Ultrasonographic and morphological sequelae to repeated testicular biopsies in stallions in relation to the lateral branching of the testicular artery

The objectives of this study were to investigate the sequelae to repeated testicular biopsies in stallions and to determine if arterial injuries can be prevented. This study was part of a larger project focused on the antispermatogenic effects of an oral contraceptive compound, RTI‐4587‐073(l), which was given to 3 mature Miniature horse stallions, while another 3 received a placebo treatment. Testicular biopsies were taken once before treatment and 3 times after treatment. They were obtained from alternating testes, 2 procedures per testis, 2 samples each time, using a 18 gauge split‐needle core biopsy instrument (penetration depth: 22 mm). Colour Doppler ultrasonography was performed prior to the procedure to detect and thus avoid lateral branches of the testicular artery. Testicular parenchyma was evaluated ultrasonographically just before, and 2–6 h, 3 days and weekly after the biopsies were obtained. Changes in testicular volumes and semen parameters were monitored. All stallions were then castrated and the testes were evaluated for any gross pathology. There was no major scrotal swelling or gross haemorrhage after the procedures. Only mild, focal lesions were found in the testicular parenchyma 3 and 7 days after taking the biopsies. Three lateral branches of the testicular arteries were punctured, but there was no evidence of significant bleeding or other complications associated with these injuries. We conclude that repeated testicular biopsies may be taken from stallions without causing major complications, regardless of a presence or absence of the lateral branches of the testicular artery. Furthermore, we conclude that using colour Doppler ultrasonography to detect the lateral branches of the testicular arteries is unreliable; however, puncturing these vessels during a biopsy procedure does not necessarily result in significant haemorrhage.     

Ultrasonographic evaluation of induced testicular lesions in male goats

To study testicular ultrasonographic features, unilateral testicular artery ligation was performed in 12 bucks to induce predictable ischemic necrosis of the testis. Both scrotal testes then were scanned subsequently on days 1, 3, 5, 7, 15, and 30. On each of the designated days, 2 bucks each were castrated and the excised testes were rescanned outside the scrotum before gross and histologic examinations. In each goat, the testis without the ligated artery served as a control. Gross and histopathologic findings in the testis were compared with ultrasonographic observations. As the study progressed from day 0 to day 15, visually and microscopically, the testis involved in the testicular artery ligation had edema that changed to areas of peripheral necrosis with thickening of the testicular capsule. On day 30, the affected testis had granulation tissue in the areas that previously had been necrotic. Ultrasonographically, the testicular changes were consistent with the gross and histopathologic findings. There were areas of decreased echogenicity that corresponded to the necrotic areas and foci of high echogenicity that corresponded to areas of tubular mineralization. On day 30, the granulation tissue was difficult to differentiate ultrasonographically from normal testicular parenchyma. Between days 3 and 15, hydrocele was seen ultrasonographically in several goats. The excellent association between ultrasonographic findings and gross and histologic changes suggested that testicular ultrasonography might be a valuable diagnostic tool for assessing testicular disease in domestic animals.     

Sampling intensities and replication requirements for detection of treatment effects on testicular function in bulls and stallions: a statistical assessment

Data from testes of 16, 2- to 3-yr-old stallions and 34 yearling beef bulls were utilized in a components of variance approach to calculate the number of observations required per testis and(or) the number of animals required per treatment group to provide experiments of known sensitivity and precision, where treatment was to be assessed by one of several endpoints. The latter included paired testes weight, seminiferous tubular diameter, the number of germ cells per seminiferous tubular cross-section, or the number of elongated spermatids per gram of testicular parenchyma or per testis. For all variables for which several observations were available for each testis, precise assessment of any given male required far fewer observations than have been used routinely. However, replication requirements varied substantially in relation to the sensitivity (i.e., size of difference to be detected) and precision (i.e., Type I and II error probabilities) desired. Replication requirements were greater for stallions than for bulls, particularly at higher levels of sensitivity, for which requirements for both species were very large. The data presented should permit future experiments involving assessment of these endpoints to be designed with known sensitivity and precision and with optimal efficiency and cost-effectiveness.     

