title: RAPD analysis and sequencing of ITS1/5.8S rRNA/ITS2 and Fe-hydrogenase as tools for genetic classification of potentially pathogenic isolates of Trichomonas gallinae.
abstract: Trichomonas gallinae is a worldwide parasite that causes oropharyngeal avian trichomonosis. During eight years, 60 axenic isolates were obtained from different bird species and characterized by three molecular methods: RAPD analysis and PCR-sequencing of ITS1/5.8S rRNA/ITS2 fragment and Fe-hydrogenase gene. We have found two genotypes of ITS1/5.8S rRNA/ITS2 widely distributed among bird populations, a new variant and also two sequences with mixed pattern. Genotype ITS-OBT-Tg-1 was associated with the presence of gross lesions in birds. We have found eight genotypes of the Fe-hydrogenase (A1, A2, C2, C2.1, C4, C5, C6 and C7), three of them are new reports (C5, C6 and C7), and also three sequences with mixed pattern. Subtype A1 of the Fe-hydrogenase was also related with the presence of lesions. RAPD analyses included most of the strains isolated from animals with lesions in one of the sub-clusters. Potentially pathogenic isolates of T. gallinae obtained in this study fulfill the following criteria with one exception: isolated from lesions+ITS-OBT-Tg-1 genotype+FeHyd A1+RAPD sub-cluster I2.
authors.affiliation: Universidad Católica de Valencia, Facultad de Veterinaria y Ciencias Experimentales, Departamento de Producción Animal y Salud Pública, Calle Guillem de Castro, 94, 46003, Valencia, Spain. Electronic address: jose.sansano@ucv.es.
authors.forename: José
authors.initials: J
authors.lastname: Sansano-Maestre
authors.affiliation: Universidad CEU Cardenal Herrera, Instituto de Ciencias Biomédicas, Facultad de Veterinaria, Departamento de Producción y Sanidad Animal, Salud Pública Veterinaria y Ciencia y Tecnología de los Alimentos, Calle Tirant Lo Blanc, 7, 46115 Alfara del Patriarca, Valencia, Spain. Electronic address: maria.martinez1@uchceu.es.
authors.forename: María Del Carmen
authors.initials: MDC
authors.lastname: Martínez-Herrero
authors.affiliation: Universidad CEU Cardenal Herrera, Instituto de Ciencias Biomédicas, Facultad de Veterinaria, Departamento de Producción y Sanidad Animal, Salud Pública Veterinaria y Ciencia y Tecnología de los Alimentos, Calle Tirant Lo Blanc, 7, 46115 Alfara del Patriarca, Valencia, Spain. Electronic address: marilena@uchceu.es.
authors.forename: María Magdalena
authors.initials: MM
authors.lastname: Garijo-Toledo
authors.affiliation: Universidad Complutense de Madrid, Facultad de Veterinaria, Departamento de Sanidad Animal, Avenida Puerta de Hierro s/n, 28040 Madrid, Spain. Electronic address: mariateg@ucm.es.
authors.forename: María Teresa
authors.initials: MT
authors.lastname: Gómez-Muñoz
mesh_terms: D000818:Animals; D001715:Bird Diseases / Q000469:parasitology; D001717:Birds; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D005838:Genotype; D006864:Hydrogenase / Q000235:genetics / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D019105:Random Amplified Polymorphic DNA Technique; D014244:Trichomonas / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D014245:Trichomonas Infections / Q000469:parasitology / Q000662:veterinary
chemical_list: D021903:DNA, Ribosomal Spacer; D007506:Iron-Sulfur Proteins; C118830:iron hydrogenase; D006864:Hydrogenase
keywords: Fe-hydrogenase gene; Genotype; ITS1/5.8S rRNA/ITS2; Pathogenic; RAPD; Trichomonas gallinae