1. Investigación

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 multi-locus sequence typing approach, including the ITS1/5.8S/ITS2 region (ITS), ribosomal RNA small subunit (18S) and Fe-hydrogenase gene (FeHyd). Global prevalence for 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). MLST analysis showed five different MLS types, being ITS-A/18S-VI/FeHyd-A1 and ITS-D/18S-II/Fe-C4 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. Disconrdances between samples cultured in TYM and samples subjected to PCR from oropharyngeal swabs were observed, being swab-ITS-PCR more sensitive.

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-cateninlike 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.

Mycoplasma conjunctivae is an obligate microparasite that causes Infectious Keratoconjunctivitis (IKC) in Caprinae species. IKC is a long-recognised disease, but little attention has been paid to the mechanisms of transmission of the mycoplasma and its occurrence in locations other than the eyes. In this study, the presence of M. conjunctivae is assessed in the eyes, external ear canals (EEC), nasal cavity, and vagina of host species as well as in potential vectors, which may be involved in the transmission and persistence of infection within the host. M. conjunctivae was detected by qPCR in 7.2 % (CI 95% 4.7-11.0) of the ear swabs and 9.5 % (CI 95% 6.4-13.9) of the nasal swabs from Pyrenean chamois, Iberian ibex, domestic sheep and mouflon without statistical differences between species. Mycoplasma detection in nasal swabs was mostly associated with ocular infection (95.6%), but this was not the case for EEC (52.6%). Among the eye-positive ruminants, 27.3% were positive in ear swabs and 64.7% in nasal swabs, and the threshold cycle values of the qPCR were correlated only between eye and nasal swabs (p<0.01; r2=0.56). M. conjunctivae was detected in 1.7% - 7.1 % of Musca spp. captured during an IKC outbreak in Iberian ibex and in one out of three endemic sheep flocks. The results indicate that the transmission of M. conjunctivae may occur by direct contact with eye or nasal secretions and/or indirectly through flies. The M. conjunctivae DNA detection in EEC suggests that it can colonise the auditory tract, but the significance for its persistence within the host should be further assessed.

Studying the causes of parasite geographic distribution is relevant to understand ecological and evolutionary processes that affect host populations as well as for species conservation. Temperature is one of the most important environmental variables affecting parasite distribution, as raising temperatures positively affect development, reproduction, and rate of transmission of both endo- and ectoparasites. In this context, it is generally accepted that, in mountains, parasite abundance decreases with elevation. However, empirical evidence on this topic is limited. In the present study, we analyzed the elevational variation of hemoparasites and ectoparasites of a lizard, Psammodromus algirus, along a 2,200-m elevational gradient in Sierra Nevada (SE Spain). As predicted, ectoparasite (mites, ticks, mosquitoes, and sandflies) abundance decreased with elevation. However, hemoparasite prevalence and intensity in the lizard augmented with altitude, showing a pattern contrary to their vectors (mites). We suggest that tolerance to hemoparasites may increase with elevation as a consequence of lizards at high altitudes taking advantage of increased body condition and food availability, and reduced oxidative stress. Moreover, lizards could have been selected for higher resistance against hemoparasites at lowlands (where higher rates of replication are expected), thus reducing hemoparasite prevalence and load. Our findings imply that, in a scenario of climate warming, populations of lizards at high elevation may face increased abundance of ectoparasites, accompanied with strong negative effects.