Changes in Testicular Interstitial Connective Tissue of Hamsters (Mesocricetus auratus) During Ageing and After Exposure to Short Photoperiod

The testicular interstitium of Syrian hamster (Mesocricetus auratus) was studied during ageing and in testicular regression after exposure to a short photoperiod, in relation to the interstitial cells and their connective tissue. This tissue was assessed histochemically using Masson's trichrome technique and the expression of Heat Shock Protein 47 (HSP‐47) and collagen IV (α5) was assessed in Leydig cells. Finally, an ultrastructural analysis of some cells of the testicular interstitium was made. Leydig cells were positive for HSP‐47 and collagen IV (α5). Ageing did not change the parameters studied while the short photoperiod altered the synthetic activity of Leydig cells. The positivity index of these cells for HSP‐47 was significantly higher in the regressed testis, but was lower for collagen IV (α5). During ageing no change were observed. Ultrastructural Leydig cells showed a discontinuous basal lamina that did not change during ageing. The basal lamina was not identified in Leydig cells regressed by exposure to a short photoperiod. In conclusion; the intertubular connective tissue suffers little change with age. By contrast, in the testis regressed after exposure to a short photoperiod the studied parameters related to the intertubular connective tissue were altered. These changes are probably related with the low synthetic activity of regressed Leydig cell.     

Efficiency of the Spermatogenesis in Zebu Bulls (bos taurus indicus)

The objective of the research was to evaluate the efficiency of the spermatogenesis through the morphology of the testicular parenchyma in bulls of different zebu breeds. We used testicular fragments from bull of the breeds Nelore (n = 10), Polled Nelore (n = 6), Gyr (n = 5), Guzerat (n = 5) and Tabapuã (n = 5). The tissue was perfused with Karnovsky solution, included in glycol methacrylate and stained with toluidine blue–sodium borate 1%. Animals of the Nelore breed presented higher population of primary spermatocyte in pre‐leptotene/leptotene (38.30) and in pachytene (38.14) and round spermatids (113.30), higher yield of spermatogonia mitosis (21.2) and higher daily spermatic production per gram of testicular parenchyma (32.8 × 10⁶) than those from breeds Gyr, Guzerat and Tabapuã and higher general yield of spermatogenesis (62.4) than breeds Gyr and Tabapuã. There was no significant difference in any of the evaluated parameters between breeds Nelore and Polled Nelore. The rate of Sertoli cells did not vary between the studied breeds. Apparently, the genetic selection applied to the breeds has been improving the yield in the spermatogenic process by decreasing cellular loss, although it did not increase the support capacity of the Sertoli cells.     

INSL-3 protein expression in normal and cryptorchid testes of Ziwuling black goats

Ziwuling black goats are typically found in loess plateaus regions and the Ziwuling Nature Reserve. Cryptorchidism is a common disease in this inbred goat, and its pathogenesis has been linked with the expression of insulin-like factor 3 (INSL-3). Therefore, this study aimed to investigate anatomical alterations caused by cryptorchism and the expression and distribution of INSL-3 in normal and cryptorchid testicular tissues. The testicular tissues of 6-month-old Ziwuling black goats were collected for microscopic analyses using histochemical, immunohistochemical, immunofluorescence and biometrical methods, as well as Western blotting to compare the expression and distribution of INSL-3. A lower expression of INSL-3 was observed in cryptorchid compared with normal testicular tissues (p < .01). Cryptorchidism caused a significant reduction in layers of spermatogenic epithelium and tubule areas in Ziwuling black goat (p < .01). The interstitial to seminiferous tubule area ratio was larger in cryptorchid than in normal group. Periodic Acid-Schiff (PAS) staining revealed pronounced positive bands in the interstitial tissue, while positive Alcian blue (AB) staining was not clear, and AB-PAS staining revealed a positive red band in the basement membrane of cryptorchid group. Immunofluorescence revealed a strong signal of INSL-3 expression in Sertoli and peritubular myoid cells, and moderate signal in Leydig and spermatogenic cells in the normal group. However, in cryptorchid testicular tissues, the signal of INSL-3 expression was strong in primary spermatocytes, occasional in Sertoli cells, limited in Leydig cells and absent in peritubular myoid cells. Furthermore, immunohistochemistry showed that INSL-3 expression was higher in normal testes compared with cryptorchid testicular tissues (p < .05), especially in primary spermatocytes and Sertoli cells. Collectively, our results indicate that cryptorchidism is closely related to the disorder of acid glycoprotein metabolism and the reduction in release of INSL-3 from Leydig cells. Moreover, Sertoli and peritubular myoid cells are crucial for INSL signalling and could underpin further research on the mechanism of cryptorchidism in animal.     

