Immunoproteomic and mass spectrometric analysis of Eimeria acervulina antigens recognized by antisera from chickens infected with E. acervulina, E. tenella or E. necatrix

Abstract

Background: Coccidiosis is caused by Eimeria spp. and can result in severe economic losses to the global poultry industry. Due to anticoccidial drug resistance rapidly developing in the parasites and drug residues in poultry products, efficacious and safe alternative coccidia control measures are needed. The objective of the present study was to identify common protective antigens which may be used as vaccine candidates in the development of subunit, multivalent, cross-protective vaccines against most of the economically important Eimeria species.Methods: Whole sporozoite proteins of Eimeria acervulina were prepared and analyzed by 2-dimensional gel electrophoresis (2-DE) followed by western blotting using immune sera specific to E. tenella, E. acervulina, or E. necatrix. The protein spots detected by all three immune sera were then excised from the preparative gel and protein ID was performed by MALDI-TOF-MS/MS. Results:Approximately 620 E. acervulina sporozoite protein spots were demonstrated by 2-DE with silver staining, among which 23 protein spots were recognized by immune sera specific to all three Eimeria species. The results showed that 21 putative E. acervulina proteins were identified, which include proteins with known enzymatic properties, and those which are involved in protein translation, transport and trafficking, and ribosomal biogenesis and functions. There is one protein which may be involved in transcription and one heat-shock protein. Two proteins contain predicted domains, but with no apparent functions known. There were 2 protein spots which had no detectable proteins. None of the proteins has a predicted signal peptide or a transmembrane domain; however, 6 of the 21 putative proteins were predicted to be potentially secretory through the non-classical pathway. Conclusions:Our study identified a diverse group of antigens immunologically common to all three Eimeria species, none of which was previously characterized and tested as a vaccine candidate. Further research on immunogenicity and cross-protective potential of these individual proteins as vaccine candidates will aid the development of vaccines against the most common and pathogenic Eimeria spp.

Immunoproteomic analysis of fish ectoparasite, Argulus siamensis antigens

“…Antiserum-based immunoproteomic studies have been carried out for identifying such potential antigens from several parasites including ectoparasites. 25,26 Initially, the A siamensis parasite species was identified by PCR and maintained in wet laboratory on rohu as a host for our experiments. We developed immune serum in rohu to probe Argulus antigens in Western blot transferred from the 2D gel to identify the immunoreactive proteins.…”

Pathological changes and antigen localization in the small intestine of rabbits infected with Eimeria magna

“…Studies on coccidial antigens have mostly identified a variety of common antigens and proteins, but they have been mostly limited to chicken coccidiosis; only E. stiedai has been previously studied in rabbits (Song et al, 2017). Liu et al (2020) identified twenty-one Eimeria acervulina sporozoite antigens that can be recognised by antisera from chickens infected with E. acervulina, E. tenella or E. necatrix. Studies in chickens and our results support the need for further comparisons of coccidial antigens parasitising rabbits.…”

Vaccines against chicken coccidiosis with particular reference to previous decade: progress, challenges, and opportunities

“…Over many decades, Eimeria vaccines have gained a considerable importance, by virtue of effectively preventing the disease without developing resistance in field strains as in anticoccidial chemotherapeutics (Sander et al 2019 ; Liu et al 2020 ; Zhao et al 2020 ). Rather, the sensitivity towards previously resistant chemical anticoccidials has been enhanced by the use of Eimeria vaccines (Jenkins et al 2010 ).…”