Publications by authors named "Eva Frontera"

14 Publications

  • Page 1 of 1

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

J Wildl Dis 2021 Nov 17. Epub 2021 Nov 17.

Universidad de Extremadura, Facultad de Veterinaria, Departamento de Sanidad Animal, Parasitología, Avda. Universidad s/n, 10003, Cáceres, Spain.

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.
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http://dx.doi.org/10.7589/JWD-D-20-00162DOI Listing
November 2021

First detection of species in in Europe.

J Nematol 2021 21;53. Epub 2021 May 21.

Parasitology Area, Department of Animal Health, Veterinary Faculty, University of Extremadura, Cáceres, Spain.

The detection of three sp. infective larvae in two specimens of the dung beetle (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 , 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 nucleotide sequences has revealed that the studied larvae could be clearly discriminated (< 89% identity) from all the other 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 is involved in the transmission cycle of sp. to vertebrates in Europe.
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http://dx.doi.org/10.21307/jofnem-2021-050DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8138951PMC
May 2021

West Nile and Usutu virus infections in wild birds admitted to rehabilitation centres in Extremadura, western Spain, 2017-2019.

Vet Microbiol 2021 Apr 24;255:109020. Epub 2021 Feb 24.

Animal Health Department, Veterinary Faculty, University of Extremadura (UEx), Cáceres, Spain. Electronic address:

West Nile virus (WNV) is an emerging flavivirus transmitted generally by mosquitoes of Culex genus. It is maintained in an enzootic life cycle where birds act as reservoir hosts. Humans and horses are also susceptible to infection, and occasionally, they suffer from neurological complications. However, they do not transmit the virus to other vectors, behaving as dead-end hosts. Sporadic WNV outbreaks observed in horses and wild birds from Extremadura (western Spain) during 2016 and 2017 seasons prompted to carry out this survey in wild birds, focused on specimens coming from two wildlife rehabilitation centres. Between October 2017 and December 2019, samples from 391 wild birds, belonging to 56 different species were collected and analysed in search of evidence of WNV infection. The analysis of serum samples for WNV-specific antibodies by ELISA, whose specificity was subsequently confirmed by virus-neutralisation test (VNT) showed positive results in 18.23 % birds belonging to 18 different species. Pelecaniformes (33.33 %), Accipitriformes (25.77 %) and Strigiformes (22.92 %) orders had the higher seroprevalences. Remarkably, WNV-specific antibodies were found in a black stork for the first time in Europe. Analysis by real time RT-PCR in symptomatic birds confirmed the presence of WNV lineage 1 RNA in griffon vulture and little owls. Specificity analysis of ELISA positive and doubtful sera was performed by differential VNT titration against WNV and two other cross-reacting avian flaviviruses found in Spain: Usutu virus (USUV) and Bagaza virus (BAGV). Only four samples showed USUV-specific antibodies (1.04 %) corresponding to three species: Eurasian eagle-owl, griffon vulture and great bustard (first detection in Europe) whereas no samples were found reactive to BAGV. Differential VNT yielded undetermined flavivirus result in 16 samples (4.17 %). This is the first study carried out on wild birds from Extremadura (western Spain). It highlights the widespread circulation of WNV in the region and its co-circulation with USUV.
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http://dx.doi.org/10.1016/j.vetmic.2021.109020DOI Listing
April 2021

Serological evidence of co-circulation of West Nile and Usutu viruses in equids from western Spain.

Transbound Emerg Dis 2021 May 5;68(3):1432-1444. Epub 2020 Sep 5.

Animal Health Department, Veterinary Faculty, University of Extremadura (UEx), Cáceres, Spain.

