Dynamics of changes in the serum levels of cholesterol, thyroid and ovarian hormones in the postpartum period in ewes]

Postparturient anoestry, spontaneously changing over to seasonal anoestry, represents an important reserve of full utilization of the reproductive potential of sheep. In spite of the fact that the length of gestation in sheep only amounts to 148 days (+/- 5 days), inability of most sheep to enter the cycle during the spring season acts as a factor limiting the number of lambings--and at the same time production of lambs--to one lambing per year. In order to obtain more detailed knowledge of hormonal conditions in sheep puerpery, the presented work was directed at the study of dynamic changes in concentrations of thyroxine (T4), triiodothyronine (T3), 17 beta-estradiol (E2), progesterone (P4) and cholesterol (Chol) in the blood serum of ewes and at their mutual correlative dependences between the 36th hour and the 51st postparturient day. The observation was carried out in nine nursing ewes of the Slovak Merino breed, with average weight 40-50 kg, lambed in January and February. Blood was sampled by means of jugular vein puncture 14 days (-14th day) before parturition (a. p.), up to 36 h after parturition (p. p.) and on day 4, 7, 14, 17, 21, 25, 34, 42 and 51 p. p. In the period from the 36th h to day 21 p. p., concentrations of T4 (Tab. I, Fig. 1) showed a decreasing tendency compared to the starting -14th day (69.55 +/- +/- 0.12 nmol.-1) with the exception of a temporary increase on the 7th day p. p. The lowest values were recorded on days 4 and 21 p. p. (45.66 +/- 21.61 nmol.l-1, P less than 0.05 and 54.89 +/- 11.06 nmol.l-1, P less than 0.05, resp.). An increase of these values to the starting level was observed between days 25 and 51 p. p. Compared to the values on -14th day (0.76 +/- 0.22 nmol.l-1), a significant increase of T3 concentrations (Tab. I, Fig. 2) was recorded at the 36th h and on days 4 and 7 p. p. with the highest values (1.48 +/- +/- 0.34 nmol.l-1, P less than 0.001) at 36 h p. p. After the temporary decrease between days 14 and 21 p. p. its concentrations showed a constantly rising tendency statistically significant on days 34, 42 and 51 p. p., compared to the -14th day (P less than 0.05). Concentrations of E2 (Tab. I, Fig. 3) reached the values of 0.44 +/- 0.41 nmol.l-1 on the -14th day.(ABSTRACT TRUNCATED AT 400 WORDS)     

Bacteriological and cytological findings during the late puerperal period after two different treatments of retained placenta followed by acute puerperal metritis

The aim of the study was to compare the effect of two acute puerperal metritis (APM) treatment protocols on uterine condition during the late puerperal period (5^th to 7^th week). Late gestation healthy cows (n = 21) were divided randomly in three equal groups. Parturitions were induced. Treatments of APM were started on the third day postpartum (PP). Group A was treated with an oxytocin analogue carbetocin for three days and intrauterine administration of cephapirin between days 15 and 17. Group B was given intramuscular injection of ceftiofur for five days followed by two injections of prostaglandin F_2α, at an interval of 12 h, on the eighth day PP. Group C served as the control group with no treatment. Body temperature was recorded daily for 14 days PP. Uterine biopsies for bacteriology, and uterobrush samples for cytology, were taken once a week from the 5^th to 7^th week postpartum. No differences were found in body temperature on day 14 PP, presence of bacteriological infections and disappearance of uterine inflammatory signs diagnosed by cytological examination between experimental groups.     

Morphologic changes in the ovary and histologic changes in the tertiary ovarian follicles in ewes after prolonged irradiation during anestrus

The changes in volume, weight and the histomorphological changes of the tertiary follicles of ewes were studied after protracted irradiation with 4.8 Gy in the anoestrous period by the morphometric and qualitative histological methods. The trial was performed in May with 21 ewes of the Slovak Merino breed, divided into three groups. The first group (five ewes) was control. The second and third groups (each containing eight ewes) were exposed to gamma-rays for five days, the total dose being 4.8 Gy. Within ten days after the treatment, all the irradiated and control ewes were given Ampicillin Spofa per os at a dose of 250 mg per head/day and Roboran Spofa at a dose of 10 g per head/day. The animals were killed by bleeding on the fifth day of irradiation and on the tenth day after the end of the treatment. After killing, the volume and weight of the ovaries were determined and a common histological method was used to cut these ovaries into 7 microns slices in series 70 microns apart. The slices were stained with haematoxylin-eosine and were evaluated by means of light microscopy. After irradiation the weight of the ovaries was found to decrease significantly; however, ovary volume remained unchanged. The atretic and non-atretic tertiary follicles were subjected to qualitative histological differentiation after Marion et al. (1968) and the number of non-atretic follicles was found to have decreased significantly in the irradiated ewes. The late type of atresia contributes most significantly to an increase in the proportion of atretic tertiary follicles. The administration of vitamins after irradiation reduced the occurrence of atretic changes.     

