Avaliação de diferentes fixadores na preservação das características histológicas de pele de orelha de cães / Evaluation of different fixatives for histological studies in canine ear skin

Os fixadores biológicos desempenham um papel importante na qualidade final da histologia. Na rotina veterinária, a biópsia de pele é um procedimento comum e a escolha do fixador é primordial para resultado final adequado. Os fixadores mais usados são à base de formalina, ainda que sejam tóxicos, cancerígenos, de baixa penetração e de fixação lenta. Mesmo assim, não existe um fixador ideal que substitua as suas qualidades. O objetivo deste trabalho foi avaliar qualitativamente a preservação das características histológicas de pele de cão utilizando diferentes fixadores de tecidos incluídos em parafina, cortados e corados pela hematoxilina-eosina. Utilizou-se uma caneta Punch de 4 milímetros para coletar amostras de pele de orelha em seis cadáveres de cães. Após coleta, os tecidos foram fixados em: (1) Bouin, durante seis horas; (2) Carnoy, durante quatro horas; (3) formaldeído tamponado 10% durante 24 horas, todos sob refrigeração (4ºC). Posteriormente, os tecidos foram processados, cortados e corados em hematoxilina e eosina. As lâminas foram avaliadas, às cegas, por quatro patologistas diferentes, que consideraram aspectos qualitativos a seguir: (1) qualidade da coloração; (2) preservação das características histológicas; e (3) preservação dos limites citoplasmáticos utilizando a escala de LIKERT de pontuação para cada lâmina. O fixador com a maior média de pontuação em todos os itens foi o formol tamponado com 3,76 pontos, seguido pelo Bouin (3,39) e pelo Carnoy (2,52). O formol pode trazer riscos à saúde do profissional que rotineiramente o manuseia, portanto se faz necessária a busca por fixadores com as mesmas qualidades, mas menos nocivos à saúde.(AU)

The biological fixatives have an important role in the final histology quality. In veterinary, routine skin biopsy is a common procedure and the choice of fixative is essential for the final result. The most common fixative is Formalin, even though it is toxic, carcinogenic, and has low and slow penetration. Still, there isn't a fixer which can replace the qualities of formalin. The aim of this study was to evaluate qualitatively the preservation of the histological features of dog skin using different tissue fixative embedded in paraffin, sectioned and stained with hematoxylin - eosin. We used a 4 mm punch pen to collect ear skin samples in six dog cadavers. After collection, the tissues were fixed in: (1) Bouin for 6 hours; (2) Carnoy for 4 hours; (3) 10% buffered formaldehyde for 24 hours, all under refrigeration (4 ° C). The tissues were then processed, sectioned and stained with hematoxylin and eosin. The slides were evaluated blindly by four different pathologists who considered the qualitative aspects below: (1) quality of coloring; (2) preservation of the histological characteristics; (3) preservation of cytoplasmic boundaries using a Likert scale score for each blade. The fixative with the highest mean score on all items was buffered formalin with 3.76 points followed by Bouin (3.39) and Carnoy (2.52). Formaldehyde can bring health a risk of professional routine handling, so it is necessary to search for a biological fixative with the same qualities being less harmful to health.(AU)

Avaliação de diferentes fixadores na preservação das características histológicas de pele de orelha de cães / Evaluation of different fixatives for histological studies in canine ear skin

Os fixadores biológicos desempenham um papel importante na qualidade final da histologia. Na rotina veterinária, a biópsia de pele é um procedimento comum e a escolha do fixador é primordial para resultado final adequado. Os fixadores mais usados são à base de formalina, ainda que sejam tóxicos, cancerígenos, de baixa penetração e de fixação lenta. Mesmo assim, não existe um fixador ideal que substitua as suas qualidades. O objetivo deste trabalho foi avaliar qualitativamente a preservação das características histológicas de pele de cão utilizando diferentes fixadores de tecidos incluídos em parafina, cortados e corados pela hematoxilina-eosina. Utilizou-se uma caneta Punch de 4 milímetros para coletar amostras de pele de orelha em seis cadáveres de cães. Após coleta, os tecidos foram fixados em: (1) Bouin, durante seis horas; (2) Carnoy, durante quatro horas; (3) formaldeído tamponado 10% durante 24 horas, todos sob refrigeração (4ºC). Posteriormente, os tecidos foram processados, cortados e corados em hematoxilina e eosina. As lâminas foram avaliadas, às cegas, por quatro patologistas diferentes, que consideraram aspectos qualitativos a seguir: (1) qualidade da coloração; (2) preservação das características histológicas; e (3) preservação dos limites citoplasmáticos utilizando a escala de LIKERT de pontuação para cada lâmina. O fixador com a maior média de pontuação em todos os itens foi o formol tamponado com 3,76 pontos, seguido pelo Bouin (3,39) e pelo Carnoy (2,52). O formol pode trazer riscos à saúde do profissional que rotineiramente o manuseia, portanto se faz necessária a busca por fixadores com as mesmas qualidades, mas menos nocivos à saúde.(AU)

