Comparative Proteomics Analysis of Anisakis simplex s.s.-Evaluation of the Response of Invasive Larvae to Ivermectin.

Iwona Polak,Robert Stryiński,Elżbieta Łopieńska-Biernat,Jesús Mateos,Mónica Carrera

doi:10.3390/genes11060710

Iwona Polak, Robert Stryiński + Show 3 more

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https://doi.org/10.3390/genes11060710

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Abstract

Ivermectin (IVM), an antiparasitic drug, has a positive effect against Anisakis simplex s.s. infection and has been used for the treatment and prevention of anisakiasis in humans. However, the molecular mechanism of action of IVM on A. simplex s.s. remains unknown. Herein, tandem mass tag (TMT) labeling and extensive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis were used to identify the effect of IVM on the proteome of A. simplex s.s. in vitro. During the study, 3433 proteins, of which 1247 had at least two protein unique peptides, were identified. Comparative proteomics analysis revealed that 59 proteins were differentially regulated (DRPs) in IVM-treated larvae, of which 14 proteins were upregulated and 38 were downregulated after 12 h of culture, but after 24 h, 12 proteins were upregulated and 22 were downregulated. The transcription level of five randomly selected DRPs was determined by real-time PCR as a supplement to the proteomic data. The functional enrichment analysis showed that most of the DRPs were involved in oxidoreductase activity, immunogenicity, protein degradation, and other biological processes. This study has, for the first time, provided comprehensive proteomics data on A. simplex s.s. response to IVM and might deliver new insight into the molecular mechanism by which IVM acts on invasive larvae of A. simplex s.s.

Highlights

Anisakiasis is a zoonotic disease triggered by the third-stage larvae of nematodes from the genusAnisakis
The transcription level of five randomly selected differentially regulated proteins (DRPs) was determined by real-time PCR as a supplement to the proteomic data
The metabolic processes activated in parasites during the host drug treatment are still not fully understood

Summary

Introduction

Anisakiasis is a zoonotic disease triggered by the third-stage larvae of nematodes from the genusAnisakis. Anisakiasis is a zoonotic disease triggered by the third-stage larvae of nematodes from the genus. Intermediate and/or paratenic hosts of the larvae are crustaceans, cephalopods, and fish [1]. Infection occurs upon consumption of raw or undercooked marine fish and crustaceans contaminated with the third-stage larvae of A. simplex [2]. Infected dishes of raw fish, such as sushi and sashimi, as well as the consumption of marinated or raw fish in European countries such as Italy and Spain, are significant sources of the infection [3]. Greater awareness of Anisakis infection has resulted in an increase in the frequency of reported anisakiasis cases in many more countries, besides the expected ones (Italy, Spain, Japan), e.g., in Poland the first case of this disease was described in 2020 [4,5,6,7,8]

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