Browsing by Author "Maia, Carla"
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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.
- Bacterial and protozoal agents of feline vector-borne diseases in domestic and stray cats from southern PortugalPublication . Maia, Carla; Ramos, Claudia; Coimbra, Monica; Bastos, Filipa; Martins, Angela; Pinto, Pedro; Nunes, Monica; Vieira, Maria Luisa; Cardoso, Luis; Campino, LeneaBackground: Feline vector-borne diseases (FVBD) have emerged in recent years, showing a wider geographic distribution and increased global prevalence. In addition to their veterinary importance, domestic cats play a central role in the transmission cycles of some FVBD agents by acting as reservoirs and sentinels, a circumstance that requires a One Health approach. The aim of the present work was to molecularly detect feline vector-borne bacteria and protozoa with veterinary and zoonotic importance, and to assess associated risk factors in cats from southern Portugal. Methods: Six hundred and forty-nine cats (320 domestic and 329 stray), from veterinary medical centres and animal shelters in southern Portugal, were studied. Anaplasma spp./Ehrlichia spp., Babesia spp., Bartonella spp., Borrelia burgdorferi sensu lato, Hepatozoon spp. and Leishmania spp. infections were evaluated by polymerase chain reaction (PCR) in blood samples. Results: One hundred and ninety-four (29.9%) cats were PCR-positive to at least one of the tested genera or complex of FVBD agents. Sixty-four (9.9%) cats were positive to Leishmania spp., 56 (8.6%) to Hepatozoon spp., 43 (6.6%) to Babesia spp., 35 (5.4%) to Anaplasma spp./Ehrlichia spp., 19 (2.9%) to Bartonella spp. and 14 (2.2%) to B. burgdorferi s.l. Thirty-three (5.1%) cats were positive to two (n = 29) or three (n = 4) genera/complex. Babesia vogeli, Bartonella clarridgeiae, Bartonella henselae, Ehrlichia canis, Hepatozoon felis and Leishmania infantum were identified by DNA sequencing. Conclusions: The occurrence of FVBD agents in southern Portugal, some of them with zoonotic character, emphasizes the need to alert the veterinary community, owners and public health authorities for the risk of infection. Control measures should be implemented to prevent the infection of cats, other vertebrate hosts and people.
- Epidemiological and clinical aspects of Cutaneous and Mucosal Leishmaniases in Portugal: retrospective analysis of cases diagnosed in public hospitals and reported in the literature between 2010 and 2020Publication . Rocha, Rafael; Conceição, Cláudia; Gonçalves, Luzia; Carvalho, Ana Cláudia; Maia, André; Martins, André; Carujo, António; Maio, António; Forra, Catarina; Melita, Catarina; Couto, Daniela; Fernandes, Diana; Pereira, Dulce; Leal, Ema; Sarmento, Helena; Sousa, Inês; Gonçalves, Jean-Pierre; Marinho, Joana; Vasconcelos, Joana; Cunha, João; Rodrigues, João; Silva, José Miguel; Caley, Lídia; Malheiro, Luís; Santos, Luís; Garcia, Margarida; Cunha, Maria; Lima, Maria; Andrade, Maria Margarida; Marques, Marta; Alpalhão, Miguel; Silva, Mónica; Ferraz, Rita; Soares, Rui; Fernandes, Salomão; Llobet, Samuel; Cruz, Sofia; Guimarães, Teresa; Branco, Tiago; Nunes, Tomás Robalo; Almeida, Vasco; Maia, CarlaLeishmania infantum, a zoonotic vector-born parasite, is endemic in the Mediterranean region, presenting mostly as visceral (VL), but also as cutaneous (CL) and mucosal leishmaniasis (ML). This study aimed to describe the epidemiological and clinical aspects of the CL and ML cases diagnosed in mainland Portugal between 2010 and 2020. Collaboration was requested from every hospital of the Portuguese National Health System. Cases were screened through a search of diagnostic discharge codes or positive laboratory results for Leishmania infection. Simultaneously, a comprehensive literature search was performed. Descriptive statistics and hypothesis testing were performed using IBM (R) SPSS (R) Statistics. A total of 43 CL and 7 ML cases were identified, with a predominance of autochthonous cases (86%). In CL, immunosuppressed individuals constituted a significant proportion of patients (48%), and in this group, disseminated CL (22%) and simultaneous VL (54%) were common. In autochthonous cases, lesions, mostly papules/nodules (62%), were frequently observed on the head (48%). The approach to treatment was very heterogeneous. ML cases were all autochthonous, were diagnosed primarily in older immunosuppressed individuals, and were generally treated with liposomal amphotericin B. The findings suggest a need for enhanced surveillance and reporting, clinical awareness, and diagnostic capacity of these forms of leishmaniasis to mitigate underdiagnosis and improve patient outcomes. A holistic One Health approach is advocated to address the multifaceted challenges posed by leishmaniases in Portugal and beyond.
