STUDIES ON ULTRASONOGRAPHY, PLASMA ENDOCRINE AND BIOCHEMICAL PROFILES DURING EARLY PREGNANCY IN MARES
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Date
2013
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AAU, Anand
Abstract
This research work entitled was carried out during breeding seasons on apparently healthy field (farmer) mares (n=32) selected at random from different areas of Saurastra region (March-August, 2011) and Thoroughbred mares (n=45) of private stud farm of North Gujarat (February-June, 2012). The mares of both the class were subjected to per rectal examinations, the Thoroughbred mares were also scanned by ultrasonography, for the pregnancy status and were divided into pregnant and non-pregnant groups. These mares (Field vs. Thoroughbred) were further distributed in three stages as per the pregnancy days, viz., Stage-I: pregnant mares between day 0 and 35 of gestation (4 vs. 12), Stage -11: comprises of pregnant mares between day 36 and 75 of gestation (7 vs. 12), Stage-III: comprises of pregnant mares between day 76 and 150 of gestation (11 vs. 11), and Stage-IV: non-pregnant mares (10 vs. 10). Blood plasma samples collected from above mares were analyzed for biochemical and hormonal profile. The mean plasma progesterone levels (ng/ml) of field and Thoroughbred pregnant mares were significantly higher in stage-III mares (20.34 ±2.13 vs. 29.32 ± 2.48) followed by stage-I (15.55 ± 3.54 vs. 17.29 ± 1.66) and stage II mares (13.51 ± 1.81 vs. 23.62 ± 2.31), than those in stage-IV non-pregnant mares (0.97 ± 0.15 vs. 1.68 ± 0.35) were observed in non-pregnant mares (stage-IV). The overall pooled mean progesterone profile of field and Thoroughbred mares showed an increasing trend from pregnancy stage-I to stage-III. Significantly higher mean progesterone level (ng/ml) was observed during stage-III (24.83 ± 1.87) of pregnancy than that at stage-I (16.86 ± 1.48), II (19.89 ± 1.94) and IV (1.33 ± 0.25). The mean progesterone levels (field mares vs. Thoroughbred mares) at stage-II (13.51 ± 1.81 vs. 23.62 ± 2.31) and stage-III (20.34 ±2.13 vs. 29.32 ± 2.48) were significantly (P < 0.05) higher in Thoroughbred mares than those of field mares. The mean plasma estradiol-17β levels (pg/ml) were significantly higher (P< 0.05) in field and Thoroughbred pregnant mares of stage-III (289.82 ± 73.05 vs. 240.91 ± 32.13) than those mares of stage-I (28.50 ± 4.52 vs. 91.92 ± 14.28) and stage-II (43.14 ± 11.90 vs. 86.00 ±7.15) and non-pregnant mares of stage-IV (23.80 ± 1.82 vs. 74.50 ± 6.37). There were non significant differences among stage-I, II and IV. The overall pooled values of estradiol-17β (pg/ml) of field and Thoroughbred mares were significantly higher during early pregnancy stage-III (265.36 ± 39.31) than those found during stage-I (76.06 ± 12.78), II (70.21 ± 7.80) and IV (49.15 ± 4.09). The mean estradiol-17β levels of Thoroughbred mares were significantly higher during stage-I (91.92 ± 14.28) and II (86.00 ± 7.15) and IV (74.50 ± 6.37) as compared to those of field mares. However, the estradiol-17β level of pregnancy stage-III was significantly higher in field mares than that of Thoroughbred mares (289.82 ± 73.05 vs. 240.91 ±32.13 pg/ml). The mean cholesterol levels of both the class of mares were significantly (P<0.05) higher during non-pregnancy stage-IV than the pregnancy stage-II, I and III. The overall mean total cholesterol levels among various stages of pregnancy did not differ significantly. The mean total cholesterol level (mg/dl) during stage-III of pregnancy was significantly higher in Thoroughbred mares (78.74 ±2.15) than those of field mares (64.15 ±3.22). The mean plasma triglycerides (mg/dl) level of pregnant field mares was significantly higher during stage-II (30.26 ± 3.65) followed by stage-III (29.33 ± 1.86) and I (27.64 ± 2.38) than in non-pregnant mares (20.56 ± 2.56). The mean triglycerides level (mg/dl) in Thoroughbred mares was, however significantly higher during stage-III (39.93 ± 2.69) than those of stage-II (26.24 ± 2.89), I (23.95 ± 3.21) of pregnancy and non-pregnancy stage-IV (25.55 ± 2.82). The overall pooled mean triglycerides concentration was significantly higher in pregnancy stage-III than those mares at stage-II, I and IV. The Thoroughbred mares had significantly higher mean plasma triglycerides (mg/dl) level than the field mares during pregnancy stage-III (39.93 ±2.69 vs. 29.33 ±1.86). The mean plasma total protein levels in field mares and also overall of two classes did not vary significantly among the stages of pregnancy and non-pregnancy. The mean total protein levels (g/dl) in Thoroughbred mares was, however significantly higher during stage-III (7.52 ± 0.76) than those of stage-II (6.98 ±0.15) and IV (6.77 ± 0.24). The mean plasma protein levels between field and Thoroughbred mares during various stages of pregnancy and non-pregnancy did not differ significantly. The mean plasma glucose (mg/dl) levels were significantly higher at stage-III of pregnancy in both field (101.40 ± 5.38) and Thoroughbred (102.18 ± 4.26) mares than other stages. The mean plasma glucose levels between field and Thoroughbred mares were identical at different stages of pregnancy and non-pregnancy. The mean plasma calcium (mg/dl) level in field mares was significantly lower at stage-I (10.65 ± 0.26) of pregnancy than those observed at stage-II (11.36 ± 0.32), III (11.98 ± 0.11) and IV (11.77 ± 0.17). The non-pregnant Thoroughbred farm mares showed significantly higher (12.73 ± 0.18 mg/dl) mean plasma calcium levels than those observed during all 3 pregnancy stages. The overall mean plasma calcium level during non-pregnancy (stage-IV) was significantly higher than those of pregnant mares in stage-III, II and I. The mean calcium levels between the field and Thoroughbred mares did not differ significantly at any of the stages of pregnancy and non-pregnancy. The mean plasma inorganic phosphorus (mg/dl) values were significantly higher during stage-III of pregnancy in both field (3.97 ± 0.06), Thoroughbred mares (3.76 ± 0.11) and hence the overall mean value (3.86 ± 0.06). The correlation studies revealed significant (P<0.05) and positive relationship of plasma cholesterol levels with total protein (r = 0.64) and phosphorus levels (r = 0.60) in Thoroughbred mares. The glucose levels observed during stage-l of pregnancy in Thoroughbred mares had significant and positive correlation with pregnancy days (r = 0.59). The triglyceride levels observed during gestational stage-II in Thoroughbred mares had significant (P<0.05) and positive correlations with total protein levels (r = 0.62). The plasma total protein levels had significant (P<0.05) and positive correlations with phosphorus levels (r = 0.62) during stage-II. The pregnancy days had significant (P<0.05) and positive correlations with triglycerides (r = 0.66), total protein (r = 0.81) and phosphorus levels (r = 0.92). In non-pregnant Thoroughbred mares, the glucose levels had significant (P<0.05) and positive correlation with phosphorus levels (r = 0.63). Pregnancy days had significant (P<0.05) and positive relationship with plasma glucose (r = 0.97) in field mares. The cholesterol levels observed during stage-Il of pregnancy in field mares had significant (P<0.05) and positive correlation with triglyceride levels (r = 0.79). The plasma calcium had significant (P<0.05) and positive correlafion with phosphorus levels (r = 0.82). The embryonic vesicle first became visible as a fluid filled structure, recognized as non-echogenic sphere of 9 mm and 17 mm on day 14 of pregnancy, respectively by the ultrasonography. The mean size of embryonic vesicles observed on day 14, 18-22, 28-30, 35, 45-48, 56 and 60-63 of pregnancy was 13.0 ± 4.0, 28.0 ± 0.82, 36.5 ±1.41, 52.0 ± 0.0, 69.0 ± 5.0,74.0 ± 0.0 and 92.2 ± 2.2 mm, respectively. With the routine examination of pregnant mares with ultrasonography, the embryo as well as the fetal heart beat was also detected for the first time on 28th day post covering. The mean size of embryo was 11.2 ± 0.95 mm during days 28-30 of gestation. The increase in the size of embryo was observed during gestational days 35, 45-56 and 60-63 and the corresponding size of the embryo during above gestational days was 22.0 ± 0.0, 29.67 ± 0.41 and 54.8 ± 0.63 mm, respectively. The study clearly indicated that the pregnancy can be diagnosed by ultrasonography as early as on day 14th of pregnancy. Similarly, hormonal profile determined in field and Thoroughbred mares showed that the higher (>13.5 ng/ml) and lower (< 3.7 ng/ml) mean progesterone levels were a good indicators of positive confirmation of pregnancy and non-pregnancy, respectively. Moreover, the determination of higher progesterone levels (> 8.2 ng/ml) and estradiol-17β (>120 pg/ml) during 14 to 150 days of gestation in an individual mare confirmed the early pregnancy, whereas lower progesterone (< 3.7 ng/ml) and estradiol-17β (<100 pg/ml) level confirmed the non-pregnancy. However, estradiol-17β value > 155 pg/ml between 76 to 150 days of pregnancy also offered the most reliable method of pregnancy diagnosis by radioimmunoassay. Hence, transrectal ultrasonography and plasma progesterone and estradiol-17β assay may be used alone or in combination for the confirmation of early pregnancy in mares.
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