CHARACTERIZATION OF LESIONS INDUCED BY SPIROCERCA VULPIS (SPIRURIDAE: SPIROCERCIDAE) IN RED FOXES (VULPES VULPES).

Spirocerca lupi infection in dogs (Canis domesticus) is associated with esophageal lesions that may evolve to a neoplastic stage in the form of esophageal sarcoma. In the red fox (Vulpes vulpes) infected with the closely related Spirocerca vulpis, similar lesions may occur in the stomach, but neoplastic forms have not been reported. We characterize Spirocerca vulpis-induced lesions in the fox, using pathology and immunohistochemical (IHC) techniques. Seventy-one out of 163 Spirocerca vulpis-positive red foxes were selected and subjected to histopathological study. Lesions were classified as patchy or diffuse. Ten patchy and 10 diffuse lesion samples were studied using three IHC markers (CD68, CD3, and CD79α for macrophages, T lymphocytes, and B lymphocytes, respectively) and H&E stain for neutrophils and eosinophils. Intensity of necrosis, hemorrhages, and the presence of collagen was also analyzed. Of the S. vulpis-positive red foxes, 96.9% had S. vulpis nodules localized in the gastric area (wall and/or omentum), and 3.1% had nodules in the small intestine. All the samples had a moderate to severe lymphoplasmacytic infiltrate. Mild eosinophil infiltration was observed in both types of lesions, while neutrophil infiltration was significatively higher in the patchy than in the diffuse lesions. Fibrosis with mature collagen fibers was also predominant in the patchy lesions along with the presence of T lymphocytes and macrophages. Both the patchy and diffuse patterns had very few B lymphocytes. These findings suggest that the diffuse form is an earlier stage of the lesion, which eventually evolves into patchy forms. Neoplastic forms were not seen. Although more studies are necessary, this study describes the lesions, characterizes the inflammatory infiltrates, and establishes a possible evolution of the different pathological forms of S. vulpis infection in the red fox.

First detection of Gongylonema species in Geotrupes mutator in Europe.

The detection of three Gongylonema sp. infective larvae in two specimens of the dung beetle Geotrupes mutator (Marsham, 1802) from western Spain is reported here for the first time in Europe. Scanning electron microscopy confirmed that the analyzed specimens belong to the genus Gongylonema, but it was not possible to determine the species identity by the lack of morphological information in the literature and because many of the phenotypic characteristics had not yet fully developed at this juvenile stage. Nevertheless, a phylogenetic analysis using amplified cox1 nucleotide sequences has revealed that the studied larvae could be clearly discriminated (< 89% identity) from all the other Gongylonema cox1 sequences available in public genetic databases. While our results are limited by the scarcity of genetic information available for this genus, the possibility that the analyzed specimens might correspond to a new species should not be ruled out, and more studies are needed. The results provided in this report indicate that G. mutator is involved in the transmission cycle of Gongylonema sp. to vertebrates in Europe.

Primeras detecciones de Aedes albopictus (mosquito tigre) en la región de Extremadura, oeste de España / First detections of Aedes albopictus (tiger mosquito) in the region of Extremadura, west of Spain

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Culex pipiens as a potential vector for transmission of Dirofilaria immitis and other unclassified Filarioidea in Southwest Spain.

Dirofilaria immitis is one of the most frequently detected mosquito-transmitted zoonotic filarioid nematode in mammals in Europe, being canine dirofilariosis a major animal health problem, endemic in the Mediterranean area. This study, focused on Southwest Spain, in order to bring new insights into (i) the epidemiology of Dirofilaria spp., (ii) the species of Culicid vectors possibly involved in their transmission and (iii) the genetic variability of those potential vectors. A total of 881 adult female mosquitoes from 11 different species, were captured during 2012-2013, and detection of filarioid DNA was attempted by PCR using specific primers (ITS-2 and COI), followed by DNA sequencing. In a single Culex pipiens specimen D. immitis DNA was detected both in the head-thorax and abdomen sections. Filarioid nematode DNA was also detected in eight additional Cx. pipiens specimens also in both the thorax and the abdomen, but analysis of sequence data did not allow unambiguous assignment of any of the obtained sequences to a previously defined species. All Cx. pipiens with filarioid DNA were individually analysed by CQ11 to discriminate between pipiens, molestus, and hybrid forms. Besides, rDNA ITS-2 sequence analysis revealed the presence of haplotype H1 and H2 of Cx. pipiens. To our knowledge this study revealed, for the first time in Spain, the occurrence of likely mature infection of D. immitis in Cx. pipiens, as well as with other yet uncharacterized nematodes, supporting its role as a potential vector of these filarids.

