Reference histology of veal calf genital and endocrine tissues - an update for screening on hormonal growth promoters

An animal experiment has been performed with 42 veal calves, 21 males and 21 females, which were fed and housed according to European regulations for veal calves. The animals were kept in six groups of seven animals and fed milk replacer supplemented with three different levels of lactulose (0%, 1% and 3%) and some roughage. At the start of the experiment the animals were 1-3 weeks of age and they were slaughtered at 26 weeks. From male animals prostate, bulbo-urethal gland and testes were sampled, from female animals Bartholin's gland, uterus, cervix and ovaries were sampled. From all animals thyroid, thymus, adrenals, liver and kidneys were sampled. Histological investigation of the prostates and bulbo-urethral glands showed normal histology. This means that dilated tubules, strong secretion, increased mucinous glandular tissue and severe hyperplasia and squamous metaplasia, as is regularly observed in practice in the Netherlands, were not present in these animals. None of these prostates would be judged as positive in the screening for hormones as is performed by the Dutch Food and Consumer Product Safety Authority (VWA). The female calves also showed normal histology of Bartholin's gland except for three animals that appeared to be in oestrus and showed some metaplasie of the ducts but with a normal gland to duct ratio. These animals would be judged as suspect. The liver and kidney showed minor alterations due to slight infections during the experimental period. This experiment showed that it is possible to raise veal calves according to the practice without getting positive histology in the prostate or Bartholin's gland.     

Effects of exposure to genistein and estradiol on reproductive development in immature male mice weaned from dams adapted to a soy-based commercial diet

Genistein, a soybean-originated isoflavone, is widely consumed by humans for putative beneficial health effects but its estrogenic activity may adversely affect the development of male reproductive system. Twenty one-day-old ICR mice weaned from dams fed with a soybean-based diet throughout gestation and lactation were exposed by gavage to genistein (2.5 mg/kg b.w./day) or 17beta-estradiol (7.5 microg/kg b.w./day) for five weeks. Corn oil was used as a negative control. The animals were fed with a casein-based AIN-76A diet throughout the experimental periods. There were no significant differences in body and organ weights of mice among experimental groups. No significant differences in sperm counts and sperm motile characteristics were found between control and genistein groups. Treatment of 17beta-estradiol caused a significant decrease in prostate weight and epididymal sperm counts compared to the control (p<0.05). The levels of phospholipid hydroxide glutathione peroxidase in the testis and prostate of mice exposed to genistein or 17beta-estradiol were significantly higher than that of the control mice (p<0.05). 17beta-estradiol treatment caused degeneration and apoptosis of germ cells in the testis, depletion and degeneration in the epididymal epithelium, and hyperplasia of mucosal fold region in the prostate of mice. Genistein treatment did not cause any lesion in the testis, epididymis, and prostate. These results suggest that dietary uptake of genistein during juvenile period may not affect male reproductive development and functions.     

Immunohistochemical study of galectin-3 in mature and immature bull testis and epididymis

Galectin-3, a member of the β-galactoside-binding protein family, has been implicated in mammalian sperm maturation. We examined galectin-3 expression in the testis and epididymis of sexually mature and immature bulls. Western blot analysis showed varying levels of galectin-3 in the bull testis and epididymis, and galectin-3 immunoreactivity was higher in the mature testis and epididymis than in immature organs. Galectin-3 was primarily localized in interstitial cells of the immature bull testis and in the peritubular myoid and interstitial cells of the mature testis. In the immature epididymis head, galectin-3 was primarily in the principal and basal cells of the epithelium. In the mature epididymis head, moderate levels of galectin-3 were detected in the sperm, while low levels were found in the stereocilia, epithelium and connective tissue. In the immature epididymis body, moderate protein levels were detected in the principal cells, while lower levels were found in the basal cells. The mature epididymis body showed moderate levels of galectin-3 immunostaining in the stereocilia and epithelium, but low levels in the connective tissue. In the immature epididymis tail, only low levels of galectin-3 staining were found in the epithelium, whereas the mature epididymis tail showed high levels of galectin-3 in the principal cells, moderate levels in the basal cells and low levels in connective tissue. These findings suggest that galectin-3 expression plays a role in the maturation and activation of sperm in bulls.     

