Testicular Structure and Seminal Pathway in the Yellowtail Tetra Astyanax altiparanae (Characiforms: Characidae)

This study aimed to describe testicular and its main ducts structure in the yellowtail tetra Astyanax altiparanae, contributing to the knowledge of the region in which semen is produced, storage and released, focusing mainly on the dynamic of germinal epithelium and Sertoli cells during germ cell maturation. Ten sexually mature male A. altiparanae had their testes processed according to the routine protocols to optical microscopy. Moreover, spermatic ducts and tubular compartment of the testes of three specimens were perfused with vinyl resin for gross anatomy and scanning electron microscopy. Astyanax altiparanae testes are paired organs, separated for most of their extension, joining posteriorly in a spermatic duct formed by a squamous simple epithelium. Seminiferous compartment presents anastomosing tubular type organisation, and spermatogonia spread along its extent. Spermatogenesis is of cystic type, and there is no main testicular duct. Spermatogenesis develops in ‘waves’, from posterior to anterior part of the gonad. Thus, while sperm is storage posteriorly, spermatogenesis keeps maturing germ cells anteriorly, making the germinal epithelium very dynamic, holding Sertoli cells that change their function as a cystic envelope to produce secretions of the seminal fluid and store sperm. Such kind of development is thought to be responsible by the high prolificacy of this species.     

Influence of different warming temperatures on the vitrified testicular fragments from pre-pubertal cat

Testicular vitrification is an alternative to preserve the genetic material of pre-pubertal animals. However, there are few studies on post-vitrification warming. Hence, the aim was to compare the influence of different warming temperatures on vitrified testicular fragments from pre-pubertal cats. The testicles were fragmented and divided into a control group (non-vitrified) and vitrified, using an association between dimethylsulphoxide and glycerol. The vitrified fragments were warmed at 50, 55 and 60°C/5 s. Morphological and morphometric evaluations were carried out using classical histology. Afterwards, the mitochondrial activity was evaluated using Rhodamine 123. The data were expressed in mean and standard error. The differences were considered significant when p < .05. In the histomorphological analysis, the testicular fragment presented seminiferous tubules with poorly developed germinal epithelium, compatible with pre-pubertal animals. The group warmed at 50°C presented similar to the control regarding the maintenance of the integrity of the tubules and cells, without stromal rupture and lamina propria alteration, as well as regarding the maintenance of the junctions between the cells. The group warmed at 55°C showed reduction of the cell junctions, and the one warmed at 60°C had increased detachment of the basement membrane (p < .05). The warming caused a reduction in the tubular diameter inversely proportional and progressive to the increase in temperature, with the highest diameter in the control group and the lowest in the 60°C group. The control group showed a lower incidence of Rhodamine 123, followed in ascending order of the warmings at 55 and 60°C. The higher mitochondrial activity was obtained with 50°C, showing an increase of the metabolic cell function at this temperature. It was concluded that the testicular fragment of pre-pubertal cats presents a better preserved morphology, morphometry and viability when warmed at 50°C.     

The Correlation between Age,Body Weight and Testicular Parameters in Murrah Buffalo Bulls Raised in Brazil

Buffalo are an economically important source for meat and milk production, especially in Brazil. However, important aspects of their biology remain unknown thus far. Herein, we describe the reproductive characteristics of male Murrah buffalo (Bubalus bubalis) raised under extensive management conditions by applying biometrics associated with testicular weight. We analyzed seven males, divided into two groups: G1, which consisted of four 18-month-old animals, and G2, which consisted of three 24-month-old animals. Testicular development occurs slowly in Murrah buffalo, suggesting a delay of sexual maturity. The biometric testicular parameters analyzed were scrotal circumference, testicular weight, testicular length, testicular width, testicular thickness and testicular circumference. Our data indicate strong correlations between SC, age and body weight, and additional significant relationships were identified between body weight, age and other testicular parameters. Thus, these parameters are suitable indicators when selecting bulls for breeding purposes.     

