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During the periovulatory period, the induction of prostaglandin G/H synthase-2 (PTGS2) expression in cumulus cells and associated prostaglandin E2 (PGE2) production are implicated in the terminal differentiation of the cumulus-oocyte complex. During the present study, the effects of the PTGS2/PGE2 pathway on the developmental competence of bovine oocytes were investigated using an in vitro model of maturation, fertilization, and early embryonic development. The specific inhibition of PTGS2 activity with NS-398 during in vitro maturation (IVM) significantly restricted mitogen-activated protein kinase (MAPK) activation in oocytes at the germinal vesicle breakdown stage and reduced both cumulus expansion and the maturation rate after 22 h of culture. In addition, significantly higher rates of abnormal meiotic spindle organization were observed after 26 h of culture. Periconceptional PTGS2 inhibition did not affect fertilization but significantly reduced the speed of embryo development. Embryo output rates were significantly decreased on Day 6 postfertilization but not on Day 7. However, total blastomere number was significantly lower in embryos obtained after PTGS2 inhibition. The addition of PGE2 to IVM and in vitro fertilization cultures containing NS-398 overrode oocyte maturation and early embryonic developmental defects. Protein and mRNA expression for the prostaglandin E receptor PTGER2 were found in oocytes, whereas the PTGER2, PTGER3, and PTGER4 subtypes were expressed in cumulus cells. This study is the first to report the involvement of PGE2 in oocyte MAPK activation during the maturation process. Taken together, these results indicate that PGE2-mediated interactions between somatic and germ cells during the periconceptional period promote both in vitro oocyte maturation and preimplantation embryonic development in cattle.

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Effects of postpartum energy restriction, parity and time after parturition on energy status (measured by glucose, insulin, non-esterified fatty acids (NEFAs) and beta-hydroxybutyrate), LH secretion and follicular growth were investigated in ten primiparous and nine multiparous suckled cows. Females were allocated by parity, body mass and body condition score at calving to diets supplying either 100% (CE, n = 10) or 70% (LE, n = 9) of energy requirements until day 70 postpartum. Metabolic parameters were measured every week from calving to day 70 postpartum. Blood samples were collected at intervals of 15 min for 10 h on day 30 and day 50 after parturition for LH measurement. Ovaries were examined between days 20 and 30 and days 40 and 50 postpartum by ultrasonography. Energy supply affected mean plasma concentrations of glucose (CE: 0.64 +/- 0.01 g l-1 versus LE: 0.61 +/- 0.01 g l-1; P < 0.05) and NEFA (CE: 168 +/- 17 mu eq l-1 versus LE: 309 +/- 18 mu eq l-1; P < 0.01) but by day 70 postpartum, glucose and NEFA concentrations were not significantly different between the two groups. LH pulse amplitude and frequency were not affected by energy supply (P > 0.10). However, at day 30 postpartum, LH pulse frequency was negatively correlated with plasma concentration of NEFA (r = -0.61; P < 0.01). Cows fed diets supplying 100% of energy requirements had more large follicles than did cows fed low energy diets (CE: 0.82 +/- 0.05 versus LE: 0.31 +/- 0.05; P < 0.05). The size of the largest follicle was greater in CE cows than in LE cows (CE: 10.2 +/- 0.1 mm versus LE: 8.7 +/- 0.2 mm; P < 0.05). Between 40 and 50 days postpartum, the size of the largest follicle was negatively correlated with NEFA concentration (r = -0.5; P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Prostaglandin E 2 (PGE 2 ) and progesterone appear to be critical mediators of cumulus expansion and the resumption of oocyte meiosis. The aim of this study was to identify the types of prostaglandin E synthase (PTGES) expressed in the bovine cumulus-oocyte complex (COC), to characterize their temporal expression during the periconceptional interval using an in vitro model of maturation (IVM) and fertilization (IVF), and to compare their expression with the level of steroidogenic gene expression. Real-time RT-PCR analysis revealed that enzymes related to the PGE 2 biosynthesis pathway were mainly expressed during IVM. Transcripts encoding PTGES1-3 were detected in bovine COCs. Only the expression of PTGES1 significantly increased during IVM whereas that of PTGES2 and PTGES3 remained unchanged. The induction of PTGES1 expression paralleled the induction of prostaglandin G/H synthase-2 (PTGS2) expression and the amounts of PGE 2 secreted by maturing COCs. Concomitantly, cholesterol side chain cleavage cytochrome P450 expression was significantly upregulated in maturing COCs and the high level of expression persisted in fertilized COCs. The expression of the StAR protein remained constant during IVM and then decreased significantly during IVF. Expression of the progesterone catabolic-related enzyme, 20a-hydroxysteroid dehydrogenase significantly decreased throughout the periconceptional interval. This was associated with a rising level of progesterone released by COCs in the culture media. In conclusion, our results suggest that the periconceptional differentiation of the bovine COC includes the transient induction of PGE 2 biosynthetic activity via the PTGS2/PTGES1 pathway during the maturation period and the increasing ability to produce progesterone from the immature to the fertilized stages. Reproduction (2008) 135 593-603