The biological fixatives have an important role in the final histology quality. In veterinary, routine skin biopsy is a common procedure and the choice of fixative is essential for the final result. The most common fixative is Formalin, even though it is toxic, carcinogenic, and has low and slow penetration. Still, there isn't a fixer which can replace the qualities of formalin. The aim of this study was to evaluate qualitatively the preservation of the histological features of dog skin using different tissue fixative embedded in paraffin, sectioned and stained with hematoxylin - eosin. We used a 4 mm punch pen to collect ear skin samples in six dog cadavers. After collection, the tissues were fixed in: (1) Bouin for 6 hours; (2) Carnoy for 4 hours; (3) 10% buffered formaldehyde for 24 hours, all under refrigeration (4 ° C). The tissues were then processed, sectioned and stained with hematoxylin and eosin. The slides were evaluated blindly by four different pathologists who considered the qualitative aspects below: (1) quality of coloring; (2) preservation of the histological characteristics; (3) preservation of cytoplasmic boundaries using a Likert scale score for each blade. The fixative with the highest mean score on all items was buffered formalin with 3.76 points followed by Bouin (3.39) and Carnoy (2.52). Formaldehyde can bring health a risk of professional routine handling, so it is necessary to search for a biological fixative with the same qualities being less harmful to health.(AU)

Microtia e atresia congênita do canal auditivo em cão: relato de caso / Microtia and congenital external auditory canal atresia in dog: case report

Descreve-se o caso de um cão, Pinscher, fêmea de 11 meses de idade, com alteração anatômica da orelha do lado direito. Após avaliação, foi realizado o diagnóstico clínico e o radiográfico de microtia com atresia do canal auditivo do lado direito. Como o animal não apresentava sinais de alterações no sistema vestibular ou otite, optou-se pelo acompanhamento clínico do caso. A microtia, caracterizada pela hipoplasia parcial ou completa da pina, assim como outras anomalias do conduto auditivo, é raramente descrita em medicina veterinária. De acordo com a revisão de literatura realizada, este é o primeiro relato ocorrido no Brasil de um cão com microtia e atresia congênita do canal auditivo sem a associação de síndrome vestibular.(AU)

This paper describes an 11-month-old female Pinscher dog, with anatomical changes of the ear on the right side. After physical exam, clinical and radiographic diagnosis of microtia with ear canal atresia on the right side was made. As the animal did not show any signs of vestibular system changes or ear infections, the veterinarians opted for monitoring the patient. Microtia, characterized by the partial or complete hypoplasia of pina, as well as other abnormalities of ear canal are rarely described in veterinary medicine. As far as the author´s knowledge by literature review carried out, this is the first report of a dog with microtia and congenital ear canal atresia not associated with vestibular syndrome in Brazil.(AU)

Microtia e atresia congênita do canal auditivo em cão: relato de caso / Microtia and congenital external auditory canal atresia in dog: case report

Descreve-se o caso de um cão, Pinscher, fêmea de 11 meses de idade, com alteração anatômica da orelha do lado direito. Após avaliação, foi realizado o diagnóstico clínico e o radiográfico de microtia com atresia do canal auditivo do lado direito. Como o animal não apresentava sinais de alterações no sistema vestibular ou otite, optou-se pelo acompanhamento clínico do caso. A microtia, caracterizada pela hipoplasia parcial ou completa da pina, assim como outras anomalias do conduto auditivo, é raramente descrita em medicina veterinária. De acordo com a revisão de literatura realizada, este é o primeiro relato ocorrido no Brasil de um cão com microtia e atresia congênita do canal auditivo sem a associação de síndrome vestibular.(AU)

