Feline lungworms in Greece: copromicroscopic, molecular and serological study.

Aelurostrongylus abstrusus (Nematoda, Metastrongyloidea) causes verminous pneumonia in cats worldwide. This study evaluated the seroprevalence of A. abstrusus antibodies in 220 stray and free-roaming cats from insular (Mykonos, Crete, Skopelos) and continental (Thessaloniki, Attica) Greece. The results were compared with morphological and biomolecular identification of first-stage larvae (L1) in faeces. Positive cats were observed in all 5 areas: 13/97 (13.4%), 1/32 (3.1%), 7/26 (26.9%), 3/18 (16.7%) and 5/47 (10.6%) cats tested positive for A. abstrusus L1 by Baermann examination, and 33/97 (34.0%), 7/32 (21.9%), 6/26 (23.1%), 3/18 (16.7%) and 11/47 (23.4%) were seropositive, in Mykonos, Crete, Skopelos, Thessaloniki and Attica, respectively. Troglostrongylus brevior L1 were found in 12/97 (12.4%), 3/26 (11.5%) and 2/47 (4.3%) cats from Mykonos, Skopelos and Attica respectively. Six of the 220 cats (2.7%), i.e. 4/97 (4.1%) from Mykonos and 2/26 (7.7%) from Skopelos, shed L1 of both A. abstrusus and T. brevior. Sixty samples were ELISA-positive (27.3%, 95% CI: 21.5-33.7%), of which 21 (35%) tested copromicroscopically positive (19 monospecific infections and 2 mixed with Troglostrongylus brevior), and 5 were positive for T. brevior L1 only. Among seronegative cats (n = 140), L1 of A. abstrusus were additionally detected in 8 (5.7% out of 140) cats (i.e. 4 monospecific infections and 4 mixed with T. brevior), and in 6 (4.3% out of 140) cats, L1 of T. brevior as monospecific infection were detected. This study confirms the presence of lungworms in Greece and suggests that the number of cats infected with/exposed to metastrongylids is higher than detected by faecal examinations.

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.

Simultaneous infection by four feline lungworm species and implications for the diagnosis.

Besides Aelurostrongylus abstrusus, other parasites belonging to the superfamily Metastrongyloidea, namely Oslerus rostratus, Troglostrongylus brevior and to the family Trichuridae, i.e. Eucoleus aerophilus (syn. Capillaria aerophila), have also been reported as agents of respiratory infection in domestic cats. A case of simultaneous infection by four feline lungworm species in Sardinia is herein described. An adult female cat (Felis silvestris catus), road-killed in the southeast part of Sardinia (municipality of Villacidro, province of Cagliari), Italy, was referred to the Laboratory of Parasitology of the Veterinary Teaching Hospital in Sassari. At necropsy, the lungs were examined and dissected under a stereomicroscope for the presence of parasites, and first-stage larvae (L1) of broncho-pulmonary nematodes were searched for in a faecal sample using the Baermann method. Parasites collected in the lungs were morphologically identified as A. abstrusus, E. aerophilus, and O. rostratus. In addition to the above species, L1s of Troglostrongylus spp. were detected at coproscopy but no adult specimen was found in the lungs. The morphological identification was confirmed by the molecular amplification and sequencing of cox1 mitochondrial gene, 18S and ITS2 ribosomal DNA. This finding stands as the first simultaneous infection by four feline lungworm species in the same animal, and as the first report of O. rostratus and E. aerophilus in Sardinia.

First report of Troglostrongylus brevior in a kitten in Greece.

The first case of a natural infestation with Troglostrongylus brevior in a kitten in Greece is described here. A ∼40-day-old stray cat was referred to a private veterinary clinic with signs of respiratory distress. First stage larvae of a metastrongyloid nematode were observed in the wet mount faecal preparation. Despite an anthelmintic treatment, the respiratory signs worsened and the kitten died 2 days later. The larvae in the faeces were identified morphologically and genetically as T. brevior. The present evidence suggests a vertical or direct infestation of the kitten and a severe pathogenic role of T. brevior in young cats. This report expands the recent published cases of troglostrongylosis in domestic cats to a wider geographical distribution and opens new questions on the apparent spreading of T. brevior from wild to domestic hosts.

