Efficacy of milbemycin oxime/afoxolaner chewable tablets (NEXGARD SPECTRA®) against Capillaria aerophila and Capillaria boehmi in naturally infected dogs.

BACKGROUND: Capillaria aerophila and Capillaria boehmi parasitize the respiratory system of wild and domestic carnivores. Capillaria aerophila inhabits the trachea and bronchi of dogs and cats, while C. boehmi affects the nasal cavities and sinuses of dogs. In dogs the infection may be subclinical or characterized by varying respiratory distress. METHODS: The present study evaluated the efficacy of an oral formulation containing milbemycin oxime and afoxolaner (NEXGARD SPECTRA®) in dogs naturally infected with C. aerophila and/or C. boehmi from three enzootic areas of Italy. Dogs were enrolled pending fecal examination and molecular confirmation of respiratory capillarioses. Dogs were allocated in two groups: Group 1 (G1, 25 dogs), treated with a negative control product with no anthelmintic activity (afoxolaner, NEXGARD®), and Group 2 (G2, 26 dogs), treated with NEXGARD SPECTRA®. At the day of treatment administration (Day 0), all dogs were clinically examined. Dogs were again subjected to clinical and fecal examinations at Days 28 (± 4) and 56 (± 2). The primary criterion for treatment efficacy was the reduction of fecal Capillaria egg counts in G2 compared with G1. The regression of/recovery from baseline clinical signs was considered as a further efficacy criterion. RESULTS: Percentage reduction of fecal Capillaria egg counts in the NEXGARD SPECTRA® group compared to the control group was > 97% on Day 28 and 100% on Day 56, respectively (p < 0.05 for both time points). Twelve of the 13 dogs in the NEXGARD SPECTRA® group with respiratory signs prior to treatment were free of clinical signs at the end of the study. Conversely, the six control group dogs with respiratory signs prior to treatment remained symptomatic. CONCLUSIONS: Results of the present study showed that NEXGARD SPECTRA® was safe and highly efficacious in the reduction of C. aerophila and C. boehmi eggs after one treatment with a complete reduction of the egg output after the second administration associated with a recovery from respiratory signs.

Molecular characterization of Calodium hepaticum in grey dwarf hamster (Cricetulus migratorius).

Calodium hepaticum (Bancroft, 1893) Moravec, 1982 (Syn. Capillaria hepatica) is a zoonotic nematode that causes hepatic capillariosis, an uncommon zoonotic infection. The nematode is globally distributed and parasitizes the liver of mammals, mainly Muroidea. Cricetulus migratorius Pallas, 1773 (Cricetinae) was rarely reported as a host for C. hepaticum. In Turkey, C. hepaticum was recorded in three rodent species; Rattus rattus, R. norvegicus, and Apodemus flavicollis. In this study, C. migratorius (grey dwarf hamster) has been identified as a new host species for C. hepaticum in Turkey. The parasite was identified by morphological, histological, and molecular methods and the phylogenetic relationships of C. hepaticum collected from different hosts were revealed. This is the first molecular characterization of C. hepaticum from a grey dwarf hamster.

Capillaria Ova and Diagnosis of Trichuris trichiura Infection in Humans by Kato-Katz Smear, Liberia.

We examined human stool samples from Liberia for soil-transmitted helminth ova by Kato-Katz smear and by quantitative PCR. Twenty-five samples were positive for Trichuris trichiura by smear but negative by quantitative PCR. Reexamination of samples showed that they contained Capillaria eggs that resemble T. trichiura in Kato-Katz smears.

Calodium hepaticum (Nematoda: Capillariidae) in wild rodent populations from Argentina.

