RESUMO
This study aimed to evaluate the ovicidal activity of the hydroalcoholic extract of Schinus terebinthifolia (SCH; T1) against Ancylostoma spp. and its influence of storage time in the extract stored for 36 months (T36). Eggs of Ancylostoma spp. were obtained from naturally parasitized dogs, and used for the larval hatchability test, where the eggs were exposed to T1 and T36 extracts of SCH (15-0.625 mg/mL). In T1, all concentrations inhibited more than 80% of the eggs, being 100% at concentrations between 15 and 5 mg/mL (p > 0.05). At T36, all concentrations were active, even the ones between 2.5 and 0.625 mg/mL, with 100% inhibition (p < 0.05), revealing that the storage time maintained the ovicidal action. By LC-MS, T36 presented ethyl gallate, myricitrin, and gallic acid as major compounds. These findings support the promising use of SCH extract as an ovicide against Ancylostoma spp., even stored for 36 months of shelf life.
Assuntos
Anacardiaceae , Cães , Animais , Ancylostoma , Extratos Vegetais/farmacologia , Larva , Espectrometria de MassasRESUMO
We evaluated the chemical composition, toxicity, and antibacterial activity of Schinus terebinthifolia (SCH), Eugenia uniflora (EUG), Persicaria hydropiperoides (PER), Equisetum hyemale (EQU), Solidago chilensis (SOL), and Baccharis trimera (BRA). These plants were tested (7.5-0.01 mg/mL) against Gram-positive (G+; n = 32) and Gram-negative (G-; n = 26) isolates from animals (M07-A9, CLSI). Antibiogram (disk diffusion), chromatographic analysis (UPLC), and toxicity assay (HET-CAM) were also performed. A high incidence of resistance was noted, in which 18.4% (07/38) of G+ (Staphylococcus intermedius/Enterococcus faecium) and 17.7% (06/34) of G- (Pseudomonas aeruginosa/Escherichia coli/Proteus mirabilis) were multidrug-resistant. All bacteria were sensitive (MIC50) to SCH (both 3.75 mg/mL), EUG (3.75 mg/mL and 7.5 mg/mL, respectively) and PER (both 7.5 mg/mL). SCH/EUG/PER highlighted as antibacterial, probably due to the major compounds (ethyl gallate, quinic acid, quercetin). These extracts showed normal embryonic development (SCH/EUG: 7.5-0.94 mg/mL). These findings highlighted the promising use of native plants for therapeutic purposes.
Assuntos
Animais Selvagens , Bactérias , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Brasil , Escherichia coli , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Testes de Sensibilidade MicrobianaRESUMO
We aimed to evaluate the chemical, antioxidant, cytotoxic, and antifungal activities of hydroalcoholic extracts of native plants from Southern Brazil: Schinus terebinthifolia (SCH), Persicaria hydropiperoides (PER), Eugenia uniflora (EUG) and Equisetum hyemale (EQU). Ethyl gallate, quercetin, and quinic acid were prevalent compounds identified by LC-MS. For total phenolic/flavonoid contents and the antioxidant potential against ABTS/DPPH radicals, the ascending order was EQU < PER < EUG < SCH. All extracts were low cytotoxic and kept a high Vero cell viability (>75%) at concentrations up to 12.5 mg/mL (MTT assay). By M38-A2/M27-A3 (CLSI) against 68 clinical isolates of animals and strains of Malassezia pachydermatis, Sporothrix brasiliensis, Microsporum canis, Microsporum gypseum and Trichophyton mentagrophytes, all extracts (MIC/MFC ≤3.13-100 mg/mL) were active, except EUG. SCH inhibited and killed S. brasiliensis (MIC/MFC50/90 3.12-12.5 mg/mL) and dermatophytes (MIC/MFC 6.25-25 mg/mL) resistant to ketoconazole and itraconazole. These findings support the promising use of the selected plant extracts as antifungal agents.