Bilateral Testicular Mixed Germ Cell-sex Cord-stromal Tumours in a Stallion

An 18-year-old Friesian stallion was examined approximately one week after reportedly presenting scrotal swelling due to torsion of the spermatic cords. Upon presentation no scrotal swelling was noted, the testes were normally oriented, and no abnormalities of the spermatic cords were noted. However, both testes were smaller than expected for a mature stallion and deep palpation revealed that the consistency of the testes was nodular. Ultrasonographic evaluation of the testes revealed diffuse heterogeneous parenchyma with multiple hypoechoic nodular areas. Grossly, the testicular parenchyma was effaced by multiple gray-tan nodules of varying consistency interspersed with gray-white bands of tissue. Microscopic analysis revealed multiple pleomorphic neoplastic foci disseminated throughout both testes. Histological and immunohistochemical features were atypical and consistent with the diagnosis of bilateral testicular mixed germ cell-sex cord stromal tumours. Bilateral testicular tumours and testicular mixed cell tumours are extremely rare in stallions and this is the first report of bilateral testicular mixed germ cell-sex cord-stromal tumours in a stallion. Our findings indicate that certain ultrasonographic characteristics are suggestive of testicular tumour and that immunohistochemistry markers can be used to better characterize testicular neoplasms in stallions.     

Lateral Branches of the Testicular Artery Affect Testicular Shape in Adult Stallions

The objective of this study was to investigate the arterial patterns of the stallion testis in relation to testicular shape. Two hundred and fifty-one stallion testes were evaluated for the presence of the lateral branches of the testicular artery. Seven specimens had their testicular arteries filled with latex milk, fixed in 70% alcohol, and dissected. Two hundred six specimens (82%) had a single testicular artery and no lateral branches; 39 testes (16%) had one lateral branch of the testicular artery; and six testes (2%) had two lateral branches of the testicular artery each. The lateral branches of the testicular artery obtained from the adult stallions, more than 5 years old, were associated with distinct lateral bulging, giving them a pear-like shape, whereas similar vascular pattern in young colts, less than 1 year old, did not cause similar shape change. Five distinct patterns of the branching of the testicular artery were determined. We concluded that the lateral branches of the testicular artery are present in approximately 20% of stallion testes. This anatomic pattern is associated with a lateral bulge that develops slowly over several years and is associated with a change in testicular shape from an ellipsoid in colts to a pear-like shape in adult stallions.     

The complex blood supply to the equine testis as a cause of failure in laparoscopic castration

REASONS FOR PERFORMING STUDY: Intra-abdominal ligation/ transection of the spermatic cord may result in necrosis of the testis; castration of abdominal cryptorchids via laparoscopy has therefore become common. Notwithstanding some adaptations of the technique, a small percentage of operations fail, prompting research into the anatomical background and clinical relevance of the procedure. HYPOTHESIS: That an alternate blood supply may prevent complete necrosis of the testis after spermatic cord transection. OBJECTIVE: To establish the prevalence of the problem in normal and cryptorchid stallions. METHODS: In a preliminary study, the spermatic cords of 8 normal stallions were ligated and transected at different sites and in various manners. Five weeks later the testes were removed and the vitality of both the testes and epididymes was evaluated. In a prospective clinical trial, intra-abdominal spermatic cord transection was performed in 241 cryptorchid and normal stallions. In cases of surgical failure, the testes were removed and histology performed. RESULTS: Examination of the specimen removed from the 8 animals of the preliminary study revealed that all epididymes were completely or largely spared. All except one testis were completely necrotic. In the patients that underwent surgery all abdominally retained testes (n = 123) were necrotic, while 5 out of 88 inguinally retained and 8 out of 236 normally descended testes had partially survived. The pattern of survival differed between inguinally retained and normally descended testes. The epididymes of these 13 horses were (largely) vital. The (partial) survival of the epididymes and inguinally retained testes was ascribed to an alternate blood supply via anastomosing vessels derived from the cremasteric artery. A tributary from the external pudendal artery was considered responsible for the partial survival of normally descended testes. CONCLUSIONS AND POTENTIAL RELEVANCE: After intra-abdominal transection of the entire spermatic cord, 5.6% of inguinally retained and 3.4% of normally descended testes failed to become completely necrotic, as a result of an alternate blood supply via the cremasteric and/or external pudendal artery. Therefore, laparoscopic castration without orchidectomy cannot be recommended as a trustworthy method for castration of inguinal cryptorchids and normal stallions.     

