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69 RPCV (2013) 108 (585-586) 69-91 XVIII Meeting of the Portuguese Society of Animal Pathology - International Meeting of Leporidae Pathology Editorial The Portuguese Society of Animal Pathology (SPPA) is a small non-profit organization and a “daughter” So- ciety of the Portuguese Society of Veterinary Science (SPCV), which normally holds an annual meeting in the institutions from which its members belong (e.g. universities or government laboratories with activities in the field of Pathology). In an effort to decentralize their meetings, a tradition already started before, and both to aim at involving the community of teachers and students of the research in- stitutions that host the annual meeting and also to di- versify the main subject of its meetings, SPPA targeted new, younger and broader audiences, as well as new sectors and knowledge fields, trying to cover the pa- thology of a wide animal species range. Hence, to the present meeting we have chosen leporideae pathology as the main subject and a necropsy workshop, open to students and technicians - XVIII Meeting of the Por- tuguese Society of Animal Pathology. International Meeting of Leporidae Pathology, 9 and 10th of May 2013. Also, continuing the journey, already begun, of aiming evermore towards internationalization, this meeting will have reputable national and internation- al speakers, to whom above all I acknowledge their presence and immediate acceptance. Moreover, the scientific committee with national and international members, the three official languages, and the wid- est dissemination among technicians and students, will certainly make this meeting more open, more in- volved and more attractive. So we hope, so we wish, so we worked for! Our special guests will include an international ex- pert, who will share with us his scientific knowledge, Prof. Dr. Daniel Fernandez de Luco Martinez, profes- sor at the University of Zaragoza in the Department of Animal Pathology, Faculty of Veterinary Science. Graduated in Veterinary Medicine by the University of Zaragoza in 1985 and obtained a PhD by the same institution in 1991. He has developed his research in pathology of wildlife and wildlife hunting health con- trol in the region of Aragon in Spain. He has dozens of publications in international peer-review journals and was a guest speaker at dozens of conferences. He is responsible for several research projects, having juried several doctoral theses. Also, not forgetting nationals with high value and enriching practical experience in the field, an Europe- an expert, Dr. José Manuel Monteiro Azevedo. He has earned his degree in Veterinary Medicine in the Uni- versity of Tras-os-Montes and Alto Douro in 1999, and is currently a Veterinarian for Coren, Spain, being the Head of Industrial Rabbit Farming and the responsible for the Industrial Cuniculture in the Iberian Peninsula. His professional activity, since graduation, has been dedicated to rabbits having taken several positions at national and international companies. He collaborated in teaching several courses at the University of Trás- os-Montes and Alto Douro. We will have the opportunity to share and exchange experiences on animal pathology, results of research projects, discuss pathological diagnosis of difficult or rare cases, as well as the recent scientific advances in this field in Europe, which is undoubtedly the largest professional enrichment we could obtain. For the 18th time this small scale but dynamic meeting, constitutes an enrichment both scientifically, technically and pro- fessionally, and will allow us to grow a bit more pro- fessionally. Moreover, this will be also an excellent op- portunity to meet again our colleagues and friends and to discover Evora, one of the most interesting, beauti- ful and pleasant cities to visit in Portugal. The success of this meeting is already notorious by the number of members who decided to join us. Thank you for believing in this project, thank you for being here! In the name of SPPA and the Organizing Commit- tee, I want to acknowledge all those who contributed to make this event possible, namely the University of Évora, in the person of its Rector Ex º Sr. Prof. Dr. Carlos Alberto dos Santos Braumann and of its Vice- Rector, Prof. Dr. Marta Silvério, for immediately wel- coming this endeavor, namely for all the logistical sup- port provided and specially the generous availability in providing their facilities. I would like to acknowledge the Portuguese Society of Veterinary Sciences who provided an important assistance in the financial man- agement, and all those who helped us, teachers, stu- dents, staff and sponsors. The President of SPPA Helena Maria Vala Correia

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    RPCV (2013) 108 (585-586) 69-91

    XVIII Meeting of the Portuguese Society of Animal Pathology - International Meeting of Leporidae Pathology

    Editorial

    The Portuguese Society of Animal Pathology (SPPA) is a small non-profit organization and a daughter So-ciety of the Portuguese Society of Veterinary Science (SPCV), which normally holds an annual meeting in the institutions from which its members belong (e.g. universities or government laboratories with activities in the field of Pathology).

    In an effort to decentralize their meetings, a tradition already started before, and both to aim at involving the community of teachers and students of the research in-stitutions that host the annual meeting and also to di-versify the main subject of its meetings, SPPA targeted new, younger and broader audiences, as well as new sectors and knowledge fields, trying to cover the pa-thology of a wide animal species range. Hence, to the present meeting we have chosen leporideae pathology as the main subject and a necropsy workshop, open to students and technicians - XVIII Meeting of the Por-tuguese Society of Animal Pathology. International Meeting of Leporidae Pathology, 9 and 10th of May 2013.

    Also, continuing the journey, already begun, of aiming evermore towards internationalization, this meeting will have reputable national and internation-al speakers, to whom above all I acknowledge their presence and immediate acceptance. Moreover, the scientific committee with national and international members, the three official languages, and the wid-est dissemination among technicians and students, will certainly make this meeting more open, more in-volved and more attractive. So we hope, so we wish, so we worked for!

    Our special guests will include an international ex-pert, who will share with us his scientific knowledge, Prof. Dr. Daniel Fernandez de Luco Martinez, profes-sor at the University of Zaragoza in the Department of Animal Pathology, Faculty of Veterinary Science. Graduated in Veterinary Medicine by the University of Zaragoza in 1985 and obtained a PhD by the same institution in 1991. He has developed his research in pathology of wildlife and wildlife hunting health con-trol in the region of Aragon in Spain. He has dozens of publications in international peer-review journals and was a guest speaker at dozens of conferences. He is responsible for several research projects, having juried several doctoral theses.

    Also, not forgetting nationals with high value and enriching practical experience in the field, an Europe-an expert, Dr. Jos Manuel Monteiro Azevedo. He has earned his degree in Veterinary Medicine in the Uni-versity of Tras-os-Montes and Alto Douro in 1999, and is currently a Veterinarian for Coren, Spain, being the Head of Industrial Rabbit Farming and the responsible for the Industrial Cuniculture in the Iberian Peninsula. His professional activity, since graduation, has been dedicated to rabbits having taken several positions at national and international companies. He collaborated in teaching several courses at the University of Trs-os-Montes and Alto Douro.

    We will have the opportunity to share and exchange experiences on animal pathology, results of research projects, discuss pathological diagnosis of difficult or rare cases, as well as the recent scientific advances in this field in Europe, which is undoubtedly the largest professional enrichment we could obtain. For the 18th time this small scale but dynamic meeting, constitutes an enrichment both scientifically, technically and pro-fessionally, and will allow us to grow a bit more pro-fessionally. Moreover, this will be also an excellent op-portunity to meet again our colleagues and friends and to discover Evora, one of the most interesting, beauti-ful and pleasant cities to visit in Portugal.

    The success of this meeting is already notorious by the number of members who decided to join us. Thank you for believing in this project, thank you for being here!

    In the name of SPPA and the Organizing Commit-tee, I want to acknowledge all those who contributed to make this event possible, namely the University of vora, in the person of its Rector Ex Sr. Prof. Dr. Carlos Alberto dos Santos Braumann and of its Vice-Rector, Prof. Dr. Marta Silvrio, for immediately wel-coming this endeavor, namely for all the logistical sup-port provided and specially the generous availability in providing their facilities. I would like to acknowledge the Portuguese Society of Veterinary Sciences who provided an important assistance in the financial man-agement, and all those who helped us, teachers, stu-dents, staff and sponsors.

    The President of SPPA

    Helena Maria Vala Correia

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    Common gross lesions in farmed rabbits

    Fernndez de Luco D. *

    Departamento de Patologa Animal. Facultad de Vete-rinaria. Universidad de Zaragoza. C/ Miguel Servet 177. 50013 Zaragoza. Spain.

    * [email protected]

    Introduction: The identification of rabbit gross le-sions by systems and organs in association with a dis-ease, syndrome, aetiology or unknown cause is very important for veterinary practitioners.

    Material and methods: Necropsy has been made in several farmed rabbits with mainly respiratory or di-gestive problems, as well as delayed growth.

    Results: Dermatitis associated to myxomatosis, ringworm, mange or staphylococci are common.

    The respiratory system problems are localized in the nasal cavity and lungs mainly associated to Pasteurella multocida and Bordetella bronchiseptica. Young rab-bits are especially sensitive to acute and chronic pneu-monia. Haemorrhages can be seen in cases of rabbit calicivirus disease.

    Probably, digestive processes are the most important problems in growing rabbits. Epizootic rabbit enterop-athy, with still unknown aetiology, is a relatively recent process that has become the main cause of mortality in farmed rabbit. Other enteritis processes are princi-pally associated to coccidia, E. coli, Clostridium spp., C. spiroforme, C. piliforme (Tyzzers disease), Salmo-nella spp., as well as nematodes (Passalurus spp.). In-flammation and necrosis of the liver can be observed as small white spots associated to Eimeria stiedae (hepatic coccidiosis), Salmonella spp., C. piliforme, Cysticercus pisiformis, etc. Necrosis with karyorrhexis of the hepatocytes is a common finding in rabbit cali-civirus disease.

    Testis, uterus and mammary gland are the main or-gans affected in the reproductive system, as well as dystocia in pregnant rabbits. Processes and agents mainly involved are mixomatosis, syphilis (Treponema cuniculi), P. multocida, Staphylococcus spp., etc

    Finally, encephalitozoonosis by Encephalitozoon cuniculi is present in many rabbitries. Nervous signs, granulomatous encephalitis and interstitial nephritis are present in the affected animals.

    Situaes de urgncia em cunicultura industrial

    Monteiro J.M.1*, Alegria N.2, Coelho A.C.2

    1 Sumicor Portugal Lda; Coren S. C. G.2 Dept. de Cincias Veterinrias, CECAV, Universidade

    de Trs-os-Montes e Alto Douro (UTAD)

    * [email protected]

    Introduo: O mdico veterinrio durante a sua atividade contribui para a reduo das perdas associa-das morbilidade e mortalidade em cunicultura, para a melhoria dos resultados produtivos, atuando como consultor e orientando o produtor nos investimentos.

    O presente trabalho teve como objetivo reportar a atuao do mdico veterinrio na sua vertente clnica, dando nfase s principais situaes de emergncia em cunicultura.

    Material e mtodos: Procedeu-se realizao de um estudo observacional retrospetivo da actividade clnica na Pennsula Ibrica, durante os anos de 2009 a 2012.

    Foi efectuado o exame clnico dos animais prove-nientes de 76 exploraes, tendo-se procedido rea-lizao de necrpsias, com recurso a exames laborato-riais e elaborao dos respetivos registos que serviram de base ao presente estudo.

    Resultados: Determinou-se que a maioria das cha-madas de urgncia, se deveram s condies clnicas de enteropatia mucide, colibacilose, pasteurelose, po-dodermatite, mixomatose e enterotoxemia cujas leses anatomopatolgicas foram documentadas.

    As exploraes com maternidade e engorda no mes-mo pavilho, mau maneio, sem controlo ambiental e sujeitas a variaes climticas, situaes potencialmen-te geradoras de maior stresse e consequente reduo da imunidade, apresentaram aumento da morbilidade e menor produtividade, enquanto as exploraes com ambiente controlado e bom maneio, revelaram menos situaes de emergncia clnica e boa produtividade.