Mixed infection by Aelurostrongylus abstrusus and Troglostrongylus brevior in kittens from the same litter in Italy.

Parasitic nematodes affecting the respiratory system of felids are spreading in endemic regions and emerging in previously free areas and/or hosts. This is particularly the case of the cat lungworm Aelurostrongylus abstrusus, which can cause respiratory signs in cats all over the world. Additionally, Troglostrongylus brevior has been recently found in domestic cats from Ibiza Island in Spain and Southern Italy. The present paper describes the first mixed infection by these lungworms in kittens belonging to the same litter. Two ∼10-11 weeks old kittens were found infected by A. abstrusus and T. brevior at a copromicroscopical examination. The identity of larvae shed by faeces were confirmed with an already validated PCR specific for A. abstrusus and a novel DNA-based assay specific for T. brevior. One kitten showed severe respiratory signs and died despite an anthelmintic treatment, while the other had a subclinical infection and recovered after a parasiticide administration with milbemycin oxime. New insights into epidemiology, biology, clinical aspects and control of these parasitoses are discussed.

Feline lungworm Oslerus rostratus (Strongylida: Filaridae) in Italy: first case report and histopathological findings.

Oslerus rostratus syn. Anafilaroides rostratus (Strongylida: Filaroididae) is a metastrongyloid transmitted by snails, which localizes in peri-bronchial tissues and in the lung parenchyma of wild as well as domestic cats. In Europe, this nematode has been reported only on two occasions, being diagnosed in cats from Majorca Island and in northern Spain. Here, we describe a case of O. rostratus infection in a necropsied 4-year-old cat in Sicily (southern Italy). At the inspection of lungs, slender and greyish nematodes (four females and two males) were found embedded in the peri-bronchial tissues and in the bronchial walls. Parasites were morphological and molecularly identified as O. rostratus, with their 18S sequences being identical among them and showing a high homology (99%) with those available in public databases. At the histology, nematodes were encapsulated in a pseudo-cystic formation surrounded by an interstitial inflammatory process and fibrous tissue. Lung lesions were mainly represented by peri-luminal fibrosis, hyperplasia and hypertrophy of the bronchial mucosa and glands, respectively. This first record of O. rostratus infection from Italy indicates that this parasite should be included in the differential diagnosis of feline of lungworm infection.

Crenosoma brasiliense sp. n. (Nematoda: Metastrongyloidea) parasitic in lesser grison, Galictis cuja (Molina, 1782) (Carnivora, Mustelidae) from Brazil, with a key to species of Crenosoma Molin, 1861.

This study describes Crenosoma brasiliense (Nematoda, Metastrongyloidea), a new species parasitic in bronchi and bronchioles of Galictis cuja (Molina) (Carnivora, Mustelidae) from Brazil. This species differs from other 11 species of Crenosoma by having a cuticular projection at the distal end of the spicules, forming a prominent blade at the tip of the spicule, a vulval cuticular appendage with a triangular shape and prominent vulval lips. There are no previous records of species of Metastrongyloidea in G. cuja or species of Crenosoma in South America. Therefore, the new species represents the first host record and first geographical record of species of Crenosoma in South America.

Surgical extraction of an intraocular infection of Parelaphostrongylus tenuis in a horse.

