Unusual parasitoses in urban dogs: Urinary capillariosis and linguatulosis, cases report.

Parasitic diseases caused by uncommonly diagnosed parasites may pose a threat to companion animals' health in urban environments where they are least expected. The pentastomid Linguatula serrata (tongue worm) and the capillarid Pearsonema plica (syn. Capillaria plica) are parasites with an indirect life cycle, infecting both domestic and wild carnivores. The present report describes two cases: the first one of urinary capillariosis and the other of linguatulosis, in two dogs living in the urban environment of Athens, Greece. In the case of capillariosis, the dog never lived out of the city, so it was presumably infected in the urban environment. On the contrary, in the case of linguatulosis, the dog was adopted at a young age from a rural environment but remained asymptomatic for several months while living in the city. Both dogs had mild symptoms, compatible with these infections. Urinary capillariosis and linguatulosis are uncommon in owned, pet dogs, living in cities due to epizootiological characteristics, i.e. need for wildlife reservoir for P. plica and consumption of raw infected viscera for L. serrata. Different factors contribute to the fact that such infections may occur in scenarios where they are least expected. Recent studies show a progressive worldwide increase in the number of uncommon parasitoses in pet animals, that in some cases, such as linguatulosis, are of zoonotic relevance. Regular parasitological examinations and preventive antiparasitic schemes are necessary in order to treat and prevent infections in pet animals and safeguard the health of both animals and humans under the concept of One Health.

Ticks, Fleas, and Harboured Pathogens from Dogs and Cats in Cyprus.

Ticks and fleas are blood-sucking ectoparasites that cause irritation and anaemia to their hosts and act as vectors of pathogens (vector-borne pathogens, VBPs) of relevance for animal and human health. In the present study, tick and flea species in dogs and cats from Cyprus were recorded and VBPs were detected in the collected specimens. Ectoparasites were collected from 220 animals (161 dogs and 59 cats), and a questionnaire including demographic, clinical, and other information was filled out for each animal. The ectoparasites were morphologically identified and the detection of VBPs was performed by PCR-coupled sequencing. Rhipicephalus sanguineus sensu lato was found on 108 dogs and 13 cats, and Ixodes gibbosus on 2 dogs. Ctenocephalides felis was the predominant flea species (on 62 dogs and 45 cats), while one dog and one cat were infested by Ctenocephalides canis and Echidnophaga gallinacea, respectively. The VBPs in ticks were Anaplasma platys, Rickettsia massiliae, Rickettsia conorii, Rickettsia felis, Hepatozoon felis and Hepatozoon canis, while Rickettsia felis, Rickettsia sp., Bartonella koehlerae, Bartonella clarridgeiae, and Bartonella henselae were recorded in fleas. Statistical analysis (chi-square test and multiple univariate generalized linear model) showed that animals up to 6 months of age were less likely to be infested with ticks than older animals, but more likely to be infested with fleas. Ticks were more prevalent in sheltered than in owned animals, while the odds ratio of flea presence was higher in owned animals than those living in shelters. The present study is the first investigation on the occurrence of ticks and fleas in dogs and cats from Cyprus, showing the presence of different VBPs in these important ectoparasites. The results point out the importance of systematic ectoparasite control in dogs and cats.

Investigation of the Food-Transmitted Parasites Trichinella spp. and Alaria spp. in Wild Boars in Greece by Classical and Molecular Methods and Development of a Novel Real-Time PCR for Alaria spp. Detection.

Foodborne parasitic diseases represent a major threat to public health. Trichinellosis, caused by the nematode parasite Trichinella spp., is one of the most important foodborne diseases, while alariosis, caused by the trematode parasite Alaria spp., is less common in humans, and rare cases have been reported only in the USA and Canada. Both parasites can infect humans via the consumption of raw or undercooked wild boar meat. In order to investigate the prevalence of these parasites in wild boar meat in Greece, samples from the diaphragm pillars and the region of the mandibular angle from 128 wild boars, hunted in Greece, were collected. The samples were examined by classical parasitological (compression, artificial digestion, and Alaria spp. migration) and by molecular (real-time PCR) methods. For Trichinella spp. an existent real-time PCR detecting all species likely to be present in Greece was applied, while for Alaria spp. a real-time PCR was developed, employing an LNA TaqMan probe targeting the large subunit ribosomal RNA gene. All examined wild boar samples from Greece resulted negative for Trichinella and Alaria species, indicating a low prevalence of infection in the examined population. The novel real-time PCR for Alaria spp. has 81.5% amplification efficiency and is able to detect 0.12 larvae per 50 g of tissue and could be utilized as a complementary to AMT diagnostic tool in surveillance.

