Two cases of Angiostrongylus vasorum, a cardiopulmonary nematode, in a wild black bear and coyote of Tennessee.

Angiostrongylus vasorum, commonly known as the French heartworm, is a metastrongyloid parasitic nematode that infects wild and domestic canids. In North America, A. vasorum is endemic to the Canadian island of Newfoundland, but has been expanding to new areas including Nova Scotia, Prince Edward Island, and West Virginia (USA). Two cases of A. vasorum are reported from the state of Tennessee. The first case in a black bear (Ursus americanus) and the second case in a coyote (Canis latrans). The black bear was found dead in Sevier County in November of 2022, while the coyote was trapped and euthanized as part of a predator control program in Campbell County in January of 2023. Histology of the lungs revealed both animals had verminous pneumonia. DNA was extracted from the lungs of both, and PCR was performed using NC1 and NC2 primers. Sequencing results of the PCR products from the bear and coyote samples indicated that they were 95% and 96% similar, respectively, to European strains of A. vasorum. This report marks the first time A. vasorum has been reported in Tennessee as well as only the second and third report of autochthonous A. vasorum infection in the United States and the first report in an ursid. These two cases confirm the spread of A. vasorum further into North America. This nematode is highly pathogenic to wild and domestic canids, and thus these cases represent an emerging threat to both and underscore the need for further surveillance for the parasite.

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.

Abdominal angiostrongyliasis in the Americas: fifty years since the discovery of a new metastrongylid species, Angiostrongylus costaricensis.

Angiostrongylus costaricensis is a zoonotic parasitic nematode described for the first time in 1971 by Pedro Morera and Rodolfo Céspedes in Costa Rica. This parasite causes an infection known as abdominal angiostrongyliasis, affecting mainly school-aged children and young adults. Infection with A. costaricensis has been associated with a myriad of rodent and mollusk species in the Americas and the Caribbean, as its natural hosts and reservoirs. In this commemorative review, we highlight the extensive research collected through a 50-year journey, which includes ecological, pathological, and molecular studies on A. costaricensis and its implicated disease. We also identify major knowledge gaps in its evolutionary history, the ecological role of imported and invasive mollusk species, and immune response. We propose that the advent of -omics analyses will allow us to gather novel information regarding A. costaricensis biology and infection dynamics, as well as to promote the design of much-needed sensitive and specific diagnostic tools.

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.

Angiostrongylus spp. (Nematoda; Metastrongyloidea) of global public health importance.

Human angiostrongyliasis is an important foodborne zoonosis, caused by the infection with Angiostrongylus costaricensis and Angiostrongylus cantonensis. These two species have a significant public health impact in different areas of the world. Angiostrongyliasis is re-emerging and expanding to urban settings rising significant concerns regarding the control of these infections. This review focuses on aspects such as life cycle, epidemiology, clinical manifestations, diagnostics, food safety and control of illness caused especially by A. cantonensis.

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.

Two Ocular Angiostrongyliasis Cases in Thailand with Molecular Identification of Causative Parasite Species.

Human angiostrongyliasis is an important foodborne helminthic zoonosis caused by the nematodes of the genus Angiostrongylus. We describe two parasitologically confirmed cases of ocular angiostrongyliasis, presenting at a tertiary care hospital in Thailand between 2012 and 2018. Parasites were surgically recovered from patients' eyes and morphologically identified as Angiostrongylus species. DNA analysis allowed identification of Angiostrongylus cantonensis. Polymerase chain reaction was used to amplify all or part of the small nuclear ribosomal subunit, the second internal transcribed spacer region, and cytochrome c oxidase subunit 1. The sequences subsequently obtained were highly similar to those of A. cantonensis (97-100%). This is the first molecular confirmation that A. cantonensis is a causative agent of human angiostrongyliasis in Thailand.

The mitochondrial genome of Angiostrongylus mackerrasae is distinct from A. cantonensis and A. malaysiensis.

