Cryopreservation of jaguar (Panthera onca) sperm cells using different cryoprotectants and different thawing temperatures

The cryopreservation of jaguar semen must be improved to produce high-quality biobanking doses. Until now, the rare studies of semen freezing in the species have only evaluated glycerol, always with a significant reduction in sperm quality in thawed semen. The purpose of this study was to assess the efficacy of three cryoprotectants, dimethylsulfoxide (DMSO), glycerol (GLY), and methanol (MET), in the cryopreservation of jaguar semen in an LDL-based extender, as well as the effect of thawing temperature on dosage quality. Five mature males with a history of reproduction were used. On the males, an infrared thermal image (IRT) was captured, the spicules and testes were analyzed, and the CASA system was used to evaluate the quality of fresh and thawed sperm. The superficial IRT was 4.6 ± 1.2 °C cooler than the anal sphincter, and the semen measured between 27.3 and 28.7 °C shortly after exiting the urethra. The total motility of fresh sperm was 55.3 ± 22.6%, and progressive motility was 36.3 ± 18%. The total motility of thawed sperm was 5.28 ± 2.51%, 4.49 ± %2.49, and 0.51 ± 0.62% for DMSO, GLY, and MET, respectively. DMSO and GLY performed better than MET, and there was no difference in thawing temperature (37°C 30 s vs. 50°C 12 s). All animals exhibit a considerable level of morphological changes in sperm. Low amounts of total and progressive motility were found in the thawed sperm. Males with a high level of sperm morphological changes were found to be fertile, but the lone male with normospermia was infertile. Thus, we contest the applicability of the commonly used morphological classification for bovines to felid species.(AU)

Penile fractures in young bulls raised in feedlots in Southern Brazil

Background: Penile fracture is a pathology of young cattle that perform precocious and disordered breeding. The incompatibility of height between males and females and sodomy between males cause a great pressure on the sigmoid flexure and retractor muscle of the penis, which are the main causes and sites of organ injury. This study aimed to describe the epidemiological and pathological aspects of penile fractures observed in young bulls raised in pre-export feedlots (PEFs) in southern Brazil. Cases: In 2 PEFs located in the municipalities of Pelotas (property 1) and Capão do Leão (property 2), 3 male cattle [1 from property 1 and 2 from property 2] presented subcutaneous edema in the foreskin and perineum, associated with dysuria. The evolution of the clinical picture was approximately 20 days in all cases, with evolution to death. The bovine necropsied on property 1 had an increased volume and inguinal edema, involving the penis and scrotal sac. Necrosis of the subcutaneous tissue and local musculature was also observed. The testicles were surrounded by the necrotic tissue, and the right testis was swollen, with flaccid parenchyma adhering to the tunica albuginea. In the necropsy of 1 bull from property 2, an increase in the inguinal volume was observed, with an extensive area of necrosis and edema extending from the prepuce to the caudal musculature of the scrotal sac. There were also marked varicosis in the sigmoid flexure and necrosis of the adjacent region, without the involvement of the corpus cavernosum. During the necropsy of the 2 young bulls, fragments of organs from the abdominal, thoracic, and brain cavities were collected and fixed in 10% buffered formalin. From the bull of the property 2, an anatomical piece consisting of the penis, prepuce, and testicles was also collected and fixed in 10% buffered formalin. After 48 h, the tissue samples were cleaved, embedded in paraffin, cut into 3-µm-thick sections, and stained using hematoxylin and eosin (HE). A histological evaluation of the penile lesions in both cattle revealed intense hemorrhage, congestion, and necrosis of the muscles and tissues adjacent to the corpus cavernosum. In addition to areas of dystrophic calcification, neutrophil and macrophage infiltration was also observed. In the bull from the property 1, an intense edema and proliferation of fibrous tissue surrounding the urethra were noted. There were also marked tubular degeneration and intense infiltration of neutrophils, lymphocytes, and macrophages in the inner portion of the tunica albuginea. Discussion: In the present cases, the diagnosis was based on epidemiological data associated with clinical signs and pathology. The macroscopic lesions observed were probably due to the involvement of blood vessels adjacent to the penis, which suffered trauma during sodomy mating among cattle. These lesions have been described in other reports of this pathology and in diseases, such as acropostitis-phimosis, fibropapilloma of the glans, preputial abscess, and urolithiasis, and the differential diagnosis of these diseases must be carried out, as they have different etiologies. In the bulls of the present study, no lesions were observed in the corpus cavernosum, and this condition was attributed to the presence of varicosis and accumulation of urine in the prepuce, due to the difficulty in exposing the penis. Histologically, there were intense hemorrhage, congestion, and necrosis of the muscles and tissues adjacent to the corpus cavernosum, with the infiltration of neutrophils and macrophages, and areas of dystrophic calcification. The presence of necrotic lesions in tissues adjacent to the penis may be related to hypoxia, vascular lesions, or the action of chemical elements present in the urine. In both cases, vascular lesions were present, which were attributed to the main triggering factor for the disease.

Anoftalmia bilateral em cão associada à malformação peniana congênita / Bilateral anophthalmia in dog associated with congenital penile malformation

