Morphological aspects and germ cells of male reproductive tract of river stingray, Potamotrygon amandae

The family Potamotrygonidae are the only species of stingrays restricted to fresh water and located exclusively in South America. The objective of this research was to analyze the morphological aspects and germ cells of the male reproductive tract of Potamotrygon amandae. The samples were fixed in 10% formalin, and then dehydrated in an ascending ethanol series (70 to 100%). To carry out light microscopy analyses, they were embedded in paraffin, cut and stained; as for scanning electron microscopy analyses, the samples were dried, glued in metallic bases and metalized. The gross morphology consisted of the following paired organs: testis, epididymis, deferent duct, Leydig gland, seminal vesicle, clasper, and the clasper gland. Microscopically, several stages of spermatogenesis were observed in the testis, occurring in spherical follicles, similar to other stingrays. The epididymis was formed by one duct subdivided in various tubules. The deferent ducts were continuous with the epididymis, and the lumen was full of spermatozoa. The Leydig glands consisted of glandular units with eosinophilic content in the lumen of some, and the deferent ducts ran parallel to the ventral portion. The seminal vesicles possessed numerous compartments to store the sperm, with a wall similar to a hive, and the lumen was full of spermatozoa. Alcian Blue (AB) and Periodic Schiff-Acid (PAS) performed in the Leydig Gland, deferens ducts and seminal vesicle was positive only in the connective tissue, the cilia were PAS+ and the nuclei stained weakly for AB. The clasper gland was composed of unit glands and was covered with striated muscle externally. It stained very well with Periodic Schiff-Acid. The morphological aspects of the male reproductive tract of Potamotrygon amandae were similar to other stingrays.(AU)

Bilateral testicular tumors in a non-cryptorchid dog: coexistence of Sertoli cell and interstitial endocrine cell tumors

Background: Testicular neoplasms in dogs are more frequent than in other animal species, representing the most common tumors in elderly subjects after skin neoplasms. In cryptorchid subjects, the risk of neoplastic degeneration is high. The cytological examination is essential to differentiate the type of neoplasia and to determinate the best diagnostic approach. Aim of this report was to describe clinical and histopathological features of a dog with coexistence of Sertoli cell and interstitial endocrine cell tumors in a non-cryptorchid dog. Case: A 9-year-old non-neutered male dog, German Spitz breed, was presented to the veterinary clinic. On clinical examination, the dog had a body condition (BCS 6/9), pink mucous membranes, capillary refill time (CRT) < 2 sec, lymph nodes of normal size, afebrile, normal heart and respiratory rate. The abdomen was depressible to palpation, without pain, the skin appeared hyperpigmented, with generalized presence of comedones, pendular foreskin and absence of hair at the abdominal level, in the ventral portion of the trunk and neck, scant hair also at the level of the inner thighs and in perianal. At ultrasounds examinations, the right testicle presented a hypoechoic circular focal lesion, in the caudal pole, of 0.7 cm in diameter, well defined, echogenicity and a remnant of normal echostructure, smooth and regular margins; the left testicle showed an increase in size, irregular margins, with a heterogeneous echo structure, given by hypoechoic areas, referred to cysts, hemorrhagic or necrotic areas. The hemogram reported slightly microcytic and normochromic regenerative anemia. The leukogram showed monocytosis. The absence of the typical stress leukogram characterized by neutrophilia, lymphopenia and eosinopenia, and the reduction of ALP allows to rule out Cushing's disease. In order to rule out hypothyroidism due to the inhibitory effect of estrogens on the release of thyroid-stimulating hormone (TSH) the concentration of total thyroxine was analyzed, reporting normal values excluding hypothyroidism. Blood oestradiol 17-ß (E2) concentration was increased, with a normal testosterone (T) concentration of 0.30 ng/mL. Given the suspicion of the presence of testicular tumors, castration was performed by the surgical excision of both gonads, after ruling out the presence of abdominal or pulmonary metastases by chest and abdominal radiography. Discussion: The clinical, histopathological findings supported the diagnosis of testicular tumors. According to the pathological report, both gonads presented parenchymal nodular neoplastic nodular areas referring to the Sertoli sustentacular cells in the right testicle, to the proliferation of Leydig interstitial cells in the left one. The nodule in the left testicle was unencapsulated and showed a solid-diffuse pattern. Neoplastic cells were irregular polygonal, medium to large in size with moderate nucleus/cytoplasm ratio and moderate anisocytosis. In the right testicle, the nodule was heterogeneous in consistency and a diffuse pattern was present. Neoplastic cells were polygonal morphology, had a moderate nucleus/cytoplasm ratio and were organized tubules lined and obliterated the extensive cell growth. Bilateral orchiectomy allowed to improve the clinical signs, and 3 months after surgery, the animal was in good health, with evident improvement skin lesions. The E2 analysis was repeated, detecting normal values, demonstrating that testicular neoplasm in this patient were involved in E2 production; also T concentration decreased considerably from 0.30 to < 0.07 values.

