Occurrence and identification of lungworms in Iranian wild boars.

This study represents the first investigation into the occurrence and identification of Metastrongylus spp. in wild boars (Sus scrofa) in Iran, utilizing both molecular and morphological methods. Thirteen wild boars from Kerman Province were examined, with 92.3% found to be infected with at least one species of Metastrongylus. Mixed infections were observed in 38.46% of the animals. Morphological and molecular analyses confirmed the presence of M. pudendotectus and M. salmi, with prevalence rates of 76.9% and 53.9%, respectively. Histopathological examination revealed transverse and longitudinal sections of Metastrongylus parasites within the airways, causing partial to complete obstruction, interstitial pneumonia, and inflammatory responses. The study also highlights the public health significance of these parasites. The higher prevalence observed compared to earlier studies suggests changes in environmental conditions, host dynamics, or agricultural practices as possible factors, warranting further investigation. The findings underscore the need for comprehensive surveillance and control measures to mitigate the risk of zoonotic transmission, particularly in regions with significant wild and domestic swine populations. This study contributes to the understanding of Metastrongylus spp. distribution and their pathological impact, emphasizing the ecological importance of wild boars and the necessity for continued monitoring and research to prevent and control infections in both animal and human populations.

Expression profiles of miRNAs in the lung tissue of piglets infected with Glaesserella parasuis and the roles of ssc-miR-135 and ssc-miR-155-3p in the regulation of inflammation.

MicroRNAs (miRNAs) have been shown to play an important regulatory role in the process of pathogenic infection. However, the miRNAs that regulate the pathogenic process of G. parasuis and their functions are still unknown. Here, high-throughput sequencing was used to quantify the expression of miRNA in piglet lung tissue after G. parasuis XX0306 strain infection. A total of 25 differentially expressed microRNAs (DEmiRNAs) were identified. GO and KEGG pathway enrichment analysis showed that many of the functions of genes that may be regulated by DEmiRNA are related to inflammatory response and immune regulation. Further studies found that ssc-miR-135 may promote the expression of inflammatory factors through NF-κB signaling pathway. Whereas, ssc-miR-155-3p inhibited the inflammatory response induced by G. parasuis, and its regulatory mechanism remains to be further investigated. This study provides a valuable reference for revealing the regulatory effects of miRNAs on the pathogenesis of G. parasuis. DATA AVAILABILITY: The datasets generated during the current study are not publicly available due to this study is currently in the ongoing research stage, and some of the data cannot be made public sooner yet, but are available from the corresponding author on reasonable request.

Molecular prevalence, phylogenetic characterization, and epidemiological risk factors of pulmonary nematodes in domestic cats in Türkiye.

Aelurostrongylus abstrusus, Oslerus rostratus, and Troglostrongylus brevior are nematodes found in the respiratory system of domestic cats and cause a variety of symptoms. All three parasites use the same hosts and niches, and the morphological measurements of their L1s excreted in faeces overlap with each other. In this study, 300 cats brought to Ondokuz Mayis University Veterinary Teaching Animal Hospital were screened for lungworms by morphological measurements and molecular analyses. The prevalence of the lungworms was found as 1.33% (4/300) for A. abstrusus, 0.66% (2/300) for O. rostratus, and 0.33% (1/300) for T. brevior. Molecular identification of A. abstrusus, T. brevior, and O. rostratus in domestic cats was carried out for the first time in Türkiye within the present study. This study also reveals the risk factors of lungworm positivity in domestic cats in Türkiye.

First report of Orthostrongylus sp. (Nematoda: Protostrongylidae) in wild reindeer (Rangifer tarandus) from the Taimyr, Russia: Nearctic parasites in a Palearctic host.

First stage larvae of an unknown lungworm (Protostrongylidae) were isolated in the feces of a wild reindeer (Rangifer tarandus) from Taimyr, Russia. Larvae were 365-366 µm long and had a tail spike lacking a dorsal spine. DNA analyses using BLAST showed that nuclear sequences obtained (LSU rDNA, 825 bp and ITS2 rDNA, 395 bp) were highly similar (99.50% and 98.88% identity, respectively) to an isolate of Orthostrongylus macrotis (GenBank: EU595592.1) from North America. It cannot be confirmed whether these larvae represent an uncharacterized species of Orthostrongylus or can be referred to O. macrotis, a species that has historically only been reported from the Nearctic. This is the first report of lungworms attributable to Protostrongylinae in R. tarandus across its vast geographic in the Holarctic.

