The current situation of Angiostrongylus vasorum in Romania: a national questionnaire-based survey.

BACKGROUND: Angiostrongylus vasorum (Nematoda, Metastrongyloidea) is a vascular nematode that resides in the pulmonary arteries and the right side of the heart of a wide variety of carnivores, with an indirect life cycle using coprophagic gastropods as intermediate hosts. For domestic dogs, the infection with A. vasorum can be asymptomatic, but more frequently, it is associated with a wide range of clinical manifestations like cardio-respiratory signs, bleedings, neurological signs, and ocular problems which can lead to death when not treated accordingly. Angiostrongylosis was confirmed for the first time in Romania in red foxes (Vulpes vulpes) in 2017 and two years later a seroepidemiologic study was conducted among domestic dogs. However, to this date, no clinical canine angiostrongylosis cases were published in Romania. The aim of the present paper was to evaluate the knowledge about canine angiostrongylosis among veterinarians in Romania and to update the distribution of this disease using a national wide anonymous questionnaire. RESULTS: Overall, 147 unique responses were submitted, from 31 out of 42 counties. Twelve veterinarians (8%) from 8 counties (26%) acknowledged diagnosing a case of angiostrongylosis including 5 from the Bucharest and 1 from each of the remaining seven counties. All affected dogs had respiratory distress, 75% suffered cardiopathy, 16% coagulopathies and 8% neurological signs. Case diagnosis was based mostly on larval detection by coprology (67%) and serological antigen detection test (42%). CONCLUSIONS: Romanian veterinarians are aware of canine angiostrongylosis and a significant number have clinical experience with the disease. Epidemiological studies are now needed to assess its distribution in the country, and further efforts are required to improve understanding of the disease, its diagnostic and treatment methods among veterinarians.

Autochthonous Angiostrongylus cantonensis, Angiostrongylus vasorum and Aelurostrongylus abstrusus infections in native terrestrial gastropods from the Macaronesian Archipelago of Spain.

The presence of zoonotic relevant Angiostrongylus cantonensis infections has recently been reported in rat final hosts and gastropod intermediate hosts in Tenerife, Spain. However, data on A. cantonensis, Angiostrongylus vasorum and Aelurostrongylus abstrusus prevalences in endemic gastropods for other islands of the Macaronesian Archipelago are still missing. In order to fill this gap, we conducted an epidemiological study on terrestrial native slug (Plutonia lamarckii) and snail (Cornu aspersum, Theba pisana, Rumina decollata) species in 27 selected locations of Tenerife, Gran Canaria, El Hierro, Lanzarote, La Palma and Fuerteventura. Overall, 131 terrestrial gastropods were collected in winter/spring season 2018/2019 and examined for the presence of metastrongyloid lungworm larvae via artificial digestion. The current data revealed a total prevalence of 4.6% for A. vasorum, 3.8% for A. abstrusus and 0.8% for A. cantonensis. In Tenerife, three lungworm species were detected, thereby re-confirming A. cantonensis endemicity for this island. Prevalences of snails (C. aspersum) originating from El Hierro were 5% for A. abstrusus and 15% for A. vasorum, respectively, with larval burdens up to 290 larvae per specimen. This epidemiological study indicates the presence of human, canine and feline lungworm species in Macaronesia, Spain. The current data-particularly those on anthropozoonotic A. cantonensis-call for a regular large-scale monitoring on intermediate hosts, paratenic hosts and definitive hosts to prevent further spread of lungworm-related diseases in humans and animals.

Infectivity of gastropod-shed third-stage larvae of Angiostrongylus vasorum and Crenosoma vulpis to dogs.