    Concluso: Foi possvel verificar que as exploraes onde as medidas de bem-estar animal e de controlo am-biental rigoroso j se encontravam implementadas, apre-sentaram melhores resultados produtivos e menor mor-bilidade em todas as fases de produo. Os resultados sugerem que a actuao do mdico veterinrio dever in-cidir tambm nas vertentes atrs referidas, pois s assim se conseguir aumentar a rentabilidade das exploraes.

    Emergency Situations in Industrial Rabbit Farming

    Introduction: The veterinary doctor in the course of his activities contributes to the reduction of losses as-

    Plenary sessions

  • SUPLEMENTO RPCV (2013) 108 (585-586) 69-91

    71

    sociated with morbidity and mortality, by improving the results of production, acting as a consultant, and orienting the producer in his investments.

    The present paper has as its objective to report on the performance of the veterinary doctor in the clini-cal aspect of his work, giving emphasis to the princi-pal emergency situations in rabbit farming.

    Material and methods: A retrospective observa-tional study was carried out of the clinical activity in the Iberian Peninsula during the years 2009 to 2012.

    The clinical examination of animals from 76 inves-tigations was carried out, subsequent to the realization of autopsies, with the use of laboratory tests whenever necessary and the elaboration of the respective regis-ters which served as a base for the present study.

    Results: It was found that the majority of emergency calls, resulted from the clinical conditions of mucoid enteropathy, colibacillosis, pasteurellosis, foot pad der-matitis, myxomatosis and enterotoxaemia, of which the

    anatomopathological injuries were documented.Farms with maternity and fattening stages in the same

    building, poor handling, without environmental control and subjected to climatic variations, all of which were potential generators of major stress and consequently reduction in immunity, showed an increase in morbidity and lower productivity, while the farms with environ-mental control and good handling revealed less clinical emergency situations and good productivity.

    Conclusion: It was possible to verify that the farms where the animal welfare measures being and rigor-ous environmental control had already been imple-mented showed better results in production and less morbidity in all the phases of production. The re-sults suggest that the activity of the veterinary doctor should also occur in the aspects previously referred; as it is only in this way that it will be possible to in-crease the profitability of the farms.

    Controlo de pragas em parques Zoolgicos: repercusses parasitrias, patolgicas e de Sade Pblica

    Crespo, A.P.1,2*, Lapo, N.1, Correia, J. 2, Gomes, L. 2, Vaz, Y.2 Madeira de Carvalho, L.M.2

    1 Jardim Zoolgico de Lisboa, Estrada de Benfica, 158-160, 1549-004 Lisboa;

    2 CIISA, Faculdade de Medicina Veterinria, Universi-dade Tcnica de Lisboa, Av. da Universidade Tcnica de Lisboa, 1300-477 Lisboa, Portugal ([email protected]);

    * [email protected]

    Introduo: A ligao prxima entre animais, Homem e pragas em zoos ou parques zoolgicos, constitui um risco na transmisso de muitos agentes de doena, alguns dos quais ocasionando doenas zoonticas, podendo afectar no s os animais da coleco zoolgica, como tambm trabalhadores do zoo e visitantes.

    Material e mtodos: Integrado num estudo so-bre controlo de pragas no Jardim Zoolgico de Lis-boa, efectuou-se um rastreio parasitolgico em 100 roedores (50 Mus musculus e 50 Rattus norvegicus). Os endoparasitas adultos foram coligidos aps de-cantaes dos contedos gastrintestinais e esfregaos por aposio do fgado (colorao pelo Giemsa) e de fezes (colorao pelo Ziehl-Neelsen), efectuando-se anlises coprolgicas por flutuao (mtodo de Wil-lis) e de sedimentao espontnea. Os fgados foram submetidos a anlise histolgica, fragmentos do f-

    gado foram fixados durante 24 horas, procedeu-se ao corte dos blocos de parafina e as amostras foram fixadas e coradas pela colorao da Hematoxilina e Eritrosina.

    Resultados: Dos roedores observados, 82 apresen-taram formas de endoparasitas, com prevalncias de 84,0% para R. norvegicus e de 80,0% para M. muscu-lus. Das espcies de roedores, Rattus norvegicus rep-resentou maior importncia na transmisso dos para-sitas assinalados. Identificaram-se um total de cinco espcies de endoparasitas com potencial zoontico: Hymenolepis diminuta (16,0%), Cysticercus fasciola-ris (12,0%), Syphacia obvelata (37,0%), Calodium hepaticum (32,0%) e Cryptosporidium sp. (5,0%).

    Concluso Deste conjunto de parasitas, C. hepati-cum, que j foi detectado aps a morte de primatas no humanos com consequncias patolgicas no f-gado daqueles hospedeiros. Alm da perda de ani-mais, o risco zoontico de transmisso de parasitas, bactrias e vrus aos trabalhadores e visitantes do Jardim Zoolgico de Lisboa, deve ser sempre con-siderado como um problema de grande importncia e impacto em Sade Pblica.

    Pest control in Zoos: parasitic, pathologi-cal and Public Health repercussions

    Introduction: The close connection between ani-mals, humans and pests, poses a risk for the transmis-sion of many agents, some of them causing zoonotic diseases, affecting not only the animals in the zoo-logical collection, as well as, zoo workers and visitors themselves.

    Oral communications

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    Material and methods: Following a pest control study at the Lisbon Zoo, a survey on endo and ectopar-asites was performed on 100 commensal rodents (50 Mus musculus and 50 Rattus norvegicus). Endopara-sites were collected from the gastrointestinal contents by a decanting process (adult specimens). Smears by liver apposition (Giemsa staining) and by extension feces (Ziehl-Neelsen staining), together with copro-logical exams by Willis flotation and spontaneous sedi-mentation techniques, were performed. The livers sub-jected to histological analysis of liver fragments were fixed for 24 hours, proceeded to cut the paraffin blocks and the samples were fixed and stained by Hematoxy-lin staining and Erythrosine.

    Results: From the total number of studied rodents, 82 were positive for endoparasites, with prevalence rates of 84.0% for R. norvegicus and 80.0% for M.

    musculus. Concerning the role of rodent species in parasite transmission, Rattus norvegicus represented the greater importance. Five species of endoparasites with zoonotic potential were identified: Hymenolepis diminuta (16.0%), Cysticercus fasciolaris (12.0%), Syphacia obvelata (37.0%), Calodium hepaticum (32.0%) and Cryptosporidium sp. (5.0%).

    Conclusion: Of this set of parasites, C. hepaticum constitutes an extreme example, since its presence was already detected after death in non-human primates, with severe consequences for these hosts liver. Beside the loss of animals, the risk of transmission of zoonotic parasites, bacteria and viruses to workers and visitors of the Lisbon Zoo should always be considered as a problem of great importance and impact on Public Health.

    Um caso de intoxico por cebola (Allium cepa) em bovinos

    Monteiro M. 1*, Mendona P. 1, Carvalho P.1, Maria-no I.2

    1 Laboratrio de Patologia/INIAV;2 Laboratrio Veterinrio de Montemor-o-Novo

    * [email protected]

    Introduo: A anemia hemoltica resultante do consumo prolongado de cebolas, Allium cepa, tem sido descrita em vrias espcies animais nomeada-mente em ces, bovinos e caprinos; o princpio txico presente na cebola o n-propil-dissulfito responsvel por alteraes da hemoglobina.

    Os Autores descrevem vrios casos de mortes em bovinos adultos que eram alimentados com pastagem natural e recebiam um suplemento de silagem de des-perdcio de tomate misturada com cebolas e alhos.

    Caso clnico: Os animais apresentavam anemia, hemoglobinria e em alguns casos, ictercia.

    Foram realizadas as necrpsias de dois animais; a abertura do cadver revelou um intenso cheiro a ce-bolas; observaram-se alteraes hepticas e renais.

    Discusso: Microscopicamente havia extensas zo-nas de necrose heptica centrolobular e necrose dos tubos renais simultaneamente com acumulao dum material compatvel com hemoglobina.

    Os exames efectuados em animais coabitantes, para diagnstico diferencial com doenas que podem apre-sentar sinais clnicos idnticos, resultaram negativos para leptospirose sendo detectada, em apenas um ani-mal, uma pequena infeco de Theileria annulata.

    Concluso: As leses microscpicas so sugestivas de uma anemia hemoltica tendo a sua associao ao suplemento alimentar em causa, permitido concluir

    que se trata dum caso de intoxicao por cebolas; acresce-se ainda que, aps ter sido retirado o referido suplemento e administrada uma teraputica de supor-te, os animais que estavam doentes recuperaram e no houve mais mortalidade.

    A case of onion (Allium cepa) intoxication in cattle

    Introduction: In domestic animals the prolonged ingestion of onion (Allium cepa) has long been asso-ciated with the emergence of hemolytic anemia. This toxicity is due to the presence of n-propil- disulphide.

    In this presentation the authors describe the occur-rence of several cases of toxicity in a grazing bovine herd due to ingestion of silage containing tomato, on-ion and garlic by-products.

    Clinical case: The affected animals showed ane-mia, haemoglobinuria and, occasionally, jaundice leading to death in some severe cases.

    Three animals were submitted to necropsy and the most prominent feature was a marked onion odor. The kidneys were dark brown and the liver slightly orange. Microscopically extensive areas of centrolobular liver necrosis as well as renal tubular necrosis with accu-mulation of hemoglobin-like material were seen.

    Differential diagnostic testing excluded leptopiro-sis and revealed one Theileria annulata infected ani-mal.

    Conclusion: The symptoms and the lesions ob-served were strongly suggestive of a haemolytic ane-mia. Taken together with the feed supplementation these facts enabled us to conclude for onion toxicity. Furthermore, after the discontinuation of the referred silage supplement and upon implementation of sup-portive care, the affected animals began to recover and no further deaths occurred.

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    Um caso raro de neoplasia retrobulbar

    Vala H1,2*, Mega A1,2, Santos C1,2, Cruz R1,2, Nbrega C1,2, Mesquita JR1,2, Esteves F1,2, Vieira F3, Ferraz A4

    1Escola Superior Agrria de Viseu, Instituto Politcni-co de Viseu, Portugal;2Centro de Estudos em Educao, Tecnologias e Sade, Instituto Politcnico de Viseu, Portugal;3Histocit, Portugal;4Clnica Veterinria Planeta Animal, Aveiro, Portugal.

    * [email protected]

    Introduo: Neoplasias envolvendo o nervo tico so raras. O meningioma intraorbitrio, a neoplasia mais comum sediada no nervo tico, constitui apenas 3% de todos os meningiomas em ces, razo pela qual existem poucos estudos para permitir prever os seus sintomas e comportamento biolgico.

    Objetivos: Os autores descrevem e submetem dis-cusso, um caso raro de meningioma retrobulbar, em particular para apreciao da graduao histolgica e o prognstico.

    Histria Clnica: Um Labrador Retriever de 10 anos de idade, foi apresentado com queixas de edema e descarga purulenta do olho direito. Apresentava exof-talmia, protuso da terceira plpebra, quemose, lcera profunda da crnea e ausncia de reflexo pupilar luz, no mesmo olho. O exame fsico, oftlmico e ecogr-fico permitiu diagnosticar exoftalmia devido a massa retrobulbar.

