Browsing by Author "Ramos, Fernanda"
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- Simple method for establishing primary Leporidae skin fibroblast culturesPublication . Abade dos Santos, F.A.; Carvalho, C. L.; Almeida, Isabel; FAGULHA, TERESA; Ramos, Fernanda; Barros, Sílvia C.; Henriques, Margarida; Luís, Tiago; Duarte, Margarida D.Commercial hare and rabbit immortalized cell lines are extremely limited regarding the many species within the lagomorpha order. To overcome this limitation, researchers and technicians must establish primary cell cultures derived from biopsies or embryos. Among all cell types, fibroblasts are plastic and resilient cells, highly convenient for clinical and fundamental research but also for diagnosis, particularly for viral isolation. Here, we describe a fast and cheap method to produce primary fibroblast cell cultures from leporid species, using dispase II, a protease that allows dermal–epidermal separation, followed by a simple enzymatic digestion with trypsin. This method allows for the establishment of an in vitro cell culture system with an excellent viability yield and purity level higher than 85% and enables the maintenance and even immortalization of leporid fibroblastic cells derived from tissues already differentiated.
- West Nile virus transmission potential in PortugalPublication . Lourenço, José; Barros, Sílvia C.; Zé-Zé, Líbia; Damineli, Daniel S. C.; Giovanetti, Marta; Osório, Hugo; Amaro, Fátima; Henriques, Ana M.; Ramos, Fernanda; Luís, Tiago; Duarte, Margarida D.; Fagulha, Teresa; Alves, Maria João; Obolski, UriIt is unclear whether West Nile virus (WNV) circulates endemically in Portugal. Despite the country's adequate climate for transmission, Portugal has only reported four human WNV infections so far. We performed a review of WNV-related data (1966-2020), explored mosquito (2016-2019) and land type distributions (1992-2019), and used climate data (1981-2019) to estimate WNV transmission suitability in Portugal. Serological and molecular evidence of WNV circulation from animals and vectors was largely restricted to the south. Land type and climate-driven transmission suitability distributions, but not the distribution of WNV-capable vectors, were compatible with the North-South divide present in serological and molecular evidence of WNV circulation. Our study offers a comprehensive, data-informed perspective and review on the past epidemiology, surveillance and climate-driven transmission suitability of WNV in Portugal, highlighting the south as a subregion of importance. Given the recent WNV outbreaks across Europe, our results support a timely change towards local, active surveillance.