Effects of Hemicastration on Testes and Testosterone Concentration in Stallions

The endocrine system is critical to the maintenance of testicular function. The homeostasis of sex hormone levels is orchestrated by positive and negative feedback systems controlled by the hypothalamic-pituitary-gonadal axis. This study investigated the long-term effects of hemicastration on testicular size and function in stallions. Four Thoroughbred stallions, 4–6 years of age, were included in this study. Several parameters, including testicular weight and volume, plasma testosterone concentrations, VASA-positive germ cell populations and cross-sectional areas of the seminiferous tubules were compared in stallions that underwent two hemicastrations, approximately 11 months apart. The weights and volumes of testes harvested at the second hemicastration were significantly higher than those of testes collected at the first hemicastration. However, VASA-positive germ cell populations and the cross-sectional areas of seminiferous tubules were not significantly different between testes harvested at the first and second hemicastrations. Similarly, plasma testosterone concentrations measured weekly for 3 weeks before the first hemicastration, 3 weeks after the first hemicastration, and 3 weeks before the second hemicastration were not significantly different. Our results suggest that hemicastration results in compensatory enlargement of the remaining testis and compensatory steroidogenesis to maintain normal reproductive function in stallions.     

Function of contralateral testis after artificial unilateral cryptorchidism in dogs

The effects of a cryptorchid testis on the contralateral testis were investigated after artificially producing unilateral cryptorchidism in 8 beagle dogs. Bilateral testicular biopsy and collection of spermatic vein blood and peripheral vein blood were performed at the time of the operation to produce the cryptorchidism and 52 weeks later. The testicular tissue was used for histological examination by light microscopy and measurement of the testicular transferrin (Tf) concentration by enzyme immunoassay. Plasma testosterone (T), estradiol-17 beta (E2), and luteinizing hormone (LH) levels were measured by radioimmunoassay. Semen was collected weekly and its quality was examined. No spermatogenesis was observed in the cryptorchid testes at 52 weeks after the operation, and the number of germ cells in the contralateral testes had decreased but the number of Sertoli cells did not change. The Tf concentration in both testes had also decreased. The mean total number of sperm between 48 and 52 weeks after the operation (194 x 10(6)) was less than half the number before the operation (510 x 10(6)). Mean spermatic vein plasma T levels (51 ng/ml) in the cryptorchid testes 52 weeks after the cryptorchid operation were significantly lower than before the operation (91 ng/ml; P < 0.05). By contrast, spermatic vein plasma E2 levels (80 pg/ml) were significantly higher than the values before the operation (51 pg/ml P < 0.05). The peripheral plasma LH levels decreased. These findings indicate that a large quantity of E2 secreted by the cryptorchid testis inhibits the endocrine and spermatogenic functions of the contralateral testis in the dog. In particular, it is assumed that dysfunction of the contralateral testis is associated with Sertoli cell dysfunction suggested by the low Tf concentration.     

Testicular seminoma associated with torsion of the spermatic cord in two cryptorchid stallions

Two adult horses had colic attributable to spermatic cord torsion and strangulation of abdominally retained neoplastic testes. Both horses had caudal abdominal soft tissue masses palpable per rectum. One horse was treated successfully by surgical removal of the testis, and the other was euthanatized without treatment. Histopathologic diagnosis of the involved testes was testicular seminoma. Spermatic cord torsion of an abdominally retained testis should be considered in the differential diagnosis of signs of abdominal pain in cryptorchid stallions, especially those with a palpable caudal abdominal mass.     

Acute and Chronic Effects of a Contraceptive Compound RTI‐4587‐073(l) on Testicular Histology and Endocrine Function in Miniature Horse Stallions

The objective of this study was to evaluate acute endocrine effects as well as histological changes in testicular parenchyma induced by the contraceptive compound RTI‐4587‐073(l). Six miniature stallions were used in this experiment. The treatment group (n = 3) received one oral dose of 12.5 mg/kg of RTI‐4587‐073(l), and the control group (n = 3) received placebo only. The stallions' baseline parameters (semen, testicular dimensions, endocrine values) were collected and recorded for 5 weeks before treatment and for 6 weeks after treatment. Multiple blood samples were collected for endocrine analysis. Testicular biopsies were obtained before treatment, 1 day after treatment and every other week after treatment. Ultrasound exams were performed to monitor the dimensions of the stallions' testes. All stallions were castrated 6 weeks after treatment. Sperm numbers, motility and percentage of morphologically normal sperm decreased (p < 0.05), while the number of immature germ cells increased in ejaculates from treated animals (p < 0.05). Serum concentrations of inhibin and follicle‐stimulating hormone did not change. Testosterone concentrations initially transiently decreased (p < 0.05) after administration of RTI‐4587‐073(l), and increased several days later (p < 0.05). Testicular content of testosterone and estradiol 17‐β was lower in treated stallions than in control stallions on Day 1 after treatment (p < 0.05). Severe disorganization of the seminiferous tubules, significant loss of immature germ cells and complete depletion of elongated spermatids were observed in testicular biopsies obtained from treated stallions 1 day, 2 and 4 weeks after treatment. These changes were still present in the testicular samples taken from treated stallions after castration. The results of this study confirmed that RTI‐4587‐073(l) has antispermatogenic effects in stallions. Furthermore, we concluded that this compound causes acute sloughing of immature germ cells from the seminiferous tubules. RTI‐4587‐073(l) has significant but transient effects on Leydig cell function in stallions.     

