Efficacy of Bravecto® Plus spot-on solution for cats (280 mg/ml fluralaner and 14 mg/ml moxidectin) in the prevention of feline Aelurostrongylus abstrusus infection evaluated in a multi-diagnostic approach.

BACKGROUND: Aelurostrongylus abstrusus is one of the most important respiratory nematodes of felines. Infections may lead to respiratory clinical signs with varying severity or even death, emphasizing the need for preventive treatment of cats with outdoor access to circumvent patent infections. METHODS: Therefore, the preventive efficacy of a spot-on formulation of 280 mg/ml fluralaner and 14 mg/ml moxidectin (Bravecto® Plus spot-on solution for cats, MSD) against A. abstrusus was evaluated in a negative controlled, randomized and partially blinded efficacy study with 28 purpose-bred cats in a non-terminal design. In three different treatment regimes, the minimum recommended dose of 40 mg fluralaner and 2.0 mg moxidectin/kg bodyweight (BW) was administered once at 12, 8 or 4 weeks (study group G1, G2 and G3, respectively) prior to experimental infection with 300 third-stage A. abstrusus larvae, while G4 served as placebo-treated control. RESULTS: From 30 to 46 days post infection (dpi; SD 114 to 130), faeces were sampled to monitor first-stage larvae (L1) excretion for efficacy determination. Secondary efficacy criteria, including respiratory parameters, serological antibody levels and computed tomography (CT) findings, were assessed once before enrolment (SD -7 to -1) and before infection (SD 75 to 83). After infection, CT evaluation was performed once at 47-50 dpi (SD 131 to 134), and respiratory parameters and antibody levels were regularly assessed twice or once a week, respectively (1 up to 78 dpi, SD 85 up to 162). All animals in the control group excreted L1 by 33-37 dpi and remained positive throughout the study period from 41 to 46 dpi (SD 125 to 130). In the treatment groups, only one animal each of G1 and G2 excreted L1 at two consecutive days, and four cats of G1, two of G2 and three of G3 were positive on single occasions. While the geometric mean (GM) of the maximum number of excreted L1 per 5 g of faeces was 7380.89 in the control group (G4), GMs were significantly lower in the treatment groups with 1.63 in G1, 1.37 in G2 and 0.79 in G3. Thus, based on GMs, the reduction in excreted L1 exceeded 99.9% in all three treatment groups. Based on CT severity scores, all lungs of the animals of the control group showed severe pulmonary changes post infection, whereas lungs of the cats of the treatment groups were either unaltered (4 animals), mildly (11 animals), or moderately altered (5 animals). Moreover, seroconversion was observed in all cats of the control group, but not in those of the treatment groups. CONCLUSIONS: The combination of diagnostic methods used in this non-terminal study yielded coherent and reliable results. A single administration of Bravecto® Plus spot-on solution for cats was well tolerated and effective in the prevention of aelurostrongylosis for at least 12 weeks.

Infectivity of gastropod-shed third-stage larvae of Angiostrongylus vasorum and Crenosoma vulpis to dogs.

BACKGROUND: Metastrongyloid parasites Angiostrongylus vasorum and Crenosoma vulpis infect wild and domestic canids and are important pathogens in dogs. Recent studies indicate that gastropod intermediate hosts infected with various metastrongyloids spontaneously shed infective third-stage larvae (L3) into the environment via feces and mucus under laboratory conditions. Shed L3 retain motility up to 120 days, but whether they retain infectivity was unknown. METHODS: To assess the infectivity of shed L3, the heart/lungs of six red foxes (Vulpes vulpes) were obtained from trappers in Newfoundland, Canada. Lungs were examined for first-stage larvae (L1) by the Baermann technique. A high number of viable A. vasorum L1 and a low number of C. vulpis L1 were recovered from one fox; these were used to infect naïve laboratory-raised Limax maximus. L3 recovered from slugs by artificial digestion were fed to two naïve purpose-bred research beagles (100 L3/dog). L1 shed by these two dogs was used to infect 546 L. maximus (2000-10,000 L1/slug). L3 shedding was induced by anesthetizing slugs in soda water and transferring them into warm (45 °C) tap water for at least 8 h. Shed L3 recovered from slugs were aliquoted on romaine lettuce in six-well tissue culture plates (80-500 L3/well) and stored at 16 °C/75% relative humidity. Four naïve research beagles were then exposed to 100 L3/dog from larvae stored for 0, 2, 4, or 8 weeks, respectively, after shedding. RESULTS: All four dogs began shedding C. vulpis L1 by 26-36 days post-infection (PI). All four dogs began shedding A. vasorum L1 by 50 days PI. CONCLUSIONS: L3 infectivity for the definitive host was retained in both metastrongyloids, indicating the potential for natural infection in dogs through exposure from environmental contamination. As an additional exposure route, eating or licking plant or other material(s) contaminated with metastrongyloid L3 could dramatically increase the number of dogs at risk of infection from these parasites.

