pathogens Article Detection of Crenosoma spp., Angiostrongylus vasorum and Aelurostrongylus abstrusus in Gastropods in Eastern Austria 1, , 2, 1 1 Hans-Peter Fuehrer * y, Simone Morelli y , Julian Bleicher , Thomas Brauchart , Mirjam Edler 1, Nicole Eisschiel 1, Tatjana Hering 1, Sigrun Lercher 1, Karoline Mohab 1, Simon Reinelt 1, Theresa Stessl 1, Doris Fasching 1, Ricarda Nimphy 1, Anja Pelzl 1, Bita Shahi-Barogh 1, Licha Natalia Wortha 1, Karin Bakran-Lebl 1 , Michael Duda 3, Helmut Sattmann 3, Roland Schaper 4 , Donato Traversa 2 and Anja Joachim 1 1 Department of Pathobiology, Institute of Parasitology, University of Veterinary Medicine, 1210 Vienna, Austria; [email protected] (J.B.); [email protected] (T.B.); [email protected] (M.E.); [email protected] (N.E.); [email protected] (T.H.); [email protected] (S.L.); [email protected] (K.M.); [email protected] (S.R.); [email protected] (T.S.); [email protected] (D.F.); [email protected] (R.N.); [email protected] (A.P.); [email protected] (B.S.-B.); [email protected] (L.N.W.); [email protected] (K.B.-L.); [email protected] (A.J.) 2 Faculty of Veterinary Medicine, University of Teramo, 64100 Teramo, Italy; [email protected] (S.M.); [email protected] (D.T.) 3 Department of Invertebrate Zoology, Natural History Museum Vienna, 1010 Vienna, Austria; [email protected] (M.D.); [email protected] (H.S.) 4 Elanco Animal Health, 40789 Monheim, Germany; [email protected] * Correspondence: [email protected]; Tel.: +43-(1)-25077-2205 These two authors contributed equally to this work. y Received: 19 November 2020; Accepted: 11 December 2020; Published: 13 December 2020 Abstract: Canine and feline cardiorespiratory parasites are of utmost relevance in veterinary medicine. Key epizootiological information on major pet metastrongyloids, i.e., Angiostrongylus vasorum and Crenosoma vulpis infecting dogs, and Aelurostrongylus abstrusus and Troglostrongylus brevior infecting cats, is missing from Austria. This study investigated their occurrence in 1320 gastropods collected in the Austrian provinces of Styria, Burgenland, Lower Austria, and in metropolitan Vienna. Metastrongyloid larvae were microscopically detected in 25 samples, and sequence analysis confirmed the presence of metastrongyloids in nine samples, i.e., A. vasorum in one slug (Arion vulgaris) (0.07%), C. vulpis in five slugs (one Limax maximus and four A. vulgaris) (0.4%), A. abstrusus in two A. vulgaris (0.17%), and the hedgehog lungworm Crenosoma striatum was detected in one A. vulgaris. The present study confirms the enzooticity of major cardiorespiratory nematodes in Austria and that canine and feline populations are at risk of infection. Keywords: Angiostrongylus vasorum; Aelurostrongylus abstrusus; Crenosoma; Austria; PCR; Arion vulgaris 1. Introduction Cardiopulmonary metastrongyloid nematodes that affect dogs and cats are enzootic in Europe [1,2]. In the last years, these parasites have stimulated the interest of the veterinary scientific community for their emergence and clinical relevance [1,3,4]. Among them, Angiostrongylus vasorum and Crenosoma vulpis are the most relevant species infecting dogs [5,6], while Aelurostrongylus abstrusus and Troglostrongylus brevior are the most important species in cats [2,7]. Pathogens 2020, 9, 1046; doi:10.3390/pathogens9121046 www.mdpi.com/journal/pathogens PathogensPathogens 20202020, 9, ,x9 FOR, 1046 PEER REVIEW 2 of2 12 of 11 Angiostrongylus vasorum (the “French heartworm”) infects the pulmonary arteries of dogs throughoutAngiostrongylus Europe with vasorum a typical(the patchy “French geographic heartworm”)al distribution, infects constitute the pulmonaryd by endemic arteries foci of with dogs nearbythroughout low-prevalence Europe withareas a [8]. typical In the patchy last decade, geographical A. vasorum distribution, has spread constituted in various by endemicEuropean foci regions,with nearby in both low-prevalence enzootic areas areas [and8]. In areas the last previously decade, A. vasorumfree of hasinfection spread in[5,9,10]. various EuropeanCanine angiostrongylosisregions, in both may enzootic be fatal areas and and its areas clinical previously course is free unpredictable, of infection [as5,9 it,10 can]. Caninebe chronic, angiostrongylosis sub-clinic, acute,may or be hyperacute. fatal and its A clinical clinical course diagnosis is unpredictable, is almost impossible, as it can be as chronic, dogs infected sub-clinic, with acute, A. vasorum or hyperacute. can showA clinical a plethora diagnosis of nonspecific is almost as impossible,well as cardiopulmonary, as dogs infected neurological, with A. vasorum and gastrointestinalcan show a plethora clinical of signsnonspecific [3,11,12].as well as cardiopulmonary, neurological, and gastrointestinal clinical signs [3,11,12]. CrenosomaCrenosoma vulpis vulpis (“fox(“fox lungworm”) lungworm”) lives lives in in the bronchi,bronchi, bronchioles, bronchioles, and and trachea trachea of of dogs dogs and and other othercanids canids [13 ].