Sequential Hormonal Changes in the Postpartum Dairy Cow

Peripheral plasma levels of 15-keto-13,14-dihydro-PGF2α, progesterone, Cortisol, LH and prolactin were studied in 6 primiparous postpartum dairy cows. The cows were followed by hormone measurements and clinical examinations from parturition until pregnancy was established. Blood was collected 3 times per day. The cervix, uterus and ovaries were examined by rectal palpation at 6–10 days intervals. The cows were observed for signs of oestrus twice daily and were additionally teased with a bull to provoke standing heat. Four cows had a normal parturition and dropped their fetal membranes shortly afterwards. (NR group). The remaining 2 retained their fetal membranes for more than 24 h following parturition (RFM group). One out of 6 cows showed standing oestrus at the first ovulation, 4 animals were in oestrus at the second ovulation and all cows showed signs of oestrus at the third ovulation. Although the length of the first luteal phase varied from 9 to 22 days a corpus luteum was in all cases palpated. The secretion of progesterone during the first luteal phase was terminated by a PGF2α release. A significant difference in 15-keto-13,14-dihydro-PGF2α levels between the 2 groups was found on days 0–4 (2.39 vs 6.87 nmol/1 at Ρ < 0.06). Postpartum prostaglandin F2α release as reflected by the level of 15-keto-13,14-dihydro-PGF2α lasted shorter in the NR group than in the RFM group (15–17 vs 21 days). Significant positive correlations between 15-keto-13,14-dihydro-PGF2α and Cortisol as well as between prolactin and Cortisol during the first 24 days postpartum were noted only in cows having normal parturition. The most pronounced daily prolactin variations occurred during the second luteal phase (NR group), when a significant difference between the times 8.00, 12.00 and 15.00 was recorded (14.7, 31.5 and 19.7 μg/l, respectively). Moreover, a partial negative correlation between log value of prolactin and arithmetical value of LH was found in these cows only during the first luteal phase after parturition.     

Distribution of sulphate and iduronic acid residues in heparin and heparan sulphate

1. A method was developed for determination of the uronic acid composition of heparin-like glycosaminoglycans. Polymers or oligosaccharides are degraded to monosaccharides by a combination of acid hydrolysis and deamination with HNO₂. The resulting uronic acid monosaccharides (accounting for about 70% of the uronic acid contents of the starting materials) are isolated and converted into the corresponding aldono-1,4-lactones, which are separated by g.l.c. The calculated ratios of glucuronic acid/iduronic acid are reproducible within 5%. 2. Samples of heparin from pig intestinal mucosa (molar ratio of sulphate/disaccharide unit, 2.40) and heparan sulphate from human aorta (sulphate/disaccharide ratio, 0.46) were subjected to uronic acid analysis. l-Iduronic acid constituted 77% and 19% respectively of the total uronic acid contents. 3. The correlation between the contents of sulphate and iduronic acid indicated by this finding also applied to the fractionated deamination products of the two polymers. The sulphated fragments varied in size from disaccharide to octasaccharide (or larger) and showed sulphate/disaccharide molar ratios in the range of 0.05–2.0. The proportion of iduronic acid increased with increasing ester sulphate contents of the oligosaccharides. 4. Previous studies on the biosynthesis of heparin in a cell-free system have shown that l-iduronic acid residues are formed by C-5 epimerization of d-glucuronic acid units at the polymer level; the process requires concomitant sulphation of the polymer. The results obtained in the present structural study conform to these findings, and suggest further that similar mechanisms may operate in the biosynthesis of heparan sulphate. The epimerization reaction appears to be linked to the sulphation of hydroxyl groups but does not seem to require sulphation of the target uronic acid residues. The significance of sulphamino groups in relation to the formation of iduronic acid is unknown.     

Progesterone determinations and clinical examinations of reproductive organs in purebred and crossbred female Zebu cattle

Five experiments were undertaken to investigate variation in progesterone concentrations as related to the collection and handling of samples of milk and blood, to determine reference values for progesterone and to evaluate clinical findings in relation to progesterone data from pure- and crossbred Zebu cattle reared in the Peruvian tropics. Whole-milk progesterone concentrations obtained from 12 Holstein x Nellore pregnant cows at hourly intervals over a 24-h period were highest immediately after milking; this peak was followed by a sharp drop over the next 3 h. Milk-fat content from 28 Brown Swiss x Nellore pregnant cows increased from 2.4% before milking to 6.7% afterwards (P < 0.05), whereas progesterone concentrations in whole milk increased from 18.4 to 59.5 nmol/1 (P < 0.05). Progesterone concentrations in fat-free milk were stable, with the exception of the fore-milk sample, which was lower than subsequent samples collected during milking (P < 0.05). Blood samples collected from 23 purebred, pregnant Nellore cows, were divided into four aliquots, and plasma and serum were harvested periodically over the next 120 h of storage at +4 degrees C, or in the shade at ambient air temperatures. The results indicate that blood samples can be stored unseparated at both temperatures studied for up to 3 h without severe loss of progesterone. Milk samples collected at different intervals during the luteal phase of the estrous cycle and during early and mid-pregnancy from crossbred cows showed no significant differences in progesterone concentrations between Days 9 to 13 of the cycle and Days 9 to 13 of gestation. Progesterone levels during advanced gestation were higher (P < 0.05). Out of 2,607 clinical examinations, the level of agreement between palpatory findings and progesterone determinations was 95.6 and 81.9% in diagnosing non-luteal and luteal structures, respectively. Significant differences in the level of agreement between palpators were found (P < 0.01). It is concluded that milk samples, preferably composite milk or strippings, must be consistently collected at the same stage of milking, and that centrifugation of blood samples should be done as soon as possible and not later than 3 h after collection.     

