The use of ultrasonography in the reproductive evaluation of boars

The objective was to study the use of ultrasound as a complementary test in the breeding soundness evaluation in male pigs and study the pattern of echogenicity of the testicular parenchyma in boars of different racial groups. Twenty‐six adult boars from four different racial groups were used, 10 from the Piau breed (group 1), four from the commercial and finishing group (group 2), six Pietrain breed (group 3) and six from the Duroc breed (group 4). All animals were evaluated for breeding soundness evaluation and the ultrasound examination of the testicles. The groups of animals that were evaluated showed no difference in the main semen parameters that were evaluated, except for the sperm volume, concentration of the ejaculated sperm and the supravital staining; the lowest figures were for the animals from the Piau breed (group 1). In relation to the testicular biometrics, Duroc animals (group 4) had a greater scrotal width compared to the other groups. But when we assessed the intensity of pixels of the testicles, there was a difference between groups. The groups 2 (finishing animals), 3 (Pietrain) and 4 had no difference between themselves. Group 3 had greater pixel intensity in relation to group 1. Of the 26 animals studied, five showed an abnormality during ultrasound evaluation, like hydrocele, hyperechoic mass in the testicular parenchyma, cyst in the head of the epididymis and the presence of fluid in the head and tail of the epididymis. The various animal groups studied did not differ in the principal reproductive parameters evaluated, showing that despite the great variability of reproductive traits between breeds and within the same breed, the breeding soundness evaluation, the more complete it is, is essential for the selection of breeders and the ultrasonography of the reproductive system becomes an important addition in this examination.     

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.     

Assessment of age-related morphological changes in the testes of post-hatch light ecotype Nigerian indigenous chicken

The age-related morphological changes of the testes in light ecotype Nigerian indigenous chicken were evaluated in this study using gross anatomical, histological and histomorphometric techniques. The results showed that the testes of 3- to 9-month-old birds were light pink while testes of sexually mature chicken were creamy white in colour. The left and right testicular weight, length, diameter, circumference and the organosomatic indices increased significantly (p < .05) with increasing age across the groups. Although the mean tubular diameter and epithelial height of the left and right seminiferous tubules increased significantly (p < .05) with age, the tubular diameter, epithelial height and luminal diameter did not vary significantly (p > .05) between the left and right testes of all the groups. The one-cell layer thick germinal epithelium of the left testes at 3 to 6 months old showed islands of cell proliferation that contained spermatogonia and spermatocytes. At 6 to 9 months, the left testes exhibited numerous early spermatids with occasional occurrence of late stage spermatids while the right testes showed scanty early stage spermatids. At 12 to 18 months, the germinal epithelia of both left and right testes were characterized by the presence of Sertoli cells, spermatogonia, primary spermatocytes, numerous early and late stage spermatids as well as spermatozoa. In conclusion, the morphological features highlighted in the present study show that at pre-pubertal periods, the left testes may develop faster than the right testes. However, both left and right testes may participate actively in the production of spermatozoa during the post-pubertal life.     

Assessment of the accuracy of testicular dysfunction detection in male donkey (Equus asinus) with the aid of colour-spectral Doppler in relation to plasma testosterone and serum nitric oxide levels

This study aimed to determine the usefulness of colour and pulsed Doppler modes for the accurate diagnosis of donkeys suffering from subfertility to determine whether testicular vascularity assessment could be an indicator for sperm functionality. The study sample was composed of 10 male donkeys with normospermia (control group) and 10 donkeys with hypospermia. Animals underwent scrotal circumference measurement, testicular Doppler examination, seminal evaluation, blood sampling and hormonal assay. Semen volume and concentration were significantly (p ≤ .05) lower in the subfertile group (30.25 ± 1.22 ml and 89.44 ± 2.55 × 10⁶/ml) as compared with the control group (82.76 ± 1.65 ml and 452.78 ± 1.25 × 10⁶/ml), and total sperm/ejaculation was significantly (p ≤ .05) higher in the normal donkeys (28.30 ± 2.32 × 10⁹/total ejaculated) as compared with the subfertile group. Intratesticular coloured area showed a marked decline in the hypospermic males. There was no significant difference between the two groups in testosterone level, although the normal group showed an increase in nitric oxide metabolites. Both Doppler indices of the three branches of the testicular artery were elevated significantly (p ≤ .05) in abnormal donkeys, whereas Doppler peak systolic and end-diastolic velocities were increased in the normal group. Male donkeys with subfertility demonstrated lower arterial vascularity parameters in the form of intratesticular coloured area and blood flow rate; therefore, the most optimal parameters for differentiating subfertile hypospermic from normospermic donkeys were found to be the two Doppler indices, velocities parameters, testicular blood flow rate and nitric oxide levels.     