Endocrine-paracrine cells of the male urogenital apparatus: a comparative histochemical and immunohistochemical study in some domestic ungulates

Specimens of testis, excurrent duct including the male accessory glands and urethra, were studied in boars, bulls, horses and donkeys, in order to localize endocrine/paracrine cells. Silver impregnation methods were used to test the argentaffinity and/or argyrophilia of cells. Immunoreactivities to chromogranin A, 5-hydroxytryptamine, somatostatin, [met]- and [leu]- enkephalins, gastrin-releasing peptide, calcitonin gene-related peptide, neuropeptide Y, substance P, vasoactive intestinal peptide, beta-endorphin antisera were tested by a streptavidin-biotin method. In the testis, epididymis, ductus deferens and vesicular gland no endocrine cells were found in any of the animals studied. Chromogranin-A, serotonin, somatostatin and enkephalins were present in endocrine/paracrine cells in the surface or glandular epithelia, whereas all other antisera gave negative results. In the prostatic complex and the urethral epithelium, the most consistent number of endocrine cells was serotonin-immunoreactive. Few cells were also argentaffin and a very limited number of them showed argyrophily and chromogranin-A immunoreactivity. Somatostatin-and enkephalin-immunoreactive cells were rare in the bull and boar, absent in stallions. This comparative study carried out on different species of domestic ungulates has shown deeply different immunophenotypes, even comparing species that are in a very close zoological relationship with one another, such as the horse and the donkey.     

Ambiguous genitalia in a fertile, unilaterally cryptorchid male miniature schnauzer dog

A 7-year-old male miniature schnauzer dog with unilateral cryptorchidism was presented for elective orchiectomy. Surgery to remove the cryptorchid testis revealed a fully formed uterus with horns attached to both testis and the body and cervix terminating at the prostate gland. The gross and microscopic diagnosis for the genital tract was persistent Müllerian duct syndrome with unilateral cryptorchidism. Additional associated lesions included cystic endometrial hyperplasia and a solitary, intratubular seminoma within the undescended testis. Persistent Müllerian duct syndrome is rare among domestic animals but is more common in miniature schnauzer dogs because of inheritance as an autosomal recessive trait.     

Clinical,Ultrasonographic and Pathological Findings in a Bull with Segmental Aplasia of the Mesonephric Duct

On assessment for use in an AI stud, a 12‐month‐old bull was found to produce low volume ejaculates with 41% of the sperm having morphological abnormalities. No left epididymal tail was palpable and the head of the epididymis on the left was twice the size compared with the right. Ultrasound examination showed the left testis to contain a large central area of decreased echogenicity, which could be followed proximally to a 15‐mm echolucent lesion at the site of the epididymal head. Postmortem examination revealed a 15‐mm diameter cyst in the region of the left epididymal head, and absence of the body and tail of the epididymis. The mediastinum testis of the left testis was dilated, corresponding to the area of decreased echogenicity observed on ultrasonography. No left seminal vesicle was present and the ampulla was significantly smaller than the same structure on the right. Histological examination revealed incomplete or absent spermatogenesis involving the majority of seminiferous tubules in the left testis, and a small proportion of those of the right testis. The cystic structure at the site of the left epididymal head was lined by irregular, sometimes attenuated, epithelium and contained sparse spermatozoa. This case demonstrates the adverse impact, which segmental aplasia of the mesonephric duct had on the testicular and epididymal function of a bull, and highlights the importance of careful clinical assessment in its diagnosis.     

Ultrastructure of the Bull Seminiferous Tubule Supporting Cells with Special Consideration of the Cell Junctions

This study represents an electronmicroscopic examination of seminiferous tubule epithelium of the bull, with special attention to the cell types and junctional complexes between cells which contribute to the blood-testis barrier. In general seminiferous epithelium of the bull resembles that of other animals with some alterations in supportive cell types and organization of junctional complexes between epithelial cells. These differences are illustrated and discussed in regard to their possible functional significance.     

