Publications by authors named "Rafael Calero-Bernal"

63 Publications

Diarrhoea-causing enteric protist species in intensively and extensively raised pigs (Sus scrofa domesticus) in Southern Spain. Part II: Association with Hepatitis E virus susceptibility.

Transbound Emerg Dis 2021 Nov 30. Epub 2021 Nov 30.

Parasitology Reference and Research Laboratory, National Centre for Microbiology, Ctra. Majadahonda-Pozuelo Km 2, Majadahonda, Madrid, 28220, Spain.

Enteropathogenic parasites can infect a wide range of mammals, including humans, supposing an important zoonotic risk. Hepatitis E virus (HEV) is an emerging foodborne pathogen of increasing public health relevance, affecting both human and animal populations. Because both microorganisms share faecal-oral transmission route they may constitute an excellent model to evaluate the interplay between them. Thus, we aim to evaluate the viral-parasite interactions at the enteric interface in swine. We included pigs of two different breeds farming in South Spain under different production systems. We compared the HEV prevalence by the presence of Giardia duodenalis, Cryptosporidium spp., Balantioides coli, Blastocystis sp., and Enterocytozoon bieneusi in faecal samples. The HEV prevalence was 13.1 (62 out 475, 95% CI: 10.2-16.4). Those pigs infected with Cryptosporidium spp. showed a higher prevalence of HEV (30.8% vs. 12%; p = 0.012). In the same way, animals bearing E. bieneusi seem to have a higher rate of HEV infection (24.2% vs. 12.2%; p = 0.06). According to their location in the gut, animals bearing intracellular enteroparasites showed a higher HEV prevalence than those uninfected (29.6% vs. 12.7%; p = 0.038), meanwhile those carrying extracellular enteroparasites had a lower likelihood to be infected by HEV than those uninfected (12.1% vs. 23.1%; p = 0.071). Those animals bearing both type of enteroparasites showed a similar prevalence of HEV infection than those exhibiting negative for both (20.8% vs. 26.1%; p = 0.763). Our study provides evidence that intracellular and extracellular enteroparasites modulate the susceptibility to HEV infection in pigs. Meanwhile, the presence of extracellular enteroparasites shows a protective effect on the risk of HEV acquisition in swine, whereas intracellular enteroparasites seems to have the opposite effect, favouring the HEV infection. This article is protected by copyright. All rights reserved.
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http://dx.doi.org/10.1111/tbed.14408DOI Listing
November 2021

Molecular Detection and Characterization of Intestinal and Blood Parasites in Wild Chimpanzees () in Senegal.

Animals (Basel) 2021 Nov 17;11(11). Epub 2021 Nov 17.

Parasitology Reference and Research Laboratory, Spanish National Centre for Microbiology, 28220 Majadahonda, Spain.

Wild chimpanzee populations in West Africa () have dramatically decreased as a direct consequence of anthropogenic activities and infectious diseases. Little information is currently available on the epidemiology, pathogenic significance, and zoonotic potential of protist species in wild chimpanzees. This study investigates the occurrence and genetic diversity of intestinal and blood protists as well as filariae in faecal samples ( = 234) from wild chimpanzees in the Dindefelo Community Nature Reserve, Senegal. PCR-based results revealed the presence of intestinal potential pathogens ( spp.: 11.5%; : 2.1%; : 0.9%), protist of uncertain pathogenicity ( sp.: 5.6%), and commensal species (: 18.4%; : 5.6%). , , and were undetected. Blood protists including (0.4%), (1.3%), and (9.8%) were also identified. Sanger sequencing analyses revealed host-adapted genetic variants within , but other parasitic pathogens (, , , ) have zoonotic potential, suggesting that cross-species transmission between wild chimpanzees and humans is possible in areas where both species overlap. Additionally, we explored potential interactions between intestinal/blood protist species and seasonality and climate variables. Chimpanzees seem to play a more complex role on the epidemiology of pathogenic and commensal protist and nematode species than initially anticipated.
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http://dx.doi.org/10.3390/ani11113291DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8614354PMC
November 2021

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

Diarrhoea-causing enteric protist species in intensively and extensively raised pigs (Sus scrofa domesticus) in Southern Spain. Part I: Prevalence and genetic diversity.

Transbound Emerg Dis 2021 Nov 9. Epub 2021 Nov 9.

Parasitology Reference and Research Laboratory, National Centre for Microbiology, Madrid, Spain.

Numerous protist species are shared between humans and pigs. Among those, Giardia duodenalis, Cryptosporidium spp. and Balantioides coli have a clear public and animal health significance. For others such as Enterocytozoon bieneusi and Blastocystis sp., their impact on animal health has not been fully established. Little information is currently available on the molecular diversity of these protists in swine populations. To fill this gap, we molecularly assessed G. duodenalis, Cryptosporidium spp., B. coli, Blastocystis sp. and E. bieneusi in faecal samples from Iberian and Large White pigs raised under different (intensive and/or extensive) management systems in southern Spain. A total of 151 extensively raised Iberian pigs, 140 intensively raised Iberian pigs, and 184 intensively raised Large White pigs were investigated. Blastocystis sp. was the agent most prevalently found (47.8%), followed by B. coli (45.5%), G. duodenalis (10.7%), E. bieneusi (6.9%), and Cryptosporidium spp. (5.5%). Blastocystis sp. was significantly less prevalent in intensively raised Iberian pigs (22.9%) than in their extensively raised counterparts (51.0%) or in intensively raised Large White pigs (64.1%). A significantly higher prevalence was found for G. duodenalis, Cryptosporidium spp., and E. bieneusi in Large White pigs than Iberian pigs. Balantioides coli was similarly distributed (40.0-51.1%) in all three investigated swine populations. Sequence analyses revealed the presence of G. duodenalis assemblage E, two Cryptosporidium species (Cryptosporidium scrofarum and Cryptosporidium suis), B. coli (genotypes A and B), Blastocystis sp. (ST1, ST3, and ST5), and E. bieneusi (EbpA, EbpC, EbpD, O, and a novel genotype named PigSpEb2). Novel genotype PigSpEb2 was found alone or in combination with EbpA. Data suggest a widespread exposure to protist enteroparasites in domestic pig populations irrespectively of breed and raising management system. Many of the species/genotypes identified have a zoonotic potential and might represent a public health concern.
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http://dx.doi.org/10.1111/tbed.14388DOI Listing
November 2021

First Description of Sarcoptic Mange in a Free-Ranging European Wildcat () from Spain.

