Cystic Degeneration in Porcine Ovaries - First Communication: Morphology of Cystic Ovaries, Interpretation of the Results

Contents: The morphological appearance of ovarian structures in 79 sows with ovarian cysts was examined. Sixty sows had been culled due to sterility; 19 sows had been repeatedly investigated clinically before slaughter. It was found that cystic ovaries without corpora lutea (CL) had the largest diameter and volume (151.3 cm³), in comparison to both cystic ovaries with CL (85.7 cm³) and normalovaries (16.9 cm³). The number of cysts on ovaries without corpora lutea was approximately twice as high as on the ovaries with CL (23.3 vs 12.1 cystslsow; p < 0.001). A combined form of degeneration consisting of large (> 1.5 cm in diameter) and small (1.5 em in diameter) cysts was found in 63% of the animals, whereas degeneration with only small cysts occured in 8% of the cases. Dark CL, interpreted as being functional, were mainly present in animals with up to 10 cysts per sow (24 out of 27 sows, p < 0.001); pale CL, interpreted as being weakly functional, were mainly found on ovaries with more than 10 cysts (19 out of 23 sows; p < 0.001). Based on macroscopical differentiation cyclic activity was found to be dependent to the number of cysts. Regular or irregular estrous cycles were observed in oligocystic animals (10 cysts/animal) with functional CL (58%). Up to 75% of the animals with pale or absent CL showed no estrous activity.     

Estradiol-17β, Progesterone and Testosterone Plasma Concentrations during Estrus in the Bitch

Veterinary Research Communications - Rota, A., Veronesi, M.C., Volpe, S., Riccardi, A. and Battocchio, M., 2007. Estradiol-17β, progesterone and testosterone plasma concentrations during...     

Involvement of Plasma Progesterone, Oestradiol-17β and Cortisol in Ovulatory Response to Gonadotropin-releasing Hormone in Dairy Cows with Cystic Follicles

The aim of the present study was to examine the plasma concentrations of progesterone, oestradiol-17beta and cortisol in the cows with cystic follicle and to examine its relationship with the ovulatory response to gonadotropin-releasing hormone (GnRH). Eighty-five post-partum Holstein-Friesian cows with cystic follicles regardless of the presence of corpus luteum were studied. Follicular size, presence of corpus luteum and occurrence of ovulation were checked by palpation per rectum. Blood collection and palpation per rectum were conducted on days 0, 7 and 14. Gonadotropin-releasing hormone was administered at day 7. Plasma concentrations of progesterone, oestradiol-17beta and cortisol were determined. Progesterone concentrations of <3.2, 3.2-4.8 and >4.8 nmol/l were defined as low, intermediate and high, respectively. Sixty-three (74.1%) of 85 cows showed low (<3.2 nmol/l =1 ng/ml) progesterone concentrations on day 0. Only 40 (47.1%) of them showed low-low pattern of progesterone at days 0 and 7. In 27 (31.8%) of them, progesterone concentration had increased by day 14. Of 22 cows having high progesterone concentration (>/=4.8 nmol/l) on day 0, corpus luteum was not detected in 18 cows (21.2%). Only in 10 cows, cystic follicle disappeared after GnRH administration. However, only one of 27 cows in which progesterone pattern was low-low-high at days 0, 7 and 14 experienced ovulation of the cystic follicle. Significantly lower oestradiol-17beta concentration was found on day 7 in cows showing a low-low-low pattern than a low-low-high pattern of progesterone (43.0 +/- 4.6 vs 55.8 +/- 2.8 pmol/l, p < 0.05). There was no significant difference in cortisol concentration on any days (days 0, 7 and 14) between cows showing a low-low-low and low-low-high pattern of progesterone. These results suggest that approximately one-fifth of cows diagnosed to have ovarian cysts possess luteal cysts and that a high oestradiol-17beta concentration at the time of GnRH administration is involved in the subsequent ovulation of the follicle, although ovulated follicle may not be cystic.     