This paper describes an 11-month-old female Pinscher dog, with anatomical changes of the ear on the right side. After physical exam, clinical and radiographic diagnosis of microtia with ear canal atresia on the right side was made. As the animal did not show any signs of vestibular system changes or ear infections, the veterinarians opted for monitoring the patient. Microtia, characterized by the partial or complete hypoplasia of pina, as well as other abnormalities of ear canal are rarely described in veterinary medicine. As far as the author´s knowledge by literature review carried out, this is the first report of a dog with microtia and congenital ear canal atresia not associated with vestibular syndrome in Brazil.(AU)

Anatomical study of the pigs temporal bone by microdissection

PURPOSE: Initial study of the pig`s temporal bone anatomy in order to enable a new experimental model in ear surgery. METHODS: Dissection of five temporal bones of Sus scrofa pigs obtained from UNIFESP - Surgical Skills Laboratory, removed with hole saw to avoid any injury and stored in formaldehyde 10% for better conservation. The microdissection in all five temporal bone had the following steps: inspection of the outer part, external canal and tympanic membrane microscopy, mastoidectomy, removal of external ear canal and tympanic membrane, inspection of ossicular chain and middle ear. RESULTS: Anatomically it is located at the same position than in humans. Some landmarks usually found in humans are missing. The tympanic membrane of the pig showed to be very similar to the human, separating the external and the middle ear. The middle ear`s appearance is very similar than in humans. The ossicular chain is almost exactly the same, as well as the facial nerve, showing the same relationship with the lateral semicircular canal. CONCLUSION: The temporal bone of the pigs can be used as an alternative for training in ear surgery, especially due the facility to find it and its similarity with temporal bone of the humans. .(AU)

Micro-endoscopic ear anatomy of guinea pig applied to experimental surgery

PURPOSE: To describe topographic and endoscopic anatomy of guinea pig ear for development of surgical approaches in experimental studies. METHODS: Experimental study. Eight adult guinea pigs (Cavia porcellus) were used in this study. Four animals were described through endoscopic view and four animals were used to describe topographic anatomy. RESULTS: The main structures of middle ear were well identified through endoscopy view: oval and round window, ossicles and vascular structures. Temporal bone position, landmarks and its relations to skull are perceived with topographic description. CONCLUSION: Topographic anatomic description allowed exposition of temporal bone relations for external surgical approaches. Alternatively, grooves and middle ear structures were identified and may be used to transcanal accesses. .(AU)

Tissue response evaluation of the mucosa of the tympanic cavity of guinea pigs, when receiving biodegradable implant

PURPOSE: To evaluate the tissue response of the mucosa of the tympanic cavity of guinea pigs, when receiving biodegradable implant. METHODS: A total of 20 male guinea pigs were divided into 2 groups. After paracentesis in both ears, a biodegradable polymer of poly lactic-co-glycolic acid was implanted in only one middle ear. Histological analysis using neutrophil exudate and vascular neoformation (acute inflammation) and fibroblast proliferation and mononuclear inflammatory cells (chronic inflammation) as parameters was performed after 10 and 30 days of survival (groups 1 and 2, respectively). RESULTS: Four ears in group 1 and 7 in group 2 had an increase of neutrophil exudate. Vascular neoformation occurred in ears with or without the implant, in both groups. Fibroblast proliferation and mononuclear inflammatory cells (lymphocytes and macrophages) increased in ears with implant in group 2. CONCLUSION: The tissue response by histological analysis of the mucosa of the tympanic cavity of guinea pigs, when receiving biodegradable implant, showed no statistically significant difference between ears with or without the implant. .(AU)

Diagnóstico da otite média em cães: comparação entre radiografia convencional, canalografia e audiometria de impedância / Diagnosis of otitis media in dogs: comparison between conventional radiography, canalography and impedance audiometry

Radiography (conventional and contrasted - canalography) and audioelectrophysiologic (impedance audiometry) tests were correlated with otitis media in 12 middle ear affected and 12 healthy dogs. When comparing both diagnostic methods, it was found that the reliability in the detection of otitis media in dogs was 100 percent for the first and 58.3-60.0 percent for the latter. Thus, it was possible to attest that the impedance audiometry was the most accurate method in the identification of otitis media in dogs when compared with radiographic assessments.(AU)