Cardio-pulmonary parasites of the European wildcat (Felis silvestris) in Germany.

BACKGROUND: In the last years, research on feline cardio-pulmonary parasites has considerably increased in Europe. Not only domestic cats (Felis catus), but also European wildcats (Felis silvestris) can serve as definitive hosts for these nematodes. The F. silvestris population in Germany has been growing rapidly within the last decades; therefore, the assessment of its cardio-pulmonary parasite status is of importance to unravel whether the wildcat population serves as a substantial reservoir for these nematodes and might pose a health threat to domestic cats. METHODS: As part of a nature conservation project for European wildcats in the German federal state Rhineland-Palatinate, lungs (n = 128) and hearts (n = 111) of 128 F. silvestris found dead were examined for cardio-pulmonary parasites. All isolated parasites were identified morphologically, and results were confirmed by molecular analysis of a total of 3-11 specimens of each worm species. RESULTS: A total of 70.3% (90/128) wildcats were positive for at least one lungworm species. Angiostrongylus chabaudi was most common (53.1% [68/128]), followed by Aelurostrongylus abstrusus (42.2% [54/128]), Troglostrongylus brevior (31.3% [40/128]) and Capillaria spp. (3.1% [4/128]). Of note, about two-thirds of the infected wildcats harboured coinfections. Infection intensities ranged from 1 to 167 nematodes per wildcat. Generalised linear models revealed a strong correlation between A. chabaudi and A. abstrusus infection, and prevalences were higher in adult than in younger wildcats, except for T. brevior. Moreover, the T. brevior prevalence varied significantly with nutritional status. CONCLUSIONS: This study shows that feline cardio-pulmonary nematodes are common parasites in European wildcats in Germany but do not appear to have a serious impact on the overall health of the population. Due to presumed spillover events via prey, cardio-pulmonary nematodes may circulate between the wildcat population and domestic cats and might therefore pose a health risk to individual domestic cats.

Comprehensive Molecular Characterization of the Mitochondrial Genome of the Takin Lungworm Varestrongylus eleguneniensis (Strongylida: Protostrongylidae).

The takin lungworm Varestrongylus eleguneniensis (Strongylida: Protostrongylidae) causes lethal bronchopneumonia and represents severe threats to captive and wild populations. However, until now there has been very limited information available concerning the molecular epidemiology and evolutionary biology of V. eleguneniensis. Mitochondrial genomes (mtDNAs) can provide resources for investigations in these areas and, therefore, can assist with the surveillance and control of this lungworm. Herein, the complete mtDNA of V. eleguneniensis was sequenced and characterized with Illumina pipeline analyses. This circular genome (13,625 bp) encoded twelve protein-coding genes (PCGs), two rRNAs, and twenty-two tRNAs, with notable levels of AT and GC skews. Comparative genomics revealed a purifying selection among PCGs, with cox1 and nad6 having the lowest and the highest evolutionary rate, respectively. Genome-wide phylogenies showed a close relationship between V. eleguneniensis and Protostrongylus rufescens in Strongylida. Single gene (PCGs or rRNAs)-based phylogenies indicated that cox1 and nad5 genes shared the same family-level topology with that inferred from genomic datasets, suggesting that both genes could be suitable genetic markers for evolutionary and phylogenetic studies of Strongylida species. This was the first mtDNA of any member of the genus Varestrongylus, and its comprehensive molecular characterization represents a new resource for systematic, population genetic and evolutionary biological studies of Varestrongylus lungworms in wildlife.

Molecular detection of Metastrongylus salmi eggs from pigs in low-resource communities in the state of Piauí, northeastern Brazil.