Background: Anophthalmia refers to the complete absence of an eyeball. It is a very rare congenital condition that affects all breeds of dogs, which may be unilateral or bilateral. Anophthalmia results from a severe developmental deficiency in the primitive forebrain, prior to the formation of the optic sulcus. This condition has a complex etiology, and chromosomal, monogenic, and environmental causes have already been identified. There might be an association between anophthalmia and other congenital abnormalities. This case report aimed to report the case of a male dog with bilateral anophthalmia associated with penile malformation. Case: A 3-year-old male mongrel dog was evaluated in a clinical consultation at the Veterinary Medical Teaching Hospital of the Federal University of Piauí (UFPI) - Teresina - Brazil, with signs of anemia and diarrhea. His vaccinations were not up to date (multiple and anti-rabies), presented ectoparasites (ticks), and showed inappetence. The owner reported that the animal was blind from birth, was already well adapted to its environment, had the company of other animals, and had no access to the street. On clinical examination, the animal had a good body score. The eyelids were bilaterally present with normally developed eyelashes. Both the head and face had a symmetrical appearance, without structural abnormalities, and the palpebral fissures were narrow. When opening the eyelids manually, the presence of the conjunctiva was observed bilaterally, the orbits were shallow, and neither orbit contained the eyeball. Further inspection with an ophthalmic clinical flashlight did not reveal any other ocular structures. The initial diagnosis was suggestive of bilateral anophthalmia. Ultrasound examination of the ocular region confirmed the presence of bilateral anophthalmia. A general physical examination revealed a malformed penis with a persistent penile frenulum. Regarding systemic clinical signs (apathy, inappetence, diarrhea, and anemia), clinical and laboratory examinations led to the suspicion of verminosis. Discussion: The dog in this report had a good body score; however, this type of malformation can generate non-viable puppies. Anophthalmia is an extremely rare congenital ocular malformation, both in humans and companion animals, in which there are no ocular structures in the orbits, and these structures (orbits) in affected individuals are generally small and shallow, as seen in the dog in this report. Ultrasonographic examination confirmed the bilateral anophthalmic condition, being the most accurate method of diagnosing this congenital condition. The animal in this report presented with anophthalmia associated with penile malformation. This type of association until this case report has only been described in humans and mice, and is probably caused by a mutation in the SOX2 gene, which is responsible for the protein of the same name, which makes the maturation of ocular tissues in the primitive forebrain. Thus, it is likely that such congenital alterations are the cause of the malformations observed in this animal. Maternal vitamin A deficiency, a factor known to cause ocular malformations, may also be involved in the etiology of congenital diseases. Nutritional imbalances in dogs are common in the animal's region of origin The dog in this report had a normal life. Although it is a condition that causes blindness, dogs can have a normal life and welfare, even with bilateral anophthalmia.

Persistence of congenital penile frenulum in male cat: / Persistência de frênulo peniano congênito em gato macho:

The penile frenulum is a thin strip of connective tissue that connects the penis glans to the foreskin. Due to the few number of reports of this condition, this work reports the occurrence of persistent penile frenulum in a mixed breed domestic cat, of seven months old and 5.2 kg. In this particular case, penile frenulum was an incidental finding during clinical examination. The animal was in puberty, characterizing the penile frenulum. The resolution of the case occurred spontaneously prior to elective orchiectomy. Thus, the changes observed in the penis due to congenital alteration in cats are described in this report.

O frênulo peniano é uma fina faixa de tecido conectivo que une a glânde ao prepúcio que pode se romper sob a ação da testosterona quando o animal inicia a puberdade. Devido a escassez de relatos desta afecção o presente trabalho relata a ocorrência de frênulo peniano persistente em um gato doméstico, sem raça definida, de sete meses de idade e 5,2 kg. O frênulo peniano foi um achado incidental durante o exame clínico do animal. O animal já apresentavase em puberdade, caracterizando o quadro de frênulo peniano. A resolução do caso ocorreu de maneira espontânea previamente a orquiectomia eletiva. Dessa forma, as alterações observadas no pênis devido a alteração congênita em gatos são descritas neste relato.

Parafimose em touro com lesão da extremidade livre do pênis como intercorrência da enfermidade acropostite relato de caso / Paraphimosis in bull with lesion of the free end of the penis as a complication of acropostite disease - case report

As particularidades morfológicas e anatômicas de touros podem predispor à ocorrência de enfermidades acometendo a genitália externa, dentre as quais a acropostite é considerada a de maior importância. O estudo relata um caso de acropostite em que o animal, além dos sinais clínicos normais da enfermidade, apresentou exposição peniana permanente, característico de parafimose. Informações sobre a epidemiologia, fatores de risco, diagnóstico e tratamento foram apresentados. Fragmentos das áreas lesionadas foram colhidos e encaminhados para exame histopatológico. Verificou-se hiperplasia pseudocarcinomatosa acentuada da camada espinhosa associada a crostas serocelulares superficiais. A técnica cirúrgica mostrou-se eficiente e passível de ser realizada a campo. (AU)

The morphological and anatomical characteristics of bulls may predispose to the occurrence of diseases affecting the external genitalia, among which acropostite is considered the most important. The study reports a case of acropostite where the animal, beyond the normal clinical signs of the disease, presented permanent penile exposure, characteristic of paraphimosis. Information on the epidemiology, risk factors, diagnosis and treatment were presented. Fragments of injured areas were collected and sent for histopathological examination. It was verified pronounced pseudocarcinomatous hyperplasia on superficial spinous layer with serocelulares crusts. The surgical technique was efficient and able to be carried out in the field. (AU)

Parafimose em touro com lesão da extremidade livre do pênis como intercorrência da enfermidade acropostite – relato de caso / Paraphimosis in bull with lesion of the free end of the penis as a complication of acropostite disease - case report

As particularidades morfológicas e anatômicas de touros podem predispor à ocorrência de enfermidades acometendo a genitália externa, dentre as quais a acropostite é considerada a de maior importância. O estudo relata um caso de acropostite em que o animal, além dos sinais clínicos normais da enfermidade, apresentou exposição peniana permanente, característico de parafimose. Informações sobre a epidemiologia, fatores de risco, diagnóstico e tratamento foram apresentados. Fragmentos das áreas lesionadas foram colhidos e encaminhados para exame histopatológico. Verificou-se hiperplasia pseudocarcinomatosa acentuada da camada espinhosa associada a crostas serocelulares superficiais. A técnica cirúrgica mostrou-se eficiente e passível de ser realizada a campo.

The morphological and anatomical characteristics of bulls may predispose to the occurrence of diseases affecting the external genitalia, among which acropostite is considered the most important. The study reports a case of acropostite where the animal, beyond the normal clinical signs of the disease, presented permanent penile exposure, characteristic of paraphimosis. Information on the epidemiology, risk factors, diagnosis and treatment were presented. Fragments of injured areas were collected and sent for histopathological examination. It was verified pronounced pseudocarcinomatous hyperplasia on superficial spinous layer with serocelulares crusts. The surgical technique was efficient and able to be carried out in the field.