How to Perform and Interpret Testicular Fine Needle Aspiration in Stallions

Although several methods of testicular biopsy have been proposed previously, testicular fine needle aspiration (FNA) has proved to be the simplest, the most rapid, inexpensive, and overall the least invasive technique for obtaining testicular biopsies. Testicular FNA is indicated for fertility investigations in stallions with oligozoospermia or azoospermia. It is also used for differential diagnosis of testicular enlargement. After sedation, the stallion’s testis is punctured to obtain testicular parenchyma samples containing cells mainly from the seminiferous epithelium. The material obtained is used to perform smears which are analyzed for identification and quantification of germ cells and Sertoli cells. The results are based on the presence of the cell types found in the smears and the proportions of Sertoli cells per germ cells. In addition to being a very useful diagnostic tool, testicular FNA is also used for follow-up examinations, as it is minimally invasive.     

Postnatal testicular development and actin appearance in the seminiferous epithelium of the Habu,Trimeresurus flavoviridis

The postnatal testicular development and actin distribution in the seminiferous epithelium were examined by light microscopy, using the testes of the Habu (Trimeresurus flavoviridis; snake) from 0-year-old to 3-year-old. At 0-year-old (about 1 month after birth), the testis was quite small in size, and the seminiferous epithelium was composed of only Sertoli cells and large spermatogonia. Actin immunoreactivity was observed in the peritubular myoid cells, but could not be detected in the seminiferous epithelium. At 1-year-old (about 10 months after birth), the testicular size increased to a great degree. In the seminiferous epithelium, spermatocytes newly appeared. Actin could still not be detected in the seminiferous epithelium. At 2-year-old (about 1 year and 10 months after birth), the testes continued to develop in size. In the seminiferous epithelium, elongate spermatids and round spermatids were frequently seen, in addition to Sertoli cells, spermatogonia and spermatocytes. Thus, active spermatogenesis was clearly recognized at this age. Moreover, the actin distribution in the seminiferous epithelium was observed at the site between Sertoli cells and spermatids, as well as that at adult stage. The immunoreactivity of actin in the peritubular myoid cells gradually increased from 0-year-old to 2-year-old. Conclusively, it seems likely that spermatogenesis in the Habu initiates at 2-year-old, accompanying with the appearance of actin in the seminiferous epithelium.     

VASA (DDX4) is a Putative Marker for Spermatogonia,Spermatocytes and Round Spermatids in Stallions

Expression of the protein DDX4/MVH, or VASA, has been reported in germ cells of several species. The main objectives of this study were to (i) investigate VASA expression patterns in testicular cells of stallions at two different reproductive stages (pre‐pubertal and post‐pubertal) and (ii) evaluate the use of VASA antibody as a molecular marker for single germ cells from stallions. Testicular tissues were obtained from stallions and categorized as pre‐pubertal and post‐pubertal based on the formation of lumen and status of spermatogenesis on the cross section of seminiferous tubules. The results of Western blot showed a VASA protein band located at 76 kDa, indicating that the rabbit antibody has a cross‐reactivity with horse testicular tissues. The result of immunolabelling showed that VASA was expressed in the cytoplasm of spermatogonia at both reproductive stages and in spermatocytes and round spermatids at the post‐pubertal stage. GATA4‐positive Sertoli cells and Leydig cells located in the interstitial space were not immunolabelled with VASA. These results suggest that VASA can be utilized as a molecular marker for germ cells of stallions at pre‐pubertal and post‐pubertal stages. Interestingly, immunolabelling intensity was significantly higher in pachytene spermatocytes compared to spermatogonia and round spermatid. VASA antibody was also effective for staining of single germ cell preparations. In conclusion, VASA protein expression can be used as a marker for identification of spermatogonia, spermatocytes and round spermatids in testicular tissues of stallions.     