Metastrongylosis is an infection of the respiratory tract of pigs caused by parasites of the genus Metastrongylus, whose eggs are similar to other Strongylida through light microscopy; species-specific identification can be performed with molecular tools. We explored the species composition and the genetic diversity of Metastrongylus infecting pigs in close contact with humans in impoverished rural communities in the state of Piauí, in northeastern Brazil. Fecal samples (n = 78) were collected for parasitologic tests. Egg morphometry and molecular characterization, using the cytochrome c oxidase subunit 1 (cox1) gene, were performed. For strongyliform eggs, 62 of 78 (80%) pigs were positive and 6 of 99 (6%) eggs had dimensions compatible with Metastrongylus. Of the 37 samples submitted to PCR, 10 were identified as M. salmi. We found 3 M. salmi haplotypes, including 2 new and 1 described previously in Europe. Overall, M. salmi demonstrated lower intraspecific genetic diversity: diversity index (H) ± SD = 0.318 ± 0.164, n = 12, compared with published M. pudendotectus sequences (1.000 ± 0.272, n = 3). To our knowledge, M. salmi DNA sequences have not been published previously from pigs in South America.

Molecular phylogenetic relationships of Metastrongylus nematodes with emphasis on specimens from domestic pigs in Vietnam.

Lungworms of the genus Metastrongylus are parasitic nematodes in the respiratory tract of swine. Although they infect both wild boars and domestic pigs, studies on Metastrongylus infections in wild boars in Europe, the Americas and Africa are numerous, while those in domestic pigs are few. There are several studies analysing the molecular phylogenetic relationships of few individual Metastrongylus species with other nematode taxa, but there are no studies on the phylogenetic relationships of species within the genus Metastrongylus. In Southeast Asia, reports on swine lungworms are extremely scarce and do not include any nucleotide sequence data. Therefore, the aim of the present study is to survey Metastrongylus infection in domestic pigs raised in Dien Bien Province, Northern Vietnam, and to analyse the molecular phylogenetic relationships of Metastrongylus species. Based on morphological and molecular data, we identified two species: Metastrongylus apri and Metastrongylus pudendotectus. The prevalence of the former species was found to be significantly higher than the latter one (24.1% vs. 2.3%). We observed pigs exhibiting a coinfection with the two lungworm species or a single infection with only M. apri. However, we did not observe any pigs being infected with just M. pudendotectus. Vietnamese Metastrongylus specimens showed slight morphological and molecular differences compared to those from other countries. The molecular analyses revealed a close genetic relationship between M. apri and Metastrongylus salmi, while both these species were far distant from M. pudendotectus. The present study highlights the needs for further studies to clarify the morphological features and ecological and phylogenetic relationships of Metastrongylus species at the global scale.

Pathologic features and molecular identification of parelaphostrongylosis in a sitatunga (Tragelaphus spekii).

We report the pathologic features, local inflammatory response immunophenotype, and molecular identification results of cerebral nematodiasis in a young sitatunga (Tragelaphus spekii) from Texas. To the authors' knowledge, this is the first report of cerebral nematodiasis by Parelaphostrongylus tenuis in a sitatunga, a bovid species introduced into the USA, and the first characterization of the local inflammatory response immunoprofile in this condition. A molecular identification method based on formalin-fixed and paraffin-embedded-polymerase chain reaction was described. These results contribute to knowledge on geographical distribution and host spectrum of P. tenuis, and highlight the relevance of this nematodiasis in naïve translocated or introduced bovid species into endemic areas.

Determinants of lungworm specificity in five cetacean species in the western Mediterranean.