增精宝对环磷酰胺致小鼠生精障碍保护作用的研究

为评价增精宝对雄性动物生精作用的影响,采用环磷酰胺致小鼠生精障碍为动物模型,考察不同剂量增精宝对环磷酰胺致小鼠生精障碍的保护作用。结果显示,高剂量组对小鼠的附睾、睾丸、储精囊和前列腺均有较显著的促进作用;低剂量组精子总密度和精子活率均差异显著(P<0.05);高剂量组精子活率差异显著(P<0.05);对小鼠睾丸各倍体生精细胞比例的影响,高剂量组小鼠睾丸生精小管较为饱满,管腔内存在大量精子,生精上皮较厚,可见各级生精细胞排列整齐,界膜明显,睾丸间质细胞排列整齐。结论表明,增精宝两种剂量均能够显著促进环磷酰胺导致小鼠生精障碍的保护作用,高剂量组对雄性小鼠的作用尤为显著。     

Die Frühentwicklung der Gonaden und die Ontogenese von Rete testis und Tubuli seminiferi recti beim Huhn (Gallus domesticus)1

The early development of the gonads, and the ontogenesis of the rete testis and tubuli seminiferi recti in the chicken (Gallus domesticus) The primordium of the gonads can be discerned in the 4-day chick embryo by the elevation of the celomic epithelium to form germinal epithelium, and by the arrival of the primordial germ cells. Already in the 4½ day chick embryo there appears on the left side of the body, as a result of the first proliferation of the germinal epithelium, a subepithelial mesenchymelike cell conglomeration, which has been erroneously labelled in the literature a “urogenital union”. In contrast to the opinion expressed in the literature such a union does not appear until the 10th day, when the mesenchymal cells change to forerunners of rete cells which after hatching differentiate into rete-epithelial cells. The rete testis consists of intra and extratesticular transverse cisterns and of an intercalated and partically subdivided longitudinal cistern. The tubuli seminiferi contori end on the intracapsular lingitudinal cisterns either directly, or indirectly by intercalated tubuli siminiferi recti or intratesticular transverse cisterns.     

Pathological and biochemical studies on experimental hypothyroidism in growing lambs

Secondary iodine deficiency was experimentally produced in growing male lambs by oral administration of 50 mg/kg bodyweight of Thiourea daily for 3.5 months. At the end of the experiment the animals became weak, emaciated, anaemic, significantly reduced in body weight with facial oedema and alopecia at thigh, legs and abdomen. The clinical analysis showed significant reduction in erythrocyte and leucocyte numbers and in levels of triiodothyronine and testosterone at the end of the experiment. The histopathological picture of the thyroid gland revealed hyperplasia of the follicle-lining epithelial cells which project into the lumen. The lumens of the follicles are devoid of colloid. The testes showed ill-developed small, empty seminiferous tubulcs. In the liver, the hepatocytes showed degeneration and vacuolation with proliferation of Kupffer cells, which contain haemosiderin pigment. The kidney showed glomerular lipidosis with accumulation of haemosiderin pigment in the cytoplasm of the renal tubules. Hyperkeratosis of the epidermis associated with excessive keratin formation within the hair follicles was detected. In conclusion, deficiency of iodine causes hypothyroidism which leads to retardation of growth, reduced wool production and interferes with sexual maturity of growing male lambs.     

Mammomonogamus laryngeus (Railliet, 1899) infection in buffaloes in Rio Grande do Sul, Brazil

Parasites of the genus Mammomonogamus affect the respiratory tract of domestic animals. The present study was carried out to determine the presence of Mammomonogamus laryngeus infection and to analyze its lesions in infected buffaloes in Rio Grande do Sul, Brazil, between April and November 1999. The infection rate was 30.5%. In 32 infected buffaloes, the worm pairs collected per animal did not exceed 20. The microscopic diagnosis showed intense polypoid to intramucosal proliferation at the entrance to the pharynx, to which the parasites had adhered, with foci of multifocal hydropic degeneration of the epithelium or individual degeneration of epithelial cells with mild intraepithelial inflammatory infiltrate. The submucosa revealed intense lymphocyte infiltrate extending into the salivary glands. The submucosa also showed formation of structures that resemble lymphoid follicles.     

