Blood plasma concentrations of oestradiol-17β, testosterone and testosterone/oestradiol ratio in dogs with neoplastic and degenerative testicular diseases

Oestradiol-17β and testosterone blood plasma concentrations were measured in dogs with Leydig-cell tumours (n=20), Sertoli-cell tumours (n=6), seminomas (n=9), unilateral inguinal cryptorchidism (n=7), abdominal cryptorchidism (n=9, one bilateral), degenerate scrotal testicles (n=6, two bilateral), and animals with normal scrotal testicles (n=20). The testosterone/oestradiol ratio (testosterone concentration [ng/mL] × 100/oestradiol concentration [pg/mL]) was calculated.A considerably higher oestradiol concentration was found in dogs with Sertoli-cell tumours (29.0, 14.4–48.3 pg/mL; median, minimum–maximum; P=0.0256, Mann–Whitney test) and lower oestradiol levels were found in animals with seminomas (12.0, 3.4–17.6 pg/mL; P=0.0025) compared to the healthy control group (18.0, 8.6–31.5 pg/mL). Testosterone concentration was decreased in dogs with Sertoli-cell tumours (0.08, 0.03–0.77 ng/mL) when compared to the control group (1.95, 0.05–3.70 ng/mL; P=0.0012). Testosterone/oestradiol ratios differed from the control (9.6, 0.58–35.8) only in dogs with Sertoli-cell tumours (0.32, 0.06–2.80; P=0.0005). Clinical signs of feminization were observed in five dogs with Sertoli-cell tumour and one dog with a Leydig-cell tumour, and were more often associated with decreased testosterone/oestradiol ratios than with an increased oestradiol-17β concentration.     

Relationship between testicular transferrin and plasma estradiol-17beta concentrations of dogs with azoospermia and dogs with sertoli cell tumors.

Testicular Transferrin (Tf) and peripheral plasma estradiol-17beta (E2) concentrations were measured in 3 dogs with azoospermia (AZ dogs), 3 dogs with Sertoli cell tumors (SC dogs), and 5 normal male Beagles. The mean Tf concentrations in the testes of the AZ dogs and the affected testes of the SC dogs, and the plasma E2 concentrations in both these groups of dogs were significantly higher than the values in normal dogs (P<0.05, 0.01 and 0.01, respectively). Therefore, excessive E2 secretion by hyperfunctioning Sertoli cells is thought to have caused the azoospermia in the 3 dogs.     

Effect of testosterone and oestradiol-17beta on canine hair follicle culture

Skin biopsies were taken from four body sites (head, thorax, flank and perineum) of three male entire Beagles and the primary hair follicles were isolated. Culture conditions were established to keep the hair follicles growing for up to 7 days. Additionally, hair follicles were incubated in supplemented medium (containing insulin, transferrin, glutamine and sodium selenite) with or without the addition of testosterone (T) (1, 10 or 100 ng/ml) or oestradiol-17beta (E2beta) (0.01, 0.1 or 1 ng/ml), respectively and the daily growth of hair follicles was measured. In vitro daily growth of hair follicles from the thorax was stimulated by the low concentration of both hormones, but the growth of those from the flank was inhibited by the high concentration of both hormones. Hair follicles from the head were positively influenced by the lowest concentration of T and the medium concentration of E2beta. The daily growth of hair follicles from the perineum was not significantly influenced by either hormone.     

Serum testosterone and estradiol 17-beta concentrations in 15 dogs with perineal hernia

Serum testosterone and estradiol 17-beta concentrations, and serum testosterone-to-estradiol ratio were evaluated in 15 dogs (greater than or equal to 5 years old) with perineal hernia (9 sexually intact males and 6 castrated males) and in 9 clinically normal sexually intact male dogs greater than or equal to 5 years old. There was no significant difference in serum testosterone-to-estradiol ratio between sexually intact male dogs with perineal hernia and clinically normal sexually intact male dogs. In castrated dogs with perineal hernia, serum testosterone concentration and testosterone-to-estradiol ratio were significantly (P less than 0.05) lower, compared with those values in sexually intact dogs with perineal hernia and in clinically normal sexually intact male dogs. There was no significant difference in serum estradiol 17-beta concentration among sexually intact male dogs with perineal hernia, castrated dogs with perineal hernia, and clinically normal sexually intact male dogs. Serum testosterone and estradiol 17-beta concentrations in dogs with perineal hernia did not differ from those values in clinically normal male dogs of the same age. Castration cannot be recommended for the treatment of perineal hernia unless a castration-responsive contributing factor such as prostatomegaly is identified, unless the pelvic diaphragm of dogs with perineal hernia has high sensitivity to normal or low serum testosterone and estradiol 17-beta concentrations, or unless there is documentation that other androgens and/or estrogens are involved.     