Animals (Basel) 2021 Aug 25;11(9). Epub 2021 Aug 25.

Department of Animal Physiology, Faculty of Veterinary Medicine, Complutense University of Madrid, 28040 Madrid, Spain.

Sarcoptic mange caused by the mite is a worldwide-distributed skin infestation with a wide range of hosts, among them several species within the Felidae family. was diagnosed in a dead adult female European wildcat () from Spain, based on histological evaluation of skin biopsies and identification of the arthropod from skin scrapings and molecular methods. This is the first description of in a European wildcat. Due to its critical demography in the southernmost population of the Iberian Peninsula, the impacts of infectious diseases, including sarcoptic mange, as a new potential threat should be considered during disease surveillance programs of the species' populations.
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http://dx.doi.org/10.3390/ani11092494DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8466406PMC
August 2021

Molecular Detection and Characterization of sp. and in Cattle in Northern Spain.

Vet Sci 2021 Sep 11;8(9). Epub 2021 Sep 11.

Parasitology Reference and Research Laboratory, National Centre for Microbiology, Majadahonda, 28220 Madrid, Spain.

Some enteric parasites causing zoonotic diseases in livestock have been poorly studied or even neglected. This is the case in stramenopile sp. and the microsporidia in Spain. This transversal molecular epidemiological survey aims to estimate the prevalence and molecular diversity of sp. and in cattle faecal samples ( = 336) in the province of Álava, Northern Spain. Initial detection of and was carried out by polymerase chain reaction (PCR) and Sanger sequencing of the small subunit () rRNA gene and internal transcribed spacer (ITS) region, respectively. Intra-host subtype diversity was further investigated by next generation amplicon sequencing (NGS) of the rRNA gene in those samples that tested positive by conventional PCR. Amplicons compatible with sp. and were observed in 32.1% (108/336, 95% CI: 27.2-37.4%) and 0.6% (2/336, 95% CI: 0.0-1.4%) of the cattle faecal samples examined, respectively. Sanger sequencing produced ambiguous/unreadable sequence data for most of the isolates sequenced. NGS allowed the identification of 10 subtypes including ST1, ST3, ST5, ST10, ST14, ST21, ST23, ST24, ST25, and ST26. All -positive isolates involved mixed infections of 2-8 STs in a total of 31 different combinations. The two sequences were confirmed as potentially zoonotic genotype BEB4. Our data demonstrate that mixed subtype infections are extremely frequent in cattle in the study area. NGS was particularly suited to discern underrepresented subtypes or mixed subtype infections that were undetectable or unreadable by Sanger sequencing. The presence of zoonotic ST1, ST3, and ST5, and BEB4 suggest cross-species transmission and a potential risk of human infection/colonization.
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http://dx.doi.org/10.3390/vetsci8090191DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8473172PMC
September 2021

First description of Sarcocystis species infecting Barbary sheep (Ammotragus lervia).

Parasitol Res 2021 Aug 12;120(8):2881-2886. Epub 2021 Jul 12.

SALUVET Group, Animal Health Department, Complutense University of Madrid, Avda. Complutense s/n, 28040, Madrid, Spain.

Barbary sheep (Ammotragus lervia) is a North African native wild Caprinae, introduced in the 1970s in new territories such as Spain, the USA, and Mexico. Here, we describe Sarcocystis species in Barbary sheep. Sarcocysts were found in 19 out of 56 adult A. lervia in Southern Spain and characterized morphologically and molecularly. By light microscopy, sarcocysts had thin (< 1 μm) or thick (> 2 μm) walls. By transmission electron microscopy, sarcocysts with thick walls had Type 14 villar protrusions corresponding to S. tenella/S. capracanis of domestic sheep (Ovis aries) or goats (Capra hircus). Sarcocysts with thin walls had Type 7b villar protrusions that corresponded to S. arieticanis/S. hircicanis of domestic sheep or goats. Molecular analyses allowed the identification of only thick-walled Sarcocystis species. Six sarcocysts were assigned to S. tenella (99.2-100% and 95.6-100% sequence similarity within 18S rRNA and COI, respectively) and 19 sarcocysts were assigned to S. capracanis (98.5-99.8% and 97.9-99.0% sequence similarity within 18S rRNA and COI, respectively). Further studies are needed for taxonomic identification of sarcocysts in Barbary sheep because Sarcocystis species in sheep and goats are not cross transmissible despite morphological similarities.
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http://dx.doi.org/10.1007/s00436-021-07239-zDOI Listing
August 2021

Molecular survey of Besnoitia spp. (Apicomplexa) in faeces from European wild mesocarnivores in Spain.

Transbound Emerg Dis 2021 Nov 7;68(6):3156-3166. Epub 2021 Jul 7.

SALUVET, Department of Animal Health, Faculty of Veterinary, Complutense University of Madrid, Madrid, Spain.

Numerous studies have unsuccessfully tried to unravel the definitive host of the coccidian parasite Besnoitia besnoiti. Cattle infections by B. besnoiti cause a chronic and debilitating condition called bovine besnoitiosis that has emerged in Europe during the last two decades, mainly due to limitations in its control associated with the absence of vaccines and therapeutical tools. Although the exact transmission pathways of B. besnoiti is currently unknown, it is assumed that the parasite might have an indirect life cycle with a carnivore as definitive host. Current lack of studies in wildlife might underestimate the importance of free-living species in the epidemiology of B. besnoiti. Thus, the aim of the present study is to assess the presence of Besnoitia spp. in free-ranging mesocarnivores in Spain. DNA was searched by PCR on faeces collected from wild carnivores as a first approach to determine which species could be considered as potential definitive host candidates in further research. For this purpose, a total of 352 faecal samples from 12 free-living wild carnivore species belonging to the Canidae, Felidae, Herpestidae, Mustelidae, Procyonidae and Viverridae families were collected in seven Spanish regions. PCR testing showed that Besnoitia spp. DNA was present in four faecal samples from red foxes collected in western Spain, an area with the greatest density of extensively reared cattle and associated with high incidence of bovine besnoitiosis in the country. To date, this is the first report of a B. besnoiti-like sequence (99.57% homology) from carnivore faeces in a worldwide context. Red foxes might contribute to the epidemiology of B. besnoiti, although further studies, mostly based on bioassay, would be needed to elucidate the accuracy and extent of these interesting findings.
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http://dx.doi.org/10.1111/tbed.14206DOI Listing
November 2021

In vivo and in vitro models show unexpected degrees of virulence among Toxoplasma gondii type II and III isolates from sheep.

