Non-tuberculous Mycobacterial Granulomatous Dermatitis in an African Pygmy Hedgehog (Atelerix albiventris).

A 2-year-old female African pygmy hedgehog (Atelerix albiventris) with a cutaneous nodular lesion on the dorsal surface of the right forelimb was presented for clinical examination. Histopathological findings included granulomatous dermatitis with extensive necrosis. Long and slender acid-fast bacilli were seen within the cytoplasm of macrophages and in extracellular spaces. Bacteriological culture of skin revealed acid-fast bacilli and non-tuberculous mycobacterial infection was confirmed by gene sequencing and identity analysis using the BLAST tool. To our knowledge, this is the first report of non-tuberculous granulomatous dermatitis in hedgehogs.

Glial Response and Neuroinflammation in Cerebrocortical Atrophy in a Young Irish Wolfhound Dog.

A two-year-old, Irish Wolfhound dog presented with a history of progressive neurological signs. Neurological exam revealed disorientation, absence of menace response, reduction of right nasal sensation, hypermetria and ataxia with reduction of proprioception in all four limbs. MRI findings were compatible with laminar neuronal necrosis and possible bilateral cortical cerebral atrophy. Grossly, a severe bilateral reduction of the gray matter with flattening of gyri, mainly in frontal and parietal cerebral areas, was observed. Histologically, multiple, segmental, bilateral, and symmetric areas of neuronal loss, necrosis and degeneration, in a laminar pattern, associated with a reactive gliosis were observed. Immunohistochemical studies showed severe reduction of neuronal bodies, proliferation and hypertrophy of astrocytes and microglia. Few perivascular B and T cells were demonstrated. Based on these data, we show some of the neuroinflammatory events that occur during CNS repair in a chronic phase of this condition.

Poxviruses diagnosed in cattle from Distrito Federal, Brazil (2015-2018).

A retrospective study of officially diagnosed poxvirus infections in cattle in Distrito Federal (DF), Brazil, between 2015 and 2018 was performed. All cases were investigated by the DF Official Veterinary Service. In the most cases, samples of oral, cutaneous (teats, udder) or foot lesions were submitted to molecular diagnosis by PCR. In approximately 70% of the cases, additional samples were also submitted for histopathology. Ninety-three out of 2,467 clinically examined cattle (from 385 farms) presented suggestive and/or compatible lesions with poxviruses. Fifty-two out of these 93 cases were confirmed as poxviruses: 27 vaccinia virus (VACV), 9 pseudocowpox virus (PCPV), 8 bovine papular stomatitis virus (BPSV), 5 coinfection by PCPV and BPSV and 3 unidentified parapoxvirus. The clinical cases were observed in farms with different exploration (beef, dairy or mixed) from 9 out of 30 administrative regions of DF. Gross findings consisted of papules, vesicles, ulcers, scabs and scars and varied of type, severity and affected tissue, according to the detected virus. A single human case was observed associated with a BPSV infection. Histologically, the lesions were very similar, independently of the detected poxvirus, and included mild to moderate, superficial, multifocal inflammatory infiltrate of lymphocytes, plasma cells, macrophages and/or neutrophils, with acanthosis and parakeratotic hyperkeratosis, usually associated with serous content, cellular debris and spongiosis. In the ulcerated lesions, there were focally extensive areas of necrosis with severe infiltrate of neutrophils in the adjacent connective tissue. Few to moderate amount of 4- to 8-µm eosinophilic inclusion bodies were observed in the cytoplasm of keratinocytes in 6 cases (2 of VACV, 2 of PCPV and 2 of PCPV/BPSV coinfection). Data of the current study demonstrate the wide circulation of different poxviruses in cattle from DF.

Evidence of bovine viral diarrhea virus transmission by back pond water in experimentally infected piglets / Evidência da transmissão do vírus da diarreia viral bovina através da lâmina d'água em leitões experimentalmente infectados

Swine can be infected by bovine viral diarrhea virus (BVDV). However, transmission routes among pigs are still unknown. The objective of the present study was to induce experimental infection of BVDV-1 in weaned piglets and to assess the potential transmission through pen back pond water, used to facilitate heat exchange of the pigs housed in barns. Two repetitions (BP1 and BP 2) were performed using 12 piglets proven to be free BVDV (n=6 per repetition) allocated into three groups: control, sentinels and infected with two piglets each. The piglets were placed in stainless steel isolators. The infected group received an inoculum containing BVDV-1, Singer strain. The piglets remained in the cabinets for 25 days, during which samples of nasal swab were collected daily and blood sampled weekly. At the end, the piglets were euthanized, necropsied and organ fragments were collected for histopathology, immunohistochemistry and RT-PCR. In the first experiment (BP1) the infected animals shed the virus between days 6 and 21 post-infection. Regarding the sentinel group, shedding occurred in only one piglet, on the 20th day after infection, and seroconversion was observed on the 25th day post-infection. In BP2, infected piglets I3 and I4 shed the virus on days 4 and 21 post-infection, respectively. Only one sentinel piglet (S3) she the virus on day 13 post-infection. Therefore, it was concluded that pigs can become infected with BVDV-1 and shed potentially infectious viral particles consequently, being able to transmit the virus to other pigs through back pond water.(AU)

Os suínos podem ser infectados pelo vírus da diarreia viral bovina (BVDV). No entanto, as vias de transmissão entre os suínos são ainda desconhecidas. O objetivo do presente estudo foi induzir a infecção experimental de BVDV-1 em leitões desmamados e avaliar a potencial transmissão pela lâmina d'água, que ajuda na troca de calor dos suínos alojados em baias. Duas repetições do experimento (BP1 e BP2) foram realizadas com 12 animais comprovadamente livres de BVDV (n=6 por repetição) alocados em três grupos: controle, sentinelas e infectados, com dois animais cada. Os animais foram mantidos em isoladores de aço inoxidável. O grupo infectado recebeu um inóculo contendo BVDV-1, estirpe Singer. Os animais permaneceram nos isoladores durante 25 dias e, durante esse período, amostras de suabe nasal foram coletadas diariamente e sangue coletado semanalmente. No final, os animais foram eutanasiados, necropsiados e fragmentos de órgãos foram coletados para histopatologia, imuno-histoquímica e RT-PCR. No primeiro experimento (BP1), os animais infectados excretaram partículas virais entre os dias 6 e 21 pós-infecção. Quanto ao grupo sentinela, a excreção ocorreu apenas em um animal, no 20º dia pós-infecção, e a soroconversão foi observada no 25º dia pós-infecção. Na BP2, os animais infectados I3 e I4 excretaram partículas virais nos dias 4 e 21 pós-infecção, respectivamente. Apenas um animal sentinela (S3) apresentou excreção no dia 13 pós-infecção. Concluiu-se que os suínos podem se infectar com BVDV-1 e excretar partículas virais potencialmente infecciosas, sendo capazes de transmitir o vírus a outros suínos através da lâmina d'água.(AU)

