Somatic nuclear transfer (NT) in cattle is often complicated by fetal oversize (i.e., large offspring syndrome), hydrallantois, and placentomegaly in late gestation. The aims of this work were to obtain data on the placentome structure in NT-recipient cows with hydrallantois (NTH) and to relate these with fetal and placental weights to better understand the abnormalities observed in NTH pregnancies during the third trimester. Pregnant cows were slaughtered between Gestation Days 180 and 280. The fetuses were weighed, and the placentomes were numbered and weighed. Placentomes were examined by histologic and stereological techniques. Macroscopic data showed that placental overgrowth preceded fetal overgrowth, and the ratio of the fetal to the total placentome weight in the NTH group was lower than that in controls after Gestation Day 220. This suggests that placental overgrowth is due to placental default rather than due to fetal overgrowth, as shown also by stereological analysis showing primary deregulation of the growth of cotyledonary tissues. Observed alterations, such as thinning of the maternal epithelium within placentomes and increased trophoblastic surface, could be secondary adaptations. Thus, placental growth deregulations would be due to modifications of the expression of placental factors. Various examples of placental deficiency were observed, suggesting that some fetal abnormalities observed in NTH calves, such as enlarged heart, enlarged umbilical cord, and abdominal ascites, are consequences of placental dysfunction. Therefore, the condition described by the term "large offspring syndrome" might better be described by "large placenta syndrome," because this syndrome affects an average of 50% of late-gestation NT pregnancies. No conclusion can be drawn from this work on apparently normal pregnancies.

In contrast to endometritis, now diagnosed by cytological examination, the effect of endocervical inflammation on reproductive performance has been inadequately investigated. In this study, endocervical and endometrial cytological specimens were collected from 168 Holstein cows between 21 and 60 days in milk (DIM) to investigate the prevalence of endocervical inflammation and effect on days to conception. Statistical analyses were stratified based on DIM at examination (<35 vs. ≥35 DIM). Endocervical inflammation with ≥5% neutrophils before 35 DIM (disregarding the level of endometrial inflammation) was associated with decreased hazard of pregnancy within 300 DIM (adjusted hazard ratio, 0.4; 95% confidence interval: 0.3-0.8). A decrease in hazard of pregnancy was observed when >6% neutrophils were counted in endometrial smears (adjusted hazard ratio, 0.4; 95% confidence interval: 0.2-0.7). The study suggested an additive effect of combined endocervical and endometrial inflammation on the hazard of pregnancy within 300 DIM. Using the thresholds of 5% neutrophils for the cervix and 6% neutrophils for the uterus, 11% of the cows examined before 35 DIM presented cervicitis only, 13% were affected by endometritis only, and 32% suffered from both endometrial and endocervical inflammation. The presence (absence) of cervicitis was not indicative of the presence (absence) of endometritis. This study showed that in addition to uterine inflammation, endocervical inflammation in early lactation affects conception. Thus, the global evaluation of genital tract health may be more beneficial for reproductive performance than that of endometrial inflammation.

