Proteomic and Bioinformatic Investigations of Heat-Treated Anisakis simplex Third-Stage Larvae

Kochanowski, Maciej; Różycki, Mirosław; Dąbrowska, Joanna; Bełcik, Aneta; Karamon, Jacek; Sroka, Jacek; Cencek, Tomasz

doi:10.3390/biom10071066

Cited by 13 publications

(11 citation statements)

References 96 publications

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“…However, SDS-PAGE and WB band profiles of Anisakis antigen after heating were clearly visible. Similar observations regarding the reduction of the number and intensity of bands in electrophoretic and WB profiles of autoclaved Anisakis antigen was reported in a previous study [ 34 ]. In the present investigation, we found that the heated antigen retained the ability to bind IgG antibodies and, therefore, it was possible to use them in CL-ELISA tests.…”

Section: Discussionsupporting

confidence: 90%

Development and Application of Novel Chemiluminescence Immunoassays for Highly Sensitive Detection of Anisakis simplex Proteins in Thermally Processed Seafood

et al. 2020

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The third-stage larvae (L3) of Anisakis simplex are the most important source of hidden allergens in seafood products. However, there exist no commercial methods for detecting Anisakis proteins in food. Furthermore, only a few methods have been validated for the detection of A. simplex in thermally processed food. The aims of our study are (i) the development and validation of high-sensitivity chemiluminescent (CL) immunoassays for the detection of A. simplex proteins in processed seafood, (ii) and A. simplex antigen detection in common seafood products from Polish markets. We developed and validated CL sandwich ELISA (S-ELISA) and CL competitive ELISA (C-ELISA) methods for A. simplex proteins detection in food, with respective detection limits of 0.5 and 5 ng/mL. The usefulness of the assays for detecting A. simplex proteins in highly processed food was evaluated by examination of autoclaved canned fish spiked with A. simplex larvae (1–8 larvae/200 g). Commercial real-time PCR was unable to detect A. simplex in autoclaved samples at all levels of enrichment with Anisakis larvae. CL-S-ELISA was used to test various types of seafood products from Polish markets. Among all tested products (n = 259), 28% were positive. A. simplex antigens were found mostly (n = 39) in smoked fish products: mackerel, herring, cod, and hake. Other positive samples were found in marinated herrings, canned cod livers, canned mackerels, and surimi sticks. In tuna, Atlantic argentine, anchovy, sardine, sprat, and squid products, A. simplex antigens were not detected. This study provides novel effective tools for the detection of A. simplex proteins in processed food and highlights the potential allergic hazards for Anisakis-sensitized Polish consumers of seafood.

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Section: Discussionsupporting

confidence: 90%

Development and Application of Novel Chemiluminescence Immunoassays for Highly Sensitive Detection of Anisakis simplex Proteins in Thermally Processed Seafood

et al. 2020

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“…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.…”

Section: Discussionmentioning

confidence: 99%

“…Of the seven A. simplex ES allergens listed by WHO/IUIS [89], only Ani s 4 was detected in the present study. Ani s 4 is significant because of its heat-and pepsin-resistant properties and its ability to cause anaphylaxis [13,90,91]. Other Anisakis allergens were presumably not expressed in the in vitro culture conditions, or their concentration was below the limit of detection of LC-MS/MS.…”

Section: Discussionmentioning

confidence: 99%

“…Another very important form of the disease is gastroallergic anisakiasis, in which abdominal symptoms are accompanied by allergic-like reactions such as urticaria, angioedema, or even anaphylaxis [1,11]. Furthermore, studies have suggested that dead Anisakis larvae can cause allergic reactions in sensitized humans [12][13][14]. From an anatomopathological point of view, Anisakis larva invasion leads to the development of inflammation (predominantly eosinophilic), edema, hemorrhage, formation of granuloma, and ulcers at the larval location, which is usually in the mucosa and submucosa of the gastrointestinal wall [10,12].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2022

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Anisakis simplex sensu stricto (s.s.) L3 larvae are one of the major etiological factors of human anisakiasis, which is one of the most important foodborne parasitic diseases. Nevertheless, to date, Anisakis secretome proteins, with important functions in nematode pathogenicity and host-parasite interactions, have not been extensively explored. Therefore, the aim of this study was to identify and characterize the excretory-secretory (ES) proteins of A. simplex L3 larvae. ES proteins of A. simplex were subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, and the identified proteins were then analyzed using bioinformatics tools. A total of 158 proteins were detected. Detailed bioinformatic characterization of ES proteins was performed, including Gene Ontology (GO) analysis, identification of enzymes, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis, protein family classification, secretory pathway prediction, and detection of essential proteins. Furthermore, of all detected ES proteins, 1 was identified as an allergen, which was Ani s 4, and 18 were potential allergens, most of which were homologs of nematode and arthropod allergens. Nine potential pathogenicity-related proteins were predicted, which were predominantly homologs of chaperones. In addition, predicted host-parasite interactions between the Anisakis ES proteins and both human and fish proteins were identified. In conclusion, this study represents the first global analysis of Anisakis ES proteins. The findings provide a better understanding of survival and invasion strategies of A. simplex L3 larvae.

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“…Finally, the peptides were filtered at e-value < 0.05 for the false discovery rate (FDR) using the Mascot searching engine; that is to say, the ion scores were higher than corresponding identity scores 57,58 . The manual method and the PepEx (https://github.com/ eparker05/Pepti-deExtractor) were both used to map the distribution of peptides onto their full-length conopeptide protein sequences.…”

Section: Peptide Identificationmentioning

confidence: 99%

The first Conus genome assembly reveals a primary genetic central dogma of conopeptides in C. betulinus

et al. 2021

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Although there are various Conus species with publicly available transcriptome and proteome data, no genome assembly has been reported yet. Here, using Chinese tubular cone snail (C. betulinus) as a representative, we sequenced and assembled the first Conus genome with original identification of 133 genome-widely distributed conopeptide genes. After integration of our genomics, transcriptomics, and peptidomics data in the same species, we established a primary genetic central dogma of diverse conopeptides, assuming a rough number ratio of ~1:1:1:10s for the total genes: transcripts: proteins: post-translationally modified peptides. This ratio may be special for this worm-hunting Conus species, due to the high diversity of various Conus genomes and the big number ranges of conopeptide genes, transcripts, and peptides in previous reports of diverse Conus species. Only a fraction (45.9%) of the identified conotopeptide genes from our achieved genome assembly are transcribed with transcriptomic evidence, and few genes individually correspond to multiple transcripts possibly due to intraspecies or mutation-based variances. Variable peptide processing at the proteomic level, generating a big diversity of venom conopeptides with alternative cleavage sites, post-translational modifications, and N-/C-terminal truncations, may explain how the 133 genes and ~123 transcripts can generate thousands of conopeptides in the venom of individual C. betulinus. We also predicted many conopeptides with high stereostructural similarities to the putative analgesic ω-MVIIA, addiction therapy AuIB and insecticide ImI, suggesting that our current genome assembly for C. betulinus is a valuable genetic resource for high-throughput prediction and development of potential pharmaceuticals.

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Proteomic and Bioinformatic Investigations of Heat-Treated Anisakis simplex Third-Stage Larvae

Cited by 13 publications

References 96 publications

Development and Application of Novel Chemiluminescence Immunoassays for Highly Sensitive Detection of Anisakis simplex Proteins in Thermally Processed Seafood

Development and Application of Novel Chemiluminescence Immunoassays for Highly Sensitive Detection of Anisakis simplex Proteins in Thermally Processed Seafood

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

The first Conus genome assembly reveals a primary genetic central dogma of conopeptides in C. betulinus

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