First report of apparent praziquantel resistance in Dipylidium caninum in Europe.

Dipylidium caninum is a common tapeworm of dogs. Two cases of praziquantel resistance have been described in D. caninum in the United States. No further reports have been published to the authors' knowledge. Here, the case of a dog imported to Switzerland from Spain with a history of chronic excretion of tapeworm proglottids and unresponsiveness to praziquantel treatments is reported. Clinical signs were mild (restlessness, tenesmus, anal pruritus, squashy feces) and flea infestation could be ruled out. Infection with D. caninum was confirmed through morphological and genetic parasite identification. Different subsequently applied anthelmintic compounds and protocols, including epsiprantel, did not confer the desired effects. Proglottid shedding only stopped after oral mebendazole administration of 86.2 mg kg−1 body weight for 5 consecutive days. Clinical signs resolved and the dog remained coproscopically negative during a follow-up period of 10 months after the last treatment. This case represents the first reported apparent praziquantel and epsiprantel resistance in D. caninum in Europe. Treatment was extremely challenging especially due to the limited availability of efficacious alternative compounds.

A pilot study for the isolation of Eimeria spp. oocysts from environmental straw samples in comparison with individual faecal examination of fattening calves.

The diagnosis of eimeriosis in calves mainly relies on the presence of diarrhoea and the excretion of Eimeria oocysts in the faeces. Restraining the animals to collect rectal samples for diagnostic purposes is stressful and time-consuming. The aim of this study was to evaluate a method for the quantification of oocysts in environmental barn straw samples. To investigate the recovery rate of the method, straw and Eimeria negative faeces were spiked with Eimeria oocysts in plastic bags and mixed with water and 0.05% Tween 20 (v/v); the liquids were filtered twice through sieves (mesh size 300 and 52 µm), centrifuged and the number of oocysts in the sediment determined using a McMaster counting chamber. A recovery rate of 52.4% (95% confidence interval: 48.2-56.5%) was obtained. In the following, field straw (n = 156) and individual faecal samples (n = 195, also analysed by McMaster counting chambers) were collected on four different farms. Eimeria oocysts were present on all farms in faecal (84/195, 43.1%) and straw samples (119/156, 76.3%). In 37 (23.7%) straw samples, sporulated oocysts were observed, with a sporulation rate ranging from 0 to 40%. Despite high variability between farms and examination days, mean numbers of oocysts in the straw positively correlated with mean numbers of oocysts excreted in the faeces (ρSpearman = 0.60). The examination of environmental straw samples may represent an easy-to-perform, non-invasive, inexpensive preliminary diagnostic approach for surveillance of eimeriosis at group level, having the potential to assess the infection pressure.

Estimated specific antibody-based true sero-prevalences of canine filariosis in dogs in Central Europe and the UK.

Dirofilariosis is a vector-borne disease mainly caused by Dirofilaria immitis and Dirofilaria repens. In contrast to the known endemicity of dirofilariosis in southern and south-eastern Europe, information on the distribution of D. repens in Central-Europe is fragmentary. We tested 8877 serum samples from dogs from Austria, Denmark, Germany, Italy, Lithuania, Poland, Switzerland and the UK using an ELISA detecting filarial-specific antibodies, hypothesising higher occurrence of D. repens. Based on two overlapping frequency distributions, presumed negative samples had a mean optical density (OD) value of 0.097, representing 97.45% of all samples. Presumed positive samples, representing 2.55% of all sera, had a mean OD value of 0.287. Test prevalence based on the calculated cut-off was 3.51% for all sera (4.36% for Austria, 1.94% for Denmark, 1.39% for Germany, 3.37% for Italy, 6.90% for Lithuania, 6.99% for Poland, 0.77% for Switzerland and 0.0% for the UK, respectively). The bimodal distribution, representing overlapping distributions of OD values from positive and negative dogs, enabled the assignment of a probability of true infection status to each dog. Mean probabilities of true infection status across groups, based on the postal codes of origin, allowed us to estimate and map true prevalences. For all countries, except the UK, the true prevalence was lower than the test prevalence. The large number of serum samples and the use of a non-gold standard analytical method allowed us to create a more realistic picture of the distribution of D. repens in Central Europe and the UK.