West Nile virus (WNV) is a mosquito-borne emerging virus in Europe with capacity to cause neurological complications such as encephalitis or meningoencephalitis in humans, birds or equids. In Spain, WNV is actively circulating in mosquitoes, birds and horses in different regions, but never has been deeply studied in Extremadura. Therefore, the aim of this study was to evaluate the seroprevalence of WNV in equids of those areas and to analyse the risk factors associated with exposure to the virus. A total of 199 out of 725 equids presented antibodies against WNV by competition ELISA (27.45%), while 22 were doubtful (3.03%). Anti-WNV IgM antibodies were detected in 16 equids (2.21%), and 3 animals were doubtful (0.41%). All ELISA-reactive positive/doubtful sera (N = 226) were further tested by micro-virus neutralization test (VNT), and a total of 143 horses were confirmed as positive for WNV, obtaining a seroprevalence of 19.72% in equids of western Spain. In addition, specific antibodies against USUV were confirmed in 11 equids. In 24 equids, a specific flavivirus species (detected by ELISA test) could not be determined. The generalized linear mixed-effects models showed that the significant risk factors associated with individual WNV infection in equids were the age (adults) and hair coat colour (light), whereas in USUV infections, it was the breed (pure). Data demonstrated that WNV and USUV are circulating in regions of western Spain. Given the high WNV seroprevalence found in equids from the studied areas, it is important to improve the surveillance programmes of public health to detect undiagnosed human cases and to establish a vaccination programme in equid herds in these regions.
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http://dx.doi.org/10.1111/tbed.13810DOI Listing
May 2021

Histological and Immunohistochemical Study of Wounds in Sheep Skin in Maggot Therapy by Using Protophormia terraenovae (Diptera: Calliphoridae) Larvae.

J Med Entomol 2020 02;57(2):369-376

Animal Medicine Department, Pathological Anatomy Unit, University of Extremadura. Avenida de la Universidad SN. Cáceres, España.

A study was performed to evaluate the implication of Protophormia terraenovae larvae as a surgical therapy for wounded skin. Three groups of sheep (n = 25) were considered based on larval doses. Groups 1 and 2 were artificially infested with low and high concentrations of L1 stage P. terraenovae, respectively, and group 0 served as a control. Skin biopsies were taken at 4 and 14 d postinfestation (D.P.If). A histopathological study was carried out to evaluate the lesions with a score, numbers of eosinophils and mast cells, and an immunohistochemical analysis of CD3, CD79α, and CD68 as T lymphocytes, B lymphosytes, and macrophages, respectively. The results indicated that higher larval doses led to faster regeneration by 14 D.P.If. Furthermore, the higher larval doses showed a high number of the CD68 marker and eosinophils and a low number of CD3 and CD79α markers and mast cells. In addition, the number of mast cells, T lymphocytes, and macrophage markers increased when the lesion progressed; however, a low number of immunolabeled CD79α cells and eosinophils were observed. The results indicate a possible positive effect of larvae in the healing of certain wounds.
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http://dx.doi.org/10.1093/jme/tjz185DOI Listing
February 2020

Geographic dispersal and genetic diversity of tick-borne phleboviruses (Phenuiviridae, Phlebovirus) as revealed by the analysis of L segment sequences.

Ticks Tick Borne Dis 2019 06 3;10(4):942-948. Epub 2019 May 3.

Unidade de Microbiologia Médica, (IHMT/UNL, and GHTM), Oeiras, Portugal. Electronic address:

The large diversity of new tick-borne phleboviruses, and the negative impacts of the virulent viruses on human/animal health have led to a growing interest in their analysis. In this report, new insights are brought out into the diversity of putative phleboviruses circulating in Portugal (both the continental territory and the islands of São Miguel, in the Azores, and Madeira), as well as in the Spanish western regions of Extremadura and Castilla and León. Phlebovirus sequences were frequently detected (L-segment) from both questing and feeding ticks, but especially in Rhipicephalus sanguineus sensu lato (s.l.) specimens. These sequences were detected in adult ticks, as well as nymphs and eggs, supporting the hypothesis of viral maintenance by vertical transmission. Though multiple genetic groups could be identified in phylogenetic trees (AnLuc, KarMa, RiPar virus 1, and Spanish group 1 and 2), all the sequences from Portugal and Spain shared common ancestry with other viral sequence obtained from samples collected over a large geographic coverage. Spatiotemporal analysis placed Middle-East as the geographic origin of the most recent common ancestor (MRCA) of all phleboviruses analysed in the present study. More recent viral transitions might include migrations from Spain to continental Portugal, and from there to the Portuguese Islands. Our findings suggest that the time of the MRCA of phleboviruses was dated around 225 years ago [95% HPD: 124-387 year before the last sampling date].
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http://dx.doi.org/10.1016/j.ttbdis.2019.05.001DOI Listing
June 2019

[First detections of Aedes albopictus (tiger mosquito) in the region of Extremadura, west of Spain].