    Material e Mtodos: A amostra foi fixada em for-mol tamponado a 10 %, processada de acordo com o protocolo de rotina, tendo igualmente sido realizados estudos imunohistoqumicos, usando o mtodo do Complexo Estreptavidina-biotina-peroxidase.

    Resultados: O exame macroscopico revelou uma massa cnica, localizada na poro retrobulbar do ner-vo tico, adjacente poro posterior do globo ocular, apresentando colorao esbranquiada, aspeto homo-gneo, lobulado e consistncia firme.

    O exame microscopico revelou clulas fusiformes, com ncleo redondo a oval, cromatina em padro pon-tilhado, citoplasma eosinoflico, moderado, organizadas em ninhos, em torno de capilares, separados por feixes interlaados, com estroma muito vascularizado e escasso.

    Concluso: Os autores sugerem o diagnstico de meningiomado tipo transicional, tendo em conta as ca-ratersticas histopatolgicas e os resultados do estudo de imunohistoqumica.

    Agradecimentos: FCT e CI&DETS (PEst-OE/CED/UI4016/2011)

    A rare case of retrobulbar neoplasia

    Introduction: Neoplasms involving the optic nerve are rare. Intraorbital meningiomas, the most common tumour of the optic nerve, represent only 3% of all meningiomas in dogs, so few data exists to predict its symptoms and biologic behaviour.

    Objectives: The authors describe and submit for discussion, a rare case of retrobulbar meningioma, in particular for assessment of histological grade and prognosis.

    Medical History: A 10 year old Labrador Retriever male dog was presented for complaints of swelling and purulent discharge of the right eye. The animal had ex-ophthalmos, protrusion of the third eyelid, chemosis, deep corneal ulceration and no pupillary light reflex. Exophthalmos due to a retrobulbar mass was diagnosed after physical, ophthalmic and ultrasound examination.

    Material and Methods: The sample was fixed in 10 % buffered formalin solution, processed for routine histopathological diagnosis and a standard Streptavi-din-Biotin Complex technique for immunohistochemi-cal evaluation was also performed.

    Results: Macroscopic examination revealed a conic mass, located in the retrobulbar portion of the optic nerve, adjacent to the posterior aspect of the globe with homogeneous, lobulated and whitish appearance and firm consistency.

    Microscopic examination revealed spindled cells, with round-to-oval, finely stippled nuclei, small or indistinct nucleoli, moderate amount of eosinophilic cytoplasm, arranged in concentric whorls around cap-illaries, separated by interlacing fascicles, with small amount of well vascularized, collagenous stroma.

    Conclusion: The authors suggested the diagnosis of transitional meningioma, based on the histopathologi-cal diagnosis and immunohistochemistry results.

    Acknowledgments: FCT and CI&DETS (PEst-OE/CED/UI4016/2011)

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    Ocorrncia de parasitismo externo em coelhos de companhia

    Almeida, C.1*, Henriques C. 1, Bernardino, M. 1,2, Ma-deira de Carvalho, L.M. 2

    1 Exoclinic, R. Dom Antnio Ribeiro n1 loja B, 1495-145 Algs, Portugal

    2 CIISA, Fac Med Vet, Univ Tecn Lisboa, Av. da Univer-sidade Tcnica de Lisboa, 1300-477 Lisboa, Portugal

    * [email protected]

    Introduo: Em Portugal, tal como sucedeu ante-riormente noutros pases como Inglaterra, Estados Unidos e Alemanha, o coelho domstico tem revelado crescente popularidade como animal de companhia, devido sua grande inteligncia e interessante intera-o com a famlia.

    Durante o perodo de doze meses (Jan-Dez 2012) foram consultados 98 coelhos na Exoclinic sobre-tudo por problemas de sobrecrescimento dentrio, problemas gastrointestinais e manifestaes derma-tolgicas.

    Material e mtodos: Dos 25 casos de manifestaes cutneas, para alm do exame fsico foram submeti-dos a raspagem cutnea e citologia direta das leses. Foram identificados 16 coelhos positivos a parasitismo externo: Psoroptes cuniculi (69%) e Scarcoptes sca-biei (31%). Dos animais afetados, 81% apresentavam menos de seis meses de idade, verificando-se casos po-sitivos em machos e fmeas e nas raas Fuzzy-Lop, Lion Head e Mini Holands.

    O tratamento sugerido foi de injees subcutneas de ivermetina (200g/kg) repetindo ao fim de 14 dias, 28 dias, sugerindo uma quarta injeo e quinta injeo at desaparecimento total de sinais clnicos. Em alter-nativa, utilizou-se em metade dos casos, selametina so-luo tpica cutnea 15mg, repetindo-se ao fim de 28 dias, sugerindo uma terceira aplicao se necessrio.

    Resultados: Em termos de desaparecimento das le-ses cutneas e crescimento do plo nas zonas de alo-pcia os resultados obtidos foram bem sucedidos ao fim de 28 dias de tratamento e com eficcia equivalente com ambos os protocolos.

    Em simultneo foram aplicadas medidas de melhor maneio e higienizao das instalaes do animal, bem como o alerta do potencial zoontico de alguns tipos de sarna. Ainda assim, verificou-se a presena de dois episdios de manifestaes cutneas de sarna no pro-prietrio, j instaladas aquando do incio tratamento do animal.

    Concluso: De acordo com a nossa experincia, o clnico deve estar apto para a correta identificao dos agentes causais de problemas dermatolgicos em coelhos. O aumento da casustica destes pacientes re-presenta tambm novas perspetivas de diagnstico e

    novos desafios teraputicos. na nossa opinio uma oportunidade para instituir boas prticas veterinrias de preveno e profilaxia destas doenas, pois o bom estado geral de sade est condicionado pela correta informao e educao do proprietrio.

    Occurrence of external parasitism in pet rabbits

    Introduction: In Portugal, as happened previously in other countries like England, Germany and the United States, the domestic rabbit has shown increas-ing popularity as a pet, due to its great intelligence and interesting interaction with the family.

    During a twelve month period (Jan-Dec 2012) 98 rabbits were consulted in Exoclinic, mainly due to dental problems, gastrointestinal diseases and derma-tological manifestations.

    Material and methods: Of 25 cases of cutaneous manifestations, in addition to the physical examina-tion, they were subjected to scrape and direct cytol-ogy of cutaneous lesions. We identified 16 rabbits positive for external parasitism: Psoroptes cuniculi (69%) and Scarcoptes scabiei (31%). Of the affected animals, 81% were less than six months old, with pos-itive cases in both males and females and in Fuzzy-Lop, Lion Head and Mini Dutch breeds.

    The suggested treatment was subcutaneous injec-tions of ivermectin (200g/kg) repeating after 14 days and 28 days, suggesting a fourth and a fifth injection until total disappearance of clinical signs. Alterna-tively, in half the cases was used a topicalolution of selamectin 15mg, repeating the treatment 28 days af-ter the first one, suggesting a third application if nec-essary.

    Results: In terms of disappearance of skin lesions and hair growth in areas of alopecia, results were suc-cessful after 28 days of treatment, with equivalent ef-ficacy for both protocols.

    Simultaneously were applied measures for a better handling and hygiene of the animal facilities, as well as the alert for the zoonotic potential of some types of mange. Still, there was the presence of two episodes of cutaneous manifestations of scabies in the owners, already onset before initiating the animal treatment.

    Conclusion: According to our experience the clini-cian must be able to correctly identify the causative agents of skin problems in rabbits. The increase of these patients casuistic as pets also opens new pros-pects for diagnosis and new therapeutic challenges. It is in our opinion, an opportunity to establish good veterinary practice prevention and prophylaxis of these diseases, since the good general health is con-ditioned by proper education and information of the owner.

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    Identificao de Parasitas de Peixes em Cortes Histolgicos

    Ramos, P.1*, Carvalho, R.1, Rosa, F.2

    1 Laboratrio de Patologia dos Animais Aquticos. Ins-tituto Portugus do Mar e da Atmosfera, IPMA, I.P. Avenida de Braslia, 1449-006 Lisboa, Portugal

    2 Instituto de Investigao Cientfica Tropical, IICT. Rua da Junqueira n 14, 1300-343 Lisboa, Portugal

    1 J no se encontra a trabalhar na Instituio* Autor correspondente. Tel.: + 00351 213027000 Fax: + 00351 962467682

    * [email protected]

    Introduo: A identificao de parasitas de peixes em cortes histolgicos poder apresentar alguma difi-culdade, sobretudo quando as caractersticas morfol-

    gicas usadas na sua identificao no so observveis. Material e Mtodos: Apresentam-se trs casos de

    infeco parasitria: peritoneu de safio, Conger con-ger, com aspecto nodular; msculo de faneca, Trisop-terus luscus, com cor negra e corao de perca sol, Le-pomis gibbosus, envolvido por quistos de cor branca. Os tecidos foram fixados em formol salgado a 10% e processados histologicamente.

    Resultados: A estrutura nodular de peritoneu cor-respondia a um aglomerado de larvas enquistadas. A ausncia de progltides indicou tratar-se de espcimes imaturos. Nuns cortes, o parnquima larvar apresen-tava aspecto de tecido laxo, sem estruturas morfol-gicas e noutros, observou-se o esclex com botrdeas e quatro tentculos retrcteis armados com ganchos, bem como corpsculos calcreos correspondendo a larvas plerocercides de Trypanorhyncha. No mesmo tecido foram observadas larvas filiformes, com corpo

    Um caso de localizao exclusivamente gstrica de Spirocerca lupi numa raposa (Vulpes vulpes) em Portugal

    Silva J.F.*, Madeira de Carvalho L., Moreira A.S.

    CIISA - Faculdade de Medicina Veterinria (Universi-dade Tcnica de Lisboa), Av. da Universidade Tcnica (Polo Universitrio Alto da Ajuda), 1300-477 Lisboa

    * [email protected]

    Introduo: O Spirocerca lupi localiza-se caracte-risticamente na parede esofgica de Candeos. Ferran-telli et al.(2010) e Diakou et al.(2012), assinalaram a presena deste Nemtode na parede gstrica de raposas.

    Material e mtodos: O cadver de uma raposa adul-ta, sexo masculino, pesando 5,85 kg foi necropsiado e colhido material, que foi processado segundo a tcnica histolgica de rotina. A identificao dos Nemtodes em cortes histolgicos foi feita de acordo com o artigo de Chitwood e Lichtenfels (1972) e tendo em conta as-pectos bioecolgicos e epidemiolgicos.

    Resultados: Na parede do estmago observaram-se dois ndulos coalescentes, com dimetros respectiva-mente de 1,2 cm e 1,8 cm. Estes apresentavam super-fcie de corte branca, uma cavidade contendo material necrtico e um Nemtode enrolado sobre si prprio, de cor avermelhada, com cerca de 1 mm de espessura. O exame microscpico dos ndulos revelou uma cpsu-la de tecido conjuntivo fibroso envolvendo Nemtodes adultos. Estes apresentavam musculatura celomiria e polimiria, cordes laterais proeminentes, tero encer-rando ovos no operculados de casca fina, que conti-nham uma larva. O intestino apresentava um epitlio simples cbico com microvilosidades alongadas.

    Concluso: As caractersticas histolgicas, bioe-colgicas e epidemiolgicas do Nemtode permitem

    classific-lo como Spirocerca lupi, pelo que este um caso de localizao gstrica deste Nemtode em raposa em Portugal.

