2010) and serological methods (Schnyder et al. for in dogs from Portugal. The endemic occurrence of in dogs from different geographical areas of Portugal is usually therefore confirmed. first stage larvae (L1), with the characteristic kinked tail, dorsal spine and notch feature (Guilhon and Mouse monoclonal to HSP60 Cens 1973). FLOTAC, an improved flotation-based coproscopic method, also allows for the visualisation of L1 in faecal samples, with a good sensitivity (Schnyder et al. 2011a). However, due to prepatency, intermittent larval excretion and the possible occurrence of mixed lungworm infections, copromicroscopic techniques have limitations concerning sensitivity and specificity. Besides, by the time dogs start to be positive in Baermann or FLOTAC, damage to the lung parenchyma is already present, and recovery is usually more difficult (Guilhon and Cens 1969; Neff 1971; SB 218078 Dennler et al. 2011). Newly developed diagnostic techniques, such as PCR (Jefferies et al. 2009; Al-Sabi et al. 2010) and serological methods (Schnyder et al. 2011b; Schucan et al. 2012), have been designed to detect infected animals. Serological methods were shown to be highly suitable for both individual and massive screening of doggie populations. In fact, serologies require single serum samples instead of repeated faecal samples and allows for rapid detection of infection, shortly before or contemporaneously with patency (Schnyder et al. 2015b). Regarding the geographical distribution of has a very heterogeneous distribution with reports suggesting the presence of endemic hotspots in many areas, namely in Croatia (Rajkovic-Janje et al. 2002), Italy (Della Santa et al. 2002; Guardone et al. 2013), Switzerland (Staebler et al. 2005), Germany (Staebler et al. 2005; Barutzki and Schaper 2009), Spain (Segovia et al. 2004; Ma?as et al. 2005), Greece (Papazahariadou et al. 2007), Poland (Demiaszkiewicz et al. 2014), Slovakia (Miterpakova et al. 2014), Hungary (Schnyder et al. 2015a) as well as others. Several hypotheses have been raised to explain this possible expansion, such as increased movements of pet dogs and increased fox populations even in urban areas, suggesting that new areas are open to colonisation (Morgan et al. 2009). In Portugal, knowledge concerning the current situation of contamination in domestic and wild canids is usually poor. No studies conducted so far showed positive results, and no surveillance mechanisms are in place to assess its prevalence or geographical range. was first identified during the necropsy of one of 306 red foxes (was sporadically recognized in domestic dogs, with three positive cases diagnosed in the last few years in the Lisbon area (Madeira de Carvalho et al. 2009, 2013; Nabais et al. 2014). A serological study using a commercial antigen kit (Angio DetectTM Test, IDEXX Laboratories) tested negative around the 120 surveyed dogs from your Algarve region (Maia et al. 2015). The present serological study aimed to increase the knowledge about the occurrence and geographical dispersion of infections in Portugal. Material and methods A total of 906 shelter SB 218078 dogs randomly distributed from north to south of mainland Portugal were studied. All animals were stray dogs, and no information was available regarding previous preventive treatments. Blood samples (2C3?ml) were collected from your cephalic vein, and serum was separated by centrifugation and stored at ?20?C until use. Sera were tested at the Institute of Parasitology, Vetsuisse Faculty, University or college of Zurich, Switzerland, for the presence of circulating antigens using monoclonal and polyclonal antibodies in a sandwich ELISA, with a sensitivity of 95.7?% and a specificity of 94.0?%, as previously explained (Schnyder et al. 2011b). Additionally, a sandwich ELISA (sensitivity 81.0?%, specificity 98.8?%) using adult somatic antigen purified by monoclonal antibodies (mAb Av 5/5) was utilized for specific antibody detection (Schucan et al. 2012). Test thresholds (Schnyder et al. 2013a) were regionally decided with 300 randomly determined samples based on the mean value of optical density (A405 nm) plus three standard deviations. All test runs included a background control, a conjugate control, three positive control sera from SB 218078 three experimentally infected dogs and two unfavorable control sera from uninfected dogs. The collected data were analysed using a geographical information system (GIS) program (RegioGraph 10, GfK GeoMarketing, Bruchsal, Germany) to visualise the regional distribution of collected and analysed serum samples and antigen- and/or antibody-positive samples. The locations of positive samples were displayed on maps with administrative and postcode boundaries based on the Portuguese four-digit postcode as points of reference. Excel 2007 for Windows (Microsoft Corporation, Redmond, USA) was used to calculate the prevalence values and their 95?% confidence interval (CI). Results A total of 0.66?% of the dogs (antigen and antibody detection, and 12 dogs (1.32?%, 95?% CI 0.68C2.30) were.