Characterisation of common hypothetical surface peptides between protozoan parasites (Perkinsus olseni) originating from different geographical locations.

Perkinsus olseni and P. marinus are classified as notifiable pathogens by the World Organisation for Animal Health and are known to cause perkinsosis in a variety of molluscs globally. Mass mortalities due to these parasites in farms and in the wild have been a recurrent issue. Diagnosis for these protozoans is currently done using Ray's fluid thioglycollate medium method followed by optical microscopy or molecular assays. Both require a high level of skill and are time-consuming. An immunoassay method would make the diagnosis of perkinsosis quicker and cheaper. The present study used mass spectrometry-based proteomics to investigate common hypothetical surface peptides between different geographical isolates of P. olseni, which could be used to develop immunoassays in the future. Two peptides were identified: POLS_08089, which is a 42.7 kDa peptide corresponding to the 60S ribosomal subunit protein L4; and POLS_15916, which is a conserved hypothetical protein of 55.6 kDa. The identification of peptides may allow the development of immunoassays through a more targeted approach.

Immunoassays and diagnostic antibodies for Perkinsus spp. pathogens of marine molluscs.

Perkinsus sp. protozoans are parasites of a wide variety of molluscs around the world and are responsible for episodes of mass mortalities and large economic losses for aquaculture industries and fisheries. The first step towards the management of infectious episodes is the reliable detection of Perkinsus species. While historic methods for diagnosis of Perkinsus sp. infections in mollusc hosts include histological, in vitro, molecular-genetic, and immunoassays, antibody-based diagnostic assays may prove most practical with development of improved reagents and techniques. This paper reviews historic developments of antibodies against Perkinsus species, and of diagnostic immunoassays. Thirteen research papers reported the development of antibodies against Perkinsus sp. or their extracellular products, mainly P. olseni and P. marinus. Nine of those tested the cross-reactivity of their antibodies against different life stages or species than the one used as immunogen. While all antibodies raised against trophozoites labelled hypnospores, several antibodies raised against hypnospores did not label trophozoites, suggesting antigenic differences between those cell types. Antibody specificity studies showed that there is antigenic heterogeneity between Perkinsus species and Perkinsus-like organisms, and also that common epitopes occur among Perkinsus species, as well as some dinoflagellates. This review summarizes the current knowledge and aims at helping the future development of Perkinsus species-specific antibodies and immunoassays.