The native rat lungworm (Angiostrongylus mackerrasae) and the invasive rat lungworm (Angiostrongylus cantonensis) occur in eastern Australia. The species identity of A. mackerrasae remained unquestioned until relatively recently, when compilation of mtDNA data indicated that A. mackerrasae sensu Aghazadeh et al. (2015b) clusters within A. cantonensis based on their mitochondrial genomes (mtDNA). To re-evaluate the species identity of A. mackerrasae, we sought material that would be morphologically conspecific with A. mackerrasae. We combined morphological and molecular approaches to confirm or refute the specific status of A. mackerrasae. Nematodes conspecific with A. mackerrasae from Rattus fuscipes and Rattus rattus were collected in Queensland, Australia. Morphologically identified A. mackerrasae voucher specimens were characterized using amplification of cox1 followed by the generation of reference complete mtDNA. The morphologically distinct A. cantonensis, A. mackerrasae and A. malaysiensis are genetically distinguishable forming a monophyletic mtDNA lineage. We conclude that A. mackerrasae sensu Aghazadeh et al. (2015b) is a misidentified specimen of A. cantonensis. The availability of the mtDNA genome of A. mackerrasae enables its unequivocal genetic identification and differentiation from other Angiostrongylus species.

Angiostrongylus minasensis n. sp.: new species found parasitizing coatis (Nasua nasua) in an urban protected area in Brazil.

Currently, there are 21 species of Angiostrongylus that parasitize the pulmonary or mesenteric arteries of wild and domestic rodents, felids, canids and human. Species of Angiostrongylus have cosmopolitan distribution covering tropical, subtropical and temperate regions. The procyonid Nasua nasua (coati) is a reservoir host for a wide variety of parasites that may be harmful to its populations or may contain etiological agents with zoonotic potential. In urban areas, coatis are usually found in close association with humans and domestic animals. We morphologically and molecularly characterized a new species of Angiostrongylus found in N. nasua in a protected area within Belo Horizonte, Brazil. The new species of Angiostrongylus differs from other species of the same genus in terms of the length and bifurcation level of the lateral and ventral rays, the length of spicules and female tail morphology. Molecular phylogenetic results based on the mitochondrial cytochrome c oxidase subunit 1 gene suggest that the newly identified species belongs to a genetic lineage that is separate from other species of Angiostrongylus. This new species was collected from the mesenteric arteries of N. nasua. It was named Angiostrongylus minasensis n. sp..

Angiostrongylus minasensis n. sp.: new species found parasitizing coatis (Nasua nasua) in an urban protected area in Brazil / Angiostrongylus minasensis n. sp.: nova espécie encontrada parasitando quatis (Nasua nasua) em área de proteção urbana no Brasil

Abstract Currently, there are 21 species of Angiostrongylus that parasitize the pulmonary or mesenteric arteries of wild and domestic rodents, felids, canids and human. Species of Angiostrongylus have cosmopolitan distribution covering tropical, subtropical and temperate regions. The procyonid Nasua nasua (coati) is a reservoir host for a wide variety of parasites that may be harmful to its populations or may contain etiological agents with zoonotic potential. In urban areas, coatis are usually found in close association with humans and domestic animals. We morphologically and molecularly characterized a new species of Angiostrongylus found in N. nasua in a protected area within Belo Horizonte, Brazil. The new species of Angiostrongylus differs from other species of the same genus in terms of the length and bifurcation level of the lateral and ventral rays, the length of spicules and female tail morphology. Molecular phylogenetic results based on the mitochondrial cytochrome c oxidase subunit 1 gene suggest that the newly identified species belongs to a genetic lineage that is separate from other species of Angiostrongylus. This new species was collected from the mesenteric arteries of N. nasua. It was named Angiostrongylus minasensis n. sp..