Endoparasites of European Wildcats (Felis silvestris) in Greece.

The European wildcat (Felis silvestris) is the only wild felid living in Greece. Wildcat populations are declining due to anthropogenic and phenological unfavourable conditions, and parasites may have an additional negative impact. In the present study, the occurrence of endoparasites in wildcats in Greece and the potential threats posed to wildcats, domestic animals, and humans in the study areas has been investigated. In a six-year period, 23 road-killed wildcats and 62 wildcat faecal samples were collected from different areas of the country. Necropsy for the detection of endoparasites and standard parasitological examinations of faecal samples were performed. Parasites were morphologically identified and, in selected cases, molecularly analysed. All necropsied wildcats (100%) were infected by three to 10 different parasite taxa, with the most prevalent being Taenia taeniaeformis (73.9%), Toxocara cati (60.9%), Angiostrongylus chabaudi (56.5%), Ancylostoma tubaeforme (39.1%), Cylicospirura spp. (34.8%), Troglostrongylus brevior (34.8%), and Capillaria aerophila (33.8%). Of the 62 faecal samples examined, 53 (85.5%) were positive for one or more parasite elements (larvae, eggs, or oocysts). The most frequent were T. cati (45.2%), A. chabaudi (29%), C. aerophila (24.2%), and Ancylostomatidae (17.7%). This is the first survey on endoparasites affecting wildcats in Greece. Some of the parasites here found are frequent in domestic and wild felids, while others, i.e., Oslerus rostratus and Cylicospirura petrowi, were described for the first time in the European wildcat. Most of them have a significant pathogenic potential, causing severe to hazardous diseases to infected felids and some, under specific circumstances, can also threaten human health.

Cat Respiratory Nematodes: Current Knowledge, Novel Data and Warranted Studies on Clinical Features, Treatment and Control.

The nematodes Aelurostrongylus abstrusus, Troglostrongylus brevior and Capillaria aerophila are the most important parasites inhabiting the airways of cats. They are receiving growing attention from academia, pharmaceutical companies and veterinarians, and are now considered a primary cause of respiratory diseases in feline clinical practice and parasitology. In the past few years, several studies have been conducted in both natural and experimental settings to increase knowledge, provide new insights and fill gaps on respiratory parasitoses of cats. Awareness and knowledge of clinical scenarios towards appropriate and timely diagnosis and prompt and efficacious treatment options have become a priority to investigate. At the same time, chemopreventative approaches have been evaluated to assess the geographical spreading of these parasites and the rise in the number of clinical cases in cat populations of different countries. Given the intense accumulation of novel data, this review presents and discusses the state of the art and the latest updates on the clinical features, treatment, and control of major respiratory parasitoses of cats. Moreover, food for thought is also provided with the aim of spurring on new studies in the near future.

First record of Hepatozoon spp. in domestic cats in Greece.

Feline hepatozoonosis is an emerging disease of domestic and wild felids though there is limited knowledge of this infection, e.g. regarding geographical distribution and parasite species involved. The present study evaluated microscopically and molecularly the occurrence of Hepatozoon spp. in domestic cats from insular (Crete, Mykonos and Skopelos) and continental (prefectures of Attica and Thessaloniki) Greece. Out of 282 cats examined, 72 (25.5 %) scored positive by PCR for Hepatozoon spp. and of them, 9 (12.5 %) showed gamonts on the blood smear microscopic examination. Sequences obtained from 35 of the amplicons proved the presence of two haplotypes of Hepatozoon felis. One, herein called H1 (34/35 amplicons) resulted 100 % identical with H. felis from Italy and isolates from other continents, and ∼98 % similar with a H. felis isolate causing severe clinical signs in Austria. The haplotype H2, found in a cat in Skopelos, had ∼94 % identity with H1, with H. felis isolates from Italy, Israel, Spain, a ∼92 % identity with the isolate from Austria, and ∼94-98 % with isolates from South Africa. These are the first records of H. felis in cat populations from Greece and indicate that the infection may be present at high prevalences in different regions of the country. Furthermore, the results of the molecular and phylogenetic analysis support a recent hypothesis indicating the existence of a species-complex classification for H. felis. Further studies aiming at elucidating the genetic make-up of Hepatozoon populations and possible variations in terms of geographic distribution and clinical relevance are necessary. The importance of a continuous epizootiological monitoring is crucial for the establishment of preventative and control measures protecting the health of cats living in or travelling to enzootic areas.