Intertubular morphometric and ultrastructural testes analyses in mdx mice

Duchenne Muscular Dystrophy (DMD) reproductive alterations and the influence of antioxidant treatments may aid in understanding morphometry testicular quantification. In this context, the aim of the present study was to characterize the intertubular compartment (ITC) morphometry of animal testes in mdx mice supplemented with ascorbic acid (AA). Sixteen mice were used, namely the C57BL/10 (non-dystrophic) and C57BL/10Mdx (dystrophic) lineages, distributed into the following groups: Control (C60), Dystrophic (D60), Control supplemented with AA (CS60), Dystrophic supplemented with AA (DS60). A total of 200 mg/kg of AA were administered to mice for 30 days. Subsequently, the testicles were collected, weighed, and fragmented. The obtained fragments were fixed in Karnovsky's solution (pH 7.2) and embedded in historesin for morphometric and transmission electron microscopy assessments. Leydig cells were hypertrophic in the D60 group, but was reverted by AA supplementation in the DS60 group. The DS60 group also exhibited increased intertubular volume compared to the CS60 group. The ultrastructural images identified multilamellar bodies in dystrophic animals (lipid storage) and telocyte cells (transport substances) in both control and dystrophic animals. Morphometric alterations were, therefore, noted in the intertubular compartment due to Duchenne muscular dystrophy (DMD), with AA administration capable of altering Leydig cells in this condition.(AU)

Expression of the NSE,SP,NFH and DβH in normal and cryptorchid testes of Bactrian camel

Abstract Neuroendocrine substances play essential roles in regulating the normal physiological functions of testicles. The purpose of this study is to explore the localization and effects of four neuroendocrine markers (NSE, SP, NFH and DβH) in normal and cryptorchid testes of Bactrian camels using western blotting, transmission electron microscopy, immunohistochemistry, and immunofluorescence methods. The results showed that cryptorchidism caused a reduction in layers of spermatogenic epithelium and decreased glycogen positivity in the basement membrane. The ultrastructure revealed that macrophages were always found around the Leydig cells, crowded with swelling mitochondria in cryptorchidism. Expression of NSE in the Leydig cells of cryptorchidism was significantly weakened compared to that in the normal group(p<0.01). We found that SP was always distributed along the nerve fibers in normal testes and was expressed in the Leydig cells of cryptorchidism. However, expression of NFH in the cryptorchidic tissue was strongly positive in the spermatogenic epithelium, with limited expression in Leydig cells and no expression in peritubular myoid cells. Therefore, the expression of DβH in the Sertoli cells was comparatively strong in both the normal and cryptorchidism groups. NFH and DβH expression was significantly increased in the cryptorchidism group compared with the normal group (p<0.01). These findings indicated that the underdeveloped seminiferous epithelium and pathological changes in cryptorchid tissue in Bactrian camels were potentially related to a disorder in glycoprotein metabolism. Our results suggest that NSE and SP could help judge the pathological changes of cryptorchidism. The present study provides the first evidence at the protein level for the existence of NFH and DβH in Sertoli and Leydig cells in Bactrian camel cryptorchidism and provides a more in-depth understanding of neuroendocrine regulation is crucial for animal cryptorchidism.

Expression of the NSE, SP, NFH and DbetaH in normal and cryptorchid testes of Bactrian camel

Neuroendocrine substances play essential roles in regulating the normal physiological functions of testicles. The purpose of this study is to explore the localization and effects of four neuroendocrine markers (NSE, SP, NFH and DβH) in normal and cryptorchid testes of Bactrian camels using western blotting, transmission electron microscopy, immunohistochemistry, and immunofluorescence methods. The results showed that cryptorchidism caused a reduction in layers of spermatogenic epithelium and decreased glycogen positivity in the basement membrane. The ultrastructure revealed that macrophages were always found around the Leydig cells, crowded with swelling mitochondria in cryptorchidism. Expression of NSE in the Leydig cells of cryptorchidism was significantly weakened compared to that in the normal group(p<0.01). We found that SP was always distributed along the nerve fibers in normal testes and was expressed in the Leydig cells of cryptorchidism. However, expression of NFH in the cryptorchidic tissue was strongly positive in the spermatogenic epithelium, with limited expression in Leydig cells and no expression in peritubular myoid cells. Therefore, the expression of DβH in the Sertoli cells was comparatively strong in both the normal and cryptorchidism groups. NFH and DβH expression was significantly increased in the cryptorchidism group compared with the normal group (p<0.01). These findings indicated that the underdeveloped seminiferous epithelium and pathological changes in cryptorchid tissue in Bactrian camels were potentially related to a disorder in glycoprotein metabolism. Our results suggest that NSE and SP could help judge the pathological changes of cryptorchidism. The present study provides the first evidence at the protein level for the existence of NFH and DβH in Sertoli and Leydig cells in Bactrian camel cryptorchidism and provides a more in-depth understanding of neuroendocrine regulation is crucial for animal cryptorchidism.(AU)

Glycerin supplementation strategies for three or seven days affects oxidative stress, follicle dynamics and ovulatory response in Morada Nova sheep

The Ziwuling black goat is an indigenously in China, their offspring are frequently affected by congenital cryptorchidism. The extracellular matrix (ECM) contains cytokines and growth factors that regulate the development of the testis, and component changes often result in pathological changes. Cryptorchidism is closely related to structural changes in ECM. In this study, the histochemical staining, immunohistochemical, immunofluorescence and Western blot combined with semi-quantitative analysis was used to describe the distribution of the important ECM components Collagen type IV (Col IV), laminin (LN)and heparan sulfate proteoglycans (HSPG) in the normal and cryptorchid testes of Ziwuling black goats. Results showed that: The histochemical staining showed that the dysplasia of seminiferous tubules and decreased number of Sertoli cells in cryptorchidism, as well as sparse collagen fiber. Meanwhile, the distribution of reticular fibers is relatively rich. Furthermore, the PAS and AB staining in the interstitial vessels and lamina propria of seminiferous tubules is weak. The immunohistochemical and immunofluorescence revealed that Col IV, LN was strongly expressed in Leydig, Sertoli cells of normal testes and moderately positive in the spermatogonia and spermatids, but HSPG was not expressed in the spermatogonia. However, cryptorchidism, the expression of Col IV, LN and HPSG in Leydig, Sertoli cells significantly decreased, as well as the expression of Col IV and LN in capillary endothelial cells, but HSPG was moderately expressed in spermatogonia. Based on these data, the underdevelopment of spermatogenic epithelium, decreased synthesis function of collagen fibers and Leydig cells develop usually in the cryptorchidism were shown to be closely related to the abnormal metabolism of Col IV and LN. The positive expressed of HSPG in the spermatogonia of cryptorchid testes is related to the compensatory development of spermatogonia.(AU)