Asymmetrical size and functionality of the pampas deer (Ozotoceros bezoarticus) testes: Right testis is bigger but left testis is more efficient in spermatogenesis

Information about gonadal asymmetries in ruminants is very scarce. In this work, we performed three complementary studies to compare characteristics of both testes: (i) weight and size of offspring and adult dead males; (ii) the tissue:fluid relationship determined by ultrasound scanning; and (iii) the spermatogenic status using fine needle aspiration cytology. The right testis was heavier than the left one in both offspring and adult animals and had greater width and volume in adult males than the left one. The ultrasound pixel intensity was similar in both testes. The right testis tended to have more spermatogonia (p = .06) and had a greater percentage of early spermatids (p = .004) than the left testis. On the other hand, the left testis had a greater percentage of spermatozoa (p = .05). The left testis had a greater spermatozoa/spermatogonia ratio (p = .02) and tended to have more spermatozoa/Sertoli cells ratio (p = .07). The spermatogenic index tended to be greater in the left than in the right testis (p = .06). Overall, we concluded that the right testis of pampas deer males is bigger but according to the cytology, it seems to be less spermatogenically effective than the left one, but these differences are not explained by different tissue:fluid ratio in each testis. Although differences were greater in adults than in offspring, asymmetry was observed even in just born offspring.     

Seasonal changes in spermatogenesis and testicular steroidogenesis in wild male raccoon dogs (Nyctereutes procynoides)

Testes of 15 wild adult male raccoon dogs (Nyctereutes procynoides) obtained from September 2000 to April 2001 were studied to clarify seasonal changes in spermatogenesis and testicular steroidogenesis. There were marked seasonal variations in the testis weight and size with values relatively low in September and highest in March. Spermatogonia and primary spermatocytes were observed in September, while spermatogonia, spermatocytes and round spermatids were present in January, and all types of spermatogenic cells including mature spermatozoa were found in the mating season (February and March). The number of spermatogenic cells reached their peak values in February and March. In addition, steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3 betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17), and human placental aromatase cytochrome P450 (P450arom). P450scc and P450c17 were identified in Leydig cells and spermatids in February, whereas these enzymes were present only in Leydig cells in September. 3betaHSD was found in Leydig cells in September and February with more intense staining in February. The localization of P450arom changed seasonally: no immunostaining in September; more extensive immunostaining in Leydig cells, Sertoli cells, and elongating spermatids in February. These results suggest that seasonal changes in the testis weight and size of wild male raccoon dogs are correlated with changes in spermatogenesis. Seasonal changes in testicular steroidogenesis suggest that the synthesis of androgen and estrogen reaches its peak in the mating season.     

Morphometric Studies on the Testis of Korean Ring-necked Pheasant (<Emphasis Type="Italic">Phasianus colchicus karpowi</Emphasis>) during the Breeding and Non-breeding Seasons