BLOQUEIOS LOCORREGIONAIS EM BUBALINOS: BLOQUEIO DO NERVO PUDENDO, PARAVERTEBRAL E TORÁCICO LATERAL

Com o objetivo de avaliar os efeitos dos bloqueios locorregionais: do nervo pudendo nos machos e paravertebral isolado e associado ao torácico lateral nas fêmeas, usando lidocaína 2% com vasoconstritor, foram utilizados 25 búfalos, sendo 13 machos, divididos em dois grupos (adultos e jovens) e 12 fêmeas. Observou-se a ocorrência ou não de bloqueio motor (BM), por meio da inspeção direta do relaxamento da musculatura, nos machos; para verificar o bloqueio sensitivo (BS) foi utilizada agulha hipodérmica nos machos e pinça dente de rato nas fêmeas no intuito de buscar respostas à dor. O período de bloqueio motor e sensitivo foi determinado a partir da apresentação de relaxamento e não reação aos estímulos dolorosos, avaliados a cada 20 minutos nos machos e a cada 10 minutos nas fêmeas até retorno do vigor e dor. Em machos no GJ e GA respectivamente, ocorreu a exposição do pênis em 83% e 85.7%, iniciaram 20.2±17 min. e 19.6±10.1min., durando 91.6±84.2 min. e 110±179.6 min. O BS do pênis não ocorreu em nenhum grupo. O BM do escroto ocorreu em 100% em ambos os grupos, iniciou 10.8±4.9 min e 9.8±5.6 min., durando 138.3±51.9 min. e 139.2±83.5 min., no GJ e GA, respectivamente. O BS do escroto ocorreu de forma U ou B em 83.2% e 100%, iniciou 11.5±7.7 min. e 17.5±7.2 min., durando 103.8±80.9 min. e 108.3±94.9 min., no GJ e GA, respectivamente. Nas fêmeas ambos os bloqueios foram eficientes em anestesiar as regiões do flanco e região ventral do abdômen. A anestesia ocorreu em 83% e 91.3% na fossa paralombar (FPL), de 91.3% e 91.3% na lateral (L) e 83% e 91.3% na região ventral (RV), nos bloqueios paravertebral (BPV) e paravertebral associado ao torácico lateral (BPV+BTL) respectivamente. Porém, ambos os bloqueios não foram eficientes em anestesiar os tetos anteriores (TA), a parte caudal do úbere (UCl) e os tetos posteriores (TP), anestesiando apenas 16.6% dos animais. Foi na porção cranial do úbere (UCr) que ocorreu a maior diferenças entres os bloqueios, sendo 24.9% no BPV e 58.1% no (BPV+BTL). Desta forma, verificou-se a eficiência das técnicas para exposição peniana e anestesia na região cranial do úbere.

In order to evaluate the effects of locoregional anesthesia blocks of the pudendal nerve in males, and isolated paravertebral and associated with the lateral thoracic in females, using 2% lidocaine with vasoconstrictor, 25 buffaloes were used, 13 males, divided into two groups (adults and young) and 12 females. Motor block (BM) was observed if present or not, through direct inspection of muscle relaxation in males. To check the sensory block (BS), a hypodermic needle was used in males and an Adson forceps in females in order to seek answers to pain. Motor block (BM) was observed if present not, through direct inspection of muscle relaxation in males; to check the sensory block (BS), a hypodermic needle was used in males and an Adson forceps in females in order to seek response to pain. The period of motor and sensory block was determined from the presentation of relaxation and non-reaction to painful stimuli, assessed every 20 minutes in males and every 10 minutes in females until the return of vigor and pain. In males in GJ and GA respectively, penis exposure occurred in 83% and 85.7%, starting at 20.2 ± 17 min. and 19.6 ± 10.1 min., lasting 91.6 ± 84.2 min. and 110 ± 179.6 min. The BS of the penis did not occur in any group. The scrotum BM occurred in 100% in both groups, starting at 10.8 ± 4.9 min and 9.8 ± 5.6 min., lasting 138.3 ± 51.9 min. and 139.2 ± 83.5 min., in GJ and GA, respectively. The scrotum BS occurred in a U or B form in 83.2% and 100%, starting at 11.5 ± 7.7 min. and at 17.5 ± 7.2 min., lasting 103.8 ± 80.9 min. and 108.3 ± 94.9 min., in GJ and GA, respectively. In females, both blocks were efficient in anesthetizing the flank and ventral regions of the abdomen. Anesthesia occurred in 83% and 91.3% in the paralumbar fossae (FPL), 91.3% and 91.3% in the lateral (L) and 83% and 91.3% in the ventral region (RV), in paravertebral (BPV) and paravertebral blocks associated with lateral thoracic (BPV + BTL) respectively. However, both blocks were not efficient in anesthetizing the anterior teat (TA), the caudal part of the udder (UCl) and the posterior teat (TP), anesthetizing only 16.6% of the animals. It was in the cranial portion of the udder (UCr) that the major differences occurred between the blocks, with 24.9% in BPV and 58.1% in (BPV + BTL). Thus, the efficiency of techniques for penile exposure and anesthesia in the cranial region of the udder was verified.

Diagnóstico clínico, laboratorial e tratamento cirúrgico do carcinoma de células escamosas no genital de equinos machos: relatos de dois casos / Clinical and laboratorial diagnosis and surgical treatment of genitalsquamous cell carcinoma in male horses: report of two cases

O carcinoma de células escamosas (CCE) tem importância relevante na rotina clínica e cirúrgica de equinos e o diagnóstico dessa neoplasia é baseado nos exames clínico e histopatológico. Como a terapia conservativa raramente apresenta resultados satisfatórios,a intervenção cirúrgica tem-se mostrado mais vantajosa, constituindo, em algumas situações, como única alternativa para evitar o comprometimento morfofuncional da estrutura anatômica ou mesmo o óbito do paciente. Este estudo objetivou estabelecer o diagnóstico clínico e laboratorial de CCE genital em equinos, descrever o tratamento cirúrgico empregando sutura captonada como alteração da técnica convencional e avaliar os procedimentos pós-operatórios. Foram utilizados dois animais do sexo masculino,com cerca de 20 anos, um mestiço Apaloosa e outro pônei. Após o diagnóstico clínico foram submetidos ao tratamento cirúrgico mediante anestesia geral e colheita de material para exames citológico e histopatológico. O resultado do exame histopatológico confirmou a suspeita diagnóstica de CCE. Concluiu-se que os exames clínicos e histopatológicos são fundamentais para se estabelecer o diagnóstico do CCE genital em equinos do sexo masculino, porém, para evitar dois procedimentos anestésicos subsequentes e minimizar possíveis complicações decorrentes da anestesia, a colheita de material para avaliação laboratorial deve ser realizada durante o tratamento cirúrgico.(AU)