Distribution of extracellular matrix related proteins in normal and cryptorchid ziwuling black goat testes

The Ziwuling black goat is an indigenously in China, their offspring are frequently affected by congenital cryptorchidism. The extracellular matrix (ECM) contains cytokines and growth factors that regulate the development of the testis, and component changes often result in pathological changes. Cryptorchidism is closely related to structural changes in ECM. In this study, the histochemical staining, immunohistochemical, immunofluorescence and Western blot combined with semi-quantitative analysis was used to describe the distribution of the important ECM components Collagen type IV (Col IV), laminin (LN)and heparan sulfate proteoglycans (HSPG) in the normal and cryptorchid testes of Ziwuling black goats. Results showed that: The histochemical staining showed that the dysplasia of seminiferous tubules and decreased number of Sertoli cells in cryptorchidism, as well as sparse collagen fiber. Meanwhile, the distribution of reticular fibers is relatively rich. Furthermore, the PAS and AB staining in the interstitial vessels and lamina propria of seminiferous tubules is weak. The immunohistochemical and immunofluorescence revealed that Col IV, LN was strongly expressed in Leydig, Sertoli cells of normal testes and moderately positive in the spermatogonia and spermatids, but HSPG was not expressed in the spermatogonia. However, cryptorchidism, the expression of Col IV, LN and HPSG in Leydig, Sertoli cells significantly decreased, as well as the expression of Col IV and LN in capillary endothelial cells, but HSPG was moderately expressed in spermatogonia. Based on these data, the underdevelopment of spermatogenic epithelium, decreased synthesis function of collagen fibers and Leydig cells develop usually in the cryptorchidism were shown to be closely related to the abnormal metabolism of Col IV and LN. The positive expressed of HSPG in the spermatogonia of cryptorchid testes is related to the compensatory development of spermatogonia.(AU)

Efeitos da castração imunológica no epitélio seminífero de machos bubalinos / Effects of immunological castration on the seminipheral epithelium of buffaloes males

O procedimento de imunocastração vem sendo utilizado como técnica alternativa amplamente favorável ao bem-estar animal, por ser indolor, pouco invasiva e com eficácia semelhante à da castração cirúrgica, que causa maior estresse aos animais, principalmente quando realizada de forma inadequada. A imunocastração estimula a produção de anticorpos contra o Hormônio Liberador de Gonadotrofina (GnRH), bloqueando temporariamente a produção da testosterona pelas gônadas masculinas. Face a escassez de informações sobre este procedimento em touros bubalinos, o estudo teve como objetivo analisar os possíveis efeitos da imunocastração no parênquima testicular de búfalos. Vinte touros bubalinos, com idade entre 2 a 3 anos, oriundos da ilha do Marajó ­ PA, foram avaliados e divididos aleatoriamente em dois grupos composto por dez animais no grupo controle (GC) e dez do grupo imunocastrado (GIM). O produto utilizado foi a vacina anti-GnRH Bopriva ® (Zoetis, SP, Brasil). O GIM recebeu duas doses de 1,0ml contendo 400µg da vacina Bopriva ® com intervalo de 8 semanas entre as aplicações e o grupo controle recebeu 1,0 ml de solução fisiológica. Após 14 dias da última dose, os animais foram abatidos e coletado os 20 pares de testículos para analises de parâmetros macroscópicos como: comprimento, largura, circunferência e peso. Além disso, foi realizada a retirada de fragmentos do parênquima testicular para confecção de lâminas histológicas. Para comparação dos dados foi aplicado o teste t de Student (Nonparametric Test) considerando a significância com p<0,05 e intervalo de confiança de 95%. Foi observado diferença significativa entre os dois grupos para grupos para peso (direto p<0001; esquerdo p≤ 0,0001), circunferência (direita p< <0,0001), 1; esquerda p<0,0001), ), largura (somente lado direito p<0,0372) e comprimento (direita p≤ 0,0013; esquerda p<0,0437). Na avaliação microscópica, os animais do GC não apresenteram alterações. Nos animais do GIM houve degeneração em todas as amostras, sendo visualizado descamação, tortuosidade e espessamento da membrana basal do túbulo seminífero, assim como vacuolização e atrofia das células de Sertoli. Foi observado também uma redução do número de células de Leydig, fibrose intertubular pronunciada, núcleos picnóticos, azoospermia e células multinucleadas no interior dos túbulos. Conclui-se a vacina anti-GnRH mostrou-se eficaz em provocar lesões no parênquima testicular, comprometendo significativamente a espermatogênese a partir da possível supressão de testosterona.(AU)