- Leishmania infection and host-blood feeding preferences of phlebotomine sandflies and canine leishmaniasis in an endemic European area, the Algarve Region in PortugalPublication . Maia, Carla; Dionisio, Lidia; Afonso, Maria Odete; Neto, Luis; Cristovao, Jose Manuel; Campino, LeneaThe Algarve Region (AR) in southern Portugal, which is an international tourist destination, has been considered an endemic region of zoonotic leishmaniasis caused by Leishmania infantum since the 1980s. In the present study, phlebotomine and canine surveys were conducted to identify sandfly blood meal sources and to update the occurrence of Leishmania infection in vectors and dogs. Four sandfly species were captured: Phlebotomus perniciosus, Phlebotomus ariasi, Phlebotomus sergenti and Sergentomyia minuta. In one P. perniciosus female, L. infantum DNA was detected. Blood meal tests showed that this species had no host preferences and was an opportunistic feeder. An overall canine leishmaniasis (CanL) seroprevalence of 16.06% was found; the seroprevalence was 3.88% in dogs housed in kennels and 40.63% in dogs that attended veterinary clinics. The simultaneous occurrence of dogs and P. perniciosus infected with L. infantum in the AR indicates that the region continues to be an endemic area for CanL. Our results reinforce the need for the systematic spatial distribution of phlebotomine populations and their Leishmania infection rates and the need to simultaneously perform pathogen monitoring in both invertebrate and vertebrate hosts to investigate the transmission, distribution and spreading of Leishmania infection.
- 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 Leishmania infantum, filariae and Wolbachia spp. in dogs from southern PortugalPublication . Maia, Carla; Altet, Laura; Serrano, Lorena; Cristovao, Jose Manuel; Dolores Tabar, Maria; Francino, Olga; Cardoso, Luis; Campino, Lenea; Roura, XavierBackground: Leishmaniosis caused by the protozoan Leishmania infantum and dirofilariosis caused by the nematodes Dirofilaria immitis or Dirofilaria repens are vector-borne zoonoses widely present in the Mediterranean basin. In addition, some studies reported that the endosymbiont Wolbachia spp. play a role in the biology and pathogenesis of filarial parasites. The aim of this work was to evaluate the frequency of mono-and co-infections by L. infantum, filariae and Wolbachia spp. and their association with clinical signs in dogs from the south of Portugal. Leishmanial, filarial and Wolbachia spp. DNA were evaluated by specific real-time polymerase chain reaction (qPCR) assays in blood samples from 230 dogs.Findings: One hundred and thirty-nine (60.4 %) dogs were qPCR-positive for L. infantum and 26 (11.3 %) for filariae (24 for D. immitis only, one D. immitis and for Acanthocheilonema dracunculoides and another one for Acanthocheilonema reconditum only). Wolbachia spp. DNA was amplified from 16 (64.0 %) out of the 25 D. immitis-positive dogs. Nineteen (8.3 %) dogs were co-infected with L. infantum and D. immitis, including the one (0.4 %) A. drancunculoides-positive animal. In dogs without clinical signs consistent with leishmaniosis and/or dirofilariosis, L. infantum prevalence was 69 %, whereas in those dogs with at least one clinical manifestation compatible with any of the two parasitoses prevalence was 42.7 %. Leishmania prevalence was significantly higher in apparently healthy mongrels (77.2 %) and pets (76.9 %) than in defined-breed dogs (including crosses; 58.8 %) and in dogs with an aptitude other than pet (i.e. farm, guard, hunting, shepherd or stray), respectively, whereas in those dogs with at least one clinical sign, the detection of L. infantum DNA was higher in males (53.3 %) and in those dogs not receiving insect repellents (52.8 %).Conclusions: The molecular detection of canine vector-borne disease (CVBD) agents, some of which are zoonotic, reinforces the need to implement efficient prophylactic measures, such as insect repellents and macrocyclic lactones (including compliance to administration), in the geographical areas where these agents are distributed, with the view to prevent infection and disease among mammalian hosts including humans.