Calodium hepaticum (Nematoda; Capillariidae) is a parasitic nematode of mammals with a cosmopolitan distribution. Adults of this nematode can infect the liver of many mammalian species, including humans, but the main hosts are members of the superfamily Muroidea. Among these, Rattus spp. have the highest apparent prevalences reported worldwide. There are only two reports of C. hepaticum infecting Sigmodontinae species (Muroidea: Cricetidae). In this survey, we examined the occurrence of C. hepaticum in two assemblages of Sigmodontinae rodents from Argentina (Santa Fe and Entre Ríos provinces). The diagnosis was made by morphological features, histopathological exam, and molecular characterization of 18S ribosomal RNA gene region. Here, we show that C. hepaticum is a common parasite of Sigmodontinae. We report the infection in six species (all new hosts): Akodon azarae, Calomys callidus, Calomys venustus, Oligoryzomys flavescens, Oligoryzomys nigripes, and Oxymycterus rufus. This is the first report of C. hepaticum in Sigmodontinae rodents from Argentina and the second record in this subfamily for South America. It is also the first confirmation of C. hepaticum infection in Sigmodontinae by molecular diagnosis. The genetic findings and the prevalences observed, together with the existing information on C. hepaticum, lead us to propose that Rattus spp. brought C. hepaticum to the New World exposing Sigmodontinae rodents, which are frequently infected by this parasite. The high prevalence of infection in A. azarae (41.2%) suggests that this host may be playing an important role in C. hepaticum dynamics in the New World.

Zoonotic intestinal parasites and vector-borne pathogens in Italian shelter and kennel dogs.

This study investigated the presence of zoonotic parasites and vector-borne pathogens in dogs housed in kennels and shelters from four sites of Italy. A total of 150 adoptable dogs was examined with different microscopic, serological and molecular methods. Overall 129 dogs (86%) were positive for one or more parasites and/or pathogens transmitted by ectoparasites. Forty-eight (32%) were positive for one infection, while 81 (54%) for more than one pathogen. The most common zoonotic helminths recorded were hookworms, roundworms and Capillaria aerophila, followed by mosquito-borne Dirofilaria spp. and Dipylidium caninum. One hundred and thirteen (77.9%), 6 (4.1%) and 2 (1.4%) dogs were positive for Rickettsia spp., Leishmania infantum and Anaplasma spp., respectively. The results show that dogs living in rescue facilities from the studied areas may be infected by many zoonotic internal parasites and vector-borne pathogens, and that control measures should be implemented.

High prevalence of Eucoleus boehmi (syn. Capillaria boehmi) in foxes from western Austria.

Eucoleus boehmi (syn. Capillaria boehmi) is a canine trichuroid nematode affecting the upper respiratory airways (i.e., nasal cavity and paranasal sinuses) of dogs, foxes, and wolves. In the past few years, reports in dogs and wild canids have increased from across Europe, but data on its occurrence and distribution in Austria is scanty. A total of 47 red foxes (Vulpes vulpes) from the two westernmost provinces (Tyrol and Vorarlberg) of Austria were therefore examined for the presence of E. boehmi at necropsy. Eggs and adult nematodes were identified morphologically and molecularly (cox1) as E. boehmi. These nematodes were found in 26 (78.8 %) and 13 (92.9 %) foxes from Tyrol and Vorarlberg, respectively, with an overall prevalence of 83.0 % (39/47). The prevalence rate of infection recorded in this study is among the highest in Europe. These results suggest that foxes may represent an important source of infection for dogs and other canids, but further studies are needed to elucidate the transmission dynamics.

Morphological and molecular genetic characterization of three Capillaria spp. (Capillaria anatis, Capillaria pudendotecta, and Capillaria madseni) and Baruscapillaria obsignata (Nematoda: Trichuridae: Capillariinae) in avians.

Morphological and genetic analyses were performed on four avian species of the subfamily Capillariinae (Nematoda: Trichuridae), i.e., Capillaria anatis from chickens (Gallus gallus domesticus) in Japan and the Philippines, Baruscapillaria obsignata from chickens and captive swans (Cygnus olor and Cygnus atratus) in Japan, Capillaria pudendotecta from captive swans in Japan, and Capillaria madseni from carrion and jungle crows (Corvus corone and Corvus macrorhynchos) in Japan. Although morphometric variations of male and female worms from different hosts and/or localities made the species identification difficult, the 18S ribosomal RNA gene (rDNA) sequences clarified their taxonomic position and phylogenetic relationships. Species of the same genus clustered robustly into a single clade in the phylogenetic tree based on the 18S rDNA, demonstrating to the extent possible the validity of the latest classification of the subfamily following Moravec's rearrangement in 1982. Male worms of C. pudendotecta are described here for the first time.

Molecular detection of Capillaria philippinensis: An emerging zoonosis in Egypt.

