Fox Serum Proteomics Analysis Suggests Host-Specific Responses to Angiostrongylus vasorum Infection in Canids

Dogs infected with the cardiopulmonary nematode Angiostrongylus vasorum may suffer from respiratory distress and/or bleeding disorders. Descriptions of clinical signs in foxes are rare, despite high prevalence. To evaluate the impact of infection on coagulation and immune response, serum proteins from eight experimentally infected foxes before and after inoculation (day 0, 35, 84, 154) were subjected to differential proteomic analyses based on quantitative data and compared to available data from dogs. The number of proteins with differential abundance compared to the uninfected baseline increased with chronicity of infection. Bone marrow proteoglycan, chitinase 3-like protein 1 and pulmonary surfactant-associated protein B were among the most prominently increased proteins. The abundance of several proteins involved in coagulation was decreased. Enriched pathways obtained from both increased and decreased proteins included, among others, “platelet degranulation” and “haemostasis”, and indicated both activation and suppression of coagulation. Qualitative comparison to dog data suggests some parallel serum proteomic alterations. The comparison, however, also indicates that foxes have a more adequate immunopathological response to A. vasorum infection compared to dogs, facilitating persistent infections in foxes. Our findings imply that foxes may be more tolerant to A. vasorum infection, as compared to dogs, reflecting a longer evolutionary host–parasite adaptation in foxes, which constitute a key wildlife reservoir.

The Angiostrongylus vasorum Excretory/Secretory and Surface Proteome Contains Putative Modulators of the Host Coagulation

Angiostrongylus vasorum is a cardiopulmonary nematode of canids and is, among others, associated with bleeding disorders in dogs. The pathogenesis of such coagulopathies remains unclear. A deep proteomic characterization of sex specific A. vasorum excretory/secretory proteins (ESP) and of cuticular surface proteins was performed, and the effect of ESP on host coagulation and fibrinolysis was evaluated in vitro. Proteins were quantified by liquid chromatography coupled to mass spectrometry and functionally characterized through gene ontology and pathway enrichment analysis. In total, 1069 ESP (944 from female and 959 from male specimens) and 1195 surface proteins (705 and 1135, respectively) were identified. Among these were putative modulators of host coagulation, e.g., von Willebrand factor type D domain protein orthologues as well as several proteases, including serine type proteases, protease inhibitors and proteasome subunits. The effect of ESP on dog coagulation and fibrinolysis was evaluated on canine endothelial cells and by rotational thromboelastometry (ROTEM). After stimulation with ESP, tissue factor and serpin E1 transcript expression increased. ROTEM revealed minimal interaction of ESP with dog blood and ESP did not influence the onset of fibrinolysis, leading to the conclusion that Angiostrongylus vasorum ESP and surface proteins are not solely responsible for bleeding in dogs and that the interaction with the host’s vascular hemostasis is limited. It is likely that coagulopathies in A. vasorum infected dogs are the result of a multifactorial response of the host to this parasitic infection.

Highly Variable Clinical Pictures in Dogs Naturally Infected with Angiostrongylus vasorum

Canine angiostrongylosis by Angiostrongylus vasorum is increasingly reported in both enzootic and previously free areas. The complex pathogenesis of the disease makes the clinical workup challenging. Infected dogs show highly variable clinical pictures, characterized by subclinical to life-threatening general, cardio-respiratory, neurological and/or gastrointestinal signs. The present study reports the high variability of clinical pictures from 36 dogs across central and southern Italy that were naturally infected by A. vasorum. Of them, 23 (63.9%) presented at least one clinical sign, while 13 (36.1%) were subclinically infected and apparently healthy. Overall, 19 dogs (52.8%) showed cardiorespiratory signs, 14 (38.9%) had non-specific abnormalities, 2 (5.6%) presented coagulation disorders and 1 (2.8%) had a severe neurological condition. Importantly, four dogs presenting with clinical signs had neither cough nor dyspnea. These results underline that angiostrongylosis should be included in the differential diagnosis, even when dogs display only non-specific clinical signs. The proportion of apparently healthy dogs highlights the relevance of routine copromicroscopic and/or antigenic tests in enzootic areas to avoid the sudden onset of potentially life-threatening signs.

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.