Bartonella spp. are small hemotropic bacteria infecting mammals. Four Bartonella species have been recently described in cattle and wild ruminants. To date, the biology and possible pathogenic role of Bartonella species isolated from ruminants are poorly understood. Therefore, a dairy herd of 448 cows and heifers was surveyed in order to establish the prevalence of Bartonella bovis and B. chomelii infections, the level of bacteremia, and the relationship between bacteremia and age or pregnancy status. The putative impact of Bartonella infection on production performance (individual milk cell count, milk yield) and reproductive status (success of artificial insemination [AI], placental retention, embryonic death, and abortion) was also assessed. The overall mean prevalence of B. bovis bacteremia was 59%, with the highest prevalence in heifers (92.5%). No B. chomelii was isolated, and 95% (114/120) of the B. bovis strains isolated and tested by PCR-restriction fragment length polymorphism belonged to type I. The level of bacteremia was higher in pregnant cows than in nonpregnant cows (P ‫؍‬ 0.05), and the level of bacteremia rose during the last two-thirds of gestation (P < 0.001). There was no correlation between bacteremia and milk yield, individual milk cell count, success of first AI, interval between two calvings, or incidence of abortion and embryonic death. The interval from calving to first AI was shorter and the incidence of placental retention was lower in bacteremic animals than in nonbacteremic ones (P ‫؍‬ 0.03 and P ‫؍‬ 0.01, respectively).

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...

The aim of our study was to test whether a reduction in dietary intake could improve in vitro embryo production in superovulated overfed dairy heifers. Cumulus-oocyte complexes of 16 Prim' Holstein heifers (14 6 1 months old) were collected by ovum pick-up (OPU), every 2 weeks following superovulation treatment with 250 mg FSH, before being matured and fertilized in vitro. Embryos were cultured in Synthetic Oviduct Fluid medium for 7 days. Heifers were fed with hay, soybean meal, barley, minerals and vitamins. From OPU 1 to 4 (period 1), all heifers received individually for 8 weeks a diet formulated for a 1000 g/day live-weight gain. From OPU 5 to 8 (period 2), the heifers were allocated to one of two diets (1000 or 600 g/day) for 8 weeks. Heifers' growth rates were monitored and plasma concentrations of metabolites, metabolic and reproductive hormones were measured each week. Mean live-weight gain observed during period 1 was 950 6 80 g/day (n 5 16). In period 2 it was 730 6 70 (n 5 8) and 1300 6 70 g/day (n 5 8) for restricted and overfed groups respectively. When comparing period 1 and period 2 within groups, significant differences were found. In the restricted group, a higher blastocyst rate, greater proportions of grade 1-3 and grade 1 embryos, associated with higher estradiol at OPU and lower glucose and b-hydroxybutyrate, were observed in period 2 compared with period 1. Moreover, after 6 weeks of dietary restriction (OPU 7), numbers of day 7 total embryos, blastocysts and grade 1-3 embryos had significantly increased. On the contrary, in the overfed group, we observed more <8 mm follicles 2 days before superovulation treatment, higher insulin and IGF-I and lower nonesterified fatty acids in period 2 compared with period 1 (no significant difference between periods for embryo production). After 6 weeks of 1300 g/day live-weight gain (OPU 7), embryo production began to decrease. Whatever the group, oocyte collection did not differ between period 1 and 2. These data suggest that following a period of overfeeding, a short-term dietary intake restriction (6 weeks in our study) may improve blastocyst production and embryo quality when they are low. However, nutritional recommendations aiming to optimize both follicular growth and embryonic development may be different.