Genome sequence of the cardiopulmonary canid nematode Angiostrongylus vasorum reveals species-specific genes with potential involvement in coagulopathy.

Angiostrongylus vasorum is an emerging parasitic nematode of canids and causes respiratory distress, bleeding, and other signs in dogs. Despite its clinical importance, the molecular toolbox allowing the study of the parasite is incomplete. To address this gap, we have sequenced its nuclear genome using Oxford nanopore sequencing, polished with Illumina reads. The size of the final genome is 280 Mb comprising 468 contigs, with an N50 value of 1.68 Mb and a BUSCO score of 93.5%. Ninety-three percent of 13,766 predicted genes were assigned to putative functions. Three folate carriers were found exclusively in A. vasorum, with potential involvement in host coagulopathy. A screen for previously identified vaccine candidates, the aminopeptidase H11 and the somatic protein rHc23, revealed homologs in A. vasorum. The genome sequence will provide a foundation for the development of new tools against canine angiostrongylosis, supporting the identification of potential drug and vaccine targets.

Conquering Switzerland: the emergence of Angiostrongylus vasorum in foxes over three decades and its rapid regional increase in prevalence contrast with the stable occurrence of lungworms.

Angiostrongylus vasorum, Crenosoma vulpis and Capillaria aerophila are the most common lungworms of domestic and wild canids. We investigated the short- and long-term lungworm prevalence changes in the Swiss fox population with a focus on A. vasorum. Between 2012 and 2017, lungs and hearts of 533 foxes from north-eastern Switzerland were necropsied and blood samples tested for circulating A. vasorum antigen. Angiostrongylus vasorum prevalence increased steadily from 21.5% in 2012 to 81.8% in 2017. In contrast, C. aerophila and C. vulpis prevalences fluctuated between 41.8 and 74.7%, and 3.6 and 14.9%, respectively. Based on 3955 blood samples collected between 1986 and 2017 from three geographic areas and during four time periods, antigen seropositivity increased from 2.4 to 62.0%. In north-eastern Switzerland, seropositivity was initially low (1.9 and 1.7% in the first two time periods) but increased in the following two decades to 22.2 and 62.0%, respectively. Our findings depict the spectacular expansion of A. vasorum in the past three decades. Regionally, the prevalence in foxes increased 4-fold within 6 years in some regions. This underpins the important role of foxes as reservoir hosts, likely explaining the increasing number of cases of canine angiostrongylosis in Switzerland. Our findings are representative of central Europe and may help anticipating future developments in areas where A. vasorum is present but (still) infrequent.

Feline lungworms in Greece: copromicroscopic, molecular and serological study.

Aelurostrongylus abstrusus (Nematoda, Metastrongyloidea) causes verminous pneumonia in cats worldwide. This study evaluated the seroprevalence of A. abstrusus antibodies in 220 stray and free-roaming cats from insular (Mykonos, Crete, Skopelos) and continental (Thessaloniki, Attica) Greece. The results were compared with morphological and biomolecular identification of first-stage larvae (L1) in faeces. Positive cats were observed in all 5 areas: 13/97 (13.4%), 1/32 (3.1%), 7/26 (26.9%), 3/18 (16.7%) and 5/47 (10.6%) cats tested positive for A. abstrusus L1 by Baermann examination, and 33/97 (34.0%), 7/32 (21.9%), 6/26 (23.1%), 3/18 (16.7%) and 11/47 (23.4%) were seropositive, in Mykonos, Crete, Skopelos, Thessaloniki and Attica, respectively. Troglostrongylus brevior L1 were found in 12/97 (12.4%), 3/26 (11.5%) and 2/47 (4.3%) cats from Mykonos, Skopelos and Attica respectively. Six of the 220 cats (2.7%), i.e. 4/97 (4.1%) from Mykonos and 2/26 (7.7%) from Skopelos, shed L1 of both A. abstrusus and T. brevior. Sixty samples were ELISA-positive (27.3%, 95% CI: 21.5-33.7%), of which 21 (35%) tested copromicroscopically positive (19 monospecific infections and 2 mixed with Troglostrongylus brevior), and 5 were positive for T. brevior L1 only. Among seronegative cats (n = 140), L1 of A. abstrusus were additionally detected in 8 (5.7% out of 140) cats (i.e. 4 monospecific infections and 4 mixed with T. brevior), and in 6 (4.3% out of 140) cats, L1 of T. brevior as monospecific infection were detected. This study confirms the presence of lungworms in Greece and suggests that the number of cats infected with/exposed to metastrongylids is higher than detected by faecal examinations.

