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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.
- Outbreaks of abortions byCoxiella burnetiiin small ruminant flocks and a longitudinal serological approach on archived bulk tank milk suggest Q fever emergence in Central PortugalPublication . Cruz, Rita; Esteves, Fernando; Vasconcelos-Nóbrega, Carmen; Santos, Carla; Ferreira, Ana S.; Mega, Cristina; Coelho, Ana C.; H, Vala; Mesquita, João R.Q fever is a worldwide zoonotic infectious disease caused by Coxiella burnetii and sheep and goats are known to be the main reservoir for human infection. This study describes the epidemiological and laboratory findings of C. burnetii outbreaks affecting sheep and goat flocks and also provides the results of a prospective serosurvey in bulk tank milk samples to assess C. burnetii circulation in a population of sheep living in close contact to the human population in Central Portugal. In the epizooties, C. burnetii was identified in tissues of the resulting abortions by qPCR. As for the serological survey, 10.2% (95%CI: 4.5‐19.2) of the 78 bulk tank milk samples collected in 2015 presented IgG antibodies against C. burnetii. The same farms were visited and sampled in 2016 and 25.6% (95%CI: 16.4‐36.8) were positive. This steep increase in the number of anti‐C. burnetii farms between the 2015 and 2016 collections showed to be statistically significant (p = 0.020) and is strongly suggestive of Q fever emergence in Central Portugal. Measures on animal health and on disease spread control to the human population should be considered.
- A 2-Year Longitudinal Seroepidemiological Evaluation of Toxoplasma gondii Antibodies in a Cohort of Autochthonous Sheep from Central PortugalPublication . Almeida, Daniela; Quirino, João; Barradas, Patrícia Ferreira; da Silva, Priscilla Gomes; Pereira, Maria; Cruz, Rita; Santos, Carla; Mega, Cristina; Esteves, Fernando; Nóbrega, Carmen; Vala, Helena; Gärtner, Fátima; Amorim, Irina; Mesquita, João R.Background: Toxoplasma gondii is an important zoonosis and one of the major causes of abortion in sheep worldwide. (2) Methods: We performed a 2-year longitudinal serological antiT. gondii IgG screening on a cohort of a spatially confined population of a Portuguese autochthonous sheep breed in central Portugal. (3) Results: From the screening of the 2015 and 2016 sera, an increase of seroprevalence was observed (57.7% (95% CI: 49.9–65.3%) versus 69.1% (95% CI: 61.5–75.9), from 2015 and 2016, respectively) (p = 0.031). (4) Conclusions The present study is the first to provide prospective data on the anti-T. gondii serological status of a sheep cohort in Portugal, showing an increase in the occurrence of T. gondii. There is a need to provide a clearer understanding of T. gondii epidemiology in Portugal, ideally by implementing monitoring programs on sentinel herds, not only due to the high impact of T. gondii on animal health but also for it being a zoonosis.
- A Questionnaire-Based Survey on the Long-Term Management of Canine Leishmaniosis by Veterinary PractitionersPublication . Pereira, Maria; Santos, Rute; Nóbrega, Carmen; Mega, Cristina; Cruz, Rita; Esteves, Fernando; Santos, Carla; Mesquita, João R.; Vala, Helena; Santos-Gomes, GabrielaCanine Leishmaniosis (CanL) is a chronic and potentially fatal disease. In economically disadvantaged regions, costs associated with long-term patient monitoring may determine that some owners decline veterinary follow-up of their dogs. This online, questionnaire-based survey aimed to assess how Portuguese veterinary practitioners perform long-term patient monitoring and recognize relapses. More than 50% of respondents reported that 50–100% of dog owners declared financial restraints. Hence, in these circumstances, most veterinary practitioners only performed clinical examination and serology. However, when owners did not declare financial restriction, other tests were additionally performed, such as renal and hepatic profiles, hemogram, serum protein electrophoresis and urine protein creatinine ratio. The mean number of exams performed when owners presented financial restraints was significantly lower than the number of exams performed without economic limitations. Most veterinary practitioners prescribed allopurinol ad aeternum oruntil disease remission and domperidone. CanL relapses were recognized by more than half of respondents “Always”, through the reappearance or worsening of clinical signs, whereas about a quarter detected an increase in anti-Leishmania antibody levels and identified abnormalities in the serum protein electrophoresis profile. The relapse rate was higher in the Lisbon Metropolitan Area and north, the most economically favored regions of Portugal. This study confirms that owner financial restraints negatively influence veterinary follow-up and relapse recognition, ultimately compromising clinical decision making and favoring the maintenance of Leishmania infantum infection endemicity.