Response of anoestrous ewes pre-treated with a single dose of oestradiol-17 beta, or progesterone and oestradiol-17 beta, to the introduction of rams and ewes in oestrus

AIMS: To determine if the frequency distribution of time to oestrus observed after the introduction of rams, and additional ewes in oestrus, is affected by the administration of oestradiol- 17beta. To determine if the fertility following mating at the first induced oestrus in progesterone-primed ewes may be increased with administration of oestradiol-17beta. METHODS: In Experiment 1, ewes received 40 mug oestradiol- 17beta 3 (E3; n=61) or 5 (E5; n=56) days before the introduction of rams; 61 ewes were controls (C1). In Experiment 2, a controlled internal drug-releasing (CIDR) device was inserted in 95 ewes on Day -5 (Day 0 = introduction of rams). In addition, 47 ewes received oestradiol-17beta on Day 0 (CE), and the remaining 48 were controls (C2). In both experiments, additional ewes, brought into oestrus between Days 0 and 2 by hormonal treatments, were introduced with the rams. Onset of oestrus was estimated by visual observation of ewes marked by rams. Pregnancy status of ewes in Experiment 2 was determined using ultrasound, 30 days after oestrus. RESULTS: The total number of ewes detected in oestrus in Experiment 1 was similar between the three groups. The frequency distribution of ewes in oestrus in the E5 group differed from that of ewes in both C1 (p=0.05) and E3 (p<0.001). A similar number of ewes were detected in oestrus between Days 16-20, but the proportion of ewes in oestrus between Days 21-26 was greater for ewes in C1 than E5 or E3. Mean interval to the onset of oestrus was shorter in C1 than E3 which was shorter than E5 ewes (p<0.001). In Experiment 2, the total number of ewes in oestrus were similar between groups. Ewes in oestrus between Days 1-4 (65% and 64% for C2 and CE, respectively) and conception rates over the same period (81% and 80% for C2 and CE, respectively) were also similar between groups. CONCLUSIONS: Administration of oestradiol-17beta to anoestrous ewes altered the pattern of onset of oestrus, but it did not affect submission or conception rates, regardless of whether or not a short period of progesterone priming with a new CIDR device was included prior to the introduction of rams.     

Effects of oestradiol-17beta and testosterone on progesterone production in the cultured granulosa cells of Japanese quail

In order to study the effects of steroid hormones on steroidogenesis in the avian ovary, quail granulosa cells were cultured with follicle stimulating hormone (FSH), oestradiol-17beta or testosterone. The progesterone content of the medium during the culture period of 66 h and the following 3 h of incubation with luteinising hormone (LH), was measured by radioimmunoassay. When FSH, oestradiol-17beta or testosterone were added during the 66 h culture, subsequent progesterone production by the cells during 3 h of incubation with LH was significantly increased. However, testosterone also stimulated progesterone production in the medium during the 66 h culture period, whereas FSH oroestradiol-17beta did not. Addition of staurosporine during culture inhibited both LH-stimulated progesterone production and testosterone-stimulated progesterone production. These results indicate that the processes during which granulosa cells acquired responsiveness to LH, and testosterone stimulates progesterone production might both be mediated by a staurosporine-sensitive protein kinase C-dependent pathway in quail granulosa cells.     

Pfaffia paniculata-induced changes in plasma estradiol-17beta, progesterone and testosterone levels in mice

The present study undertook chemical analysis of components of Pfaffia paniculata roots. In addition, an animal experiment was conducted in which mice had ad libitum access to water enriched with powdered P. paniculata root for 30 days. Changes in plasma concentrations of estradiol-17beta and progesterone in female mice and of testosterone in male mice were ascertained. The results revealed that P. paniculata roots contain two types of phytosteroids, beta-sitosterol and stigmasterol, in addition to other compounds such as pfaffic acid, allantoin, saponins, beta-sitosteryl-beta-D-glucoside, and stigmasteryl-beta-D-glucoside. Regarding changes in plasma concentrations of hormones, levels of the sex hormones estradiol-17beta, progesterone and testosterone were clearly higher for mice that drank P. paniculata root-enriched water than for mice that drank plain water. Powdered P. paniculata root is easily dissolved in feed or water, and as no adverse reactions were seen in mice within 30 days of oral intake, consumption of P. paniculata for long periods of time appears safe.     

