CONICET | Buscador de Institutos y Recursos Humanos

Coccidiosis caused by Eimeria spp. has an important economic significance for the poultry industry. E. tenella (Et) and E. acervulina (Ea) are 2 of the 3 most frequent species, the former associated with a high rate of mortality. Although there are many effective live vaccines available in the market, they have some disadvantages that promote the study of novel antigens and the development of new vaccines based on alternative technologies. Given that glycosylphosphatidylinositol (GPI)-anchored proteins, displayed on the surface of the parasite, are good vaccine candidates, the objective of this work was their selection in Et and Ea through reverse vaccinology and the study of polymorphisms from new native isolates. A sequential pipeline, previously applied to related Apicomplexa, was followed and 3 Et and 3 Ea genes for polymorphism analysis, based on i. Presence of GPI anchor and absence of transmembrane sites; ii. Surface localization; iii. Evidence of sporozoite transcription, and iii. No of strong and weak binders in MHC I predicted sites. Specific oligonucleotides were designed for PCRs from Eimeria-DNA of Argentinian isolates (n=4). Amplicons were subjected to NGT sequencing, and sequence multiple alignments were done upon identity confirmation. Fifteen Ea and 89 Et GPI-proteins were predicted from the proteomes and secretomes (6,873 and 428 Ea proteins, and 8,627 and 624 Et proteins, respectively) and a short list of 8 Ea or 22 Et proteins was produced from which etsag1, etsag, ethyp, easag4, easag2 and eahyp were selected for sequencing. ethyp, easag2, and eahyp have demonstrated 100% of conservation among isolates and the reference sequence, etsag1 from 1 isolate showed 99.91 %, containing 2 mutations; etsag from 3 isolates showed 99.88 % with 2 mutations and easag2 from 1 isolate showed 99.66 % with 4 mutations. In conclusion through a bioinformatic approach, conserved potential vaccine antigens could be selected

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