Trehalose protects testicular tissue of dairy goat upon cryopreservation

The aim of this study was to investigate whether the addition of trehalose to cryomedia reduces cellular damage and improves gene expression in cryopreserved dairy goat testicular tissues. Testicular tissues were cryopreserved in cryomedia without or with trehalose at a concentration of 5%, 10%, 15%, 20% or 25%. Cryopreserved testicular tissues were analysed for TUNEL‐positive cell number, expression of BAX, BCL‐2, CREM, BOULE and HSP70‐2. Isolated Leydig cells from cryopreserved tissue were cultured, and spent medium was evaluated for testosterone level. The results showed that though the TUNEL‐positive cell number increased in cryopreserved testicular tissues, the presence of trehalose reduced apoptotic cell number significantly. Quantitative real‐time polymerase chain reaction results showed that although the expression of BAX was upregulated following cryopreservation, the presence of trehalose downregulates it in cryopreserved testicular tissues. Expression of BCL‐2, CREM, BOULE and HSP70‐2 was downregulated following cryopreservation but the presence of trehalose significantly upregulated their expression in cryopreserved testicular tissues. Leydig cells isolated from testicular tissues cryopreserved with trehalose produced higher testosterone than the one without it (control). These results suggest that trehalose has a protective role in cryopreservation of dairy goat testicular tissue, and the most suitable trehalose concentration for cryopreservation is 15%.     

Post mortem studies on infertile buffalo bulls: testicular histology

Testicular tissues of 22 buffalo bulls (Bubalus bubalis) which suffered from three types of infertility were examined histologically. Nine bulls with no sexual libido showed underdeveloped seminiferous tubules; five of them also had various forms of germinal tissue hypoplasia (bilaterally complete, partial or incomplete) and in the other four the seminiferous tubules showed developed layers of germinal epithelium but no complete spermiogenesis. Among 11 bulls which had always produced poor quality semen one suffered from incomplete bilateral testicular hypoplasia and two had simple testicular degeneration; five showed marked testicular degeneration associated with fibrosis of intertubular spaces, and in three there was intertubular fibrosis and tubular stasis. In two bulls in which the quality of semen had deteriorated one showed bilateral partial testicular hypoplasia and the other had bilateral testicular degeneration with unilateral intertubular fibrosis and tubular stasis.     

Prevalence of the AMHR2 mutation in Miniature Schnauzers and genetic investigation of a Belgian Malinois with persistent Müllerian duct syndrome

Persistent Müllerian duct syndrome (PMDS) is a sex‐limited disorder in which males develop portions of the female reproductive tract. Important consequences of PMDS are cryptorchidism and its sequelae of infertility and increased risk of testicular cancer. Anti‐Müllerian hormone (AMH) and its receptor (AMHR2) induce the regression of the Müllerian ducts in male embryos. In Miniature Schnauzer dogs, the genetic basis has been identified as an autosomal recessive nonsense mutation in AMHR2, but the allele frequency of the mutation is unknown. Thus, the primary objective of this study was to estimate the prevalence of the AMHR2 mutation in North American Miniature Schnauzers, in order to ascertain the value of genetic testing in this breed. An additional objective was to determine whether mutations in AMH or AMHR2 were responsible for PMDS in a Belgian Malinois; this would aid development of a genetic test for the Belgian Malinois breed. Genomic DNA from 216 Miniature Schnauzers (including one known PMDS case) was genotyped for the AMHR2 mutation, and DNA from a single PMDS‐affected Belgian Malinois was sequenced for all coding exons of AMH and AMHR2. The Miniature Schnauzer cohort had an AMHR2 mutation allele frequency of 0.16 and a carrier genotypic frequency of 0.27. The genetic basis for PMDS in the Belgian Malinois was not determined, as no coding or splicing mutations were identified in either AMH or AMHR2. These findings support a benefit to AMHR2 mutation testing Miniature Schnauzers used for breeding or with cryptorchidism.     

Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

Evaluation of the Breeding Soundness of Male Camels (Camelus dromedarius) via Clinical Examination,Semen Analysis,Ultrasonography and Testicular Biopsy: A Summary of 80 Clinical Cases

Male camel infertility is a heterogeneous disorder. A variety of factors may adversely affect sperm production and function and impair fertility. This study was designed to evaluate the sensitivity and specificity of ultrasonography and testicular biopsy in the evaluation of the breeding soundness of male dromedaries compared with results obtained by clinical examination and semen analysis. Eighty‐four male dromedary camels (5–15 years old) were used in this study during the rutting season (November–May). Four sexually mature male camels were used as controls. These animals were apparently healthy and had histories of normal fertility. Eighty infertile male camels were subjected to an algorithmic approach based on information collected during careful examinations of the camels' breeding histories, clinical examinations, testicular evaluations, testicular ultrasonographies, the results of the semen analyses and testicular biopsies to diagnose the camels' infertilities. The differences in the semen parameters between the control and infertile male camels were highly significant (p < 0.01). Regarding the diagnoses of male camel infertility, the results of testicular ultrasonographies and biopsies were compared with those from the semen analyses, and the accuracies of these tests were 92.5% and 90%, respectively. Additionally, the results of the testicular ultrasonographies were matched with those of the testicular biopsies of the infertile animals, and this comparison resulted in 85% accuracy. Testicular biopsy is a promising method that, along with a carefully performed history, clinical examination, an appropriate testicular ultrasonography procedure and semen analysis, can afford veterinarians the opportunity for more precise diagnosis and treatment of many dromedary infertility disorders.     

Testicular Damage from Anabolic Treatments with the β2-Adrenergic Agonist Clenbuterol in Pigs: A Light and Electron Microscope Study

The morphological consequences of anabolic clenbuterol treatment on the testicular parenchyma were investigated in 30 pigs at morphological and ultrastructural levels. Clenbuterol was given with food (1 ppm). In the first group (n=10), treatment was maintained until slaughter (experimental period 3 months). In the second group (n=10), clenbuterol was withdrawn 2 weeks before slaughter (experimental period 2.5 months). A third group (n=10) of pigs not fed with clenbuterol served as controls. Animals were slaughtered at 9 months of age and samples of testicular parenchyma were collected for light and electron microscope studies. In the clenbuterol-treated groups, the interstitial cells showed a considerable increase in the organelles involved in testosterone production, with an increased development of the mitochondria, smooth endoplasmic reticulum, Golgi apparatus and lipid droplets compared to the control group. The seminal epithelium displayed many lipid vacuoles and evident signs of tubular involution, such as degenerating and multinucleate germ cells. Sertoli cells gave evidence of metabolic alterations such as large lipid deposits and cytolysosomes.     

Testicular feminization syndrome in a mare.

Testicular feminization syndrome was diagnosed in a mare with aggressive, stallion like behavior and a history of infertility. She was found to have a high baseline testosterone concentration suggesting that testicular tissue was present, and ovarian-like structures examined by use of transrectal ultrasonography had the appearance typical of testicular tissue. Although her external female genitalia appeared normal, her vagina ended in a blind sac, and no cervix or uterus were identified. Surgery was performed, and structures removed from the abdominal cavity were determined to be hypoplastic testicles. Removal of the testicular tissue resulted in complete resolution of her aggressive behavior. Chromosomal evaluation revealed that the mare had 64X,Y (normal male) karyotype. Testicular feminization syndrome is a condition characterized by insensitivity of reproductive tissues to androgens during development because of an abnormality in androgen receptors. This androgen insensitivity results in development of normal external female genitalia, with high testosterone concentrations being released from developing testicles. Testicular feminization syndrome has not been commonly diagnosed in horses, but should be considered as a differential diagnosis for overly aggressive mares with a history of infertility.     

Retracted: Gonad histology and serum 11‐KT profile during the annual reproductive cycle in sterlet sturgeon adult males,Acipenser ruthenus

The aim of this study was to assess monthly testicular development in the cultured breeding stock of sterlet, Acipenser ruthenus, using histological and serum sex steroid changes. Testicular development in the adult male was examined monthly and showed four distinct phases including resting, pre‐spawning, spawning and post‐spawning. Also, seasonal changes of the testes were described according to its variations in gonadosomatic index (GSI) during different phases of testicular development. Using histology, we identified continuous spermatogenesis and asynchronous gonad development pattern in the testes of male sterlet, which shows that regulation of annual gonadal cycle is influenced by season. Results also showed variation in the GSI value and number of spermatogenic cells according to each season during annual cycle of gonad, as the highest value of GSI was recorded during spawning phase (spring; March‐May). Hormonal profiles of 11‐ketotestosterone (11‐KT) showed peak, which indicated a seasonal pattern of gonadal development. The 11‐KT concentration increased considerably during the spermatogenesis (pre‐spawning phase) and remained quite high throughout the pre‐spermiation period. In the final phase of testicular development (spawning phase), the 11‐KT markedly dropped. This study undertook an examination of complete reproductive development in cultured sterlet sturgeon to provide a valuable guide for the future sterlet studies, and allows comparison of reproductive development between sturgeon species.     