BACKGROUND: Current data about Pseudaliidae show contrasting patterns of host specificity between congeneric species. We investigated how both contact and compatibility between hosts and parasites contributed to the patterns of lungworm infection observed in a community of five species of cetaceans in the western Mediterranean. METHODS: The lungs of 119 striped dolphins Stenella coeruleoalba, 18 bottlenose dolphins Tursiops truncatus, 7 Risso's dolphins Grampus griseus, 7 long-finned pilot whales Globicephala melas, and 6 common dolphins Delphinus delphis were analysed for lungworms. Parasites were identified by morphology and analysis of ITS2 sequences using both maximum likelihood and Bayesian inference methods. Body length was used as a proxy for lungworm species fitness in different hosts and compared with Kruskal-Wallis tests. Infection parameters were compared between cetacean species using Fisher's exact tests and Kruskal-Wallis tests. Phylogenetic specificity was explored by collating the overall lungworm species prevalence values in hosts from previous surveys in various localities. To explore the relative importance of vertical and horizontal transmission, Spearman's rank correlation was used to look for an association between host size and lungworm burden. A Mantel test was used to explore the association between lungworm species similarity and prey overlap using dietary data. RESULTS: Halocercus delphini had higher infection levels in striped dolphins and common dolphins; Stenurus ovatus had higher infection levels in bottlenose dolphins; and Stenurus globicephalae had higher infection levels in long-finned pilot whales. These results are congruent with findings on a global scale. Morphometric comparison showed that the larger nematodes were found in the same host species that had the highest parasite burden. Lungworms were found in neonatal striped dolphins and a Risso's dolphin, and there was a weak but significant correlation between host size and parasite burden in striped dolphins and bottlenose dolphins. There was also a weak but significant association between prey overlap and lungworm species similarity. CONCLUSIONS: Data indicate that phylogenetic specificity has an important role in governing host-parasite associations, as indicated by the higher infection levels and larger nematode size in certain hosts. However, diet can also influence infection patterns in these preferred hosts and contribute to less severe infections in other hosts.

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.

Troglostrongylus brevior is the dominant lungworm infecting feral cats in Jerusalem.

Feline lungworms infect the respiratory tract of wild and domestic cats, causing infection often associated with clinical disease. Until recently, Aelurostrongylus abstrusus has been considered the most relevant species of lungworm, while Troglostrongylus brevior was considered of less significance. Fecal samples of feral cats from Jerusalem, Israel, collected over a year, were examined for first stage lungworm larvae (L1) using the Baermann method. Positive samples were morphologically identified, and their species identity was molecularly confirmed. Forty of 400 (10.0%) cats were lungworm-positive, of which 38/40 (95.0%) shed Troglostrongylus brevior and 6/40 (15.0%) shed Aelurostrongylus abstrusus. Four cats (10.0%) had mixed infections with both lungworm species. L1 shedding was associated with clinical respiratory signs in 11 (19.0%) T. brevior shedding cats of a total of 58 cats manifesting respiratory signs, while 23/342 (6.7%) cats without respiratory signs were L1-positive (p = 0.006). Non-respiratory clinical signs were also found to be more prevalent in L1 shedders (p = 0.012). A young kitten ≤ 4 weeks of age shed T. brevior L1 larvae. DNA sequences of both lungworm species using the ribosomal internal transcribed spacer 2 (ITS2) locus were > 99% similar to other sequences deposited in GenBank, suggesting that T. brevior and A. abstrusus ITS2 sequences are both highly conserved. In conclusion, L1 shedding in feral cats from Jerusalem were mostly caused by T. brevior with only a small proportion involving A. abstrusus, different from many studies from other geographical regions.

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.

Surprisingly long body length of the lungworm Parafilaroides gymnurus from common seals of the Dutch North Sea.

Lungworms of the genera Parafilaroides and Otostrongylus are responsible for parasitic bronchopneumonia, the foremost disease of eastern Atlantic common seals (EACS, Phoca vitulina vitulina) in the Dutch North Sea. Recently, there have been increased reports of lungworm cases and observations of unusually long Parafilaroides sp. adults in this location. The initial aim of this study was to confirm the identity of the Parafilaroides species infecting this population. Parafilaroides are usually small and delicate, making them difficult to extract from host tissue, and there is often difficulty accessing fresh specimens for morphological study. The large size of the Dutch worms and the accessibility of specimens from numerous animals enabled the description and measurement of many intact specimens (N = 64) from multiple host animals (N = 20). Species identity was confirmed by targeted sequencing of ribosomal and mitochondrial DNA amplicons from a subset of worms. Worm morphology was consistent with descriptions for P. gymnurus, but the mature females were 1.9-fold and 3.4-fold longer than those recovered from French EACS (P ≤ 0.001) and Canadian western Atlantic common seals (Phoca vitulina concolor; P ≤ 0.0001). They were also significantly longer than mature female P. gymnurus described from other seal species, with the exception of those from harp seals of Les Escoumins, Quebec. We suggest that intraspecific genetic differences in P. gymnurus and the environment within the host could contribute to the variation reported here. This study is the first to describe P. gymnurus using morphological and molecular methods and should serve as a reference for identification of the species.