CASE DESCRIPTION: A 4-year-old Hanoverian gelding was evaluated because of a mobile worm-like structure in the right eye. CLINICAL FINDINGS: Ophthalmologic examination of the right eye revealed a white, thin, coiled, mobile parasite, which was presumed to be a nematode, located in the ventral portion of the anterior chamber of the eye; there also were vitreal strands located temporally and inferiorly near the margin of the pupil. Results of ophthalmologic examination of the left eye were unremarkable. TREATMENT AND OUTCOME: The horse was treated with a neomycin-polymyxin B-dexamethasone ophthalmic solution applied topically (1 drop, q 8 h) to the right eye and penicillin V potassium (22,000 U/kg [10,000 U/lb], IV, q 6 h). The horse was anesthetized. A stab incision was made in the cornea, and a viscoelastic agent was infused around the parasite. The parasite was extracted via the incision by use of an iris hook and tying forceps. The horse had an uncomplicated recovery from the procedure and retained vision in the right eye. Gross and microscopic examination was used to identify the parasite as an adult metastrongyloid nematode consistent with a fully developed male Parelaphostrongylus tenuis. CLINICAL RELEVANCE: To the authors' knowledge, this is the first report of intraocular parelaphostrongylosis in a horse. This report provided evidence that vision could be retained after treatment for intraocular P tenuis infection in a horse.

Lymphatic dwelling filarioid nematodes in reindeer Rangifer tarandus tarandus, (cervidae) in Finland, identified as Rumenfilaria andersoni Lankester & Snider, 1982 (nematoda: Onchocercidae: Splendidofilariinae).

A filarioid nematode inhabiting the lymphatic vessels of the subserosal rumen and mesenteries associated with a high prevalence of its microfilariae in peripheral blood was observed in Finnish reindeer (Rangifer tarandus tarandus) in 2004 and 2006. Adult specimens were collected by dissecting lymphatic vessels from slaughtered animals, where some of the nematodes were seen through the wall of the dilated vessels as thin white winding threads obscuring the vessel. The morphology of adult worms and microfilaria is described based on light and scanning electron microscopy. These filariae belong to the subfamily Splendidofilariinae of the Onchocercidae and resemble Rumenfilaria andersoni, recovered from different host and localization, the ruminal veins of Alces alces in Canada. Comparison of paratypes of this species revealed only minor differences which were not sufficient to separate the filarioid parasitic in R. tarandus in Finland and we identify the nematode as R. andersoni. However, the findings suggest two different parasite populations. The finalizing of this taxonomic question in the future requires an integrated approach, in which the DNA-based and morphological identifications are consistent.

The lungworm, Halocercus brasiliensis (Nematoda: Pseudaliidae), from Guiana Dolphins Sotalia guianensis from Brazil with Pathological Findings.

The Guiana dolphin, Sotalia guianensis, is a small cetacean species found on the Atlantic coast of Central and South America from Honduras to Santa Catarina State, Brazil. The nematode Halocercus brasiliensis infects this cetacean, resulting in lung pathologies and death. The present study aimed to conduct a morphological and morphometric study of specimens of H. brasiliensis collected from S. guianensis from the coast of the state of Espírito Santo, Brazil. For this study, 7 dolphins were collected and examined for the presence of lung parasites. Nematodes were collected and processed for light microscopy, and lung fragments were processed for histological analysis. The nematodes were identified as H. brasiliensis according to their morphology and morphometry. The histopathological analysis revealed mineralization of the bronchiolar cartilage and inflammatory process. The parasitic infections by H. brasiliensis in S. guianensis can contribute to the debilitating status of these cetaceans, resulting in their stranding and accidental capture in fishing nets.

A new species of Parafilaroides (Nematoda: Filaroididae) in three species of fur seals (Carnivora: Otariidae) from the Southern Hemisphere.

Blocks of frozen lungs of 2 Australian fur seals (Arctocephalus pusillus doriferus Wood Jones, 1925), 2 New Zealand fur seals (A. forsteri [Lesson, 1828]), and 1 sub-Antarctic fur seal (A. tropicalis [Gray, 1872]) from 3 different locations (Australia, New Zealand, and South Africa, respectively) were examined and found to contain lung parasites. This represents the first thorough description and identification of a new species, Parafilaroides normani, from an eared seal (Otariidae) in the Southern Hemisphere. The new species is described, illustrated, and differentiated from the 6 recognized species in the genus by body size, spicule shape and size, vulva to anus length, and vaginal sphincter musculature.

Severe Verminous Pneumonia Caused by Natural Mixed Infection with Aelurostrongylus abstrusus and Angiostrongylus chabaudi in a European Wildcat from Western Balkan Area.