睾丸生理功能的植物性神经调节研究进展

研究发现除了内分泌调控外,睾丸神经对睾丸的发育、雄激素的分泌以及精子的发生也发挥重要的调节作用,与睾丸的内分泌调节功能相比较人们对植物性神经在睾丸生理活动调节方面的认识还很少。论文从睾丸的植物性神经分布、精索神经对睾丸发育和睾丸生理功能的影响以及睾丸不同发育时期胆碱能受体和肾上腺素能受体的分布、神经递质NE和Ach对睾丸发育和功能的影响等方面做了详细综述,并对今后的研究方向和研究意义做了展望。     

Differentiation and development of Leydig cell, and induction of spermatogenesis during testicular differentiation in the Japanese eel, Anguilla japonica

The initial appearance and the development of Leydig cells (LCs), the sites of steroid hormone production in the testis, were investigated ultrastructurally during testicular differentiation in the Japanese eel, Anguilla japonica. In addition, the effects of a single injection of human chorionic gonadotropin (HCG; 5 IU g body weight^-1) on histological changes of the testes and serum 11-ketotestosterone (11-KT) were examined at various stages (15–18, 20–23, 26–29, 32–35, 38–41 and 46–50 cm body length (BL)) of testicular differentiation. Testicular differentiation was morphologically characterized by the development of loose connective tissue on the medial side in animals 18–29 cm in BL. Ultrastructurally, LCs were first identified in the loose connective tissue of the testis of the 23 cm fish. In the testes of fish over 32 cm, clusters of LCs were distributed throughout the interstitial region accompanying the increase in number of spermatogonia. In fish larger than 32 cm, spermatogenesis was induced by administration of HCG; serum 11-KT levels were also raised. On the other hand, there was no effect on spermatogenesis or serum 11-KT levels in fish less than 29 cm, or in the controls. These result suggests that morphological differentiation of LCs occurs in testis of the 23 cm eel, and subsequently, the testes of eels of BL more than 32 cm acquire the capability to produce steroid hormones.     

Isolation of pituitary glycoprotein gonadotropins from the grass carp (Ctenopharyngodon idell)

Gonadotropins (GTH) were isolated from pituitary glands of grass carp and their properties investigated. Acetone-dried pituitaries were extracted with ammonium acetate-ethanol and were then passed through cation exchanger, anion exchanger, and gel filtration column. Two glycoprotein gonadotropins (DE I and DE II) were isolated of MW 56,000 and 53,000 daltons, respectively, as determined by gel filtration technique. Isoelectric points of the two GTHs also differed from each other. Sialic acid contents were 3.64% and 1.47% for DE I and DE II, respectively. Both GTH's stimulated, in a similar manner, testicular androgen and ovarian estradiol synthesisin vitro.     

小鼠ES细胞在不同饲养层上培养nanog基因的 表达水平与生长行为

观察小鼠ES-D3细胞在MEF、新生牛睾丸支持细胞(nBTSCs)和新生兔睾丸支持细胞(nRTSCs)饲养层上生长4 d nanog基因的表达水平和生长行为。RT-PCR分析显示，MEF饲养层上培养4d的ES-D3 nanog基因含量高；nBTSC饲养层上培养nanog基因表达较低；而RTSC饲养层上培养nanog基因表达最低。结果显示：不同饲养层上的ES-D3细胞集落形态和培养4d分化程度有差异。在MEF饲养层上的ES-D3细胞培养4d，集落形态仍然很规则，细胞间紧密，集落表面少见大细胞，AKP染色强阳性。在nBTSC饲养层上的ES-D3细胞，集落界限清晰，AKP染色显示，集落大部分细胞呈阳性，少数的细胞集落为弱阳性，可见集落表面分散有较多的大细胞。在nRTSC饲养层上的饲养层上的ES-D3，集落隆起不明显，集落界限不清晰，大部分集落形态不太规则，AKP染色显示集落中部分细胞呈阳性，少数的细胞集落为全阴性，集落表面有较少的大细胞，集落周围有伸出的成纤维细胞。     

日本沼虾雄性生殖系统的研究─—雄性生殖系统的结构及发育

日本沼虾的雄性生殖系统由精巢、输精管、端壶腹和雄性生殖孔组成。精巢外被一层结统组织膜，内为许多生植小管，小管壁包括生精上皮和管壁上皮，生精上皮不断向管腔内增殖，形成不同发育阶段的雄性生殖细胞，输精管分为前输精管（细段）、中输精管（螺旋段）和后输精管（直段），其管壁均由高柱状上皮和矮柱状上皮构成，具有分泌功能；端壶胞壁肌肉层厚，其它结构与输精管壁相仿。精巢的发育季节性明显。     

Male pseudohermaphroditism in a Labrador Retriever,and a review of mammalian sexual differentiation

CASE HISTORY: An 8-month-old Labrador Retriever was referred with a history of ambiguous external genitalia.

CLINICAL FINDINGS AND TREATMENT: Clitoromegaly within apparent vulval folds, and an adjacent subcutaneous mass were noticed on external examination. An intra-abdominal testicle, with epididymis and suspected vas deferens ducts, was found during exploratory celiotomy. Incision over the subcutaneous mass revealed the accompanying testicle. Clitoridectomy was performed and an os clitoris removed. Normal juvenile testes were diagnosed on histology of the gonads. Chromosomal studies revealed a normal 78, XY male chromosomal constitution. Due to the combination of a male karyotype (78, XY), the presence of testicular tissue in the gonads, and the appearance of the external genitalia, a diagnosis of male pseudohermaphroditism (MPH) was made.

CLINICAL RELEVANCE: This case presents the first report of MPH in a Labrador Retriever, and highlights the diagnostic steps recommended when confronted with a dog with ambiguous external genitalia.