Vet Res 2021 Jun 10;52(1):82. Epub 2021 Jun 10.

SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Madrid, Spain.

Toxoplasma gondii is an important zoonotic agent with high genetic diversity, complex epidemiology, and variable clinical outcomes in animals and humans. In veterinary medicine, this apicomplexan parasite is considered one of the main infectious agents responsible for reproductive failure in small ruminants worldwide. The aim of this study was to phenotypically characterize 10 Spanish T. gondii isolates recently obtained from sheep in a normalized mouse model and in an ovine trophoblast cell line (AH-1) as infection target cells. The panel of isolates met selection criteria regarding such parameters as genetic diversity [types II (ToxoDB #1 and #3) and III (#2)], geographical location, and sample of origin (aborted foetal brain tissues or adult sheep myocardium). Evaluations of in vivo mortality, morbidity, parasite burden and histopathology were performed. Important variations between isolates were observed, although all isolates were classified as "nonvirulent" (< 30% cumulative mortality). The isolates TgShSp16 (#3) and TgShSp24 (#2) presented higher degrees of virulence. Significant differences were found in terms of in vitro invasion rates and tachyzoite yield at 72 h post-inoculation (hpi) between TgShSp1 and TgShSp24 isolates, which exhibited the lowest and highest rates, respectively. The study of the CS3, ROP18 and ROP5 loci allelic profiles revealed only type III alleles in ToxoDB #2 isolates and type II alleles in the #1 and #3 isolates included. We concluded that there are relevant intra- and inter-genotype virulence differences in Spanish T. gondii isolates, which could not be inferred by genetic characterization using currently described molecular markers.
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http://dx.doi.org/10.1186/s13567-021-00953-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8194156PMC
June 2021

Long-Term Preservation and Storage of Faecal Samples in Whatman Cards for PCR Detection and Genotyping of and .

Animals (Basel) 2021 May 12;11(5). Epub 2021 May 12.

Parasitology Reference and Research Laboratory, Spanish National Centre for Microbiology, Majadahonda, 28220 Madrid, Spain.

Preservation and conservation of biological specimens, including faecal samples, is a challenge in remote areas or poor-resource settings where the cold chain cannot be maintained. This study aims at evaluating the suitability of filter cards for long-term storage of faecal samples of animal and human origin positive to the diarrhoea-causing protozoan parasites, and . Three commercially available Whatman Filter Cards were comparatively assessed: the FTA Classic Card, the FTA Elute Micro Card, and the 903 Protein Saver Card. Human faecal samples positive to ( = 5) and ( = 5) were used to impregnate the selected cards at given storage (1 month, 3 months, and 6 months) periods and temperature (-20 °C, 4 °C, and room temperature) conditions. Parasite DNA was detected by PCR-based methods. Sensitivity assays and quality control procedures to assess suitability for genotyping purposes were conducted. Overall, all three Whatman cards were proven useful for the detection and molecular characterisation of and under the evaluated conditions. Whatman cards represent a simple, safe, and cost-effective option for the transportation, preservation, and storage of faecal samples without the need of the cold chain.
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http://dx.doi.org/10.3390/ani11051369DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8151430PMC
May 2021

Toxoplasma gondii infection in domestic and wild felids as public health concerns: a systematic review and meta-analysis.

Sci Rep 2021 05 4;11(1):9509. Epub 2021 May 4.

Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Felidae as definitive hosts for Toxoplasma gondii play a major role in transmission to all warm-blooded animals trough oocysts dissemination. Therefore the current comprehensive study was performed to determine the global status of T. gondii infection in domestic and wild felids aiming to provide comprehensive data of interest for further intervention approaching the One Health perspective. Different databases were searched by utilizing particular key words for publications related to T. gondii infecting domestic and wild feline host species, worldwide, from 1970 to 2020. The review of 337 reports showed that the seroprevalence of T. gondii in domestic cats and wild felids was estimated in 37.5% (95% CI 34.7-40.3) (I = 98.3%, P < 0.001) and 64% (95% CI 60-67.9) (I = 88%, P < 0.0001), respectively. The global pooled prevalence of oocysts in the fecal examined specimens from domestic cats was estimated in 2.6% (95% CI 1.9-3.3) (I = 96.1%, P < 0.0001), and that in fecal samples from wild felids was estimated in 2.4% (95% CI 1.1-4.2) (I = 86.4%, P < 0.0001). In addition, from 13,252 examined soil samples in 14 reviewed studies, the pooled occurrence of T. gondii oocysts was determined in 16.2% (95% CI 7.66-27.03%). The observed high rates of anti-T. gondii antibodies seroprevalence levels and oocyst excretion frequency in the felids, along with soil (environmental) contamination with oocysts may constitute a potential threat to animal and public health, and data will result of interest in further prophylaxis programs.
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http://dx.doi.org/10.1038/s41598-021-89031-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8097069PMC
May 2021

Evidence for Unknown -Like Infection in Stranded Striped Dolphins () from the Ligurian Sea, Italy.

Animals (Basel) 2021 Apr 22;11(5). Epub 2021 Apr 22.

Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, 10154 Torino, Italy.