- 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.
- 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.
- Pulmonary and Gastrointestinal Parasitic Infections in Small Ruminant Autochthonous Breeds from Centre Region of Portugal—A Cross Sectional StudyPublication . Pereira, Maria; Vila-Viçosa, Maria; Coelho, Catarina; Santos, Carla; Esteves, Fernando; Cruz, Rita; Gomes, Liliana; Henriques, Diogo; Vala, Helena; Nóbrega, Carmen; Mega, Cristina; Melo, Carolina de; Malva, Madalena; Braguez, Joana; Mateus, Teresa LetraThe production of small ruminant autochthonous breeds in the Centre region of Portugal is practiced in a semi-extensive husbandry system, exposing animals to parasitic infections. The main objective of this study was to estimate the prevalence of lungworm infection and identify risk factors. Fecal samples of 203 goats and 208 sheep from 30 herds were collected per rectum and subjected to the modified Baermann test. The overall prevalence of infection was 57.7%, significantly higher in goats (95.6%) than in sheep (20.7%) (p < 0.001). According to the binary logistic regression model, sheep dewormed with albendazole, mebendazole plus closantel, or ivermectin plus clorsulon presented a risk of Protostrongylidae infection 29.702, 7.426, or 8.720 times higher, respectively, than those dewormed with eprinomectin. Additionally, the presence of gastrointestinal parasites was investigated in 307 fecal samples using Mini-FLOTAC®. The overall prevalence of infection was 86.3%, also significantly higher in goats (93.2%) than in sheep (79.9%) (p < 0.001). Strongyle-type eggs were the most frequently identified, both in sheep (69.8%) and goats (87.8%), followed by Eimeria oocysts (40.3% in sheep and 68.9% in goats). Considering the high prevalence and the burden of lungworm parasitic infection, it is urgent to determine its economic impact and the repercussions in animal health in the Centre region of Portugal to establish appropriate therapeutic guidelines.
- Occupationally Acquired Q Fever in Shepherds and Sheep Milk CheesemakersPublication . Cruz, Rita; Santos, Carla; Esteves, Fernando; Ferreira, Ana Sofia; Nóbrega, Carmen; Mega, Cristina; Albuquerque, Carlos; Teixeira-de-Lemos, Edite; Coelho, Ana Cláudia; Vala, Helena; Mesquita, JoãoQ fever is a zoonosis caused byCoxiella burnetii, and transmission to humans is often associatedwith contact with ovine and caprine livestock. Those exposed to sheep are particularly at high risk of infection.Recent studies show that Q fever is increasing in sheep farms in Portugal raising alerts on spillover to humans.We detected anti-C. burnetiiIgG in shepherds and sheep milk cheesemakers (27 [28.1%] in a total of 96; 95%confidence interval [CI] 19.4–38.2%) and in controls (21 [8.1%] in a total of 260; 95% CI 5.1–12.1%), pointingto an increased risk ofC. burnetiiinfection (P= 0.0001), with an odds ratio foranti-C. burnetiiof 4.45 (95% CI2.4–8.4%;P= 0.0001), in individuals with occupational contact with sheep in Portugal.
- 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.
- 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.
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