Squamous cell carcinoma (SCC) has relevant importance in the clinical and surgery of equines and the diagnosis is based on clinical and histopathological exams. The conservative therapy rarely gives satisfactory results while the surgical intervention is more advantageous, constituting, in some situations, the only alternative to avoid compromising anatomical structure or eventhe death of the patient. This study aimed to establish the clinical and laboratory diagnosis of genital SCC, describing the quilled suture as changing the conventional technique and evaluating post operative procedures. Were used two male animals, about 20 years old, a half-breed appaloosa and other pony. After the clinical diagnosis, underwent surgery under general anesthesia and collection of material for histopathological examination. The results of this examination confirmed the suspected diagnosis. It was concluded that the clinical and histopathological tests are essential to establish the diagnosis of genital SCC in male horses, however. To avoid two subsequent anesthesia and minimize possible complications during the proceeding, material for laboratoryevaluation should be collected during surgery.(AU)

Diagnóstico clínico, laboratorial e tratamento cirúrgico do carcinoma de células escamosas no genital de equinos machos: relatos de dois casos / Clinical and laboratorial diagnosis and surgical treatment of genitalsquamous cell carcinoma in male horses: report of two cases

O carcinoma de células escamosas (CCE) tem importância relevante na rotina clínica e cirúrgica de equinos e o diagnóstico dessa neoplasia é baseado nos exames clínico e histopatológico. Como a terapia conservativa raramente apresenta resultados satisfatórios,a intervenção cirúrgica tem-se mostrado mais vantajosa, constituindo, em algumas situações, como única alternativa para evitar o comprometimento morfofuncional da estrutura anatômica ou mesmo o óbito do paciente. Este estudo objetivou estabelecer o diagnóstico clínico e laboratorial de CCE genital em equinos, descrever o tratamento cirúrgico empregando sutura captonada como alteração da técnica convencional e avaliar os procedimentos pós-operatórios. Foram utilizados dois animais do sexo masculino,com cerca de 20 anos, um mestiço Apaloosa e outro pônei. Após o diagnóstico clínico foram submetidos ao tratamento cirúrgico mediante anestesia geral e colheita de material para exames citológico e histopatológico. O resultado do exame histopatológico confirmou a suspeita diagnóstica de CCE. Concluiu-se que os exames clínicos e histopatológicos são fundamentais para se estabelecer o diagnóstico do CCE genital em equinos do sexo masculino, porém, para evitar dois procedimentos anestésicos subsequentes e minimizar possíveis complicações decorrentes da anestesia, a colheita de material para avaliação laboratorial deve ser realizada durante o tratamento cirúrgico.

Squamous cell carcinoma (SCC) has relevant importance in the clinical and surgery of equines and the diagnosis is based on clinical and histopathological exams. The conservative therapy rarely gives satisfactory results while the surgical intervention is more advantageous, constituting, in some situations, the only alternative to avoid compromising anatomical structure or eventhe death of the patient. This study aimed to establish the clinical and laboratory diagnosis of genital SCC, describing the quilled suture as changing the conventional technique and evaluating post operative procedures. Were used two male animals, about 20 years old, a half-breed appaloosa and other pony. After the clinical diagnosis, underwent surgery under general anesthesia and collection of material for histopathological examination. The results of this examination confirmed the suspected diagnosis. It was concluded that the clinical and histopathological tests are essential to establish the diagnosis of genital SCC in male horses, however. To avoid two subsequent anesthesia and minimize possible complications during the proceeding, material for laboratoryevaluation should be collected during surgery.

Penectomy and scrotal urethrostomy in dogs: report of four cases / Penectomia com uretrostomia escrotal em cães: relato de quatro casos

Wide trauma and neoplasms constitute the main indications for penectomy and scrotal urethrostomy in the dog. Reconstruction of the prepuce is technically challenging, which may not be feasible in cases of extensive tissue loss, requiring penile amputation. The aim of the current study was to report four cases of penectomy associated to scrotal urethrostomy in dogs. The patients were subjected to hematological, histopathologic exams, thoracic radiographs and abdominal echographic evaluations for surgical triage. Malignant neoplasms were diagnosed in three patients and fibrosis with extensive penile exposition was found in one dog. The patients were submitted to penile amputation and scrotal urethrostomy postoperative complications included mild hemorrhage in one dog and wound infection and myiasis in another dog. In conclusion, surgical resection followed by penectomy with scrotal urethrostomy was the best option for the treatment of the preputial lesions and resolution was achieved.

Os traumatismos extensos de prepúcio e pênis, além de neoplasias, constituem as principais indicações cirúrgicas de penectomia associado à uretrostomia escrotal no cão. A reconstrução do prepúcio é desafiadora e torna-se inviável em casos de lesões amplas, requerendo a amputação peniana. O objetivo do presente estudo foi relatar quatro casos de indicação para penectomia associada à uretrostomia escrotal em cães. Os pacientes foram submetidos a exames complementares hematológicos, citologia das lesões, radiografia de tórax, ultrassonografia abdominal foram realizadas para triagem cirúrgica. Em três pacientes diagnosticou-se neoplasias malignas e em um paciente, fibrose com extensa exposição peniana. Os cães foram submetidos a penectomia, associado a uretrostomia escrotal. As complicações pós-operatórias incluíram hemorragia leve em um paciente e infecção e miiase em outro cão. Concluiu-se que a ressecção cirúrgica seguida de penectomia com uretrostomia escrotal foi a melhor opção para as lesões prepuciais, obtendo-se sucesso no tratamento realizado.

