Veterinary Research Communications, 27 Suppl. 1 (2003) 607–609 © 2003 Kluwer Academic Publishers. Printed in the Netherlands
Mare Embryonic Resorption and Homocysteine A. Giammarino1, D. Robbe1*, E. Dainese1, R. Minoia2 and R.L. Sciorsci2 1Department of Clinical Veterinary Science, University of T eramo; 2Department of Animal Production, University of Bari, Italy *Correspondence: Dipartimento di Scienze Cliniche Veterinarie, Sezione di Ostetricia, Facolta` di Medicina Veterinaria, V iale Crispi 212, 64100 T eramo, Italy E-mail:
[email protected] Keywords: embryonic resorption, homocysteine, mare
INTRODUCTION Early embryonic death in the mare is one of the main causes of subfertility in horses (Woods et al., 1987). Pregnancy loss in women has been correlated with hyperhomocysteinaemia (Steegers-Theunissem et al., 1997) and seems to be caused by an alteration of placental angiogenesis (Nagai et al., 2001). Hyperhomocysteinaemia is considered an important marker of obstetric and gynaecological conditions (Scholl and Johnson, 2000). The aim of the present study was to test the levels of homocysteine in mares suffering from embryonic death.
MATERIALS AND METHODS The experiment was carried out in a stud-farm in the Rome area, on 51 mares, Italian, French and American trotters aged 4 to 20 years, during the 2001 breeding season between the end of February and mid-June. The mares came from different Italian farms and were fed with forage and concentrate. Coprological examinations were performed on all animals to verify the degree of infestation by the most important intestinal parasites. The results were all negative. Physical examinations were performed on all mares. Ultrasound examinations, to confirm pregnancy, were carried out three times: on the 15th, 30th and 40th day after insemination, using a portable ultrasound unit (‘Dynamic Imaging’ with a 7.5 mHz linear probe). The control group of our experiment consisted of subjects in which embryonic reabsorption did not occur. Three blood samples were collected from each subject to determine homocysteine levels. Samples were always collected before morning meals. First sample (T .0): collected before insemination, when the follicle presented a diameter of 35 mm. Second sample and third sample (T .15 and T .40): collected upon diagnosis of pregnancy (day 15 and 40). The blood was drawn from the jugular vein and collected in sterilized vacutainers 607
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(Venoject). Samples were then refrigerated (4°C) and centrifuged (1200g×10 min) within 1 hour of drawing. The serum was stored in Eppendorf-like test-tubes and frozen at −20°C until analysis. To measure homocysteine levels, an ABBOT Diagnostic Division IMx System kit was used. The assay system is based on an immunofluorescence method using polarized light (FPIA). The homocysteine values obtained were subjected to statistical analysis with Student t test and ANOVA. Both statistical tests were performed using the SPSS program. RESULTS Of the 51 inseminated mares, 39 were diagnosed pregnant (76.47%), on the last ultrasound test on the 40th day, five were not pregnant (9.80%) and in seven (13.73%) reabsorption was detected on the first diagnosis. The homocysteine average value of pregnant mares and those which reabsorbed are reported in Figure 1. In the group of mares that reabsorbed the statistical comparison showed a significant difference between the T.0 and the T.40 ( p