The purpose of this study was to obtain detailed quantitative information on all cell types in the testis interstitium of Korean ring-necked pheasants and to combine these data with changes in the steroidogenic function of the testis during the breeding and non-breeding seasons. For animals collected during the breeding season, their testis weights, sperm production, serum testosterone levels and leuteinizing hormone (LH)-stimulated testosterone secretion were significantly (p < 0.01) increased compared to the non-breeding season. Testes of the pheasants during the non-breeding season displayed a 98% reduction in testis volume that was associated with a decrease in the absolute volume of seminiferous tubules (98% reduction), tubular lumen (100%), interstitium (90%), blood vessels (84%), lymphatic spaces (97%), Leydig cells (79%), mesenchymal cells (51%) and myoid cells (61%) compared to the breeding season. The numbers of Leydig cells, mesenchymal cells and myoid cells per testis in the breeding season were much higher than in the non-breeding season. Although the mean volume of a Leydig cell was 74% lower in the non-breeding season, the mean volumes of myoid and mesenchymal cells remained unchanged. These results demonstrate that there are striking differences in the testicular structure of the Korean ring-necked pheasant during the breeding and non-breeding seasons. Every structural parameter of the Leydig cell was positively correlated with both testosterone serum levels and LH-stimulated testosterone secretion. The correlation of changes in hormonal status with the morphometric alterations of Leydig cells suggests that the Korean-ring necked pheasant may be used as a model to study structure-function relationships in the avian testis.     

Comparison of the Distribution of PGP9.5 and NPY in Cryptorchidism and Testicular Tumors in Dogs

The purpose of this study was to analyze the distribution and expression of peptidergic neurotransmitters protein gene product 9.5 (PGP9.5) and neuropeptide Y (NPY) in cryptorchidism and testicular tumors of dogs,compare them with normal testicular tissues of the same age,and provide reference for clinical diagnosis of malignant transformation in testicular tumors of dogs.HE staing,Masson trichrome staining,Gomori silver staining and toluidine blue staining were used to observe the tissue characteristics of reticular fibers,collagen fibers and mast cells.Immunohistochemical SP method and immunofluorescence combined with IPP were used to analyze the expression and localization of PGP9.5 and NPY in tissues.The results showed that the seminiferous epithelium of normal dog testis was composed of 4-7 layers of spermatogenic cells and Sertoli cells,and the distribution of collagen fibers and reticular fibers in interstitial tissue was sparse.The thickness of collagen fibers in the basement membrane of cryptorchidism seminiferous tubules increased,the nucleus of Sertoli concentrated at the base of seminiferous tubules,and the interstitial reticular fibers increased.The tissue structure of testicular tumor was unclear,collagen fibers and reticular fibers were irregularly distributed,and mast cells increased significantly compared with normal and cryptorchid groups.The immunofluorescence results showed that PGP9.5 was moderately positive in Leydig cells of normal testis,no significant expression in spermatogenic cells,strongly positive in Leydig cells and spermatogenic cells of cryptorchidism,and occasional expression in testicular tumors.NPY was occasionally expressed in normal testicular Leydig cells,but not in spermatogenic cells,strong positive expression in Leydig cells and seminiferous epithelium of cryptorchidism,high density and strong positive expression in interstitial vessels,and no obvious expression in testicular tumors.Immunohistochemical statistics showed that the expression of PGP9.5 and NPY in testicular tumor tissue were extremely significantly lower than that in normal group (P<0.01),while the expression of PGP9.5 and NPY in cryptorchidism group were significantly or extremely significantly increased (P<0.05;P<0.01).Therefore,the expression of PGP9.5 and NPY in cryptorchidism of dogs was increased suggesting that the cryptorchidism of dogs had a tendency to develop into a tumor,and was related to the degree of malignant transformation of tumor.     