Changes in Plasma Concentrations of LH, FSH, Estradiol 17-β and Progesterone During Oestrus in Mithun (Bos frontalis)

The objective of the present study was to establish the changes in plasma concentrations of LH, FSH, estradiol 17-beta (E2) and progesterone (P4), as well as to understand their temporal relationships during oestrus in mithun (Bos frontalis). The experiment was conducted on 11 mithuns during third or fourth postpartum oestrous cycle. Since oestrus onset the jugular vein blood samples were collected every 2 h for 72 and 96 h, respectively from the animals without and with standing heat. The LH, FSH, E2 and P4 concentrations were estimated in plasma. The P4 concentration was fluctuated throughout the oestrus period and the average P4 concentration was found significantly (p<0.05) lower on the day of oestrus onset. The multiple rises in LH and FSH concentrations above the basal level in spike like fashion were observed throughout the oestrus period irrespective of the occurrence of standing heat. A significant (p<0.01) gradual increase in the average daily E2 concentration was observed till day 2 following oestrus onset irrespective of the occurrence of standing heat. A significant (p<0.05) simultaneous increase in LH, FSH and E2 concentrations and a transient increase in P4 concentration at approximately the time of standing heat onset were observed. During investigation a definite temporal coupling between LH and FSH rises was absent throughout the oestrus period. The results suggest that (1) the multiple short-duration low-amplitude LH and FSH surges during oestrus may be crucial for the final maturation of ovulatory follicle and subsequent ovulation in mithun; (2) a differential mechanism for controlling LH and FSH secretions probably exists in mithun.     

Age-related Variation of Plasma Concentrations of Skatole, Androstenone, Testosterone, Oestradiol-17β, Oestrone Sulphate, Dehydroepiandrosterone Sulphate, Triiodothyronine and IGF-1 in Six Entire Male Pigs

This study describes the age-related variation in boar taint compounds, skatole and androstenone, and testosterone, oestradiol-17 beta (E17 beta), oestrone sulphate (ES), dehydroepiandrosterone sulphate (DHEAS), triiodothyronine (T(3)) and insulin-like growth factor-1 (IGF-1) in six boars. Three pairs of littermates of crossbred entire male pigs (from three Yorkshire x Duroc dams and one Hampshire sire) were included. Blood samples were taken at the age of 9-15 weeks and thereafter at weekly intervals from the age of 20-32 weeks. Plasma concentrations of skatole, androstenone, testosterone, E17 beta, ES, DHEAS, T(3) and IGF-1 were measured. We found that skatole levels in boars increased at the age around puberty after an increase in the levels of testicular steroids. Levels of skatole were not associated with the levels of sex steroids, T(3) and IGF-1. However, the increased level of testicular steroids is probably the underlying factor needed for high skatole levels to occur although the specific mechanism leading to increased skatole levels remains unknown.     

17β‐Estradiol,Progesterone and Testosterone Concentrations in Cystic Fluids and Response to GnRH Treatment after Emptying of Ovarian Cysts in Dairy Cows

The aim of this study was to determine possible links between steroidogenic activity of single ovarian cysts and response to intramuscular treatment with 20 μg of buserelin (GnRH‐analogue) after cyst emptying, in pluriparous Friesian cows bearing a singleton cyst treated not earlier than 55 days post‐partum. Progesterone, 17β‐estradiol and testosterone were determined in cystic fluids collected by needle aspiration of the cyst. Of the cows, 75.6% began ovarian cyclicity within 30 days after treatment with a conception rate of 64.7%. In this study it was found that as progesterone concentration in cystic fluids rose, the number of positive responses to the treatment fell.     