The immunocastration procedure has been used as an alternative technique widely favorable to animal welfare, as it is painless, minimally invasive and with similar efficacy to surgical castration, which causes greater stress to animals, especially when performed improperly. Immunocastration stimulates the production of antibodies against Gonadotropin Releasing Hormone (GnRH), temporarily blocking the production of testosterone by the male gonads. Due to the scarcity of information about this procedure in buffalo bulls, the study aimed to analyze the possible effects of immunocastration on the testicular parenchyma of buffaloes. Twenty buffalo bulls, aged between 2 and 3 years, from the island of Marajó - PA, were evaluated and randomly divided into two groups consisting of ten animals in the control group (CG) and ten from the immunocastrated group (GIM). The product used was the anti-GnRH vaccine Bopriva ® (Zoetis, SP, Brazil). The GIM received two doses of 1.0ml containing 400µg of Bopriva ® vaccine with an interval of 8 weeks between applications and the control group received 1.0 ml of saline solution. After 14 days of the last dose, the animals were slaughtered and the 20 pairs of testes were collected for analysis of macroscopic parameters such as: length, width, circumference and weight. In addition, fragments of the testicular parenchyma were removed for the preparation of histological slides. To compare the data, Student's t test (Nonparametric Test) was applied, considering the significance with p< 0.0372) and length (right p≤ 0.0013; left p<0.0437). In the microscopic evaluation, the animals of the CG showed no alterations. In the GIM animals there was degeneration in all samples, with desquamation, tortuosity and thickening of the seminiferous tubule basement membrane, as well as vacuolization and atrophy of Sertoli cells. A reduction in the number of Leydig cells, pronounced intertubular fibrosis, pyknotic nuclei, azoospermia and multinucleated cells within the tubules were also observed. In conclusion, the anti-GnRH vaccine proved to be effective in causing lesions in the testicular parenchyma, significantly compromising spermatogenesis from the possible suppression of testosterone.(AU)

Diagnóstico e tratamento videoassistido de tumor de células de Leydig em cadela com síndrome do ovário remanescente / Videoassisted diagnosis and treatment for Leydig cell tumor in a bitch with remaining ovary syndrome

Neoplasias ovarianas de origem primária são incomuns em cadelas, exigindo métodos diagnósticos e terapêuticos mais precisos. Fêmeas também possuem células intersticiais secretoras de esteróides, contudo o aparecimento do tumor de células de Leydig é considerado raro. Uma cadela sem raça definida e 12 anos apresentava sinais de estro após ovariohisterectomia pré-púbere. Na avaliação ultrassonográfica, foi identificada uma massa localizada na região abdominal lateral direita, sugestiva de ovário remanescente. Durante a inspeção laparoscópica foi identificada a massa distante caudalmente do coto ovariano direito, inserida em região de mesentério jejunal. O segmento intestinal foi apreendido e exteriorizado de maneira videoassistida, optando-se pela remoção do tumor associado à enterectomia do segmento afetado. A avaliação do coto ovariano esquerdo, revelou um cisto de 0,5cm de diâmetro que também foi removido. A análise histopatológica foi sugestiva de neoplasia de células intersticiais (tumor de células de Leydig) e um cisto epitelial ovariano. O tumor de células de Leydig é relatado com freqüência em machos, entretanto, sua aparição em fêmeas é considerada extremamente incomum acometendo principalmente animais não esterilizados. A neoplasia não aderida ao coto ovariano comprova ainda que resquícios de tecido ovariano que porventura fiquem livres na cavidade abdominal podem ser responsáveis pelo desenvolvimento da síndrome do ovário remanescente e neoplasias. O prognóstico para neoplasias ovarianas sem metástases é favorável, principalmente quando aplicada a intervenção cirúrgica como método terapêutico. O uso da videocirurgia para auxiliar no diagnóstico e tratamento demonstrou-se adequado para o caso descrito, mas que deve ser complementado pela análise histopatológica.

Primary ovarian neoplasms are uncommon in bitches, requiring more accurate diagnostic and therapeutic methods. Females also have steroid-secreting interstitial cells, however, the appearance of the Leydig cell tumor is considered rare. A 12-year-old mongrel bitch showed signs of estrus after prepubertal ovariohysterectomy. Ultrasound evaluation located a mass in the right lateral abdominal region, suggestive of a remaining ovary. During laparoscopic exploration, the mass was identified caudally distant from the right ovarian stump, adhered to the jejunal mesentery. The intestinal segment was apprehended and exteriorized by video-assisted technique, and removal of the tumor was performed associated with enterectomy of the affected segment. Evaluation of the left ovarian stump revealed a 0.5cm diameter cyst that was also removed. Histopathological analysis was suggestive of interstitial cell neoplasia (Leydig cell tumor) and an ovarian epithelial cyst. The Leydig cell tumor is frequently related in males, however, its appearance in bitches is considered uncommon, affecting mainly nonsterile animals. The mass non-adhered to the ovarian stump proves that remnants of ovarian tissue that may be free in the abdominal cavity and may be responsible for the development of the remaining ovary syndrome and neoplasms. The prognosis for ovarian neoplasms without metastasis is favorable, especially when surgical intervention is applied as a therapeutic method. The use of video surgery to assist in the diagnosis and treatment was considered appropriate for the case described, but it must be complemented by histopathological analysis.

Testicular morphometry of rats with Walker 256 tumor supplemented with L-glutamine

Abstract Glutamine is often used to treat metabolic changes associated with anorexia-cachexia syndrome in patients with malignant neoplasms. Walker 256 tumor is an excellent model for studying these changes associated with cancer in different organs, including injuries in testicular functions. However, the effects of supplementing glutamine on testicular morphometry in this model have not yet been investigated. Thus, the objective of this study was to evaluate the effect of L-glutamine supplementation on testicular morphometry in rats transplanted with Walker 256 tumor cells. Forty puberty Wistar rats were divided into four groups: control without L-glutamine (C); control supplemented with L-glutamine (CG); inoculated with Walker 256 tumor cells (WT) and inoculated with Walker 256 tumor cells and supplemented with L-glutamine (WTG). The testicles were removed, weighed, fixed in Bouin, and included in paraffin for histomorphometric analysis. Walker 256 tumor caused quantitative changes in the tubular and intertubular compartments and tunica albuginea, with reductions in the percentages of lumen and tunica albuginea, number of Sertoli cells per gram of testis; number of Leydig cells; percentage of blood vessels and connective tissue in intertubule. However, glutamine supplementation prevented part of these changes caused by the tumor, presenting mainly a protective effect on the tunica albuginea and percentage of blood and lymph vessels in the intertubule. These results indicate the potential of L-glutamine was able to recover for testicular dysfunction associated with cancer.