Occurrence of canine and feline lungworms in Arion vulgaris in a park of Vienna: First report of autochthonous Angiostrongylus vasorum, Aelurostrongylus abstrusus and Troglostrongylus brevior in Austria.

So far, neither the feline lungworms Aelurostrongylus abstrusus and Troglostrongylus brevior nor the canine lungworm Angiostrongylus vasorum was reported in wildlife or intermediate hosts from Austria. The slug Arion vulgaris represents an invasive species in Europe and serves as intermediate host for several lungworm species. This study aimed to analyse the occurrence of metastrongyloid lungworm larvae in slugs in Vienna, Austria. Therefore, 193 A. vulgaris were collected in the central Prater park in summer 2016. Specimens were artificially digested, analysed microscopically for lungworm larvae, and species were confirmed via PCR and sequencing. Out of 193, five slugs were positive to lungworms (2.6%), one for A. vasorum, two for A. abstrusus (genotypes A and B) and one for T. brevior, and one slug had a mixed infection of A. abstrusus and T. brevior larvae. The current study is the first evidence on the endemicity of these metastrongyloid lungworm species in Austria.

Lungworm (Crenosoma goblei) Infection in Unweaned Free-Ranging Raccoons (Procyon lotor) in Washington State, USA.

Larval parasites morphologically identified as Crenosoma goblei were found in orphaned raccoons (Procyon lotor) admitted to a wildlife rehabilitation center in Washington State, US. Partial 18S rRNA and cytochrome oxidase I gene sequences were most similar to other Crenosoma spp. Although commonly reported in free-ranging raccoons throughout much of the central and eastern US, this finding extends the range of C. goblei into the Pacific Northwest of North America. Fecal samples taken within 48 h of admission were positive for larvae in 70% (21/30) of individuals and in all five group samples taken from an additional 14 raccoons. Typically, Crenosoma spp. use an intermediate host, but the unweaned status of all the raccoons in this report suggests a potential direct, possibly vertical, route of transmission. Serial fecal analysis of pooled samples collected over several subsequent months demonstrated spontaneous resolution of larval shedding. Adult worms and gross evidence of pneumonia were detected in the lungs of several juvenile raccoons at necropsy, and verminous pneumonia was confirmed histologically in the single case submitted. Clinical signs directly attributable to verminous pneumonia were not seen in most actively shedding raccoons, suggesting that subclinical disease among juveniles may be common.

First molecular identification of Aelurostrongylus abstrusus in a cat presenting severe respiratory disease from Israel.

Feline lung worm infection is increasingly reported in recent years, and recognized as a cause for respiratory disease in cats. Aelurostrongylus abstrusus is regarded as the most prevalent cause of such cases. Infective L3 larvae carried in gastropods and paratenic hosts infect felines, developing to adult worms that reside in the lungs' parenchyma and may cause verminous pneumonia. The L1 larvae hatch from eggs deposited in the lung, and are released to the environment by either feces or sputum. While the majority of epidemiological information regarding A. abstrusus originates in European countries, recent studies have shown that it is also found around the Mediterranean basin, as far east as Turkey and Cyprus. A local domestic cat from Israel showing signs of respiratory illness was diagnosed with aelurostrongylosis, confirmed by both morphological and molecular tools. Presence in Israel of this nematode was previously reported in 1949, with no further mentions since. ITS-2 sequence of the isolated larvae was highly similar to that of A. abstrusus from domestic cats from Italy. These findings show that distribution of A. abstrusus stretch to the eastern shores of the Mediterranean, and that this nematode should be considered as a cause for respiratory disease in cats in Israel and the surrounding countries.

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.

Shedding of feline lungworm larvae and their infectivity to snail intermediate hosts after anthelmintic treatment.