Characterization of gonadotropic and ovarian steroid hormones during the periovulatory period in high ovulating select and control line gilts

Cyclic gilts from Control (C, randomly selected, n = 11) and Relax Select (RS, nine generations of selection for increased ovulation rate followed by seven generations of relaxed or random selection, n = 9) lines of the University of Nebraska Gene Pool population (derived from 14 different breeds) were utilized to characterize differences in gonadotropic and ovarian steroid hormones during preovulatory and postovulatory phases of the estrous cycle. Blood samples were collected during four periods (0500, 1100, 1700 and 2300) daily beginning 2 d prior to anticipated estrus (d -2, d 18 of a 20-d estrous cycle), and continuing through d 4 postestrus (d 0 = 1st of standing estrus). Sampling within a period consisted of five blood samples at 15-min intervals. All plasma samples were analyzed for concentrations of follicle stimulating hormone (FSH) and luteinizing hormone (LH). Neither mean LH nor peak concentration of LH during the preovulatory surge differed between genetic lines (P greater than .10). Concentrations of FSH increased faster (line X period, P less than .05) and tended (P less than .1) to peak at a higher concentration in RS (.88 ng/ml) than in C (.54 ng/ml) gilts (P less than .05) during the 12 h preceding the FSH and LH preovulatory peaks. The second FSH surge began approximately 24 h after the preovulatory FSH peak. Peak FSH concentrations were observed at 42 h in both lines (1.46 vs 1.74 ng/ml for C and RS gilts, respectively). The higher FSH concentration in RS gilts established during the preovulatory surge was maintained through the second FSH surge (P less than .01). No line differences were detected in plasma concentrations of estradiol-17 beta and progesterone.     

Endocrine and ovarian changes in response to the ram effect in medroxyprogesterone acetate-primed Corriedale ewes during the breeding and nonbreeding season

Two experiments were performed to determine the endocrine and ovarian changes in medroxyprogesterone acetate (MAP)-primed ewes after ram introduction. Experiment 1 was performed during the mid-breeding season with 71 ewes primed with an intravaginal MAP sponge for 12 days. While the control (C) ewes (n = 35) were in permanent contact with rams, the ram effect (RE) ewes (n = 36) were isolated for 34 days prior to contact with rams. At sponge withdrawal, all ewes were joined with eight sexually experienced marking Corriedale rams and estrus was recorded over the next 4 days. The ovaries were observed by laparoscopy 4-6 days after estrus. Four weeks later, pregnancy was determined by transrectal ultrasonography. In eight ewes from each group, ovaries were ultrasonographically scanned; FSH, LH, and estradiol-17beta were measured every 12 hours until ovulation or 96 hours after estrus. The response to the rams was not affected by the fact that ewes had been kept or not in close contact with males before teasing. No differences were found in FSH, LH, estradiol-17beta concentrations, growth of the ovulatory follicle, onset of estrus, ovulation rate, or pregnancy rate. Experiment 2 was performed with 14 ewes during the nonbreeding season. Ewes were isolated from rams for 1 month, and received a 6-day MAP priming. Ovaries were ultrasonographically scanned every 12 hours, and FSH, LH, estradiol-17beta, and progesterone were measured. Ewes that ovulated and came into estrus had higher FSH and estradiol-17beta levels before introduction of the rams than did ewes that had a silent ovulation. The endocrine pattern of the induced follicular phase of ewes that came into estrus was more similar to a normal follicular phase, than in ewes that had a silent ovulation. The follicle that finally ovulated tended to emerge earlier and in a more synchronized fashion in those ewes that did come into estrus. All ewes that ovulated had an LH surge and reached higher maximum FSH levels than ewes that did not ovulate, none of which had an LH surge. We conclude that (a) the effect of ram introduction in cyclic ewes treated with MAP may vary depending on the time of the breeding season at which teasing is performed; (b) patterns of FSH, and estradiol-17beta concentrations, as indicators of activity of the reproductive axis, may be used to classify depth of anestrus; and (c) the endocrine pattern of the induced follicular phase, which is related to the depth of anestrus, may be reflected in the behavioral responses to MAP priming and the ram effect.     