Neurological diseases of cattle in the state of Goiás, Brazil (2010-2017) / Doenças neurológicas de bovinos no estado de Goiás (2010-2017)

A retrospective study was conducted on neurological diseases of cattle in the state of Goiás, Brazil, from March 2010 to August 2017. Samples of three veterinary diagnostic laboratories were analyzed. Diagnosis was established in 170 out of 407 cattle with neurological signs. Epidemiological, clinical, and anatomic pathology features of each case were researched in the files. Main disorders included diseases caused by viruses (rabies 29.41%, meningoencephalitis by bovine herpesvirus 15.88%, and malignant catarrhal fever 1.76%), by bacteria (botulism 5.88%, suppurative meningitis 3.53%, encephalic abscesses 2.94%, listeriosis 1.76%, and thrombotic meningoencephalitis 1.76%), of metabolic origin (polioencephalomalacia 17.06%), of indefinite cause (lymphoplasmacytic meningoencephalitis 11.18%, traumatic hemorrhages 3.53%, and multifocal malacia with gliosis 1.18%), congenital (hydrocephaly 1.18% and multiple malformations 0.59%), toxic (urea poisoning 1.18% and insecticide poisoning 0.59%), and parasitic (meningoencephalitis associated with infection by Trypanosoma sp. 0.59%).(AU)

Foi realizado um estudo retrospectivo de doenças neurológicas de bovinos no estado de Goiás durante o período de março de 2010 a agosto de 2017, analisando amostras de três laboratórios de diagnóstico veterinário. De 407 bovinos que apresentaram sinais clínicos neurológicos, o diagnóstico foi estabelecido em 170 casos. Desses casos, foram pesquisadas nas fichas as características epidemiológicas, clínicas e anatomopatológicas. As principais doenças diagnosticadas foram causadas por vírus (raiva 29,41%, meningoencefalite por herpesvírus bovino 15,88% e febre catarral maligna 1,76%), de origem metabólica (polioencefalomalacia 17,06%), por bactérias (botulismo 5,88%, meningite supurativa 3,53%, abscessos encefálicos 2,94%, listeriose 1,76% e meningoencefalite trombótica 1,76%), sem causa definida (meningoencefalite linfoplasmocítica 11,18%, hemorragias traumáticas 3,53% e malacia multifocal com gliose 1,18%), congênitas (hidrocefalia 1,18% e malformações múltiplas 0,59%), tóxicas (intoxicação por ureia 1,18% e intoxicação por inseticida 0,59%), e parasitária (meningoencefalite associada à infecção por Trypanosoma sp. 0,59%).(AU)

Alterações neoplásicas e não neoplásicas das mãos e dos pés de cães (2003-2016) / Neoplastic and non-neoplastic lesions of the hands and feet of dogs (2003-2016)

Mãos e pés de cães são comumente afetados por lesões neoplásicas e não neoplásicas. Estas alterações podem apresentar prognóstico histopatológico ou clínico ruim e a amputação tende a ser o tratamento de escolha. Estudos prévios avaliando a prevalência e os aspectos clínicopatológicos de alterações digitais em cães têm sido realizados em outros países, entretanto trabalhos similares não foram realizados no Brasil. O objetivo do presente estudo foi descrever e caracterizar histologicamente as lesões das mãos e pés de cães. Amostras de 105 lesões tumoriformes das mãos e pés de cães foram avaliadas. Essas amostras foram coletadas entre 2003 e 2016 e foram obtidas de três laboratórios brasileiros de diagnóstico veterinário. Todos espécimes foram fixados em formol tamponado 10%, processados rotineiramente e corados por hematoxilina-eosina. Adicionalmente, as colorações de azul de toluidina, Brown e Breen, periodic acid Schiff (PAS), Grocott Methenamine Silver (GMS) e Ziehl Neelsen foram utilizados em casos específicos. Imuno-histoquímica foi realizada usando os anticorpos anti-Leishmania spp., anti-CD117, anti-CD79, anti-CD3, anti-Melan A, anti-lisozima, anti-Pancytokeratin AE1/AE3 e anti-vimentina. Os neoplasmas (62,9%) foram as alterações mais frequentes, seguidas por inflamações (19%) e outras alterações (18,1%). Entre os neoplasmas, 40,9% foram mesenquimais, 39,4% epiteliais e 19,7% de células redondas. Todos os neoplasmas de células redondas e a maioria dos epiteliais e mesenquimais eram malignos. Não se observou predileção de prevalência dos neoplasmas em relação ao sexo. As raças Labrador Retriever, Schnauzer, Teckel, SRD, Pastor Alemão, Rottweiler e Pit Bull foram as mais afetadas. Fila Brasileiro, Pit Bull e Schnauzer tiveram alta incidência de neoplasmas mesenquimais, epiteliais e de células redondas, respectivamente. Inflamação foi mais comumente observada em cães Labrador Retriever e as outras alterações em Teckel, Labrador Retriever e SRD. A idade e o peso médio dos animais afetados foram de 8,4 anos e 28,5 kg, respectivamente. O diâmetro médio das lesões tumoriformes foi de 2,5 cm e as lesões neoplásicas apresentaram as maiores médias. As lesões ocorreram principalmente em animais de pelagem amarela. A maioria das biópsias incisionais (56,4%) e amputações (85,3%) consistiram de neoplasmas. O principal membro afetado foi o torácico direito e o dígito foi a estrutura anatômica acometida mais frequentemente. Carcinoma de células escamosas (14,2%) foi o neoplasma mais frequente, seguido do mastocitoma (7,6%), melanoma (7,6%) e sarcoma indiferenciado (7,6%). Em metade dos casos de inflamação, a lesão acometeu o folículo piloso e derme adjacente, e o infiltrado foi predominantemente piogranulomatoso ou lnfoplasmocítico. Cistos foliculares, calcinose circunscrita e acrocordoma foram as principais lesões não neoplásicas e não inflamatórias diagnosticadas.(AU)