A grid-cell based fecal sampling scheme reveals: land-use and altitude affect prevalence rates of Angiostrongylus vasorum and other parasites of red foxes (Vulpes vulpes).

In view of the role of foxes as a reservoir for Angiostrongylus vasorum, a nematode of the heart and lungs of dogs and foxes, its occurrence across Switzerland was investigated in foxes applying a standardized sampling scheme for fox fecal samples. In 72 study areas, which consisted of three 1-km2 grid cells, a total of 1481 samples were collected by walking transects following linear features in the terrain, and analyzed by a flotation-sieving method. The overall prevalence rate of A. vasorum in fecal samples was 8.8% (95% confidence interval, CI 7.4-10.3%), being significantly higher in the Swiss Plateau (11.4%, CI. 9.4-13.8%) compared with other bioregions, and more prevalent in areas with less than 50% of cultivated land (p = 0.043). Prevalence rates increased with decreasing altitudes being significantly higher below 400 m above sea level (20.6%, 95% CI 15.2-26.9%), while all samples collected above 900 m asl were negative. Eggs of Toxocara sp. (12.1%), Taeniidae (10.5%), Capillaria spp. (8.3%), Trichuris vulpis (5.5%), hookworms (5.3%), Toxascaris leonina (1.3%) and Strongyloides sp. (0.4%) were furthermore identified. Taeniid eggs were positively and Capillaria spp. negatively associated with the amount of cultivated land. The prevalence rates based on our fecal analyses were generally lower compared with previous studies from Switzerland which were based on fox necropsies. However, the grid cell-based sampling scheme with replicable transects for fox feces collection proved to be a suitable, non-invasive method for parasitic large-scale surveys providing information on the relative infectious pressure for the local dog population.

Serological survey and risk factors of Aelurostrongylus abstrusus infection among owned cats in Italy.

Feline lungworms affect the respiratory tract of domestic cats causing respiratory conditions of various degrees. In this study, we investigated the exposure of cats to feline lungworm infections by detecting antibodies in a large population of animals from several regions of Italy. Sera of 1087 domestic cats living in regions of the north (n = 700), the centre (n = 227) and the south (n = 160) of Italy were examined by a newly developed indirect ELISA conceived for detection of antibodies against the most frequently occurring feline lungworm Aelurostrongylus abstrusus. Individual cat data (i.e., age, sex, neutering status and provenience) were analysed as potential risk factors for exposure to lungworm infections. Samples were additionally screened for feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) proviral DNAs. Overall, 9% (98/1087; 95% confidence interval (CI) 7.4-10.9%) of the animals tested seropositive to lungworm antibodies. Positive cats were identified in the north (7.1%; CI 5.5-9.3%), in the centre (5.3%; CI 3.0-9.0%) and in the South (22.5%; CI 16.7-29.6%), with more seropositive animals in the latter area (p < 0.05). The risk of lungworm infection in cats was significantly associated with age less than 6 months (i.e. 24.4%, p < 0.05) and FIV infection (p < 0.05). This large-scale serological survey confirms the exposure of cats to lungworm infections in Italy and that serological tests can be used to assess the distribution of lungworm infections in large populations of animals.

Seroprevalence of circulating Angiostrongylus vasorum antigen and parasite-specific antibodies in dogs from Portugal.