BACKGROUND: Metastrongyloid parasites Angiostrongylus vasorum and Crenosoma vulpis infect wild and domestic canids and are important pathogens in dogs. Recent studies indicate that gastropod intermediate hosts infected with various metastrongyloids spontaneously shed infective third-stage larvae (L3) into the environment via feces and mucus under laboratory conditions. Shed L3 retain motility up to 120 days, but whether they retain infectivity was unknown. METHODS: To assess the infectivity of shed L3, the heart/lungs of six red foxes (Vulpes vulpes) were obtained from trappers in Newfoundland, Canada. Lungs were examined for first-stage larvae (L1) by the Baermann technique. A high number of viable A. vasorum L1 and a low number of C. vulpis L1 were recovered from one fox; these were used to infect naïve laboratory-raised Limax maximus. L3 recovered from slugs by artificial digestion were fed to two naïve purpose-bred research beagles (100 L3/dog). L1 shed by these two dogs was used to infect 546 L. maximus (2000-10,000 L1/slug). L3 shedding was induced by anesthetizing slugs in soda water and transferring them into warm (45 °C) tap water for at least 8 h. Shed L3 recovered from slugs were aliquoted on romaine lettuce in six-well tissue culture plates (80-500 L3/well) and stored at 16 °C/75% relative humidity. Four naïve research beagles were then exposed to 100 L3/dog from larvae stored for 0, 2, 4, or 8 weeks, respectively, after shedding. RESULTS: All four dogs began shedding C. vulpis L1 by 26-36 days post-infection (PI). All four dogs began shedding A. vasorum L1 by 50 days PI. CONCLUSIONS: L3 infectivity for the definitive host was retained in both metastrongyloids, indicating the potential for natural infection in dogs through exposure from environmental contamination. As an additional exposure route, eating or licking plant or other material(s) contaminated with metastrongyloid L3 could dramatically increase the number of dogs at risk of infection from these parasites.

Morphological and molecular diagnosis of diseases of free-ranging crab-eating foxes (Cerdocyon thous) / [Diagnóstico morfológico e molecular de doenças de cachorro-do-mato (Cerdocyon thous) de vida livre]

The aim of this study was to investigate the occurrence of diseases in free-ranging wild canids that were roadkill on highways in the State of Espírito Santo, Brazil. PCR tests were performed for the detection of Brucella sp., Babesia sp., Rangelia sp., and Hepatozoon sp. in the spleen. Morphological evaluation and identification of parasites was performed in the liver and lung. Twenty specimens of C. thous were necropsied at the Animal Pathology Sector of the Veterinary Hospital of the Universidade Federal do Espírito Santo. Tissue samples were processed for histopathological examination and polymerase chain reaction (PCR) analysis. There was no PCR amplification of genomic DNA sequences of Brucella sp., Babesia sp., Rangelia sp., and Hepatozoon sp. using DNA extracted from the spleen as template. Histologically, lesions associated with parasitism by Platynosomum sp. and Angiostrongylus sp. were observed in the liver and lung, respectively. This is the first report of Platynosomum sp. and Angiostrongylus sp. parasitism in C. thous in the state of Espírito Santo, Brazil. Therefore, this study demonstrated parasitism of crab-eating foxes by Platynosomum sp. and Angiostrongylus sp. Importantly, no evidence of infection with Brucella sp., Babesia sp., Rangelia sp., and Hepatozoon sp. was obtained by PCR analysis.(AU)

O objetivo deste estudo foi investigar a ocorrência de doenças em canídeos silvestres de vida livre que foram atropelados em rodovias no estado do Espírito Santo, Brasil. Testes de PCR foram realizados para a detecção de Brucella sp., Babesia sp., Rangelia sp. e Hepatozoon sp. no baço. A avaliação morfológica e a identificação de parasitas foram realizadaa no fígado e no pulmão. Vinte espécimes de C. thous foram necropsiados no Setor de Patologia Animal do Hospital Veterinário da Universidade Federal do Espírito Santo. Amostras de tecido foram processadas para exame histopatológico e análise de reação em cadeia da polimerase (PCR). Não houve amplificação por PCR das sequências de DNA genômico de Brucella sp., Babesia sp., Rangelia sp. e Hepatozoon sp. usando-se DNA extraído do baço como modelo. Histologicamente, lesões associadas ao parasitismo por Platynosomum sp. e Angiostrongylus sp. foram observadas no fígado e no pulmão, respectivamente. Este é o primeiro relato de Platynosomum sp. e Angiostrongylus sp. parasitismo em C. thous no estado do Espírito Santo, Brasil. Portanto, este estudo demonstrou parasitismo de cachorro-do-mato por Platynosomum sp. e Angiostrongylus sp. É importante detacar que não há evidências de infecção por Brucella sp., Babesia sp., Rangelia sp. e Hepatozoon sp. por análise de PCR.(AU)