Hands and feet of dogs are commonly affected by neoplastic and non-neoplastic lesions. These lesions may commonly present poor clinical or histopathological prognosis and amputation tends to be the treatment of choice. Previous studies regarding the prevalence and the clinicopathological aspects of digital changes in dogs have been performed abroad, with no similar investigations in Brazil. The aim of this manuscript was to perform a histopathological study to characterize hands and feet lesions of dogs from Brazil.Tissue samples from 105 tumor-like lesions of dogs hands and feet were used in this study. The samples came from three Brazilian veterinary diagnostic laboratories and were collected between 2003 and 2016. All specimens were fixed in 10% buffered formalin, and routinely processed for paraffin embedding and hematoxylin-eosin staining. Additionally, toluidine blue, Brown and Breen, periodic acid Schiff (PAS), Grocott Methenamine Silver (GMS) and Ziehl Neelsen were used in selected cases. Immunohistochemistry using anti-Leishmania spp., anti-CD117, anti-CD79, anti-CD3, anti-Melan A, anti-lysozyme, anti-Pancytokeratin AE1/AE3, and anti-vimentin antibodies were also performed. Neoplasia (62.9%) was the most frequently found lesion, followed by inflammation (19%), and other changes (18.1%). Among the neoplasms, 40.9% were mesenchymal, 39.4% epithelial, and 19.7% round cells. All round cells and most of the epithelial and mesenchymal neoplasms were malignant. There was no difference between males and females regarding neoplasia prevalence. Labrador Retriever, Schnauzer, Teckel, mongrel, German Shepherd, Rottweiler, and Pit Bull were the most frequently affected breeds. Fila Brasileiro (aka Brazilian Mastiff), Pit Bull and Schnauzer had higher incidence of mesenchymal, epithelial, and round cells neoplasms, respectively. Inflammation was more commonly observed in the Labrador Retriever and other changes were more commonly in Teckel, Labrador Retriever, and mongrel dogs. Medium age and weight were 8.4 years and 28.5 kg, respectively. Medium diameter of the tumor-like lesions was 2.5 cm and the neoplastic lesions presented higher averages. Lesions were mainly in yellow-coated dogs. Incisional biopsy samples (56.4%) and amputations (85.3%) were mostly from neoplastic cases. Right forelimb was the most affected limb, and digits were the main injured anatomic structures. Squamous cell carcinoma was the most (14.2%) commonly diagnosed neoplasm, followed by mast cell tumor (7.6%), melanoma (7.6%), and undifferentiated sarcoma (7.6%). In half of the inflammatory cases, the lesion was centered in hair follicle and surrounding dermis. The inflammatory infiltrate was predominantly piogranulomatous or lymphoplasmacytic. Follicular cyst, calcinosis circumscripta, and acrochordons were the main non neoplastic and non-inflammatory changes.(AU)

Caracterização etiológica, epidemiológica e clínico-patológica da meningoencefalite por herpesvírus bovino em bovinos no Estado de Goiás / Etiological, epidemiological, clinical and pathological characterization of meningoencephalitis in cattle by bovine herpesvirus in the State of Goiás, Brazil

Foram estudados 26 casos de meningoencefalite por herpesvírus bovino (BoHV) diagnosticados entre 2010-2016, no Estado de Goiás (GO). A doença acometeu principalmente bovinos jovens, entre 60 dias a 18 meses de idade. Não houve associação entre os casos e o sexo dos bovinos e a sazonalidade. A doença foi observada em todas as cinco Mesorregiões do Estado, com uma frequência maior nas Mesorregiões Sul e Centro. Os sinais clínicos mais frequentemente observados incluíram cegueira, incoordenação, sialorreia e ataxia. As principais alterações macroscópicas observadas incluíram congestão com tumefação e achatamento das circunvoluções, amolecimento e amarelamento do córtex telencefálico e focos de hemorragia. Em cinco encéfalos, não foram observadas alterações macroscópicas e em quatro as alterações não foram informadas. As principais alterações histológicas ocorreram no córtex telencefálico, principalmente o córtex frontal e parietal, mas em alguns casos, lesões de menor intensidade foram também observadas no tálamo, núcleos basais, mesencéfalo, ponte, bulbo, cerebelo e hipocampo. Todos os casos apresentaram meningoencefalite linfoplasmocítica e corpúsculos de inclusão intranucleares basofílicos em astrócitos e, eventualmente, em neurônios. Outras lesões frequentes incluíram necrose neuronal laminar segmentar (neurônio vermelho), espongiose, tumefação do núcleo das células endoteliais, gliose focal ou difusa, hipertrofia de astrócitos, infiltração por células gitter, congestão e hemorragia. Lesões menos comuns incluíram astrócitos Alzheimer tipo II, lesão residual e neuronofagia. A necrose neuronal e o edema (espongiose) foram mais acentuados nas camadas granular externa, molecular, de células piramidais e granular interna dos telencéfalos. Tanto os giros quanto os sulcos foram afetados igualmente. Dos 26 casos, o DNA de BoHV-5 foi amplificado em dois (7,69%) casos, enquanto que o de BoHV-1 foi identificado em um caso (3,84%). Nos casos positivos para BoHV-5 foram usadas amostras fixadas em formol a 10% e incluídas em parafina e amostras congeladas foram utilizadas no caso positivo para BoHV-1.(AU)

Twenty six cases of bovine herpetic meningoencephalitis diagnosed from 2010-2016 in Goiás state, Brazil, were studied. Affected cattle were mainly 60-day to 18-month-old. There was no association of the disease with sex and seasonality. The disease was found in all five mesoregions with a higher prevalence in southern and central state of Goiás. Clinical signs more frequently observed included blindness, incoordination, circling, excessive salivation, and ataxia. Main gross findings in the brain were congestion with swelling and flattening of gyri, softening and yellow discoloration of cerebral cortex and hemorrhagic foci. In five cases no gross changes were observed in the brain and in four cases there is no information. The main histopathological changes were in the cortex of telencephalic lobes, especially the frontal and parietal; however less prominent and less frequently found lesions occurred in the thalamus, basal nuclei, midbrain, pons, medulla oblongata, cerebellum, and hippocampus. All cases presented lymphoplasmocytic meningoencephalitis and intranuclear basophilic inclusion bodies in astrocytes, less commonly in neurons. Other frequent lesions included segmental laminar neuronal necrosis (red neurons), spongiosis, swollen vascular endothelial nuclei, gliosis (focal and diffuse), hypertrophy of astrocytes, infiltration of gitter cells, congestion, and hemorrhage. Lesions less frequently observed were Alzheimer type II astrocytes, residual lesion and neuronophagia. The most frequently affected cortical layers by neuronal necrosis and edema were external and internal granular, molecular, and pyramidal cell layers. Gyri and sulci were equally affected. Of the 26 cases, in 2 (7.69%) the DNA of BoHV-5 was amplified with samples fixed in 10% formalin and paraffin-embedded. DNA of BoHV-1 was identified in another case (3.84%) where, positive to BoHV-1, fresh samples were used.(AU)