    A case of location exclusively in the gastric wall of Spirocerca lupi in a fox (Vulpes vulpes) in Portugal

    Introduction: One of the Spirocerca lupi current lo-cations is the oesophagic wall in Canidae. Ferrantelli et al. (201) and Diakou et al. (2012) reported the gastric wall location of this Nematode in foxes.

    Material and methods: An adult male fox weight-ing 5.85 kg was necropsied and samples were col-lected, which were processed according to the routine histological technique. The identification of Nematode specimens in histological sections was based in the pa-per by Chitwood and Lichtenfels (1972) and consider-ing bioecological and epidemiological aspects.

    Results: In the gastric wall were observed two coa-lescent nodules, which diameters were, respectively; 1.2 cm and 1.8 cm. Their cut surface was white, with a cavity containing necrotic material and a red Nema-tode coiled over itself, with 1 mm thick. The micro-scopic exam of the nodules revealed a capsule of dense connective tissue surrounding adult Nematoda. These worms possessed muscle cells of the coelomyarian and polymyarian types, prominent lateral chords, and an uterus containing thin-shelled, non-operculated ova, with a larva inside. The intestinal wall consisted of a simple cuboidal epithelium with long microvilli.

    Conclusion: The histological, bioecological, and epidemiological characteristics of this Nematode al-lowed us to classify it as Spirocerca lupi. The present case relates to an exclusively gastric location of this Nematode in a Portuguese fox.

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    circular em corte transversal, esfago de aspecto sim-trico radial e lmen triradial, correspondendo a larvas de nematodos.

    No msculo de faneca identificaram-se ovos bioper-culados, morulados ou castanhos embrionados, bem como pores de larvas ou adultos com corpo ciln-drico localizados entre as fibras musculares e dentro delas, resultando numa infeco parasitria por nema-todos Trichosomoididae, Huffmanella spp.

    No corao da perca sol, os quistos continham uma larva no segmentada, com dois cecos, uma ventosa oral e outra ventral, do trematode digentico, Postho-diplostomum minimum centrarchi.

    Concluso: Na identificao dos parasitas foi im-portante observar os caracteres morfolgicos, mas o conhecimento do peixe parasitado, o seu habitat e os tecidos afectados, auxiliaram o diagnstico.

    Identification of Fish Parasites in Stained Tissue Samples

    Introduction: In stained tissue sections prepared for light microscopy, taxonomic features may not be present, thus making parasite identification difficult. Three cases of parasite infection were described: Euro-pean conger, Conger conger peritoneum with nodular appearance; pouting, Trisopterus luscus with abnormal dark colour muscle and pumpkinseed sunfish, Lepomis gibbosus heart, surrounded by white colour cysts.

    Material and Methods: Tissues were fixed in 10% buffered neutral formalin and processed using standard methods.

    Results: The nodular peritoneum consists of multi-tude of encysted and clustered larvae. The absence of complete segmentation into proglottis identifies this specimen as immature. In sections, the body larvae vary from loosely parenchyma to prominent scolex features with bothridia and four tentacles armed with hooks. The larvae were identified as Trypanorhynchean plerocercoid. In this nodular peritoneum, filiform lar-vae were also present showing a round body surface in cross-section and the histological features of the oesophagus with its radial symmetry and triradiate lu-men which allowed classifying them as nematodes.

    The histopathological findings of the pouting mus-cle, revealed bioperculated eggs, immature and embry-onated brown eggs. The eggs and portions of larvae or adult nematodes Trichosomoididae of the genus Huff-manella were observed in either intracellular or extra-cellular locations.

    The heart of pumpkinseed sunfish was found infected by Posthodiplostomum minimum centrarchi larvae. The larvae were differentiated by the absence of the body segmentation and by an addition sucker, the ventral one.

    Conclusion: The morphological characters are im-portant in parasites identification but the knowledge on infected fish species, their habitat and infected tissues also provide additional information to the parasites identification.

    Caracterizao de um surto da nova variante da Doena Hemorrgica Viral (DHV) em coelhos bravos (Oryctolagus cuniculus algi-rus), Barrancos, Sudeste de Portugal

    Melo P.1*, Abrantes J.2,3, Ramada M. 1, Lopes A.M. 2,3,4, Alves P.C. 2,4, Esteves P.J. 2,5, Correia J.J. 6

    1 VETNATURA, Lisboa, Portugal *[email protected] CIBIO/UP, Centro de Investigao em Biodiversidade e Recursos Genticos/Universidade do Porto, InBio, Laboratrio Associado, Campus Agrrio de Vairo, Vairo, Portugal

    3 INSERM, U892, Universit de Nantes, Nantes, Fran-ce

    4 Departamento de Zoologia e Antropologia, Faculdade de Cincias da Universidade do Porto, Porto, Portugal

    5 CITS, Centro de Investigao em Tecnologias da Sa-de, IPSN, CESPU, Gandra, Portugal

    6 CIISA, Faculdade de Medicina Veterinria - Universi-dade Tcnica de Lisboa, Portugal

    Introduo: A doena hemorrgica viral (DHV) do coelho tem como agente etiolgico um calicivrus e causa elevada mortalidade em coelhos domsticos e

    bravos. Na Pennsula Ibrica, a DHV foi descrita em 1989 e todas as estirpes virais isoladas pertenciam ao genogrupo 1. Em 2011 detectou-se em Espanha uma nova variante deste vrus, descrita anteriormente em Frana, diferenciando-se por originar mortalidade em juvenis (

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    Principais dificuldades encontradas na aplicao das tcnicas de deteo de apoptose em tecidos animais

    Garcia C1*, Cruz R1,2, Mega A1,2, Oliveira AL3, Teixeira de Lemos E1,2,5, Ferreira D4, Reis F5, Vala H1,2

    1 Escola Superior Agrria de Viseu, Instituto Politcni-co de Viseu, Portugal;

    2 Centro de Estudos em Educao, Tecnologias e Sa-de, Instituto Politcnico de Viseu, Portugal

    3 Universidade de Lon, Lon, Espanha4 Laboratrio de Farmacologia e Teraputica Experi-mental, IBILI, Faculdade de Medicina da Universida-de de Coimbra, Coimbra, Portugal

    5 CICV- Centro de Investigao em Cincias Veterin-rias, Universidade Lusfona de Humanidades e Tec-nologia, Lisboa, Portugal

    * [email protected]

    Introduo: A aplicao de metodologias capazes de identificar clulas apoptticas constitui uma valiosa ferramenta em vrios estudos biomdicos. Muito do conhecimento adquirido acerca dos mecanismos regu-ladores da apoptose deriva de observaes em clulas isoladas. No entanto, as propriedades demonstradas por estes ensaios nem sempre se aplicam ao estudo de amostras tecidulares. Para deteo da apoptose em tecidos, muitos investigadores recomendam o mtodo

    TUNEL. Contudo, outros mtodos esto disponveis para avaliao da apoptose, devendo a sua escolha ser criteriosamente avaliada, tendo em conta as alteraes morfolgicas que se pretendem detetar. O presente tra-balho pretende, como principal objetivo, partilhar as dificuldades encontradas na implementao de dois diferentes mtodos para deteo de clulas apoptticas em tecidos animais.

    Material e mtodos: Foram utilizados tecidos de dife-rentes animais (rato, porco e co), fixados em formalina a 10% e includos em parafina, os quais foram submetidos a tcnicas imunohistoqumicas com recurso ao mtodo TUNEL (Kit In situ cell death, Roche) e aos anticorpos policlonais Bax e Bcl-2 (Santa Cruz Biotechnology).

    Resultados: Aps um longo perodo de testes e de custos, em que se obtiveram reaes inespecficas, bem como vrios artefactos - obstculos que, para serem superados, requerem persistncia, todos os mtodos provaram ser eficazes na deteo de apoptose, tendo sido as diferenas de marcao observadas relaciona-das com o tipo de tecidos testados.

    Concluso: Os mtodos imunohistoqumicos cons-tituem uma abordagem a considerar no que respeita deteo da apoptose em tecidos animais, sendo que as dificuldades encontradas representam um desafio su-pervel. Partilhar esta esperincia poder, certamente, evitar testes e custos desnecessrios sua implementa-o noutras unidades laboratoriais.

    Agradecimentos: FCT e CI&DETS (PEst-OE/CED/UI4016)

    A anlise filogentica demonstrou que as estirpes isoladas agrupam com a nova variante com elevado suporte estatstico.

    Concluses: Este estudo confirma a presena da nova variante em Portugal e levanta questes quanto conservao e sustentabilidade das populaes natu-rais de coelho bravo. Recomendam-se medidas adicio-nais de gesto nas unidades de criao.

    Characterization of an outbreak of the newly identified variant of rabbit haemor-rhagic disease virus (RHDV) in wild rabbits (Oryctolagus cuniculus algirus), Barrancos, Southeast of Portugal

    Introduction: Rabbit haemorrhagic disease virus (RHDV) is a calicivirus that causes a highly fatal dis-ease in wild and domestic adult European rabbits. In Iberian Peninsula, the first description of RHDV dates from 1989 and all isolated strains clustered within ge-nogroup 1. In 2011 a new variant of RHDV was detect-ed in Spain, already described in France, that differs by causing mortalities in young rabbits (

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    Otimizao da imunodeteo do recetor HER2 em tumores mamrios de cadela

    Beselga, A. 1, Soares, M. 1, Carvalho, S. 1, Matos, J. 2, Cabeadas, J. 2, Correia, J. 1, Peleteiro, M. 1, Ferreira, F. 1*

    1 CIISA, Faculdade de Medicina Veterinria, Univer-sidade Tcnica de Lisboa. Alto da Ajuda, 1300-477 Lisboa, Portugal

    2 IPOFG-EPE, Servio de Anatomia Patolgica, 1099-023 Lisboa, Portugal

    * [email protected]

    Introduo: O recetor para o fator de crescimento epidrmico de tipo 2 (HER2) apresenta-se sobrex-presso em 15-30% dos tumores mamrios na mulher, mostrando um elevado valor prognstico e teraputi-co. Na cadela, o papel oncognico do HER2 perma-nece duvidoso devido, em parte, falta de padroniza-o da tcnica de imunohistoqumica.

    Material e mtodos: Vinte e uma amostras de tumores mamrios caninos (TMC) foram colhidas e fixadas de acordo com as indicaes da American Society of Clinical Oncology (ASCO). Aps classi-ficao histopatolgica, as amostras foram sujeitas a diferentes protocolos imunohistoqumicos, tendo-se feito variar o mtodo de recuperao antignica e o anticorpo primrio usado: TAB250, diluio 1:250 (Zymed); SP3, 1:100 (Zytomed); CB11, 1:60 (Invi-trogen); 4B5, pronto a usar (Ventana). A interpreta-o dos resultados foi realizada de acordo com os critrios da ASCO.

    Resultados: Os melhores resultados foram obtidos

    com o anticorpo CB11 quando associado ao mto-do de recuperao antignica por panela de presso. Com este protocolo, 42,9% dos tumores analisados mostraram sobrexpresso do recetor HER2. Todos os anticorpos testados mostraram marcao membranar especfica no tecido mamrio de cadela, exceo do clone TAB250.

    Concluso: A variabilidade dos resultados obtidos demonstra a necessidade de padronizao da tcni-ca de imunohistoqumica para deteo do HER2 em Medicina Veterinria. Neste estudo, 42,9% dos TMC analisados mostraram sobreexpresso do HER2, um percentagem superior descrita na Mulher, reforan-do a possvel importncia deste biomarcador em On-cologia Veterinria.