Human infection with Capillaria philippinensis is accidental; however, it may end fatally if not diagnosed and treated in the proper time. The first case was detected in the Philippines in 1963, but later reported in other countries around the world, including Egypt. In this report, molecular diagnosis using a specific nested PCR for detection of C. philippinensis in faeces is described based on the amplification of small ribosomal subunit. The test showed sensitivity and specificity, as it detected all the positive cases and gave no cross-reaction with human DNA and DNA of other tested parasites. This method can be very useful not only for improvement of diagnosis, but also to understand the different environmental routes of transmission by detection of C. philippinensis DNA-stages in the possible fish intermediate hosts and reservoir animal host, helping to improve strategies for surveillance and prevention of human disease.

Molecular phylogeny of Pseudocapillaroides xenopi (Moravec et Cosgrov 1982) and development of a quantitative PCR assay for its detection in aquarium sediment.

We used high-fidelity PCR to amplify a portion of the small ribosomal subunit (18S rRNA) of Pseudocapillaroides xenopi, a nematode that parasitizes the skin of Xenopus laevis. The 1113-bp amplicon was cloned, sequenced, and aligned with sequences from 22 other nematodes in the order Trichocephalida; Caenorhabditis elegans was used as the outgroup. Maximum-likelihood and Bayesian inference phylogenetic analyses clustered P. xenopi in a clade containing only members of the genus Capillaria. Our analyses support the following taxonomic relationships: 1) members of the family Trichuridae form a clade distinct from those in the family Trichocephalida; 2) members of the genera Trichuris and Capillaria form 2 distinct clades within the family Trichuridae; and 3) the genus Trichuris includes 2 distinct clades, one representing parasites that infect herbivores and the other representing parasites that infect omnivores and carnivores. Using 18S rRNA sequence unique to P. xenopi, we developed a Taq Man quantitative PCR assay to detect this P. xenopi sequence in total DNA isolated from aquarium sediment. The assay's lower limit of detection is 3 copies of target sequence in a reaction. The specificity of our assay was validated by using negative control DNA from 9 other pathogens of Xenopus. Our quantitative PCR assay detected P. xenopi DNA in the sediment of 2 of 12 aquaria from the source institution of the specimen used to develop the assay; these aquaria had been treated with ivermectin 6 mo previously.

Molecular detection of Capillaria aerophila, an agent of canine and feline pulmonary capillariosis.

Capillaria aerophila, a trichuroid nematode causing pulmonary infections in wild and domestic carnivores, is occasionally and potentially poorly recognized in infections of humans due to clinicopathological mimicry and a lack of accurate, robust laboratory diagnostics. The present work evaluated the efficiency of a DNA-based assay amplifying a partial cytochrome c oxidase subunit 1 (cox1) gene of C. aerophila in the diagnosis of lung capillariosis. Fecal samples from 34 dogs and 10 cats positive at parasitological examination for C. aerophila and other endoparasites (i.e., other lungworms, whipworms, roundworms, hookworms, tapeworms, and/or coccidia) and from 44 animals negative for C. aerophila but positive for other endoparasites were molecularly examined. Of the 44 samples positive for C. aerophila at copromicroscopy, 43 scored positive (i.e., 33/34 dogs and 10/10 cats) in seminested PCR, resulting in a sensitivity of 97 to 100%. Samples that were copromicroscopy negative for C. aerophila although positive for other endoparasites never produced a PCR product or nonspecific amplicons. The specific PCR amplification of C. aerophila (i.e., specificity of 100%) was confirmed by a nucleotide sequence analysis of the cox1 amplicons. The potential implications of the molecular diagnosis of lung capillariosis are discussed.

Mitochondrial DNA polymorphism within and among species of Capillaria sensu lato from Australian marsupials and rodents.

The nucleotide variation in a mitochondrial DNA (mtDNA) fragment within and among species of Capillaria sensu lato from Australian marsupials and rodents was analyzed using a mutation scanning/sequencing approach. The fragment of the cytochrome c oxidase subunit I (COI) was amplified by PCR from parasite DNA, and analysed by single-strand conformation polymorphism (SSCP) and sequencing. There was no significant variation in SSCP profiles within a morphospecies from a particular host species, but significant variation existed among morphospecies originating from different host species. The same morphospecies was found to occur in 1-3 tissue habitats within one host individual or within different individuals of a particular species of host from the same or different geographical areas, and morphospecies appeared to be relatively host specific at the generic level. The results indicated that the species of Capillaria sensu lato examined, although highly variable in their host and tissue specificity, may exhibit the greatest degree of specificity at the level of host genus.