Assuntos
Doenças do Gato , Doenças do Cão , Animais , Antifúngicos/química , Antioxidantes/farmacologia , Brasil , Gatos , Cães , Testes de Sensibilidade Microbiana , Extratos Vegetais/químicaRESUMO
Thirty Wistar rats subcutaneously infected by an itraconazole-resistant Sporothrix brasiliensis received the oral daily treatment (n = 10, each) of control (CTL, saline solution), itraconazole (ITZ, 10 mg/kg) and marjoram essential oil (MRJ, 80 mg/kg) for 30 days. Weekly, the clinical evaluation and euthanasia for histopathology and fungal burden were performed. Only animals from MRJ evolved to the remission of the cutaneous lesion with a mild to absent presence of yeasts in footpad, besides decreased the fungal burden in the systemic organs compared to CTL and ITZ (p < 0.05), preventing the fungal spread, mainly in the liver and spleen. The antifungal activity may have been attributed to the majority composition of terpinen-4-ol (34.09%), γ-terpinene (14.28%) and α-terpinene (9.6%), which the mode of action was at the level of ergosterol complexation. These findings highlighted the antifungal and the systemic protective effects of MRJ, supporting the promising use in the treatment of cutaneous sporotrichosis.
Assuntos
Antifúngicos/farmacologia , Óleos Voláteis , Origanum , Sporothrix , Esporotricose , Animais , Antifúngicos/isolamento & purificação , Testes de Sensibilidade Microbiana , Óleos Voláteis/farmacologia , Origanum/química , Compostos Fitoquímicos/isolamento & purificação , Compostos Fitoquímicos/farmacologia , Ratos , Ratos Wistar , Sporothrix/efeitos dos fármacos , Esporotricose/tratamento farmacológicoRESUMO
Osteosarcoma is characterized by the production of osteoid or woven bone, using neoplastic osteoblasts. It is the most common primary bone neoplasm in canines and humans. This neoplasm was previously reported in all vertebrate classes, including a wide variety of mammals. However, there is no case report describing this neoplasm in Didelphis albiventris. Therefore, the objective of this manuscript is to describe the clinical-pathological aspects of fibroblastic osteosarcoma in D. albiventris. A wild adult male white-eared opossum (D. albiventris) arrived at the zoological park of the "Universidade de Caxias do Sul" with a swollen left thoracic limb. After a general clinical examination, the animal was transferred to the Veterinary Clinic of "Universidade de Caxias do Sul" for radiographic examination of the left thoracic limb and thorax. Additionally, some material was collected through fine needle aspiration (FNA) for cytologic evaluation. The radiographic findings and cytologic evaluation indicated osteosarcoma. The animal was euthanized due to severe clinical conditions and guarded prognosis. During necropsy, macroscopic analysis of the viscera was performed, fragments of various organs were collected and fixed in 10% neutral buffered formalin. All fragments were processed following routine histological techniques. The histopathological evaluation confirmed osteosarcoma, which was classified as a fibroblastic subtype. Case reports are crucial for the knowledge of incidence, prevalence, and behavior of the current mentioned disease, as well as other diseases, in species with such limited information. In order to obtain a decisive diagnosis, a few different examination methods were associated. Although the observations presented are based on a single case, this neoplasm had a similar clinical presentation to that described in other species.(AU)
O osteossarcoma é caracterizado pela produção de osteoide ou osso imaturo, por osteoblastos neoplásicos. É a neoplasia óssea primária mais comum em caninos e humanos. Essa neoplasia já foi relatada em todas as classes de vertebrados, incluindo uma grande variedade de mamíferos. Não havendo descrição dessa neoplasia até o momento em Didelphis albiventris. O objetivo deste trabalho é descrever aspectos clínico-patológicos de um caso de osteossarcoma fibroblástico em D. albiventris. Chegou para atendimento no Zoológico da Universidade de Caxias do Sul um gambá-de-orelha-branca (D. albiventris), macho, adulto, de vida livre com aumento de volume no membro torácico esquerdo. Após avaliação clínica geral, o animal foi encaminhado para a Clínica Veterinária da Universidade de Caxias do Sul para realização de radiografia do membro torácico esquerdo e de tórax, sendo também realizada coleta de material por punção aspirativa por agulha fina (PAAF) para avaliação citológica. Os achados radiográficos e da