    Optimization of HER2 receptor immuno-detection in canine mammary tumours

    Introduction: The epidermal growth factor type-2 receptor (HER2) is overexpressed in 15-30% of breast tumors in women, presenting a prognostic and high therapeutic value. In the bitch, the role of the HER2 oncogene remains doubtful due to the lack of immunohistochemistry technique standardization.

    Material and methods: Twenty-one canine mam-mary tumors (CMT) were collected and fixed in agreement to the American Society of Clinical On-cology (ASCO) guidelines. After histopathological classification, the samples were subjected to different immunohistochemical protocols, varying the recov-ery antigen method and the primary antibody used: TAB250, dilution 1:250 (Zymed); SP3, 1:100 (Zy-tomed); CB11, 1:60 (Invitrogen); 4B5, ready-to-use

    Main difficulties encountered in implemen-tation of techniques for apoptosis detection in animal tissues

    Introduction: The implementation of methodolo-gies capable of identifying apoptotic cells is a valu-able tool in many biomedical studies. Much of the knowledge acquired about the mechanisms regulat-ing apoptosis comes from observations in isolated cells; however, the properties shown by these assays do not always apply to the study of tissue samples. For detection of apoptosis in tissues, many investi-gators recommend the TUNEL method, nonetheless, other methodologies are available, and their choice must be carefully evaluated, taking into account the morphologic alterations which they intend to detect. The main goal of this work is to share difficulties encountered in the implementation of two different methods for the detection of apoptotic cells in animal tissue.

    Material and methods: Tissues from different ani-mals (rat, pig and dog) tissues, fixed in 10% neutral-buffered formalin and paraffin embedded were sub-mitted to immunohistochemical techniques using the TUNEL method (In situ Cell Death Kit, Roche) and polyclonal Bax and Bcl-2 antibodies (Santa Cruz Bio-technology).

    Results: After a long period of performed tests and expenses made, in which unspecific reactions, as well as several artifacts were obtained - obstacles that re-quired persistence to be overcome, all methods have proven to be effective in the detection of apoptosis, with differences probably related to the tissues tested.

    Conclusion: Immunohistochemical methods are a good approach to consider in the detection of apoptosis in animal tissues. Sharing the challenging difficulties found in our work, might obviate many tests and expens-es in its implementation in other laboratory facilities.

    Acknowledgments: FCT and CI&DETS (PEst-OE/CED/UI4016)

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    (Ventana). Results interpretation was performed ac-cording to the ASCO criteria.

    Results: The best results were obtained when the antibody CB11 was coupled with the pressure cooker as antigen retrieval method. With this protocol, 42.9% of the analyzed tumors showed HER2 overexpression. All antibodies tested show specific immunostaing of the cytoplasmic membrane, except the clone TAB250.

    Conclusion: The variability of the results obtained demonstrates the need of immunohistochemistry standardization for HER2 evaluation in Veterinary Medicine. In this study, 42.9% of TMC analyzed showed HER2 overexpression, a higher percentage than the one described in women, reinforcing the possible importance of this biomarker in Veterinary Oncology.

    Linfoma intestinal do tipo B com diferen-ciao em clulas de Mott num gato.

    Pedro Fasca1,2*; Maria Joo Costa3, Rita Pontes3, Dul-ce Carujo3, Matilde Gonalves4, Marli Angua1, Ins Carvalho2

    1 Faculdade de Medicina Veterinria da ULHT2 DNATech3 Clnica veterinria de Telheiras4 Servio de Anatomia Patolgica Centro Hospitalar de Setbal

    * [email protected]

    Introduo: Gato, macho, 11 anos de idade, que se apresentou consulta para vacinao anual. No exa-me fsico detetou-se massa abdominal, no dolorosa e firme.

    Materiais e mtodos: Realizou-se ecografia abdo-minal que confirmou massa intestinal.

    Posteriormente foi realizada a sua remoo por en-terectomia.

    Resultados: Macroscopicamente, ao nvel do jeju-no, identificou-se neoformao ulcerada, no esteno-sante, com 9x5x4,5 cm; em seco era branca amarela-da, brilhante, homognea, envolvendo toda a espessura da parede intestinal.

    Histologicamente observou-se proliferao difusa de clulas linfoides desde a mucosa at serosa. As-sociado aos linfcitos identificam-se inmeras clulas grandes e redondas, entre 20 a 30 m, com ncleo em-purrado para a periferia e citoplasma preenchido por glbulos eosinoflicos e hialinos, positivos com cido peridico de Schiff, compatveis com clulas de Mott. Os linfcitos apresentaram imunomarcao positiva para Pax5 (Leica NCL-L-PAX-5 1/80), CD79alphacy (Dako M7051, 1/50) e imunomarcao negativa para CD3 (Dako A0452, 1/50); as clulas de Mott demons-traram positividade para CD79alphacy, cadeias leves lambda (Ventana 760-2515) e negatividade para Pax5, CD3 e cadeias leves kappa (Ventana 760-2514). De-monstrou-se ndice mittico baixo (0-2 mitoses por campo com objetiva de 40x) e ndice de proliferao

    celular superior a 30% com o anticorpo Ki-67 (Dako M7240 1/50).

    Concluso: Com base nos achados morfolgicos, histoqumicos e imunohistoqumicos, o diagnstico histolgico compatvel com linfoma intestinal de tipo B, com diferenciao em clulas de Mott.

    Feline intestinal B-cell lymphoma with Mott cell differentiation.

    Introduction: An 11year-old, male cat, was present for the annual vaccination booster without any abnor-mal clinical signs. On clinical examination, a firm, non-painful abdominal mass was palpated.

    Materials and methods: An abdominal ecography confirmed the presence of an intestinal mass, which was posteriorly extirpated by enterectomy.

    Results: The jejunum presented a 9x5x4,5 cm, ho-mogenous, bright whitish-yellow mass surrounding the intestinal lumen and invading all intestinal layers.

    Histologically, there was a diffuse non-capsulated proliferation of round cells, disposed in dense sheets through all intestinal layers. The neoplastic cells were of lymphoid type associated with large (20-30um) round cells with cytoplasmatic eosinophilic, periodic acidSchiff positive, globules compatible with Mott cells.

    The lymphoid cells presented positive immunostain-ing for CD79alphacy (Dako M7051, 1/50), and Pax5 (Leica NCL-L-PAX-5 1/80), and negative immunos-taining for CD3 (Dako A0452, 1/50); the Mott cells presented positive immunostaining for CD79alphacy and lambda light-chain (Ventana 760-2515 ready to use), and were negative for Pax5, CD3 and kappa light-chain (Ventana 760-2514 ready to use). The mitotic figures were rare (02 per 40x field of view), and the Ki 67 proliferation index was higher than 30% (Dako M7240 1/50).

    Conclusion: On the basis of histopathology, histo-chemistry and immunohistochemistry, a B-cell lym-phoma with Mott cell differentiation was diagnosed.

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    Adenocarcinoma do endomtrio em gatas com menos de um ano de idade: descrio de quatro casos

    Pires M.A.1*, Saraiva A.L. 1,2, Santos T.3, Vilhena H.4, Sousa A.5, Santos C.1, Payan-Carreira R.1

    1 CECAV, University of Trs-os-Montes and Alto Dou-ro, P.O. Box 1013, 5001-801 Vila Real, Portugal.

    2 EUVG Escola Universitria Vasco da Gama, 3040-714 Castelo Viegas, Coimbra, Portugal.

    3 Veterinary Teaching Hospital (HVUTAD), University of Trs-os-Montes and Alto Douro, P.O. Box 1013, 5001-801 Vila Real, Portugal.

    4 Hospital Veterinrio do Baixo Vouga, 3750-742 gueda, Portugal

    5 Policlnica Veterinria de Aveiro - Vetsanus, 3800-118 Aveiro, Portugal.

    * [email protected]

    Introduo: As neoplasias uterinas de origem epi-telial so raras em todas as espcies animais, exceto em vacas e coelhas. Nas gatas, o adenocarcinoma do endomtrio (AEG) , geralmente descrito em fmeas com idade superior a 8 anos, ainda que, esporadica-mente, possa ser observado em idades compreendidas entre os 2 e os 4 anos.

    Material e mtodos: foram enviados ao Laboratrio de Histologia e Anatomia Patolgica da UTAD 4 peas de ovariohisterectomia (OVH) de gata, provenientes de diferentes Hospitais Veterinrios. Os rgos, fixa-dos em formol tamponado a 10% foram processados por rotina para parafina e corados com hematoxilina e eosina.

    Resultados: A sintomatologia observada foi vari-vel, desde a ausncia at presena de corrimento he-morrgico acastanhado,, facilmente confundidos com pimetra ou aborto.

    Macroscopicamente os teros apresentavam-se dis-tendidos, firmes, com parede espessada, dilataes e irregularidade dos cornos uterinos em diferentes graus. Microscopicamente observou-se proliferao papilar do endomtrio, com invaso do miomtrio em dois dos casos. O diagnstico foi de AEG de tipo papilar seroso.

    Concluso: Embora as neoplasias de origem epi-telial do tero sejam raras nos animais domsticos, o nosso grupo nos ltimos anos tem registado mais AEG

    do que o documentado. Embora esta neoplasia seja mais frequente em animais velhos, nos ltimos 2 anos tivemos 4 casos de AEG em gatas de idade inferior a um ano no momento da cirurgia.

    Esta situao clnica pode ser confundida com abor-to ou pimetra, pelo que se recomenda a anlise siste-mtica de todas as peas de OVH, seja qual for a idade ou diagnstico clnico.

    Feline Endometrial Adenocarcinoma in fe-males under one year-old: A description of four cases

    Introduction: Uterine neoplasms of epithelial origin are rare in all domestic animals, except for cows and rabbits. In female cats, endometrial adenocarcinoma (FEA) is usually described in females over 8 years-old. Yet, sporadically it may be found in younger females of age of 2-4 years.

    Material and methods: 4 ovariohysterectomy surgi-cal specimens from queens aged under one year old, at the time of surgery, were submitted from different practices to the Laboratory of Histology and Anatomi-cal Pathology, at UTAD. The organs were fixed in 10% buffered formalin, processed to paraffin embed tissue, and stained with haematoxylin and eosin.

    Results: Different clinical symptoms were observed in these animals, ranged from absent to a haemorrhagic or brownish discharge, easily confused with pyometra or abortion.

    Macroscopic outcomes revealed a distended and firm uterus, with wall thickness, dilatations and irregularity along both uterine horns, at different grades. Micro-scopic findings revealed a papillary proliferation of the endometrium with myometrium invasion in two cases. The final diagnosis was FEA of the papillary type.

    Conclusion: Although uterine neoplasms of epithe-lial origin are rare in domestic animals, in the last years our group has recorded more FEA than documented in the literature. Despite this neoplasia to be related with older animals, in less than two years, we untypically diagnosed 4 FEA in queens aged under one year-old at time of surgery that could be clinically mistaken for pyometra or abortion. For this reason, we strongly rec-ommend histopathological analysis of all ovariohys-terectomy surgical specimens, despite animals age or clinical diagnose.