Angiostrongylus vasorum is a nematode that lives in the pulmonary arteries and right cardiac ventricle of domestic dogs and wild canids. It is increasingly being reported in several European countries and North America. This parasite induces inflammatory verminous pneumonia, causing severe respiratory disease in dogs. In some instances, coagulopathies, neurological signs and even death may occur. Scant data are available regarding the occurrence of A. vasorum in Portugal. Therefore, sera of 906 shelter dogs from North to South mainland Portugal were collected. ELISAs to detect A. vasorum circulating antigen and specific antibodies against this parasite were performed. A total of six dogs [0.66 %, 95 % confidence intervals (CI) 0.24-1.43] were positive for both A. vasorum antigen and antibody detection, indicating an active infection, and 12 dogs (1.32 %, CI 0.68-2.30) were A. vasorum antibody-positive only. Regions with antigen- and antibody-positive animals overlapped and were distributed over nearly all sampled areas in the country. This is the first large-scale ELISA-based serological survey for A. vasorum in dogs from Portugal. The endemic occurrence of A. vasorum in dogs from different geographical areas of Portugal is therefore confirmed.

Combined Serological Detection of Circulating Angiostrongylus vasorum Antigen and Parasite-specific Antibodies in Dogs from Hungary.

The occurrence of the nematode Angiostrongylus vasorum, also known as the French heartworm, is increasingly being reported from various European countries. The adults of this parasite species live in the pulmonary arteries and right cardiac ventricle of wild canids and domestic dogs. Larval stages and eggs in the lungs induce inflammatory verminous pneumonia, causing severe respiratory disease in dogs. Furthermore, haematological and neurological signs and even death may occur. In Hungary, A. vasorum has been identified in red foxes, golden jackals and in two dogs and some slugs. In this first large-scale survey, 1247 sera from pet dogs were collected and tested by an ELISA for the detection of circulating antigen of A. vasorum and by a separate ELISA to detect specific antibodies against the parasite. A total of 1.36% (n = 17, 95 % confidence intervals, CI: 0.80 - 2.17 %) of the animals were positive in both ELISAs, while 1.76 % (n = 22, CI: 1.11 - 2.66 %) of the tested dogs were antigen-positive only and 2.73 % (n = 34, CI: 1.90 - 3.79 %) were positive for specific antibodies only. Regions with antigen- and antibody-positive animals overlapped and were distributed over nearly the whole sampled areas of the country. A considerable number of cases was observed in Budapest and also in the southern part of the country bordering Croatia, while in the most eastern part bordering Ukraine no positive samples were detected. These results confirm the endemic occurrence of A. vasorum in dogs originating from different parts of Hungary and the significant advantages of A. vasorum serology in epidemiological studies.

Current Surveys of the Seroprevalence of Borrelia burgdorferi, Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, Babesia canis, Angiostrongylus vasorum and Dirofilaria immitis in Dogs in Bulgaria.

Canine vector-borne diseases (CVBDs) have increasingly become a focus of interest in recent years. Some of the CVBDs are zoonotic and may therefore also represent a risk for the human population. Different factors are in discussion to explain the expansion of vectors and pathogens into formerly unaffected areas. Knowledge of the prevalence and distribution of CVBDs in Bulgaria is scant overall and most data rely on single case descriptions. The aim of the present study was to determine the seroprevalence of important CVBDs in 167 dogs from central-southern Bulgaria (Stara Zagora), with special emphasis on hitherto uninvestigated babesiosis and angiostrongylosis, on poorly investigated Lyme borreliosis and canine granulocytic anaplasmosis, and on the potentially zoonotic dirofilariosis and leishmaniosis. Relatively high prevalence rates were documented for anti-Babesia canis antibodies, Dirofilaria immitis antigen (16.2 %; 27/167 each), anti-Ehrlichia canis (21 %; 35/167) and anti-Anaplasma phagocytophilum antibodies (30.5 - 46.1 %; 51 - 77/167), while Borrelia burgdorferi seroprevalence was low (2.4 %; 4/167). All samples were negative for Leishmania infantum antibodies and Angiostrongylus vasorum antigen and antibodies. In total, 64.7 % (108/167) of the samples indicated infection or exposure to at least one agent and a high proportion of dual infections (39.8 %; 43/108) was demonstrated. Multiple infections with up to four different organisms were also detected. Our data underline the importance of CVBDs and especially of co-infections which could influence the clinical outcome in dogs.