犬隐睾及睾丸肿瘤中PGP9.5和NPY的分布比较

试验旨在分析肽能神经递质蛋白基因产物9.5（protein gene product 9.5,PGP9.5）和神经肽Y（neuropeptide Y,NPY）在犬隐睾及睾丸肿瘤中的分布和表达,并与同年龄正常睾丸组织进行比较,为认识犬睾丸肿瘤恶变临床诊断提供参考。应用HE染色、Masson三色染色、Gomori银浸染、甲苯胺蓝染色观察各组织中网状纤维、胶原纤维及肥大细胞等组织特征,采用免疫组织化学SP法及免疫荧光法结合IPP统计分析PGP9.5和NPY在组织中的表达及定位。结果显示,正常犬睾丸生精上皮由4~7层生精细胞及Sertoli细胞构成,间质组织胶原纤维和网状纤维分布稀疏。隐睾生精小管基底膜胶原纤维厚度增加,Sertoli细胞核浓缩位于生精小管基底,间质网状纤维增多。睾丸肿瘤组织结构不清晰,胶原纤维和网状纤维无规则分布,肥大细胞较正常组及隐睾组显著增多。免疫荧光定位表明,PGP9.5在正常睾丸Leydig细胞中呈中等阳性表达,生精细胞中无明显表达;隐睾Leydig细胞及生精细胞中呈强阳性表达;睾丸肿瘤中偶有表达。NPY在正常睾丸Leydig细胞中偶见阳性表达,生精细胞中无表达;隐睾Leydig细胞及生精上皮中无表达,间质小血管管壁呈高密度强阳性表达;睾丸肿瘤组织中无明显表达。免疫组化统计表明,睾丸肿瘤组织中PGP9.5和NPY较正常组极显著降低（P<0.01）,隐睾组PGP9.5和NPY表达显著或极显著增加（P<0.05;P<0.01）。因此,犬隐睾时PGP9.5及NPY的表达增高,提示犬隐睾时已有发展为肿瘤的趋势,且与肿瘤恶变程度相关。     

老龄哈多利系博美犬睾丸组织结构分析

为比较哈多利系博美犬老龄与青年阶段睾丸的组织结构差异及相关蛋白分布特征，探索博美犬不同年龄阶段睾丸组织结构变化及其对生殖功能的影响，本试验应用特殊染色、免疫组织化学法结合免疫荧光观察比较青年与老龄博美犬睾丸组织化学特点，并用IPP图像分析软件进行定量分析。结果显示，与青年博美犬相比，老龄博美犬生精上皮层数与厚度降低，Leydig细胞数及Sertoli细胞数显著增加（P<0.05），生精小管基底膜及血管管壁层胶原纤维与网状纤维含量明显增加。免疫组织化学结果显示，老龄博美犬睾丸细胞外基质（exreacellular matrix，ECM）相关蛋白Ⅳ型胶原（collage，Col Ⅳ）、硫酸乙酰肝素蛋白多糖（heparan sulfate proteoglycan，HSPG）含量极显著低于青年博美犬（P<0.01），层黏连蛋白（laminin，LN）含量显著低于青年博美犬（P<0.05）。免疫荧光结果显示，Col Ⅳ、HSPG和LN主要在Leydig细胞及Sertoli细胞强表达。因此，老龄博美犬胶原纤维与网状纤维含量增加，Leydig细胞数及Sertoli细胞数增多可能与其生精功能的维持相关，其生精功能的下降与Col Ⅳ和HSPG的变化密切相关。     

Immunoexpression of Aromatase in Immature and Adult Males of the European Bison (Bison bonasus,Linnaeus 1758)

Based on recent literature dealing with the role of oestrogens in the male gonad, attempts were undertaken to reveal the site of aromatization within the testis of the European bison (Bison bonasus). Testes were collected from culled animals living in free‐ranging populations in Bialowieza Forest, Poland (nine males aged 8 months to 10 years). Moreover, to check for any alterations in the expression of testicular aromatase between American bison (Bison bison) and European bison, testes from one adult 10‐year‐old individual were also chosen for this study. For immunohistochemistry, 4% formaldehyde fixative was used. Both qualitative and quantitative evaluations of immunohistochemical staining were performed. Leydig cells, Sertoli cells and germ cells exhibited a positive immunoreaction for aromatase in testes of immature and sexually mature bison. A marked increase in aromatase expression was observed in three adult European individuals with impaired spermatogenesis. Consistent with recent data and those of our own, it might be suggested that the strong expression of aromatase negatively affects spermatogenic function in bison testes and may serve as a possible explanation of specific sperm defects observed in European bison bulls. On the contrary, one cannot exclude that differences in the aromatase immunoexpression levels are attributed to the homozygosity, the cause of frequent disease in European bison.     