Two striped dolphins (SD1, SD2), stranded along the Ligurian coast of Italy, were diagnosed with a nonsuppurative meningoencephalitis associated with previously undescribed protozoan tissue cysts. As tissue cysts were morphologically different from those of , additional histopathological, immunohistochemical, ultrastructural, and biomolecular investigations were performed, aiming to fully characterize the organism. Histopathology revealed the presence of large -like tissue cysts, associated with limited inflammatory lesions in all CNS areas studied. IHC was inconclusive, as positive staining with polyclonal antisera did not preclude cross-reaction with other Sarcocystidae coccidia. Applied to each animal, 11 different PCR protocols precluded a neural infection by , and . coinfection was confirmed only in dolphin SD2. sp. sequences, showing the highest homology to species infecting the Bovidae family, were amplified from SD1 myocardium and SD2 skeletal muscle. The present study represents the first report of -like tissue cysts in the brain of stranded cetaceans along with the first description of sp. infection in muscle tissue of dolphins from the Mediterranean basin.
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http://dx.doi.org/10.3390/ani11051201DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8143450PMC
April 2021

Occurrence and Genetic Diversity of Protist Parasites in Captive Non-Human Primates, Zookeepers, and Free-Living Sympatric Rats in the Córdoba Zoo Conservation Centre, Southern Spain.

Animals (Basel) 2021 Mar 5;11(3). Epub 2021 Mar 5.

Parasitology Reference and Research Laboratory, Spanish National Centre for Microbiology, 28220 Madrid, Spain.

Little information is currently available on the epidemiology of parasitic and commensal protist species in captive non-human primates (NHP) and their zoonotic potential. This study investigates the occurrence, molecular diversity, and potential transmission dynamics of parasitic and commensal protist species in a zoological garden in southern Spain. The prevalence and genotypes of the main enteric protist species were investigated in faecal samples from NHP ( = 51), zookeepers ( = 19) and free-living rats ( = 64) by molecular (PCR and sequencing) methods between 2018 and 2019. The presence of spp. was also investigated in tissues from sympatric rats using PCR. sp. (45.1%), (27.5%), (21.6%), (3.9%), and (2.0%) (but not spp.) were detected in NHP. (10.5%) and sp. (10.5%) were identified in zookeepers, while spp. (45.3%), (14.1%), and sp. (6.25%) (but not spp.) were detected in rats. ST1, ST3, and ST8 and sub-assemblage AII were identified in NHP, and ST1 in zookeepers. isolates failed to be genotyped in human samples. In rats, four (, , and rat genotypes IV and V), one (assemblage G), and three (ST4) genetic variants were detected. Our results indicate high exposure of NHP to zoonotic protist species. Zoonotic transmission of ST1 was highly suspected between captive NHP and zookeepers.
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http://dx.doi.org/10.3390/ani11030700DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8035673PMC
March 2021

Parasites of the Reintroduced Iberian Lynx () and Sympatric Mesocarnivores in Extremadura, Spain.

Pathogens 2021 Mar 1;10(3). Epub 2021 Mar 1.

Asistencia Técnica de la Dirección General del Medio Natural y Desarrollo Sostenible de la Junta de Comunidades de Castilla-La Mancha, Plaza del Cardenal Siliceo s/n, 45071 Toledo, Spain.

The Iberian lynx () is one of the most endangered felid species in the world. Conservation efforts have increased its population size and distribution and reinforced their genetic diversity through captive breeding and reintroduction programmes. Among several threats that the Iberian lynx faces, infectious and parasitic diseases have underlined effects on the health of their newly reintroduced populations, being essential to identify the primary sources of these agents and assess populations health status. To achieve this, 79 fresh faecal samples from Iberian lynx and sympatric mesocarnivores were collected in the reintroduction area of Extremadura, Spain. Samples were submitted to copromicroscopic analyses to assess parasite diversity, prevalence, and mean intensity of parasite burden. Overall, 19 (24.1%, ±15.1-35.0) samples were positive for at least one enteric parasite species. Parasite diversity and prevalence were higher in the Iberian lynx (43.8%) compared with the others mesocarnivores under study (e.g., the red fox and the Egyptian mongoose ). Ancylostomatidae and were the most prevalent (15.6%) parasites. Obtained results revealed that Iberian lynx role as predator control might have reduced parasite cross-transmission between this felid and mesocarnivores due to their decreasing abundances. Surveillance programs must include regular monitoring of this endangered felid, comprising mesocarnivores, but also domestic/feral and wild cat communities.
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http://dx.doi.org/10.3390/pathogens10030274DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8000845PMC
March 2021

Multilocus Genotyping of in Mostly Asymptomatic Indigenous People from the Tapirapé Tribe, Brazilian Amazon.

Pathogens 2021 Feb 14;10(2). Epub 2021 Feb 14.

Parasitology Reference and Research Laboratory, Spanish National Centre for Microbiology, Majadahonda, 28220 Madrid, Spain.

Little information is available on the occurrence and genetic variability of the diarrhoea-causing enteric protozoan parasite in indigenous communities in Brazil. This cross-sectional epidemiological survey describes the frequency, genotypes, and risk associations for this pathogen in Tapirapé people (Brazilian Amazon) at four sampling campaigns during 2008-2009. Microscopy was used as a screening test, and molecular (PCR and Sanger sequencing) assays targeting the small subunit ribosomal RNA, the glutamate dehydrogenase, the beta-giardin, and the triosephosphate isomerase genes as confirmatory/genotyping methods. Associations between and sociodemographic and clinical variables were investigated using Chi-squared test and univariable/multivariable logistic regression models. Overall, 574 individuals belonging to six tribes participated in the study, with prevalence rates varying from 13.5-21.7%. The infection was positively linked to younger age and tribe. Infected children <15 years old reported more frequent gastrointestinal symptoms compared to adults. Assemblage B accounted for three out of four infections and showed a high genetic diversity. No association between assemblage and age or occurrence of diarrhoea was demonstrated. These data indicate that the most likely source of infection was anthropic and that different pathways (e.g., drinking water) may be involved in the transmission of the parasite.
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http://dx.doi.org/10.3390/pathogens10020206DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7917967PMC
February 2021

Detection of Myxoma Virus DNA in Ticks from Lagomorph Species in Spain Suggests Their Possible Role as Competent Vector in Viral Transmission.