Resumo Existem 21 espécies de Angiostrongylus que parasitam as artérias pulmonares ou mesentéricas de roedores silvestres e domésticos, felídeos, canídeos e homem. Espécies de Angiostrongylus têm uma distribuição cosmopolita que abrange regiões tropicais, subtropicais e temperadas. O procionídeo Nasua nasua (quati) é hospedeiro de vários parasitos que podem ser prejudiciais para suas populações ou conter agentes etiológicos com potencial zoonótico. Nas áreas urbanas, os quatis podem ser encontrados em estreita associação com seres humanos e animais domésticos. Nós caracterizamos morfológica e molecularmente uma nova espécie de Angiostrongylus encontrada em N. nasua de uma área protegida na cidade de Belo Horizonte, Brasil. A nova espécie de Angiostrongylus difere de outras espécies do mesmo gênero pelo comprimento e nível de bifurcação dos raios lateral e ventral, o comprimento dos espículos e a morfologia da cauda da fêmea. Resultados moleculares e filogenéticos baseados no gene mitocondrial citocromo c oxidase subunidade 1 indicam que a espécie recém-identificada pertence a uma linhagem genética separada de outras espécies de Angiostrongylus. O presente relato descreve uma nova espécie de Angistrongylus coletada das artérias mesentéricas de N. nasua, denominada Angiostrongylus minasensis n. sp..

Annotated catalogue of species of Angiostrongylus and the related genera Gallegostrongylus, Rodentocaulus and Stefanskostrongylus (Nematoda: Metastrongyloidea, Angiostrongylidae).

This catalogue is concerned with the closely related angiostrongylid genera Angiostrongylus, Gallegostrongylus, Rodentocaulus and Stefanskostrongylus. Three species, Angiostrongylus cantonensis, A. costaricensis and A. vasorum, have attracted most attention because of their importance in human and domestic animal disease. Many of the remaining species are poorly known and the number of valid taxa is unclear. The catalogue lists all nomenclaturally available and unavailable genus-group and species-group names that have been applied to the above genera and the species included in them, indicating their current nomenclatural status and providing the rigorous nomenclatural basis for future work. The catalogue lists 14 published and nomenclaturally available genus-group names, with the above four treated as valid, the other ten being junior synonyms. There are 42 published species-group names: 36 are valid, two are junior synonyms, four are nomenclaturally unavailable. One additional species, described in Chabaudistrongylus (synonym of Angiostrongylus), is listed as incertae sedis in Angiostrongylidae. Also listed are two unpublished collection names. The catalogue provides bibliographic details for all published names, and for available names provides locations of type material, details of type localities, geographic distributions and details of type and other hosts, both definitive and intermediate, to the extent known. The catalogue is a work of nomenclature, not a revisionary taxonomic work. No new names or new combinations are proposed. The apparently new family-group synonymy of Cardionematinae with Angiostrongylidae is introduced, as are four genus-group synonymies, three with Angiostrongylus and one with Stefaskostrongylus.

Distribution of the newly invasive New Guinea flatworm Platydemus manokwari (Platyhelminthes: Geoplanidae) in Thailand and its potential role as a paratenic host carrying Angiostrongylus malaysiensis larvae.

Invasive species constitute one of the most serious threats to biodiversity and ecosystems, and they potentially cause economic problems and impact human health. The globally invasive New Guinea flatworm, Platydemus manokwari (Platyhelminthes: Geoplanidae), has been identified as a threat to terrestrial biodiversity, particularly soil-dwelling native species (e.g. molluscs, annelids and other land planarians), and is listed among 100 of the world's worst invasive alien species. We report here, for the first time, P. manokwari occurrences in many locations throughout Thailand, using voluntary digital public participation from the social network portals associated with the Thailand Biodiversity Conservation Group and collections of living flatworm specimens. Mitochondrial cytochrome c oxidase subunit I (COI) sequences confirmed that all collected flatworms were P. manokwari and placed them in the "world haplotype" clade alongside other previously reported specimens from France, Florida (USA), Puerto Rico, Singapore, French Polynesia, New Caledonia, and the Solomon Islands. In addition, infective stage larvae (L3) of the nematode Angiostrongylus malaysiensis were found in the flatworm specimens, with a 12.4% infection rate (15/121 specimens examined). Platydemus manokwari occurrence in Thailand and its capacity to carry L3 of Angiostrongylus should be of concern to biodiversity conservation and human health practitioners, because this invasive flatworm species may be involved in the life cycle of angiostrongylid worms in Thailand.