Penectomy and scrotal urethrostomy in dogs: report of four cases / Penectomia com uretrostomia escrotal em cães: relato de quatro casos

Wide trauma and neoplasms constitute the main indications for penectomy and scrotal urethrostomy in the dog. Reconstruction of the prepuce is technically challenging, which may not be feasible in cases of extensive tissue loss, requiring penile amputation. The aim of the current study was to report four cases of penectomy associated to scrotal urethrostomy in dogs. The patients were subjected to hematological, histopathologic exams, thoracic radiographs and abdominal echographic evaluations for surgical triage. Malignant neoplasms were diagnosed in three patients and fibrosis with extensive penile exposition was found in one dog. The patients were submitted to penile amputation and scrotal urethrostomy postoperative complications included mild hemorrhage in one dog and wound infection and myiasis in another dog. In conclusion, surgical resection followed by penectomy with scrotal urethrostomy was the best option for the treatment of the preputial lesions and resolution was achieved.(AU)

Os traumatismos extensos de prepúcio e pênis, além de neoplasias, constituem as principais indicações cirúrgicas de penectomia associado à uretrostomia escrotal no cão. A reconstrução do prepúcio é desafiadora e torna-se inviável em casos de lesões amplas, requerendo a amputação peniana. O objetivo do presente estudo foi relatar quatro casos de indicação para penectomia associada à uretrostomia escrotal em cães. Os pacientes foram submetidos a exames complementares hematológicos, citologia das lesões, radiografia de tórax, ultrassonografia abdominal foram realizadas para triagem cirúrgica. Em três pacientes diagnosticou-se neoplasias malignas e em um paciente, fibrose com extensa exposição peniana. Os cães foram submetidos a penectomia, associado a uretrostomia escrotal. As complicações pós-operatórias incluíram hemorragia leve em um paciente e infecção e miiase em outro cão. Concluiu-se que a ressecção cirúrgica seguida de penectomia com uretrostomia escrotal foi a melhor opção para as lesões prepuciais, obtendo-se sucesso no tratamento realizado.(AU)

Avaliação de dois diluidores para a criopreservação do sêmen de mico-leão-de-cara-dourada (Leontopithecus chrysomelas).

Muitas espécies de primatas neotropicais são ameaçadas de extinção. As biotecnologias da reprodução podem contribuir consistentemente para a conservação das espécies, estando entre elas os bancos de recurso genético. Entretanto, o sucesso no uso de tais tecnologias depende de avanços no conhecimento da biologia reprodutiva básica de uma certa espécie. Durante esse processo, a avaliação da qualidade espermática é de suprema importância. Nós concentramos nosso trabalho em micos-leões-de-cara-dourada (Leontopithecus chrysomelas), mantidos na Fundação Parque Zoológico de São Paulo. Leontopithecus sp. é um alvo importante para o desenvolvimento de técnicas reprodutivas com o propósito da conservação, sendo que todas as espécies do gênero são classificadas como ameaçadas ou criticamente ameaçadas de extinção pela IUCN. Para a avaliação da qualidade espermática em L. chrysomelas, técnicas como o teste de ligação do espermatozoide, e as colorações não fluorescentes para a avaliação de integridade de acrossomo fast-green/rosa bengala, integridade de membrana plasmática eosina-nigrosina e atividade mitocondrial 3,3 diaminobenzidina foram validados para sêmen fresco. Nós também validamos uma técnica de avalição de integridade de acrossomo corante azul de coomassie, para amostras descongeladas devido a inadequação do método utilizado em sêmen fresco. Algumas espécies de primatas neotropicais apresentam variação na qualidade seminal ao longo do ano. Para avaliar a qualidade do sêmen para essa espécies e para verificar se mico-leão-de-cara-dourada possui essa variação, nós colhemos sêmen nas estações de seca e chuva, durante diferentes períodos da estação reprodutiva no intervalo dos períodos conceptivos e no final do segundo período conceptivo. As medições dos testículos foram realizadas previamente a cada colheita de sêmen, e uma diferença significativa foi encontrada no volume testicular total entre as estações, com maiores volumes encontrados durante a estação de seca (p = 0,0011). As amostras colhidas durante a estação de chuva mostraram uma maior porcentagem de motilidade total e integridade de membrana plasmática (p = 0,0149 e p = 0,0279, respectivamente), durante a estação de chuva. Mesmo com essas diferenças, sêmen de ambas as estações apresentaram boa qualidade. A taxa de sucesso das colheitas com a técnica de vibroestimulação peniana foi de 100%, e todas as amostras eram constituídas de somente uma fração coagulada. Apesar de não obtermos a dissolução completa do coágulo, nós pudemos acessar um grande número de espermatozóides pela adição do meio Biggers-Whitten-Whittingham (BWW) ao coágulo por 30 minutos a 37C, e realizamos todas as análises propostas. As mesmas amostras usadas nas avaliações do sêmen fresco foram subsequentemente submetidas à criopreservação usando dois diluidores comerciais diferentes BotuBOV (BB) e Freezing Medium with Glycerol and Gentamicin Test Yolk Buffer (TYB). As amostras descongeladas foram analisadas com os mesmos métodos previamente validados e também foram avaliadas para a suscetibilidade ao estresse oxidativo pela quantificação de substâncias reativas ao ácido tiobarbitúrico (TBARS). Para todos os parâmetros seminais avaliados, não houveram diferenças significativas entre BB e TYB, exceto que o BB apresentou porcentagem de acrossomos íntegros (p = 0.0101) e concentração de TBARS (p = 0.0005) maiores. Apesar dessas diferenças, as amostras descongeladas de ambos diluidores mostraram resultados similares no teste de predição da capacidade fertilizante (ensaio de ligação do espermatozoide). Do nosso trabalho, podemos observar que os métodos propostos foram validados com sucesso para L. chrysomelas. A qualidade do sêmen fresco foi muito alta e ambos diluidores obtiveram sucesso na criopreservação do sêmen.