Testicular morphometry of rats with Walker 256 tumor supplemented with L-glutamine

Glutamine is often used to treat metabolic changes associated with anorexia-cachexia syndrome in patients with malignant neoplasms. Walker 256 tumor is an excellent model for studying these changes associated with cancer in different organs, including injuries in testicular functions. However, the effects of supplementing glutamine on testicular morphometry in this model have not yet been investigated. Thus, the objective of this study was to evaluate the effect of L-glutamine supplementation on testicular morphometry in rats transplanted with Walker 256 tumor cells. Forty puberty Wistar rats were divided into four groups: control without L-glutamine (C); control supplemented with L-glutamine (CG); inoculated with Walker 256 tumor cells (WT) and inoculated with Walker 256 tumor cells and supplemented with L-glutamine (WTG). The testicles were removed, weighed, fixed in Bouin, and included in paraffin for histomorphometric analysis. Walker 256 tumor caused quantitative changes in the tubular and intertubular compartments and tunica albuginea, with reductions in the percentages of lumen and tunica albuginea, number of Sertoli cells per gram of testis; number of Leydig cells; percentage of blood vessels and connective tissue in intertubule. However, glutamine supplementation prevented part of these changes caused by the tumor, presenting mainly a protective effect on the tunica albuginea and percentage of blood and lymph vessels in the intertubule. These results indicate the potential of L-glutamine was able to recover for testicular dysfunction associated with cancer.(AU)

Testicular tumors in 190 dogs: clinical, macroscopic and histopathological aspects / Neoplasmas testiculares em 190 cães: aspectos clínicos, macroscópicos e histopatológicos

This study aimed to characterize the prevalence and clinical, macroscopic and histopathological aspects of dogs affected by testicular tumors based on biopsy specimens from the Laboratório de Patologia Veterinária of the Universidade Federal de Santa Maria (LPV-UFSM) over 19 years. Parameters regarding the age, size, and breed of the affected dogs were also established. Of all dogs with some type of neoplasm submitted to histopathological analysis at the LPV over these 19 years (n=1,900), 213 (11.2%) had at least one testicular neoplasm. The tissues of 190 dogs (with 220 neoplasms) were available for histological reassessment. The dogs in this study had different types of testicular tumors with relatively similar frequencies. In descending order, the most frequent testicular neoplasms were seminomas (88/220), Leydig (interstitial) cell tumor (LCT; 64/220), Sertoli cell tumor (SCT; 61/220), and mixed germ cell-sex cord stromal tumor (MGSCT) (07/220). Among the dogs of defined breed (119 cases), large breeds had the largest number of cases (50/119), followed by small (47/119) and medium-sized (22/119) breeds. The ages of dogs affected by testicular tumors ranged from 10 months to 18 years. Increased testicular volume was the most common clinical manifestation. Eleven dogs presented information about clinical signs suggestive of hyperestrogenism syndrome (feminization). In seminomas, the diffuse pattern predominated over the intratubular pattern. Two sites (luminal and basal compartments) suggestive of the onset of neoplastic transformations in germ cells were observed in intratubular seminomas. They corroborate the hypothesis that canine seminomas possibly have pathogenesis similar to that observed in human spermatocytic seminomas. The SCTs and LCTs presented high cell morphology variation. SCTs had neoplastic cells organized in five different histological arrangements. As for LCT, solid-diffuse and cystic-vascular histological patterns were the most commonly observed. Through this study, it was possible to establish some of the leading clinical, macroscopic, and histopathological aspects of testicular neoplasms diagnosed over 19 years in the area covered by the LPV-UFSM.(AU)

Este estudo teve por objetivo caracterizar a prevalência, aspectos clínicos, macroscópicos e histopatológicos dos cães acometidos por neoplasmas testiculares, a partir dos espécimes de biópsias do Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV-UFSM) em 19 anos. Parâmetros quanto à idade, porte, raça dos cães acometidos também foram estabelecidos. De todos os cães com algum tipo de neoplasma submetido à análise histopatológica no LPV nesses 19 anos (n=1.900), 213 (11,2%) tinham ao menos um neoplasma testicular. Os tecidos de 190 cães (com 220 neoplasmas) estavam disponíveis para reavaliação histológica. Os cães deste estudo apresentaram diferentes tipos de neoplasmas testiculares com frequências relativamente semelhantes. Em ordem decrescente, os neoplasmas testiculares mais frequentes foram: seminomas (88/220), leydigomas (64/220), sertoliomas (61/220) e o tumor misto de células germinativas e do estroma do cordão sexual (MGSCT; 07/220). Dentre os cães com raça definida (119 casos), as raças de grande porte tiveram o maior número de casos (50/119), seguido das raças de pequeno (47/119) e médio porte (22/119). As idades dos cães acometidos por neoplasmas testiculares variaram de 10 meses a 18 anos. Aumento de volume testicular foi a manifestação clínica mais comum. Onze cães tinham informações sobre sinais clínicos sugestivos da síndrome da feminilização. Nos seminomas, houve o predomínio do padrão difuso sobre o intratubular. Dois locais (compartimentos luminal e basal) sugestivos de início das transformações neoplásicas nas células germinativas foram observados nos seminomas intratubulares, corroborando com a hipótese de que os seminomas caninos possivelmente tem patogênese semelhante à observada nos seminomas espermatocíticos humanos. Sertoliomas e leydigomas foram neoplasmas com alta variação na morfologia celular. Os sertoliomas tinham células neoplásicas dispostas em cinco arranjos histológicos distintos. Quanto aos leydigomas, os padrões histológicos sólido-difuso e cístico-vascular foram os mais comumente observados. Através deste estudo foi possível estabelecer alguns dos principais aspectos clínicos, macroscópicos e histopatológicos dos neoplasmas testiculares diagnosticados em 19 anos na área de abrangência do LPV-UFSM.(AU)