Efficacy of Emodepside/Praziquantel Spot-on (Profender®) against adult Aelurostrongylus abstrusus Nematodes in Experimentally Infected Cats.

The adulticidal efficacy of a topical combination of emodepside 2.1 % (w/v) plus praziquantel 8.6 % (w/v) (Profender® spot-on for cats, Bayer) against adult Aelurostrongylus abstrusus nematodes was evaluated in two randomised, placebo-controlled laboratory efficacy studies. Each study involved 16 cats experimentally inoculated with L3 (800 and 600 each in studies no. 1 and 2, respectively) and randomised into two study groups of 8 cats each after onset of patency. While cats in the treatment group in study no. 1 received a single spot-on application at the minimum therapeutic dose (3 mg/kg emodepside and 12 mg/kg praziquantel), cats in study no. 2 were treated twice with an interval of 14 days. The faecal output of first stage larvae was monitored throughout the study. Necropsy was conducted 4 or 5 weeks after the (first) treatment and the worm counts were used for efficacy calculations. The control groups showed a geometric mean of the total worm count (live and dead worms) of 28.8 (study no. 1) and 17.6 (study no. 2), respectively. All control animals were infected. While the single treatment in study no. 1 resulted in a reduction of the total worm burden by 73.0 % (p = 0.0070), the treatment protocol in study no. 2 was 99.2 % effective (p = 0.0035). Based on live worm counts, the efficacy in study no. 2 was 100 % (p = 0.0030). It is concluded that two applications of Profender® spot-on given two weeks apart represent a safe and highly efficacious treatment regime against feline aelurostrongylosis.

Stability of the southern European border of Echinococcus multilocularis in the Alps: evidence that Microtus arvalis is a limiting factor.

SUMMARY The known range of the zoonotic fox tapeworm Echinococcus multilocularis has expanded since the 1990s, and today this parasite is recorded in higher abundances throughout large parts of Europe. This phenomenon is mostly attributed to the increasing European fox populations and their invasion of urban habitats. However, these factors alone are insufficient to explain the heterogeneous distribution of the parasite in Europe. Here, we analysed the spatial interrelationship of E. multilocularis with the known distribution of seven vole species in Ticino, southern Switzerland. Among 404 necropsied foxes (1990-2006) and 79 fox faecal samples (2010-2012), E. multilocularis was consistently found in the north of the investigated area. No expansion of this endemic focus was recorded during the 22 years of the study period. This stable endemic focus is coincident with the known distribution of the vole species Microtus arvalis but not, or only partly, with the distribution of the other autochthonous vole species. Our results give evidence that this vole species plays a crucial role in the maintenance of the parasite's life cycle and that its absence could be a limiting factor for the spread of E. multilocularis in this region.

Control of companion animal parasites and impact on One Health.

The last decades have witnessed an increase in the global population and movements of companion animals, contributing to changes in density and distribution of pet parasites. Control of companion animal parasites (CAPs) becomes increasingly relevant because of the intensifying human-animal bond. Parasites impact on the health of humans and their pets, but also of wildlife and the environment. We conducted a qualitative review on the current advancements, gaps and priorities for the monitoring and treatment of CAPs with a focus on securing public health. There is a need to raise awareness, coordinate global surveillance schemes and better quantify the impact of companion animal parasites on One Health.

Extended-spectrum ß-lactamase (ESBL)- and carbapenemase-producing Enterobacterales isolated from fresh herbs and salads at retail level in Switzerland.