Invasive slug Meghimatium pictum (Stoliczka, 1873) infected by Angiostrongylus costaricensis Morera & Céspedes, 1971, and the possible risk of human infection associated with grape consumption.

Many molluscs may be infected with angiostrongylid larvae. Following the histopathological diagnosis of abdominal angiostrongyliasis in a grape farmer from southern Brazil, molluscs in the area were investigated. During a nocturnal search, 245 specimens of slugs were collected and identified as the invasive Chinese slug Meghimatium pictum. Angiostrongylus costaricensis worms were recovered from mice that were experimentally infected with larvae obtained from 11 (4.5%) of the molluscs. This study presents the first report of M. pictum being identified as an intermediate host for A. costaricensis. Most of the slugs were collected from grape plants, which suggests that transmission may be associated with grape consumption.

Angiostrongylus spp. in the Americas: geographical and chronological distribution of definitive hosts versus disease reports

BACKGROUND Angiostrongyliasis is an infection caused by nematode worms of the genus Angiostrongylus. The adult worms inhabit the pulmonary arteries, heart, bronchioles of the lung, or mesenteric arteries of the caecum of definitive host. Of a total of 23 species of Angiostrongylus cited worldwide, only nine were registered in the American Continent. Two species, A. cantonensis and A. costaricensis, are considered zoonoses when the larvae accidentally parasitise man. OBJECTIVES In the present study, geographical and chronological distribution of definitive hosts of Angiostrongylus in the Americas is analysed in order to observe their relationship with disease reports. Moreover, the role of different definitive hosts as sentinels and dispersers of infective stages is discussed. METHODS The study area includes the Americas. First records of Angiostrongylus spp. in definitive or accidental hosts were compiled from the literature. Data were included in tables and figures and were matched to geographic information systems (GIS). FINDINGS Most geographical records of Angiostrongylus spp. both for definitive and accidental hosts belong to tropical areas, mainly equatorial zone. In relation to those species of human health importance, as A. cantonensis and A. costaricensis, most disease cases indicate a coincidence between the finding of definitive host and disease record. However, in some geographic site there are gaps between report of definitive host and disease record. In many areas, human populations have invaded natural environments and their socioeconomic conditions do not allow adequate medical care. MAIN CONCLUSIONS Consequently, many cases for angiostrongyliasis could have gone unreported or unrecognised throughout history and in the nowadays. Moreover, the population expansion and the climatic changes invite to make broader and more complete range of observation on the species that involve possible epidemiological risks. This paper integrates and shows the current distribution of Angiostrongylus species in America, being this information very relevant for establishing prevention, monitoring and contingency strategies in the region.

Angiostrongylus spp. in the Americas: geographical and chronological distribution of definitive hosts versus disease reports.