Investigation of the occurrence of Angiostrongylus vasorum in coyotes in southern Ontario, Canada.

In North America, the only endemic focus for Angiostrongylus vasorum (French heartworm) was historically thought to occur in the southeastern part of the island of Newfoundland. However, reports of A. vasorum infection in wild canids in West Virginia, USA, and Nova Scotia, Canada, suggest the introduction of the parasite to mainland North America. We screened for A. vasorum in coyotes from across southern Ontario. Additionally, we evaluated the performance of ELISAs for detection of circulating A. vasorum antigen (Ag-ELISA) and antibodies against A. vasorum (Ab-ELISA) designed for use in sera or blood of foxes for use with coyotes in this region. Autopsies were performed on 397 coyotes, and lung tissue extract prepared from each carcass was tested via both ELISAs. The sensitivity and specificity for both tests were estimated in the absence of a gold standard using a 2-test single population Bayesian model; sensitivity and specificity priors were based on the performance of the assays in foxes in Switzerland. Eight coyotes tested positive for A. vasorum antigen; no animal was antibody positive. The estimated sensitivity and specificity of the Ag-ELISA were 90.8% (95% credible interval [CrI]: 83.8-95.6%) and 95.5% (95% CrI: 93.4-97.2%), respectively. For the Ab-ELISA, the estimated sensitivity and specificity were 41.9% (95% CrI: 32.1-51.9%) and 98.0% (95% CrI: 96.3-99.0%), respectively. Based on these findings and negative postmortem data for the same animals, there is insufficient evidence to suggest the presence of A. vasorum in southern Ontario coyotes.

Gastropods as intermediate hosts of Angiostrongylus spp. in the Americas: bioecological characteristics and geographical distribution.

BACKGROUND: Intermediate hosts are key organisms in maintaining parasite life cycles, because they can act as amplifiers in the transmission from natural reservoirs to humans. One of the most important groups of intermediate hosts for zoonotic nematode infections are gastropods,slugs and snails. These are essential organisms in the larval development of Angiostrongylus species. OBJECTIVES: The objective of this paper is to review reports of Angiostrongylus spp. in naturally infected gastropods from the Americas, taking into account the diagnostic methods used in their identification, to be able to provide more accurate list of their intermediate hosts. We also discuss the factors that aid the dispersion of Angiostrongylus spp. in the Americas. METHODS: This study reviews scientific publications and book sections on Angiostrongylus spp. in the Americas, including original works assessing larvae of Angiostrongylus in intermediate hosts. The eligible reports were classified accordingly to their geographical location, year of first record, and the larvae identification methodologies used. Digital repositories were used for the search. The bioecological characteristics of the main intermediate hosts are summarised. FINDINGS: A total of 29 gastropod species that are naturally infected with Angiostrongylus spp. have been reported as intermediate hosts, 16 of which are land snails, two are freshwater snails, and 11 land slugs. MAIN CONCLUSIONS: This study highlights the importance of integrative studies, analysing both the etiological agent and its transmission dynamic in the environment, the biological and ecological characteristics of the hosts, and the impact on host populations. It is necessary to increase interdisciplinary studies to determine the potential epidemiological health risk of angiostrongyliasis in the Americas, and thus be able to establish prevention, monitoring and contingency strategies in the region.

A two-step morphology-PCR strategy for the identification of nematode larvae recovered from muscles after artificial digestion at meat inspection.

