Quantitative Proteomics Comparison of Total Expressed Proteomes of Anisakis simplex Sensu Stricto, A. pegreffii, and Their Hybrid Genotype

Abstract

The total proteomes of Anisakis simplex s.s., A. pegreffii and their hybrid genotype have been compared by quantitative proteomics (iTRAQ approach), which considers the level of expressed proteins. Comparison was made by means of two independent experiments considering four biological replicates of A. simplex and two each for A. pegreffii and hybrid between both species. A total of 1811 and 1976 proteins have been respectively identified in the experiments using public databases. One hundred ninety-six proteins were found significantly differentially expressed, and their relationships with the nematodes’ biological replicates were estimated by a multidimensional statistical approach. Results of pairwise Log2 ratio comparisons among them were statistically treated and supported in order to convert them into discrete character states. Principal component analysis (PCA) confirms the validity of the method. This comparison selected thirty seven proteins as discriminant taxonomic biomarkers among A. simplex, A. pegreffii and their hybrid genotype; 19 of these biomarkers, encoded by ten loci, are specific allergens of Anisakis (Ani s7, Ani s8, Ani s12, and Ani s14) and other (Ancylostoma secreted) is a common nematodes venom allergen. The rest of the markers comprise four unknown or non-characterized proteins; five different proteins (leucine) related to innate immunity, four proteolytic proteins (metalloendopeptidases), a lipase, a mitochondrial translocase protein, a neurotransmitter, a thyroxine transporter, and a structural collagen protein. The proposed methodology (proteomics and statistical) solidly characterize a set of proteins that are susceptible to take advantage of the new targeted proteomics.

Proteomic Profiling and In Silico Characterization of the Secretome of Anisakis simplex Sensu Stricto L3 Larvae

“…This study is the first global proteomic analysis of the A. simplex (s.s.) L3 larval secretome. Previous proteomic investigations of Anisakis did not include profiling of the secretome proteins [13,[31][32][33][34][35][36][37][38]. Therefore, knowledge on A. simplex ES proteins is very fragmented, although important aspects related to metabolism, pathogenicity, and hostparasite interactions are known to be associated with ES proteins.…”

Re‐evaluation of certain aspects of the EFSA Scientific Opinion of April 2010 on risk assessment of parasites in fishery products, based on new scientific data. Part 1: ToRs1–3

“…The first profiling of A. simplex (s. s.) and A. pegreffii was done using the MALDI‐TOF/TOF MS approach (Arcos et al., 2014 ). This was followed by mapping of allergens in the Anisakis proteome using the nano LC‐ESI‐MS/MS technique (Fæste et al., 2014 ), quantification of A. simplex proteins in food matrix by LC‐MS/MS (Fæste et al., 2016 ), fast monitoring of parasites in fish products using parallel reaction monitoring (PRM) MS (Carrera et al., 2016 ) and quantification of proteomes of A. simplex s.s., A. pegreffii and their hybrids using iTRAQ labelling and subsequent nano LC‐ESI‐MS/MS (Arcos et al., 2020 ). Shotgun sequence identification of L3 and L4 proteins based on isobaric mass labelling (Tandem Mass Tag, TMT) (Stryiński et al., 2019 , 2022 ) was then undertaken, followed by the construction of a spectral library of Anisakis spp.…”

Novel Omics Studies on Anisakid Nematodes

“…This investigation revealed the presence of specific Anisakis antigens as well as secreted molecules (among them: peptidases, lipase-like and chitinases) shared with phylogenetically related parasitic nematodes. The second proteomics research article by Arcos and collaborators aimed to compare proteomes obtained from different members of A. simplex sensu lato complex (A. simplex s.s., A. pegreffii and their hybrid) [7]. This comparison revealed thirty-seven proteins as discriminant taxonomic biomarkers, most of which exhibited crucial biological functions related to allergy, innate immunity and interactions with the host.…”