Aetiological analysis and diagnosis of reproductive disorders in male dromedary camels.

The purpose of this paper was to analyse the aetiology and methods of diagnosing reproductive disorders in male dromedary camels. Male camel infertility manifests as one of three conditions: post-coital infertility (IG), inability to copulate (IC) and lack of sexual desire (LSD). IG is mainly a testicular disorder that is linked to a deteriorated seminogram, arrested spermatogenesis, Sertoli cell-only syndrome and testicular degeneration. For IG diagnosis, semen analysis, testicular biopsy and fine-needle aspiration are gold standards. Testicular ultrasonography was generally inefficient. High serum FSH was found in IG camels with oligo- and azoospermia, implying primary spermatogenesis defects. The testis-expressed protein (TEX101) and the epididymis-expressed protein (ECM1) are reliable biomarkers for distinguishing between obstructive and non-obstructive azoospermia. IC manifests in two forms: phimosis (PHI) and erectile dysfunction (ED). PHI is frequently linked to preputial and penile pathologies, as well as leucocytosis, neutrophilia and elevated nitric oxide metabolites. The majority of camels with ED have normal genital organs, and the condition is associated with an increase in cardiac troponin I. LSD is a rare disorder brought on by hormonal imbalances, high temperatures, stress and debilitating diseases. In conclusion, IG diagnosis necessitates semen analysis, testicular biopsy or fine-needle aspiration, and FSH testing, whereas IC diagnosis requires preputial and penile examinations. Diagnostic aids include serum and seminal biomarkers.

Azoospermia in male dromedary: Clinical findings, testicular biopsy, serum follicle stimulating hormone and seminal biomarkers.

Azoospermia is not an uncommon infertility problem in the male dromedary (Camelus dromedarius). Azoospermia was investigated via clinical findings, testicular biopsy as well as the evaluation of follicle stimulating hormone (FSH), concentration of camel testis protein (TEX101) and camel epididymis-specific extracellular matrix protein (ECM1) in seminal fluids. Azoospermic male camels (AZOO group, n = 28) that had been detected to be infertile as a result of lack of resulting pregnancies after repeated mating's for at least one season were included in this study. Clinical examination, semen analysis and testicular biopsy sampling were conducted for each individual animal. Blood samples were collected from the AZOO and from reference fertile males (FERT group, n = 8) for the assay of FSH hormone and semen biomarkers (TEX101 and ECM1). There were bilaterally normal-sized testes in 42.8%, bilaterally small-sized testes in 35.7%, bilaterally large-sized testes in 7.1%, no testicles in 7.1% and only one testicle in 7.1% of azoospermic animals. Sertoli cell-only syndrome (SCO) and maturation arrest were observed in 78.6% and 21.4% of the animals, respectively. There were greater concentrations of FSH in the AZOO group compared with the FERT group (P = 0.01). In conclusion, azoospermia in dromedary camels is mainly associated with spermatogenic defects and greater serum FSH concentrations. Seminal biomarkers, therefore, might be feasible indicators for identifying azoospermia in the male dromedary camels and the condition of non-obstructive azoospermia was seemingly prevalent in the male dromedary camels in the present study.

Reference values for the genitalia of male dromedary before and after puberty using caliper and ultrasonography in subtropics.

The aim of the present study was to characterize the dynamic changes of the testes and the pelvic genitalia in vivo in male dromedary. Eighty-one clinically healthy male dromedaries aged 1.5 to 12 years were assigned for the present study. Testicular length, breadth, and depth as well as epididymal head and tail were measured using caliper and ultrasonography. The pelvic genitalia, including bulbourethral gland, prostate, and pelvic urethra were examined using ultrasonography. The results revealed that the three dimensions of the testes and epididymal tail and head showed significant increase with age (P < 0.01). Concerning the epididymal measurements, differences between the pre- and peri-pubertal groups were not significant. Left testes tended to be larger than the right (not statistically significant) although only the breadth of the left testes in the prepubertal group was significantly larger (P < 0.05). The volume of both testes correlated positively with the age (r(2) = 0.91 for left and 1.00 for the right, P < 0.01). There were no significant correlations between the values measured using caliper and those by ultrasonography between groups, but the correlation was highly significant (P < 0.01) for the total number of the examined animals. There were significant and steady increases of the size of bulbourethral gland in all examined groups (P < 0.01). Pars disseminata of the prostate gland and pelvic urethra were significantly higher in sexually mature compared with prepubertal groups (P < 0.01). It was concluded that ultrasonography is a useful tool in studying the developmental changes of the testes and accessory glands of the male dromedary. The obtained data could provide a reference values for predicting camel puberty and future fertility.

Insulin-like growth factor-I as a follicular growth promoter during early pregnancy in thoroughbred mares.

