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Research Project
DEVELOPMENT OF AN EXPERIMENTAL MODEL OF LEISHMANIA INFANTUM FOR THE EVALUATION OF VACCINE CANDIDATES
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The recombinant protein rSP03B is a valid antigen for screening dog exposure to Phlebotomus perniciosus across foci of canine leishmaniasis
Publication . Kostalova, T.; Lestinova, T.; Maia, C.; Sumova, P.; Vlkova, M.; Willen, L.; Polanska, N.; Fiorentino, E.; Scalone, A.; Oliva, G.; Veronesi, F.; Cristovao, J. M.; Courtenay, O.; Campino, Lenea; Gradoni, L.; Gramiccia, M.; Volf, P.
The frequency of sandfly-host contacts can be measured by host antibody levels against sandfly salivary proteins. Recombinant salivary proteins are suggested to represent a valid replacement for salivary gland homogenate (SGH); however, it is necessary to prove that such antigens are recognized by antibodies against various populations of the same species. Phlebotomus perniciosus (Diptera: Psychodidae) is the main vector of Leishmania infantum (Trypanosomatida: Trypanosomatidae) in southwest Europe and is widespread from Portugal to Italy. In this study, sera were sampled from naturally exposed dogs from distant regions, including Campania (southern Italy), Umbria (central Italy) and the metropolitan Lisbon region (Portugal), where P. perniciosus is the unique or principal vector species. Sera were screened for anti-P. perniciosus antibodies using SGH and 43-kDa yellow-related recombinant protein (rSP03B). Arobust correlation between antibodies recognizing SGH and rSP03B was detected in all regions, suggesting substantial antigenic cross-reactivity among different P. perniciosus populations. No significant differences in this relationship were detected between regions. Moreover, rSP03B and the native yellow-related protein were shown to share similar antigenic epitopes, as canine immunoglobulin G (IgG) binding to the native protein was inhibited by pre-incubation with the recombinant form. These findings suggest that rSP03B should be regarded as a universal marker of sandfly exposure throughout the geographical distribution of P. perniciosus.
Molecular detection of Leishmania DNA and identification of blood meals in wild caught phlebotomine sand flies (Diptera: Psychodidae) from southern Portugal
Publication . Maia, Carla; Parreira, Ricardo; Cristovao, Jose Manuel; Freitas, Ferdinando Bernardino; Afonso, Maria Odete; Campino, Lenea
Background: 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.
Prevalence and correlates of antibodies to Neospora caninum in dogs in Portugal
Publication . Maia, Carla; Cortes, Helder; Brancal, Hugo; Lopes, Ana Patricia; Pimenta, Paulo; Campino, Lenea; Cardoso, Luis
Neosporosis, 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.
Leishmania infection and host-blood feeding preferences of phlebotomine sandflies and canine leishmaniasis in an endemic European area, the Algarve Region in Portugal
Publication . Maia, Carla; Dionisio, Lidia; Afonso, Maria Odete; Neto, Luis; Cristovao, Jose Manuel; Campino, Lenea
The 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.
Emergence of Thelazia callipaeda Infection in dogs and cats from east-central Portugal
Publication . Maia, C.; Catarino, A. L.; Almeida, B.; Ramos, C.; Campino, Lenea; Cardoso, L.
The eyeworm Thelazia callipaeda (Spirurida, Thelaziidae) infects domestic animals, wildlife and human beings, and is considered an emerging pathogen in Europe. This study aimed at investigating the prevalence and risk factors of T.callipaeda infection in dogs and cats from east-central Portugal, a region where the parasite was previously detected in two red foxes (Vulpes vulpes). Thelazia callipaeda was found in 22 (3.8%) of 586 dogs and in four (23.5%) of 17 cats. A total of 178 adult worms (71.9% of females and 28.1% of males) were collected from the conjunctiva of the infected dogs. The number of worms collected per dog ranged from 1 to 35 (average +/- standard deviation: 8.08 +/- 9.49), with four dogs (18.2%) harbouring only a single parasite. Worms were gathered from dogs throughout all months of the year. A total of 17 adult worms (64.7% of females and 35.3% of males) were obtained from cats. The number of worms per cat ranged from 1 to 14 (4.3 +/- 6.5), with three cats (75.0%) having a single parasite. Eyeworm infection was statistically more prevalent in pastoral and farm dogs, in those dogs with contact with other animals and in dogs with ocular manifestations. T. callipaeda is endemic in the east-central part of Portugal, reportedly infecting domestic (dogs and cats) and wild carnivores (red foxes) and evidencing a southerly dissemination. Future investigations should be focused on determining the local distribution and density of the insect vector (Phortica variegata) in this geographical area. This emergent zoonosis should be included by veterinarians, physicians and ophthalmologists in the differential diagnosis of ocular manifestations in their patients, particularly in areas where T. callipaeda is endemic.
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Fundação para a Ciência e a Tecnologia
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Funding Award Number
SFRH/BPD/44082/2008