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Toxocara spp. is one of the most common zoonotic geohelminths in the world. Its infections are associated with the accidental ingestion of contaminated soil and affecting, especially children. In this study, feces, and soil samples from 14 public parks in the city of Valencia were analyzed. The Telemann method and a modified version of a sieving technique were used to process feces and soil, respectively. None of the fecal samples and 10.9% of soil samples from five parks (35.7%) tested positive for the presence of Toxocara eggs. The most contaminated areas were the canine sanitary parks (30.8% of the samples), followed by socialization areas for dogs (9.7%); no positive samples were found at children's playgrounds. Our results suggest that most pets in Valencia are periodically dewormed, although additional preventive measures should be applied, since the risk of infection exists probably due to the presence of stray dogs and feral cats.

Avian trichomonosis is a parasitic disease that affects wild birds. The objective of this work was to determine the importance of avian trichomonosis in Bonelli's eagles to improve conservation measures in this population. One hundred and eighty-eight birds were studied: 181 chicks, two juveniles, one subadult and four adults. The birds were externally examined and gross lesions at the oropharynx registered. Samples from the oropharyngeal cavity were obtained for Trichomonas spp. detection by culture and PCR, and positive samples were subjected to a multilocus sequence typing approach, including the ITS1/5.8S/ITS2 region (ITS), ribosomal RNA small subunit (18S) and Fe-hydrogenase gene (FeHyd). Global prevalence of T. gallinae infection was 37.8% in total, 45.5% in nestlings. Thirty-three percent of the birds developed lesions that ranged from mild ( n  = 41) to moderate ( n  = 14) or severe ( n  = 7). Multilocus sequence typing analysis showed five different MLS types, ITS-A/18S-VI/FeHyd-A1 and ITS-D/18S-II/Fe-C4 being the most frequent. An association between ITS-A/18S-VI/FeHyd-A1 and moderate or severe lesions was observed, but birds with type ITS-A/18S-VI/FeHyd-A2 also developed lesions. On the contrary, birds with MLS type ITS-D/18S-II/FeHyd-C4 displayed only a low proportion of mild lesions. Chicks raised in nests were at higher risk for T. gallinae infection and development of lesions than chicks raised in captivity. Discordances between samples cultured in TYM and samples subjected to PCR from oropharyngeal swabs were observed, swab-ITS-PCR being more sensitive. RESEARCH HIGHLIGHTS 45.5% of Bonelli's eagles in the nest carried T. gallinae and 39.4% showed lesions.PCR from oral swabs showed higher sensitivity than culture in TYM for detection of T. gallinae .MLS types ITS-A/18S-VI/Fe-A1 (and A2) are a risk factor for the development of lesions.

Oropharyngeal avian trichomonosis is mainly caused by Trichomonas gallinae, a protozoan parasite that affects the upper digestive tract of birds. Lesions of the disease are characterized by severe inflammation which may result in fatality by starvation. Two genotypes of T. gallinae were found to be widely distributed in different bird species all over the world. Differences in the host distribution and association with lesions of both genotypes have been reported. However, so far no distinct virulence factors of this parasite have been described and studies might suffer from possible co-infections of different genotypes. Therefore, in this paper, we analyzed the virulence capacity of seven clones of the parasite, established by micromanipulation, representing the two most frequent genotypes. Clones of both genotypes caused the maximum score of virulence at day 3 post-inoculation in LMH cells, although significant higher cytopathogenic score was found in ITS-OBT-Tg-1 genotype clones at days 1 and 2, as compared to clones with ITS-OBT-Tg-2. By using one representative clone of each genotype, a comparative proteomic analysis of the membrane proteins enriched fraction has been carried out by a label free approach (Data available via ProteomeXchange: PXD013115). The analysis resulted in 302 proteins of varying abundance. In the clone with the highest initial virulence, proteins related to cell adhesion, such as an immuno-dominant variable surface antigen, a GP63-like protein, an armadillo/beta-catenin-like repeat protein were found more abundant. Additionally, Ras superfamily proteins and calmodulins were more abundant, which might be related to an increased activity in the cytoskeleton re-organization. On the contrary, in the clone with the lowest initial virulence, larger numbers of the identified proteins were related to the carbohydrate metabolism. The results of the present work deliver substantial differences between both clones that could be related to feeding processes and morphological changes, similarly to the closely related pathogen Trichomonas vaginalis., Competing Interests: The authors have declared that no competing interests exist.

A juvenile Cinereous Vulture ( Aegypius monachus) fledgling was found disorientated on the roof of a building in Madrid City, Spain, in October 2016. A veterinary examination revealed multiple plaques distributed throughout the oropharyngeal cavity. Lesions were located under the tongue and at the choanal slit, hard palate, and esophagus opening and ranged from 2 to 7 mm, coalescing in areas up to 2 cm, with a yellowish color of the surface. Motile trichomonad trophozoites were detected in fresh wet mount smears from the lesions. Sequence analysis of the internal transcribed spacer (ITS)1/5.8S/ITS2 and small subunit ribosomal RNA confirmed that Trichomonas gypaetinii was the etiologic agent. Microbiologic cultures did not reveal any pathogenic bacteria or fungi. The animal recovered successfully after treatment with metronidazole and trimethoprim-sulfamethoxazole and was later released in a suitable habitat. Avian trichomonosis lesions caused by T. gypaetinii have not been reported.