Caracterização clínica e histopatológica de bulbos oculares de cães e gatos (2005-2015) / Clinical and histopathologic characterization of ocular bulbs in dogs and cats (2005-2015)

Objetivou-se realizar a caracterização clínica e histopatológica de bulbos oculares de cães e gatos, removidos cirurgicamente por indicação clínica, no período entre 2005-2015. Foram realizados 101 procedimentos de remoção do bulbo ocular, 93 enucleações (92%) e 8 exenterações (8%). Os procedimentos foram realizados em 80 cães, (79% dos casos) e em 21 gatos (21% dos casos). Os cães submetidos à intervenção cirúrgica apresentavam perfurações oculares (n=31, 39%), glaucoma (n=19, 24%), protrusão ocular (n=13, 16%), diagnósticos sugestivos de neoplasia (n=10, 12,5%) e outros (n=3, 2,97%). Dentre os gatos os diagnósticos clínicos compreenderam perfuração ocular (n=9, 49%), glaucoma (n=3, 14,2%), microftalmia (n=3, 14,2%), sugestivo de neoplasia (n=2, 9,5%), protrusão ocular (n=2, 9,5%) e outros (n=2, 9,5%). Vinte e sete amostras de bulbos oculares foram submetidas para avaliação histopatológica, sendo 23 provenientes de cães e quatro de gatos. Dentre as amostras de origem canina, sete exibiram achados histopatológicos compatíveis com glaucoma, sete perfurações oculares, seis neoplasias intraoculares, duas panoftalmites e um quadro de Phthisis bulbi. Nos bulbos oculares de gatos foram diagnosticadas duas neoplasias intraoculares, uma microftalmia e uma perfuração ocular. Constatou-se que o glaucoma secundário, as neoplasias e as perfurações oculares estão entre as principais causas de enucleação em cães, e associadas a alterações primárias como a ceratoconjuntivite seca e as uveítes. Estas afecções, diferentemente das neoplasias, podem ter êxito terapêutico mediante diagnóstico e tratamento precoces, prevenindo a cegueira e a remoção do bulbo ocular.(AU)

This study aimed to carry out the clinical and histopathologic characterization of ocular bulbs from dogs and cats, surgical removed as clinical indication between 2005-2015. Hundred-one procedures were performed, 93 enucleations (92%) and 8 exenterantions (8%). The procedures were performed on 80 dogs (79% of cases) and 21 cats (21% of cases). The dogs underwent to surgery had ocular perfuration (n=31, 39%), glaucoma (n=19, 24%), ocular protrusion (n=13, 16%), diagnosis suggestive of neoplasia (n=10, 12, 5%) and other (n=3, 2.97%). Among the clinical diagnosis cats comprise ocular perforation (n=9, 49%), glaucoma (n=3, 14.2%), microphthalmia (n=3, 14.2%), suggestive of neoplasia (n=2, 9.5%), ocular protrusion (n=2, 9.5%) and other (n=2, 9.5%). Twenty-seven samples of ocular bulbs were sent for histopathologic evaluation in Veterinary Pathology Laboratory, being 23 from dogs and four cats. Among the samples of dogs, seven exhibited histopathologic findings compatible with glaucoma, seven ocular perforations, six had intraocular neoplasias, two panoftalmites and condition of Phthisis bulbi. The ocular bulbs of cats received the diagnosis of two intraocular neoplasias, one microphthalmia and ocular perforation. It was found that the secondary glaucoma and ocular perforations are among the leading causes of enucleation in dogs, and associated with primary disorders such as keratoconjunctivitis sicca and uveitis. These disorders, unlike neoplasia, may have therapeutic success through early diagnosis and treatment, preventing blindness and removal of the eyeball.(AU)

Cytoskeletal proteins in the follicular wall of normal andcystic ovaries of sows

The expression of cytoskeletal proteins was evaluated immunohistochemically in 36 normal ovaries sampled from 18 sows and 44 cystic ovaries sampled from of 22 sows, was evaluated. All sows had history of reproductive problems, such as infertility or subfertility. The immunohistochemically stained area (IHCSA) was quantified through image analysis to evaluate the expression of these proteins in the follicular wall of secondary, tertiary, and cystic follicles. Cytokeratins (CK) immunoreactivity was strong in the granulosa cell layer (GC) and mild in the theca interna (TI) and externa (TE) of the normal follicles. There was severe reduction of the reaction to CK in the GC in the cystic follicles, mainly in the luteinized cysts. The immunoreactivity for vimentin was higher in the GC from normal and cystic follicles in contrast with the other follicular structures. In the luteinized cysts, the IHCSA for vimentin was significantly higher in TI and in both observed cysts, the labeling was more accentuated in TE. Immunohistochemical detection of desmin and -SMA was restricted to the TE, without differences between the normal and cystic follicles. The results of the current study show that the development of ovarian cysts in sows is associated to changes in the expression of the cytoskeletal proteins CK and vimentin.

A expressão de proteínas do citoesqueleto foi avaliada por imuno-histoquímica em ovários normais e císticos de porcas matrizes. Amostras de 36 ovários normais (18 porcas) e de 44 císticos (22 porcas) foram avaliadas. Todas as matrizes apresentaram histórico de problemas reprodutivos, como infertilidade ou subfertilidade. As áreas coradas por imuno-histoquímica (IHCSA) foram quantificadas por avaliação de imagens avaliando a expressão dessas proteínas na parede folicular de folículos secundários, terciários e císticos. A imuno-reatividade para citoqueratina (CK) foi forte na camada de células da granulosa (GC) e discreta nas tecas interna (TI) e externa (TE) dos folículos normais. Houve redução acentuada da reação de CK na CG dos folículos císticos, principalmente nos cistos luteinizados. A reação para vimentina foi mais intensa na CG dos folículos normais e císticos em comparação com outras estruturas foliculares. Nos cistos luteinizados, a IHCSA para vimentina foi significativamente maior na TI e, em ambos os cistos observados, a marcação foi mais acentuada na TE. A marcação de desmina e actina alfa de músculo liso foi restrita a TE, sem diferenças entre os folículos normais e císticos. Os resultados do presente estudo mostram que o desenvolvimento de cistos ovarianos em porcas matrizes está associado a alterações na expressão das proteínas do citoesqueleto CK e vimentina.