Use of intralesional oestradiol concentration to identify a functional pulmonary metastasis of canine sertoli cell tumour

A seven-year-old, 31 kg male neutered Labrador was investigated for signs of feminisation syndrome and prostatic disease four years after castration and removal of a testicular sertoli cell tumour (SCT). Investigations revealed an elevated serum oestradiol-17β concentration, a pulmonary mass containing fluid high in oestradiol-17β and cystic changes in the prostate gland. The pulmonary mass was surgically excised and histologically confirmed to be a SCT metastasis. To the authors' knowledge, this is the first reported case of a proven functional extranodal SCT metastasis and the first to be diagnosed by oestradiol-17β measurement of intralesional fluid.     

The Annual Testicular Cycle of the Domestic Quail (Coturnix coturnix japonica)

A morphometric study was conducted on the testis of the domestic quail Coturnix coturnix japonica to determine testicular kinetics. We investigated the variability along the year of testicular parameters such as seminiferous tubule diameter, germinal epithelium height and amount of meiotic figures of maturing spermatids in the seminiferous epithelium and of sperm in the tubular lumen. The results of morphometric analysis showed the occurrence of an annual testicular cycle defined by four distinct phases: a resting phase (at the end of summer), a recrudescence phase (in the fall), a proliferative phase (at the end of winter and beginning of spring), and a regression phase (spring and summer). We also observed that the testes of adult quails present elevated and maximal spermatogenic activity in fall-winter (short-day period) and at the beginning of spring, respectively, and lower values in spring and summer (long-day periods), with minimum values at the end of summer.     

DNA methylation and expression of imprinted genes are associated with the viability of different sexual cloned buffaloes

The DNA methylation of imprinted genes is an important way to regulate epigenetic reprogramming of donor cells in somatic cell nuclear transfer (SCNT). However, the effects of sexual distinction on the DNA methylation of imprinted genes in cloned animals have seldom been reported. In this study, we analysed the DNA methylation status of three imprinted genes (Xist, IGF2 and H19) from liveborn cloned buffaloes (L group, three female and three male), stillborn cloned buffaloes (S group, three female and three male) and natural reproduction buffaloes (N group, three female and three male), using bisulphite sequencing polymerase chain reaction (BS‐PCR). The expression levels of these imprinted genes were also investigated by quantitative real‐time PCR (QRT‐PCR). The DNA methylation levels of H19 were not significantly different among the groups. However, the Xist in female and IGF2 in male of the S group were found to be significantly hypomethylated in comparison with the same sexual buffaloes in L group and N group (p < .05). Furthermore, the expression levels of Xist, IGF2 and H19 in the stillborn female cloned buffaloes of S group were significantly higher than that of the female buffaloes in the L group and N group (p < .05). The expression levels of IGF2 and H19 in the stillborn male cloned buffaloes in the S group were significantly higher than that of the male buffaloes in the L group and N group (p < .05). These results indicate that Xist may be associated with the viability of female cloned buffaloes, and IGF2 may also be related to the viability of male cloned buffaloes.     