Post-partum, ovarian function in dairy cows as revealed by concentrations of oestradiol-17 beta and progesterone in defatted milk

Concentrations of oestradiol-17 beta and progesterone in defatted milk have been used to study the post-partum restoration of ovarian function in 52 autumn-calved dairy cattle. In 32 cows the first preovulatory peaks in oestradiol-17 beta concentration (indicative of imminent ovulation) were less than or equal to 15 days and in 49 cows less than or equal to 49 days post partum. Delay to first ovulation was mainly due, not to failure of ovarian secretion of oestradiol-17 beta at preovulatory level, but to failure of oestradiol-17 beta at this level to exert its normal preovulatory function. Longer intervals to first ovulation were associated with longer intervals from calving to the clearance of placental oestradiol-17 beta, high peak milk yields and high body weight losses, suggesting that in high-yielding dairy cows these factors (themselves inter-related) may be associated with others, which are the immediate cause of the inhibition of the normal, preovulatory function of oestradiol-17 beta. Forty-eight per cent of cows had short, first ovulation cycles. Ovarian function, comprising oestradiol-17 beta secretion at preovulatory level, normal preovulatory function of oestradiol-17 beta and normal corpus luteum progesterone secretion were almost fully restored in this herd by the 49th day post partum.     

Exfoliate vaginal cytology and plasma concentrations of progesterone, luteinising hormone and oestradiol-17sZ during oestrus in the bitch

Oestrus was characterised in nine German shepherd bitches by measuring plasma levels of luteinising hormone (LH), progesterone (P) and oestradiol-17ß (O). Hormone concentration data were compared with the eosinophilic index (EI) obtained from exfoliate vaginal cytology. Maximal values of O and LH occurred before the maximal EI value. The EI peak followed the O peak, with a two day delay. LH increased during the first 24 hours after the maximum 0 value, and P levels were then slightly higher than basal.     

Changes in plasma testosterone and testicular transferrin concentration, testicular histology and semen quality after treatment of testosterone-depot plus PMSG to 3 dogs with asthenozoospermia

Three dogs diagnosed as having asthenozoospermia were given three intramuscular injections of 50 mg testosterone(T)-depot plus 250 IU pregnant mare serum gonadotropin (PMSG) at 2-week intervals, and their plasma T and testicular transferrin (Tf) concentrations, testicular histology, and semen quality were examined during the period of hormone therapy. Plasma T concentrations temporarily increased, and there was a slight improvement in spermatogenesis. Increased Tf concentrations suggested that Sertoli cell function in all three dogs was promoted by hormone treatment. The results showed that semen quality, especially the percentages of motile sperm and abnormal sperm, were improved between 1 and 5 weeks after the start of T-depot plus PMSG treatment.     

High plasma estradiol-17beta levels in dogs with benign prostatic hyperplasia and azoospermia

The semen quality of 22 dogs (4 to 7 years old) with benign prostatic hyperplasia (BPH) was examined at the hospital of our university, and 4 of the 22 BPH dogs were diagnosed as azoospermic. The mean peripheral plasma estradiol-17beta (E2) level (17.3 pg/ml) of the 18 BPH dogs with spermatogenic function was higher than that of 5 normal male dogs and their mean T level (1.7 ng/ml) was lower. The mean E2 level (27.3 pg/ml) of the 4 BPH dogs with azoospermia was significantly higher than the value in the BPH dogs with spermatogenic function (P<0.01), and the mean T level (1.1 ng/ml) was significantly lower (P<0.05). Five normal male dogs were given 10 intramuscular injections of estradiol benzoate (E2B) 5 microg/kg, at 3-day intervals to investigate the relationship between high plasma E2 levels and the cause of the BPH and azoospermia. Their testes and prostates were measured and biopsied both before and 30 days after the start of E2B injections. At 30 days after the start of the E2B injections, the mean peripheral plasma T levels had decreased by half, and the mean testicular volume had decreased to 88% of original volume. The numbers of spermatocytes, spermatids, and spermatozoa in the seminiferous tubules of all of the dogs were significantly lower (P<0.05, 0.01). In addition, the mean prostatic volume increased to 130%, the mean height of the glandular epithelium decreased, and the glandular lumen became increased in diameter. These findings indicate that both BPH and serious spermatogenic dysfunction may be simultaneously induced by protracted high plasma E2 levels in dogs.     

Serum concentrations of immunoglobulins G, A, and M in dogs with neoplastic disease

The concentrations of the three major classes of immunoglobulins (Ig) were determined in canine serum by single radial immunodiffusion against calibrated standards. Serum samples were collected from 121 dogs categorized into 3 groups: normal dogs (n = 34), those with lymphosarcoma (n =41), and those with malignant, solid neoplasms other than lymphosarcoma (n = 46). The mean value for serum IgM concentration in the group of dogs with lymphosarcoma was significantly (P less than or equal to 0.02) higher than was the mean IgM concentration of the normal dogs. Dogs with neoplasms other than lymphosarcoma had significantly increased serum concentrations of IgG and IgM antibodies. There was no significant difference in serum IgA concentration among the three groups. A wide range of Ig concentrations was in the serum of clinically normal dogs, as well as in dogs with neoplastic diseases. Although individual dogs with neoplasms had low serum Ig concentrations, the 81 dogs with neoplastic disease were generally able to synthesize Ig.     