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.

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.

First Report of a Troglostrongylus brevior Case in a Domestic Cat in Turkey

Parasitic bronchopneumonia plays an important role, especially in the respiratory diseases of cats. Troglostrongylus species have been reported as lung parasites of wild cats; however, in recent years, they have also been found in domestic cats, especially from the Mediterranean and Balkan countries. In this report, Troglostrongylus brevior was found during the examination of lung samples of a tabby cat who was euthanized at the age of 2 years. To the best of our knowledge, this is the first report of Troglostrongylus brevior in Turkey.

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.

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.

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.

Clinical crenosomosis in a black bear (Ursus americanus).

An orphaned black bear (Ursus americanus) cub, estimated to be 9 months-of-age was presented to a wildlife rehabilitation facility in December of 2016. The cub was afebrile, under-weight (6.8 kg) and had a cough condition. Centrifugal sugar fecal flotation examination failed to detect any gastrointestinal helminth or protozoan parasites, but revealed the presence of first-stage nematode larvae (L1). Large numbers of L1 (>8000 L1/g) identified as Crenosoma sp. based on morphology were recovered using the Baermann technique. Three species (Crenosoma petrowi, Crenosoma potos, Crenosoma vulpis) have been reported from black bears. Based on larval length measurements (range = 253-277 µm; mean = 263 µm; n = 8), the L1 were tentatively identified as C. petrowi. Further molecular characterization using Polymerase Chain Reaction (PCR) and DNA sequencing of the small subunit (SSU) RNA gene and two regions of the large subunit (LSU) rRNA gene did not match any submissions in GenBank, but were most similar to Crenosoma mephiditis. There is a paucity of molecular data for members of the genus Crenosoma, with only information for Crenosoma vulpis (red fox), C. mephitidis (skunks), Crenosoma striatum (hedgehog) and Crenosoma sp. (red panda) in GenBank. Molecular analysis eliminates C. vulpis as a possibility in this case but due to the lack of submissions in GenBank, the identification of the L1 as C. petrowi based on length measurements could not be confirmed. Receiving in total, three separate courses of treatment with fenbendazole (50 mg/kg, oral, once a day for 3 days), fecal larval shedding ceased and clinical signs resolved. The black bear cub was released into the wild in June 2017. This is the first report of clinical chronic respiratory disease due to Crenosoma sp. infection in a black bear.

First report of Aelurostrongylus abstrusus in St. Kitts.

A 4-month-old intact male domestic shorthair kitten living in St Kitts, West Indies presented with respiratory distress, cachexia, and mucopurulent nasal discharge. Thoracic radiographs revealed a diaphragmatic hernia. The diaphragmatic hernia as well as subpleural pulmonary nodules suspicious for verminous pneumonia were identified during a postmortem examination. Histology showed multifocal to coalescing pyogranulomatous and eosinophilic pneumonia centered on larvae and morulated eggs. The lesion and nematode morphology were consistent with Aelurostrongylus abstrusus. Although Aelurostrongylus abstrusus has been reported worldwide, this is the first report of a metastrongyloid lungworm in cats in St. Kitts and for the West Indies. This case report should increase the awareness of A. abstrusus pneumonia in cats from St. Kitts and other locations in the eastern Caribbean.