Many obligate and facultative avian scavengers are increasingly dependent on food provided in supplementary feeding stations (SFS), which are managed for the conservation of these species. Deliberate feeding can influence disease-related host demography and population dynamics through physiological changes and density-dependent parasite acquisition and transmission, but information on this threat to avian scavengers is scarce. Due to their effects on host aggregation and density, we hypothesised that the predictability and concentration of food in SFS can exacerbate parasite infection. This hypothesis was tested by comparing the prevalence, richness, abundance and mixed infection of endoparasites (coccidia and helminths) in red kites Milvus milvus foraging on livestock carcasses (mostly of pigs and poultry) in overcrowded and confined conditions at SFS, relative to those foraging alone or in small groups on wild prey unevenly randomly distributed within large areas during winter, mostly wild rabbits (Oryctolagus cuniculus). No clear differences were found between areas with and without SFS in the prevalence and abundance of oocyst of Eimeria. This coccidian genus appears to include parasites of the prey rather than the raptors, thus representing parasite transport or pseudo-parasitism rather than actual parasitism in the kites. A higher prevalence and richness of helminths, as well as mixed infections with several phyla, was found in kites exploiting SFS than in those feeding on wild prey in the area without SFS. The unsanitary conditions derived from the stack of livestock carcasses and the contamination of carrion with the faeces of multiple scavenger hosts can increase the accumulation and persistence of helminths eggs and intermediate hosts. The regular use and frequent confinement of large numbers of red kites at SFS can promote the spread of parasites to a large proportion of the European breeding population distributed across Spain during the winter. We encourage that carcasses of free roaming livestock can be left in the countryside, as well as the conservation management of wildlife exploited as food by red kites (especially wild rabbits), to attempt avoiding overcrowded and confined conditions at SFS. Further research is required to assess the impact of deliberate feeding on the spread of parasites and other disease agents in the threatened species SFS are intended to favour., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Oropharyngeal trichomonad isolates of wild birds from Spain were studied. A total of 1688 samples (1214 of predator birds and 474 of prey species) from wildlife recovery centres and scientific bird-ringing campaigns were analysed from 2011 to 2013. The overall infection prevalence was 20.3% (11.4% in predator birds and 43.3% in prey species). Pathognomonic lesions were present in 26% of the infected birds (57.3% in predator birds and 4.9% in prey species). The most commonly parasitized species were the goshawk (Accipiter gentilis, 74.5%) and the rock pigeon (Columba livia, 79.4%). Host species in which the parasite has not been previously analysed by polymerase chain reaction and sequencing in Spain are also reported: Columba palumbus, Streptopelia turtur, Pica pica, A. gentilis, Accipiter nisus, Asio otus, Bubo bubo, Buteo buteo, Circus aeruginosus, Circus cyaneus, Falco naumanni, Falco peregrinus, Neophron percnopterus, Otus scops, Pernis apivorus and Strix aluco. Sequence analysis of the ITS1/5.8S/ITS2 region revealed five different genotypes and also some mixed infections. A relationship between genotype and host species was observed, but only two genotypes (ITS-OBT-Tg-1and ITS-OBT-Tg-2) were widely distributed. Genotype ITS-OBT-Tg-1 was most frequently found in predator birds and statistically associated with pathognomonic lesions. Non-strict ornithophagous species were at higher risk to develop disease than ornithophagous ones. Genotypes ITS-OBT-Tcl-1 and ITS-OBT-Tcl-2 are new reports, and ITS-OBT-Tvl-5 is reported for the first time in Spain. They showed higher genetic homology to Trichomonas canistomae and Trichomonas vaginalis than to Trichomonas gallinae, indicating the possibility of new species within this genus.

Avian trichomonosis is a world-wide parasitic disease caused by the protozoan Trichomonas gallinae. Although several degrees of pathogenicity have been described on the basis of the clinical signs in birds, there are few reports concerning the genetic characterization of the parasite and its relationship with pathogenicity. The parasite usually appears apathogenic but is occasionally responsible for outbreaks of the disease in avian populations, particularly affecting nestlings of ornithophagous raptors. For 3 years, cultures of oropharingeal samples from 612 wild and domestic pigeons (Columba livia) and 102 birds of prey from 15 different species were made in an attempt to determine the prevalence of T. gallinae in the Valencian Community (eastern Spain). To establish the genotype of the isolates, 5.8S rRNA and the surrounding internal transcribed spacer regions were amplified by polymerase chain reaction and were sequenced. After restriction map analysis, sequencing and polymerase chain reaction-restriction fragment length polymorphism using HaeIII showed two genotypes (A and B) in isolates from both groups of birds, although genotype prevalence differed in each group-genotype A being more prevalent in columbiforms and genotype B in raptors. In addition, genotype B was present in every bird that displayed macroscopic lesions.