The Content of β-endorphin-like Immunoreactivity in Porcine Corpus Luteum and the Potential Roles of Progesterone, Oxytocin and Prolactin in the Regulation of β-endorphin Release from Luteal Cells in vitro

The amount of β‐endorphin‐like immunoreactivity (β‐END‐LI) in porcine corpora lutea from several stages of the oestrous cycle and the effects of progesterone, oxytocin, and prolactin on β‐END‐LI secretion in vitro by luteal cells were studied. Porcine corpora lutea obtained on days 1–5, 6–10, 11–13, 14–18, and 19–21 of the cycle were used to prepare extracts for β‐END‐LI determination. Additionally, corpora lutea from days 11–13 and 14–18 were enzymatically dissociated and isolated luteal cells were used for further study of β‐endorphin secretion in vitro. Cells were cultured in serum‐free defined M 199 medium (10⁶ cells/ml) at 37°C under 5% CO₂ in air, for 12 h. The influences of the following factors on β‐END‐LI secretion by luteal cells were tested: progesterone (10^–9, 10^–7 and 10^–5M ), oxytocin (0.01, 0.1, 1 and 10 ng/ml), and prolactin (0.1, 1, 10 and 100 ng/ml). The β‐END‐LI contents in extracts and media were measured by radioimmunoassay. The tissue concentration of β‐END‐LI was lowest on days 1–5 of the cycle (0.35 ± 0.03 ng/g wet tissue). Subsequently, it constantly increased to the highest value on days 14–18 (16.58 ± 0.52 ng/g wet tissue) and on days 19–21 it declined (11.10 ± 0.52 ng/g wet tissue). Progesterone at a low dose (10^–9 M ) resulted in significant (p < 0.05) increases and decreases in β‐END‐LI secretion by luteal cells from days 11–13 and 14–18, respectively. Higher doses of progesterone (10^–7 and 10^–5 M ) had no effect on β‐END‐LI release, compared with the control group. All dose‐levels of oxytocin used decreased β‐END‐LI secretion by luteal cells on days 11–13 and 14–18 of the cycle. Prolactin at doses of 0.1 and 1 ng/ml on days 11–13, and all doses tested on days 14–18 resulted in decreases in β‐END‐LI release from luteal cells. These results document evident changes in β‐END‐LI content in the pig corpus luteum during its development and indicate the potential roles of progesterone, oxytocin, and prolactin in luteal cell secretion of β‐END‐LI.     

Effect of Oestrous Synchronization with Estradiol 17β and Progesterone on Follicular Wave Dynamics in Dairy Heifers

An experiment was designed to evaluate the effects of estradiol‐17β (E17β) on follicular wave dynamics and ovulatory response in Holstein heifers receiving either a progestogen ear‐implant (Crestar^®; Intervet International b.v. Boxmeer, The Netherlands) or an intravaginal progesterone‐releasing device [controlled internal drug release‐bovine device (Eazibreed, CIDR‐B^®; Bodinco BV, Alkmaar, The Netherlands)]. For comparison, another group of heifers was also synchronized using Crestar plus an injection of estradiol valerate (EV) and norgestomet as recommended by the pharmaceutical company. Twenty 20–22‐month‐old cycling Holstein heifers were allocated to one of the following treatment groups at random stages of the oestrous cycle: (I) simultaneous insertion of Crestar and intramuscular injection of 3 mg norgestomet and 5 mg EV (Crestar 9 + EV 9); (II) simultaneous insertion of Crestar and intramuscular injection of 5 mg E17β (Crestar 9 + E17β 9); (III) insertion of Crestar followed 2 days later by intramuscular injection of 5 mg E17β (Crestar 9 + E17β 7); or (IV) insertion of CIDR‐B device followed 2 days later by intramuscular injection of 5 mg E17β (CIDR 9 + E17β 7). The CIDR‐B or Crestar implants were removed after 9 days and all heifers received 500 μg Cloprostenol (Estrumate^®, Pitman‐Moore Nederland BV, Houten, The Netherlands). Ovarian ultrasonographic examinations were performed once daily during the synchronization period using a B‐mode scanner equipped with a 7.5 MHz linear‐array transrectal transducer. In addition, heifers were scanned every 12 h after implant/device withdrawal until 3 days after ovulation in order to monitor follicular activity, detect ovulation and subsequent early luteal formation. Detection of oestrus was performed every 6 h for 4 days after device/implant removal. Oestrus was observed 24–32 h before ovulation in all heifers. The mean hours interval from treatment withdrawal to ovulation was not significantly different (84.0 ± 16.5, 77.6 ± 4.1, 73.6 ± 4.1 and 64.0 ± 4.4 h for treatments I, II, III and IV, respectively; p > 0.1). However, the variance for heifers treated with EV + norgestomet was significantly larger (Levene’s Test; p < 0.01) than those treated with E17β. All E17β treatments resulted in dominant follicle suppression and a new wave emerged 4.1 days after treatment compared with 6.6 days for the EV + norgestomet treatment (p < 0.05). The time from emergence of the new ovulatory wave to ovulation was longer for the new wave that emerged after E17β treatment (9.2 ± 0.3 days) than after EV + norgestomet treatment (6.9 ± 0.4 days; p < 0.05). The results of this study suggest that the four treatments used were effective in inducing synchronous behavioural oestrus and ovulation. However, a higher degree of oestrus and ovulation synchrony was observed in heifers treated with E17β than in heifers treated with EV + norgestomet. Synchronization treatments with exogenous E17β or EV + norgestomet at the time of progestin device insertion (Crestar or CIDR‐B) or 2 days later in heifers can regulate a different emergence pattern of ovarian follicular development in randomly cyclic heifers. The E17β was effective in inducing follicular suppression and resulted in the consistent emergence of a new follicular wave.     