Effect of the period of extrinsic mechanical compression following sclerotherapy in veins in rabbit ears / Efeito do tempo da compressão mecânica extrínseca após escleroterapia em veias de orelhas de coelhos

PURPOSE: Research whether a post-sclerotherapy venous compression period of up to 120 hours is sufficient to avoid reperfusion in treated veins; whether there is a relationship between the inflammatory intensity in venous walls and adjacent tissue and the size of venous thrombosis; whether the intensity of the post-sclerotherapy inflammation varies with the period of compression; whether there is a relationship between the presence of hemosiderin in the tissues adjacent to the sclerosing blood vessels and venous blood clots. METHODS: Twenty eight rabbits, all male, were utilized, distributed into four groups (0, 24, 72 and 120). All the animals were administered with 0.25 ml of 1% polidoconal solution and, as a control, 0.25 ml of 0.9% sodium chloride solution in the marginal dorsal vein of the right and left ears, respectively. Mechanical compression was applied to the perfused stretch of the vein, except for the animals in group 0. The period of compression varied from 0 to 120 hours in the groups. An anatomopathological examination of the section of the right and left marginal dorsal veins of all the animals was conducted. RESULTS: There was no significant difference among the various compression periods, both in terms of the degree of vein thrombosis and in the inflammatory intensity in both ears of the various groups. A positive and significant correlation was observed between the inflammatory intensity and the size of the thrombus and in the occurrence of thrombi and hemosiderin. CONCLUSIONS: A compression period of up to 120 hours is not sufficient to prevent reperfusion in sclerosing blood vessels. The intensity of tissue inflammation is related to the size of the thrombus, but not to the compression period. The presence of hemosiderin in the tissues adjacent to the vessels subjected to sclerosis is related to the presence of venous coagulation.(AU)

OBJETIVO: Pesquisar se o tempo de compressão venosa de até 120 horas pós-escleroterapia é suficiente para evitar reperfusão nas veias tratadas; se há relação entre a intensidade inflamatória na parede venosa e tecidos adjacentes e o tamanho do trombo venoso; se a intensidade da inflamação pós-escleroterapia varia com o tempo de compressão; se há relação entre a presença de hemossiderina nos tecidos adjacentes ao vaso esclerosado e coágulo venoso. MÉTODOS: Utilizaram-se 28 coelhos, machos, distribuídos em quatro grupos (0, 24, 72 e 120). Em todos os animais foram administrados 0,25 ml de solução de polidocanol 1% e, como controle, 0,25 ml de solução de cloreto de sódio 0,9% na veia marginal dorsal das orelhas direita e esquerda, respectivamente. Realizou-se compressão mecânica em trecho da veia perfundida, exceto nos animais do grupo 0. O tempo de compressão variou de 0 a 120 horas nos grupos. Realizou-se exame anatomopatológico de trecho das veias marginais dorsais direita e esquerda de todos os animais. RESULTADOS: Não houve diferenças significativas nos diversos tempos de compressão, tanto no grau de trombose venosa como na intensidade inflamatória, em ambas as orelhas, nos diversos grupos. Observou-se correlação positiva e significativa entre intensidade inflamatória e tamanho do trombo e na ocorrência de trombos e hemossiderina. CONCLUSÕES: O tempo de até 120 horas de compressão não é suficiente para evitar reperfusão nos vasos esclerosados. A intensidade da inflamação nos tecidos tem relação com o tamanho do trombo, mas não com o tempo de compressão. A presença de hemossiderina nos tecidos adjacentes ao vaso submetido à esclerose está relacionada com a presença de coágulo venoso. DESCRITORES: Escleroterapia. Soluções Esclerosantes. Varizes. Experimentação Animal. Dissertações Acadêmicas. Coelhos.(AU)

Inflammatory response, parasite load and AgNOR expression in ear skin of symptomatic and asymptomatic Leishmania (Leishmania) chagasi infected dogs