avaliação citológica foram sugestivos de osteossarcoma. Devido ao estado clínico grave e prognóstico reservado optou-se pela eutanásia. Durante a necropsia realizou-se a análise macroscópica das vísceras, foram coletados fragmentos de diversos órgãos, fixados em formalina 10%, processados pelas técnicas histológicas de rotina. Na avaliação histopatológica confirmou-se a suspeita de osteossarcoma sendo classificado no subtipo fibroblástico. A descrição de relatos é fundamental para conhecimento da incidência, prevalência e comportamento desta e de outras doenças em espécies que as informações são limitadas. A associação de diferentes métodos de exames foram necessários para a obtenção de um diagnóstico definitivo. Embora as observações apresentadas se baseiem em um caso único, esta neoplasia possuiu apresentação clínica semelhante a descrita em outras espécies.(AU)
Assuntos
Animais , Neoplasias Ósseas/veterinária , Osteossarcoma/diagnóstico , Osteossarcoma/veterinária , Osteossarcoma/diagnóstico por imagem , Didelphis , Radiografia Torácica/veterináriaRESUMO
In Brazil, a significant percentage of the population lives without basic sanitation, experiencing so-called social vulnerability. The fact that these people share the environment with animals promotes the establishment of zoonotic parasitic infections, as well as the resultant parasitic cycles. Thus, parasites present in the environment must be identified, so that control measures can be recommended. In this context, this study's objective was to evaluate environmental contamination by parasitic forms in a socially vulnerable community in southern Rio Grande do Sul. A total of 100 soil samples collected from the community were processed by a sodium dichromate centrifuge-flotation technique and analyzed by a compound microscope (40X objective) for the identification of parasite eggs, oocysts and cysts. All points were positive for two or more parasites, with the identification of 33.59% non-identified coccidian oocysts, Strongylida (25.4%), Ascaridida (21.31%), Trichuris spp. (8.19%), Toxocara spp. (3.27%), Amoebas (4.08%), Dioctophyma renale (2.45%), and Giardia spp.(1.63%). The presence of parasitic forms in all points analyzed surpasses other studies of environmental contamination carried out in the southern region of Brazil. In addition, the identification of several parasitic forms with zoonotic potential is concerning, since it shows the possibility of parasitic transmission to humans and other animals. In view of the results, the conclusion is that the environment analyzed is contaminated by parasitic forms, constituting a serious public health problem. Therefore, implementing educational and preventive measures in the community to control parasites is of crucial importance.(AU)
No Brasil, uma parcela significativa da população não possui saneamento básico e vive em situação de vulnerabilidade social, compartilhando o ambiente com animais, possibilitando o estabelecimento de infecções parasitárias zoonóticas e a manutenção do ciclo dos parasitos. Assim, para que medidas de controle sejam preconizadas, torna-se necessário a identificação dos parasitos presentes no ambiente. Neste contexto, este trabalho avaliou a contaminação ambiental por formas parasitárias em comunidade de vulnerabilidade social no sul do Rio Grande do Sul. Foram coletadas cem amostras de solo da comunidade, que foram processadas pela técnica de centrifugo-flutuação em solução de dicromato de sódio e analisadas em microscópio composto (objetiva 40X) para a identificação dos ovos, oocistos e cistos de parasitos. Todos os pontos de coleta foram positivos para dois ou mais parasitos, sendo diagnosticados oocistos de coccídios não-identificados (33,59%), Strongylida (25,4%), Ascaridida (21,31%), Trichuris spp. (8,19%), Toxocara spp. (3,27%), Amebas (4,08%), Dioctophyma renale (2,45%), Giardia spp. (1,63%). A quantidade de formas parasitárias em todos os pontos analisados supera a contida em outros estudos de contaminação ambiental já realizados na região sul do Brasil. Além disso, a identificação de diversas formas parasitárias com potencial zoonótico é preocupante, pois evidencia a possibilidade de transmissão de parasitoses ao homem e a outros animais. Diante dos resultados, conclui-se que o ambiente em questão está contaminado por formas parasitárias, constituindo um sério problema de saúde pública. Ressalta-se a importância da implantação de medidas educativas e preventivas com a comunidade para o controle dos parasitos.(AU)