Aspectos anatomopatológicos do parasitismo por nematódeos da superfamília filarioidea em capivaras (hydrochoerus hydrochaeris) de vida livre no Centro-Oeste brasileiro / Anatomic pathology aspects of the parasitism by cruorifilaria tuberocauda in capybaras (hydrochoerus hydrochaeris) in midwestern Brazil

São descritos dois casos de parasitismo por Cruorifilaria tuberocauda em capivaras de vida livre no Distrito Federal, Brasil. Macroscopicamente, observou-se nas superfícies de corte dos rins espessamento acentuado de vasos das regiões cortical e córtico-medular. Microscopicamente, havia arterite proliferativa e granulomatosa acentuada associada a filarídeos intralesionais consistentes com Cruorifilaria tuberocauda. Esse é o primeiro relato do parasitismo por esse filarídeo em capivaras no Distrito Federal.

This report describes two cases of parasitism by Cruorifilaria tuberocauda in wild capybaras (Hydrochoerus hydrochaeris) from Distrito Federal, Brazil. Grossly, there was marked thickening of vessels wall of the cortical and corticomedullary regions of both kidneys. Microscopically, there was severe proliferative and granulomatous arteritis associated with intralesional filarids, consistent with Cruorifilaria tuberocauda. For the first time this filarid is reported parasitizing capybaras in Distrito Federal, Brazil.

Spirocerca lupi granulomatous pneumonia in two free-ranging maned wolves (Chrysocyon brachyurus) from central Brazil.

The current report describes granulomatous pneumonia due to Spirocerca lupi in 2 free-ranging maned wolves (Chrysocyon brachyurus). Both wolves had multiple, white, 1-1.5 cm in diameter, soft, encapsulated granulomas in the caudal lung lobes, which contained centrally placed parasites on cut sections. Microscopically, there was granulomatous inflammation with numerous intralesional sections of spirurid nematodes. Representative complete adult specimens of nematodes derived from these lesions were submitted for parasitological exam and identified as the spirurid S. lupi. To the authors' knowledge, there have been no published reports of S. lupi in maned wolves.

Mammary diffuse fibroadenomatoid hyperplasia in water buffalo (Bubalus bubalis): three cases.

The current report describes 3 rare cases of mammary diffuse fibroadenomatoid hyperplasia in water buffalo (Bubalus bubalis). All of the animals were between 10 and 12 months of age. Grossly, the lesions consisted of severe diffuse swelling with homogeneous large masses in the udder. Surgical removal of the masses was curative. Microscopically, there was severe hyperplasia of the mammary epithelium and numerous well-differentiated and mildly pleomorphic acini and their associated ducts. Moderate proliferation of the fibrous connective tissue and the myoepithelial cells near the proliferating acini was also evident. The hyperplastic epithelial cells exhibited positive immunostaining for cytokeratin, estrogen receptors, and progesterone receptors. In addition, the myoepithelial cells displayed moderate positivity for alpha smooth muscle actin. Based on the clinical, morphologic, and immunohistochemical findings, a diagnosis of mammary diffuse fibroadenomatoid hyperplasia with probable hormonal influence was made.

Outbreaks of vesicular disease caused by Vaccinia virus in dairy cattle from Goiás State, Brazil (2010-2012) / Surtos de doença vesicular causados pelo Vaccinia virus em bovinos de leite no Estado de Goiás, Brasil (2010-2012)

Cases of vesicular and exanthematic disease by Vaccinia virus (VACV) have been reported in dairy herds of several Brazilian regions, occasionally also affecting humans. The present article describes eight outbreaks of vesicular disease caused by VACV in dairy herds of six counties of Goiás state, Midwestern Brazil (2010-2012), involving a total of 122 cows, 12 calves and 11 people. Dairy cows (3 to 9 years old) were affected in all cases and calves (2 to 9 months old) were affected in five outbreaks, presenting oral lesions. The morbidity ranged between 8 and 100% in cows, and 1.5 to 31% in calves. In the cows, the clinical signs started with vesicles (2-7mm), painful and coalescent papules (3-8 mm), which resulted in ulcers (5-25mm) and scabs in teats, and, occasionally, in the muzzle. The clinical course lasted from 16 to 26 days. The histopathology of bovine skin samples revealed superficial perivascular inflammatory infiltrate of lymphocytes, plasma cells, neutrophils, macrophages and multifocal areas of acanthosis, spongiosis, hipergranulosis and parakeratotic or orthokeratotic hyperkeratosis with adjacent focally extensive ulcers. Eosinophilic inclusion bodies were noted in the cytoplasm of the keratinocytes. PCR to vgf gene of Orthopoxvirus was positive in samples collected from all outbreaks, and in some cases, genomic VACV sequences were identified by nucleotide sequencing of the PCR amplicons. Infectious virus was isolated in cell culture from scabs from one outbreak. Antibodies to Orthopoxvirus were detected in at least 3 or 4 animals in most outbreaks, by ELISA (outbreaks 1, 2, 3, 4, 5 and 7) or virus-neutralization (outbreak 6). Neutralizing titers ranging from 8 to 64 in outbreak 6. In all outbreaks, VACV infection was suspected based on the clinical and pathological findings and it was confirmed by laboratory tests. Upon the etiological confirmation, other agents associated with vesicular disease were discarded. In all outbreaks, at least one milker who handled the affected cows developed malaise, headache, fever, painful vesico-pustular lesions mainly in the hands, but also in the neck and nose. These results confirm the circulation of VACV in the region and call attention for a correct diagnosis and the adoption of prophylactic and control measures.