Effects of Intratesticular Injection of 70% Glycerin on Stallions

The removal of endogenous germ cells of recipient stallions is a key step to produce donor germ cell-derived sperm using the germ cell transplantation technique. Six Thoroughbred stallions were divided into a treatment (n = 3) and a control group (n = 3), and 70% glycerin (1, 2, 3-trihydroxypropane, 40 mL per testis) or phosphate-buffered saline, respectively, was locally injected into testes. General semen evaluation, libido, and testicular volume were performed weekly from 3 weeks before to 10 weeks after treatment. The number of round germ cells in the ejaculate was counted using a hemocytometer. The hematoxylin and eosin staining was performed on the cross sections of testicular tissue obtained 11th week of treatment. Plasma testosterone levels in blood collected weekly were measured using a colorimetric competitive enzyme immunoassay kit. The sperm number was significantly lower than that of the control group at 5 and 10 weeks after glycerin injection. No differences in the status of spermatogenesis in the cross sections of seminiferous tubules and testicular volume were found between the two groups. The 70% glycerin-treated stallions had reduced total and progressively motile sperm and exhibited a significantly higher population of round germ cells in the ejaculate. Testosterone levels, testicular volumes, and libido of stallions were not significantly different between the groups. In conclusion, although intratesticular injection of 70% glycerin may have caused disassociation of some germ cells in the seminiferous tubules for several weeks, it did not significantly ablate germ cells in the tubules at 11 week in stallions.     

2-Bromopropane induced germ cell apoptosis during spermatogenesis in male rat

2-Bromopropane (2-BP) causes testicular toxicity in humans and rats. However, the germ cell degeneration of testicular toxicity by 2-BP has not been understood. 2-BP at doses of 135, 405, and 1,355 mg/kg/day was daily injected subcutaneously into Sprague-Dawley rats for 28 days. At the dose of 1,355 mg/kg/day, 2-BP significantly decreased the weights of body and testes, eipididymis, seminal vesicle, and prostate, as well as daily sperm production. Atrophy of seminiferous tubules accompanied with degeneration of germ cells such as spermatogonia, spermatocytes, and elongated spermatids was observed in the testes of rats exposed to the 405 mg/kg/day and 1,355 mg/kg/day of 2-BP. TUNEL-positive germ cells were appeared in the 405 and 1,355 mg/kg/day of 2-BP-treated groups. In addition, ultrastructure alterations of apoptotic germ cells were observed by the electron microscopy study. Dead elongated spermatids were observed at 1,355 mg/kg/day after 28 days exposure. These results suggest that 2-BP impair spermatogenesis may result from apoptotic germ cell death.     

Stereological analysis of Wistar rat testis during early post-natal development

Quantitative analysis of testis structure was performed during early post-natal development in Wistar rats. For this purpose, at 1, 7, 14, 21, 28, 35, 42, 60 and 90 days after birth, weights and volumes of testes were recorded and fixed in Bouin's solution. For stereological studies, 5-μm paraffin sections were stained with haematoxylin-eosin. Relative and then absolute volumes of seminiferous tubules, germinal epithelium, seminiferous tubuli lumina and interstitial tissue were estimated by point-counting method. The results showed that weight and volume of testis in rats increase 75- and 86.28-fold during post-natal development, respectively. The greatest growth rate (2.96-fold) of testis was observed between days 35 and 42. Also, diameter of seminiferous tubules increased significantly (P<0.001) between different ages and represented 5.55-fold increase during post-natal development. The percentage volume of tubular tissue increased 1.4-fold between birth and 90 days of age. Interstitial tissue formed 36.68±0.90% of testicular parenchyma at birth. This percentage decreased progressively during post-natal development until 90 (9.00±0.55%) days of age. Lumen of seminiferous tubules was recognized at day 28, and its relative volume increased with age. This study provides systematic data on the stereological characteristics of developing Wistar rat testis.     

Impact of fine- or large-needle aspiration on canine testes: clinical, in vivo ultrasonographic and seminological assessment