Fresh produce is usually consumed raw or minimally processed, making it a potential vehicle for transmission of antimicrobial-resistant (AMR) microorganisms to humans. The objective of the study was to assess the occurrence of extended-spectrum ß-lactamase (ESBL)- and carbapenemase-producing Enterobacterales (ESBL-E and CPE), respectively, in 118 fresh herbs and 101 bagged salads collected at retail level in Switzerland and to characterize the isolates' phenotypic and genotypic properties using culture-based methods and whole genome sequencing (WGS). Of the fresh herbs, 6/118 contained ESBL-E and 7/118 yielded CPE. Of the salads, 13/101 contained ESBL-E and 1/101 CPE. Antimicrobial susceptibility testing (AST) identified 9/29 isolates as multidrug-resistant (MDR). ESBL-E were Escherichia coli (n=6), Klebsiella pneumoniae (n=4) Enterobacter chuandaensis (n=1), and Kluyvera spp. (n=1) carrying ß-lactamase (bla) genes belonging to the cefotaximase-München (blaCTX-M)-groups, Proteus spp. (n=1) containing Hôpital- Universitaire-de-Genève-bla (blahugA), Raoultella ornithinolytica (n=1) carrying sulfhydryl reagent variable bla (blaSHV), and Serratia fonticola (n=7) carrying S. fonticula bla (blaFONA) genes. CPE were Enterobacter asburiae (n=1) E. cloacae (n=6) and E. vonholyi (n=1) carrying imipenemase bla (blaIMI) genes. Several K. pneumoniae sequence types (STs) were identified (ST967, ST628, ST219, and ST1823), which have been linked to human disease and nosocomial outbreaks. They carried blaCTX-M-15 on plasmids detected globally in environmental and clinical samples. E. coli (ST10, ST48, ST609, ST2040, ST6215 and ST3580) and enterotoxigenic E. coli (ETEC) ST2040 carrying blaCTX-M-15 were found. E. cloacae (ST820 and ST1516) with blaIMI-1 have been found previously in clinical settings and community outbreaks. The occurrence and consumption of fresh produce containing MDR ESBL-E and CPE pose substantial public health risks and raise significant food safety concerns.

A Taq-Man-based multiplex quantitative PCR for the simultaneous detection and quantification of Angiostrongylus vasorum, Crenosoma vulpis, and species of respiratory capillarids in canids.

In recent years, Angiostrongylus vasorum, Crenosoma vulpis, Eucoleus aerophilus (syn. Capillaria aerophila) and Eucoleus boehmi (syn. Capillaria boehmi), commonly referred to as canine lungworms, have gained a growing interest worldwide as the result of their geographical expansion. Each of these nematode species differs considerably in its biology and pathogenicity. Despite their impact on dogs' health, these parasites are often underdiagnosed owing to diagnostic challenges. Here, we describe the development and validation of a Taq-Man-based multiplex quantitative PCR (qPCR) for the simultaneous detection of the main species of canine lungworms in faeces of infected dogs. Using 10-fold serial dilutions of synthetic gene block fragments containing individual sequence targets of each lungworm species, the analytical sensitivity of the assay ascertained was 1.84 ng/µl for A. vasorum, 3.08 ng/µl for C. vulpis and 0.79 ng/µl for Eucoleus spp. The sensitivity of the assays and their ability to detect mixed species infections were compared with microscopy-based techniques (faecal floatation and Baermann technique) applied to faecal samples submitted for lungworm testing through an accredited diagnostic laboratory at the Institute of Parasitology, University of Zurich, Switzerland, and from community dogs as part of a research project on canine endoparasites in Cambodia. The multiplex qPCR displayed high diagnostic sensitivity (42/46, 91.3%; 95% Confidence Interval (CI): 79.1-97.1%) and a diagnostic specificity of 100% (45/45, 95% CI: 90.6-100%), and was able to detect 42.9% additional mixed lungworm species infections compared with microscopy-based methods. Kappa statistics showed substantial agreement between the qPCRs and microscopy for mixed infections (κ = 0.72, 95% CI: 0.4-1) and Eucoleus spp. (κ = 0.65, 95% CI: 0.45-0.85) and almost perfect agreement for C. vulpis (κ = 0.85, 95% CI: 0.63-1) and A. vasorum (κ = 0.92, 95% CI: 0.84-1). This multiplex qPCR enables timely, accurate, and sensitive diagnosis of canine lungworm species in faecal samples and can be used to monitor the geographical distribution and emergence of these parasitic species, globally.

