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Seroprevalence of vector-borne pathogens and molecular detection of Borrelia afzelii in military dogs from Portugal

Publication . Alho, Ana Margarida; Pita, Joana; Amaro, Ana; Amaro, Fátima; Schnyder, Manuela; Grimm, Felix; Custódio, Ana Cristina; Cardoso, Luís; Deplazes, Peter; de Carvalho, Luís Madeira

Canine vector-borne diseases (CVBDs) are increasingly being reported worldwide and represent a serious threat to both animal and public health. Military dogs may constitute a risk group for the agents causing these diseases, as they frequently work outdoors in different areas and are thus exposed to vector arthropods. In order to assess the risk of exposure of this type of dogs, a serological and molecular survey was conducted in military working dogs in Portugal. One hundred apparently healthy dogs were surveyed. Serum samples were tested for antigens of Angiostrongylus vasorum and Dirofilaria immitis; and for antibodies to A. vasorum, Anaplasma spp., Babesia spp., Ehrlichia canis, Leishmania infantum, Rickettsia spp. and Toscana virus. Serum was tested by polymerase chain reaction for Borrelia burgdorferi (sensu lato), with sequencing of the DNA products.

2016-05-10Journal article

Open access

Phenotypic and Molecular Characterization of Salmonella 1,4,[5],12:i:- R-Type ASSuT Isolates from Humans, Animals, and Environment in Portugal, 2006-2011

Publication . Seixas, Rui; Santos, Tânia Raquel; Machado, Jorge; Tavares, Luís; Bernardo, Fernando; Semedo-Lemsaddek, Teresa; Oliveira, Manuela

The increase in prevalence of Salmonella 1,4,[5],12:i:- related infections over the last few years has been considered a public health issue in many European countries, especially as this serovar may be associated with tetraresistance to ampicillin, streptomycin, sulfonamides, and tetracyclines (R-type ASSuT). Salmonella 1,4,[5],12:i:- isolates (n = 187) obtained by the Portuguese National Laboratory from different sources, including human clinical cases (n = 170), veterinary (n = 10), environmental (n = 6), and food samples (n = 1), were collected from 15 districts between 2006 and 2011. All isolates were serotyped using the slide agglutination method and results were confirmed by multiplex PCR for the monophasic variant. From the confirmed Salmonella 1,4,[5],12:i:-, R-type ASSuT isolates were selected by disc diffusion and minimal inhibitory concentration (MIC) determination for further characterization by pulsed-field gel electrophoresis restriction with XbaI, virulence genes determination by PCR, additional antimicrobial resistance profiling by disc diffusion, and epidemiological distribution evaluation. Out of the 187 serotyped isolates, 133 were confirmed as Salmonella 1,4,[5],12:i:- with a R-type ASSuT occurrence of 61.7%. Distribution among Portuguese districts showed a higher percentage of reported cases in coastal areas, in particular, in Porto (24.8%), Setúbal (13.5%), and Aveiro (12.8%), probably due to the higher population density. Clonality analysis revealed a high diversity of pulsotypes with the majority of human salmonellosis cases being attributed to sporadic events. All isolates harbored 14 out of the 18 virulence genes evaluated and 87.8% of the isolates showed all the resistance genes frequently associated with the European clone, blaTEM(+)sul2(+)straA-straB(+)tetB(+). This study shows that Salmonella 1,4,[5],12:i:- resistant isolates are widely distributed in Portugal. This may be related to a selective advantage offered by R-type ASSuT profile, the presence of multiple virulent features, including the ability to form biofilms, which along with a high diversity of pulsotypes may be responsible for the dissemination through the country.

2016-11Journal article

Restricted access

Demographic characterization and spatial cluster analysis of human Salmonella 1,4,[5],12:i:- infections in Portugal: A 10 year study

Publication . Seixas, R.; Nunes, T.; Machado, J.; Tavares, L.; Owen, S.P.; Bernardo, F.; Oliveira, M.

Salmonella 1,4,[5],12:i:- is presently considered one of the major serovars responsible for human salmonellosis worldwide. Due to its recent emergence, studies assessing the demographic characterization and spatial epidemiology of salmonellosis 1,4,[5],12:i:- at local- or country-level are lacking. In this study, a analysis was conducted over a 10year period, from 2000 to the first quarter of 2011 at the Portuguese National Laboratory in Portugal mainland, with a total of 215 Salmonella 1,4,[5],12:i:- serotyped isolates obtained from human infections by a passive surveillance system. Data regarding source, year and month of sampling, gender, age, district and municipality of the patients were registered. Descriptive statistical analysis and a spatial scan statistic combined with a geographic information system were employed to characterize the epidemiology and identify spatial clusters. Results showed that most districts have reports of Salmonella 1,4,[5],12:i:-, with a higher number of cases at the Portuguese coastland, including districts like Porto (n=60, 27.9%), Lisboa (n=29, 13.5%) and Aveiro (n=28, 13.0%). An increased incidence was observed in the period from 2004 to 2011 and most infections occurred during May and October. Spatial analysis revealed 4 clusters of higher than expected infection rates. Three were located in the north of Portugal, including two at the coastland (Cluster 1 [RR=3.58, p≤0.001] and 4 [RR=10.42 p≤0.230]), and one at the countryside (Cluster 3 [RR=17.76, p≤0.001]). A larger cluster was detected involving the center and south of Portugal (Cluster 2 [RR=4.85, p≤0.001]). The present study was elaborated with data provided by a passive surveillance system, which may originate an underestimation of disease burden. However, this is the first report describing the incidence and the distribution of areas with higher risk of infection in Portugal, revealing that Salmonella 1,4,[5],12:i:- displayed a significant geographic clustering and these areas should be further evaluated to identify risk factors in order to establish prevention programs.

2018-03Journal article

Open access