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    Abate de Lagomorfos nos Matadouros Portugueses: Estudo Descritivo (2011-2012)

    Jesus J.1, Santos S.1*, Lamela M.2

    1 Centro de Investigao em Cincias Veterinrias

    (CICV) - FMV- ULHT, Lisboa, Portugal2 Chefe de Diviso na Direco de Servios de Segu-rana Alimentar da Direco Geral de Alimentao e Veterinria (DGAV)

    * [email protected]

    Introduo: Este trabalho d a conhecer os volumes de abate actuais de lagomorfos e as principais causas de reprovao sanitria nos matadouros portugueses.

    Material e mtodos: Analisou-se a totalidade de abates de lagomorfos entre 1 de Janeiro de 2011 e 31 de Dezembro de 2012, registados no Sistema de Infor-mao do Plano de Aprovao e Controlo dos Estabe-lecimentos (SIPACE), plataforma informtica da Dire-o Geral de Alimentao e Veterinria, utilizada por todos os inspetores sanitrios desde Outubro de 2010.

    Resultados: Foram apresentados para abate regular 10 901 526 lagomorfos. Desses, 321 (0,03) foram rejeitados ante-mortem e 93 665 (8,6) post-mortem. De entre os principais tipos de processos patolgicos observaram-se leses fibrino-purulentas, abcessos e caquexia. Registaram-se 20 497 mortes no transporte.

    Concluso: No perodo em anlise, o volume de abate manteve-se estvel e no apresentou sazonali-dade, sugerindo assim um consumo regular desta car-ne ao longo do ano. As leses fibrino-purulentas e os abcessos foram as principais causas de reprovao. A identificao das causas de reprovao fundamental

    para possibilitar a implementao de medidas preven-tivas.

    Slaughtering of Lagomorphs in the Portu-guese Slaughterhouses: Descriptive Study (2011-2012)

    Introduction: The present study was conducted to evaluate lagomorphss slaughtering data such as the current number of slaughtered animals and the major causes to condemnation in Portugal.

    Material and methods: This study includes the total number of lagomorphs slaughtered between 1st January 2011 and 31st December 2012. The data were obtained from Informatiom System of Establishments Approval and Control Plan (SIPACE) software, be-longing to Directorate General of Food and Veterinary (DGAV), where all veterinary inspectors insert the records since October 2010.

    Results: Out of 10 901 526 slaughtered lagomorphs, 321 (0,03) were rejected ante-mortem and 93 665 (8,6) were rejected post-mortem. The majority of injuries were fibrinopurulent lesions, abscesses and cachexia. Deaths in the transport accounted 20 497 animals.

    Conclusion: In the past two years the number of slaughtered lagomorphs remained stable and no sea-sonality in the number of slaughtered animals was noted, meaning that this type of meat is consumed regularly over the year. Fibrinopurulent lesions and abscesses were the most frequent causes of condemna-tion. Preventive actions can only be taken after identi-fication of the prevalent injuries, proving that studies like this are useful.

    Caracterizao da populao linfocitria em leses granulomatosas de javali (Sus scrofa) com diagnstico presuntivo de tu-berculose

    Matos A.C1., Andrade S.2, Pires M.A.2*, Coelho A.C.2, Pinto M.L.2

    1 Instituto Politcnico de Castelo Branco, Castelo Branco, Portugal

    2 CECAV - Centro de Cincia Animal e Veterinria; ECAV- Escola de Cincias Agrrias e Veterinrias, Vila Real, Portugal

    * [email protected]

    Introduo: A tuberculose doena infeciosa que ameaa a vida de milhes de pessoas, quer em pases

    em desenvolvimento, quer em pases desenvolvidos, sendo tambm responsvel por elevadas perdas econ-micas em animais de espcies pecurias. A patognese da tuberculose bovina, causada por Mycobacterium bo-vis e da tuberculose humana, cujo agente principal o microrganismo da espcie Mycobacterium tuberculo-sis, so as que se encontram melhor caracterizadas. No entanto, a tuberculose tambm afeta outras espcies de animais, sendo estes muitas vezes os responsveis pelo contgio s espcies pecurias e ao Homem. Em par-ticular, o estudo da tuberculose nas espcies selvagens pode elucidar a forma como esta doena se mantm e se transmite.

    Material e mtodos: Este trabalho foi realizado com o objetivo de avaliar a resposta inflamatria crnica em animais selvagens com diagnstico presuntivo de tuberculose, em que se procurou investigar a expres-so dos anticorpos anti-CD3 e anti-CD79 em leses

    Poster presentations

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    Estudo Preliminar sobre a Descalcificao de Tecido sseo em Micro-ondas

    Angua M.*, Fasca P.

    Centro de Investigao em Cincias Veterinrias- Fa-culdade de Medicina Veterinria - Universidade Lus-fona de Humanidades e Tecnologias

    * [email protected]

    Introduo: A anlise histopatolgica de tecido s-seo exige uma etapa de descalcificao, que possibilite a microtomia. O mtodo mais comum consiste na imerso das amostras em solues cidas, mas para alm de pro-vocarem danos tecidulares, o processo muito prolon-gado, o que atrasa significativamente o diagnstico. A utilizao de micro-ondas acelera a descalcificao, mas no deve comprometer a imagem microscpica.

    O objetivo deste trabalho consistiu na diminuio da durao da descalcificao, mantendo a qualidade da imagem microscpica.

    Material e mtodos: Foram testadas amostras de osso compacto e esponjoso, provenientes de caprinos, com dimenses que pretenderam simular casos de bi-psias e casos de peas cirrgicas ou necrpsias. Re-alizou-se a descalcificao pelo mtodo convencional

    e pelo mtodo em micro-ondas, atravs da adaptao de um protocolo conhecido. Utilizou-se cido ntrico a 5% e 10%.

    Resultados: Nos fragmentos de maiores dimenses, o protocolo foi interrompido aps 4 horas com cido ntrico a 10% em micro-ondas, por no se conseguir uma descalcificao completa e estar a ocorrer destrui-o do osso compacto. Nos fragmentos de osso espon-joso, verificou-se uma reduo de cerca de 25 horas relativamente ao mtodo convencional. Nas bipsias, houve uma reduo de aproximadamente 10 horas, uti-lizando cido ntrico a 5%. Com cido ntrico a 10% houve destruio tecidular. Nos casos em que se obteve uma descalcificao completa, a imagem microscpica apresenta fraca qualidade.

    Concluso: A utilizao de micro-ondas na descal-cificao com cido ntrico a 5% ou 10%, aplicando o protocolo deste estudo em amostras de caprinos, reduz a durao do processo, mas compromete significativa-mente a imagem microscpica, o que se torna invivel para diagnstico.

    Preliminary Study about Microwave De-calcification of Bone Tissue

    Introduction: Histological analysis of bone tissue

    granulomatosas dos gnglios mesentricos de javalis (n=30), pela tcnica de imuno-histoqumica.

    Resultados: Pela anlise imuno-histoqumica ob-servou-se marcao citoplasmtica das clulas lin-focitrias em 11 gnglios mesentricos (36,6%) para o anticorpo anti-CD79, e em 15 gnglios mesent-ricos (50%) para o anticorpo anti-CD3. A quantifica-o dessas mesmas clulas permitiu agrupar as leses consoante a percentagem de clulas linfocitrias que exibiam.

    Concluso: As leses em estudo apresentam percen-tagens relativamente semelhantes de linfcitos T e B, sugerindo assim uma resposta celular e humoral com igual participao na resposta imunitria, desencadea-da pelo organismo face entrada do agente patognico.

    Lymphocyte population in the granuloma-tous lesions of wild-boars (Sus scrofa) sus-pected of tuberculosis.

    Introduction: Tuberculosis is an infectious disease that threatens the lives of millions of people both in developing and developed countries and is responsible for important economical losses in farm animals. The pathogenesis of bovine tuberculosis due to Mycobac-terium bovis and human tuberculosis mainly caused by the microorganism of the species Mycobacterium

    tuberculosis are more extensively characterized. How-ever, tuberculosis also affects other animal species and these are often responsible for the contamination of cattle and humans. In particular, an overview of the wildlife hosts will provide insight into how these reser-voirs maintain and spread the disease.

    Material and methods: This study was conducted to evaluate the chronic inflammatory response in wild animals with a presumptive diagnosis of tuberculosis. In order to better understand the pathogenesis of the disease and further characterize its lesions, it was our aim to investigate the expression of antibodies anti-CD3 and anti-CD79 in the granulomatous lesions of wild boars mesenteric lymph nodes (n=30) through the immunohistochemistry technique.

    Results: Immunohistochemical analysis allowed us to observe the lymphocyte cells population in 11 me-senteric nodes (36.6%) for the anti-CD79 antibody, and in 15 mesenteric nodes (50%) for the anti-CD3 an-tibody. The quantification of these same cells allowed grouping of lesions depending on the percentage of lymphocyte cells that they exhibit.

    Conclusion: The lesions of this study showed rela-tively similar percentages of B and T lymphocytes, thus suggesting a cellular and humoral response and a similar immune response triggered by the organism against the entry of the pathogen.

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    Necrpsia em equinos. Descrio de um caso de doena granulomatosa disseminada

    Miranda I.*, Tilley P., Pomba C., Pissarra H., Correia J.J., Peleteiro M.C.

    Centro de Investigao Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinria, Univer-sidade Tcnica de Lisboa. Alto da Ajuda, 1300-477 Lisboa, Portugal

    * [email protected]

    Introduo: Tendo em conta a importncia de ava-liar a prevalncia das situaes ocorridas e a capacida-de de emisso de diagnsticos definitivos, procedeu-se identificao das causas de morte dos casos de ne-crpsia de cavalos registados no Laboratrio de Anato-mia Patolgica da Faculdade de Medicina Veterinria, da Universidade Tcnica de Lisboa. De entre os casos identificados foi dado particular relevo a um de doena granulomatosa disseminada (DGD), atendendo rari-dade do processo, complexidade da evoluo clnica e aos diagnsticos diferenciais envolvidos.

    Material e mtodos: Foram analisados os registos compreendidos entre Janeiro de 2009 e Janeiro de 2013, num total de 38 necrpsias. No caso da DGD, o diagnstico baseou-se na anlise histopatolgica de r-gos fixados em formol e submetidos a processamento histolgico de rotina. As coloraes efetuadas foram H&E e Ziehl Neelsen. Foi ainda realizada anlise bac-teriolgica.

    Resultados: Dos 39 casos de necrpsia analisados, apenas um foi inconclusivo. Os restantes foram divi-didos em seis grupos: Doena Infeciosa Sistmica (5), Doena Cardiorrespiratria (5), Doena Renal (1), Do-ena Gastrointestinal (21), Doena Neurolgica (3), Doena Ortopdica/Trauma (3). Os exames comple-mentares realizados foram anlise histopatolgica (24) e anlise bacteriolgica (10).

    O diagnstico de doena granulomatosa dissemina-da compatvel com sarcoidose equina baseou-se nas le-ses identificadas na necrpsia e na anlise histopato-lgica de leses granulomatosas presentes em diversos rgos, ricas em clulas gigantes de Langhans e negati-vas para a presena de bacilos lcool-cido resistentes.

    Concluso: Como concluso, pode referir-se que re-alizao de necrpsia permite, na grande maioria dos casos, identificar a causa da morte em equinos, pre-ferencialmente com recurso a outros exames comple-mentares de diagnstico.