Prenatal development of the horse ovary]

To answer the many open questions concerning the development of the horse's ovary, first the prenatal development was investigated. It resulted that follicles derive from the germinal epithelium and its cords, whereas the Leydig cells and the rete blastema originate from the mesonephros. In the second third of pregnancy the Leydig cells undergo an enormous proliferation, in the last third they degenerate. However this degeneration is not connected with the postnatal development of the ovulation groove.     

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.     

Branchial cleft cyst in a bull.

Branchial cleft cyst was diagnosed in a 6-month-old Angus bull with a large swelling in the distal ventral neck region. A definitive diagnosis could not be attained from results of the clinical examination, radiography, and ultrasonography. Diagnosis was made from histologic examination of the surgically removed mass. Branchial cleft cysts are remnants of the branchial apparatus and are considered rare in domestic animals. The differential diagnosis should include the thymic form of bovine viral leukosis, thymoma, abscess, goiter, and thyroid gland tumors as well as other rare cysts that can develop in the same location.     

Morphologic features of Sertoli cells in the intra-abdominal testes of cryptorchid dwarf goats

Light and electron microscopy revealed an age-related progression of alterations of Sertoli cells in the intra-abdominal and scrotal testes of unilaterally cryptorchid West African dwarf goats between the ages of 1 and 30 months. Alterations in the scrotal testis were, however, maturational and included differentiation of Sertoli-to-Sertoli cell junctional specializations, profusion of smooth endoplasmic reticulum, convolution of nuclear profiles, development of vacuolar components of the nucleolus, and an overall change in cell shape in response to proliferation of germinal cells. Corresponding features were observed in Sertoli cells of the contralateral intra-abdominal testis, but the cytoplasmic features were transient because the cells degenerated progressively. Early changes included segregation of the smooth endoplasmic reticulum into compact masses composed of dense, narrow cisternae, dilatation of the rough endoplasmic reticulum cisternae into large, irregular profiles, atrophy of the Golgi complex, and accumulation of lipid droplets and lipofuscin granules. Many of these organelles and inclusions no longer were obvious in Sertoli cells of 12- to 15-month-old goats; rather, intracellular vacuoles and dilated intercellular spaces had become common. In the 24- to 30-month-old goats, Sertoli cells in the intra-abdominal testis contained mostly microfilaments and basally located mitochondria with circular cristae in dense matrices. The Sertoli-to-Sertoli cell junctional specializations were structurally intact. These results indicated that, in spite of the unfavorable intra-abdominal environment, Sertoli cells of the intra-abdominal testis, before their degeneration, had developed features similar to those of the scrotal testis.     

Expression of GPR54 Gene in Different Tissues and Cellular Localization in Testis of Sheep

The objectives of this study were to evaluate the expression of G-protein coupled receptor 54 (GPR54) in various tissues and the cellular localization of testis of 4 to 6-month-old sheep.In this experiment,mRNA expression was detected by Real-time fluorescence quantitative PCR.Cellular localization of GPR54 in testis was examined by immunohistochemistry.The results showed that GPR54 mRNA was expressed in all the tissues,and abundantly expressed in hypothalamus,pituitary gland and testis.The expression level gradually increased with the individual growth and development,the mRNA and protein expression of GPR54 in testis of 6-month-old was significantly higher than 4-month-old and 5-month-old (P<0.05);It was detected that GPR54 expression only in spermatogonia and a very number of primary spermatocytes in the testis by immunohistochemical staining,and it could detect significantly that the GPR54 expression in many split early sperm cells.The results demonstrated that it had close relationship between GPR54 and sexually mature of animals and the process of spermatogenesis in male animals.     

GPR54基因在绵羊不同组织中的表达及其在睾丸中的定位研究

试验旨在研究GPR54(G-protein coupled receptor 54)基因在绵羊不同组织中的表达及其在4~6月龄公羔睾丸组织中的表达特性及定位。本研究采用实时荧光定量PCR法检测各组织GPR54 mRNA表达规律,以及其在不同月龄公羔睾丸中的表达量,运用免疫组化技术对睾丸中GPR54基因进行定位分析。结果显示,GPR54在不同组织中均有不同程度的表达,其中在下丘脑、垂体和睾丸中大量表达;GPR54在4~6月龄绵羊公羔睾丸组织中呈阶段性表达,6月龄表达量显著高于4和5月龄(P<0.05);4月龄时,在精原细胞和极少数的初级精母细胞中检测到较弱的阳性信号;6月龄性成熟时,在睾丸中分裂早期的精子细胞检测到强阳性信号。综上表明,GPR54基因在绵羊不同组织中广泛表达,在下丘脑-垂体-睾丸生殖轴中发挥重要的作用,并与动物的性成熟及雄性动物精子发生过程有密切的关系。     