Acute-phase proteins,cortisol and haematological parameters in ewes during the periparturient period

Numerous subclinical diseases in sheep occur in the periparturient period and involve inflammatory processes; therefore, determining markers, such as acute-phase proteins (APPs), can allow an early diagnosis. Therefore, the objective of the present study was to assess changes in the plasma concentration of APPs and cortisol in clinically healthy ewes in the periparturient period for use in future studies. At the same time, haematological parameters were monitored. We showed that plasma APPs and cortisol concentrations were significantly higher in pregnant ewes than before insemination. A gradual increase in the SAA concentration was observed from the 14th day before to the day of parturition, while Hp was reduced from 2 weeks before up to 2 weeks after delivery. A significant increase in the Fb concentration was detected from the 14th day before to the 1st week after delivery. The cortisol concentration did not undergo significant changes in the periparturient period. We found an increase in the SAA and Fb concentrations and decrease in Hp in the periparturient period. The direction of the change in APPs of healthy ewes in the current study may be related to their distinct regulatory mechanisms during pregnancy. The APPs are usually altered during infection, inflammation, neoplasia, stress and trauma; therefore, knowing their reference values could help lead to an early diagnosis of subclinical forms of some diseases and pregnancy complications in ewes. The haematological analysis showed that ewes in late pregnancy and postpartum compared to dry period were under metabolic stress related to pregnancy and lactation.     

Dynamics of protozoan counts in the rumen of calves during a period of milk feeding]

Protozoan counts were followed in the rumen of calves in the period of milk diet. In the third week of calves' age the total counts of protozoans in 1 ml of rumen contents made 1.48 +/- 0.91 (log 10 +/- SEM). These counts were increasing gradually to reach in the seventh week of age the value of 5.54 +/- 0.19 (log 10 +/- SEM) in 1 ml of rumen contents. An increase in the protozoan counts in the seventh week of age was significant in comparison with the preceding four weeks. Total protozoan counts were converted also per 1 g of rumen content dry matter.     

Dynamics of basic hematologic values in ewes during the course of a year

Basal hematological values were studied in ten primiparous ewes of Merino breed at 15 time intervals in the period from birth to the last decade of the next gravidity. The mean erythrocyte counts ranged from 6.5 to 10.3 T . l-1, hemoglobin content from 101.3 to 121.3 g per l, hematocrit value from 0.32 to 0.37 1 per l, mean corpuscular hemoglobin concentration from 29.97 to 34.22 g per dl, mean corpuscular hemoglobin from 11.17 to 16.61 pg, corpuscular volume from 35.03 to 55.37 fl. The mean leucocyte counts were from 4.98 to 9.93 G . l-1, with 25.8 to 60.5% neutrophil granulocytes with segmented nuclei, 0.1 to 2.6% neutrophil granulocytes with rodlike nuclei, 0.8 to 8.7% eosinophil granulocytes, 0.0 to 0.33% basophil granulocytes, 30.3 to 71.2% lymphocytes and 0.4 to 4.5% monocytes. The quality of red blood component was not negatively influenced by a high degree of gravidity. In the time of parturition, neutrophil granulocytes highly dominated over the other types of white blood cells; the counts normalized since the seventh day post partum. Although statistically significant differences were calculated for some time intervals, the values were within the physiological limits.     

The behavior of lactogenic and steroid hormones in the blood of Awassi ewes in Syria during lactation

The concentrations of lactogenic hormones (prolactin--PRL, growth hormone--GH, insulin-like growth factor I--IGF-I) and steroid hormones (progesterone--PG, estradiol--E2) were determined by RIA in the blood plasma of 8 lactating ewes (3 with twins) and compared with that of 3 non-lactating cycling ewes of the same age. All animals live in a flock of 30 Awassi sheep under identical conditions in Syria. The lactation period (192 +/- 25 days) is divided to a suckling period (until day 63) and a hand milking period (2 fold daily) with different levels of milk production (near 2 litres/day in the former, decreasing from 0.8 to 0.1 litre/day in the latter). During the suckling period the concentrations of steroid hormones in the blood plasma are lower, these of lactogenic hormones are higher of lactating than of non-lactating ewes. During the milking period only the concentrations of E2, PRL and IGF-I in the blood plasma of lactating animals are lower than in non-lactating ewes. But, during this time more and more seasonal influences (increasing day length and temperature) overwhelm the influence of lactation on the hormone secretion. Suckling of twins is accompanied with higher levels of E2, PRL and IGF-I in the blood plasma of their mothers than suckling of single lambs. During the lactation period a strong correlation exists between milk yield and the level of GH (r = 0.85) and IGF-I (r = 0.71), a smaller correlations to the level of E2 (r = 0.49) in the blood plasma of the lactating ewes. During the suckling period the milk yield is influenced positively by PRL (r = 0.77) and GH (r = 0.68), but negatively by PG (r = -0.76). During the milking period the milk yield is determined extensively by the level of IGF-I (r = 0.89) in the blood plasma. The concentrations of the analyzed hormones in the blood plasma correlate always positively together.     

