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The aim of the present study was to evaluate the effect of estradiol benzoate (EB) treatment on estradiol (E2) and progesterone (P4) in blood, and on estrogen (ER) and progesterone (PR) receptors in the uteri of mixed breed bitches. Injections of 10 µg EB/kg (Mesalin, Intervet) and 100 µg EB/kg (Oestradiolum benzoicum, Polfa) were given on day 3 and 5 after the first acceptance of males. Steroids were measured in samples collected from the vena cephalic using the immunofluorescent method. The average level of E2 was elevated to 190 and 260 pg/ml, respectively, and declined to control values within less than 12 days. An increase in P4 was observed only during the first 4 days. ER and PR were determined in uterine samples collected during hysterectomy performed on day 14 or 28 following the first EB injection. The radioligand assay was used for soluble (100.000 g cytosol) and structural (0.6 M NaCl extract) fractions. Typical binding characteristics for ER and PR were found in all analyzed samples. The amount of ER was unchanged over time in the untreated group, while PR significantly declined. The lower dose caused a 3-fold elevation of ER and PR (mainly soluble fractions) which on day 28 declined to control values with the exception of an uncompleted decline of soluble ER. The effect of higher doses on ER was only significant in its soluble fraction on day 14. However its influence on the PR (initially unchanged) occurred on day 28 and was elevated more than 10-times (mainly structural fraction). The results obtained indicate that uterine ER and PR responded differently to high and low doses of estrogen treatment.
In the present study we have evaluated the effect of estrogen contraception on ovarian and uterine morphology in healthy bitches. 30 crossbreed bitches with physiological estrous cycle (1-3 y; 10-14 kg) were divided into control (n = 6) and two experimental groups of 12 bitches treated twice with different doses of estradiol benzoate (EB). The first group received 10 µg EB/kg BW (Mesalin, Intervet), and the second 100 µg EB/kg BW (Oestradiolum benzoicum, Polfa), given i.m. on day 3 and 5 after the first acceptance of a male. The ovaries and uteri were collected 14 or 28 days from the first EB injection. The histopathological evaluation and histomorphometric measurements were performed using computer assisted analysis. There was no significant effect of estrogen treatment on ovarian activities at either dose. The uteri of bitches from the control group have shown progressive regression of all endometrial elements typical for the middle part of the luteal phase of the estrous cycle. The low dose EB treatment caused glandular hyperplasia characteristic for early stages of cystic formation. The higher dose resulted in total inhibition of cell proliferation, stimulation of glandular hyperplastic (complex) changes and increased thickness of the uterine wall. At the latter time the cystic changes within the glandular part may cause endometritis or even pyometra. The results obtained indicate the need for clinical follow up of bitches treated with estrogens.
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