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- A Nationwide Seroepidemiologic Study on Q Fever Antibodies in Sheep of PortugalPublication . Cruz, Rita; Esteves, Fernando; Nóbrega, Carmen; Santos, Carla; Ferreira, Ana S.; Mega, Cristina; Coelho, Ana C.; Vala, Helena; Mesquita, JoãoIntroduction: Q fever is an almost global zoonotic disease caused by Coxiella burnetii. Human infections can produce acute and chronic disease that can lead to abortions and stillbirths in pregnant women, usually infected by the inhalation of C. burnetii-contaminated aerosols or through consumption of contaminated products. Sheep are one of the primary animal reservoirs with disease being associated with vast shedding of bacteria in placentas, feces, milk, and birth fluids. Although almost neglected in the past, recent outbreaks of sheep origin have alerted the public and the scientific community. Materials and Methods: An epidemiologic survey to estimate the seroprevalence of Q fever antibodies was performed in a representative number of sheep of all regions of continental Portugal (n = 1068), using a commercial ELISA (ID Screen Q Fever Indirect Multi-species Kit; IDvet, Montpellier, France). Results and Discussion: An anti-C. burnetii seroprevalence of 11.4% (95% confidence interval 9.6–13.5) was found, with a clear distinction between the Center region with highest seroprevalence, and the rest of the territory. Sheep traditional farming is widely present in Portugal and is part of the cultural and gastronomical background of the country. This close proximity to small ruminants may contribute to the zoonotic transfer to humans.
- Hepatic histopathological lesions in acute controled haemorrhage followed by volume replacement with a crystalloid or colloid solutionPublication . Vala, Helena; Pina, R.; Cruz, R.; Venâncio, C.; Esteves, Fernando; Silva, A.; Mesquita, João; Ortiz, A. L.; Ferreira, D.Introduction: severe hemorrhage remains the major cause of morbidity and mortality in trauma victims or surgical intervened animals, despite of all the advances in the therapeutic approach. The resulting injuries, or even death, are due to the deficit in intravascular volume and subsequent development of hypovolaemic fluid state, leading to poor tissue perfusion and consequent decreased oxygen delivery to the tissues, often with compromise of organ function. Objective: the aim of this work was to evaluate the effect of different intravenous solutions used for volume replacement following acute controlled haemorrhage, one crystalloid (RL) and one colloid (HES130/0.4) in the integrity of the hepatic tissue in a pig animal model. Material and methods: hepatic samples were collected from animals submitted to passive arterial blood bleeding and reperfusion with a crystalloid (RL) (G1) and with a synthetic colloid (HES 130/0.4) (G2). Samples were also collected from animals that were not subjected to acute bleeding nor volume replacement (G3, control group). All procedures were carried out under personal and project licenses approved by the Ethical Committee of the national regulatory office. Samples were collected and fixed in 10% neutral-buffered formalin, for a maximum of 24hours, embedded in paraffin wax and 3µm sections were stained for routine histopathology with haematoxylin and eosin. Results: the histopathological assessment revealed no statistically significant differences between the three groups. However, some lesions were more often expressed in some groups. More severe hepatocellular hydropic degeneration and hepatocellular steatosis was seen in G1, which is the only group in which haemorrhage was observed and within which oedema was not present. Hyperaemia was only observed in G2 and G3. Necrosis was not present in any of the groups. Conclusion: hepatic histopathological lesions following controlled bleeding and intravenous volume replacement with RL or HES130/0.4 were subtle. However, more pronounced hydropic degeneration and hepatocellular steatosis was seen in G1 (RL), which suggests that HES130/04 may be associated with better hepatic perfusion when used for intravenous volume replacement when compared to RL.
