Project description:The parasite species complex Anisakis simplex sensu lato (Anisakis simplex sensu stricto; (A. simplex s.s.), A. pegreffii, A. simplex C) is the main cause of severe anisakiasis (allergy) worldwide and is now an important health matter. In this study, the relationship of this Anisakis species complex and their allergenic capacities is assessed by studying the differences between the two most frequent species (A. simplex s.s., A. pegreffii) and their hybrid haplotype by studying active L3 larvae parasiting Merluccius merluccius.
Project description:Protein extracts from Anisakis simplex L3 larvae were autoclaved at 121C for 60 min. Subsequently, prepared samples were analyzed in LC-MS/MS to identify autoclave-resistant proteins (also allergens).
Project description: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. A total of 1,976 proteins have been identified using public databases. One hundred ninety six proteins were found significantly differentially expressed; results of pairwise Log2 ratio comparisons among them were statistically treated and supported in order to convert them into discrete character states. This comparison selected thirty six proteins as discriminant biomarkers among A. simplex, A. pegreffii and their hybrid genotype; eighteen of these biomarkers, encoded by nine 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.
Project description:<p><em>Anisakis simplex</em> are parasitic nematodes that cause anisakiasis. The possibility of infection with this parasite is through consumption of raw or undercooked fish products. <em>A. simplex</em> infections are often misdiagnosed, especially in subclinical cases that do not present with typical symptoms such as urticaria, angioedema and gastrointestinal allergy. The resulting allergic reactions range from rapid-onset and potentially fatal anaphylactic reactions to chronic, debilitating conditions. While there have been numerous published studies on the genomes and proteomes of <em>A. simplex</em>, less attention has been paid to the metabolomes. Metabolomics is concerned with the composition of metabolites in biological systems. Dynamic responses to endogenous and exogenous stimuli are particularly well suited for the study of holistic metabolic responses. In addition, metabolomics can be used to determine metabolic activity at different stages of development or during growth. In this study, we reveal for the first time the metabolomes of infectious stages (L3 and L4) of <em>A. simplex</em> using untargeted metabolomics by ultra-performance liquid chromatography-mass spectrometry. In the negative ionization mode (ESI-), we identified 172 different compounds, whereas in the positive ionization mode (ESI+), 186 metabolites were found. Statistical analysis showed that 60 metabolites were found in the ESI- mode with different concentration in each group, of which 21 were more enriched in the L3 larvae and 39 in the L4 stage of <em>A. simplex</em>. Comparison of the individual developmental stages in the ESI+ mode also revealed a total of 60 differential metabolites, but 32 metabolites were more enriched in the L3 stage larvae, and 28 metabolites were more concentrated in the L4 stage. The metabolomics study revealed that the developmental stages of <em>A. simplex</em> differed in a number of metabolic pathways, including nicotinate and nicotinamide metabolism. In addition, molecules responsible for successful migration within their host, such as pyridoxine and prostaglandins (E1, E2, F1a) were present in the L4 stage. In contrast, metabolic pathways for amino acids, starch and sucrose were mainly activated in the L3 stage. Our results provide new insights into the comparative metabolome profiles of two different developmental stages of <em>A. simplex</em>.</p>