To ensure that meat from livestock and game is safe for human consumption, European legislation lays down rules for mandatory testing. Helminth larvae are a category of zoonotic foodborne pathogens that can contaminate meat. Among helminths, the only zoonotic nematode regulated in Europe regarding meat inspection is Trichinella spp.. It is precisely during Trichinella testing that other potentially zoonotic larvae can be found. Due to current lack of tools, their identification is often very complicated. Nematode larvae other than Trichinella, recovered from artificial digestions of pig and wild boar muscles from France and Germany, were subjected to a newly developed two-step identification scheme, which includes both morphological examination and molecular assays. The first step is a general orientation towards a broad taxonomic group; the second step consists of targeted identification based on the results of first step. Different parasites were identified, some of which were not zoonotic such as Metastrongylus spp. and Angiostrongylus vasorum, but others are known to be zoonotic such as Toxocara cati, Ascaris suum, and Uncinaria stenocephala. The strategy is efficient for the identification of nematode larvae recovered from muscles but could also be applied for larvae from other sources.

Intermediate gastropod hosts of major feline cardiopulmonary nematodes in an area of wildcat and domestic cat sympatry in Greece.

BACKGROUND: The metastrongyloid nematodes Aelurostrongylus abstrusus, Troglostrongylus brevior and Angiostrongylus chabaudi are cardiopulmonary parasites affecting domestic cats (Felis catus) and wildcats (Felis silvestris). Although knowledge on these nematodes has been improved in the past years, gaps in our knowledge of their distribution and role of gastropods as intermediate hosts in Europe still exist. This study reports on the presence of these nematodes and their intermediate hosts in an area in Greece where domestic cats and wildcats occur in sympatry. METHODS: Terrestrial gastropods were collected in the field and identified morphologically and by mitochondrial DNA-sequence analysis. Metastrongyloid larvae were detected by artificial digestion, morphologically identified to the species and stage level and their identity was molecularly confirmed. RESULTS: Aelurostrongylus abstrusus was found in the snails Massylaea vermiculata and Helix lucorum, T. brevior in the slug Tandonia sp., and A. chabaudi in the slug Limax sp. and the snails H. lucorum and M. vermiculata. CONCLUSIONS: To the best of our knowledge this study provides the first reports of (i) terrestrial gastropods being naturally infected with A. chabaudi, (ii) T. brevior naturally infecting terrestrial gastropods in Europe, and (iii) A. abstrusus naturally infecting terrestrial gastropods in Greece. Furthermore, the present study describes for the first time developmental stages of A. chabaudi and T. brevior in naturally infected gastropods. The biological characteristics of various intermediate gastropod hosts that could influence the distribution and expansion of feline cardiopulmonary nematodes are discussed, along with epizootiological implications and perspectives.

Abdominal angiostrongyliasis, report of two cases and analysis of published reports from Colombia. / Abdominal angiostrongyliasis, report of two cases and analysis of published reports from Colombia

Abdominal angiostrongyliasis is a parasitic zoonosis, endemic in the American continent. Its etiological agent is Angiostrongylus costaricensis, a nematode whose definitive hosts are rats and other rodents and the intermediate hosts, slugs. Mammals acquire the infection by consuming vegetables contaminated with L3 larvae. The disease shows a heterogeneous clinical spectrum and given its low incidence its diagnosis is a great challenge. In Colombia, the first case was reported in 1979 and until 1998, only five additional cases have been reported. However, in the last two decades, no new cases were reported. Here we discuss two cases of children from Huila and Caquetá departments who developed the disease. Both cases required long in-patient care and multiple surgical interventions. The diagnosis was achieved by histopathological observation of parasitic elements inside the mesenteric arteries. One of the children died while the other fully recovered. We discuss the epidemiology, pathogenic cycle, clinical presentation, diagnosis, and prevention strategies of this disease paying particular attention to our patients' features and the Colombian context.