Feed intake of ewes in the perinatal period

8 individual feeding experiments served the ascertainment of the consumption performance of ewes in the perinatal phase. In 6 experiments with a total of 176 fertility oriented ewes the daily intake of pelleted straw-concentrate mixtures between the 4th week ante partum and the 6th week post partum was determined. On average the ewes consumed 1701 g dry matter in the last phase of gestation and 2271 g DM during lactation per animal and day. Intense grinding and NaOH-treatment increased feed intake; high straw content, limited duration of feeding and NH3-treatment decreased it. A higher number of lambs had, in the last phase of gestation, a decreasing and during lactation an increasing effect on consumption. Maize silage in the last phase of gestation resulted in very low feed intake. In contra to this, rations of alfalfa hay and maize silage were consumed well during lactation. There were no significant differences due to species or age with regard to consumption performance.     

Comparative analysis between endometrial proteomes of pregnant and non-pregnant ewes during the peri-implantation period

Background:Early pregnancy failure has a profound impact on both human reproductive health and animal production.2/3 pregnancy failures occur during the peri-implantation period;however,the underlying mechanism(s)remains unclear.Well-organized modification of the endometrium to a receptive state is critical to establish pregnancy.Aberrant endometrial modification during implantation is thought to be largely responsible for early pregnancy loss.Result:In this study,using well-managed recipient ewes that received embryo transfer as model,we compared the endometrial proteome between pregnant and non-pregnant ewes during implantation period.After embryo transfer,recipients were assigned as pregnant or non-pregnant ewes according to the presence or absence of an elongated conceptus at Day 17 of pregnancy.By comparing the endometrial proteomic profiles between pregnant and non-pregnant ewes,we identified 94 and 257 differentially expressed proteins(DEPs) in the endometrial caruncular and intercaruncular areas,respectively.Functional analysis showed that the DEPs were mainly associated with immune response,nutrient transport and utilization,as well as proteasome-mediated proteolysis.Conclusion:These analysis imply that dysfunction of these biological processes or pathways of DEP in the endometrium is highly associated with early pregnancy loss.In addition,many proteins that are essential for the establishment of pregnancy showed dysregulation in the endometrium of non-pregnant ewes.These proteins,as potential candidates,may contribute to early pregnancy loss.     

Comparative analysis between endometrial proteomes of pregnant and non-pregnant ewes during the peri-implantation period

Background

Early pregnancy failure has a profound impact on both human reproductive health and animal production. 2/3 pregnancy failures occur during the peri-implantation period; however, the underlying mechanism(s) remains unclear. Well-organized modification of the endometrium to a receptive state is critical to establish pregnancy. Aberrant endometrial modification during implantation is thought to be largely responsible for early pregnancy loss.

Result

In this study, using well-managed recipient ewes that received embryo transfer as model, we compared the endometrial proteome between pregnant and non-pregnant ewes during implantation period. After embryo transfer, recipients were assigned as pregnant or non-pregnant ewes according to the presence or absence of an elongated conceptus at Day 17 of pregnancy. By comparing the endometrial proteomic profiles between pregnant and non-pregnant ewes, we identified 94 and 257 differentially expressed proteins (DEPs) in the endometrial caruncular and intercaruncular areas, respectively. Functional analysis showed that the DEPs were mainly associated with immune response, nutrient transport and utilization, as well as proteasome-mediated proteolysis.

Conclusion

These analysis imply that dysfunction of these biological processes or pathways of DEP in the endometrium is highly associated with early pregnancy loss. In addition, many proteins that are essential for the establishment of pregnancy showed dysregulation in the endometrium of non-pregnant ewes. These proteins, as potential candidates, may contribute to early pregnancy loss.

Electronic supplementary material

The online version of this article (doi:10.1186/s40104-015-0017-0) contains supplementary material, which is available to authorized users.