The safety and consequences of fine- (FNA) and large-needle aspiration (LNA) to the testicular parenchyma and its normal function have not been thoroughly established. This study was performed to accurately assess, by serial clinical, in vivo ultrasonographic and seminological examinations, the type and extent of the effect of FNA or LNA on canine testes. Eighteen sexually mature, 1-2 years old, healthy laboratory Beagles were used. One of their testes was aspirated using a 23-G butterfly needle (FN) and the other using a 19-G butterfly needle (LN). Two dogs at a time were orchiectomized 10, 60 min, 2, 14, 29, 63, 76, 90 or 180 days post-aspiration. Five and 2 days and 1 h before aspiration (in all dogs), immediately post-aspiration, and 1, 2, 4, 7, 9, 14, 19, 29, 35, 43, 49, 56, 63, 70, 76, 90, 111, 132 and 180 days post-aspiration (in the remaining intact dogs), evaluation of scrotal surface temperature over each testis, evaluation of scrotum-testis volume by electronic sliding callipers, ultrasonographic evaluation of testicular volume and texture and clinical and semen examination were performed. Following FNA and LNA, the clinical and ultrasonographic appearance of the testis were normal. Sperm production nearly always remained unchanged, with the exception of a slight decrease in spermatozoal motility 2-14 days post-aspiration. However, even then, with the exception of six samples, spermatozoal motility was above normal values. Within the parameters of this experiment, testicular FNA and LNA have no ill effect on sperm production or clinical and ultrasonographic appearance of the canine testis, and therefore, both FNA and LNA should be considered safe.     

Sonographic characteristics of goat testis on water bath based ultrasonography

Assessing the health of the testes in domestic animals is an important aspect of the breeding soundness examination and selection. The aim of the present study was to develop a simple method for scanning and to establish ultrasonographically the gross anatomic structures of the goat testes. Six adult male goats were examined to study the sonographic appearance of normal testes and epididymides using a water bath based ultrasound scanning technique. The ultrasonographic examinations were done using a 5–9 MHz/60 mm (7.5 MHz) linear-array transducer and a B-mode scanner. The ultrasonographic examination was performed in goats after standardizing the procedure on six testes collected from slaughter house. Results showed that in live goats when the probe was placed directly over the scrotum it gave distorted and unclear image. In water bath method the entire scrotum was dipped into a container filled with water and linear probe was used to observe the sonographic features of the testis. Each testis was viewed vertically, resulting in longitudinal image which was frozen, measured and printed through a thermal printer. The results of the ultrasonogram revealed that the testicular parenchyma was homogenous and moderately echogenic throughout. The diameter (mean±se) of the right and left testes was 4.47±0.14 and 4.42±0.07 cm respectively and no significant difference was observed between the testes. The mediastinum testis was a 1.50±0.22 cm wide linear structure of greater echogenicity than the testicular parenchyma when viewed in the transverse plane and nearly circular echogenic “spot” in the midline of the testis when viewed horizontally. The head and tail of the epididymides were easily identified on all the testes, but the epididymal body and ductus deferens were difficult to identify consistently. The tail of the epididymis was easily identified on the distal end of the testis with sonolucent tubules and appeared sonographically as a ‘peaked cap’ upon the testicular parenchyma. The diameter (mean±se) of the tail of right and left epididymis was 2.11±0.18 and 1.92±0.06 cm and no significant difference was observed between epididymides. The vascular pampiniform plexus (1.42±0.18 cm) was easily identified on the proximal end of the testes. The tunics of the testes appeared as a bright echogenic line. Inter-testicular septum appeared between testes as a hyperechoic line. It is concluded that ultrasonography permits a noninvasive evaluation of the internal structure of the scrotum and testes and water bath based sonographic examination may prove to be a valuable simple diagnostic methodology for evaluating physiopathologic conditions of goat testes and can be employed as a routine investigative method during breeding soundness and clinical examination.     

Testosterone serum profile,semen characteristics and testicular biometry of Mangalarga Marchador stallions in a tropical environment

This study was conducted to characterize the daily profile of testosterone secretion and its mean concentrations in the four seasons as well as to evaluate the semen characteristics and testicular biometry of Mangalarga Marchador stallions throughout the year in a tropical region. Three stallions were submitted to semen collections and evaluation of testicular biometry every 14 days along a year. Blood samples were collected once at the middle of each season, in a 20‐min interval during 24 hr in order to evaluate the testosterone secretion profiles among seasons. Testosterone concentrations along the day were higher at the beginning of the afternoon (from 12:00 to 15:00 hr), but a circadian secretion was not clearly observed. Mean testosterone concentrations did not differ among seasons (p > .05), but a pattern of secretion along the day showed variations with higher concentrations in the afternoon during the winter. Ejaculate volume was higher during summer; however, sperm motility decreased in summer and spring. Total sperm in ejaculate, sperm morphology and testicular biometry kept constant along the year showing no differences among the seasons. The results demonstrated that in a tropical region, reproductive aspects of stallions did not show a clearly defined seasonal variation, and months of autumn and winter were not unsuitable for reproduction of the males.