We analyzed the change in gene expression related to dam physiological status in day (D)18 embryos from growing heifers (GH), early lactating cows (ELC), and late lactating cows (LLC). Dam energy metabolism was characterized by measurement of circulating concentrations of insulin, glucose, IGF-1, nonesterified fatty acids, ␤-hydroxybutyrate, and urea before embryo flush. The metabolic parameters were related to differential gene expression in the extraembryonic tissues by correlation analysis. Embryo development estimated by measuring the length of the conceptuses and the proportion of expected D18 gastrulating stages was not different between the three groups of females. However, embryo metabolism was greatly affected by dam physiological status when we compared GH with ELC and GH with LLC but to a lesser extent when ELC was compared with LLC. Genes involved in glucose, pyruvate, and acetate utilization were upregulated in GH vs. ELC conceptuses (e.g., SLC2A1, PC, ACSS2, ACSS3). This was also true for the pentose pathway (PGD, TKT), which is involved in synthesis of ribose precursors of RNA and DNA. The pathways involved in lipid synthesis were also upregulated in GH vs. ELC. Despite similar morphological development, the molecular characteristics of the heifers' embryos were consistently different from those of the cows. Most of these differences were strongly related to metabolic/hormone patterns before insemination and during conceptus free-life. Many biosynthetic pathways appeared to be more active in heifer embryos than in cow embryos, and consequently they seemed to be healthier, and this may be more conducive to continue development. dairy cow; fertility; energy balance; energy metabolism; conceptus development; gene expression; correlations IN THE MODERN HOLSTEIN cow, calving rates are close to 55-60% in heifers and 35-40% in lactating cows (39,73). Most pregnancy losses occur prior to the period of maternal recognition and maintenance of the corpus luteum, i.e., before days 15-17 (D15-D17) after artificial insemination (AI), and are due to nonfertilization-early embryonic loss (NF-EEL, Refs. 22,35,73). NF-EEL can be estimated in field trials by measuring plasma or milk progesterone concentrations the day of AI and 21-24 days later. The frequency of NF-EEL has been reported to be between 25 and 45% in lactating Holstein cows (26,32,35,52), and it has increased from the 1980s (22).Age/parity, uterine health (nulliparous heifers vs. postpartum cows), and metabolic and endocrine environment are factors that can explain differences in reproductive efficiency. Maternal metabolism is known to influence fertility. Milk production, body condition score (BCS), postpartum BCS variations, negative energy balance (NEB), and quantity and quality of dietary protein supply are related to pregnancy rate (reviews Refs. 15,40,45,65,73,76). This effect might be explained partly by the clearance of progesterone in highproducing dairy cows (22) and by poor oocyte quality (43), which impair fertilization ability and early e...

BackgroundProgesterone (P4) may modulate oviductal functions to promote early embryo development in cattle. In addition to its nuclear receptor (PR), P4 may mediate its actions through P4 receptor membrane component 1 (PGRMC1) and its relative, PGRMC2. Two successive experiments were undertaken to characterise the expression of PR, PGRMC1 and PGRMC2 in the bovine oviduct during the post-ovulation period, and to relate their expression to the presence of an embryo, the proximity of the CL and to the region of the oviduct.MethodsIn the first experiment (Exp. I), whole oviduct sections were collected from Holstein cows at Day 1.5, Day 4 and Day 5 post-ovulation (n = 2 cows per stage). The expression of PR, PGRMC1 and PGRMC2 was studied in the ampulla and isthmus by RT-PCR, western-blot and immunohistochemistry. In Exp. II, oviduct epithelial cells were collected from cyclic and pregnant Charolais cows (n = 4 cows per status) at Day 3.5 post-ovulation and mRNA expression of PR, PGRMC1 and PGRMC2 was examined in the ampulla and isthmus by real-time quantitative PCR.ResultsIn Exp. I, PR, PGRMC1 and PGRMC2 were expressed in all oviduct samples. PGRMC1 was mainly localised in the luminal epithelium whereas PR and PGRMC2 were localised in the epithelium as well as in the muscle and stroma layers of the oviduct. The expression was primarily nuclear for PR, primarily cytoplasmic for PGRMC1 and both nuclear and cytoplasmic for PGRMC2. In Exp. II, mRNA levels for PR, PGRMC1 and PGRMC2 were not affected by either the pregnancy status or the side relative to the CL. However, the expression of PR and PGRMC2 varied significantly with the region of the oviduct: PR was more highly expressed in the isthmus whereas PGRMC2 was more highly expressed in the ampulla.ConclusionsThis is the first evidence of PGRMC2 expression in the bovine oviduct. Our findings suggest that P4 regulates the functions of the bovine oviduct in a region-specific manner and through both classical and non classical pathways during the post-ovulation period.