Leishmania infantum Seroprevalence in Cats From Touristic Areas of Italy and Greece.

Leishmaniosis by Leishmania infantum is a major zoonotic Vector-Borne Disease (VBD) in terms of geographic distribution, pathogenicity and zoonotic potential. While dogs are the main reservoir of L. infantum, the infection in cats is poorly understood although increasingly reported from enzootic and non-enzootic areas. The Mediterranean basin is a key area for leishmaniosis and includes touristic spots that require continuous surveillance for VBDs in consideration of the growing tendency of tourists to travel with their pets. This study evaluated L. infantum seroprevalence in cats living in selected touristic localities of Italy and Greece. A total of 269 cat serum samples from three Sites i.e., 76, 40, and 153 from Adriatic Coast of Abruzzo, Italy (Site A), Giglio Island, Tuscany, Italy (Site B), and Mykonos Island, Greece (Site C), respectively, were included in the survey. Sera samples were subjected to an indirect immunofluorescence antibody assay for the detection of anti-L. infantum specific IgG. Associations between possible risk factors and seropositivity to L. infantum were statistically evaluated. Antibodies against L. infantum were detected in eight out of 269 (3.0%) cats tested i.e., 4/76 (5.3%), 1/40 (2.5%), and 3/153 (2.0%), from sites A, B, and C, respectively. A statistical association between anti-L. infantum antibodies and cohabitation with dogs was shown. This study indicates that feline populations living in the examined Italian and Greek touristic areas are exposed to L. infantum and that they may contribute to the circulation of L. infantum, enhancing the risk of infection for dogs and humans.

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.

Clinical investigations and treatment outcome in a European wildcat (Felis silvestris silvestris) infected by cardio-pulmonary nematodes.

Recently, feline cardio-pulmonary nematodes have attracted high scientific interest, as they are increasingly reported from various areas. Most of these parasites have similar transmission patterns and/or host reservoirs, thus they may affect domestic and wild felids living in sympatry. In the present study, a case of multiple cardio-pulmonary parasitism in co-infection with other parasites in a European wildcat is presented. The animal, found exhausted, was hospitalised for recovery and parasitological, haematological, clinical and imaging examinations were performed. The parasitological examinations revealed 4 cardio-pulmonary nematodes, i.e. Aelurostrongylus abstrusus, Troglostrongylus brevior, Eucoleus aerophilus, Angiostrongylus chabaudi, 3 intestinal parasites, i.e. Toxocara cati, ancylostomatids, Cystoisospora felis, 2 haemoparasites, i.e. Hepatozoon felis and elements morphologically compatible with small Babesia/Cytauxzoon spp., and Ixodes ricinus and Haemaphysalis erinacei ticks. Treatment with a spot-on formulation containing imidacloprid 10% and moxidectin 1% (Advocate® spot-on solution for cats, Bayer) was decided and follow-up faecal examinations were performed until the release of the animal. By the end of the hospitalisation, all metazoan endoparasites were no longer detectable in faecal examinations, with the exception of a low number of A. abstrusus larvae. Thus, the animal was released after a second treatment with the same product. This is the first description of an apparently successful treatment of multiple cardio-respiratory parasitosis in a naturally infected wildcat showing compatible clinical signs. The evidence that Advocate® may be effective against A. chabaudi could be useful for treating infected, hospitalised, wildcats and it is promising in the case A. chabaudi infection will spread to domestic cats in a near future.

Filarial infections in dogs in Cyprus, an apparently heartworm free island.

The current study investigated for the first time the occurrence of filarial infections in dogs in Cyprus. Blood samples of dogs from five districts of Cyprus (Lefkosia, Lemessos, Larnaka, Pafos and Ammochostos) were examined by the modified Knott's method and by serology, and the morphological classification of microfilariae was confirmed by molecular methods. A total of 200 dogs, 153 living in shelters and 47 owned dogs, were included in the study. Acanthocheilonema reconditum microfilariae were found in 9 (4.5%) samples and one (0.5%) sample was seropositive for D. immitis. No statistical significance was observed between microfilaraemic samples and the evaluated variables apart from the district (p = .005). The present study showed that dogs in Cyprus can be infected with blood circulating microfilariae and for the first time A. reconditum was reported as autochthonous infection in dogs in the country. No microfilariae of Dirofilaria spp. were detected. However, veterinarians should remain vigilant regarding Dirofilaria infections and should consider preventive protection to the animals, at least in case of travel in enzootic areas.

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.