Testicular tumors in 190 dogs: clinical, macroscopic and histopathological aspects

ABSTRACT: This study aimed to characterize the prevalence and clinical, macroscopic and histopathological aspects of dogs affected by testicular tumors based on biopsy specimens from the Laboratório de Patologia Veterinária of the Universidade Federal de Santa Maria (LPV-UFSM) over 19 years. Parameters regarding the age, size, and breed of the affected dogs were also established. Of all dogs with some type of neoplasm submitted to histopathological analysis at the LPV over these 19 years (n=1,900), 213 (11.2%) had at least one testicular neoplasm. The tissues of 190 dogs (with 220 neoplasms) were available for histological reassessment. The dogs in this study had different types of testicular tumors with relatively similar frequencies. In descending order, the most frequent testicular neoplasms were seminomas (88/220), Leydig (interstitial) cell tumor (LCT; 64/220), Sertoli cell tumor (SCT; 61/220), and mixed germ cell-sex cord stromal tumor (MGSCT) (07/220). Among the dogs of defined breed (119 cases), large breeds had the largest number of cases (50/119), followed by small (47/119) and medium-sized (22/119) breeds. The ages of dogs affected by testicular tumors ranged from 10 months to 18 years. Increased testicular volume was the most common clinical manifestation. Eleven dogs presented information about clinical signs suggestive of hyperestrogenism syndrome (feminization). In seminomas, the diffuse pattern predominated over the intratubular pattern. Two sites (luminal and basal compartments) suggestive of the onset of neoplastic transformations in germ cells were observed in intratubular seminomas. They corroborate the hypothesis that canine seminomas possibly have pathogenesis similar to that observed in human spermatocytic seminomas. The SCTs and LCTs presented high cell morphology variation. SCTs had neoplastic cells organized in five different histological arrangements. As for LCT, solid-diffuse and cystic-vascular histological patterns were the most commonly observed. Through this study, it was possible to establish some of the leading clinical, macroscopic, and histopathological aspects of testicular neoplasms diagnosed over 19 years in the area covered by the LPV-UFSM.

RESUMO: Este estudo teve por objetivo caracterizar a prevalência, aspectos clínicos, macroscópicos e histopatológicos dos cães acometidos por neoplasmas testiculares, a partir dos espécimes de biópsias do Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV-UFSM) em 19 anos. Parâmetros quanto à idade, porte, raça dos cães acometidos também foram estabelecidos. De todos os cães com algum tipo de neoplasma submetido à análise histopatológica no LPV nesses 19 anos (n=1.900), 213 (11,2%) tinham ao menos um neoplasma testicular. Os tecidos de 190 cães (com 220 neoplasmas) estavam disponíveis para reavaliação histológica. Os cães deste estudo apresentaram diferentes tipos de neoplasmas testiculares com frequências relativamente semelhantes. Em ordem decrescente, os neoplasmas testiculares mais frequentes foram: seminomas (88/220), leydigomas (64/220), sertoliomas (61/220) e o tumor misto de células germinativas e do estroma do cordão sexual (MGSCT; 07/220). Dentre os cães com raça definida (119 casos), as raças de grande porte tiveram o maior número de casos (50/119), seguido das raças de pequeno (47/119) e médio porte (22/119). As idades dos cães acometidos por neoplasmas testiculares variaram de 10 meses a 18 anos. Aumento de volume testicular foi a manifestação clínica mais comum. Onze cães tinham informações sobre sinais clínicos sugestivos da síndrome da feminilização. Nos seminomas, houve o predomínio do padrão difuso sobre o intratubular. Dois locais (compartimentos luminal e basal) sugestivos de início das transformações neoplásicas nas células germinativas foram observados nos seminomas intratubulares, corroborando com a hipótese de que os seminomas caninos possivelmente tem patogênese semelhante à observada nos seminomas espermatocíticos humanos. Sertoliomas e leydigomas foram neoplasmas com alta variação na morfologia celular. Os sertoliomas tinham células neoplásicas dispostas em cinco arranjos histológicos distintos. Quanto aos leydigomas, os padrões histológicos sólido-difuso e cístico-vascular foram os mais comumente observados. Através deste estudo foi possível estabelecer alguns dos principais aspectos clínicos, macroscópicos e histopatológicos dos neoplasmas testiculares diagnosticados em 19 anos na área de abrangência do LPV-UFSM.

Leydigocitoma canino: aspectos ultrassonográficos, citológicos e histopatológicos / Canine Leydigocitoma: ultrasonographic, cytological and histopathological aspects

In non-neutered dogs, the testis is the second most common site for development of neoplasms, frequently occurring in elderly animals and with orchiectomy as main treatment. Regarding the great relevance of testicular disorders and the importance of accurate diagnoses, this study aims to report a case of canine leydigocytoma at the Veterinary Hospital in the State University of Ceará. The 10-years-old non-neutered Pitbull-breed animal was attended presenting hematuria and, physical examination showed a bilateral nodular formation in testicles in addition to an enlarged prostate. In ultrasonographic examination, the prostate was enlarged, presenting irregular shape and heterogeneous echotexture. These changes indicate the presence of prostatic hyperplasia. Both testicles presented well-defined hypoechogenic regions with evident marginal and internal vascularization, which indicate neoplastic neoformation. In cytology, a hypercellular epithelial sample was observed with cellular modifications suggestive of interstitial cell neoplasia. In the lab, macroscopically, the testis showed multiple soft, well-defined, white-colored to brownish nodules. Histopathology revealed neoplastic proliferation of Leydig cells, sustained by moderate fibrovascular stroma, with degenerated adjacent parenchyma and without signs of malignancy. Four months after orchiectomy, the prostate turned to normal dimensions, with no further ultrasonographic changes. This case reinforces the importance of bilateral orchiectomy and ultrasound for prostatic abnormalities and testicular neoplasms. The association of these conditions is not commonly described in the literature, being an important contribution in this regard.