BACKGROUND: Angiostrongyliasis is an infection caused by nematode worms of the genus Angiostrongylus. The adult worms inhabit the pulmonary arteries, heart, bronchioles of the lung, or mesenteric arteries of the caecum of definitive host. Of a total of 23 species of Angiostrongylus cited worldwide, only nine were registered in the American Continent. Two species, A. cantonensis and A. costaricensis, are considered zoonoses when the larvae accidentally parasitise man. OBJECTIVES: In the present study, geographical and chronological distribution of definitive hosts of Angiostrongylus in the Americas is analysed in order to observe their relationship with disease reports. Moreover, the role of different definitive hosts as sentinels and dispersers of infective stages is discussed. METHODS: The study area includes the Americas. First records of Angiostrongylus spp. in definitive or accidental hosts were compiled from the literature. Data were included in tables and figures and were matched to geographic information systems (GIS). FINDINGS: Most geographical records of Angiostrongylus spp. both for definitive and accidental hosts belong to tropical areas, mainly equatorial zone. In relation to those species of human health importance, as A. cantonensis and A. costaricensis, most disease cases indicate a coincidence between the finding of definitive host and disease record. However, in some geographic site there are gaps between report of definitive host and disease record. In many areas, human populations have invaded natural environments and their socioeconomic conditions do not allow adequate medical care. MAIN CONCLUSIONS: Consequently, many cases for angiostrongyliasis could have gone unreported or unrecognised throughout history and in the nowadays. Moreover, the population expansion and the climatic changes invite to make broader and more complete range of observation on the species that involve possible epidemiological risks. This paper integrates and shows the current distribution of Angiostrongylus species in America, being this information very relevant for establishing prevention, monitoring and contingency strategies in the region.

Cytochrome c oxidase subunit I haplotype reveals high genetic diversity of Angiostrongylus malaysiensis (Nematoda: Angiostrongylidae).

The rat lungworm Angiostrongylus malaysiensis is a metastrongyloid nematode parasite. It has been reported in Malaysia, Thailand, Laos, Myanmar, Indonesia and Japan. In this study, A. malaysiensis adult worms recovered from the lungs of wild rats in different geographical regions/provinces in Thailand were used to determine their haplotype by means of the mitochondrial partial cytochrome c oxidase subunit I (COI) gene sequence. The results revealed high COI haplotype diversity of A. malaysiensis from Thailand. The geographical isolates of A. malaysiensis from Thailand and other countries formed a monophyletic clade distinct from the closely related A. cantonensis. In the present study, five new haplotypes were identified in addition to the four haplotypes reported in the literature. Phylogenetic analysis revealed that four of these five new haplotypes - one from Mae Hong Song (northern region), two from Tak (western region) and one from Phang Nga (southern region) - formed a distinct clade with those from Phatthalung (southern region) and Malaysia. The haplotype from Malaysia was identical to that of Phatthalung (haplotype AM1). In general, the COI sequences did not differentiate unambiguously the various geographical isolates of A. malaysiensis. This study has confirmed the presence of high COI genetic diversity in various geographical isolates of A. malaysiensis. The COI gene sequence will be suitable for studying genetic diversity, population structure and phylogeography.

Angiostrongylus mackerrasae and A. cantonensis (Nematoda: Metastrongyloidea) belong to same genetic lineage: evidence from mitochondrial protein-coding genes.

Angiostrongylus mackerrasae is a parasitic nematode of rats found in Australia. When first reported, it was referred to as A. cantonensis. Recent molecular studies, including the mitochondrial genome, indicate that it is highly similar to A. cantonensis. These studies did not include A. malaysiensis, another member of the A. cantonensis species complex, for comparison. The present study examined the genetic distance and phylogenetic relationship between the component taxa (A. cantonensis, A. mackerrasae and A. malaysiensis) of the A. cantonensis species complex, based on the 12 protein-coding genes (PCGs) of their mitochondrial genome. Both the nucleotide and amino acid sequences were analysed. Angiostrongylus mackerrasae and A. cantonensis are members of the same genetic lineage and both are genetically distinct from A. malaysiensis. The genetic distance based on concatenated nucleotide sequences of 12 mt-PCGs between A. mackerrasae and A. cantonensis from Thailand is p = 1.73%, while that between the Thai and Chinese taxa of A. cantonensis is p = 3.52%; the genetic distance between A. mackerrasae and A. cantonensis from China is p = 3.70%. The results indicate that A. mackerrasae and A. cantonensis belong to the same genetic lineage, and that A. mackerrasae may be conspecific with A. cantonensis. It remains to be resolved whether A. mackerrasae is conspecific with A. cantonensis or undergoing incipient speciation.