Partial cDNA sequence of a relaxin‐like factor (RLF) receptor,LGR8 and possible existence of the RLF ligand‐receptor system in goat testes

Relaxin‐like factor (RLF), also known as insulin‐like factor 3 (INSL3), is produced by testicular Leydig cells, but its specific receptor LGR8 (leucine‐rich repeat family of G‐protein‐coupled receptor 8) has not been identified in goats. This study aimed to identify complementary DNA (cDNA) sequences of goat LGR8, and characterize the expression of both RLF and LGR8 in goat testes by RT‐PCR and immunohistochemistry. Testes were collected from immature (3‐month‐old) and mature (24‐month‐old) Saanen goats, and partial cDNA sequences of the goat homologue of human LGR8 were identified. The sequence encoded a reduced peptide sequence of 167 amino acids, which corresponded to transmembrane regions 2 through 5, followed by the beginning of intracellular loop 3 of human LGR8. Expression of both LGR8 and RLF genes was drastically increased in mature testes compared with immature ones. Although RLF protein was restricted to Leydig cells, LGR8 protein was detected in both Leydig cells and seminiferous epithelial cells (possibly germ cells and Sertoli cells). These results reveal a possible existence of the RLF‐LGR8 ligand‐receptor system within the goat testis, suggesting that RLF may play a role in testicular function through LGR8 on Leydig cells and seminiferous epithelial cells in an autocrine and/or paracrine manner.     

Abnormal functions of Leydig cells in crossbred cattle–yak showing infertility

In Mongolia, yak (Bos grunniens) are able to live in alpine areas and their products greatly influence the lives of the local people. Increased vigour in hybridized yak and cattle can offer benefits for livestock farmers. However, male hybrids show reproductive defects resulting from spermatogenesis arrest, affecting the conservation and maintenance of dominant traits in the next generation. The underlying mechanisms involved in hybrid cattle–yak infertility have recently been investigated; however, the genetic cause is still unclear. Androgens and androgen receptor (AR) signalling are required for spermatogenesis. We, therefore, evaluated the expression of AR, 3β-hydroxysteroid dehydrogenase (3βHSD) and 5α-reductase 2 (SRD5A2) in Leydig cells to investigate their function in cattle–yak spermatogenesis. Testicular tissues from yaks (1–3 years old) and hybrids (F1–F3, 2 years old) were collected and subjected to immunohistochemistry and image analyses to investigate the expression of each parameter in the Leydig cells. After maturation at 2 years, the expression levels of AR increased and the levels of 3βHSD decreased, but the SRD5A2 levels remained constant in yak. However, the cattle–yak hybrid F2 showed immature testicular development and significantly different expression levels of AR and 3βHSD compared with mature yak. These results suggest that the decreased expression of AR and increased expression of 3βHSD in the Leydig cells of cattle–yak hybrid testes may represent one of the causes of infertility. Our study might help in solving the problem of infertility in crossbreeding.     

Localization of Insulin‐Like Growth Factor‐I (IGF‐I) and IGF‐I Receptor (IGF‐IR) in Equine Testes