Aelurostrongylus abstrusus and Troglostrongylus brevior are snail-transmitted helminths causing respiratory diseases in infected cats. The shedding of feline lungworm L1s and their infectivity to the snail intermediate host, after administration of anthelminthic products to cats, are poorly documented. To assess the efficacy of 8.3% fipronil, 10% (S)-methoprene, 0.4% eprinomectin and 8.3% praziquantel (i.e. eprinomectin formulation) and 10% imidacloprid/1% moxidectin (i.e. moxidectin formulation) against these nematodes and to determine the number of days post-treatment until viable L1s are released in the faeces, 384 animals were screened by faecal examination. Of the 54 positive animals (i.e., 14.1%; 7.3% A. abstrusus, 6.2% T. brevior and 0.5% coinfected), 36 were randomly allocated to four groups. Groups A and B were composed of cats positive for T. brevior and treated with the eprinomectin and with the moxidectin formulations, respectively, whereas cats in groups C and D were positive to A. abstrusus and treated with the eprinomectin and the moxidectin formulations, respectively. Prior to and every day after treatment, faecal samples were analysed by the Baermann technique and the number of larvae per gram of faeces determined, and again four weeks after treatment, to assess the efficacy of a single administration of the products. In addition, to evaluate the pre- and post-treatment infectivity of L1s to snail intermediate hosts, one/two snails per cat were infected with 100 L1s collected from the faeces of enrolled animals and then digested 28 days p.i. Based on L1s faecal counts, the efficacy of the eprinomectin and the moxidectin formulations at 28 days was 100% for both A. abstrusus and T. brevior, with a mean number of days of 7.9 ±â€¯1.2 in group A, 7.8 ±â€¯1.9 in B, 6.9 ±â€¯1.6 in C and 8.9 ±â€¯2.0 in D to become negative. Following the artificial digestion, active L3s of T. brevior and A. abstrusus were found in 160 (87.4%) experimentally infected snails. The results of this study demonstrate that a single administration of the two formulations is effective in the treatment of A. abstrusus and T. brevior infections and that during the post-treatment period live L1s are shed for up to 8.9 ±â€¯2.0 days. L1s of both lungworm species released in the faeces after drug administration are still able to reach the infective larval stage in the infected snails. Hence, preventative measures after the treatment of infected animals should include keeping cats indoors and disposal of their faeces for approximately 10 days to avoid environmental contamination and infection of gastropod intermediate hosts.

Validity of genus Perostrongylus Schlegel, 1934 with new data on Perostrongylus falciformis (Schlegel, 1933) in European badgers, Meles meles (Linnaeus, 1758): distribution, life-cycle and pathology.

BACKGROUND: A century of debates on the taxonomy of members of the Metastrongyloidea Molin, 1861 led to many reclassifications. Considering the inconstant genus assignation and lack of genetic data, the main aim of this study was to support the validity of the genus Perostrongylus Schlegel, 1934, previously considered a synonym of Aelurostrongylus Cameron, 1927, based on new molecular phylogenetic data and to understand its evolutionary relationships with other metastrongyloid nematodes. RESULTS: Specimens of lungworm collected from European badgers in Germany, Romania and Bosnia and Herzegovina were morphologically and molecularly (rDNA, cox1) characterized. From a phylogenetic standpoint, Perostrongylus is grouped with high support together with the genera Filaroides van Beneden, 1858 and Parafilaroides Dougherty, 1946 and includes probably two species: Perostrongylus falciformis (Schlegel, 1933), a parasite of Meles meles in Europe and P. pridhami (Anderson, 1962), a parasite of Neovison vison in North America. Perostrongylus and Aelurostrongylus are assigned to different clades. Aelurostrongylus becomes a monotypic genus, with the only species Aelurostrongylus abstrusus (Railliet, 1898). In addition, we provide morphological and morphometric data for the first-stage (L1), second-stage (L2), and third-stage (L3) larvae of P. falciformis and describe their development in experimentally infected Cornu aspersum snails. The pathological and histopathological lesions in lungs of infected European badgers are also described. This is the first record of P. falciformis in Romania. CONCLUSIONS: Molecular phylogenetic and morphological data support the validity of the genus Perostrongylus, most probably with two species, P. falciformis in European badgers and P. pridhami in minks in North America. The two genera clearly belong to two different clades: Perostrongylus is grouped together with the genera Filaroides and Parafilaroides (both in the family Filaroididae Schulz, 1951), whereas Aelurostrongylus belongs to a clade with no sister groups.