- 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.
- Neutralization-based seroprevalence of Toscana virus and sandfly fever Sicilian virus in dogs and cats from PortugalPublication . Alwassouf, Sulaf; Maia, Carla; Ayhan, Nazli; Coimbra, Mónica; Cristovão, José Manuel; Richet, Herve; Bichaud, Laurence; Campino, Lenea; Charrel, Remi N.Sandfly-borne phleboviruses are endemic in the Mediterranean basin. However, levels of exposure of human and animal populations are inadequately researched. Toscana virus (TOSV) is present in Portugal where it causes human infection and disease; in contrast there are few data for sandfly fever Sicilian virus (SFSV) which has neither been isolated nor detected by molecular tests and for which there are only limited serological data. The sera collected from 1160 dogs and 189 cats in southern Portugal were tested for the presence of neutralizing antibodies against TOSV and SFSV, two viruses recognized as distinct serocomplexes in the Mediterranean region. Our data showed (i) seropositivity to TOSV and SFSV in dogs at a rate of 6.8 and 50.8 %, respectively, and (ii) that 3.7 % of cats were seropositive for TOSV. TOSV findings are in line with previous results obtained with less stringent serological assays. Our results for SFSV in dogs clearly indicate that the virus is circulating widely and that humans may be exposed to infection via the dogs. Although the presence of SFSV was suggested by haemagglutination inhibition in 4/1690 human sera in 1974, this is the first time, as far as we know, that SFSV has been shown to circulate so widely in dogs in Portugal. Future studies should be directed at isolating strains of SFSV in Portugal from dogs, humans and sandflies collected in high prevalence regions. As dogs appear to be good sentinels for SFSV, their role as a possible reservoir in the natural cycle should also be considered.
- Prevalence and correlates of antibodies to Neospora caninum in dogs in PortugalPublication . Maia, Carla; Cortes, Helder; Brancal, Hugo; Lopes, Ana Patricia; Pimenta, Paulo; Campino, Lenea; Cardoso, LuisNeosporosis, caused by Neospora caninum, is an important cause of abortion in cattle and of neurological disease in dogs. This study investigated the prevalence and correlates of antibodies to N. caninum in 441 dogs from the five regions of mainland Portugal. A commercial competitive enzyme-linked immunosorbent assay (cELISA) was used and specific antibodies were detected in 35 (7.9%) dogs. Seroprevalence levels were significantly different among some of the studied regions, as well as between stray dogs (13.6%) and hunting dogs (1.7%). The difference between seropositivity in dogs presenting musculoskeletal or neurological signs (21.4%) and that in animals without clinical signs compatible with neosporosis (5.6%) was close to statistical significance. This is the first report on the seroprevalence of N. caninum in dogs in Portugal. Neosporosis should be considered in the differential diagnosis of neurological disorders of dogs.
- Serological investigation of Leishmania infantum, Dirofilaria immitis and Angiostrongylus vasorum in dogs from southern PortugalPublication . Maia, Carla; Coimbra, Monica; Ramos, Claudia; Cristovao, Jose Manuel; Cardoso, Luis; Campino, LeneaBackground: Leishmaniosis, dirofilariosis and angiostrongylosis are parasitic diseases of established importance in dogs worldwide. The aim of the present report was to determine all together levels of infection with or exposure to Leishmania infantum, Dirofilaria immitis and Angiostrongylus vasorum in dogs from the Algarve region, in southern Portugal. Findings: Serum samples were obtained from a total of 170 apparently healthy dogs. Antibodies to L. infantum were detected by the direct agglutination test; and antigens of D. immitis and of A. vasorum by an enzyme-linked immunosorbent assay and an immunochromatography test, respectively. Antibodies to L. infantum were found in 18.2% (31/170) of dogs, while D. immitis antigen was detected in 9.4% (16/170). None of 120 dogs tested out of the 170 under investigation had evidence of infection with A. vasorum. D. immitis infection was more prevalent in dogs aged 84-204 months than in those younger than 12 months. Three dogs were positive to both L. infantum and D. immitis. Conclusions: Dogs living in southern Portugal are at risk of leishmaniosis and dirofilariosis. This scenario should make the veterinary community, local dog owners and also tourists from non-endemic countries coming on vacation with their pets aware of the need of effective prophylactic measures, in order to protect animals and public health.