The insulin‐like growth factor‐I (IGF‐I) is a key regulator of reproductive functions. IGF‐I actions are primarily mediated by IGF‐IR. The main objective of this research was to evaluate the presence of IGF‐I and IGF‐I Receptor (IGF‐IR) in stallion testicular tissue. The hypotheses of this study were (i) IGF‐I and IGF‐IR are present in stallion testicular cells including Leydig, Sertoli, and developing germ cells, and (ii) the immunolabelling of IGF‐I and IGF‐IR varies with age. Testicular tissues from groups of 4 stallions in different developmental ages were used. Rabbit anti‐human polyclonal antibodies against IGF‐I and IGF‐IR were used as primary antibodies for immunohistochemistry and Western blot. At the pre‐pubertal and pubertal stages, IGF‐I immunolabelling was present in spermatogonia and Leydig cells. At post‐pubertal, adult and aged stages, immunolabelling of IGF‐I was observed in spermatogenic cells (spermatogonia, spermatocyte, spermatid, and spermatozoa) and Leydig cells. Immunolabelling of IGF‐IR was observed in spermatogonia and Leydig cells at the pre‐pubertal stage. The immunolabelling becomes stronger as the age of animals advance through the post‐pubertal stage. Strong immunolabelling of IGF‐IR was observed in spermatogonia and Leydig cells at post‐puberty, adult and aged stallions; and faint labelling was seen in spermatocytes at these ages. Immunolabelling of IGF‐I and IGF‐IR was not observed in Sertoli cells. In conclusion, IGF‐I is localized in equine spermatogenic and Leydig cells, and IGF‐IR is present in spermatogonia, spermatocytes and Leydig cells, suggesting that the IGF‐I may be involved in equine spermatogenesis and Leydig cell function as a paracrine/autocrine factor.     

The ultrastructure of spermatogenic cells and morphological evaluation of testicular development in the silver pomfret (Pampus argenteus)

The silver pomfret (Pampus argenteus) is a widely distributed and economically important marine fish in the Indo-Pacific. In this study, we acquired the second generation of wild P. argenteus by artificial breeding and further studied the testicular development and ultrastructure of spermatogenesis. The results of gonadosomatic index (GSI) showed the spawning period of this marine fish was from April to June. Besides, through morphological analysis, we found that P. argenteus had an anastomosing tubular testis surrounded by a layer of tunica albuginea, in which spermatogenesis occurred in cysts where the synchronous germ cells were completely surrounded by the cytoplasmic projection of Sertoli cells. Meanwhile, based on submicroscopic characteristics, the germ cells are classified into nine different types. During the ontogenesis of testis, both the early stage of spermatogenesis and sperm were observed in P. argenteus. At sperm maturation stage, different types of spermatozoa and activation of sperms occurred non-synchronously in the tubules. Cytoplasmic bridges also were observed among synchronous germ cells within the cysts, suggesting an interrelated and differentiated relationship among these germ cells.     

Morphology and Morphometry of Seminiferous Tubules in Crioulo Horses

This study aimed to assess the biometrics of the testes and the morphology of the seminiferous tubules of Crioulo horses. We studied 10 sexually mature stallions (3–6 years of age). After orchiectomy, testes were perfused with Karnovsky's solution and then embedded in glycol methacrylate. Testis sections (4 μm) were cut and stained with toluidine blue and a solution of 1% sodium borate. The histological images were digitized, and the morphometric analysis was performed using ImageJ software. The average weight of the stallions was 377.5 kg, and the average weight of both testicles was 162.9 g. The percentage of testicular parenchyma occupied by the seminiferous tubules and the intertubular tissue was 77.97% ± 6.34% and 22.03% ± 6.34%, respectively. The average tubular diameter was 205.00 ± 36.91 μm, whereas the average height of the seminiferous epithelium was 70.56 ± 2.82 μm. Average tubular length per testicle and average tubular length per gram of testicle were 4,085.10 ± 1,170.68 m and 26.09 ± 10.63 m/g, respectively. The characteristics of the eight stages of the seminiferous epithelium cycle were similar to those reported in other horse breeds. We conclude that the morphometry of the seminiferous tubules of Crioulo horse resembles what has been reported in other horse breeds. The volumetric proportion of the seminiferous tubules and the Leydig cells of the Crioulo horse is one of the highest ever reported for stallions.     