Effects of Mating on Plasma Concentrations of Testosterone, Cortisol, Oestrone Sulphate and 15-Ketodihydro-PGF2α in Stallions

Very little information is available regarding the physiological mechanisms involved in the normal sexual activity in the stallion and, in particular, the endocrine control of reproduction is still not clearly understood. This experiment was designed to determine the short-term effect of sexual stimulation on plasma concentrations of testosterone, cortisol, oestrone sulphate and 15-ketodihydro-PGF(2alpha) in stallions. Semen samples were collected from 10 lighthorse stallions of proven fertility using a Missouri model artificial vagina. At the same time, blood samples were collected from the jugular vein with heparinized tubes, 20 and 10 min before oestrous mare exposure, at exposure and 10, 20, 30 min after dismounting. Testosterone concentrations showed a sharp rise 10 min after mating (p < 0.001), reached a plateau, and then showed a further increase 30 min after mating (p < 0.001). Cortisol concentrations increased 10 min after mating (p < 0.001) and remained at high levels in the subsequent samples taken. A peak of oestrone sulphate was observed 10 min after mating (p < 0.001). 15-Ketodihydro-PGF(2alpha) concentrations decreased rapidly at the moment of the exposure of the stallions to an oestrous mare (p < 0.05), returned to pre-mating concentrations and then decreased again 30 min after mating (p < 0.05).     

Immunohistochemical Localization of Oestrogen Receptors α and β, Progesterone Receptor and Aromatase in the Equine Placenta

The functions of placental oestrogens during equine pregnancy are still unclear. Yet, they may act predominantly as local regulators of growth and differentiation in the microplacentomes. Thus, expression patterns of oestrogen receptors (ERs) α and β were investigated in the microcotyledonary placenta from pregnant mares at 110, 121, 179, 199 and 309 days of gestation by immunohistochemistry. In microplacentomes, both the ER isoforms were detected in trophoblast (T) cells, chorionic villous stroma (FS), microcaruncular epithelium (ME) and microcaruncular stroma (MS). Proportions of positive cells were 38–91% (T), 11–41% (FS), 55–89% (ME), 17–51% (MS) for ERα and 66–76% (T), 21–37% (FS), 41–68% (ME) and 24–55% (MS) for ERβ. Between days 110 and 199, proportions of cells positive for progesterone receptor (PR) varied between 19% and 62% (T), 3% and 50% (CS), 15% and 46% (ME), and 4% and 33% (MS). At day 309, PR was virtually absent in T, CS and ME (percentages < 0.1), whereas in MS 14.3% of cells were still positive. The expression of ERs and PR in equine microplacentomes gives evidence for a role of placental steroids as regulators of placental growth, differentiation and function. The detection of ERα, ERβ and PR in foetal and maternal vascular tissue suggests that placental steroids are also involved in the control of placental angiogenesis and /or vascular functions. The co-localization of ERs with aromatase in T suggests auto- or intracrine functions of oestrogens in this cell type.     