[13]. This This nematode nematode occurs occurs at lower at lower prevalence prevalence rates rates than thanA. vasorumA. vasorumin Southern in Southern Europe Europe [8,14 ], [8,14],while while it is moreit is more prevalent prevalent in Central in Central and and Northern Northern Europe Europe [6,15 [6,15].]. Although Although dogs dogs may may display display severe severechronic chronic cough, cough, canine canine crenosomosis crenosomosis is rarely is rarely fatal fatal [1,6]. [1,6]. FelineFeline aelurostrongylosis aelurostrongylosis caused caused by byA. A.abstrusus abstrusus (“cat(“cat lungworm”) lungworm”) is distributed is distributed worldwide. worldwide. ThisThis nematode nematode infects infects bronchioles bronchioles and and alveolar alveolar ducts ducts of ofcats cats [16], [16 which], which may may be beeither either subclinically subclinically infectedinfected or orshow show respiratory respiratory or orgeneral general clinical clinical signs, signs, for for example, example, coug coughing,hing, sneezing, sneezing, wheezing, wheezing, lethargy,lethargy, depression, depression, and and occasionally, occasionally, death death [17–19]. [17–19 ]. DuringDuring the the past past decade, decade, T. breviorT. brevior hashas increasingly increasingly been been reported reported in domestic in domestic cats cats from from Europe, Europe, mainlymainly in incountries countries of of the the Mediterranean Mediterranean Basin Basin [20]. [20]. The natural hosthost ofofT. T. brevior brevioris is the the European European wild wildcat catFelis Felis silvestris, silvestris,but but it may it may also also infect infect domestic domestic cats, cats, causing causing severe severe infections infections especially especially in kittens in kittensand youngand young animals, animals, in which in which troglostrongylosis troglostrongylosis is often is often fatal [fatal20]. [20]. AllAll these these cardiopulmonary cardiopulmonary nematodes nematodes have have an anindirect indirect lifecycle, lifecycle, with with gastropods gastropods acting acting as as obligateobligate intermediate intermediate hosts hosts [1,20]. [1,20]. Surveys onon the the presence presence of metastrongyloidof metastrongyloid larvae larvae in field-collected in field- collectedintermediate intermediate hosts is ahosts useful is approacha useful approach for evaluating for evaluating the occurrence the occurrence of cardiopulmonary of cardiopulmonary nematodes in a nematodesgiven area in [ 21a given–23] and area predicting [21–23] and/assessing predicting/assessing the risk of infection the risk for canineof infection and feline for canine populations and feline [22,24 ]. populationsIn recent years, [22,24]. surveys In recent on larvalyears, nematodes surveys on harbored larval nematodes by wild-caught harbored mollusks by wild-caught have increasingly mollusks been haveperformed, increasingly mostly been in Europeperformed, [22,23 ,mostly25] and Southin Europe America [22,23,25] [21,26]. and Nevertheless, South America knowledge [21,26]. of the Nevertheless,presence and knowledge distribution of ofthe cardiopulmonary presence and dist nematodesribution of in bothcardiopulmon definitiveary and nematodes intermediate in hostsboth is definitivestill incipient and intermediate and requires hosts scientific is still refinements incipient in and certain requires European scientific countries, refinements such as in Austria certain [22 ]. EuropeanGiven the countries, great veterinary such relevanceas Austria of pet [22]. cardiopulmonary Given the nematodesgreat veterinary and the apparentrelevance geographical of pet cardiopulmonaryexpansion of extraintestinal nematodes and parasitoses the apparent of dogs geogra and catsphical due expansion to various of factors extraintestinal [8,16,24,27 parasitoses], the present of studydogs and investigated cats due to the various occurrence factors of [8,16,24,2 major canine7], the and present feline study metastrongyloids investigated the in snailsoccurrence and slugsof majorin Austria. canine and feline metastrongyloids in snails and slugs in Austria. 2. Results2. Results Overall,Overall, 1320 1320 gastropods gastropods belonging belonging to 12 to different 12 different species species were were collected collected (Figure (Figure 1 and1 and Table Table 1).1 ). Arion vulgaris