Gac Sanit 2019 May - Jun;33(3):299-300. Epub 2018 Dec 24.

Unidad de Parasitología y Enfermedades Parasitarias, Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, España.

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http://dx.doi.org/10.1016/j.gaceta.2018.11.003DOI Listing
December 2019

The mosquito fauna of the western region of Spain with emphasis on ecological factors and the characterization of Culex pipiens forms.

J Vector Ecol 2017 06;42(1):136-147

Global Health and Tropical Medicine, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Unidade de Parasitologia Médica, Rua da Junqueira 100, 1349-008 Lisboa, Portugal.

This study updates the diversity, distribution, and seasonal trends of mosquitoes in a western region of Spain, assesses ecological determinants of Culex pipiens s.l., and determines form composition of Cx. pipiens s.s.

Populations: A total of 1,495 mosquitoes of 16 species was collected during 2012-2013, of which Cx. pipiens s.l. and Cx. theileri were the most abundant. Five new records for An. maculipennis s.s., Orthopodomyia pulcripalpis, Aedes (Ochlerotatus) punctor, Cx. europaeus, and Cx. modestus were found for this region. Cx. pipiens density varied across weather and habitat patterns, correlating positively with high temperatures and with a preference for urbanized areas and rural areas within a proximity of ovine farms. Moreover, molecular identification by CQ11FL was performed in 467 Cx. pipiens s.s., detecting both pipiens (66%) and molestus (8.4%) forms coexisting in different habitats (urban, peri-urban and rural) aboveground with a high degree of hybridization (25.7%). The abundance of Cx. pipiens in urban areas and farms, with the presence of hybrids, may increase their capacity to act as bridge vectors for the transmission of arboviral infections. These data will be helpful for further implementation of entomological programs focused on risk assessment for arboviruses or other mosquito-borne pathogens.
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http://dx.doi.org/10.1111/jvec.12248DOI Listing
June 2017

Comparative morphological and molecular analysis confirms the presence of the West Nile virus mosquito vector, Culex univittatus, in the Iberian Peninsula.

Parasit Vectors 2016 11 25;9(1):601. Epub 2016 Nov 25.

Global Health and Tropical Medicine, GHTM, UEI Medical Parasitology, Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNL, Lisbon, Portugal.

Background: Culex univittatus and Culex perexiguus mosquitoes (Diptera: Culicidae) are competent arbovirus vectors, but with unclear morphological differentiation. In Europe, and in the Iberian Peninsula in particular, the presence of either or both species is controversial. However, in order to conduct adequate surveillance for arboviruses in this region, it is crucial to clarify whether Cx. univittatus is present or not, as well as to critically assess existing differentiation tools. This study aimed to clarify this situation, by morphological and molecular phylogenetic comparison of Iberian specimens deemed as Cx. univittatus, with others of South African origin, i.e. from the type-locality region.

Methods: Thus, morphological characteristics useful to distinguish both species, such as midfemur pale line, hindfemur R ratio, seta g R ratio, seta f shape, length of ventral arm of phalosome and number of setae on IX tergal abdominal segment, were observed. A phylogenetic analysis based on cox1 mtDNA, of which there were no sequences from Cx. univittatus yet available in the GenBank database, was performed.

Results: This analysis showed that Iberian and South African specimens are morphologically similar, except for the length of the ventral arm of the phalosome, which was higher in the Iberian specimens. Although the Iberian specimens could not be accurately identified using BOLD Systems, phylogenetic analysis still grouped these closer to South African Cx. univittatus, than to Cx. perexiguus from Turkey and Pakistan, despite the observed segregation of both taxa as two individual monophyletic clusters with shared common ancestry.