J Wildl Dis 2021 04;57(2):423-428

SALUVET Group, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Avda. Puerta de Hierro s/n, 28040, Madrid, Spain.

Myxoma virus (MYXV) causes morbidity and mortality in European wild rabbits (Oryctolagus cuniculus) worldwide, and recently in Iberian hares (Lepus granatensis) in Spain. We aimed to assess the presence of MYXV-specific DNA in ixodid ticks collected from both hosts. A total of 417 ticks harvested from 30 wild lagomorphs, including wild rabbits and Iberian hares were collected from southern Spain. Enzyme-linked immunosorbent assay and PCR-sequencing were used to detect virus exposure and presence, respectively. Antibodies to MYXV were detected in 68% (17/25) of wild rabbits and in 67% (2/3) of Iberian hares. We detected MYXV DNA in 50.7% of pools of two different tick species (nymphs and adults of Rhipicephalus pusillus, and nymphs of Hyalomma lusitanicum) parasitizing rabbits and hares. The obtained partial sequence of the viral major envelope protein gene showed a mutation (G383A) within the MYXV_gp026 locus between the rabbit strain and Iberian hare strain (recently isolated in tissues of infected hares from Spain). However, in our study, the viral DNA presence was detected for the first time using tick DNA as the PCR-template, but the possible role of ticks as vectors of MYXV still needs to be elucidated.
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http://dx.doi.org/10.7589/JWD-D-20-00116DOI Listing
April 2021

Microbial community of Hyalomma lusitanicum is dominated by Francisella-like endosymbiont.

Ticks Tick Borne Dis 2021 03 14;12(2):101624. Epub 2020 Dec 14.

SaBio. Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005, Ciudad Real, Spain; Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078, USA.

Exploring tick associations with complex microbial communities and single-microbial partners, especially intracellular symbionts, has become crucial to understand tick biology. Of particular interest are the underlying interactions with biological consequences i.e. tick fitness, vector competence. In this study, we first sequenced the 16S rRNA bacterial phylogenetic marker in adult male ticks of Hyalomma lusitanicum collected from 5 locations in the province of Cáceres to explore the composition of its microbial community. Overall, 16S rRNA sequencing results demonstrated that the microbial community of H. lusitanicum is mostly dominated by Francisella-like endosymbionts (FLEs) (ranging from 52% to 99% of relative abundance) suggesting it is a key taxon within the microbial community and likely a primary endosymbiont. However, further research is required to explore the mechanisms underlying the interaction between FLEs and H. lusitanicum.
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http://dx.doi.org/10.1016/j.ttbdis.2020.101624DOI Listing
March 2021

Isolation, Genotyping, and Mouse Virulence Characterization of From Free Ranging Iberian Pigs.

Front Vet Sci 2020 20;7:604782. Epub 2020 Nov 20.

Salud Veterinaria y Zoonosis (SALUVET) Group, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Madrid, Spain.

The present study aimed to isolate and perform molecular and phenotypic characterization of strains infecting Iberian pigs bred under semi-free conditions and destined for human consumption. Blood and heart tissue samples from 361 fattening pigs from 10 various herds selected in the main areas of Iberian pig production were collected at a slaughterhouse; the sera were tested for anti- antibodies using a commercial indirect ELISA kit, and a mouse bioassay was carried out using heart muscle of seropositive individual representatives from each geographical location. Seventy-nine (21.9%) of the 361 animals tested positive for anti- antibodies according to the serology test. Fifteen samples of myocardial tissue were subjected to bioassay and 5 isolates (TgPigSp1 to TgPigSp5) were obtained. The isolates were characterized by using 11 PCR-RFLP genetic markers; three isolates had a ToxoDB #3 genotype (3/5) and two isolates had a ToxoDB #2 genotype (2/5). The TgPigSp1 and TgPigSp4 isolates were selected for virulence in mice characterization as instances of each different RFLP-genotype found. The TgPigSp1 isolate (#2 genotype) was virulent in mice with notable cumulative mortality (87.5%) and morbidity rates (100%); the TgPigSp4 (#3) was nonvirulent and triggered mild clinical signs in 42.1% of seropositive mice. Infection dynamics and organ distribution of both isolates were analyzed; the data revealed significant differences, including substantially higher parasite load in the lung during the acute phase of infection, in mice infected with TgPigSp1 than in the case of TgPigSp4 (median parasite load 7.6 vs. 0 zoites/mg, respectively; < 0.05). Furthermore, degrees of severity of detected histopathological lesions appeared to be related to higher parasite burdens. Taking into account the unexpectedly high mortality rate and parasite load associated with the clonal genotype III, which is traditionally considered nonvirulent in mice, the need for further investigation and characterization of the strains circulating in any host in Europe is emphasized.
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http://dx.doi.org/10.3389/fvets.2020.604782DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7714755PMC
November 2020

Correction to: Isolation and genetic characterization of Toxoplasma gondii in Spanish sheep flocks.

Parasit Vectors 2020 Nov 3;13(1):545. Epub 2020 Nov 3.

SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040, Madrid, Spain.

An amendment to this paper has been published and can be accessed via the original article.
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http://dx.doi.org/10.1186/s13071-020-04423-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7640693PMC
November 2020

Isolation and genetic characterization of Toxoplasma gondii in Spanish sheep flocks.

Parasit Vectors 2020 Aug 5;13(1):396. Epub 2020 Aug 5.

SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040, Madrid, Spain.

Background: Toxoplasma gondii is a major cause of abortion in small ruminants and presents a zoonotic risk when undercooked meat containing cysts is consumed. The aim of the present study was to investigate the genetic diversity among the T. gondii strains circulating in ovine livestock in Spain.

Methods: Selected samples collected from abortion outbreaks due to toxoplasmosis (n = 31) and from chronically infected adult sheep at slaughterhouses (n = 50) in different Spanish regions were bioassayed in mice, aiming at parasite isolation. In addition, all original clinical samples and the resulting isolates were genotyped by multi-nested PCR-RFLP analysis of 11 molecular markers and by PCR-DNA sequencing of portions of the SAG3, GRA6 and GRA7 genes.

