Keberadaan Anisakis typica (Anisakidae) dari Ikan Tongkol dan Ikan Layang dari perairan Sulawesi Barat

Muhammad Dusil Hafid(1*), Hilal Anshary(2)

(1) Fish Quarantine and Inspection Agency, Fish quarantine station at West Sulawesi, Mamuju, Indonesia
(2) Laboratory of Fish Parasites and Diseases, Faculty of Marine Science and Fisheries, Hasanuddin University, Indonesia
(*) Corresponding Author


Anisakis (L3-stage) is a parasitic nematode commonly found in marine fish or squid serve as intermediate or paratenic host. The purpose of this research was to identify Anisakis larvae using PCR-RFLP and sequencing methods. A total of 30 individuals of each species: bullet tuna (Auxis rochei) and Indian scad
(Decapterus russeli) were examined for presence of Anisakis spp. Fish samples caught by fishermen around Mamuju waters were purchased at a fish market. The fish was necropsied and the organs: liver, intestine wall, heart and muscle were removed and put on separated petri dishes and examined for parasites under a stereo microscope. Anisakis found was cleaned and fixed in 70% alcohol. Initial identification was based on ventriculus shape and presence of mucron to distinguish Anisakis type I and Anisakis type II. All Anisakis found belong to Anisakis type I. Ten Anisakis type I were isolated, cleaned and stored in microfuge tubes containing 70% alcohol. The parasite gDNA was extracted using Wizard genomic DNA extraction and purification kit (Promega).
Species of Anisakis was determined by PCR-RFLP at ITS1-5.8S-ITS2 region using two restriction enzymes: Taq I and Hinf I and PCR-sequencing in mtDNA cox2 region. Analysis by PCR-RFLP showed that all Anisakis type I
examined was in the same pattern as Anisakis typica. Sequencing in mtDNAcox-2 region and the phylogenetic analysis showed that all samples were in the same cluster as A. typica. Based on PCR-RFLP and sequencing
analysis, all Anisakis found in this study belong to A. typica.


Anisakis; marine fish; mtDNA cox-2; PCR-RFLP; Sulawesi

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