Hepatitis B Surface Antigen S Gene is an Effective Carrier Molecule for Developing GnRH DNA Immunocastration Vaccine in Mice

Relatively molecular mass of GnRH antigens is small and hence needs to couple to a large carrier molecule to enhance its immunogenicity. This study investigated whether hepatitis B surface antigen S (HBsAg‐S) gene can be used as an effective carrier molecule for developing GnRH DNA immunocastration vaccine. Two copies of human GnRH gene were fused with HBsAg‐S gene for constructing a recombinant plasmid pVAX‐HBsAg‐S‐2GnRH that coded for 27 kDa target fusion protein. Ten male mice were divided into two equal groups, treatment and control. The vaccine (50 μg/mice) prepared in saline solution was injected into male mice at weeks 0, 1, 2, 4 and 7 of the experiment. Vaccine's efficacy was evaluated in terms of GnRH‐specific IgG antibody response, plasma testosterone levels, testicular weight and extent of the testicular tissue damage. The specific anti‐GnRH antibody titre in vaccinated animals was significantly higher than in controls in only 4th week of immunization (p < 0.05). In addition, vaccinated animals showed lower testicular weight than those of the controls (p < 0.05). Spermatogenesis in seminiferous tubules in vaccinated animals was suppressed. In conclusion, in this study, the engineered plasmid to be used as a GnRH DNA vaccine induced antibody response and suppressed spermatogenesis in mice. This suggests that HBsAg‐S gene can be an effective carrier molecule for developing GnRH DNA immunocastration vaccine when relatively molecular mass of the aimed antigens is small.     

Estimates of genetic correlations between testicular measurements and female reproductive traits in cattle

Data from 528 male and 645 female progeny of 63 sires were used to estimate genetic correlations between female and male reproductive traits. Data were from two Hereford herds involved in a long-term selection program of the North Carolina Agricultural Experiment Service. Testicular measurements of circumference, diameter, length and volume were obtained on bulls at 205 and 365 d. Testicular growth measures were defined as differences between 205-and 365-d measurements. Heifers were placed in the breeding herd as yearlings and given two breeding seasons to produce a calf. Traits utilized from females were three age-at-first-breeding traits, two age-at-first-calving traits, two pregnancy rate traits, rebreeding interval and calving interval. Genetic correlations were estimated from half-sib and from sire-daughter analyses. Seventy-five percent or more of the correlations of testicular measurements with pregnancy rats, age at first breeding and age at first calving were in the favorable direction. Average correlations were .62, -.55 and -.66, respectively. For each of the remaining female traits, approximately 50% of the correlations were favorable and the average correlations were small. Correlations were summarized by testicular measurement with favorable correlations given a negative sign. Testicular diameter had more favorable correlations (80%) than length, volume or circumference (70%). However, average correlations were similar (-.31, -.30, -.34 and -.26, respectively). Testicular measurements taken at either 205 or 365 d had the same percentage of favorable correlations (72%), while testicular growth measurements had a slightly higher percentage of favorable correlations (78%). Average correlations of 365-d measures were higher (-.38) than either 205-d or growth measures (-.25 and -.28, respectively). Heritabilities for testicular measurements tended to be moderate to high, while those for female reproduction tended to be low to moderate. These results suggest that selection for increased testicular size would lead to improvement in female reproduction, particularly an increase in calving rate and a decrease in age at first breeding.     

miR‐26a suppresses autophagy in swine Sertoli cells by targeting ULK2

A large number of microRNAs (miRNAs) have been detected from porcine testicular tissues thanks to the development of high‐throughput sequencing technology. However, the regulatory roles of most identified miRNAs in swine testicular development or spermatogenesis are poorly understood. In our previous study, ULK2 (uncoordinated‐51‐like kinase 2) was predicted as a target gene of miR‐26a. In this study, we aimed to investigate the role of miR‐26a in swine Sertoli cell autophagy. The relative expression of miR‐26a and ULK2 levels has a significant negative correlation (R² = .5964, p ≤ .01) in nine developmental stages of swine testicular tissue. Dual‐luciferase reporter assay results show that miR‐26a directly targets the 3′UTR of the ULK2 gene (position 618–624). In addition, both the mRNA and protein expression of ULK2 were downregulated by miR‐26a in swine Sertoli cells. These results indicate that miR‐26a targets the ULK2 gene and downregulates its expression in swine Sertoli cells. Based on the expression of marker genes (LC3, p62 and Beclin‐1), overexpression of miR‐26a or knock‐down of ULK2 inhibits swine Sertoli cell autophagy. Taken together, these findings demonstrate that miR‐26a suppresses autophagy in swine Sertoli cells by targeting ULK2.     

