Person: Fernández Escobar, Mercedes
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First Name
Mercedes
Last Name
Fernández Escobar
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Sanidad Animal
Area
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Publication Toxoplasma gondii Genetic Diversity in Mediterranean Dolphins(MPDI, 2022-08-12) Fernández Escobar, Mercedes; Giorda, Federica; Mattioda, Virgina; Audino, Tania; Di Nocera, Fabio; Lucifora, Giuseppe; Varello, Katia; Grattarola, Carla; Ortega Mora, Luis Miguel; Casalone, Cristina; Calero Bernal, RafaelToxoplasma gondii constitutes a major zoonotic agent but also has been frequently identified as an important cause of clinical disease (e.g., abortion, pneumonia, encephalitis) in wildlife; specifically, T. gondii has been associated with neurological disease in cetaceans. This study investigated the genetic diversity of T. gondii strains involved in infections in dolphins found stranded in the Mediterranean coastlines of Italy. Tissue samples from 16 dolphins (Stenella coeruleoalba and Tursiops truncatus species) positive for T. gondii-DNA presence by PCR were examined by histology and subjected to further genetic characterization of strains detected by PCR-RFLP and multilocus PCR-sequencing assays. According to fully genotyped samples, the genotypes ToxoDB#3 (67%) and #2 (22%) were detected, the latter being reported for the first time in cetaceans, along with a mixed infection (11%). Subtyping by PCR-seq procedures provided evidence of common point mutations in strains from southwestern Europe. Despite evidence of T. gondii as a cause of neurological disease in dolphins, sources of infections are difficult to identify since they are long-living animals and some species have vast migration areas with multiple chances of infection. Finally, the genetic diversity of T. gondii found in the dolphins studied in the Mediterranean coastlines of Italy reflects the main genotypes circulating inland in the European continent.Publication Effects of Ovine Monocyte-Derived Macrophage Infection by Recently Isolated Toxoplasma gondii Strains Showing Different Phenotypic Traits(MPDI, 2022-12-07) Vallejo, Raquel; Benavides, Julio; Arteche Villasol, Noive; Fernández Escobar, Mercedes; Ferreras, María Del Carmen; Pérez, Valentín; Gutierrez Expósito, DanielOvine toxoplasmosis is one the most relevant reproductive diseases in sheep. The genetic variability among different Toxoplasma gondii isolates is known to be related to different degrees of virulence in mice and humans, but little is known regarding its potential effects in sheep. The aim of this study was to investigate the effect of genetic variability (types II (ToxoDB #1 and #3) and III (#2)) of six recently isolated strains that showed different phenotypic traits both in a normalized mouse model and in ovine trophoblasts, in ovine monocyte-derived macrophages and the subsequent transcript expression of cytokines and iNOS (inducible nitric oxide synthase). The type III isolate (TgShSp24) showed the highest rate of internalization, followed by the type II clonal isolate (TgShSp2), while the type II PRU isolates (TgShSp1, TgShSp3, TgShSp11 and TgShSp16) showed the lowest rates. The type II PRU strains, isolated from abortions, exhibited higher levels of anti-inflammatory cytokines and iNOS than those obtained from the myocardium of chronically infected sheep (type II PRU strains and type III), which had higher levels of pro-inflammatory cytokines. The present results show the existence of significant intra- and inter-genotypic differences in the parasite-macrophage relationship that need to be confirmed in in vivo experiments.Publication Evidence for Unknown Sarcocystis-Like Infection in Stranded Striped Dolphins (Stenella coeruleoalba) from the Ligurian Sea, Italy(MPDI, 2021-04-22) Giorda, Federica; Romani Cremaschi, Umberto; Marsh, Antoinette E.; Grattarola, Carla; Iulini, Barbara; Pautasso, Alessandra; Varello, Katia; Berio, Enrica; Gazzuola, Paola; Marsili, Letizia; Di Francesco, Cristina E.; Goria, Maria; Verna, Federica; Audino, Tania; Peletto, Simone; Caramelli, Maria; Fernández Escobar, Mercedes; Sierra, Eva; Fernández, Antonio; Calero