Results: As a result, 30 isolates were obtained from 9 Spanish regions: 10 isolates from abortion-derived samples and 20 isolates from adult myocardial tissues. Overall, 3 genotypes were found: ToxoDB#3 (type II PRU variant) in 90% (27/30) of isolates, ToxoDB#2 (clonal type III) in 6.7% (2/30), and ToxoDB#1 (clonal type II) in 3.3% (1/30). When T. gondii-positive tissue samples (n = 151) were directly subjected to RFLP genotyping, complete restriction profiles were obtained for 33% of samples, and up to 98% of the specimens belonged to the type II PRU variant. A foetal brain showed a clonal type II pattern, and four specimens showed unexpected type I alleles at the SAG3 marker, including two foetal brains that showed I + II alleles as co-infection events. Amplicons of SAG3, GRA6 and GRA7 obtained from isolates and clinical samples were subjected to sequencing, allowing us to confirm RFLP results and to detect different single-nucleotide polymorphisms.

Conclusions: The present study informed the existence of a predominant type II PRU variant genotype (ToxoDB#3) infecting domestic sheep in Spain, in both abortion cases and chronic infections in adults, coexisting with other clonal (ToxoDB#1 and ToxoDB#2), much less frequent genotypes, as well as polymorphic strains as revealed by clinical sample genotyping. The use of multilocus sequence typing aided in accurately estimating T. gondii intragenotype diversity.
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http://dx.doi.org/10.1186/s13071-020-04275-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7404076PMC
August 2020

Detection of new Crimean-Congo haemorrhagic fever virus genotypes in ticks feeding on deer and wild boar, Spain.

Transbound Emerg Dis 2021 May 11;68(3):993-1000. Epub 2020 Aug 11.

SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, Ciudad Real, Spain.

Crimean-Congo haemorrhagic fever virus (CCHFV) is the causative agent of the severe tick-borne, often fatal, zoonotic Crimean-Congo haemorrhagic fever (CCHF), which is widely distributed worldwide. The CCHFV transmission to humans occurs through tick bite, crushing of engorged ticks or contact with infected host blood. Previously, CCHFV genotype Africa III was reported in Spain. Given the emergence of CCHF and the role of ticks in pathogen maintenance and transmission, we investigated the presence and genotype identity of the virus in tick species parasitizing abundant wild host species in south-western Spain. A total of 613 adult ticks were collected from hunter-harvested wild ungulates in twenty locations throughout south-western Spain. Ticks were identified, nucleic acids were extracted, RNA was analysed by a nested RT-PCR targeting CCHFV S segment, and the amplicons were sequenced. According to the 212-bp sequence amplified, the presence of CCHFV human genotype Europe V was detected in Hyalomma lusitanicum and Dermacentor marginatus ticks collected from red deer, fallow deer and Eurasian wild boar in different locations from south-western Spain. Genotype Africa IV was also detected in a H. lusitanicum tick collected from a red deer. The detection of CCHFV in different tick species collected from various wild hosts and localities provided strong evidence of widespread CCHFV presence in the region, suggesting that the circulation of the virus in Spain requires more attention. Additionally, the identification of the CCHFV genotype Europe V in ticks suggested that its introduction in Spain was probably from Eastern Europe.
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http://dx.doi.org/10.1111/tbed.13756DOI Listing
May 2021

First identification of genotypes of Enterocytozoon bieneusi (Microsporidia) among symptomatic and asymptomatic children in Mozambique.

PLoS Negl Trop Dis 2020 06 30;14(6):e0008419. Epub 2020 Jun 30.

Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, Majadahonda, Madrid, Spain.

Enterocytozoon bieneusi is a human pathogen with a broad range of animal hosts. Initially, E. bieneusi was considered an emerging opportunistic pathogen in immunocompromised, mainly HIV-infected patients, but it has been increasingly reported in apparently healthy individuals globally. As in other African countries, the molecular epidemiology of E. bieneusi in Mozambique remains completely unknown. Therefore, we undertook a study to investigate the occurrence and genetic diversity of E. bieneusi infections in children with gastrointestinal symptoms as well as in asymptomatic children in Mozambique. Individual stool specimens were collected from 1,247 children aged between 0 and 14 years-old living in urban and rural settings in Zambézia (n = 1,097) and Maputo (n = 150) provinces between 2016 and 2019. Samples were analysed for E. bieneusi by nested-PCR targeting the internal transcribed spacer (ITS) region of the rRNA gene. All positive amplicons were confirmed and genotyped. Penalised logistic regression (Firth) was used to evaluate risk associations. The overall prevalence of E. bieneusi in this children population was 0.7% (9/1,247). A 10-fold higher prevalence was found in Maputo (4.0%; 6/150) than in Zambézia (0.3%; 3/1,097). All E. bieneusi-positive samples were from children older than 1-year of age, and most (8/9) from asymptomatic children. Nucleotide sequence analysis of the ITS region revealed the presence of four genotypes, three previously reported (Peru11, n = 1; Type IV, n = 2, and S2, n = 2) and a novel genotype (named HhMzEb1, n = 4). Novel genotype HhMzEb1 was identified in both asymptomatic (75%, 3/4) and symptomatic (25%, 1/4) children from a rural area in Maputo province in southern Mozambique. Genotypes HhMzEb1, Peru11, S2, and Type IV belonged to the Group 1 that includes genotypes with low host specificity and the potential for zoonotic and cross-species transmission. Being infected by enteric protozoan parasites and no handwashing were identified as risk associations for E. bieneusi infection. This study reports the first investigation of E. bieneusi genotypes in Mozambique with the identification of three previously reported genotypes in humans as well as a novel genotype (HhMzEb1). Findings highlight the need to conduct additional research to elucidate the epidemiology of E. bieneusi in the country, especially in rural areas where poor hygiene conditions still prevail. Special attention should be paid to the identification of suitable animal and environmental reservoirs of this parasite and to the characterization of transmission pathways.
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http://dx.doi.org/10.1371/journal.pntd.0008419DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7357779PMC
June 2020

Molecular survey for cyst-forming coccidia (Toxoplasma gondii, Neospora caninum, Sarcocystis spp.) in Mediterranean periurban micromammals.