- Crimean-Congo hemorrhagic fever virus circulating among sheep of Portugal: a nationwide serosurvey assessmentPublication . Mesquita, João; Cruz, Rita; Esteves, Fernando; Santos, Carla; Pousa, Humberto; Coelho, Catarina; Mega, Cristina; Nóbrega, Carmen; Vala, Helena; Peyrefitte, Christophe Nicolas; Nascimento, Maria São José; Barradas, Patrícia FerreiraCrimean-Congo hemorrhagic fever virus (CCHFV) is a widespread zoonotic pathogen that can cause mild to severe hemorrhagic disease in humans. CCHFV may be transmitted through direct contact with tissue or blood of viremic animals; however, the primary transmission route is through infected tick bites. CCHFV RNA has been detected in ticks feeding on domestic and wild animals in western Spain, suggesting an established circulation of CCHFV in Western Europe. Ruminants have been recognized as important CCHFV reservoirs and have been linked to human cases in endemic regions. Given the emergence of CCHF in neighboring Spain, and a report of two CCHFV seropositive humans in southern Portugal in 1985, we investigated the potential circulation of this virus in the country by performing a nationwide anti-CCHFV IgG serosurvey in sentinel sheep of Portugal. Sera (n = 459) randomly selected from widely distributed farms (n = 20) of Portugal were tested using a commercial double-antigen enzyme-linked immunosorbent assay, yielding an overall seroprevalence of 0.4% (95% confidence interval [CI] 0.04-1.56%). Positive sheep were from the southern region of Portugal (Alentejo region), which raise the seroprevalence of this region to 0.74% (95% CI 0.09-2.66%). This is the first study reporting the presence of CCHFV antibodies in sheep of Portugal, thus suggesting a geographical expansion of CCHFV to this country. It seems likely that CCHFV may exist focally in southern Portugal.
- Inspecção do sistema nervoso central em pequenos ruminantesPublication . Vala, Helena; Mesquita, João; Esteves, Fernando; Pinho, FátimaO Scrapie é uma doença neurodegenerativa fatal que afecta ovinos e caprinos e é a forma mais comum das Encefalopatias Espongiformes Transmissíveis (EET's), que incluem a doença de Creutzfeldt – Jakob (CJD), em humanos, e a Encefalopatia Espongiforme Bovina (BSE). O Plano de Vigilância Activa do Scrapie, em aplicação na União Europeia desde 2002, permitiu determinar uma prevalência de Scrapie superior ao esperado. De forma a efectuar uma avaliação preliminar da incidência desta patologia em Portugal, foram recolhidos os troncos cerebrais e cerebelos de 4822 pequenos ruminantes abatidos num matadouro no centro de Portugal e, posteriormente, enviados para pesquisa laboratorial de EETs. Após análise laboratorial, detectaram-se 8 casos de Scrapie atípico em ovinos. A percentagem de casos positivos (0,17%) detectados em pequenos ruminantes com mais de 18 meses, durante o período de realização deste estudo, foi baixa e, segundo os resultados laboratoriais, enquadram-se no perfil de Scrapie atípico.
- A case of vascular tumour mimicking lymphangiomaPublication . Vala, Helena; Mega, Cristina; Santos, C.; Cruz, Rita; Santos, M.; Esteves, Fernando; Nóbrega, Carmen; Mesquita, JoãoIntroduction Sarcomas, particularly vascular tumours, can mimic other neoplastic conditions or even inflammatory diseases. Objectives The authors describe an unusual case of vascular tumour, histologically overlapping with benign lymphangioma and liposarcoma. Medical History A 8 year old Shar Pei male dog was presented with a subcutaneous nodule in the chest, measuring 3x2 cm. Material and Methods The sample was fixed in 10 % buffered formalin solution and processed for routine histopathological diagnosis with Haematoxylin and Eosin. A standard Streptavidin-Biotin Complex technique for immunohistochemical evaluation was also performed, using a Polyclonal Rabbit Anti-Human Von Willebrand Factor. Results Macroscopic examination revealed a well circumscribed nodule with homogeneous and brownish appearance and moderate consistency on a cut surface. Microscopic examination revealed clefts and variably-sized vascular spaces, lined by a single layer of spindle cells, with round-to-oval nuclei, small or indistinct nucleoli, small amount of eosinophilic cytoplasm, with endothelial appearance – areas like hemangiosarcoma, interspersed with areas, where these cells appeared to grow directly on thick bundles of dermal collagen, occasionally with erythrocytes – areas like lymphangioma. The stroma was also interspersed with vacuolar cells – areas like liposarcoma. No mitotic figures were seen, however, the presence of very rich cellular areas, with more prominent cells revealing pleomorphism and anisocariosis, were more favour to malignancy Conclusion The authors suggested the diagnosis of vascular neoplasia with unusual features, based on the histopathological diagnosis and immunohistochemistry results, and submit to the discussion, especially for assessment of histological grade and prognosis.