La angiostrongiloidiasis abdominal es una zoonosis parasitaria endémica en el continente americano. Su agente etiológico es el nematodo Angiostrongylus costaricensis, cuyos huéspedes definitivos son los roedores y, los intermediarios, los caracoles y las babosas, por lo que se adquiere al consumir vegetales contaminados con larvas en estadio 3. La presentación clínica es muy variada y, dada su rareza, su diagnóstico es un desafío. En Colombia el primer caso se reportó en 1979 y, desde entonces hasta 1998, se han informado cinco casos más, aunque en las últimas dos décadas no se había reportado ningún caso. Se describen aquí dos casos de angiostrongiloidiasis en niños provenientes de los departamentos de Huila y Caquetá que requirieron una larga hospitalización y múltiples intervenciones quirúrgicas. El diagnóstico se logró al observar en los especímenes quirúrgicos larvas and huevos cuya morfología sugería una infección por nematodos; uno de los pacientes murió y el otro se recuperó satisfactoriamente. Se discuten la epidemiología, la patogenia, la presentación clínica, el diagnóstico y las estrategias de prevención de esta parasitosis, con énfasis en las características particulares de los casos descritos y en el contexto colombiano.

Development of a multiplex qPCR-based approach for the diagnosis of Dirofilaria immitis, D. repens and Acanthocheilonema reconditum.

BACKGROUND: Dirofilaria immitis, D. repens and Acanthocheilonema reconditum are the main causative agents of zoonotic canine filariosis. METHODS: We developed a combined multiplex approach for filaria and Wolbachia detection using the 28S-based pan-filarial and 16S-based pan-Wolbachia qPCRs, respectively, involving a fast typing method of positive samples using triplex qPCR targeting A. reconditum, D. immitis and D. repens, and a duplex qPCR targeting Wolbachia of D. immitis and D. repens. The approach was complemented by a duplex qPCR for the differential diagnosis of heartworms (D. immitis and Angiostrongylus vasorum) and pan-filarial cox1 and pan-Wolbachia ftsZ PCRs to identify other filarial parasites and their Wolbachia, respectively. A total of 168 canine blood and sera samples were used to validate the approach. Spearman's correlation was used to assess the association between filarial species and the strain of Wolbachia. Positive samples for both the heartworm antigen-test after heating sera and at least one DNA-positive for D. immitis and its Wolbachia were considered true positive for heartworm infection. Indeed, the presence of D. repens DNA or that of its Wolbachia as well as A. reconditum DNA indicates true positive infections. RESULTS: The detection limit for Wolbachia and filariae qPCRs ranged from 5 × 10-1 to 1.5 × 10-4 mf/ml of blood. When tested on clinical samples, 29.2% (49/168) tested positive for filariae or Wolbachia DNA. Filarial species and Wolbachia genotypes were identified by the combined multiplex approach from all positive samples. Each species of Dirofilaria was significantly associated with a specific genotype of Wolbachia. Compared to the true positives, the approach showed excellent agreement (k = 0.98-1). Unlike D. immitis DNA, no A. vasorum DNA was detected by the duplex qPCR. The immunochromatographic test for heartworm antigen showed a substantial (k = 0.6) and a weak (k = 0.15) agreements before and after thermal pre-treatment of sera, respectively. CONCLUSIONS: The proposed approach is a reliable tool for the exploration and diagnosis of occult and non-occult canine filariosis. The current diagnosis of heartworm disease based on antigen detection should always be confirmed by qPCR essays. Sera heat pre-treatment is not effective and strongly discouraged.

The 'French heartworm' in Greece: A countrywide serological survey of Angiostrongylus vasorum infection by combined detection of circulating antigens and specific antibodies.