Why Use the Mini-FLOTAC to Detect Metastrongyloid Larvae in Dogs and Cats?

PURPOSE: Canine and feline cardiopulmonary disease is an emergent condition in several European countries, particularly in non-endemic regions, although it is often underestimated because of the limitations of the common diagnostic techniques. The aim of this study was to evaluate the performance of Mini-FLOTAC to detect cardiopulmonary larvae from dogs and cats compared to reference techniques such as Baermann and FLOTAC. METHODS: A total of 20 fresh faecal samples were collected from ten dogs and ten cats naturally infected with Angiostrongylus vasorum, and Aelurostrongylus abstrusus and Troglostrongylus spp., respectively. Three replicates were performed for each technique. FS3 (zinc sulfate, specific gravity = 1.200) was used as flotation solution for Mini-FLOTAC and FLOTAC. The results were expressed as the arithmetic mean of LPG (larvae per gram of faeces). RESULTS: The mean LPG calculated by means of Mini-FLOTAC was significantly higher (P < 0.05) than that obtained through the Baermann technique for A. vasorum, A. abstrusus and Troglostrongylus spp., whereas it was significantly higher (P < 0.05) than that obtained through the FLOTAC technique only for A. vasorum and A. abstrusus. CONCLUSIONS: Mini-FLOTAC can be considered a valid alternative for the detection of metastrongyloid larvae of dogs and cats, overcoming the limitation of time required by the Baermann test.

Effect of different temperatures on survival and development of Aelurostrongylus abstrusus (Railliet, 1898) larvae.

Aim of the study was to get in-depth information on Aelurostrongylus abstrusus first-stage larvae (L1s) survival at different temperatures and to assess the capability of these larvae to develop into the third infective stage (L3s). Faeces of a naturally infected cat were split into two aliquots: the first was divided in subsamples assigned to four groups (F1-F4); from the second aliquot, L1s were extracted by Baermann technique, suspended in water and divided into four groups (W1-W4). Groups were stored at different temperatures (F1/W1 -20 ± 1°C; F2/W2 +4 ± 1°C; F3/W3 +14 ± 1°C; and F4/W4 +28 ± 1°C) and L1s vitality assessed every seven days. The capability of L1s stored in water to develop into L3s in snails was evaluated at the beginning and every 21 days. The L1s of W2 and F2 groups remained viable for a longer period (231 and 56 days, respectively) compared to those of other groups. The capability of L1s to moult into L3s in snails showed a decreasing trend; the group W2's L1s maintained the capability to moult into L3s for the longest time (day 189) compared to the other groups. The time of survival of A. abstrusus L1s is influenced by temperature. However, the species seems to be more resistant to temperature variations than other feline lungworms, and this may explain its wider distribution across Europe.

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.

Efficacy of a moxidectin/imidacloprid spot-on formulation (Advocate®) for the treatment of Troglostrongylus brevior in naturally infected cats in a field study in Greece.

BACKGROUND: Troglostrongylus brevior is a lungworm of wild felids that recently has been recognized as agent of severe respiratory disease in domestic cats in Mediterranean and Balkan countries. Nevertheless, the information on treatment options for feline troglostrongylosis is still poor. The aim of this pilot field trial was to evaluate the efficacy of the spot-on formulation containing 1% w/v moxidectin and 10% w/v imidacloprid (Advocate® spot-on solution for cats, Bayer Animal Health GmbH) in the treatment of T. brevior infection in naturally infected cats in Greece. METHODS: The trial was a negative control, multicentre, clinical efficacy study conducted according to the standards of Good Scientific Practice (GSP). Sixteen cats in two study sites, naturally infected with T. brevior, were allocated to an untreated control group (G1, n = 8) or a treatment group (G2, n = 8), according to a randomization list. Animals assigned to G2 were treated with Advocate® for cats on days 0 and 28 at the recommended dose rate and animals assigned to G1 received a rescue treatment with the same product on days 56 and 84. Efficacy was assessed on days 28 and 56 in G2 and on days 84 and 112 in G1 by faecal larval counts. The primary efficacy criterion was the absence of T. brevior first-stage larvae (L1) following treatment. Other efficacy parameters were the quantitative comparison of L1 presence before (baseline) and after one or two treatments in both groups. RESULTS: All G2 cats were negative for T. brevior L1 at the first post-treatment evaluation (100% efficacy) while G1 cats were persistently shedding L1. The difference of the mean number of L1 per gram between G2 and G1 was statistically significant (P < 0.001). All G1 cats were negative (100% efficacy) for T. brevior L1 at the first post-rescue-treatment evaluation. Therefore, treatment efficacy at study completion was 100% in both groups in terms of stopping the L1 shedding in the faeces of the animals. No adverse effects were observed during the study. CONCLUSIONS: These results indicate that Advocate® spot-on solution for cats represents an option for treating cats naturally infected with T. brevior.