Many Neotropical primate species are endangered of extinction. Reproductive biotechnologies can consistently contribute to species conservation, being among them the genetic resource banks. However, success in the use of such technologies relies on advances in the knowledge of basic reproductive biology of a given species. During these processes, assessment of sperm quality is of supreme importance. We focused our work in captive golden-headed lion tamarins (Leontopithecus chrysomelas), housed at São Paulo Zoological Park Foundation. Leontopithecus sp. is a very important target for the development of reproductive techniques aiming conservation, as all species from this genus are classified as endangered or critically endangered of extinction by IUCN. For the evaluation of sperm quality in L. chrysomelas, techniques like sperm-binding assay, and the non-fluorescent staining for evaluation of acrosome integrity fast-green/rose-bengal, plasma membrane integrity eosin-nigrosin and mitochondrial activity 3.3 diaminobenzidine were validated for fresh semen. We have also validated an acrosome integrity evaluation technique coomassie blue staining, for thawed samples due to the inadequacy of the method used for fresh semen. Some Neotropical primates species show variation in semen quality over the year. In order to assess semen quality for this species and to verify if golden-headed lion tamarin presents this variation, we collected semen in dry and rainy seasons, during different periods of breeding season in the interval of conceptive periods and in the end of the second conceptive period. Testicular measurements were taken prior each semen collection, and a significant difference was found in total testicular volume between seasons, with major volumes found during dry season (p = 0.0011). Samples collected during rainy season showed higher percentage of total motility and intact plasma membrane (p = 0.0149 and p = 0.0279, respectively), during rainy season. Even with those differences, semen from both seasons presented good quality. The success rate for collection with penile vibrostimulation technique was of 100%, and all samples obtained were constituted by only a coagulated fraction. Despite not achieving coagulum dissolution, we were able to access a high number of spermatozoa by adding Biggers-Whitten-Whittingham (BWW) medium to the coagulum for 30 min at 37C, and performed all proposed analyzes. The same samples used of fresh analyzes were subsequently submitted to cryopreservation using two different commercial extenders BotuBOV (BB) and Freezing Medium with Glycerol and Gentamicin Test Yolk Buffer (TYB). Thawed semen samples were analyzed with the same methods previously validated and were also evaluated for the susceptibility to oxidative stress through quantification of induced thiobartituric acid reactive substances (TBARS). For all seminal parameters assessed, no significant difference was observed between BB and TYB, except that BB presented a higher percentage of intact acrosome (p = 0.0101) and concentration of TBARS (p = 0.0005). Despite those differences, thawed semen from both extenders performed similarly at the prediction of fertilizing capacity (sperm-binding assay). From our work, we observed that the proposed methods were successfully validated for L. chrysomelas. Fresh semen quality was very high and both extenders were successful in cryopreserving semen.

CARACTERÍSTICAS SEMINAIS DO SAGUI-DA-SERRA-ESCURO (CALLITHRIX AURITA, GEOFFROY IN HUMBOLDT, 1812) (PRIMATES, CALLITRICHIDAE)

O sagui-da-serra-escuro (Callithrix aurita) é uma espécie de ocorrência das florestas tropicais do sudeste do Brasil e encontra-se classificada como em perigo na Lista Nacional Oficial de Espécies da Fauna Ameaçadas de Extinção, sendo pouco estudada até os dias atuais. A colheita e análise do sêmen é uma das formas essenciais de avaliação do potencial reprodutivo de machos. Foram utilizados quatro machos, que se encontravam em cativeiro no Centro de Primatologia do Rio de Janeiro (CPRJ), sendo estes dois machos adaptados ao cativeiro, e dois resgatados e alojados no início do experimento para descrever a biometria testicular e determinar um protocolo de colheita e avaliação seminal. Utilizou-se um tubo de vidro cilíndrico, no qual o pênis foi inserido para a estimulação vibratória e colheita do sêmen. Para biometria testicular o valor médio de volume testicular total (VTT) foi de 941,4±340,0mm3 , com variação de 339,61743,6 mm3 (valor mínimo e máximo). Obtiveram-se 20 ejaculações com sucesso das 39 tentativas realizadas (51,3%). A maioria das ejaculações ocorreu na frequência 80 Hz e amplitude 1,0 mm. A média do volume obtido foi 10,48±7 µL entre os animais. Após a colheita do material, realizaram-se as avaliações macroscópicas e microscópicas Foram encontradas médias de motilidade 37,89±30%, concentração espermática total 14,62± 6 x106 sptz/mL, 29,7±8,1% de espermatozóides normais, defeitos maiores 17,0±8,5%, defeitos menores 50,8±12,9%, integridade funcional da membrana plasmática 90,9±5,8% e integridade da membrana plasmática 53,32±23%. A técnica de vibro estimulação consistiu em um método exequível para colheita de sêmen em Callithrix aurita. Animais adaptados apresentaram maior eficiência nas ejaculações, mostrando que a adaptação ao ambiente em cativeiro favorece a eficiência e manipulação dos animais, e obtenção das amostras seminais. Foi observada uma variação de resultados das características seminais ao longo do ano do experimento, e ainda, com associação entre os parâmetros analisados como VTT, volume do ejaculado e concentração espermática total.

The Buffy-tufted marmoset (Callithrix aurita) has a occurrence in tropical forests of southeastern Brazil and is classified as "endangered" in the National Official Species of Endangered Fauna and have been poorly investigated until the current days. The collection and analysis of semen is one of the essential ways of assessing the reproductive potential of males. Four males were used, which were in captivity at the Primate Center of Rio de Janeiro (CPRJ). Two males were adapted to captivity, and two rescued and housed at the beginning of the experiment to describe the testis size and determine a collecting protocol and seminal evaluation. We used a cylindrical glass tube in which the penis is inserted for vibratory stimulation and collection of semen. For testis size the average value of total testicular volume (VTT) was 941.4 ± 340.0mm3 , ranging from 339.6 to 1743.6 mm3 (minimum and maximum). There was obtained 20 successful ejaculations of 39 attempts (51.3%). Most ejaculation occurred in the frequency range 80 Hz and 1.0 mm. The average of the obtained volume was 10.48 ± 7 uL between animals. After the collection of the material, there were macroscopic and microscopic evaluations that provided data of the seminal characteristics of the species in captivity. In sperm assessments were found 37.89 ± 30% average motility, 14.62 ± 6 x106 sperm / ml of total sperm concentration 29,7±8,1% of normal sperm, 17,0±8,5% major defects 50,8±12,9% minor defects, 90.9 ± 5.8% functional membrane integrity and 53.32 ± 23% plasma membrane integrity. The vibro stimulation technique consists of a feasible method for semen collection in buffy-tufted marmoset. Adapted animals showed higher efficiency in ejaculations, showing that adaptation to captive environment favors efficiency, animal manipulation and obtaining semen samples. A variation of results of seminal characteristics throughout the year of the experiment was observed, and also with the association between the parameters analyzed as VTT, the ejaculate volume and total sperm concentration.