    Necropsy in horses. A case of disseminated granulomatous disease

    Introduction: With the objective of estimating the prevalence of the occurrences and the capability to is-sue definitive diagnosis, an evaluation of the various causes of death in horses was performed based on the necropsy cases registered in the Laboratory of Pathol-ogy, Faculty of Veterinary Medicine, Technical Univer-sity of Lisbon. Among the cases identified, particular emphasis was given to a case of disseminated granu-lomatous disease (DGD), due to the rarity of the pa-thology, complexity of the clinical evolution and the differential diagnosis involved.

    Material and methods: The reports from January 2009 to January 2013 were analyzed, and a total of 39 necropsies recovered. In the case of DGD the conclu-sion was based on the histopathology of the organs fixed in 10% formalin and subject to regular histologi-cal processing. H&E and Ziehl Neelsen stains were performed. Bacteriology was also performed.

    Results: Out of 39 cases of necropsy analyzed, only one was inconclusive. The remainders were divided into six groups: Systemic Infectious Disease (5), Car-diopulmonary Disease (5), Kidney Disease (1), Gas-trointestinal Disease (21), Neurological Disease (3) and Trauma/Orthopedic Disease (3). Complementary

    requires a decalcification process. The most common method consists on samples immersion in acidic so-lutions. However, it causes tissue damage and is time consuming, delaying the diagnosis. The use of mi-crowaves accelerates the decalcification process, but shouldnt compromise the histological image.

    The aim of this work was to reduce the decalcification time, maintaining the quality of the histological image.

    Material and methods: Goat compact and spongy bone, with dimensions that intended to simulate bi-opsies and surgical or necropsies specimens, were used. Decalcification was performed by conventional method and microwave method with an adaptation on a known protocol, using 5% and 10% nitric acid.

    Results: For larger fragments, the protocol was dis-

    continued after 4 hours of microwave cycles** with 10% nitric acid, because the decalcification was in-complete, in spite of evident compact bone destruc-tion. In the fragments of spongy bone, there was a reduction of approximately 25 hours, comparing with conventional method. In biopsies, there was a reduc-tion of approximately 10 hours using 5% nitric acid. With 10% nitric acid, there was tissue destruction. The cases with complete decalcification had a poor quality of the histological image.

    Conclusion: The use of microwave decalcification with 5% or 10% nitric acid, applying the protocol of this study on goats samples, reduces the process time but compromises the histological image, which is im-practical for diagnosis purpose.

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    Processos patolgicos prevalentes em carnvoros silvticos mortos em acidentes rodovirios

    Matos A.C1., Figueira L1., Martins M.1, Matos M.2, l-vares S.1, Mendes A.1, Pires M.A.3, Coelho A.C.3, Pinto M.L.3*

    1 Instituto Politcnico de Castelo Branco, Castelo Branco, Portugal

    2 Centro de Gentica e Biotecnologia, Instituto de Bio-tecnologia e Bioengenharia, Universidade de Trs-os-Montes e Alto Douro, Vila Real, Portugal

    3 CECAV - Centro de Cincia Animal e Veterinria; ECAV- Escola de Cincias Agrrias e Veterinrias, Universidade de Trs-os-Montes e Alto Douro, Vila Real, Portugal

    * [email protected]

    Introduo: As doenas que afetam os animais selvagens tm sido alvo de um interesse crescente,

    uma vez que se demonstrou que, para alm das impli-caes relacionadas com a conservao das espcies, podem ter um impacto significativo na sade animal e humana. Devido ao seu comportamento furtivo, os carnvoros selvagens constituem uma populao difcil de monitorizar e a investigao de doena nestas es-pcies resulta geralmente do exame de animais que so encontrados mortos.

    Material e mtodos: Este estudo foi realizado em carnvoros selvagens vtimas de acidentes de viao, em Portugal, entre 2009 e 2012, e inclui 3 famlias e 6 espcies diferentes (n=72). Os animais foram submeti-dos a exame post-mortem e foram efetuadas colheitas, sendo os tecidos fixados em formol tamponado a 10%, processados para incluso em parafina e corados com hematoxilina-eosina.

    Resultados: As leses pulmonares foram as mais prevalentes, (15/72; 20,8%), especialmente em ra-posas (Vulpes vulpes, 8/49; 16,3%) e saca-rabos (Herpestes ichneumon, 7/15; 46,6%). As pneumonias parasitrias, sugestivas de infestao por Crenosoma vulpis ou Angiostrongylus vasorum foram observadas

    Fibromatosis in Iberian hare Lepus granat-ensis. Preliminary data.

    Arnal MC, Martnez-Durn D, Revilla M, Fernndez de Luco D *

    Departamento de Patologa Animal. Facultad de Vet-erinaria. Universidad de Zaragoza. C/ Miguel Servet 177. 50013 Zaragoza. Spain.

    * [email protected]

    Introduction: Hare fibromatosis is a rare and trans-missible viral disease, caused by a Leporipoxvirus, antigenically more related to the Shope fibroma virus than myxoma virus. The disease is characterized by single or multiple skin tumours on ears, head and legs. The affected animals appear in late summer and au-tumn with high morbidity and low mortality. Fibroma-tosis has been described in wild and farmed brown hare Lepus europaeus in Europe (France, Germany, Italy, U.K.). African hare fibromatosis is a very similar disease caused by an unclassified Poxvirus affecting Cape hare Lepus capensis in Kenya. Recently, another

    similar disease has been reported in mountain hare Le-pus timidus in Finland, where gross and microscopic lesions resembled to be associated to a Poxvirus infec-tion.

    Material and methods: We describe here two natu-ral cases in wild Iberian hares Lepus granatensis in Aragon region (Spain). Both hares were adults female and male hunted in the autumn period in 1998 and 2006, respectively.

    Results: Noticeable swelling eyelids were observed in both animals and the male also showed small nodu-lar lesions in nose and lips, swelling prepuce and an irregular swelling of scrotum skin. The dermis histopa-thology of skin gross findings consisted in fibroblast cells proliferation and oedema, whereas the epidermis showed swelling and vacuolated cells with intracyto-plasmic eosinophilic inclusion bodies. Infiltration of plasma cells and lymphocytes were observed in the dermis. Neutrophils were present in the dermis and epidermis, probably by secondary bacterial infections.

    Conclusion: The observed lesions could be associat-ed to hare fibromatosis in Iberian hare Lepus granat-ensis, in Spain.

    exams were histopathological examination (24) and bacteriological analysis (10).

    The diagnosis of disseminated granulomatous dis-ease consistent with equine sarcoidosis was based on the gross morphology at necropsy and histopathology of granulomatous lesions with Langhans giant cells

    present in different organs that were negative for the presence of acid-fast bacilli.

    Conclusion: It can be concluded that, in most cases, necropsy allows for the identification of the cause of death in horses, preferentially using other complemen-tary exams to achieve a correct and complete diagnosis.

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    Doena vrica hemorrgica (DVH) uma nova variante?

    Vala H1,2*, Coelho C1,2, Nbrega C1,2, Mesquita JR1,2, Oliveira J1,2

    1 Escola Superior Agrria de Viseu, Instituto Politcni-co de Viseu, Portugal;2 Centro de Estudos em Educao, Tecnologias e Sa-de, Instituto Politcnico de Viseu, Portugal

    * [email protected]

    Introduo: A doena vrica hemorrgica (DVH), causada por um RNA vrus (gnero Lagovrus, famlia Caliciviridae), uma doena fatal que afeta coelhos do-msticos e selvagens. Estudos anteriores a 1984, demons-traram a existncia de anticorpos especficos para DVH em coelhos saudveis de vrias regies europeias e ainda a possibilidade de estes animais apresentarem alguma re-sistncia contra a DVH conferida por uma imunidade cru-zada com calicivrus benignos, fazendo com que a doena exibisse um quadro clnico menos severo. A doena foi controlada em cuniculturas graas vacinao, mas ain-da uma ameaa nas populaes selvagens no vacinadas.

    Uma nova variante desta doena, descrita em 2010 na Pennsula Ibrica, afeta principalmente animais jo-vens, que exibem hemorragias no corao, traqueia, timo, pulmes, fgado, rins, intestino, assim como

    CID, ictercia, esplenomeglia e enterite catarral.Objetivos: Descrever um surto de elevada morta-

    lidade numa cunicultura, principalmente em animais jovens, com leses tpicas de DVH.

    Material e mtodos: Foram colhidas amostras de co-elhos vacinados e no vacinados, alvo de morte sbita. As amostras foram fixadas em formalina a 10% (48 ho-ras), embebidas em parafina e processadas seguindo os procedimentos de rotina para avaliao histopatolgica

    Resultados: Principais leses observadas: opist-tonus; congesto e edema pulmonar; ictercia severa; esplenomeglia; lceras gstricas punctiformes; con-gesto renal bilateral, petquias (timo).

    Concluso: Embora esta infeo seja mais comum em animais adultos, as leses observadas so consis-tentes com DVH. A presena concomitante de leses associadas nova variante de DVH (CID, ictercia, esplenomeglia, enterite catarral) leva-nos a suspeitar da sua presena em Portugal, ressalvando o facto da necessidade de estudos mais aprofundados.

    Agradecimentos: FCT e CI&DETS (PEst-OE/CED/UI4016/2011)

    Rabbit haemorrhagic disease (RHD) a new variant?

    Introduction: Rabbit haemorrhagic disease (RHD) is an acute fatal disease of domestic and wild Euro-

    em 6 raposas. A adiaspiromicose, provocada por Em-monsia crescens, foi a leso pulmonar mais comum em saca-rabos (4/15, 26,6%). A nefrite intersticial crnica e a linfadenite macrofgica afetaram, respetivamente, 10/72 (13,8%) e 6/72 (8,3%) animais das famlias Ca-nidae e Mustelidae.

    Concluso: De acordo com este estudo, as pneu-monias parasitrias e micticas encontram-se entre as leses mais prevalentes em carnvoros selvagens e podem ter um impacto significativo na sade destas es-pcies, quer devido magnitude das leses observadas, quer devido ao aumento da susceptibilidade a outros agentes infeciosos.

    Disease processes identified in road killed wild carnivores

    Introduction: Wildlife diseases have been considered increasingly important due to their potential effect upon human health, veterinary medicine, wildlife, and con-servation biology. Since wild carnivores are elusive ani-mals, the occurrence of disease in these species has been typically investigated by identification of pathological processes in carcasses that are found incidentally.

    Material and methods: This study was conducted in wild carnivores found death on the roads in Portu-

    gal, from 2009 to 2012 (n=72), representing 3 families and 6 different species. Post mortem examination was performed and multiple tissues were fixed in 10% neu-tral buffered formalin, processed for routine paraffin embedding and stained with hematoxylin and eosin for histopathological examination.

    Results: Lung lesions were the most prevalent find-ing (15/72, 20.8%), particularly in fox (Vulpes vulpes, 8/49, 16.3%) and Egyptian mongooses (Herpestes ichneumon, 7/15, 46.6%). Six foxes showed lesions of parasitic pneumonia suggestive of Crenosoma vul-pis or Angiostrongylus vasorum infestation. In Egyp-tian mongooses adiaspiromycosis due to Emmonsia crescens were the most common lung lesions (4/15, 26.6%). Chronic interstitial nephritis was registered in 10/72 (13.8%) animals, while in 6/72 (8.3%) macro-phage lymphadenitis was observed, both conditions af-fecting members of the Canidae and Mustelidae.