Seek, and you will find: Cryptic diversity of the cardiopulmonary nematode Angiostrongylus vasorum in the Americas.

Angiostrongylus vasorum is a metastrongylid parasite infecting wild canids and domestic dogs. Its patchy distribution, high pathogenicity and taxonomical classification makes the evolutionary history of A. vasorum intriguing and important to study. First larval stages of A. vasorum were recovered from feces of two grey foxes, Urocyon cinereoargenteus, from Costa Rica. Sequencing and phylogenetic and haplotypic analyses of the ITS2, 18S and cytochrome oxidase subunit 1 (cox1) fragments were performed. Then p- and Nei´s genetic distance, nucleotide substitution rates and species delimitation analyses were conducted with cox1 data of the specimens collected herein and other Angiostrongylus spp. Cophylogenetic congruence and coevolutionary events of Angiostrongylus spp. and their hosts were evaluated using patristic and phenetic distances and maximum parsimony reconciliations. Specimens from Costa Rica clustered in a separate branch from European and Brazilian A. vasorum sequences in the phylogenetic and haplotype network analyses using the ITS2 and cox1 data. In addition, cox1 p-distance of the sequences derived from Costa Rica were up to 8.6 % different to the ones from Europe and Brazil, a finding mirrored in Nei´s genetic distance PCoA. Species delimitation analysis supported a separate group with the sequences from Costa Rica, suggesting that these worms may represent cryptic variants of A. vasorum, a new undescribed taxon or Angiocaulus raillieti, a synonym species of A. vasorum described in Brazil. Moreover, nucleotide substitution rates in A. vasorum were up to six times higher than in the congener Angiostrongylus cantonensis. This finding and the long time elapsed since the last common ancestor between both species may explain the larger diversity in A. vasorum. Finally, cophylogenetic congruence was observed between Angiostrongylus spp. and their hosts, with cospeciation events occurring at deeper taxonomic branching of host order. Altogether, our data suggest that the diversity of the genus Angiostrongylus is larger than expected, since additional species may be circulating in wild canids from the Americas.

Efficacy of Bravecto® Plus spot-on solution for cats (280 mg/ml fluralaner and 14 mg/ml moxidectin) in the prevention of feline Aelurostrongylus abstrusus infection evaluated in a multi-diagnostic approach.