Parasitol Res 2020 Aug 25;119(8):2679-2686. Epub 2020 Jun 25.

SALUVET Group, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040, Madrid, Spain.

Rodents and other micromammals constitute important reservoirs of infectious diseases; their role in the life cycle of apicomplexan parasites such as Toxoplasma gondii, Neospora caninum, and Sarcocystis spp. still needs clarification. In the present study, we analyzed by PCR and Sanger sequencing methods the presence of specific parasite DNA within brain and heart tissues of 313 individuals of five synanthropic small mammal species (Apodemus sylvaticus, Mus spretus, M. musculus, Rattus rattus, and Crocidura russula) collected in Barcelona metropolitan area (NE Spain). In addition, PCR-RFLP and microsatellites were also used as tools for genotypic characterization of T. gondii and N. caninum, respectively. Specific DNA of T. gondii, N. caninum, and Sarcocystis spp. was detected in 0.3% (n = 1), 1.3% (n = 4), and 3.8% (n = 12) of the animals, respectively. No mixed infections were observed. Crocidura russula stood out as the main host for Sarcocystis spp. Toxoplasma gondii-specific DNA detected in a house rat was genetically characterized by PCR-RFLP, presenting type II and III alleles (SAG1 [II], SAG3 [II], GRA6 [II], c22-8 [III], Apico [III]). Also, unsuccessful DNA sequencing and microsatellite typing were attempted in N. caninum-positive samples, which suggested a lack of PCR specificity and open avenues to speculate the host competence of rodents for N. caninum. Likewise, Sarcocystis spp. identity was studied by alignment and phylogenetic analyses of cox1 and 28S rRNA sequences from the 14 positive samples. It resulted in at least three unknown organisms closely similar (95.7-100% cox1-sequence homology) to Sarcocystis pantherophisi from the Eastern rat snake (Pantherophis alleghaniensis) (KU891603), suggesting together with 28S rRNA sequences analyses, three Sarcocystis sp. with a life cycle conformed by rodents as intermediate host (IH) and snakes as definitive hosts (DH) infecting the periurban micromammals surveyed. Prevalence figures found in this first survey carried out in Spain agree with other international studies focused on periurban areas. Further surveys should be conducted in farms and their surroundings in order to unravel the role of wild micromammals in the epidemiology of such protozoan parasites affecting our livestock, and therefore human population.
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http://dx.doi.org/10.1007/s00436-020-06777-2DOI Listing
August 2020

Enterocytozoon bieneusi (Microsporidia): Identification of novel genotypes and evidence of transmission between sympatric wild boars (Sus scrofa ferus) and Iberian pigs (Sus scrofa domesticus) in Southern Spain.

Transbound Emerg Dis 2020 Nov 15;67(6):2869-2880. Epub 2020 Jun 15.

Parasitology Reference and Research Laboratory, National Centre for Microbiology, Madrid, Spain.

Microsporidia is a phylum of obligate emergent intracellular protist-like fungi pathogens that infect a broad range of hosts including vertebrates and invertebrates. Enterocytozoon bieneusi is the most common cause of microsporidiosis in humans, affecting primarily immunosuppressed patients but also reported in immunocompetent individuals. Epidemiological information on the presence and molecular diversity of E. bieneusi in livestock and wildlife in Spain is limited. Therefore, the occurrence of this microsporidia was investigated in sympatric extensively reared Iberian pigs (n = 186) and free ranging wild boars (n = 142) in the province of Córdoba, Southern Spain. Forty-two Iberian pigs (22.6%) and three wild boars (2.1%) were found E. bieneusi positive by PCR. In Iberian pigs, occurrence of E. bieneusi was significantly higher in sows than in fattening pigs (31.6% vs. 11.4%; p = .001). Five genotypes were identified in Iberian pigs, four previously reported (EbpA, PigEb4, O, Pig HN-II) and a novel genotype (named PigSpEb1), while only two genotypes were identified in wild boars, EbpA and novel genotype PigSpEb1. All five genotypes identified belong to Group 1 suggesting zoonotic potential. This study constitutes the first report on the occurrence and molecular characterization of E. bieneusi in Iberian pigs and wild boars. The identification of two genotypes with zoonotic potential in sympatric Iberian pigs and wild boars suggests that E. bieneusi can be potentially transmitted between those two hosts, but also implies that they may act as natural sources of microsporidia infection to other hosts including humans.
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http://dx.doi.org/10.1111/tbed.13658DOI Listing
November 2020

Protist enteroparasites in wild boar (Sus scrofa ferus) and black Iberian pig (Sus scrofa domesticus) in southern Spain: a protective effect on hepatitis E acquisition?

Parasit Vectors 2020 Jun 3;13(1):281. Epub 2020 Jun 3.

Parasitology Reference and Research Laboratory, Spanish National Centre for Microbiology, Majadahonda, Madrid, Spain.

Background: Several studies have independently evaluated the occurrence of hepatitis E virus (HEV) and enteroparasites in swine, but no surveys have been conducted to jointly assess the prevalence and genetic diversity of enteroparasites in pigs and wild boars, their sympatric transmission between hosts, and their potential interaction with HEV.

Methods: We prospectively collected serum and faecal samples from black Iberian domestic pigs and wild boars from southern Spain between 2015‒2016. We evaluated for HEV in serum and faeces, and for the presence of enteroparasites (Giardia duodenalis, Cryptosporidium spp., Blastocystis sp., Neobalantidium coli and Strongyloides spp.) in the same faecal samples. The prevalence of each intestinal parasite species was calculated.

Results: A total of 328 animals (56.7% black Iberian pigs and 43.3% wild boars) were included in the study. The overall global prevalence of HEV in serum was 16.8%. The overall global prevalence of each enteroparasite species was 19.5% for G. duodenalis, 8.2% for Cryptosporidium spp., 41.8% for Blastocystis sp., 31.4% for N. coli, and 8.8% for Strongyloides spp. HEV-infected animals showed a significantly lower prevalence of G. duodenalis (3.2 vs 20%; P = 0.002) and Blastocystis sp. (38.7 vs 80%; P < 0.001) than those uninfected by HEV. Animals carrying G. duodenalis and Blastocystis sp. infections showed a significantly lower rate of HEV infection than those not harbouring these enteroparasites (P < 0.001).