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.

Crenosoma vulpis in red foxes (Vulpes vulpes) in Northern Italy.

Crenosoma vulpis, the fox lungworm, is a nematode parasite of wild and domestic canids belonging to the super-family Metastrongyloidea. A survey of infection was carried out examining 88 red foxes (Vulpes vulpes) obtained during the regular hunting season (2014-2015) from the Emilia-Romagna region of Italy. Carcasses were stored frozen (- 21 °C) prior to necropsy. Lungs were examined for the presence of adult worms by dissection of the trachea, bronchi, and bronchioles, and then the lung tissue was examined for first-stage larvae (L1) by the Baermann method. No adult stages were detected, but L1, identified based on morphology as Crenosoma vulpis, were recovered from 28.4% (25/88) of the fox lungs. No significant differences in infection were found based on sex or geographical distribution. A brief review on C. vulpis report in red foxes in Italy and other European countries was also carried out.

The invasive giant African snail Lissachatina fulica as natural intermediate host of Aelurostrongylus abstrusus, Angiostrongylus vasorum, Troglostrongylus brevior, and Crenosoma vulpis in Colombia.

BACKGROUND: Several metastrongyloid lungworms are unreported pathogens in Colombia. Angiostrongylus vasorum and Crenosoma vulpis target the cardiopulmonary system of domestic and wild canids. Aelurostrongylus abstrusus and Troglostrongylus brevior infect felids and considering that six wild felid species exist in Colombia, knowledge of feline lungworm infections is important for their conservation. The zoonotic metastrongyloids Angiostrongylus costaricensis and Angiostrongylus cantonensis can cause severe gastrointestinal and neurological diseases. Angiostrongylus costaricensis has been reported in Colombia, while Ang. cantonensis is present in neighbouring countries. Research on the epidemiology of metastrongyloids in Colombia and South America more broadly requires evaluating the role that gastropods play as intermediate hosts in their life cycles. This study assessed the prevalence of metastrongyloid larvae in populations of the invasive giant African snail, Lissachatina fulica, in Colombia. METHODOLOGY/PRINCIPAL FINDINGS: A total of 609 Lissachantina fulica were collected from 6 Colombian municipalities. The snails were then cryo-euthanized, artificially digested and the sediments examined microscopically for the presence of metastrongyloid larvae. Based on morphological characteristics 53.3% (56/107) of the snails from Puerto Leguízamo (Department of Putumayo) were infected with Ael. abstrusus larvae, 8.4% (9/107) with Ang. vasorum larvae, 6.5% (7/107) with T. brevior larvae and 5.6% (6/107) with C. vulpis larvae, being the region with highest prevalences of the four species. Snails from Andes (Department of Antioquia) and Tulúa (Department of Valle del Cauca) were positive for Ang. vasorum larvae with a prevalence of 4.6 (11/238) and 6.3% (4/64), respectively. Species identifications were confirmed by PCR and sequencing. CONCLUSIONS/SIGNIFICANCE: This epidemiological survey reports for first time the presence of Ael. abstrusus, T. brevior, C. vulpis and Ang. vasorum in L. fulica in a number of regions of Colombia.