Angiostrongylus vasorum is a nematode of the superfamily Metastongyloidea and was first described in the south of France in the nineteenth century where its biology was extensively studied in the 1960ies, leading to the common use of the byname "the French heartworm". The life cycle of the parasite is indirect including slugs and snails as intermediate hosts, while the adults live in the right heart and pulmonary arteries. Since then, many studies have been conducted in different European countries. However, there is scant knowledge regarding the prevalence and distribution of this parasite in Greece. Therefore, the objective of the present study was to investigate the current seroprevalence of A. vasorum in dogs in Greece. Overall 1000 apparently healthy and randomly selected dogs, presented in veterinary clinics, were involved at national level (n = 66 locations). Individual serum samples were obtained and tested using two in-house ELISAs by combining the detection of specific antibodies against the parasite and of circulating antigens of A. vasorum. Overall, 3.0% (n = 30, CI 95% 2.1 to 4.3%) of the sampled dogs were seropositive to at least one of the two ELISAs: 0.4% (n = 4, CI 95% 0.2 to 1.0%) were positive by both ELISAs, while 1.0% (n = 10, CI 95% 0.5 to 1.8%) were seropositive only for specific antibodies and 1.6% (n = 16, CI 95% 1.0 to 2.6%) were antigen-positive only. This is the first multicentric seroepidemiological study about A. vasorum in dogs from Greece. We showed that this parasite circulates in the local canine population and further studies should be carried out to elucidate its significance in the country.

Why Use the Mini-FLOTAC to Detect Metastrongyloid Larvae in Dogs and Cats?

PURPOSE: Canine and feline cardiopulmonary disease is an emergent condition in several European countries, particularly in non-endemic regions, although it is often underestimated because of the limitations of the common diagnostic techniques. The aim of this study was to evaluate the performance of Mini-FLOTAC to detect cardiopulmonary larvae from dogs and cats compared to reference techniques such as Baermann and FLOTAC. METHODS: A total of 20 fresh faecal samples were collected from ten dogs and ten cats naturally infected with Angiostrongylus vasorum, and Aelurostrongylus abstrusus and Troglostrongylus spp., respectively. Three replicates were performed for each technique. FS3 (zinc sulfate, specific gravity = 1.200) was used as flotation solution for Mini-FLOTAC and FLOTAC. The results were expressed as the arithmetic mean of LPG (larvae per gram of faeces). RESULTS: The mean LPG calculated by means of Mini-FLOTAC was significantly higher (P < 0.05) than that obtained through the Baermann technique for A. vasorum, A. abstrusus and Troglostrongylus spp., whereas it was significantly higher (P < 0.05) than that obtained through the FLOTAC technique only for A. vasorum and A. abstrusus. CONCLUSIONS: Mini-FLOTAC can be considered a valid alternative for the detection of metastrongyloid larvae of dogs and cats, overcoming the limitation of time required by the Baermann test.

Gastropods as intermediate hosts of Angiostrongylus spp. in the Americas: bioecological characteristics and geographical distribution

BACKGROUND Intermediate hosts are key organisms in maintaining parasite life cycles, because they can act as amplifiers in the transmission from natural reservoirs to humans. One of the most important groups of intermediate hosts for zoonotic nematode infections are gastropods,slugs and snails. These are essential organisms in the larval development of Angiostrongylus species. OBJECTIVES The objective of this paper is to review reports of Angiostrongylus spp. in naturally infected gastropods from the Americas, taking into account the diagnostic methods used in their identification, to be able to provide more accurate list of their intermediate hosts. We also discuss the factors that aid the dispersion of Angiostrongylus spp. in the Americas. METHODS This study reviews scientific publications and book sections on Angiostrongylus spp. in the Americas, including original works assessing larvae of Angiostrongylus in intermediate hosts. The eligible reports were classified accordingly to their geographical location, year of first record, and the larvae identification methodologies used. Digital repositories were used for the search. The bioecological characteristics of the main intermediate hosts are summarised. FINDINGS A total of 29 gastropod species that are naturally infected with Angiostrongylus spp. have been reported as intermediate hosts, 16 of which are land snails, two are freshwater snails, and 11 land slugs. MAIN CONCLUSIONS This study highlights the importance of integrative studies, analysing both the etiological agent and its transmission dynamic in the environment, the biological and ecological characteristics of the hosts, and the impact on host populations. It is necessary to increase interdisciplinary studies to determine the potential epidemiological health risk of angiostrongyliasis in the Americas, and thus be able to establish prevention, monitoring and contingency strategies in the region.