Avaliação dos gânglios regionais no carcinoma epidermóide do pênis (cep) através de um índice de risco

The aim of this study was to investigate the predictive value of a risk index (RI) based on tumor pT stage and grade on the prognosis for inguinal lymphactic metastasis. We included 53 patients bearing schamous cell carcinoma of the penis (SCCP) treated by penectomy at the Hospital das Clínicas da FMRP-USP from 1978 to 1995. Age range varied from 32 to 97 years. Tumor grading was determined according to Broders method and staging by 1999 TNM system. RI was calculated by the sum of pT stage and grade (RI = pT+G). Bilateral inguinal lymphdenectomy was carried out within 6 weeks the penectomy in 14 patients cN+ at the presentation. The remaining 36 cases were kept under surveillance. In 9 of these patients required a delayed regional lymphadenectomy due to nodal relapse (median follow-up: 9 months). Proportions of pN0 patients according to RI were: 92% - 2, 80% - 3 and 0% - 6. The proportion of pN+ patients with a pooled RI of 4, 5 and 6 was significantly higher than that seen in pooled cases with lower RI (p = 0,004). From 36 patients initially cN0, 27 were RI 2 or 3 ( 5 of them progressed later on to pN+ , 19%) and 9 were RI ³ 4 (3 of them had progression to lymphatic spread, 33%). But, the difference observed in cN0 patients was not significant (p = 0,3). In conclusion, patients with RI ³ 4 were at higher risk of nodal spread when the overall sample was at stake, but this cutoff level for RI had no predictive value for the patients that were initially cN0.

O objetivo desse estudo foi investigar a validade de um índice de risco (IR) no prognóstico de metástases ganglionares no carcinoma epidermóide do pênis (CEP). Foram analisados 53 pacientes com CEP atendidos no Hospital das Clínicas da FMRP-USP, de janeiro de 1978 a dezembro de 1995. A idade variou de 32 a 97 anos. Os tumores foram graduados de acordo com a classificação de Broders e estadiados retrospectivamente (TNM 1999). O IR foi determinado pela soma de TeG (IR = T+G). A linfadenectomia inguinal bilateral (LD) foi indicada naqueles que apresentaram gânglios suspeitos após a antibioticoterapia ou que vieram apresentar alterações durante o período de observação. Faleceram 17 pacientes; A LD foi realizada em 14 pacientes e os 39 restantes permaneceram em observação clínica, sendo que em 8 deles foi feito LD após tempo mediano de 9 meses. Dos pacientes com índice de risco 2 e 3 92% e 80% respectivamente não apresentavam metastase ganglionar, ao passo que a totalidade daqueles com IR=6 tinha gânglios comprometidos. A associação dos grupos 2 e 3, comparada ao conjunto 4, 5 e 6, mostrou percentual maior de comprometimento nestes últimos (p = 0,0046). Dos 36 pacientes inicialmente No, 27 apresentavam IR 2 ou 3, dos quais 5 evoluíram para N+ (19%); os 9 restantes eram IR > 4 e 3 também evoluíram para N+ (33%). Apesar da diferença, não houve significância estatística entre eles (p = 0,3837). Embora tenha ocorrido maior comprometimento naqueles com IR > 4, o índice de risco não foi capaz de identificar previamente pacientes que evoluíram para N+.

CD34 immunoexpression in penile carcinoma

OBJECTIVE: To investigate microvessel density as a risk factor in squamous cell carcinoma of the penis. METHODS: Fifty patients with penile carcinoma were evaluated retrospectively. The mean age and standard deviation were 60.8±11.8 years. All of them were treated by penectomy and with positive nodes underwent groin lymphadenectomy. Tumor grading was 36 G1 and 24 G2/3. Primary lesion stage was 22 pT1 and 28 pT2-4. Positive inguinal nodes were observed in 18 patients. Selected paraffin embedded sections were submitted to CD34 immunohistochemical analysis by the avidin-biotin-immunoperoxidase method with antigen retrieval. All slides were examined using an automatic analyzer system and the number of micro-vessels in 10 high magnification power fields (400X) were counted in a blind analysis. RESULTS: Median number of microvessels was 631 in G1 versus 695 in G2/3 tumors (p=0.78), and 696 in pT1 versus 566 pT2-4 tumors (p=0.23). The respective data for pN0 patients was 525 and for pN+ was 696 (p=0.01), which is an unexpected result. CONCLUSION: CD34 immunoexpression or microvessel density determined by this method bear no association with tumor grade, stage or prognosis.

VEGF immunoexpression in penile carcinoma

OBJECTIVE: To investigate the vessel endothelial growth factor (VEGF) as a risk factor in squamous cell carcinoma of the penis (SCCP). METHODS: Forty-seven patients with penile carcinoma were evaluated retrospectively. The mean age and standard deviation were 61.1±11.7 years. All of them were treated by penectomy and those with positive nodes underwent groin lymphadenectomy. Tumor grading was 35 G1 and 12 G2/3. Primary lesion stage was 24 pT1 and 23 pT2-4. Positive inguinal nodes were observed in 15 patients. Selected paraffin embedded sections were submitted to VEGF immunohistochemical analysis by the avidin-biotin-immunoperoxidase method with antigen retrieval. All slides were examined using an automatic analyzer system and the proportion of labeled cells in 10 high magnification power fields (400X) were recorded in a blind analysis. RESULTS: Median (%) labeling index was 2.3 in G1 versus 2.2 in G2/3 tumors (p=0.60), and 4.0 in pT1 versus 1.8 pT2-4 tumors (p=0.10). The respective data for pN0 patients was 2.8 and for pN+ was 2.1 (p=0.20). Survival curves showed no association with patients survival. CONCLUSION: In squamous cell carcinoma of the penis the VEGF immunoexpression has no association with tumor grade or stage, as well as with patient survival.

Persistence of congenital penile frenulum in male cat: / Persistência de frênulo peniano congênito em gato macho:

The penile frenulum is a thin strip of connective tissue that connects the penis glans to the foreskin. Due to the few number of reports of this condition, this work reports the occurrence of persistent penile frenulum in a mixed breed domestic cat, of seven months old and 5.2 kg. In this particular case, penile frenulum was an incidental finding during clinical examination. The animal was in puberty, characterizing the penile frenulum. The resolution of the case occurred spontaneously prior to elective orchiectomy. Thus, the changes observed in the penis due to congenital alteration in cats are described in this report.  