Assuntos
Humanos , Animais , Doenças Parasitárias em Animais/epidemiologia , Doenças Parasitárias/epidemiologia , Vulnerabilidade Social , Solo/parasitologia , Zoonoses/etiologia , Saneamento Básico , Brasil/epidemiologiaRESUMO
As aflatoxinas pertencem ao grupo das micotoxinas e são substâncias sintetizadas durante a multiplicação de fungos que podem contaminar alimentos e rações animais, produzindo efeitos agudos ou crônicos naqueles que os consomem. Os cães são sensíveis aos efeitos tóxicos das aflatoxinas, havendo dificuldades de diagnóstico definitivo desta doença devido a fatores, como a inespecificidade dos sinais clínicos e o custo elevado da análise da presença de toxinas na alimentação. Desta forma, o objetivo deste artigo foi relatar um surto de aflatoxicose em dezoito caninos, no município de Pelotas - RS, no qual apenas um canino não veio a óbito, após a ingestão de quirelas de milho e arroz contaminadas. O diagnóstico definitivo de aflatoxicose aguda foi obtido através dos dados da anamnese, exames complementares, resultados de necropsia e histopatologia, além de análise da presença de toxinas na alimentação dos animais. Na necropsia observou-se degeneração e necrose hepática severa e na análise das amostras de alimento foi constatada a presença de aflatoxinas B1, B2, G1 e G2, em um somatório de 898,2μg/kg na quirela de milho e de 409,86μg/kg na de arroz. Assim sendo, medidas de conscientização acerca do controle e prevenção das micotoxicoses devem ser tomadas, salientando os possíveis danos à saúde dos animais. Além disto, métodos de identificação de micotoxinas, devem ser implementados na rotina hospitalar, incluindo a análise da ração oferecida, visto que a aflatoxicose, muitas vezes é subestimada.(AU)
Aflatoxins are mycotoxins synthesized during fungal multiplication and are able to contaminate food and animal feed, producing acute or chronic effects on consumers. Dogs are sensitive to this substance toxic. Disease final diagnosis has been hard to be made due to several factors, such as lack of specific clinical signs and high costs of testing for toxin presence in food. Thus, this paper aimed to report an aflatoxicosis outbreak in eighteen dogs in Pelotas - RS, Brazil. Only one dog has not come to death, after eating contaminated crushed corn and rice. Final diagnosis of acute aflatoxicosis was established through anamnesis, laboratory tests, necropsy and histopathological results, besides analysis of toxin presence in animal feed. At necropsy, degeneration and severe hepatic necrosis were observed and the analysis of food samples confirmed B1, B2, G1 and G2 aflatoxins, in an amount of 898.2μg/kg in crushed corn and 409.86μg/kg in rice. Therefore, awareness measures about control and prevention of mycotoxicosis should be taken, stressing the possible damage to the health of animals. Furthermore, mycotoxin identification methods must be implemented into routine clinical practice, including analysis of animal feed offered, since aflatoxicosis has often been underestimated.(AU)
Assuntos
Animais , Cães , Aflatoxinas/intoxicação , Aflatoxinas/toxicidade , Ração Animal/microbiologia , Grão Comestível/microbiologiaRESUMO
Abstract Dioctophyme renale is a zoonotic parasite with worldwide distribution, although its occurrence is little known. The objective here was to evaluate the presence of parasite eggs in the environment and in the urine of dogs and cats in an urban area. Soil samples and urine were evaluated respectively by means of the Caldwell-Caldwell technique and urinalysis. Out of the 100 soil samples, 3% presented D. renale eggs, and out of the 43 urine samples, 18.6% were positive, including the feline samples. Thus, D. renale eggs are present in the urban environment, and dogs and cats are parasitized by this nematode, which therefore represents a risk to public health.
Resumo Dioctophyme renale é um parasito zoonótico com distribuição cosmopolita, entretanto sua ocorrência é pouco conhecida. Assim, o objetivo foi avaliar a presença de ovos do parasito no ambiente e na urina de cães e gatos de zona urbana. As amostras de solo e urina foram avaliadas, respectivamente, pela técnica de Caldwell e Caldwell e por urinálise. Das 100 amostras de solo, 3% apresentaram ovos de D. renale, e das 43 amostras de urina, 18,6% foram positivas, incluindo a de felino. Conclui-se que ovos de D. renale estão presentes no ambiente urbano, assim como os caninos e felinos apresentam-se parasitados, representando um risco para a saúde pública.