Testis Morphometry and Stages of the Seminiferous Epithelium Cycle in an Epididymal Sperm‐storing Neotropical Vespertilionid,Myotis levis (Chiroptera)

Yellowish myotis, Myotis levis, is a seasonal, epididymal sperm‐storing Neotropical vespertilionid. In the dry season, males show simultaneous testis regression and sperm storage in cauda epididymis, enabling them to mate during this season. In this study, we investigated seasonal variations in body mass, diameter and height of seminiferous tubules and nuclei of Leydig cells in a population of southeastern Brazil. We also determined the frequencies of the stages of the seminiferous epithelium cycle (SEC) of mature individuals of this population. Body mass and diameter of Leydig cell nuclei showed no significant differences between dry and rainy seasons and stages of annual reproductive cycle; however, all other morphometric parameters varied significantly. The relative cumulative frequency of pre‐meiotic stages of the SEC (1–3) was 51%, of meiotic stage (4) was 2% and of post‐meiotic stages (5–8) was 47%. We confirmed that the yellowish myotis presents seasonal sperm production as revealed by testis regression and epididymal sperm storage during the dry season.     

Relationship between angioarchitecture of the testicular artery and spermiogram parameters in Egyptian buffalo bulls (bubalus bubalis)

The current study aimed to investigate the effect of testicular artery angioarchitecture on the spermiogram parameters in Egyptian Buffalo bulls. Eight adult buffalo bulls aged between 2 and 8 years were used for semen evaluation. For anatomical studies, the masculine gonads were collected after slaughtering 30 adult bulls and prepared for injection by different masses (Urographine^®, Latex and Epoxy) through the testicular artery. The mass activity of the ejaculate was assessed immediately after collection. The sperm motility in fresh bull ejaculate was more than 80%. The overall mean percentage of sperm abnormalities was <18%. The recorded sperm abnormalities were mostly secondary one including distal protoplasmic droplet, fragmented tail, detached head, detached galea capitis and bent tail. The highest percentage of sperm viability was recorded just after sperm collection (alive > 85%). The results revealed that testicular artery can be divided into three parts (abdominal, funicular and marginal parts) along its course. The coils of the funicular part forming a cone-like structure with its base fixed to the head of the testis. Two epididymal branches to the head and tail of epididymis emanate from the funicular part which continues as pars marginalis on the lateral surface of testis before its division into the lateral and medial testicular arteries on approaching the tail extremity of the testis. The increase in the length of the testicular artery with increase in the size of the testes played a great role in the degree of complexity of the architectural vascular patterns. The degree of complexity is affected by the number of coils formed by the vessel. The increase in the convolutions of the vessel will reduce the speed of blood flow to the gonads. Thus in turn will enabling the thermoregulatory mechanism to work more efficiently and will affect the semen value.     

Examination of jackass (Equus asinus) accessory sex glands by B-mode ultrasound and of testicular artery blood flow by colour pulsed-wave Doppler ultrasound: Correlations with semen production

The accessory sex glands play a major role in the production of seminal plasma, and testicular artery blood flow seems to strongly influence testicular function. However, very little ultrasound imaging of these organs has been undertaken in donkeys. The present work reports the results of such examinations in five jackasses along the year. The accessory glands were inspected by B-mode ultrasound while the testicular artery blood flow was assessed by colour pulsed-wave Doppler ultrasound. The testicular artery was examined at pampiniform plexus (PPT), supratesticular area (ST) and capsular artery (CA). Values were recorded for the total arterial blood flow (TABF), peak systolic velocity (PSV), end-diastolic velocity (EDV), resistive index (RI), pulsatility index (PI) and time average maximum velocity (TAMV). Semen was obtained and assessed for sperm concentration, viability, abnormalities and motility using a CASA system. The bulbourethral glands, prostate and ductus deferens ampullae were relatively larger than in the stallion. Bulbourethral glands and ampullae sizes were inversely correlated with sperm motility. An reduction in blood flow between the level the PPP and the CA was observed, helping to reduce testis temperature and oxygen pressure. Blood flow at the CA showed the strongest correlation with semen production. PI and RI were positively correlated with the CASA motility variable STR (p = .02, p = .06) and sperm viability (p = .01), while sperm concentration (p = .05) correlated inversely with PSV, EDV, TAMV and TABF. EDV also correlated negatively with the CASA variables VSL, LIN, STR and VAP (p ≤ .05). PI and RI were also negatively correlated with testis length (p = .0093, p = −.0438).