First molecular detection of Leishmania tarentolae-like DNA in Sergentomyia minuta in Spain.

Phlebotomine sand flies (Diptera, Psychodidae) are vectors of multiple Leishmania species, among which Leishmania infantum stands out as a being frequently pathogenic to humans and dogs in Mediterranean countries. In this study, Sergentomyia minuta sand flies were collected using CDC miniature light traps in different 431 biotopes from Southwest Spain. A total of 114 females were tested for the presence of Leishmania DNA by targeting ITS-1 and cyt-B sequences by PCR. Leishmania DNA was detected in one S. minuta. Characterization of the obtained DNA sequences by phylogenetic analyses revealed close relatedness with Leishmania tarentolae Wenyon, 1921 as well as with both human and canine pathogenic strains of Asian origin (China), previously described as Leishmania sp. To our knowledge, this is the first report of phlebotomine sand flies naturally infected with L. tarentolae-like in Spain. The possible infection of sand flies with novel Leishmania species should be taken into consideration in epidemiological studies of vector species in areas where leishmaniosis is endemic.

First report of Thelazia callipaeda in wildlife from Spain.

We describe the first cases of infection by the nematode, Thelazia callipaeda (Spirurida, Thelaziidae) haplotype 1 in two red foxes (Vulpes vulpes) in Spain and discuss the potential role of red foxes as a reservoir for T. callipaeda.

Supplemental feeding drives endoparasite infection in wild boar in Western Spain.

Wildlife population management is thought to destabilize existing host-parasite equilibriums in opposing directions, that is, it may increase parasite success or host resilience once infection takes place. This process is of special importance for species such as the wild boar (Sus scrofa) that are managed for game purposes throughout much of Europe. However, little is known about how this practices influcences either gastrointestinal or pulmonary parasitism in the wild boar. Twelve hunting estates were chosen in order to study the relationship of management measures (feeder density, wild boar abundance, the ratio of wild boar per feeder and the percentage of sclerophyllous vegetation) and host factors (age and sex) with gastrointestinal and pulmonary parasite aggregation, richness, infection probability and intensity of infection. Parasitological analyses from 300 wild boar gastrointestinal and 269 respiratory tracts were performed for this purpose. A set of general linear models with combinations of the explanatory variables was built and the model with the smallest Akaike Information Criterion was selected as the best. The feeder density increased gastrointestinal parasite traits (richness, infection probability and intensity of infection), probably due to the contamination of feeding sites with infective parasite forms. Pulmonary parasite traits, on the other hand, were only influenced by host sex and age class, and parasite aggregation was as expected for a wild population. Managers should be aware of the consequences on parasitism when implementing supplemental feeding in hunting estates.

Epidemiologic study of lung parasites (Metastrongylus spp.) in wild boar (Sus scrofa) in southwestern Spain.

We analyzed 927 wild boars (Sus scrofa) in southwestern Spain during the hunting seasons of 2004/2005 to 2008/2009. Respiratory tracts were examined for lung nematodes (Metastrongylus spp.). The prevalence of Metastrongylus spp. was 41.1%. The most frequently isolated species were Metastrongylus apri (71.4%), Metastrongylus pudendotectus (28.0%), and Metastrongylus salmi (0.6%). Prevalence and infection intensity were greater in young animals (<1 yr old) than in older animals. There were no significant differences in prevalence between sexes. Prevalence and intensity of infection were higher in areas of high altitude and high rainfall.

Congenital toxoplasmosis in wild boar (Sus scrofa) and identification of the Toxoplasma gondii types involved.

Congenital toxoplasmosis has been little described in wild animals. We report a case of vertical transmission in wild boar (Sus scrofa). Necropsy and histopathologic examination of a pregnant female and her three fetuses revealed all to have lesions compatible with acute toxoplasmosis. Nested polymerase chain reaction B1 gene detected Toxoplasma gondii in maternal (heart and diaphragm) and fetal (central nervous system, retina, optic nerve, heart, lung, tongue, and diaphragm) samples. The mother had a mixed infection of T. gondii types I and III. One fetus with type III infection developed no malformations, but the others-one with type I infection and one infected by types I and III-showed bilateral ocular agenesis, prognathism, and agenesis of the nasal cartilage. These results suggest the pathogenicity of the various T. gondii types may differ in wild boars.