    Conclusion: According to our study, parasitic and mycotic lung pneumonias are amongst the most preva-lent lesions in wild carnivores. Although parasites and fungus may often be regarded as a burden that has little or no impact on the animals, our findings suggest that they can severely affect animal health by the magni-tude of their lesions and by making turning the host more susceptible to other infectious agents.

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    Timo de Coelho um potencial marcador para tcnicas de deteo de apoptose

    Vala H1,2*, Santos C1,2, Cruz R1,2, Nbrega C1,2, Ortiz AL3, Mesquita JR1,2, Esteves F1,2, Mega A1,2,

    1 Escola Superior Agrria de Viseu, Instituto Politc-nico de Viseu, Portugal;

    2 Centro de Estudos em Educao, Tecnologias e Sade, Instituto Politcnico de Viseu, Portugal

    3 Len University, Len, Spain

    * [email protected]

    Introduo: As tcnicas de determinao de apop-tose tm sido preconizadas para fins de diagnstico, designadamente para determinao das caratersticas biolgicas das neoplasias e, indiretamente, determi-nao do prognstico.

    A implementao de uma nova tcnica num labo-ratrio pressupe o uso de um controlo positivo efi-caz para aferio da mesma. Muitas das publicaes da especialidade sobre o tema no indicam, de modo claro, os controlos positivos usados e no divulgam os resultados dos mesmos.

    O objetivo deste trabalho consistiu na implemen-tao de trs mtodos de deteo de apoptose, utilizan-do trs amostras de tecidos de co: neoplasia mamria - carcinoma in situ, timo e plasmocitoma; e uma amos-tra de tecidos de coelho timo.

    Material e mtodos: Os mtodos de imunohis-toqumica incluram a deteo de protenas pro- e anti-apoptticas, respetivamente, Bax e Bcl-2 (1:100; Santa Cruz, Biotechnology) e o mtodo de TUNEL, com o kit de deteo de apoptose in situ (1:2,5; Roche).

    Resultados: O plasmacitoma revelou reao posi-tiva moderada com todos os mtodos testados. O car-

    cinoma in situ revelou reao positiva moderada com os anticorpos pro-apoptticos e mais intensa com o anticorpo anti-apopttico. O timo de co revelou reao positiva moderada nos timcitos do crtex com os anticorpos pr-apoptticos e dbil nos da medular. Com o anticorpo anti-apopttico revelou reao dbil na medular e negativa no crtex. Idnticos resultados foram obtidos no timo de coelho, tendo sido, contudo, mais forte a intensidade das respetivas reaes.

    Concluso: O timo de coelho revelou a reao mais especfica dos tecidos estudados, sendo assim reco-mendado como um bom controlo positivo para estudos de apoptose.

    Agradecimentos: FCT e CI&DETS (PEst-OE/CED/UI4016/2011)

    Rabbit thymic tissue a potential marker for apoptosis techniques

    Introduction: Apoptosis methods have been pro-posed for diagnostic purposes, in particular for deter-mining the biological characteristics of tumours and, indirectly, their prognosis.

    Implementation of a new technique requires the use of an effective positive for correct assessment.

    Most specialty publications on the subject, do not indicate the positive controls used and do not disclose the obtained results therein.

    Objective: The aim of this work was to implement three different methods for the detection of apoptosis, using three canine tissue samples, consisting of breast cancer - carcinoma in situ, thymus and plasmacytoma; and a sample of rabbit tissues - thymus.

    Material and methods: Immunohistochemistry was performed, using pro- and antiapoptotic proteins, respectively, Bax and Bcl2 (1: 100; Santa Cruz, Bio-

    pean rabbits caused by a RNA virus (genus Lagovirus, family Caliciviridae). Prior to 1984, healthy rabbits in European regions had RHDV-specific antibodies and their infection with benign rabbit caliciviruses con-ferred them a low degree of cross-protective immunity against RHD, determining less severe clinical courses. Vaccination allowed to control the disease in the rab-bit industry, but RHD still threatens non-vaccinated wild populations. A new variant of this disease is be-ing described since 2010 in Iberian Peninsula. It af-fects mainly young rabbits that exhibit haemorrhages in heart, trachea, thymus, lungs, liver, kidneys and gut, as well as DIC, jaundice, splenomegaly and catarrhal enteritis.

    Objectives: To describe a case of high mortality rate on a rabbitry, mostly in young rabbits, with typical le-sions of RHD.

    Material and methods: Samples from domestic

    vaccinated and non vaccinated rabbits which died sud-denly were taken and fixed in 10% neutral-buffered formalin for 48 hours and embedded in paraffin. 3m sections were stained for routine histopathological di-agnosis (H&E) and examined by light microscopy.

    Results: Main macroscopic lesions observed: opisthotonos, oedema and pulmonary congestion, se-vere jaundice, splenomegaly, gastric punctiform ul-cers, bilateral renal congestion and thymic petechiae,

    Conclusion: Macroscopic lesions in infected youngs, consistent with RHD infection usually ob-served only in adult rabbits, and the presence of le-sions mostly associated to the new variant of RHD (DIC, jaundice, splenomegaly, catarrhal enteritis), led us to consider the presence of the new variant of RHD in Portugal, although further studies are needed.

    Acknowledgments: FCT and CI&DETS (PEst-OE/CED/UI4016/2011)

  • SUPLEMENTO RPCV (2013) 108 (585-586) 69-91

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    Um caso de hemangiosarcoma cardaco em co

    Pires M.A.1,2*, Pinheiro D. 1, Silva N.3, Montenegro L.3

    1 CECAV, University of Trs-os-Montes and Alto Dou-ro, P.O. Box 1013, 5001-801 Vila Real, Portugal.

    2 ECAV, University of Trs-os-Montes and Alto Douro, P.O. Box 1013, 5001-801 Vila Real, Portugal.

    3 Veterinary Montenegro Hospital (HVM), 4200 - 447 Porto, Portugal.

    * [email protected]

    Introduo: Segundo a literatura, o hemangiossar-coma cardaco uma situao comum nos ces Pastor Alemo, comumente associado a leso esplnica.

    Os autores descrevem um caso de hemangiossarco-ma cardaco sem envolvimento esplnico, com par-metros clnicos e imagens macroscpicas, associado a metstases pulmonares.

    Material e mtodos: Foram remetidos ao Laborat-rio de Histologia e Anatomia Patolgica da UTAD, o corao e pulmo de um Pastor Alemo, macho, de 11 anos de idade. Os rgos fixados em formol tampona-do a 10%, foram processados por rotina e corados com hematoxilina e eosina.

    Resultados: O historial do animal indicava ausncia de vacinao e desparasitao, fraqueza extrema, disp-neia e choque. Ao RX a silhueta cardaca apresentava-se arredondada e com imagens de efuso torcica e pericrdica, confirmada por ecografia.

    Foi solicitada a eutansia, tendo a necrpsia revelado hemotrax e hemopericrdio, leso papilar exofitica na aurcula direita, vilosa, de 3 cm de dimetro que inva-dia o miocrdio, at ao pericrdio com 6x3x4,5cm. O pulmo apresentava mltiplas pequenas leses redondas e negras, a maior com 0,9 cm de dimetro. No se ob-servaram outras leses macroscpicas dignas de registo.

    O corao apresentava proliferao de clulas fuso-celulares, com elevado pleomorfismo e clulas atpicas, com 4 a 5 mitoses/40x, com disposio estoriforme e fendas com eritrcitos. O pulmo apresentava mlti-plas metstases com clulas e disposio semelhantes ao observado no corao, algumas intimamente ade-

    rentes aos vasos sanguneos. Todo o tecido adiposo ad-jacente ao corao apresentava proliferao de clulas neoplsicas.

    Concluso: O diagnstico foi de hamangiossarcoma da aurcula direita com metstases pulmonares, sem envolvimento esplnico.

    A case of heart hemangiosarcoma in a dog

    Introduction: The consulted literature, referred that hear hemangiosarcoma, a common condition in Ger-man Shepherd dog, generally associated with spleen hemangiosarcoma.

    The authors report a case of heart hemangiosarcoma without spleen implication, with clinical parameters and macroscopic images, associated with lung metastasis.

    Material and methods: an heart and lung from an eleven years old male German Shepherd dog was to the Laboratory of Histology and Anatomical Pathology, at UTAD. The organs were fixed in 10% buffered formalin, and processed and stained with haematoxylin and eosin.

    Results: The clinical history indicated absence of vaccination or dewormed medication, generalized weakness, dyspnoea and shock. At x-ray the cardiac silhouette is rounded with pleural and pericardial effu-sion confirmed by ultrasonography.

    Euthanasia was requested and at necropsy an abun-dant hemothorax, and hemopericardium was recorded. The right auricle presents a papillary villous exofitic proliferation with 3cm of diameter, that invaded the myocardium until the pericardial wall with overall dimensions 6x3x4.5cm. The lung presented multiple black round nodules, the larger with 0.9 cm diameter. There were no other gross lesions noteworthy.

    The histology of the heart lesion revealed fusocel-lular proliferation with high pleomorphic and atipic cells, with 4-5 mitosis for HPF. The neoplasia had a storiform pattern with clefts with erythrocytes. The lung had multiple metastases with similar cells, some of them within vascular structures. All adipose tissue around the heart had neoplasic cells.

    Conclusion: The final diagnosis was hemangiosar-coma of the right auricle with lung metastasis, without spleen implication.

    technology) and the TUNEL technique (1: 2,5; Roche), using the in situ cell death detection kit.

    Results: Canine plasmacytoma revealed moderate positive reaction with all methods. Canine carcinoma in situ showed moderate positive reaction with pro-ap-optotic antibodies and more intense with anti-apoptotic antibody. Dogs thymus showed moderate positive re-action in cortical thymocytes with pro-apoptotic anti-bodies and weak in the medullary thymocytes. With the anti-apoptotic antibody, medullary thymocytes

    showed weak reaction and negativity in the cortex. Identical results were obtained in rabbits thymus, but with stronger intensity.

    Conclusion: The rabbit thymus showed the most specific reaction of the tissues studied, by which the authors recommended it as a viable positive control for apoptosis studies.

    Acknowledgments: FCT and CI&DETS (PEst-OE/CED/UI4016)

  • SUPLEMENTO RPCV (2013) 108 (585-586) 69-91

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    Hrnia de Bexiga associada a Angioleio-miossarcoma Cutneo em Coelho

    Santos S.1*, Patrcio. R.1,2

    1 CICV - FMV- ULHT, Lisboa, Portugal2 AlcabidecheVet Clnica Veterinria, Cascais, Por-tugal

    * [email protected]

    Introduo: Em Agosto de 2012 um coelho de es-timao com 6 anos de idade, macho esterilizado foi submetido a cirurgia para extirpao de uma massa subcutnea ulcerada, localizada na regio inguinal e associada a uma hrnia de bexiga.

    Material e mtodos: Aps a cirurgia a massa foi enviada para anlise histopatolgica, fixando-se o te-cido em formol tamponado a 10% para colorao com hematoxilina & eosina (H&E). Realizou-se imunohis-toqumica com -actina (clone, 1A4 DAKO; 1\180)

    e desmina (clone DE-R-11, DAKO; 1\50) utilizando intestino e estmago de co como controlo positivo.

    Resultados: A H&E revelou uma neoplasia subcut-nea, no capsulada e infiltrativa constituda por clulas fusiformes anaplsicas organizadas em feixes compac-tos e ninhos. As clulas neoplsicas apresentavam-se intimamente relacionadas com as estruturas vasculares