP-4 decreased the proportion of blastocysts that were hatching. Interleukin-6 and, to a lesser extent, LIF activated the Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in the inner cell mass, and LIF increased the percent of cells in the blastocyst that were positive for both NANOG and phosphorylated (activated) STAT3. In conclusion, our results indicate that CNP, IL-8, IL-6, LIF, and BMP-4 can modify embryonic development of the cow in a manner that affects characteristics of the resultant blastocyst. Further research is required to understand how these changes in characteristics of the blastocyst would affect competence of the embryo to establish and maintain pregnancy.

No abstract

El objetivo fue evaluar el impacto de factores asociados al manejo de la madre y la calidad del calostro sobre la crianza de reemplazos bovinos durante la lactancia, en el sistema de producción de leche a pequeña escala. En 13 unidades de producción se obtuvo información del crecimiento corporal, la alimentación con calostro e información asociada a la madre de 220 becerras Holstein. Las variables de interés fueron: morbilidad, concentración de inmunoglobulinas en el  calostro  (CIC; <110 mg/ml),  concentración de proteína  sérica (CPS; <6.6 g/dl), ganancia diaria de peso (GDP; <0.650 kg/día) y ganancia diaria de altura (GDA;  <0.222 cm/día).  Los factores  de  estudio  fueron: peso corporal  al  nacimiento (PCN; <42 kg),  altura al nacimiento (AN; <82 cm), calostro consumido  el primer día (CCPD; <4 l), calostro consumido el primer día/kg de PV (CPDPV), condición corporal al parto (CCP; <3), duración del periodo seco (DPS; >68 días), vacas primíparas (VP) y vacas sin dieta de reto (VSDR). Para determinar el impacto de los factores de estudio sobre los eventos de interés, se obtuvo la razón de momios a partir de análisis de regresión logística múltiple. Los factores identificados para morbilidad fueron CCPD y VP (P<0.1). Los factores para CIC fueron la DPS y VSDR (P<0.1), mientras que para la GDP lo fue VSDR (P<0.1). Finalmente, los factores para GDA fueron VSDR, PCN, AN y CPDPV (P<0.1). Estos resultados sugieren que la nutrición de la madre durante la gestación tardía tiene un impacto importante sobre la salud y el desarrollo corporal de las becerras durante la lactancia. Estudios adicionales deberán determinar los efectos a largo plazo.

The use of assisted reproductive technologies (ART) in cattle can result in large/abnormal offspring syndrome (LOS/AOS) which is characterized by macrosomia. LOS can cause dystocia and lead to the death of dam and calf. Currently, no test exists to identify LOS pregnancies. We hypothesized that fetal ultrasonography and/or maternal blood markers are useful to identify LOS. Bovine fetuses were generated by artificial insemination (control) or IVP. Fetal ultrasonographies were taken on gestation day 55 (D55) and fetal collections performed on D56 or D105 (gestation in cattle ~280 days). ART fetuses weighing ~97 percentile of the control weight were considered LOS. Ultrasonography results show that the product of six D55 measurements can be used to identify extreme cases of LOS. To determine whether maternal blood can be used to identify LOS, leukocyte mRNA from 23 females was sequenced. Unsupervised hierarchical clustering grouped the transcriptomes of the two females carrying the two largest LOS fetuses. Comparison of the leukocyte transcriptomes of these two females to the transcriptome of all other females identified several misregulated transcripts on gestation D55 and D105 with LOC783838 and PCDH1 being misregulated at both time-points. Together our data suggest that LOS is identifiable during pregnancy in cattle.

Dickkopf-1 (DKK1) is a secreted inhibitor of canonical WNT signaling expressed in the endometrium that can increase the competence of bovine blastocysts to establish pregnancy after transfer into recipients. The mechanism by which DKK1 regulates embryo function is not known. KREMEN1 and KREMEN2 are paralogs that function as DKK1 transmembrane receptors. Binding of DKK1 to KREMEN leads to internalization of the LRP5/6 WNT co-receptor and inhibition of β-catentin (CTNNB1)-dependent WNT signaling. Here we evaluated whether the bovine pre-implantation embryo expresses KREMEN1 and KREMEN2 and whether knockdown of KREMEN1 would alter embryonic development and accumulation of CTNNB1. In the first experiment, expression of KREMEN1 and KREMEN2 was determined in individual matured oocytes and embryos at the 2-cell, 3-4-cell, 5-8-cell, 9-16-cell, morula, and blastocyst stages of development (n=4) using RT-qPCR with GAPDH and YWHAZ as internal controls. Data were analysed by analysis of variance using GLM procedure of SAS (SAS Institute Inc.; stage, replicate in model), and Tukey test for multiple comparisons. Expression of both KREMEN1 (P&lt;0.0001) and KREMEN2 (P=0.003) varied between stages of development. There was a large decline in transcript abundance for KREMEN1 after the 9-16-cell stage. Expression (fold-change) relative to housekeeping genes was 0.29, 0.47, 0.27, 0.47, 0.34, 0.05, and 0.01 (standard error=0.08) for oocyte, 2-cell, 4-cell, 5-8 cell, 9-16-cell, morula, and blastocyst stages, respectively. Expression of KREMEN2 was low at early stages of development, increased at the 8-16 cell and morula stages (i.e. after embryonic genome activation), and then declined. Expression (fold-change) relative to housekeeping genes was 0.00, 0.04, 0.03, 0.05, 0.12, 0.15, and 0.01 (standard error=0.04) for oocyte, 2-cell, 4-cell, 5-8-cell, 9-16-cell, morula, and blastocyst stages, respectively. In the second experiment, we determined effects of a GapmeR antisense oligonucleotide directed against KREMEN1 on development to the blastocyst stage and amounts of immunoreactive CTNNB1 in the blastocyst. The experiment was performed in 3 replicates using 217 putative zygotes per treatment. The GapmeR treatment did not affect cleavage rate (78.5±0.02% for GapmeR vs. 71.7%±0.02% for vehicle; P=0.21), percent putative zygotes becoming blastocysts (23.4±0.02% vs. 20.0±0.02%; P=0.57), or the percent of cleaved embryos becoming blastocysts (30.6±0.03% vs. 28.8±0.03%; P=0.88). Amounts of immunoreactive CTNNB1 were determined in blastocysts collected at 168h after insemination (n=7/treatment). Treatment with GapmeR tended (P=0.08) to increase the fluorescence intensity of CTNNB1 (1761±68 vs. 1583±72 units). In summary, KREMEN1 is expressed before genome embryonic activation, whereas KREMEN2 is expressed in 8-16 cells and morula. Knocking down KREMEN1 does not compromise embryonic development, but preliminary studies indicate KREMEN1 can regulate CTNNB1 abundance. Support was provided by USDA-NIFA 2017-67015-26452.

Test the shipping incubator and make sure that it can remain powered for at least 24 hours while operating with the battery. We currently use TO 42 from WTA https://www.wtavet.com.br/?lang=en but others work well also. The temperature is set to 38.5°C Pi cture 2.j pg Pi cture 2.j pg 2 Start charging shipping incubator 1 day before shipping so that it is fully charged at the time tubes containing embryos are added to the incubator.