Evaluation of reproductive parameters in male Neotropical bats during dry and rainy months in a specific area of the Cerrado biome

The aim of this study was to analyse the reproductive aspects of male bats of three common species of the Phyllostomidae family: Artibeus lituratus, Platyrrhinus lineatus and Sturnira lilium, during dry and rainy months in a specific area of the Cerrado biome. Body weight was significantly higher during the dry months for S. lilium. The gonadosomatic index (GSI) and testicular weight were not significantly different between dry and rainy periods. The tubular parameters were significantly bigger in A. lituratus than in the other two species during both periods. No difference in the tubular/interstitial ratio was observed in any of the species during both periods. In both periods, all sperm cells and germ cell developmental stages were visible on seminiferous tubules whereas sperm cells were observed in epididymides of all sampled animals. The percentage of morphologically normal sperm was low (35%–60%), with no difference between periods. Spermatozoa from A. lituratus presented a leaf-shaped head, while the head was round-shaped in the other two species. In conclusion, our data suggest that males from the three studied species did not present reproductive latency during the most critical weather periods (dry and rainy months) in the metropolitan region of Brasilia, Brazil.     

Effect of active immunization against GnRH‐I on the reproductive function in cat

This study was designed to explore the effect of active immunization against maltose binding protein‐gonadotropin releasing hormone I hexamer (MBP‐GnRH‐I6) on the reproductive function in cats. Each immunized cat was administered twice intramuscularly in the neck at 16 and 20 weeks old. The concentrations of the testosterone and estradiol and the level of anti‐GnRH‐I antibody in the serum were measured by radioimmunoassay and ELISA, respectively. The results showed that the weight and size of testicles and ovaries, and the concentrations of serum testosterone and estradiol in the immunized animals were lower than those of the control cats (P < 0.05), but that the levels of anti‐GnRH‐I antibody were significant higher compared to control animals (P < 0.05). Testicular tissues from the immunized male cats showed that seminiferous tubules were depauperate with the lumen relatively empty and that the differentiation of spermatogonia was not obvious. Tissues from the immunized female cats showed that the ovaries had many primordial follicles and primary follicles, but no secondary follicle was observed. These results showed active immunization against MBP–GnRH‐I6 could make the gonads atrophy and reduce the concentrations of gonadal hormones, which suggested that MBP‐GnRH‐I6 was a very effective immunogen in the cat.     

Testicular shape,scrotal skin thickness and testicular artery blood flow changes in bulls of different ages

The aim of this study was to compare the biometric testicular characteristics, skin thickness and haemodynamics of the testicular artery of 12- and 24-month-old bulls using Doppler ultrasonography, the study was conducted using 48 indicus-taurus animals. The scrotal circumference (SC) and biometry characteristics of the bulls were measured to calculate the testicular volume. Doppler ultrasonography was used to obtain the haemodynamic values of the testicular artery. The skin thickness and volume were lower (p<.01) in the younger bulls (12 months:4.68 ± 0.68 mm; 168.76 ± 47.96 cm³) versus 24 months (5.05 ± 0.89; 499.73 ± 129.24 cm³) animals (p<.01). During diastole, mean velocity was lower in the 12 months (7.98 ± 3.83) than in the 24 months (11.37 ± 4.15) animals (p <.05). The 12-month-old animals had higher pulsatility and resistivity indices (0.49 ± 0.02; 0.51 ± 0.20) compared to the 24-month-old animals (0.32 ± 0.16; 0.40 ± 0.15) (p < .05). The final testicular end velocity was lower in animals with long/moderate-shaped (L/M) (7.31 ± 2.91) than in those moderate/oval-shaped (M/O) (11.48 ± 3.88) testicles (p < .05). Animals with L/M testes presented higher pulsatility values and resistivity indices (0.51 ± 0.05; 0.55 ± 0.04) compared to animals with M/O shape (0.29 ± 0.20; 0.36 ± 0.15). We showed that the blood flow of the supra testicular artery between the two evaluated ages differed, and that 24-month-old bulls presented better thermoregulation capacity. Animals with a long/moderate testicular format presented a greater vascular resistance, which was imposed on the blood flow due to the anatomical differences in the testicular artery, resulting in lower velocity, and indicating better heat dissipation in this format.