Conclusions: Our study found a high prevalence of enteroparasites in black Iberian pigs and wild boars in southern Spain, suggesting a sympatric co-transmission of some of the species investigated. It is suggested that extracellular G. duodenalis and Blastocystis sp. might have a protective effect on HEV acquisition in swine.
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http://dx.doi.org/10.1186/s13071-020-04152-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7271453PMC
June 2020

First report of Thelazia callipaeda in a free-ranging Iberian wolf (Canis lupus signatus) from Spain.

Parasitol Res 2020 Jul 3;119(7):2347-2350. Epub 2020 Jun 3.

SALUVET, Department of Animal Health, Faculty of Veterinary, Complutense University of Madrid, Av. Puerta de Hierro s/n, 28040, Madrid, Spain.

Thelazia callipaeda (Spirurida, Thelaziidae) is a vector-borne zoonotic eyeworm able to infect a broad spectrum of carnivores. Here, we describe the first case of bilateral infection by T. callipaeda in the eyes of an adult female Iberian wolf (Canis lupus signatus) in central Spain. Nematodes collected were morphologically identified (n = 42), and two specimens were molecularly characterized. At the sequence analysis of the partial mitochondrial cytochrome c oxidase subunit 1 gene, T. callipaeda haplotype 1 (the only haplotype circulating in Europe) was detected. The role of the Iberian wolf as a natural reservoir for T. callipaeda in the life cycle of this emerging zoonosis and the implications in conservation are discussed.
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http://dx.doi.org/10.1007/s00436-020-06735-yDOI Listing
July 2020

Blastocystis sp. Subtype Diversity in Wild Carnivore Species from Spain.

J Eukaryot Microbiol 2020 03 26;67(2):273-278. Epub 2019 Nov 26.

Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, Ctra. Majadahonda-Pozuelo Km 2, 28220, Majadahonda, Madrid, Spain.

The occurrence and molecular diversity of the stramenopile eukaryote Blastocystis sp. was investigated by PCR and sequencing (Sanger and NGS) methods in 380 faecal specimens of free-living carnivores in Spain. Blastocystis sp. was confirmed in 1.6% (6/380) of the specimens analysed. Two samples from a common genet and a fox were successfully subtyped as ST7 by Sanger. Using NGS, ST14 was found in a fox and a European polecat, ST7 in a fox, and two additional foxes presented mixed infections of ST1/ST2/ST4 and ST1/ST2/ST7, respectively. Wild carnivore species could act as carriers of zoonotic Blastocystis subtypes.
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http://dx.doi.org/10.1111/jeu.12772DOI Listing
March 2020

(Apicomplexa) in Fallow Deer () from Spain: Ultrastructure and New Host Record.

J Parasitol 2019 10;105(5):813-815

SALUVET, Animal Health Department, Complutense University of Madrid, Avenida Complutense s/n, 28040, Madrid, Spain.

Members of the genus are frequently found infecting members of the family Cervidae. Although species are generally host specific for their intermediate hosts, species in cervids appear to be less host specific. Here, we report fallow deer () as a new host for , originally described from the red deer (). Tongues of 69 legally hunted animals in Spain were tested for sarcocysts, and the species were characterized by light microscopy, ultrastructurally and molecularly. Sarcocysts were identified in 66.7% of . Sarcocysts had thin (<2 μm thick) cyst wall with hair-like villar protrusions bifurcated at their tips resembling type 8a. Genetic sequences obtained for and reached 99.6-100% and 97.9-98.7% similarity, respectively, to those of from the red deer. The present study provides new data concerning lower level of host specificity within genus for cervids.
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October 2019

Occurrence and genetic diversity of Enterocytozoon bieneusi (Microsporidia) in owned and sheltered dogs and cats in Northern Spain.

Parasitol Res 2019 Oct 21;118(10):2979-2987. Epub 2019 Aug 21.

Parasitology Reference and Research Laboratory, National Centre for Microbiology, Majadahonda, Madrid, Spain.

Enterocytozoon bieneusi is an obligate intracellular protist-like fungi parasite that infects numerous mammal hosts including humans, raising concerns of zoonotic transmission. There is little information available on the presence and diversity of E. bieneusi genotypes in companion animals. Here, we determined the occurrence and genetic diversity of E. bieneusi in domestic dogs and cats from Northern Spain. A total of 336 genomic DNA samples extracted from canine (n = 237) and feline (n = 99) faecal specimens were retrospectively investigated. The presence of E. bieneusi was assessed by PCR of the rRNA internal transcribed spacer (ITS) gene. The parasite was detected in 3.0% (3/99) and 0.8% (2/237) of the cats and dogs examined, respectively. All three feline positive samples were from stray cats living in an urban setting, whereas the two canine samples were from owned dogs living in rural areas. Sequence analysis revealed the presence of two genotypes in dogs, BEB6 and PtEb IX, and two genotypes in cats, D and Peru11. The identification of Peru11 in a cat and BEB6 in a dog constitutes the first report of those genotypes in such hosts as well as first report in Spain. This is also the first evidence of genotype D in cats and PtEb IX in dogs in Spain. Three out of the four genotypes, BEB6, D and Peru11, have been previously reported as human pathogens and are potentially zoonotic indicating that dogs and cats need to be considered potential sources of human infection and environmental contamination.
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http://dx.doi.org/10.1007/s00436-019-06428-1DOI Listing
October 2019

Absence of Neospora caninum DNA in Human Clinical Samples, Spain.

Emerg Infect Dis 2019 06;25(6):1226-1227

Low antibody titers to Neospora caninum have been reported in humans, but infection has not been confirmed. We used N. caninum-specific PCR to test 600 clinical samples from patients with toxoplasmosis signs but Toxoplasma gondii-negative PCR results. We did not detect N. caninum DNA, demonstrating it is an unlikely opportunistic zoonotic agent.
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http://dx.doi.org/10.3201/eid2506.181431DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6537714PMC
June 2019
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