Pathophysiology of small testes in beef bulls: relationship between scrotal circumference, histopathologic features of testes and epididymides, seminal characteristics, and endocrine profiles

Tissue sections from testes and epididymides obtained from 17 young beef bulls with scrotal circumference (SC) between 27 and 40.5 cm were studied to determine whether small testes were a manifestation of lesions or a result of less, but otherwise normal, seminiferous epithelium. The SC correlated negatively with the estimates of germinal epithelial loss and positively with seminiferous epithelial area. Four bulls with SC less than 30 cm had severe lesions in their testes. Hypoplastic tubules were characterized by Sertoli's cells only with no evidence of germinal cells. Loss of germinal cells, leaving vacuolated epithelium and atrophy, were observed in degenerated tubules. Hyperplasia of Leydig's cells was observed in the vicinity of Sertoli's cell-only tubules, resulting either from degeneration or hypoplasia, and atrophy of Leydig's cells was associated with tubules devoid of Sertoli's cells. These findings indicated that Sertoli's cells may produce a factor(s) required for maintenance and regulation of Leydig's cell function. Epididymal epithelium, especially in the head, had regressed in bulls with hypoplastic and degenerative changes in their testes. Decreased sperm concentration and motility and an increased frequency of morphologic defects were observed in the 4 bulls with testicular lesions and regressed epididymal epithelium. Blood plasma profiles of cortisol, follicle-stimulating hormone, luteinizing hormone, and testosterone were determined in the 4 bulls with SC less than 30 cm and 10 of the 13 bulls with SC greater than 30 cm. There were no statistically significant (P greater than 0.1) differences in the responses to exogenous gonadotropin-releasing hormone or base-line patterns of blood plasma follicle-stimulating hormone and luteinizing hormone between the 2 groups. However, in the bulls with SC less than 30 cm, the mean concentration of testosterone was lower, whether spontaneous (P less than 0.05) or exogenous gonadotropin-releasing hormone induced (P less than 0.1). The fact that these bulls were not deficient in gonadotropins indicated that Leydig's cell function was impaired by local factors, either the factors that caused the tubular damage or those consequent to the tubular damage.     

Potential of triptolide in reproductive management of the house rat,Rattus rattus

Mature and healthy male house rats, Rattus rattus (n= 160) were fed on bait (cracked wheat: powdered sugar, 98:2) containing different concentrations of triptolide (0.1, 0.05, 0.025 and 0%) for 7 and 14 days in no‐choice and bi‐choice feeding tests in the laboratory. The objective of the study was to record the antifertility affects of triptolide after 30 and 60 days of termination of treatment. Results revealed no significant effect of triptolide treatment on weights of testis, epididymis, seminal vesicles and prostate gland of rats. Overall, sperm motility, live sperm count, sperm density and sperm morphology in the cauda epididymal fluid were found to differ significantly (P≤ 0.05) between untreated and treated groups of rats. The major effect of triptolide on sperm morphology was in the form of sperm head tail separation, which was up to 56.0% in rats treated for 14 days in no‐choice and autopsied after 30 days. A significant effect (P≤ 0.05) of triptolide treatment was observed on the histomorphology of the testis, which included a dose‐dependent decrease in diameter of seminiferous tubules, thickness of germinal epithelium and numbers of various spermatogenic cells. Cell associations in the seminiferous epithelial cycle were poorly developed in rats ingesting medium (4.7–5.1 mg/100 g bw) and high doses (6.9–7.2 mg/100 g bw) of triptolide than rats ingesting low doses (1.8–2.3 mg/100 g bw) and untreated rats. The cell stages affected had not recovered fully within the 60 day period following triptolide withdrawal. The present study suggests the potential of triptolide in reproductive management of Rattus rattus.