Endocrine profiles and embryo quality in the PMSG-PGF2alpha treated cow

Plasma progesterone and LH concentrations around estrus were determined for both PMSG treated (experimental animals) and non-treated (control animals) dairy cows and heifers of the Holstein Friesian and Jersey breeds, and these hormone profiles were related to the embryo quality. Most experimental animals experienced an increase in progesterone concentrations following PMSG treatment and an abrupt decrease to values below 3 nmol/l after PG injection. The mean (+/-SE) intervals from prostaglandin treatment to estrus were 46.9+/-1.8 h and 64.5+/-4.8 h for experimental and control animals, respectively. At the onset of heat the progesterone concentration in experimental animals with optimal embryo quality (group I) was significantly lower (p<0.01) than in experimental animals which yielded unfertilized eggs (group II) (1.2+/-0.1 versus 3.9+/-0.8 nmol/l) and significantly higher than the level in the control group (0.6+/-0.1 nmol/l). Following estrus the progesterone profiles in all 3 groups were studied and the length of the superovulatory cycle was measured to 26.0+/-4.8 days. The preovulatory LH surge occurred sooner after prostaglandin injection in experimental (41 h) than in control animals (65 h). The LH surge in group I occurred within a narrow range and reached a higher average level than group II (24.2+/-2.2 ng/ml and 16.3+/-3.7 ng/ml, respectively). The control group attained an even higher LH surge (31.8+/-8.8 ng/ml) than did the experimental animals. The data presented in this experiment indicate that plasma levels of progesterone and LH in PMSG-PGF(2)alpha treated animals are related to embryo or egg quality.     

Post-partum release of prostaglandin F(2alpha) and uterine involution in the cow

Peripheral plasma levels of the main blood plasma metabolite of PGF(2alpha) (15-keto-13,14-dihydro-PGF(2alpha)) and progesterone were investigated during the immediate, post-partum period in 59 normally calving cows. Uterine involution was monitored by weekly rectal palpations. The levels of the prostaglandin metabolite were high at parturition and remained thereafter elevated for periods varying up to 7-23 days. Uterine involution was completed during periods ranging from 16-53 days. According to the clinical findings, the animals were divided into three groups. Group A comprises 46 animals which had an uncomplicated, puerperal period. A significant (p<0.001) correlation between the duration of elevated prostaglandin levels and the time for completed uterine involution (Y=29.6 - 1.3 (X - 13.5)) was found for these animals. Group B animals (n=8) had periods of varying length with uterine discharge during the first 30 days post-partum. When compared to group A animals, the animals in group B had comparatively longer periods of prostaglandin release and also longer periods for completion of uterine involution. Group C animals (n=5) at times had palpable, thin-walled, cystlike structures in the ovaries during the first 30 days post-partum. In this group of animals, the periods of high prostaglandin levels, as well as for the completion of uterine involution, were similar to those for the animals in group A. Progesterone levels remained low during the immediate post-partum period and in no case were elevated levels found until the prostaglandin release had ceased.     

Plasma Cortisol in the Horse,Diurnal Rhythm and Effects of Exogenous Acth

Peripheral blood plasma Cortisol concentration and its diurnal variation was measured in 4 horses. Mean concentration of Cortisol during 24 hrs. was 42 ng/ml (s ± 20 ng/ml). Peak values occurred at 6 a.m. and the lowest values were observed at about 6 p.m. (mean 65 ng/ml and 20 ng/ml, respectively). Long-acting ACTH at a dose of 150 i.u. was given by intramuscular injection to the 4 horses. Peak Cortisol concentrations markedly exceeding the prestimulation level were obtained between 2 and 4 hrs. after injection. During the immediate 24 hrs. after these peaks, the mean Cortisol level was markedly lower and the cyclic variation out of phase with the basal diurnal pattern. After a gradual adjustment during the second postinjection day, no differences could be seen between the 2 patterns on day 3 after injection.