- Epidemiology and Emergence of Schmallenberg Virus Part 1: Origin, Transmission and Differential DiagnosisPublication . Esteves, Fernando; Mesquita, João; Nóbrega, Carmen; Santos, Carla; Monteiro, António; Cruz, Rita; Vala, Helena; Coelho, Ana CláudiaSchmallenberg virus (SBV) is a novel Orthobunyavirus causing mild clinical signs in cows and malformations in aborted and neonatal ruminants in Europe. SBV belongs to the family Bunyaviridae and is transmitted by biting midges. This new virus was identified for the first time in the blood samples of cows in the city of Schmallenberg in NorthRhine-Westphalia in November 2011. Since then the virus spread to several European countries. Here we describe the origin and emergence, as well as the transmission and the differential diagnosis of this virus, now known to be a serious threat to Veterinary Public Health.
- Bioética e experimentação animalPublication . Nóbrega, Carmen; Oliveira, P. A.; Mesquita, João; Santos, C.; Arantes, R.; Themudo, D.; Esteves, Fernando; Cruz, Rita; Mega, Cristina; Vala, Helena- Introdução: O contributo da experimentação animal para a ciência é inegável. Dificilmente os meios médicos e profiláticos teriam atingido o presente estado de evolução sem a sua contribuição. O objectivo deste trabalho é a apresentação do conceito de experimentação animal, como uma prática válida e eticamente correta, desde que devidamente planeada e executada. - Estado da Arte: O recurso a animais vivos em investigação, tem desempenhado um papel importante no avanço científico, particularmente na área da saúde. Este facto dá origem a um dilema ético: se por um lado a maior parte dos animais utilizados são seres sencientes, por outro lado, a investigação pode ser de vital importância para a prevenção, cura ou melhoria significativa de doenças em humanos. Não sendo um tema consensual, é geralmente alvo de intenso debate. Opositores a qualquer tipo de investigação com recurso a animais, afirmam que a experimentação animal é cruel e desnecessária porque os animais não existem para serem usados segundo o nosso critério. No entanto, a generalidade da sociedade aceita que os animais possam ser utilizados em experimentação mas não a qualquer preço. Nos últimos anos, têm ocorrido progressos na criação de alternativas à experimentação animal, nomeadamente as culturas celulares, de tecidos e órgãos, fazendo com que, em certas situações, os animais possam já ser substituídos por alternativas eticamente mais aceitáveis, não podem, contudo, ser inteiramente eliminados da investigação. - Novas perspetivas/diretrizes Uma investigação ética e cientificamente válida com aplicabilidade na saúde humana, exige a seleção de modelos adequados para a acção estudada e o cumprimento do princípio dos 3Rs: Replace; Reduce; Refine. A estes, deve juntar-se ainda o R da responsabilidade, que deve prevalecer desde o delineamento da experimentação e acompanhar o decurso dos trabalhos experimentais. - Implicações teóricas e práticas Pesando os prós e contras, os benefícios da experimentação animal pesam mais no que se refere ao impacto na melhoria da saúde humana. Se esta prática fosse abandonada, certamente existiriam severas consequências a nível da investigação e da saúde pública.