Assuntos
Animais , Gatos , Cães , Solo/parasitologia , Urina/parasitologia , Dioctophymatoidea/isolamento & purificação , Contagem de Ovos de Parasitas/veterináriaRESUMO
Dioctophyme renale is a zoonotic parasite with worldwide distribution, although its occurrence is little known. The objective here was to evaluate the presence of parasite eggs in the environment and in the urine of dogs and cats in an urban area. Soil samples and urine were evaluated respectively by means of the Caldwell-Caldwell technique and urinalysis. Out of the 100 soil samples, 3% presented D. renale eggs, and out of the 43 urine samples, 18.6% were positive, including the feline samples. Thus, D. renale eggs are present in the urban environment, and dogs and cats are parasitized by this nematode, which therefore represents a risk to public health.
Assuntos
Dioctophymatoidea/isolamento & purificação , Solo/parasitologia , Urina/parasitologia , Animais , Gatos , Cães , Contagem de Ovos de Parasitas/veterináriaRESUMO
O objetivo deste trabalho foi identificar plantas utilizadas como medicinais e aquelas com potencial tóxico localizadas no bairro Simões Lopes, Padre Réus e Porto em Pelotas-RS. Durante um ano foram entrevistados 111 proprietários de animais atendidos no Ambulatório Veterinário-Universidade Federal de Pelotas, com o propósito de avaliar o conhecimento, cultivo, utilização e forma de obtenção das plantas medicinais, faixa etária e o sexo das pessoas, número de animais nas residências e resultados do uso de plantas nos animais. Após investigação inicial, realizaram-se visitas na comunidade com intuito de fotografar e coletar amostras das plantas para identificação botânica. Foram listadas 74 plantas com fins medicinais, principalmente das famílias Asteraceae e Lamiaceae, usadas em minoria nos animais. As principais indicações de uso foram para o sistema digestório e tegumentar, seguido do musculoesquelético e respiratório, sob forma de infusão e decocto. Grande parte dos entrevistados desconhecia os efeitos adversos do uso inadequado de plantas medicinais, e espécies diferentes eram conhecidas com o mesmo nome popular, podendo resultar em toxicidade, dosagens errôneas e ações desconhecidas. Em relação ao cultivo e utilização das plantas medicinais, predominava em pessoas com idade média de 41,2 anos, que obtiveram o conhecimento principalmente de pais e avós. As plantas com potencial tóxico encontradas pertenciam às famílias Araceae, Euphorbiaceae e Araliaceae, totalizando 20 espécies; porém, apenas seis foram citadas como tóxicas pelos entrevistados. Assim, observou-se a existência e uso de muitas plantas medicinais, além da presença de espécies potencialmente tóxicas na região, sendo imprescindível ressaltar, com embasamento científico, possíveis benefícios e riscos da administração em animais.
The aim of this study was to identify plants used in traditional medicine, as well as those with toxic potential in the neighborhoods of Simões Lopes, Padre Réus and Porto in the city of Pelotas. For a year there were interviewed 111 owners of animals treated at the Veterinary Clinic-Universidade Federal de Pelotas, in order to assess the knowledge, cultivation, use and form of obtaining medicinal plants, age and sex of people, number of animals and results of the use of plants in the animals. After initial investigation, visits were held in the community aiming to shoot and collect samples of plants for botanical identification. They were listed 74 plants for medicinal purposes, mainly from Asteraceae and Lamiaceae families, minimally used in animals. The main indications of use were for digestive and integumentary system, followed by musculoskeletal and respiratory, by infusion and decoction. Most of the interviewers were unaware of the adverse effects of inappropriate use of medicinal plants, and different species were known with the same common name, so they could result in toxicity, wrong dosages and unknown actions. There was a predominance of cultivation and use of medicinal plants in people with an average age of 41.2 years who obtained knowledge mainly from parents and grandparents. The toxic potential plants found were from Araceae, Euphorbiaceae and Araliaceae families, in a total of 20 species, but only six were cited as toxic by the interviewees. There was noticed the existence and use of many medicinal plants and presence of potentially toxic species in the region; so it is essential to evidence scientifically possible benefits and risks of administration in animals.