Casos de doença vesicular e exantemática associados ao Vaccinia virus (VACV) têm sido descritos em rebanhos leiteiros em diversas regiões do Brasil, ocasionalmente afetando também os ordenhadores. Este artigo descreve oito surtos de doença vesicular associados ao VACV ocorridos em rebanhos leiteiros de seis municípios do estado de Goiás (2010-2012), com o envolvimento de 122 vacas em lactação, de 12 bezerros e de 11 pessoas. Vacas em lactação (3-9 anos de idade) foram afetadas em todos os casos. Em cinco rebanhos, bezerros de 2-9 meses apresentaram lesões orais. A morbidade nos rebanhos variou entre oito e 100% (vacas) e entre 1,5 e 31% (bezerros). As lesões iniciavam como vesículas (2-7mm) ou pápulas doloridas e coalescentes (3-8mm), que progrediam para úlceras (5-25mm) e crostas, localizadas principalmente nas tetas e, eventualmente, no focinho das vacas. O curso clínico variou entre 16 e 26 dias. Histopatologia de amostras de pele coletadas de bovinos revelou dermatite perivascular superficial com infiltrado de linfócitos, neutrófilos, plasmócitos e macrófagos, além de áreas multifocais de acantose, espongiose, hipergranulose e hiperceratose ortoceratótica ou paraceratótica com úlceras focalmente extensas. No citoplasma dos ceratinócitos adjacentes às úlceras, observaram-se numerosos corpúsculos de inclusão eosinofílicos. Em todos os surtos, amostras de lesões cutâneas dos bovinos foram positivas para o gene vgf dos Orthopoxvirus por PCR, e em alguns casos, a identificação do VACV foi confirmada por sequenciamento de nucleotídeos dos amplicons. O vírus foi detectado por isolamento em cultivo celular em um dos surtos e, pelo menos 2 a 3 vacas por rebanho, apresentaram sorologia positiva para Orthopoxvirus pelos testes de ELISA (surtos 1, 2, 3, 4, 5 e 7) e soroneutralização (surto 6). No surto 6, os títulos de anticorpos neutralizantes variaram de 8 a 64. O diagnóstico da infecção pelo VACV, inicialmente suspeito com base nos achados clínicos e patológicos, foi confirmado em todos os surtos por exames laboratoriais. Em todos os surtos, pelo menos um ordenhador que teve contato com os bovinos afetados apresentou mal-estar geral, febre alta, dor de cabeça e lesões vesiculo-pustulosas doloridas, principalmente nas mãos, mas também no pescoço e nariz. Esses resultados confirmam a circulação do VACV no rebanho bovino da região centro-oeste, alertando para necessidade de diagnóstico correto e adoção de medidas profiláticas e de controle.

Coinfection by Vaccinia virus and an Orf virus-like parapoxvirus in an outbreak of vesicular disease in dairy cows in midwestern Brazil.

The current report describes an outbreak of vesicular disease affecting dairy cows in midwestern Brazil in which a coinfection with 2 poxviruses-Vaccinia virus (VACV) and a parapoxvirus-was demonstrated. Milking cows presented vesicles, painful reddish or whitish papules, and scabby proliferative lesions in the teats and udder, in a clinical course of approximately 10-21 days. Histologically, multifocal areas of moderate to severe acanthosis, spongiosis, hypergranulosis, and parakeratotic or orthokeratotic hyperkeratosis with adjacent focally extensive ulcers were observed in the epidermis. Rounded eosinophilic inclusion bodies were observed in the cytoplasm of epithelial cells of areas with acanthosis or necrosis. Moderate inflammatory infiltrate of lymphocytes, plasma cells, neutrophils, and macrophages were observed in some dermal areas. Two people milking the affected cows developed lesions on the hands, painful papules which progressed to ulcerative and scabby lesions in 4-7 days. Electron microscopy of scabs from 1 cow revealed the concomitant presence of orthopoxvirus and parapoxvirus particles. Scabs from 2 cows were positive by polymerase chain reaction for the parapoxvirus B2L gene; 1 of the scabs was also positive for the VACV vgf gene. Nucleotide sequencing of the B2L amplicon revealed a similarity of 96-99% with Orf virus (ORFV) and lower identity with Pseudocowpox virus (92-95%) and Bovine papular stomatitis virus (85-86%). Nucleotide sequencing of a region of parapoxvirus DNA polymerase gene revealed a high similarity (98-100%) with ORFV sequences. Thus, an unusual coinfection with VACV and a parapoxvirus, likely ORFV, was demonstrated in the outbreak.

Intoxicação espontânea por Pterodon emarginatus (Fabaceae) em bovinos no estado de Goiás / Spontaneous poisoning of cattle by Pterodon emarginatus (Fabaceae) in Goiás, Brazil

Descreve-se um surto de intoxicação por Pterodon emarginatus em bovinos no Estado de Goiás. De um lote de 84 bovinos que comeram avidamente folhas e os frutos da planta após a queda acidental de uma árvore, todos os animais adoeceram e sete morreram. Os sinais clínicos observados foram eriçamento dos pelos, retração do flanco, apatia profunda, prostração, tremores musculares, ressecamento do focinho, tenesmo, incoordenação, relutância em movimentar-se e decúbito esternal prolongado. As atividades séricas de AST, ALT e GGT e os teores de bilirrubina estavam acentuadamente elevados. As principais alterações macroscópicas consistiam de hepatomegalia e áreas multifocais de necrose no fígado, além de hemorragias no coração, pleura parietal, mesentério, omento, serosa do rúmen, baço, pulmão, subcutâneo e musculatura esquelética intercostal e torácica. Microscopicamente, observou-se degeneração e necrose hepatocelular massiva moderada a acentuada, hiperplasia biliar multifocal moderada e bilestase multifocal leve. Adicionalmente, notou-se degeneração vacuolar multifocal moderada nos túbulos contorcidos dos rins.

An outbreak of poisoning by Pterodon emarginatus in cattle in Goiás, Brazil, is described. Eighty four cattle that accidentally consumed the leaves and fruits from a P. emarginatus tree, fallen in a storm, were affected and seven died. Clinical signs included piloerection, flank retraction, marked apathy, prostration, muscle tremors, muzzle dryness, tenesmus, incoordination, reluctance to move, and prolonged sternal recumbency. AST, ALT, and GGT serum activity, and bilirubin levels were markedly increased. Gross lesions included hepatomegaly, multifocal areas of necrosis in the liver, and hemorrhages in the heart, parietal pleura, mesentery, omentum, ruminal serosa, spleen, lung, subcutaneous tissue, and thoracic and intercostal skeletal muscles. Histologically, moderate to severe, massive hepatocellular degeneration and necrosis with moderate bile duct hyperplasia and mild bile stasis were observed. In addition, moderate multifocal vacuolar degeneration in the convoluted renal tubular cells was noted.