- Increased risk of hepatitis E virus infection in workers occupationally exposed to sheepPublication . Mesquita, João; Santos-Ferreira, Nanci; Ferreira, Ana S.; Albuquerque, Carlos; Nóbrega, Carmen; Esteves, Fernando; Cruz, Rita; Vala, Helena; Nascimento, Maria S. J.Hepatitis E virus (HEV) is an enteric RNA virus from the family Hepeviridae with five genotypes (genotypes 1–4 and 7) known to infect humans. HEV infection is known to have a zoonotic swine origin in industrialized countries. The role of pigs and wild boars as major reservoirs for human infection is today well-established; however, the list of new animal reservoirs is ever-expanding as new HEV strains are continuously being found in a broad host range. The recent detection of HEV in sheep stools brings concerns on the possibility of HEV transmission from these animals to humans, particularly in those occupationally exposed. The present work investigated the potential occupational risk of HEV infection in shepherds and sheep milk cheesemakers—workers occupationally exposed to ovine (WOEOs; N = 96)—from a region of the Centre of Portugal (‘Serra da Estrela’) based on the differences of anti-HEV IgG seroprevalence rates between these professionals and the general population (N = 192). The presence of HEV-specific antibodies in sheep (N = 90) from the same region was also evaluated. The HEV seroprevalence in WOEOs (29.3%) was found to be significantly higher (p = .0198) when compared with population controls (16.1%) which suggests an increased risk for HEV infection in these workers. HEV-specific antibodies were also found in 16.6% of the studied sheep showing that HEV circulates in these animals. Further studies are needed to confirm the zoonotic potential of sheep HEV.
- Mammary neoplasms in dogs and cats. Contribution for promotion of well-beingPublication . Soares, Marta; Almeida, Rui Gomes; Esteves, Fernando; Cruz, Rita; Santos, Carla; Mega, Cristina; Nóbrega, Carmen; Mesquita, João; Vala, HelenaIntrodução: Nos últimos anos tem-se verificado um crescimento significativo da incidência de neoplasias humanas a nível mundial, sendo esta também uma das principais causas de morbilidade e mortalidade nos animais de companhia, destacando-se as neoplasias mamárias como processo neoplásico que mais acomete os nossos animais. Objetivos: Analisar o paradigma atual das neoplasias em animais de companhia; Intervir na sensibilização de tutores e profissionais da equipa de saúde animal para uma gestão mais assertiva, nomeadamente no que respeita à sua deteção e exérese precoce. Métodos: estudo retrospetivo contemplando uma amostragem de canídeos (n=20) e felídeos (n=12) com massas mamárias submetidas a excisão cirúrgica e diagnóstico histopatológico. Resultados: Na amostra analisada foi observada uma maior frequência de neoplasias mamárias em cadelas de raça Indeterminada e em gatas de raça Europeu Comum, sendo a idade média, para ambas as espécies, próxima dos 10 anos de idade. Foi ainda registada uma maior frequência de neoplasias mamárias malignas, 75% em cadelas e 66,7% nas gatas, em relação às benignas. Conclusões: Foi possível verificar que na região do país contemplada neste estudo foram diagnosticadas mais neoplasias mamárias malignas do que benignas, provavelmente motivado pela excisão cirúrgica tardia de massas mamárias grandes, por vezes ulceradas.
- Occurrence and Distribution of Fasciolosis in a Cohort of Ovine Livestock Restricted to a Mountain Plateau in Central PortugalPublication . Coelho, Catarina; Cruz, Rita; Esteves, Fernando; Vala, Helena; Pereira, Maria A.; Amorim, Irina; Nóbrega, Carmen; Mesquita, JoãoFasciola hepatica is a parasite that is widespread in Europe, having been reported in ru- minants of several European countries and causing an important economic impact. This study ascertained the occurrence and distribution of fasciolosis in Portuguese ovine livestock by assessing F. hepatica IgG antibodies in a cohort of confined sheep from a high-altitude region of central Portugal in a 2-year period. Positive animals were found in most locations and in both years, with 18 of the 92 an- imals (19.6% [95% confidence interval CI: 12.03–19.15]) and 17 of the same 92 animals (18.5% [95% CI: 11.15–27.93]) showing to be seropositive in the first year and second year, respectively (p = 0.85). Pasture contamination by F. hepatica eggs could be reduced by thorough anthelmintic treatments.
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