Conclusions: This survey demonstrates that the West Nile virus vector Cx. univittatus is present in the Iberian Peninsula.
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http://dx.doi.org/10.1186/s13071-016-1877-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5123335PMC
November 2016

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

Vet Parasitol 2016 Jun 27;223:173-80. Epub 2016 Apr 27.

Parasitology and Parasitic Diseases, Animal Health Department, Veterinary Faculty, University of Extremadura, Spain. Electronic address:

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.
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http://dx.doi.org/10.1016/j.vetpar.2016.04.030DOI Listing
June 2016

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

Parasitol Res 2016 Mar 22;115(3):1339-44. Epub 2015 Dec 22.

Parasitology and Parasitic Diseases, Animal Health Department, Veterinary Faculty, University of Extremadura, Cáceres, 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.
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http://dx.doi.org/10.1007/s00436-015-4887-zDOI Listing
March 2016

Prevalence and genotype identification of Toxoplasma gondii in wild animals from southwestern Spain.

J Wildl Dis 2015 Jan;51(1):233-8

1 Parasitology Section, Animal Health Department, Veterinary Faculty, University of Extremadura, Avd. de la Universidad s/n, 10071 Cáceres, Spain.

We used PCR to detect Toxoplasma gondii in the principal game species in southwestern Spain. We detected T. gondii in 32.2% of animals tested. Prevalences varied from 14.7% in wild boar (Sus scrofa) to 51.2% in red fox (Vulpes vulpes). The most prevalent genotype was type II (50.0%), followed by type III (20.6%) and type I (5.9%). Mixed infections (11.8%) were detected in wild boar (types I+III) and red fox (types II+III). Polymorphic strains (11.8%) were detected in several species. The high prevalence and the genetic variability shown could have implications for infection of farm animals and humans.
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http://dx.doi.org/10.7589/2013-09-233DOI Listing
January 2015

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

J Wildl Dis 2013 Oct;49(4):1019-23

1  Parasitology Section, Animal Health Department, Veterinary Faculty, University of Extremadura, Avenida de la Universidad s/n, 10071, Cáceres, Spain.

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.
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http://dx.doi.org/10.7589/2013-01-024DOI Listing
October 2013

Seroprevalence of Oestrus ovis (Diptera, Oestridae) infestation and associated risk factors in ovine livestock from southwestern Spain.

J Med Entomol 2005 May;42(3):327-31

Parasitology Section, Department of Medicine and Animal Health, Faculty of Veterinary Sciences, University of Extremadura, 10071 Cáceres, Spain.

This survey was carried out to determine the seroprevalence of nasal infestation by sheep bot fly, Oestrus ovis L., and to identify the risk factors associated with the disease in flocks in southwestern Spain. In total, 5,878 sera samples of adult sheep were collected at random in 551 farms from four provinces in the southwestern Spain: Badajoz, Cáceres, Córdoba, and Sevilla. Sera were tested by enzyme-linked immunosorbent assay for O. ovis antibodies, by using a crude L2 larval as antigen. The seropositive mean prevalence was 69.30%, and mean percentage of optical densities was 61.83%. There were significant differences between the provinces studied; C6rdoba and Sevilla were the provinces with more infested animals and higher seroprevalences. The correlation between seroprevalence and percentage of antibodies by farms was significant. There were only 18 farms free of seropositive animals, and 115 of the total 551 farms had all sampled animals seropositive, an indication of the high importance of this parasitosis in the investigated areas. Altitude, latitude, flock size, and ovine population density were the potential risk factors associated with the detection of O. ovis antibodies. Those animals breeding in regions located at low altitudes (<500 m), meridian latitudes (<39.5 degrees N), and on farms with medium-to-large flock size (>250 sheep) and high ovine population density (>100 sheep per km2) were more likely to be seropositive. These findings confirm that these studied factors should be considered as potential risk factors to the presence of O. ovis in ovines from southwestern Spain.
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http://dx.doi.org/10.1093/jmedent/42.3.327DOI Listing
May 2005
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