BACKGROUND: Aelurostrongylus abstrusus is one of the most important respiratory nematodes of felines. Infections may lead to respiratory clinical signs with varying severity or even death, emphasizing the need for preventive treatment of cats with outdoor access to circumvent patent infections. METHODS: Therefore, the preventive efficacy of a spot-on formulation of 280 mg/ml fluralaner and 14 mg/ml moxidectin (Bravecto® Plus spot-on solution for cats, MSD) against A. abstrusus was evaluated in a negative controlled, randomized and partially blinded efficacy study with 28 purpose-bred cats in a non-terminal design. In three different treatment regimes, the minimum recommended dose of 40 mg fluralaner and 2.0 mg moxidectin/kg bodyweight (BW) was administered once at 12, 8 or 4 weeks (study group G1, G2 and G3, respectively) prior to experimental infection with 300 third-stage A. abstrusus larvae, while G4 served as placebo-treated control. RESULTS: From 30 to 46 days post infection (dpi; SD 114 to 130), faeces were sampled to monitor first-stage larvae (L1) excretion for efficacy determination. Secondary efficacy criteria, including respiratory parameters, serological antibody levels and computed tomography (CT) findings, were assessed once before enrolment (SD -7 to -1) and before infection (SD 75 to 83). After infection, CT evaluation was performed once at 47-50 dpi (SD 131 to 134), and respiratory parameters and antibody levels were regularly assessed twice or once a week, respectively (1 up to 78 dpi, SD 85 up to 162). All animals in the control group excreted L1 by 33-37 dpi and remained positive throughout the study period from 41 to 46 dpi (SD 125 to 130). In the treatment groups, only one animal each of G1 and G2 excreted L1 at two consecutive days, and four cats of G1, two of G2 and three of G3 were positive on single occasions. While the geometric mean (GM) of the maximum number of excreted L1 per 5 g of faeces was 7380.89 in the control group (G4), GMs were significantly lower in the treatment groups with 1.63 in G1, 1.37 in G2 and 0.79 in G3. Thus, based on GMs, the reduction in excreted L1 exceeded 99.9% in all three treatment groups. Based on CT severity scores, all lungs of the animals of the control group showed severe pulmonary changes post infection, whereas lungs of the cats of the treatment groups were either unaltered (4 animals), mildly (11 animals), or moderately altered (5 animals). Moreover, seroconversion was observed in all cats of the control group, but not in those of the treatment groups. CONCLUSIONS: The combination of diagnostic methods used in this non-terminal study yielded coherent and reliable results. A single administration of Bravecto® Plus spot-on solution for cats was well tolerated and effective in the prevention of aelurostrongylosis for at least 12 weeks.

Serological detection of circulating Angiostrongylus vasorum antigen- and parasite-specific antibodies in dogs from Poland.

Dogs infected with Angiostrongylus vasorum, a potentially lethal parasite living in the heart and pulmonary arteries, may present severe respiratory and neurological sings and coagulopathies. Its occurrence is increasingly reported from various European countries, but little is known about its presence in Poland. In this first large-scale survey, 3,345 sera from polish dogs attending veterinary clinics in different parts of Poland for various reasons were collected and tested by an ELISA for the detection of circulating antigen of A. vasorum and by a separate (n = 17, 95% Confidence Intervals, CI: 0.30­0.81 %) of the animals were positive in both ELISAs, while 0.78 % (n = 26, CI: 0.51­1.14 %) of the tested dogs were antigen- positive only and 1.29 % (n = 43, CI: 0.93­1.73 %) were positive for specific antibodies only. Regions with antigen- and antibody-positive animals were overlapping and distributed over the whole area of the country, with approximately one third of positives close to the Baltic Sea, and a limited number of cases close to the German border. These results confirm the occurrence of A. vasorum in dogs originating from different parts of Poland. A. vasorum serology presents significant advantages (diagnosis before patency, single serum sample instead of repeated faecal samples, rapidity and affordability particularly in case of large number of samples), and it can be considered a valid alternative for diagnosis in individuals and in epidemiological studies.

Canine angiostrongylosis in naturally infected dogs: clinical approach and monitoring of infection after treatment.

Canine angiostrongylosis is an increasingly reported disease in Europe which can be fatal if left untreated. The wide range of clinical presentation along with the absence of pathognomonic alterations can make the diagnosis challenging; thus any additional information that may provide clues to an early diagnosis may be of value, in order to ensure adequate anthelmintic treatment. Aim of the study was to assess a clinicopathological scoring system associated with natural Angiostrongylus vasorum infection diagnosed in canine patients during clinical practice, to clinically and paraclinically monitor infected dogs after treatment, and to monitor the presence of L1 larvae in faecal samples by Baermann's test. Of the total 210 enrolled animals A. vasorum infection was diagnosed in 7 dogs. These dogs were clinically and paraclinically investigated and monitored after specific treatment. Further 3 symptomatic dogs were retrospectively included in the monitoring. Results suggest that the computed scoring system can help to increase the clinical suspicion of infection particularly in asymptomatic dogs before the onset of potentially lethal lesions. Data of faecal monitoring suggested that treatment may control parasite burden but be unable to eradicate infection. Thus, a continued faecal monitoring after treatment is advisable for identification of still infected or reinfected dogs.