To elucidate the physiological role of insulin-like growth factor-I (IGF-I) during early pregnancy in mares, number of ovarian follicles was monitored ultrasonically during different stages of the first trimester of pregnancy in 36 thoroughbred mares. From 9 of 36 mares, blood samples were collected weekly from the mating day till the end of the first trimester of pregnancy and plasma IGF-I profiles were examined with other hormones, like follicle stimulating hormone (FSH), luteinizing hormone (LH), ir-inhibin, progesterone and estradiol-17beta. Plasma IGF-I level fluctuated throughout the studied period with four peaks on the 7th, 28th, 49th and 84th days of pregnancy. Plasma IGF-I showed a positive correlation with plasma FSH (P<0.05), whereas no correlation was found with other hormones during the studied period. Plasma IGF-I had no correlation with the foetal size, but positive correlation with the number of large (> 30 mm) and medium (10-30 mm) follicles. These results suggested that IGF-I might produce from the medium and large follicles during early pregnancy and promote to develop their growth via pituitary FSH mediated effects in the mares.

Effect of active immunization of pony mares against recombinant porcine inhibin alpha subunit on ovarian follicular development and plasma steroids and gonadotropins.

Two pony mares were immunized against recombinant porcine inhibin alpha subunit three times with 39 day intervals. Clinical findings and endocrinological changes before immunization were taken as the control. The first significant rise in the anti-inhibin titre (P<0.05) in the circulation was found 27 days after the first injection. Maximum binding activity was reached by the 12th day after the second booster dose. The number of small, medium and large sized follicles had increased significantly compared to before immunization (11.75 +/- 4.30, 2.75 +/- 0.69 and 2.51 +/- 0.63 vs 6.50 +/- 1.43, 1.83 +/- 0.44 and 1.33 +/- 0.38, respectively), but the ovulation rate remained unchanged after immunization. The average plasma concentration of FSH and estradiol-17beta during the estrous cycle increased significantly (P<0.05) after immunization. These results suggest that immunization against inhibin is a useful tool to increase the number of ovarian follicles during the estrous cycle of pony mares. Moreover, the present study supported the concept that inhibin plays a major role in the control of follicular growth through its inhibitory effect on FSH secretion synergistically with steroid hormones.

Endocrinological changes before and after removal of the granulosa theca cell tumor (GTCT) affected ovary in 6 mares.

To clarify the endocrinological characteristics of the mares with granulosa theca cell tumor (GTCT), peripheral plasma samples from the 6 mares affected with GTCT were collected before and after the surgical removal of the affected ovary. Concentrations of testosterone (T), follicle stimulating hormone (FSH), luteinizing hormone (LH), immunoreactive-inhibin (ir-INH), progesterone (P) and estradiol-17beta (E(2)) in the plasma samples were measured by radioimmunoassay. Before removal of GTCT in all cases, the concentrations of T were significantly higher than those of normal mares at the breeding and non-breeding seasons, whereas plasma concentrations of FSH, LH, ir-INH, P and E(2) were lower. After surgical removal of the affected ovary, the circulatory concentrations of T was declined, but the concentrations of other hormones were constantly low as compared with those of normal mares. The present study suggests that 1) the source of higher T may be due to the abnormal follicles in ovary of GTCT, 2) in the case of GTCT the elevated level of T is observed due to the lack of aromatase, and 3) the high level of T is a typical characteristics for GTCT in mares. It is also suggested 4) due to the elevated levels of T the concentrations of gonadotropins may be suppressed.

Localization of inhibin alpha-, betaA- and betaB-subunits and aromatase in ovarian follicles with granulosa theca cell tumor (GTCT) in 6 mares.

To clarify the morphological and immunohistochemical characteristics in mares with granulosa theca cell tumor (GTCT), the localization of inhibin subunits (alpha, betaA, betaB) and aromatase in the granulosa cell layers and theca layers in the ovarian follicles were determined by immunohistochemical staining. The follicles were obtained from the ovaries of 6 mares with GTCT and 4 normal mares as controls. Immunohistochemically, inhibin alpha-subunit was localized in the granulosa cells of all follicles showing different sizes in all GTCT cases and betaA- subunit was localized in two GTCT cases in all sized follicles. But inhibin betaB- subunit and aromatase were not localized in GTCT cases. On the other hand, inhibin alpha-, betaA-, and betaB-subunits and aromatase were localized in the large and medium sized follicles, but inhibin betaA- and betaB-subunits and aromatase were not stained in the small sized follicles in normal cases. These findings suggest that some mares with GTCT can secrete dimeric inhibin (inhibin A), but all GTCT cases cannot secrete inhibin B. By the results of aromatase staining it is clear that testosterone is not converted into estradiol due to the lack of aromatase in the GTCT follicles.

Spontaneous repair of the atrophic contralateral ovary without ovariectomy in the case of a granulosa theca cell tumor (GTCT) affected mare.

A 21 year old thoroughbred mare with granulosa theca cell tumor (GTCT) in the right side and atrophic contralateral ovary was investigated in this study. After arrival at our laboratory on 10th December 1999, the clinical diagnosis of GTCT was examined by rectal palpation and ultrasonographic image of ovaries. Plasma from peripheral blood was collected in the breeding and non-breeding seasons for hormonal analysis. The results showed that the contralateral ovary regained normal activity without any treatment of the GTCT affected ovary and contained follicles showing different sizes 19 months later. However, the affected right ovary, which became smaller after 4 months, was totally inactive without any follicle. The observations clearly demonstrate that without any treatment of the GTCT affected ovary, a mare can return to her normal estrous cycle within a certain period in some GTCT cases.