Dirofilaria spp. and Angiostrongylus vasorum: Current Risk of Spreading in Central and Northern Europe

In the past few decades, the relevance of Dirofilaria immitis and Dirofilaria repens, causing cardiopulmonary and subcutaneous dirofilariosis in dogs and cats, and of Angiostrongylus vasorum, causing canine angiostrongylosis, has steadily increased in Central and Northern Europe. In this review, a summary of published articles and additional reports dealing with imported or autochthonous cases of these parasites is provided for Central (Austria, Czechia, Germany, Hungary, Luxemburg, Poland, Slovakia, Slovenia, and Switzerland) and Northern (Denmark, Finland, Iceland, Norway, and Sweden) Europe. Research efforts focusing on Dirofilaria spp. and A. vasorum have varied by country, and cross-border studies are few. The housing conditions of dogs, pet movements, the spread of competent vectors, and climate change are important factors in the spread of these nematodes. Dogs kept outside overnight are a major factor for the establishment of Dirofilaria spp. However, the establishment of invasive, diurnal, synanthropic, competent mosquito vectors such as Aedes albopictus may also influence the establishment of Dirofilaria spp. The drivers of the spread of A. vasorum remain not fully understood, but it seems to be influenced by habitats shared with wild canids, dog relocation, and possibly climatic changes; its pattern of spreading appears to be similar in different countries. Both Dirofilaria spp. and A. vasorum merit further monitoring and research focus in Europe.

Simultaneous Exposure to Angiostrongylus vasorum and Vector-Borne Pathogens in Dogs from Italy

Several drivers have recently fostered the expansion of Angiostrongylus vasorum throughout Europe, where Vector-Borne Pathogens (VBPs) are also spreading. However, the level of simultaneous risk of infection is still unknown in canine populations. This study evaluated the simultaneous exposure to A. vasorum and major canine VBPs in dogs of Italy. Sera of 294 dogs were subjected to two ELISAs, detecting A. vasorum circulating antigens and antibodies against the parasite, and to the following assays: (i) SNAP® 4DX (IDEXX Laboratories Inc.) detecting Dirofilaria immitis antigens, and antibodies vs. Borrelia burgdorferi, Anaplasma spp. and Ehrlichia spp. and (ii) IFAT for the detection of antibodies vs. Leishmania infantum, Babesia canis and Rickettsia conorii. Twenty-two (7.5%, CI: 4.8–11.1%) and six (2%, CI: 0.7–4.4%) dogs scored positive for circulating A. vasorum antibodies and antigens, respectively. Seventeen dogs (5.8%, CI: 3.4–9.1%) were positive for A. vasorum antibodies + at least one VBP, three (1%, CI: 0.2–3%) for A. vasorum antigen + at least one VBP, while one dog (0.3%, CI: 0.01–1.88%) was positive for A. vasorum antigen + A. vasorum antibodies + B. canis antibodies. These results show that dogs living in different regions of Italy are at risk of simultaneous infections with both A. vasorum and VBPs. Despite the same scenario being likely in other countries of Europe, the current knowledge is scant. Therefore, further studies are warranted to amplify current epizootiological information and to understand whether control programs should be improved.

ANGIOSTRONGILOSE CANINA: RELATO DE CASO

Introdução: Angiostrongylus vasorum é um nematoide parasito de canídeos domésticos e silvestres, cuja forma adulta pode ser encontrada no ventrículo direito, artérias pulmonares e ramificações. O ciclo do parasito é do tipo heteróxeno, seus hospedeiros definitivos são os canídeos e seus hospedeiros intermediários são moluscos terrestres e aquáticos que se infectam pela ingestão de larvas de primeiro estádio. Os canídeos podem ser infectados através da ingestão de hospedeiros paratênicos como anfíbios. Os sinais clínicos apresentados são: apatia, náusea, vômito, tosse, perda de peso, anorexia, ascite, menor tolerância a atividade física, dispneia intensa e dor abdominal. O diagnóstico clínico é feito através de testes laboratoriais juntamente com os sinais clínicos apresentados. No diagnóstico diferencial os principais achados são bronquite crônica e asma e pode ser utilizado os exames radiológicos, ecocardiográficos e eletrocardiográficos para o diagnóstico. Objetivo: relatar o caso clínico de um canino doméstico acometido por Angiostrongilose. Materiais e Métodos: revisão do prontuário, entrevista com tutor, registro fotográfico dos métodos diagnósticos, aos quais o paciente foi submetido e revisão da literatura. Resultados: as ocorrências de angiostrongilose na clínica médica são muito importantes para o conhecimento e no cotidiano do médico veterinário, além da contribuição para as estratégias adotadas para o diagnóstico e tratamento das doenças. A sintomatologia cardiorrespiratória pode ser justificada pela infiltração de células inflamatórias e broncopneumonia, resultantes da deposição de ovos e migração das larvas pela árvore brônquica do hospedeiro, nos achados radiográficos relacionados com o caso foi diagnosticado a broncopneumonia, e assim direcionado o diagnóstico. Conclusão: os sintomas de angiostrongilose canina pode ser confundida facilmente com outras enfermidades. Exames laboratoriais e de imagens são essenciais para o diagnóstico de angiostrongilose uma vez que sem eles os sinais clínicos apresentados são inconclusivos ao diagnóstico final.