A rare cardiopulmonary parasite of the European badger, Meles meles: first description of the larvae, ultrastructure, pathological changes and molecular identification of Angiostrongylus daskalovi Janchev & Genov 1988.

BACKGROUND: Angiostrongylus daskalovi is a rare cardiopulmonary nematode infecting badgers. The parasite was described in 1988 and, since then, found only once in mustelids in Europe. The present study aims to report new cases of patent A. daskalovi infection in badgers from northern Romania and to provide new information on its ultrastructure, molecular diagnosis, and pathology. METHODS: Eight road-killed or hunted badgers originating from Maramureș and Alba counties in Romania were collected and necropsied. Adults and larvae of cardio-pulmonary nematodes were collected and examined by light and scanning electron microscopy (SEM). Genomic DNA was extracted from adults and first-stage larvae (L1). PCR amplification of the internal transcribed spacer 2 (ITS2, ∼500 bp) of the rRNA gene was performed. Amplicons were purified, sequenced, and compared to those available in the GenBank database. Histopathological examination of the lungs was performed and lesions described. RESULTS: The necropsy revealed the presence of nematodes in the pulmonary arteries of three animals. All parasites were mature adults and the coproscopic examination showed the presence of eggs and L1 larvae in all three positive animals. Light microscopy examination confirmed the morphological and morphometric similarity of parasites to Angiostrongylus daskalovi. SEM highlighted the typical angiostrongylid structure of the rays of the copulatory bursa and the anterior extremity, with the presence of six sensory papillae surrounding the mouth opening in which a triangular tooth was visible. The first-stage larva (L1) of A. daskalovi is described here for the first time. Histopathological examination of the lungs showed chronic interstitial verminous pneumonia due to the presence of adult parasites. Molecular analysis showed 100 % nucleotide similarity to an Angiostrongylus sp. isolate originating from a badger from Spain, tentatively identified as A. daskalovi. CONCLUSIONS: Our study unequivocally demonstrates the presence of A. daskalovi in European badgers from Romania, provides the first description of the larvae and reveals new data about the ultrastructure of adult parasites and their pathological impact, contributing to the understanding of the phylogenetic relationships with other congeneric species.

Complete mitochondrial genome of Angiostrongylus malaysiensis lungworm and molecular phylogeny of Metastrongyloid nematodes.

Angiostrongylus malaysiensis is a nematode parasite of various rat species. When first documented in Malaysia, it was referred to as A. cantonensis. Unlike A. cantonensis, the complete mitochondrial genome of A. malaysiensis has not been documented. We report here its complete mitogenome, its differentiation from A. cantonensis, and the phylogenetic relationships with its congeners and other Metastrongyloid taxa. The whole mitogenome of A. malaysiensis had a total length of 13,516bp, comprising 36 genes (12 PCGs, 2 rRNA and 22 tRNA genes) and a control region. It is longer than that of A. cantonensis (13,509bp). Its control region had a long poly T-stretch of 12bp which was not present in A. cantonensis. A. malaysiensis and A. cantonensis had identical start codon for the 12 PCGs, but four PCGs (atp6, cob, nad2, nad6) had different stop codon. The cloverleaf structure for the 22 tRNAs was similar in A. malaysiensis and A. cantonensis except the TΨC-arm was absent in trnV for A. malaysiensis but present in A. cantonensis. The Angiostrongylus genus was monophyletic, with A. malaysiensis and A. cantonensis forming a distinct lineage from that of A. costaricensis and A. vasorum. The genetic distance between A. malaysiensis and A. cantonensis was p=11.9% based on 12 PCGs, p=9.5% based on 2 rRNA genes, and p=11.6% based on 14 mt-genes. The mitogenome will prove useful for studies on phylogenetics and systematics of Angiostrongylus lungworms and other Metastrongyloid nematodes.