Assuntos
Animais , Plantas Medicinais , Plantas Tóxicas , Asteraceae , LamiaceaeRESUMO
Antecedentes. El patógeno responsable de la pitiosis es Pythium insidiosum, un oomiceto acuático, perteneciente a la familia Pythiaceae, orden Peronosporales, clase Oomycetes, y perteneciente al reino Stramenopila. Objetivos. Describir la sintomatología, los cambios anatomopatológicos y métodos de diagnóstico de la pitiosis gástrica en perros. Métodos. Una perra hembra de raza Pastor Alemán, de 3 años de edad, y con acceso a tierras pantanosas, fue llevada a consulta por un cuadro de vómitos con diarrea recurrente de 30 días de evolución. Durante la exploración clínica, se identificó una masa palpable en el abdomen que ocupaba la región epigástrica izquierda, por lo que se procedió a la realización de un examen radiológico, simple y con contraste, junto con una ecografía abdominal. Más tarde, fue derivada para una laparotomía exploratoria con la finalidad de proceder a la exéresis de la masa. Debido a su tamaño, la exéresis no fue posible, por lo que se sacrificó al animal. Se obtuvieron muestras del tumor y se enviaron al laboratorio para su análisis morfológico e inmunohistoquímico. Resultados. Los cambios observados en los estudios de diagnóstico por imagen eran compatibles con un diagnóstico de pitiosis gástrica. En el examen citológico e histopatológico se identificaron hifas no tabicadas, mientras que el examen inmunohistoquímico fue intensamente positivo para la presencia de anticuerpos anti-Pythium, confirmando el diagnóstico de pitiosis. Conclusiones. El diagnóstico de pitiosis en perros se establece tardíamente, por lo que suele evolucionar hacia la muerte del animal. El diagnóstico definitivo debe establecerse a través de exámenes citológicos, histológicos e inmunohistoquímicos(AU)
Background. Pythiosis is caused by the agent Pythium insidiosum, an aquatic oomycete of the kingdom Stramenopila. Aims. To describe the symptoms, pathological changes and diagnosis methods of gastric pythiosis in dogs. Methods. A three-year-old female German shepherd, with access to wetlands, was attended due to vomiting and recurrent diarrhea of 30 days of duration. A palpable mass in the abdomen filling the left epigastric region was identified in the clinical examination. Simple and contrasted radiological examination and ultrasound of abdominal cavity were performed. The animal was referred for exploratory laparotomy for the removal of the mass. The extent of the mass prevented from the excision and the animal was euthanized. Samples of the tumor mass were collected and sent for morphological study and immunohistochemistry. Results. The changes observed in imaging studies were consistent with gastric pythiosis. In cytology and histopathology, non-septate hyphae were identified, and in immunohistochemistry a strong positivity of anti-Pythium antibodies was observed, confirming the diagnosis of pythiosis. Conclusions. Pythiosis in dogs is diagnosed late and tends to evolve in the animal's death. The definitive diagnosis is by cytology, histology and immunohistochemistry(AU)
Assuntos
Animais , Feminino , Cães , Pitiose/microbiologia , Pitiose/veterinária , Pythium/isolamento & purificação , Pythium/microbiologia , Diarreia/diagnóstico , Diarreia/veterinária , Laparotomia/métodos , Laparotomia/veterinária , Imuno-Histoquímica/métodos , Imuno-Histoquímica , Ultrassonografia , Vômito/complicações , Diarreia/complicações , Vômito/diagnóstico , Vômito/veterinária , Diagnóstico DiferencialRESUMO
BACKGROUND: Pythiosis is caused by the agent Pythium insidiosum, an aquatic oomycete of the kingdom Stramenopila. AIMS: To describe the symptoms, pathological changes and diagnosis methods of gastric pythiosis in dogs. METHODS: A three-year-old female German shepherd, with access to wetlands, was attended due to vomiting and recurrent diarrhea of 30 days of duration. A palpable mass in the abdomen filling the left epigastric region was identified in the clinical examination. Simple and contrasted radiological examination and ultrasound of abdominal cavity were performed. The animal was referred for exploratory laparotomy for the removal of the mass. The extent of the mass prevented from the excision and the animal was euthanized. Samples of the tumor mass were collected and sent for morphological study and immunohistochemistry. RESULTS: The changes observed in imaging studies were consistent with gastric pythiosis. In cytology and histopathology, non-septate hyphae were identified, and in immunohistochemistry a strong positivity of anti-Pythium antibodies was observed, confirming the diagnosis of pythiosis. CONCLUSIONS: Pythiosis in dogs is diagnosed late and tends to evolve in the animal's death. The definitive diagnosis is by cytology, histology and immunohistochemistry.