Bovine papular stomatitis affecting dairy cows and milkers in midwestern Brazil.

Bovine papular stomatitis virus (BPSV) is a parapoxvirus associated with papular and erosive lesions on the muzzle, lips, and oral mucosa of cattle. Teats of milking cows are occasionally affected, and the infection is frequently transmitted to human beings. The present report describes an outbreak of BPSV infection affecting cows in midwestern Brazil, with human involvement. The disease was observed in neighboring small hand-milking farms, affecting 20 milking cows. The signs included painful reddish papules, ulcers, and scabby proliferative lesions on the teats, with a clinical course of 7-12 days. Affected cows presented severe local pain, not allowing the completion of milking. Histologically, acanthosis, spongiosis, and parakeratotic hyperkeratosis with adjacent focally extensive ulcers and multifocal inflammatory infiltrate were observed in the epidermis. Eosinophilic inclusion bodies were noted in the cytoplasm of epithelial cells. Personnel milking the affected cows developed lesions on the hands, painful papules that progressed to ulcerative and scabby lesions in 4-7 days. A polymerase chain reaction using a set of pan-parapoxvirus primers for the B2L gene performed on DNA extracted from scabs amplified a 590-bp product, which when sequenced, revealed similarities of 99%, 85%, and 84% with BPSV, Pseudocowpox virus, and Orf virus, respectively. A phylogenetic tree based on the B2L sequence was constructed, showing that the virus clustered with BPSV isolates. Although clinical cases compatible with BSPV infection have been frequently described in Brazil, the present report identifies the agent associated with cattle and human disease in the country.

Surtos de infecção por Piscinoodinium pillulare e Henneguya spp. em pacus (Piaractus mesopotamicus) criados intensivamente no Sudoeste de Goiás / Outbreaks of Piscinoodinium pillulare and Henneguya spp. infection in intensively raised Piaractus mesopotamicus in Southwestern Goiás, Brazil

Três surtos de infecção por Piscinoodinium pillulare e Henneguya spp. em Piaractus mesopotamicus criados intensivamente no Sudoeste de Goiás são descritos. Dois surtos ocorreram no inverno e um no verão. As taxas de morbidade (58,57%-90%) e mortalidade (100%) foram determinadas. Episódios de estresse ocorreram previamente em todos os casos. Sinais clínicos incluíam mudanças comportamentais e movimentos erráticos com perda de equilíbrio. Os principais achados macroscópicos foram tumefação das brânquias com exsudato mucoso esbranquiçado. Microscopicamente, as brânquias apresentaram inflamação linfoplasmocítica e necrosante, multifocal, leve a moderada, com hipertrofia e fusão de lamelas secundárias. Adicionalmente, numerosos trofontes intralesionais de P. pillulare e cistos de Henneguya spp. foram notados. Para o conhecimento dos autores, essa parece ser a primeira descrição de infecção por P. pillulare e Henneguya spp. em peixes criados no Sudoeste de Goiás.

Three outbreaks of infection by Piscinoodinium pillulare and Henneguya spp. in Piarac tus mesopotamicus intensively raised in Southwestern Goiás, Brazil, are described. Two outbreaks occurred in the winter and another one in summer. Morbidity (58.57%-90%) and mortality (100%) rates were determined. Stress episodes occurred previously in all cases. Clinical signs included behavior changes and erratic movements with loss of equilibrium. Main gross findings were swelling of the gills with white mucus exudate. Microscopically, gills had mild to moderate multifocal necrotizing and lympho-plasmocytic inflammation with hypertrophy and fusion of secondary lamellae. In addition, numerous intralesional trophonts of P. pillulare and cysts of Henneguya spp. were noted. According to our knowledge, this seems to be the first description of Piscinoodinium pillulare e Henneguya spp. infection in fishes reared in Southwestern Goiás, Brazil.

Intoxicação espontânea por Senna occidentalis em javalis (Sus scrofa ferus) no estado de Goiás / Spontaneous poisoning by Senna occidentalis in wild boars (Sus scrofa ferus) in Goiás, Brazil

Descreve-se um surto de intoxicação por Senna occidentalis em javalis no Estado de Goiás. De um rebanho de 80 javalis, 15 adoeceram e um morreu. Os sinais clínicos observados foram apatia, prostração, ataxia, tremores musculares, incoordenação, relutância em mover-se, decúbito esternal ou lateral, paresia e paralisia espástica, principalmente dos membros pélvicos. As principais alterações macroscópicas consistiam de palidez moderada a acentuada da musculatura esquelética dos membros pélvicos e torácicos e no lombo, especialmente nos músculos da coxa. Microscopicamente, observou-se degeneração e necrose flocular, multifocal, leve a moderada, monofásica, com fragmentação de fibras na musculatura esquelética. Nos cortes transversais, havia fibras musculares tumefeitas e hipereosinofílicas. Adicionalmente, havia degeneração microvacuolar hepatocelular difusa, leve a moderada. A atividade sérica da CK estava acentuadamente elevada em dois javalis avaliados e da TGO aumentou em um javali afetado.

An outbreak of poisoning by Senna occidentalis in wild boars in Goiás, Brazil, is described. Out of 80 wild boars, 15 were affected and one died. Clinical signs included apathy, muscle tremors, incoordination, reluctance to move, sternal or lateral recumbence, and paresis and spastic paralysis, especially in the hind limbs. Gross lesions were characterized by moderate to severe paleness of the skeletal muscles of the hind and thoracic limbs and loin, and were marked in the thigh muscles. Histologically, a monophasic, mild to moderate, multifocal floccular muscle degeneration and necrosis with skeletal fiber sarcoplasmic fragmentation was observed. In transverse sections there were multiple swollen and hypereosinophilic muscle fibers. In addition, mild to moderate, diffuse microvacuolar degeneration was seen in the liver. Creatine phosphokinase serum activity was severely elevated in two evaluated wild boars. In one of them, there was elevation of glutamic oxaloacetic transaminase serum activity.