O frênulo peniano é uma fina faixa de tecido conectivo que une a glânde ao prepúcio que pode se romper sob a ação da testosterona quando o animal inicia a puberdade. Devido a escassez de relatos desta afecção o presente trabalho relata a ocorrência de frênulo peniano persistente em um gato doméstico, sem raça definida, de sete meses de idade e 5,2 kg. O frênulo peniano foi um achado incidental durante o exame clínico do animal. O animal já apresentavase em puberdade, caracterizando o quadro de frênulo peniano. A resolução do caso ocorreu de maneira espontânea previamente a orquiectomia eletiva. Dessa forma, as alterações observadas no pênis devido a alteração congênita em gatos são descritas neste relato.  

VEGF immunoexpression in penile carcinoma

OBJECTIVE: To investigate the vessel endothelial growth factor (VEGF) as a risk factor in squamous cell carcinoma of the penis (SCCP). METHODS: Forty-seven patients with penile carcinoma were evaluated retrospectively. The mean age and standard deviation were 61.1±11.7 years. All of them were treated by penectomy and those with positive nodes underwent groin lymphadenectomy. Tumor grading was 35 G1 and 12 G2/3. Primary lesion stage was 24 pT1 and 23 pT2-4. Positive inguinal nodes were observed in 15 patients. Selected paraffin embedded sections were submitted to VEGF immunohistochemical analysis by the avidin-biotin-immunoperoxidase method with antigen retrieval. All slides were examined using an automatic analyzer system and the proportion of labeled cells in 10 high magnification power fields (400X) were recorded in a blind analysis. RESULTS: Median (%) labeling index was 2.3 in G1 versus 2.2 in G2/3 tumors (p=0.60), and 4.0 in pT1 versus 1.8 pT2-4 tumors (p=0.10). The respective data for pN0 patients was 2.8 and for pN+ was 2.1 (p=0.20). Survival curves showed no association with patients survival. CONCLUSION: In squamous cell carcinoma of the penis the VEGF immunoexpression has no association with tumor grade or stage, as well as with patient survival.

CD34 immunoexpression in penile carcinoma

OBJECTIVE: To investigate microvessel density as a risk factor in squamous cell carcinoma of the penis. METHODS: Fifty patients with penile carcinoma were evaluated retrospectively. The mean age and standard deviation were 60.8±11.8 years. All of them were treated by penectomy and with positive nodes underwent groin lymphadenectomy. Tumor grading was 36 G1 and 24 G2/3. Primary lesion stage was 22 pT1 and 28 pT2-4. Positive inguinal nodes were observed in 18 patients. Selected paraffin embedded sections were submitted to CD34 immunohistochemical analysis by the avidin-biotin-immunoperoxidase method with antigen retrieval. All slides were examined using an automatic analyzer system and the number of micro-vessels in 10 high magnification power fields (400X) were counted in a blind analysis. RESULTS: Median number of microvessels was 631 in G1 versus 695 in G2/3 tumors (p=0.78), and 696 in pT1 versus 566 pT2-4 tumors (p=0.23). The respective data for pN0 patients was 525 and for pN+ was 696 (p=0.01), which is an unexpected result. CONCLUSION: CD34 immunoexpression or microvessel density determined by this method bear no association with tumor grade, stage or prognosis.

AVALIAÇÃO DOS GÂNGLIOS REGIONAIS NO CARCINOMA EPIDERMÓIDE DO PÊNIS (CEP) ATRAVÉS DE UM ÍNDICE DE RISCO

The aim of this study was to investigate the predictive value of a risk index (RI) based on tumor pT stage and grade on the prognosis for inguinal lymphactic metastasis. We included 53 patients bearing schamous cell carcinoma of the penis (SCCP) treated by penectomy at the Hospital das Clínicas da FMRP-USP from 1978 to 1995. Age range varied from 32 to 97 years. Tumor grading was determined according to Broders method and staging by 1999 TNM system. RI was calculated by the sum of pT stage and grade (RI = pT+G). Bilateral inguinal lymphdenectomy was carried out within 6 weeks the penectomy in 14 patients cN+ at the presentation. The remaining 36 cases were kept under surveillance. In 9 of these patients required a delayed regional lymphadenectomy due to nodal relapse (median follow-up: 9 months). Proportions of pN0 patients according to RI were: 92% - 2, 80% - 3 and 0% - 6. The proportion of pN+ patients with a pooled RI of 4, 5 and 6 was significantly higher than that seen in pooled cases with lower RI (p = 0,004). From 36 patients initially cN0, 27 were RI 2 or 3 ( 5 of them progressed later on to pN+ , 19%) and 9 were RI ³ 4 (3 of them had progression to lymphatic spread, 33%). But, the difference observed in cN0 patients was not significant (p = 0,3). In conclusion, patients with RI ³ 4 were at higher risk of nodal spread when the overall sample was at stake, but this cutoff level for RI had no predictive value for the patients that were initially cN0.

O objetivo desse estudo foi investigar a validade de um índice de risco (IR) no prognóstico de metástases ganglionares no carcinoma epidermóide do pênis (CEP). Foram analisados 53 pacientes com CEP atendidos no Hospital das Clínicas da FMRP-USP, de janeiro de 1978 a dezembro de 1995. A idade variou de 32 a 97 anos. Os tumores foram graduados de acordo com a classificação de Broders e estadiados retrospectivamente (TNM 1999). O IR foi determinado pela soma de TeG (IR = T+G). A linfadenectomia inguinal bilateral (LD) foi indicada naqueles que apresentaram gânglios suspeitos após a antibioticoterapia ou que vieram apresentar alterações durante o período de observação. Faleceram 17 pacientes; A LD foi realizada em 14 pacientes e os 39 restantes permaneceram em observação clínica, sendo que em 8 deles foi feito LD após tempo mediano de 9 meses. Dos pacientes com índice de risco 2 e 3 92% e 80% respectivamente não apresentavam metastase ganglionar, ao passo que a totalidade daqueles com IR=6 tinha gânglios comprometidos. A associação dos grupos 2 e 3, comparada ao conjunto 4, 5 e 6, mostrou percentual maior de comprometimento nestes últimos (p = 0,0046). Dos 36 pacientes inicialmente No, 27 apresentavam IR 2 ou 3, dos quais 5 evoluíram para N+ (19%); os 9 restantes eram IR > 4 e 3 também evoluíram para N+ (33%). Apesar da diferença, não houve significância estatística entre eles (p = 0,3837). Embora tenha ocorrido maior comprometimento naqueles com IR > 4, o índice de risco não foi capaz de identificar previamente pacientes que evoluíram para N+.