Bernal, Rafael; Casalone, CristinaTwo 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 Toxoplasma gondii, additional histopathological, immunohistochemical, ultrastructural, and biomolecular investigations were performed, aiming to fully characterize the organism. Histopathology revealed the presence of large Sarcocystis-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 Sarcocystis neurona, Sarcocystis falcatula, Hammondia hammondi, and Neospora caninum. T. gondii coinfection was confirmed only in dolphin SD2. Sarcocystis 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 Sarcocystis-like tissue cysts in the brain of stranded cetaceans along with the first description of Sarcocystis sp. infection in muscle tissue of dolphins from the Mediterranean basin.Publication Genetic and phenotypic characterization of Toxoplasma gondii isolates obtained from sheep and Iberian pigs in Spain(Universidad Complutense de Madrid, 2022-02-17) Fernández Escobar, Mercedes; Ortega Mora, Luis Miguel; Collantes Fernández, Esther; Calero Bernal, RafaelToxoplasma gondii is an apicomplexan parasite globally distributed with a heteroxenous life cycle that virtually comprises all homoeothermic animals, including humans, as intermediate hosts and felids as definitive hosts. The zoonotic, abortifacient, and foodborne nature of the parasite makes toxoplasmosis a relevant public and animal health concern worldwide.A comprehensive research effort on T. gondii biology along with the rapid development of molecular techniques suitable for strains genotyping over the last decades, led to the initial description of a widely clonal European and North American T. gondii genetic population dominated by three main clonal genetic types (I, II, and III), in contrast to an extremely diverse South American population. However, the information available from Europe is limited, with frequent methodological deficiencies and important sampling disparities among regions. Briefly, the available European literature evidences a clear predominance of type II strains (comprising around 80% of samples) coexisting with much less abundant type III and recombinant strains or mixed infections, as well as minor proportions of type I and imported genotypes. In the specific case of Spain, the majority of the scarce investigations dealt with direct genotyping from clinical samples, with the subsequent limitations to classify the strains and the impossibility of extending its characterization...Publication Dynamics of Neospora caninum-Associated Abortions in a Dairy Sheep Flock and Results of a Test-and-Cull Control Programme(MPDI, 2021-11-20) Sánchez Sánchez, Roberto; Vázquez Calvo, Ángela; Fernández Escobar, Mercedes; Regidor Cerrillo, Javier; Benavides, Julio; Gutiérrez, Jorge; Gutierrez Expósito, Daniel; Crespo Ramos, Francisco José; Ortega Mora, Luis Miguel; Álvarez García, GemaNeospora caninum is an apicomplexan parasite that can cause abortions and perinatal mortality in sheep. Although ovine neosporosis has been described worldwide, there is a lack of information about the relationship between N. caninum serostatus and the reproductive performance. In this study, we described the infection dynamics in a dairy sheep flock with an abortion rate up to 25% and a N. caninum seroprevalence of 32%. Abortions were recorded in 36% and 9% of seropositive and seronegative sheep, respectively. Seropositive sheep were more likely to abort twice (OR = 4.44) or three or more times (OR = 10.13) than seronegative sheep. Endogenous transplacental transmission was the main route of transmission since 86% of seropositive sheep had seropositive offspring. Within dams that had any abortion, seropositive sheep were more likely than seronegative ones to have female descendants that aborted (OR = 8.12). The slight increase in seropositivity with the age, the low percentage of animals with postnatal seroconversion or with low avidity antibodies, and the seropositivity of one flock dog, indicated that horizontal transmission might have some relevance in this flock. A control programme based on selective culling of seropositive sheep and replacement with seronegative animals was effective in reducing the abortion rate to 7.2%.