The skin has an important role in the transmission of visceral leishmaniasis (VL) as the infection pathway in dogs. To better characterize the inflammatory response of intact skin in VL, sixty infected dogs (30 symptomatic and 30 asymptomatic) and six non-infected controls were studied. Diagnosis of visceral leishmaniasis was confirmed by RIFI and ELISA; direct visualization of the parasite in bone marrow aspirate; imprints of popliteal lymph nodes, spleen, liver and skin; culture in NNN-phase liquid Schneider's medium; and PCR (performed only in the ear skin). Amastigote forms of the parasite in intact skin were found only in symptomatic dogs. Inflammatory infiltrates were observed in all groups, varying from intense and/or moderate in symptomatic to discrete and/or negligible in asymptomatic and control animals. Parasite load was associated with the intensity of the inflammatory response and with clinical manifestations in canine visceral leishmaniasis. AgNOr as active transcription markers were expressed in inflammatory cells and within apoptotic bodies in all groups, including controls, with no statistical difference. Therefore, cell activation and transcription do occur in both symptomatic and asymptomatic canine visceral leishmaniasis and may result in more necrosis and inflammation or in apoptosis and less symptoms, depending on the parasite load.(AU)

Efficacy of Argentine propolis formulation for topical treatment of canine otitis externa / Eficácia da formulação da propólis Argentina para o tratamento tópico de otite externa canina

The therapeutic effects of Argentine propolis ear drop formulation on canine otitis externa were evaluated. Forty-eight dogs with symptoms of otitis externa were randomly assigned to double-blinded, controlled clinical trial to evaluate the efficacy of topical formulation with propolis versus a topical placebo in the treatment of otitis externa. The propolis preparation and placebo were administrated into both external ear canals, twice daily for 14 days. Throughout the study, clinical examination and microbiological analysis of dogs ear exudates were made. The most frequent microorganisms isolated in culture media were: Malassezia pachydermatis (54.2 percent), Staphylococcus aureus (43.8 percent), coagulase-negative Staphylococcus (25.0 percent), Pseudomonas aeruginosa (20.8 percent), Candida albicans (18.8 percent), Proteus mirabilis (16.7 percent), Streptococcus spp. (16.7 percent), Enteroccocus faecalis (12.5 percent), Escherichia coli (12.5 percent), Staphylococcus intermedius (6.3 percent), Klebsiella spp. (4.2 percent), andCandida glabrata (2.1 percent). Whereas the control group did not recover from the infectious ear disease, the propolis preparation exhibited antimicrobial activity against most of the microorganisms isolated from samples of the treated group. In addition, no propolis-adverse effects were observed. This allowed propolis-treated patients to show a significant improvement of the clinical parameters. Thus, this new Argentine propolis ear drop formulation may be used for topical treatment of otitis externa in dogs.(AU)

Os efeitos terapêuticos da formulação em gotas óticas de própolis procedentes da Argentina foram avaliados no tratamento da otite externa canina. Quarenta e oito cães com sintomas de otite externa foram distribuídos aleatoriamente em ensaio clínico duplo-cego controlado para avaliar a eficácia da formulação tópica com a própolis contra um placebo tópico no tratamento da otite externa. A preparação de própolis e placebo foi administrada em ambos os canais da orelha externa, duas vezes por dia, durante 14 dias. Ao longo do estudo, os cães foram submetidos a exame físico e à análise microbiológica de exsudatos auriculares. Os mais frequentes microrganismos isolados em meios de cultura foram: Malassezia pachydermatis (54,2 por cento), Staphylococcus aureus (43,8 por cento), Staphylococcus coagulase-negativo (25,0 por cento), Pseudomonas aeruginosa (20,8 por cento), Candida albicans (18,8 por cento), Proteus mirabilis (16,7 por cento), Streptococcus spp.(16,7 por cento), Enterococcus faecalis (12,5 por cento), Escherichia coli (12,5 por cento), Staphylococcus intermedius (6,3 por cento), Klebsiella spp.(4,2 por cento) e Candida glabrata (2,1 por cento). A preparação de própolis apresentou atividade antimicrobiana contra a maioria dos microrganismos isolados de amostras do grupo de tratamento, sendo que os do grupo-controle não se recuperaram da doença infecciosa auricular, e não foram observados efeitos adversos à própolis. Isso permitiu aos pacientes tratados com própolis melhora significativa dos parâmetros clínicos. Essa nova formulação da própolis argentina para o ouvido apresenta potencial utilidade no tratamento tópico da otite externa em cães.(AU)