Leydigocitoma canino: aspectos ultrassonográficos, citológicos e histopatológicos / Canine Leydigocitoma: ultrasonographic, cytological and histopathological aspects

In non-neutered dogs, the testis is the second most common site for development of neoplasms, frequently occurring in elderly animals and with orchiectomy as main treatment. Regarding the great relevance of testicular disorders and the importance of accurate diagnoses, this study aims to report a case of canine leydigocytoma at the Veterinary Hospital in the State University of Ceará. The 10-years-old non-neutered Pitbull-breed animal was attended presenting hematuria and, physical examination showed a bilateral nodular formation in testicles in addition to an enlarged prostate. In ultrasonographic examination, the prostate was enlarged, presenting irregular shape and heterogeneous echotexture. These changes indicate the presence of prostatic hyperplasia. Both testicles presented well-defined hypoechogenic regions with evident marginal and internal vascularization, which indicate neoplastic neoformation. In cytology, a hypercellular epithelial sample was observed with cellular modifications suggestive of interstitial cell neoplasia. In the lab, macroscopically, the testis showed multiple soft, well-defined, white-colored to brownish nodules. Histopathology revealed neoplastic proliferation of Leydig cells, sustained by moderate fibrovascular stroma, with degenerated adjacent parenchyma and without signs of malignancy. Four months after orchiectomy, the prostate turned to normal dimensions, with no further ultrasonographic changes. This case reinforces the importance of bilateral orchiectomy and ultrasound for prostatic abnormalities and testicular neoplasms. The association of these conditions is not commonly described in the literature, being an important contribution in this regard.(AU)

Effect of bacterial endotoxin lipopolysaccharide treatment on duck Leydig cells

This study aimed to investigate the effects of bacterial endotoxin lipopolysaccharide (LPS) on hormone production and gene expression in duck Leydig cells and its underlying mechanisms. Leydig cells were collected from 200-day-old mallard ducks and divided into five treatment groups (0, 50, 100, 200, and 400 ng/mL LPS). After treatment with LPS for 6, 12, 24, and 48 h, testosterone, activin, and inhibin levels in the cell supernatants were determined using enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of testosterone synthesis-related genes, including steroidogenic acute regulatory protein (StAR), 3-beta-hydroxysteroid dehydrogenase (3β-HSD), and cytochrome P450 aromatase (P450arom), and reproductive-related genes, including gonadotropin-inhibitory hormone receptor (GnIHR), follicle stimulating hormone receptor (FSHR), and luteinizing hormone receptor (LHR) were detected using quantitative real-time polymerase chain reaction (qRT-PCR). We successfully isolated and cultured duck Leydig cells with cell purity above 90%. Compared with the control group, the levels of testosterone, activin, and inhibin secreted in Leydig cells decreased gradually with increasing LPS concentration. After treatment with LPS, the expression of StAR and 3β-HSD genes in Leydig cells was upregulated at 12 h, and that of GnIHR was upregulated at 24 h; whereas the expression of FSHR and LHR was reduced at 24 h. This study indicates that LPS can inhibit the secretion of hormones and regulate the expression of related genes in duck Leydig cells.(AU)

Effect of bacterial endotoxin lipopolysaccharide treatment on duck Leydig cells

This study aimed to investigate the effects of bacterial endotoxin lipopolysaccharide (LPS) on hormone production and gene expression in duck Leydig cells and its underlying mechanisms. Leydig cells were collected from 200-day-old mallard ducks and divided into five treatment groups (0, 50, 100, 200, and 400 ng/mL LPS). After treatment with LPS for 6, 12, 24, and 48 h, testosterone, activin, and inhibin levels in the cell supernatants were determined using enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of testosterone synthesis-related genes, including steroidogenic acute regulatory protein (StAR), 3-beta-hydroxysteroid dehydrogenase (3β-HSD), and cytochrome P450 aromatase (P450arom), and reproductive-related genes, including gonadotropin-inhibitory hormone receptor (GnIHR), follicle stimulating hormone receptor (FSHR), and luteinizing hormone receptor (LHR) were detected using quantitative real-time polymerase chain reaction (qRT-PCR). We successfully isolated and cultured duck Leydig cells with cell purity above 90%. Compared with the control group, the levels of testosterone, activin, and inhibin secreted in Leydig cells decreased gradually with increasing LPS concentration. After treatment with LPS, the expression of StAR and 3β-HSD genes in Leydig cells was upregulated at 12 h, and that of GnIHR was upregulated at 24 h; whereas the expression of FSHR and LHR was reduced at 24 h. This study indicates that LPS can inhibit the secretion of hormones and regulate the expression of related genes in duck Leydig cells.