Polioencefalomalacia experimental induzida por amprólio em bovinos / Experimentally amprolium-induced polio-encephalomalacia in cattle

Para estabelecer um modelo experimental para o estudo da etiologia, patologia e patogênese da polioencefalomalacia (PEM) em bovinos, a condição foi induzida em quatro novilhos pela administração oral de amprólio nas doses diárias de 500 e 350mg/kg de peso vivo, respectivamente por 22 e 26-28 dias. Todos os bovinos morreram espontaneamente ou foram eutanasiados in extremis após um curso clínico de 4-7 dias. Três bovinos que receberam 1.000mg/kg de amprólio e dois que receberam 500mg/kg morreram espontaneamente com quadro clínico agudo a subagudo sem desenvolverem sinais e lesões de PEM. Nos novilhos que PEM foi reproduzida, os sinais neurológicos incluíram marcada apatia, incoordenação, posição de cavalete, quedas ocasionais, hiperexcitabilidade, tremores musculares, cegueira, bruxismo, estrabismo, nistagmo, midríase, opistótono, decúbito lateral e movimentos de pedalagem. Os principais achados de necropsia eram restritos ao encéfalo e consistiam de tumefação, achatamento, amolecimento e amarelamento das circunvoluções cerebrais. Histologicamente, havia necrose neuronal segmentar e laminar (neurônios vermelhos) associada a edema, tumefação endotelial, separação das lâminas de neurônios do córtex telencefálico ou entre as substâncias cinzenta e branca e infiltração moderada a acentuada de macrófagos espumosos. Essas alterações eram mais acentuadas nos lobos telencefálicos frontal, parietal e occipital. Adicionalmente, lesões similares e moderadas foram detectadas no mesencéfalo e hipocampo. A necrose neuronal e o edema afetaram uniformemente as camadas de neurônios da substância cinzenta dos lobos telencefálicos frontal, parietal e occipital. Esse modelo experimental de PEM com administração oral de amprólio parece ser útil para o estudo da doença em bovinos, conforme observado anteriormente em ovinos.

In order to establish an experimental model for the study of the etiology, pathology, and pathogenesis of polioencephalomalacia (PEM) in cattle, the condition was induced into four steers by oral administration of amprolium at daily doses of 500 and 350mg per kg of body weight respectively for 22 and 26-28 days. All steers died spontaneously or were euthanized in extremis after being sick for 4-7 days. Three steers that received the drug at 1,000mg/kg and two that received 500mg/kg died spontaneously with acute or subacute clinical signs and without lesions and signs of PEM. In those steers in which PEM was reproduced, the neurological signs included marked apathy, incoordination, sawhorse stance, occasional falls, hyperexcitability, muscle tremors, blindness, grinding of teeth, strabismus, nystagmus, mydriasis, opisthotonus, and lateral recumbency with paddling movements. Main gross lesions were restricted to the brain and included swelling, flattening, softening and yellow discoloration of the cerebral circumvolutions. Histologically, there was segmental laminar neuronal necrosis (red neurons) associated with edema, swelling of endothelial cells, cleavage of laminar neuronal layers or between gray and white matter and moderate to severe infiltration by foamy macrophages (gitter cells). These changes were more marked in the frontal, parietal and occipital telencephalic lobes. Additionally, similar and moderate lesions were detected in the midbrain and hippocampus. Neuronal necrosis and edema affected uniformly the neurons layers of the grey matter of the frontal, parietal and occipital lobes. This experimental model of PEM with oral administration of amprolium may be useful for the study in cattle, as previously observed in sheep.

Polioencefalomalacia experimental induzida por amprólio em ovinos / Experimentally amprolium-induced polioencephalomalacia in sheep

Para estabelecer um modelo experimental para o estudo da etiologia, patologia e patogênese da polioencefalomalacia em ruminantes, a condição foi induzida em cinco ovinos pela administração oral de amprólio nas doses diárias de 500 e 1.000mg/kg de peso animal, respectivamente por 28-59 dias e 13-39 dias. Todos os ovinos morreram ou foram eutanasiados in extremis após um curso clínico de 3-7 dias. Os sinais clínicos incluíam depressão, incoordenação, midríase, bruxismo, cegueira e decúbito com opistótono e movimentos de pedalagem. Salivação excessiva e posição de cavalete foi observada em um ovino e mioclonias em um outro. Os principais achados de necropsia restringiam-se ao sistema nervoso central e incluíam tumefação do encéfalo com achatamento dos giros telencefálicos e hemorragias nos lobos parietal e occipital do telencéfalo; as hemorragias ocorriam também nas áreas submeníngeas da medula espinhal e do mesencéfalo. Histologicamente, havia necrose segmentar laminar de neurônios (neurônios vermelhos) associada a edema, tumefação de células endoteliais, hemorragias e infiltração por macrófagos espumosos (células gitter). Essas alterações eram mais marcadas nos lobos frontal, parietal e occipital do telencéfalo e havia uma demarcação abrupta entre as lesões e o neurópilo normal adjacente. Adicionalmente, lesões semelhantes, mas menos acentuadas, eram observadas no mesencéfalo, tálamo e hipocampo. Levando em consideração a reproducibilidade regular dos aspectos da polioencefalomalacia em ovinos pela administração de amprólio, esse modelo pode ser útil para o estudo da doença.

In order to establish an experimental model for the study of the etiology, pathology, and pathogenesis of polioencephalomalacia in ruminants, the condition was induced in five sheep by oral administration of amprolium at daily doses of 500 and 1,000mg per kg of body weight respectively for 28-59 days and for 13-39 days. All sheep died or were euthanized in extremis after illness of 3-7 days. Clinical signs included depression, incoordination, midriasis, grinding of the teeth, blindness, and laying down with opisthotonus and paddling movements. Drooling and a sawhorse stance were observed in one sheep and myoclonus in another one. Main gross lesions were restricted to the central nervous system and included swelling of the brain with flattening of telencephalic gyri, and hemorrhages in the parietal and occipital lobes of the telencephalon, in the submeningeal areas of the spinal cord and in the mesencephalon. Histologically, there was segmental laminar neuronal necrosis (red neurons) associated with edema, swelling of endothelial cells, hemorrhages and infiltration by foamy macrophages (gitter cells). These changes were more marked in the frontal, parietal and occipital telecephalic lobes and there was sharp demarcation between the lesions and the adjacent normal neuropile. Additionally, similar, but less marked lesions were seen in the mesencephalon, thalamus and hippocampus. Considering the consistent reproducible aspects of polioencephalomalacia in sheep using amprolium, this may be an useful model for the study of the disease.