AIMS: Aelurostrongylus abstrusus (Railliet, 1898) and Angiostrongylus chabaudi (Biocca, 1957) are important cardiopulmonary metastrongyloids in felids. This case report describes, for the first time, a natural and patent mixed infection caused by A. abstrusus and A. chabaudi in a European wildcat (Felis silvestris silvestris) from Bosnia and Herzegovina. Most important, this is a rare report of fatal parasitism in wildlife, involving a severe verminous bronchopneumonia and gastrointestinal nematode and cestode infection with Toxocara cati, Taenia taeniaeformis, Aonchotheca putorii, and Ancylostoma spp. RESULTS: Emphasis is set to detailed description of granulomatous, interstitial verminous bronchopneumonia, morphological description of A. abstrusus and A. chabaudi, and molecular confirmation of diagnosis by triplex PCR. CONCLUSION: The data provided in this study contribute to the knowledge on the epizootiology and pathological effect of these neglected metastrongyloids in European wildcat.

Larval development of Aelurostrongylus abstrusus in experimentally infected Rumina decollata snails.

Aelurostrongylus abstrusus is a lungworm distributed worldwide that affects wild and domestic cats, causing bronchopneumonia of varying intensity. Snails serve as intermediate hosts. The aim of the present study was to assess the larval development of A. abstrusus in R. decollata snails and to investigate its potential as an intermediate host. For this purpose, first-stage larvae (L1) of A. abstrusus were obtained from the faeces of naturally infected cats. Doses of 500 L1/snail were given to 24 R. decollata snails, placed on the soil of the breeder chamber, and maintained under laboratory conditions. Three snails were killed at 8, 10, 12, 16, 22, 26, 45 and 55 days post-infection (dpi), and the muscular foot and visceral body were separately digested by an artificial digestion technique. The morphometric parameters of different larval stages were recorded. The mean number of larvae reaching the infective stage at the end of the study (L3) was 262 larvae/snail. The greatest development to L3 was recorded from days16 to 55 pi, during which the isolation was maximum. A. Abstrusus L3 were isolated from the viscera, but isolation from the snail foot was significantly higher. Our results showed for the first time the ability of A. Abstrusus larvae to develop in R. decollata, thus serving as a potential intermediate host.

Identification of first-stage dorsal-spined lungworm larvae of Tunisian barbary red deer: First report of Varestrongylus sagittatus and Elaphostrongylus cervi in Africa.

Barbary red deer (Cervus elaphus barbarus) is a protected rare subspecies of red deer. The study of its Protostrongylidae fauna based only on sporadic necropsy of naturally dead animals is difficult. Therefore diagnosis of lungworms rely mainly on the identification of the first stage larvae (L1). The L1 of the different species are not readily diagnosed on morphological basis since much variation is recorded within and among dorsal-spined larvae belonging to various species. The aim of this study was to identify the dorsal-spined lungworm larvae of the Barbary red deer. A discriminant function was established, using the measurements of L1 lungworms recorded from red deer in the literature, then applied to identify 220 dorsal-spined larvae extracted from 25 Tunisian Barbary red deer fresh fecal samples. Also the ITS2 region of rDNA of four pools of larvae (n = 25-60) were amplified, sequenced and analyzed. Using discriminant analysis of morphological traits, Elaphostrongylus cervi and Varestongylus sagittatus were identified. Molecular identification confirmed the presence of E. cervi which is the most prevalent species. This study represents the first identification of V. sagittatus and E. cervi in North Africa.

The first report of Aelurostrongylus falciformis (Schlegel, 1933) (Nematoda, Metastrongyloidea) in badger (Meles meles) in Poland

Aelurostrongylus falciformis belongs to the superfamily of Metastrongyloidea. This nematode occurs in European badgers and locates in lungs, in respiratory tract. Numerous species of land snails are intermediate hosts of the parasite. In 2015, parasitological necropsy of 9 badgers, shot in the Forest District Gleboki Bród in Augustowska Primeval Forest, was performed. Two examined animals were infected with nematodes A. falciformis. In the lungs of each badger two specimens of nematodes were detected (male and female). In the following article, description, morphometrical data and figures are presented. This is the first report of A. falciformis infection in badgers in Poland.

