On the morphology of Anisakis pegreffii: a comparative analysis of threeudmicroscopic techniques used to build a new parasite atlas

摘要

BACKGROUND: Human anisakidosis is a parasitic anthropozoonosis caused by larvaludnematodes of the family Anisakidae. Here, we report a detailed description of theudmorphology of Anisakis pegreffii third-stage larva performed using a conventional lightudand confocal microscopy, and scanning electron microscopy (SEM) that provide audbasis for both phenotypic studies and genetic mutations.udMETHODS: The collected larvae from fish were morphologically identified as Anisakisudlarvae Type I, and they were characterized by PCR-RFLP to identify the Anisakisudpegreffii specie. Using NC5/NC2 primers, ribosomal genomic regions ITS1, 5.8 SrRNAudand ITS2 of DNA were amplified and PCR products were sequenced. Fifteen larvaeudbelonging to Anisakis pegreffii were fixed, sectioned, and examined with a light andudconfocal microscope and by SEM.udRESULTS: In our studies, have been acquired detailed ultrastructural images, whichudhave been integrated with those derived from the dissection of the parasite, obtainedudwith light and confocal microscopy. The structural and ultrastructural imagesudconcerning the third stage larvae of Anisakis pegreffii have been studied, analyzed andudcompared among them. The derived overall view has allowed detecting new interestinguddetails of a well-known parasite and has been schematically showed.udCONCLUSIONS: The aim of this study is to furnish an updated atlas of Anisakisudpegreffii. Confocal microscopy, as well as the light and electron microscopy haveudplayed a pivotal role in the accumulation of new scientific data regarding the anatomicaludstructures of this nematode. This work is the result of one year of engagement by theudAuthors and the outcome is a comprehensive atlas on Anisakis pegreffii microscopy.