Changes in concentrations of plasma immunoreactive follicle-stimulating hormone, luteinizing hormone, estradiol-17beta, testosterone, progesterone, and inhibin in heifers from birth to puberty

This study was designed to clarify the characteristics of changes in plasma concentrations of reproductive hormones in heifers from birth to puberty. Weekly or daily hormonal changes were observed in 39 heifers. Daily changes in the concentration of follicle-stimulating hormone (FSH) demonstrated a consistent cycle of hormone changes over a 7- to 8-day period in heifers from approximately 10 days to 9 months old. Weekly changes in reproductive hormones showed that there were three brief periods in heifers between birth and puberty in which dramatic changes occur. The first period was the first week after birth, during which a reciprocal relationship between steroid hormones and gonadotropins was observed. At birth, the concentrations of steroid hormones were higher than those at any other age. These hormone levels rapidly decreased within the first week after birth. Gonadotropin levels, however, increased from birth to 1 week of age. The second period of major change was at approximately 4 weeks of age when there was an increase in the concentrations of luteinizing hormone (LH), estradiol-17beta, testosterone, and immunoreactive inhibin. The third period was the last 5 weeks before the first ovulation, when there was an increase in the concentrations of estradiol-17beta followed by an increase in (LH). These results suggest that regular hormone changes start from 10 days after birth and that the periods from birth to 4 weeks of age and the last 5 weeks before the first ovulation in heifers are important to the development of reproductive functions before puberty.     

In vitro effects of prepartum concentrations of oestradiol-17 beta on cell-mediated immunity and phagocytosis by porcine leukocytes

The in vitro effect of oestradiol-17 beta (Oe2) on cell-mediated immunity (CMI) and phagocytosis by leukocytes obtained from ten and eight sows, respectively, was studied. The concentrations of Oe2 used were 300 pmol/l and 3000 pmol/l, to reflect a low and a high prepartum concentration of the hormone. The CMI was measured by lymphocyte proliferation (LP) tests performed on whole blood and purified mononuclear cells (MNC) and by estimation of the production of interleukin-2 (IL-2) in the latter type of culture. In the whole blood LP test, the mitotic response to concanavalin A (Con A) was increased by 29% (P less than or equal to 0.05) when the Oe2 concentration was 300 pmol/l. The higher concentration of Oe2 (3000 pmol/l) increased the amount of IL-2 in supernatants from Con A-stimulated MNC by 17% (P less than or equal to 0.05). The phagocytic function of polymorphonuclear leukocytes was determined in a chemiluminescence assay. In this assay, the maximal rate of phagocytosis was reduced by 12% (P less than or equal to 0.01) and the area under the curve by 10% (P less than or equal to 0.05) at an Oe2 concentration of 3000 pmol/l. The results show that prepartum concentrations of Oe2 can promote CMI in vitro, but weaken phagocytosis by porcine leukocytes.     

Concentrations of oestradiol-17 beta in plasma and milk and progesterone in plasma during the oestrus cycle and in early pregnancy in goats

Pre-ovulatory peaks in oestradiol-17 beta concentrations were observed on days 1 or 2 and post-ovulatory peaks between days 4 and 7, both in jugular venous plasma and defatted milk, day 1 being the day of the onset of oestrus in the goats. Mean values of the magnitudes of these concentration peaks and of their timing (relative to oestrus) during the oestrus cycle did not differ significantly (P greater than 0.05) from those when the goats were mated and became pregnant. Pre-ovulatory oestradiol-17 beta peaks were invariably greater than the corresponding post-ovulatory peaks, as were peak concentrations in plasma relative to those in defatted milk collected on the same day. Mean intervals between the pre- and post-ovulatory peaks in oestradiol-17 beta concentrations were respectively 4.2 days for plasma and 4.0 days for defatted milk. Concentrations of oestradiol-17 beta in jugular venous plasma and defatted milk were strongly correlated: rank correlation coefficients for the three goats studied were 0.871, 0.668 and 0.739. It is suggested that in goats, as in cattle, ovarian follicular oestradiol-17 beta secretion approaching pre-ovulatory level is restored by 4 days after oestrus and its rapid decline after this time may be due to the inhibitory influence of the rapidly rising plasma progesterone concentration.