Vitamin A and β-Carotene Levels in Plasma, Corpus Luteum and Follicular Fluid of Cyclic and Pregnant Cattle

This study was carried out to examine the relationship between the corpus luteum (CL) weight, CL and follicle diameters and progesterone, β‐carotene and vitamin A levels in reproductive organs of cattle obtained from the slaughterhouse. The β‐carotene and vitamin A levels were determined in plasma, CL and follicular fluid (FF) using a spectrophotometric method at different stages of the oestrous cycle (n=40) and at 3–6 months of pregnancy (n=10). The diameters of the CL and follicle were measured using ultrasonography. Plasma progesterone concentrations were determined by an enzyme immunoassay method. The vitamin A levels of the plasma, CL and FF were not related to each other. The highest plasma vitamin A levels were observed in the proestrus and oestrus, at which periods follicular activity dominates. The vitamin A levels in the CL and FF were negatively related to the weight and diameter of the CL and the diameter of follicle, respectively. In contrast to vitamin A, β‐carotene concentrations of plasma, CL and FF were significantly correlated with each other. The highest β‐carotene levels in the plasma, CL and FF were found during pregnancy when there is maximal luteal function, and the β‐carotene level of the CL was significantly correlated with the weight and diameter of CL. Furthermore, the intrafollicular β‐carotene level was negatively correlated with the follicle diameter. There was a positive correlation between plasma progesterone level and the weight and diameter of the CL, but a negative correlation between plasma progesterone level and follicle diameter. Moreover, plasma, FF and CL β‐carotene levels were positively correlated with plasma progesterone levels. This study revealed that β‐carotene levels in the plasma, CL and FF were influenced by the stage of the oestrous cycle or the pregnancy and were related to bovine luteal function without depending on vitamin A.     

24-Hour Secretion Patterns of Plasma Oestradiol 17β in Pony Mares in Late Gestation

The mare exhibits nocturnal uterine contractions in the last 6 days of gestation. It is hypothesized that estradiol 17β (O17β) may be associated with the nightly increase in uterine contractions. The 24‐h secretion pattern of plasma O17β was measured in 3 pony mares in late gestation to identify changes in release as the mare neared parturition. Blood was collected weekly at 08:00 hours beginning on day 240 and every third day from day 330 until delivery. Serial blood samples were collected from each mare every 30‐min for 24‐h beginning on gestation day 310 and every sixth day thereafter until parturition. Concentrations of O17β were elevated at night with lowest concentrations occurring directly before sunset (p < 0.01). The natural log of the variance was increased at sunset (p < 0.01) and was decreased during the 6‐h period immediately after sunrise. This pattern was especially evident in the 6 days that preceded parturition. The contrast between nocturnal and daytime concentrations of O17β in the last 6 days of gestation may contribute to night‐time delivery in the mare.     

