Browsing by Author "Parreira, Ricardo"
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- A diverse assemblage of RNA and DNA viruses found in mosquitoes collected in southern PortugalPublication . Silva, Manuel; Morais, Paulo; Maia, Carla; Sousa, Carolina Bruno de; Gouveia de Almeida, Antonio Paulo; Parreira, RicardoThis work describes the detection and partial characterization of mosquito-borne virus genomic sequences, based on the analysis of mosquitoes collected from the Spring to Fall of 2018 in the Algarve (southern Portugal). The viral survey that was carried out using multiple primer sets disclosed the presence of both RNA and DNA viral sequences in these mosquitoes, which were subsequently analysed using maximum likelihood and Bayesian phylogenetic reconstruction methods. The obtained results brought to light three lineages of insect-specific flaviviruses, a monophyletic cluster of bunyaviruses from an unassigned group within the Phenuiviridae family, as well as brevidensoviruses (Parvoviridae, Densovirinae:). The latter two groups of viruses were here described for the first time in mosquitoes from Portugal. Results relating to the tentative isolation of the putative viruses identified in C6/36 cells are also shown, and the serendipitous, although not unexpected, isolation a Negev-like Nelorpivirus from Culex laticinctcus mosquitoes is reported.
- Molecular detection of Leishmania DNA and identification of blood meals in wild caught phlebotomine sand flies (Diptera: Psychodidae) from southern PortugalPublication . Maia, Carla; Parreira, Ricardo; Cristovao, Jose Manuel; Freitas, Ferdinando Bernardino; Afonso, Maria Odete; Campino, LeneaBackground: Zoonotic visceral leishmaniasis caused by Leishmania infantum which is transmitted by phlebotomine sand flies (Diptera, Psychodidae) is endemic in the Mediterranean basin. The main objectives of this study were to (i) detect Leishmania DNA and (ii) identify blood meal sources in wild caught female sand flies in the zoonotic leishmaniasis region of Algarve, Portugal/Southwestern Europe. Methods: Phlebotomine sand flies were collected using CDC miniature light traps and sticky papers. Sand flies were identified morphologically and tested for Leishmania sp. by PCR using ITS-1 as the target sequence. The source of blood meal of the engorged females was determined using the cyt-b sequence. Results: Out of the 4,971 (2,584 males and 2,387 females) collected sand flies, Leishmania DNA was detected by PCR in three females (0.13%), specifically in two specimens identified on the basis of morphological features as Sergentomyia minuta and one as Phlebotomus perniciosus. Haematic preferences, as defined by the analysis of cyt-b DNA amplified from the blood-meals detected in the engorged female specimens, showed that P. perniciosus fed on a wide range of domestic animals while human and lizard DNA was detected in engorged S. minuta. Conclusions: The anthropophilic behavior of S. minuta together with the detection of Leishmania DNA highlights the need to determine the role played by this species in the transmission of Leishmania parasites to humans. In addition, on-going surveillance on Leishmania vectors is crucial as the increased migration and travelling flow elevate the risk of introduction and spread of infections by Leishmania species which are non-endemic.
- Molecular detection of tick-borne bacteria and protozoa in cervids and wild boars from PortugalPublication . Pereira, André; Parreira, Ricardo; Nunes, Monica; Casadinho, Afonso; Vieira, Maria Luisa; Campino, Lenea; Maia, CarlaBackground: Wildlife can act as reservoir of different tick-borne pathogens, such as bacteria, parasites and viruses. The aim of the present study was to assess the presence of tick-borne bacteria and protozoa with veterinary and zoonotic importance in cervids and wild boars from the Centre and South of Portugal.Methods: One hundred and forty one blood samples from free-ranging ungulates including 73 red deer (Cervus elaphus), 65 wild boars (Sus scrofa) and three fallow deer (Dama dama) were tested for the presence of Anaplasma marginale/A. ovis, A. phagocytophilum, Anaplasma/Ehrlichia spp., Babesia/Theileria spp., Borrelia burgdorferi (sensu lato) (s.l.), and Rickettsia spp. DNA by PCR.Results: Anaplasma spp. DNA was detected in 33 (43.4 %) cervids (31 red deer and two fallow deer) and in two (3.1 %) wild boars while Theileria spp. were found in 34 (44.7 %) cervids (32 red deer and two fallow deer) and in three (4.6 %) wild boar blood samples. Sequence analysis of msp4 sequences identified A. marginale, A. ovis, while the analysis of rDNA sequence data disclosed the presence of A. platys and A. phagocytophilum and T. capreoli and Theileria sp. OT3. Anaplasma spp./Theileria spp. mixed infections were found in 17 cervids (22.4 %) and in two wild boars (3.1 %). All samples were negative for Babesia sp., B. burgdorferi (s.l.), Ehrlichia sp. or Rickettsia sp.Conclusions: This is the first detection of Anaplasma marginale, A. ovis, A. phagocytophilum, A. platys, Theileria capreoli and Theileria sp. OT3 in cervids and wild boars from Portugal. Further studies concerning the potential pathogenicity of the different species of Anaplasma and Theileria infecting wild ungulates, the identification of their vector range, and their putative infectivity to domestic livestock and humans should be undertaken.