First case of a natural infection in a domestic cat (Felis catus) with the canid heart worm Angiostrongylus vasorum.

Cardiopulmonary nematodes in cats include different parasite species affecting feline lungs and the heart, with the metastrongyloid Aelurostrongylus abstrusus being the most frequent feline lungworm worldwide. The present case report describes an 11-month-old male neutered European short hair cat which presented with generalised subcutaneous oedema and pleural and peritoneal effusions. According to clinical examination, abdominal imaging and laboratory analyses, a tentative diagnosis of severe glomerulopathy with massive proteinuria was made. Due to worsening of the clinical signs despite therapeutic interventions and a poor prognosis, the cat was euthanised. Necropsy and histological examinations revealed severe bilateral collagenofibrotic glomerulopathy, generalised oedema and a focal verminous pneumonia with thrombosis in arterial lung vessels containing nematode cross sections. A serum sample was tested for the presence of antibodies against the cat lungworm A. abstrusus, resulting negative. Genetic analyses confirmed the presence of nematode DNA; after exclusion of common lung and heart parasites occurring in cats, DNA of the canid heart worm nematode Angiostrongylus vasorum was identified. This is the first description of a naturally occurring infection with A. vasorum in a cat. Previous experimental studies demonstrated the development of adult male and female A. vasorum worms containing eggs in cats, but no larval excretion in the faeces. Although cats did not become patent, A. vasorum infections were clinically relevant. As A. abstrusus and A. vasorum are both gastropod transmitted nematodes, they may share the same intermediate hosts within overlapping areas. In addition, especially chronic A. abstrusus infected cats become non-patent and do not excrete L1. Considering that patent A. vasorum infections are widespread in the dog and fox population in Switzerland (and several other countries) but are apparently not patent in cats, we cannot exclude that infections with A. vasorum may occur more frequently than expected.

Irreversible Ocular Lesions in a Dog With Angiostrongylus Vasorum Infection.

This report describes a 10-month-old dog with a sudden loss of vision and severe dyspnoea. The ocular examination revealed bilateral panuveitis, lens subluxation, secondary glaucoma, and retinal detachment. In addition, the ocular ultrasound showed in the vitreous body of the right eye, a small doubled-lined foreign body compatible with an intraocular parasite. Radiographs of the thorax revealed an increased opacity with mixed lung pattern (alveolar and bronchial) and thoracic ultrasonography showed several subpleural nodules. The presence of Angiostrongylus vasorum first stage larvae was confirmed with 324 larvae per gram of feces and an antigen test for the parasite (AngioDetect, IDEXX) also yielded a positive result. The severe and irreversible ocular lesions described in this case enhanced the complexity of the clinical picture of canine angiostrongylosis. Infection with the parasite should be included in the list of differential diagnoses for ocular uveitis to avoid potentially serious complications related to a missed or delayed diagnosis.

Are golden jackals (Canis aureus) definitive hosts for Angiostrongylus vasorum?

Angiostrongylosis caused by Angiostrongylus vasorum is an emerging disease in Europe. Recent reports have shown that, besides the red fox (Vulpes vulpes) recognized as the main definitive host and reservoir for the parasite, the role of the definitive host can be taken by a range of mammals. We considered that, due to the rapid large-scale expansion of its populations in Europe, the golden jackal (Canis aureus) could assume an important role in the epidemiology of the disease. The aim of the investigations was to explore the role of the golden jackal as the definitive host for A. vasorum. Sixty-three golden jackals, legally hunted in lowlands around the Danube River in Serbia recognized as the core area of the species' distribution in Europe, were subjected to patho-morphological and parasitological examination. The adult forms of A. vasorum were detected in the pulmonary arteries in six golden jackals with gross lesions manifested in the lungs. The finding of first stage larvae (L1) of A. vasorum in microscopic smears of the lung tissue altered by infection, which was consistent with the presence of adult parasites and manifestation of gross lesions in the lungs, confirms the successful reproduction of the parasite in the golden jackal. Migration of L1 from the blood vessels to the airways was confirmed by histopathology and, subsequently, their shedding was demonstrated by the Baermann method. The results support the hypothesis that the golden jackal acts as a suitable definitive host for A. vasorum. As a definitive host with a large-scale expansion of its populations in Europe, the golden jackal may be an important part of the parasite's host repertoire by spreading the parasite into previously non-endemic areas and by being an additional definitive host in endemic areas of vulpine angiostrongylosis.