Eosinophilic encephalomyelitis in horses caused by protostrongylid parasites.

Four thoroughbred horses showing lameness, ataxia, circling, depression, recumbency, and seizures, were examined. The horses had gross, pale- to dark-red manifestations and foci in the central nervous system (CNS). Multifocal to coalescing eosinophilic necrotizing encephalomyelitis was observed histologically in the CNS along with intact or degenerated nematodes. Nematodes had polymyarian-coelomyarian musculature, a smooth thin cuticle, and intestines lined by multinucleated cells with microvilli. These traits suggested the nematodes belonged to the family Protostrongylidae, which includes Parelaphostrongylus tenuis. It was concluded that the horses were infected by nematodes, presumably Parelaphostrongylus tenuis, resulting in eosinophilic necrotizing encephalomyelitis.

Restoration of Parafilaroides (Dougherty, 1946) (Nematoda: Metastrongyloidea) with description of two new species from pinnipeds of eastern central Pacific.

The lungs of a northern elephant seal, Mirounga angustirostris (Gill, 1866), and Pacific harbor seal, Phoca vitulina richardii (Gray, 1864), from the eastern central Pacific, central California coast, were examined for parasites. This represents the first thorough description and identification of Parafilaroides species from northern elephant seals and Pacific harbor seals on the west coast of North America. From this study, 2 new species of Filaroides (Parafilaroides) are described, illustrated, and differentiated by shape of vaginal sphincter, body morphometrics, and spicule shape from the 4 existing, recognized species in the subgenus. In addition, the subgenus Parafilaroides is proposed to be reestablished to full generic status in the Filaroididae based on the presence of caudal papillae and DNA.

A new species of Skrjabingylus Petrov, 1927 (Nematoda: Metastrongyloidea) from the frontal sinuses of the hooded skunk, Mephitis macroura (Mustelidae).

Skrjabingylus santaceciliae n. sp. is described based on specimens from the frontal sinuses of a hooded skunk, Mephitis macroura, collected from the Area de Conservación Guanacaste, Costa Rica. Skrjabingylus santaceciliae n. sp. differs from the other 5 species in the genus in having pointed spicule tips that lack a rounded or lobed formation and by lacking a prominent distal projection at the tail tip. Morphometric comparisons show that S. santaceciliae n. sp. is much smaller than the only other valid species from Mephitis, Skrjabingylus chitwoodorum Hill, 1939. Likewise, morphometric comparisons also distinguish S. santaceciliae n. sp. from other described Skrjabingylus species.

Metastrongyloid infection by Aelurostrongylus abstrusus, Troglostrongylus brevior and Angiostrongylus chabaudi in a domestic cat.

The "cat lungworm", Aelurostrongylus abstrusus, affects cats worldwide. Recently, other cardio-pulmonary parasites, e.g. Troglostrongylus brevior and Angiostrongylus chabaudi, have been isolated from cats either for the first time or a long time after they were first described. This paper describes the first known mixed infection by A. abstrusus, T. brevior and A. chabaudi in a domestic cat. Biological and epidemiological implications of old and "new" metastrongyloid-caused infections in cats are discussed.

Migration and growth of Neostrongylus linearis (Marotel, 1913) Gebauer, 1932 in sheep.

In order to establish the mammalian parasitic cycle of the nematode Neostrongylus linearis 18 lambs were infected with 5000 third-stage larvae obtained from the snail Cernuella (Xeromagna) cespitum arigonis. In the subsequent days, larvae were found in the walls of the caecum and colon, in the liver and the lungs, suggesting that the bloodstream is the main migratory route. The presence of low numbers of migrating larvae in the mesenteric lymph nodes indicates that this route is probably secondary. The first still sexually undifferentiated, fourth-stage larvae were found in the lungs on the 8th day post-infection (p.i.) and the first sexually differentiated fifth-stage larvae, on the 10th day. Also described are the macroscopic changes of the parasitized organs.