Concentrations of Oestrone and Oestradiol-l 7β in the Peripheral Plasma of the Nanny Goat during the Oestrous Cycle, Gestation and Puerperium

The changes in the plasma levels of oestrone sulphate and oestradiol-17β during the oestrous cycle, gestation and puerperium in the goat are described. The oestrone sulphate concentrations remained fairly constant (250–350 pg/ml) throughout the oestrous cycle until day 20 when a sharp increase of the oestrone sulphate plasma levels occurred in pregnant goats which became significantly different at day 38 of gestation from nonpregnant values. Oestradiol-17β plasma levels were significantly lower at days 17–20 in pregnant than in nonpregnant does. Oestrone sulphate and oestradiol-17β concentrations rose until the 12th week of gestation and then declined to about 50% of the former ranges of concentrations before rising again to high values at weeks 17–20 of gestation. Increasing plasma levels of oestrone sulphate and oestradiol-17β were determined during the last ten days preceding parturition. The concentrations of oestrone sulphate returned to basal levels by the 2nd—4th day post partum whereas oestradiol-17β values reached base values 24 hours after parturition. Both oestrogen concentrations remained constant during the puerperium until day 51 post partum .     

The Relationship Among Vitamin C, β‐carotene,Vitamin A,Progesterone and Oestradiol 17‐β Concentrations in Plasma and Cyst Fluid of Holstein Cows with Ovarian Cyst

The aims of this study were to determine the concentrations of the progesterone, oestradiol‐17‐β, vitamin A, C and β‐carotene in plasma and cyst fluid and to relate these values with cystic diameter and membrane thickness of Holstein cattle with ovarian luteal cyst. 1650 Holstein cows were examined for the presence of the ovarian cyst and luteal and follicular cystic ovaries were obtained following slaughtering in personal slaughterhouse in Konya‐Turkey. 15 Luteal and 15 follicular cystic ovaries were distinguished by rectal palpation and by post mortem ultrasonographic examination. Plasma and cyst fluid, hormone and vitamin analyses were carried out by EIA method and spectrophotometric measurement respectively. Although there was no relationship between β‐carotene and vitamin A in plasma and cyst fluid of both cyst type and hormone concentrations, the vitamin C concentration of cyst fluid was found significantly higher in luteal cyst than in follicular cyst. Moreover, there is a positive correlation among values of the vitamin C concentrations of cyst fluid and cystic membrane thickness, plasma and the cyst fluid progesterone concentrations, but there is a negative correlation among the vitamin C concentrations of cystic fluid and oestradiol 17β levels of plasma and cyst fluid. In conclusion, vitamin C concentration of cyst fluid supported ultrasonographic and endocrinologic findings. Also, it can be postulated that vitamin C is probably effective on progesterone synthesis in the luteal tissue of cyst.     

The Effect of Piglet Expulsion in the Sow on Plasma Cortisol, Adrenocorticotropic Hormone and β-endorphin

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Previous studies have shown an increase in plasma cortisol in gilts over farrowing irrespective of environment suggesting that factor(s) associated with parturition itself cause physiological stress. Factors involved in mediating the hypathalomo–pituitary–adrenal (HPA) axis at parturition are not well understood. This study examines the effect of piglet expulsion on the pituitary–adrenal axis by measurement of plasma cortisol, adrenocorticotropic hormone (ACTH) and β-endorphin. The effect of farrowing environment in modulating the acute response to piglet expulsion is also investigated. Twelve second parity sows, with indwelling jugular catheters, were moved into either a farrowing crate or a straw-bedded pen 5 days before their expected parturition date (EPD). Blood samples were taken from each sow during a prefarrowing baseline period and then rapid samples (2.5 mins) were taken for 10 min following the birth of two piglets. No effect of environment was found on any of the hormonal variables which reinforces the hypothesis that the physiological stress seen in parturient pigs is due to some intrinsic factor of parturition. Plasma cortisol, ACTH and β-endorphin did not change significantly in the period following piglet expulsion suggesting that individual piglet expulsions do not play a major role in 'parturition stress'. There was however, an increase in plasma cortisol, ACTH and β-endorphin in response to increasing piglet number which is consistent with previous studies of general farrowing in which cortisol increased as farrowing progressed. Therefore this study reinforces the hypothesis that physiological stress increases with ongoing parturition although this does not appear to be a result of piglet expulsion. The potential role of other factors which may be involved in causing 'parturition stress' should be investigated.