The first seroepidemiological survey for Angiostrongylus vasorum in domestic dogs from Romania.

BACKGROUND: Angiostrongylus vasorum is a metastrongyloid nematode localized in the right heart and the pulmonary arteries of domestic dogs. The number of reports in Europe has recently increased, presumably as a consequence of a growing awareness among clinicians, animal owners and researchers, but also due to a growing incidence and territorial spread. So far, no studies have been conducted to assess the prevalence and distribution of A. vasorum in domestic dogs in Romania, and the awareness among veterinarians is limited or absent. The aim of the present study was to evaluate the countrywide seroprevalence of circulating antigens of A. vasorum and specific antibodies against A. vasorum in domestic dogs from Romania. METHODS: Between November 2016 and July 2017, blood was sampled from a total of 1545 domestic dogs from 23 counties of Romania. Details about their gender, age, breed, housing, use and origin were collected. All serum samples were tested for the presence of A. vasorum circulating antigens (AG) using monoclonal and polyclonal antibodies in a sandwich ELISA. Additionally, a sandwich ELISA using A. vasorum adult somatic antigen purified by monoclonal antibodies was used for specific antibody (AB) detection. RESULTS: A total of 33 dogs (2.14%, 95% CI: 1.82-3.56%) were seropositive for A. vasorum antigen or antibodies against the parasite. Three dogs were positive for antigen only (0.19%, 95% CI: 0.07-0.57%) and 30 dogs (1.94%, 95% CI: 1.36-2.76%) were positive for antibodies only. No dog was positive for both tests. The overall prevalence (AB or AG) and the AB prevalence were significantly higher in pure breed dogs compared to mixed breeds and mongrel dogs (P < 0.05) and in shepherd dogs compared to other groups (P < 0.05). There was no significant difference between males and females, between urban and rural dogs, between dogs with unrestricted access and with restricted access to the environment, and between dogs living outdoors and indoors. CONCLUSIONS: Our data suggest that the disease is present in Romania in dogs, as it was previously demonstrated in foxes. However, so far, no clinical case has been reported in the country and this may be related to a low awareness among vets.

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.

Molecular detection of Angiostrongylus vasorum in gastropods in Surrey, UK.

Surrey, a county in southern England, is a hot spot for angiostrongylosis in domestic dogs but there have been no investigations into the intermediate hosts of Angiostrongylus vasorum in this area. This study aimed to determine the prevalence of A. vasorum in gastropods in Guildford, the most populous town in Surrey, and to ascertain which gastropod species can act as intermediate hosts for A. vasorum. Gastropods (n = 97) were sampled in six locations, representing urban, suburban and rural environments, and identified to species based on morphological features. A PCR assay was used to detect A. vasorum DNA in gastropod tissue and the species of infected specimens was confirmed by sequencing of mitochondrial genes. 4.1% (4/97) of sampled gastropods and 9.1% (4/44) of sampled slugs were A. vasorum positive. Infected gastropod species were Arion rufus (n = 3) and Deroceras invadens (n = 1), the first description of the latter species as a potential intermediate host for A. vasorum. Two infected slugs were sampled in urban environments and two in suburban environments. The results demonstrate that there is a risk of transmission of A. vasorum to domestic dogs from the gastropod population in urban and suburban areas of Guildford.