Testicular interstitial cell tumor with disseminated cutaneous metastasis in a dog

Background: Although testicular tumors commonly develop in the genital tract of dogs, only few studies have reportedabout metastatic testicular tumors in dogs to date. Therefore, the present study aimed to describe the gross, histopathological, and immunohistochemical findings in a dog with metastatic interstitial cell tumor.Case: A 6-year-old male Beagle underwent clinical examination due to enlargement of the left testicle and nodules inthe skin of the left inguinal region and of the scrotum. The left testicle was cryptorchid (inguinal), and the right testiclewas intrascrotal. Both testicles and affected skin fragments were surgically removed. The right testicle was normal insize and did not exhibit gross changes. However, the left testicular parenchyma had been completely replaced by a firm,grayish-to-white neoplastic mass with an irregular surface. The inguinal and scrotal skin fragments were thickened by theneoplastic masses. Histologically, the left testicular parenchyma was completely obliterated by a neoplastic mass consistingof round-to-oval and polygonal cells. The neoplastic cells generally had abundant eosinophilic cytoplasm with indistinctcell limits. The nuclei were round and small, with aggregated chromatin. However, some cells exhibited high degrees ofatypia and pleomorphism and large nuclei with granular chromatin. Mitotic figures were observed at high frequenciesof up to 11 per high-power field (400×). The mass in the left testicle was diagnosed as malignant interstitial cell tumor.The sections of the inguinal skin and scrotum presented with neoplastic cells similar to those observed in the left testicle,which is consistent with the metastasis of the malignant testicular interstitial cell tumor. On the basis of necropsy, multiple cutaneous and subcutaneous nodules were randomly distributed throughout the body, primarily in the prepuce, head,and limbs. Histological evaluations of the skin nodules...

Testicular interstitial cell tumor with disseminated cutaneous metastasis in a dog

Background: Although testicular tumors commonly develop in the genital tract of dogs, only few studies have reportedabout metastatic testicular tumors in dogs to date. Therefore, the present study aimed to describe the gross, histopathological, and immunohistochemical findings in a dog with metastatic interstitial cell tumor.Case: A 6-year-old male Beagle underwent clinical examination due to enlargement of the left testicle and nodules inthe skin of the left inguinal region and of the scrotum. The left testicle was cryptorchid (inguinal), and the right testiclewas intrascrotal. Both testicles and affected skin fragments were surgically removed. The right testicle was normal insize and did not exhibit gross changes. However, the left testicular parenchyma had been completely replaced by a firm,grayish-to-white neoplastic mass with an irregular surface. The inguinal and scrotal skin fragments were thickened by theneoplastic masses. Histologically, the left testicular parenchyma was completely obliterated by a neoplastic mass consistingof round-to-oval and polygonal cells. The neoplastic cells generally had abundant eosinophilic cytoplasm with indistinctcell limits. The nuclei were round and small, with aggregated chromatin. However, some cells exhibited high degrees ofatypia and pleomorphism and large nuclei with granular chromatin. Mitotic figures were observed at high frequenciesof up to 11 per high-power field (400×). The mass in the left testicle was diagnosed as malignant interstitial cell tumor.The sections of the inguinal skin and scrotum presented with neoplastic cells similar to those observed in the left testicle,which is consistent with the metastasis of the malignant testicular interstitial cell tumor. On the basis of necropsy, multiple cutaneous and subcutaneous nodules were randomly distributed throughout the body, primarily in the prepuce, head,and limbs. Histological evaluations of the skin nodules...(AU)

Seasonal influence on testicular morphophysiological parameters of bat Carollia perspicillata in fragments of the Atlantic Forest, northeastern Brazil / Influência sazonal sobre os parâmetros morfofisiológicos testiculares de Carollia perspicillata (Chiroptera: Phyllostomidae) em fragmentos florestais de Mata Atlântica, Nordeste do Brasil

Bats belong to the order Chiroptera, family Phyllostomidae, and present a wide diversity of reproductive strategies. However, information on the reproductive biology of male bats is scarce, mainly in the Northeast Region of Brazil. Thus, this study evaluated the seasonal testicular histomorphometry of the bat Carollia perspicillata in fragments of the Atlantic Forest in Pernambuco state. To this end, adult males were collected, euthanized for removal of the testicles, and later submitted to a routine histological technique. Histomorphometric analysis included assessment of the areas of tubular and intertubular compartment occupation, as well as quantification of spermatocytes, rounded spermatids, elongated spermatids, and Sertoli and Leydig cells. Results indicated that this bat species presents reproductive seasonality, because significantly higher averages of the testicular parameters were observed in the rainy season, which is a period of greater availability of food resources. Such inferences indicate that there is a synchrony between peak spermatogenesis and hormonal inversion in the months of high precipitation; furthermore, a higher carrying capacity of the Sertoli cells is noted. C. perspicillata males possibly present greater sperm and androgenic activity in the rainy season, associated with increased tubular area and number of spermatogenic cells, as well as with the intertubular area and number of Leydig cells, respectively.(AU)

Os morcegos pertencem a ordem Chiroptera, família Phyllostomidae, e apresentam ampla diversidade de estratégias reprodutivas. Entretanto, as informações relacionadas à biologia reprodutiva dos machos são escassas, principalmente no Nordeste do Brasil. Dessa forma, o trabalho avaliou a histomorfometria sazonal testicular de Carollia perspicillata em fragmentos de Mata Atlântica de Pernambuco. Para tanto, os machos adultos foram coletados, eutanasiados para a remoção dos testículos, e posteriormente submetidos à técnica histológica de rotina. As análises histomorfométricas avaliaram as áreas de ocupação do compartimento tubular e intertubular, assim como a quantificação dos espermatócitos, espermátides arredondadas, espermátides alongadas, células de Sertoli e de Leydig. Os resultados indicaram que a espécie apresenta sazonalidade reprodutiva, visto que maiores médias significativas dos parâmetros testiculares foram encontradas na estação chuvosa, que é um período de maior disponibilidade de recursos alimentares. Tais inferências indicam que existe uma sincronia entre o pico de espermatogênese e investimento hormonal nos meses de alta precipitação, atrelado a isso, nota-se ainda, uma maior capacidade de suporte das células de Sertoli. Os machos de C. perspicillata possivelmente apresentam uma maior atividade espermática e androgênica na estação chuvosa, associadas ao aumento da área tubular e do número de células espermatogênicas, assim como da área intertubular e do número das células de Leydig, respectivamente.(AU)