Validation of the TrichinEasy® digestion system for the detection of Anisakidae larvae in fish products

Abstract

Anisakis and other parasites belonging to the Anisakidae family are organisms of interest for human health, because of their high zoonotic potential. Parasites belonging to this family can cause Anisakiasis, a parasitological disease caused by the ingestion of raw, infested fish products. Furthermore, evidence from the EFSA (European Food Safety Authority; EFSA 2010) has highlighted the allergological potential of nematodes belonging to the Anisakis genre. The detection and identification of Anisakidae larvae in fish products requires an initial visual inspection of the fish sample, as well as other techniques such as candling, UV illumination and artificial digestion. The digestion method consists of the simulation of digestive mechanics, which is made possible by the utilization of HCl and pepsin, according to EC Regulation 2075/2005. In this study, a new Anisakidae larvae detection method using a mechanical digestion system called Trichineasy® was developed. A total of 142 fish samples, belonging to 14 different species, were examined to validate the method. A reaction mixture with 100 g of sample, 10 g of pepsin (1:10000 NF) and 50 ml of 10% HCl at 36 ± 1°C for 20 minutes was evaluated to be the best condition for the digestion of fish samples. These parameters have also allowed the detection of viable larvae after digestion. The results confirm this instrumentation as a valuable and safe tool for the detection of Anisakidae larvae in fishery products.

Incidence of Parasites on Frozen Alaska Pollock (Gadus chalcogrammus, Pallas, 1814) Fillets

“…The presence of parasites in fish can be assessed using different methodologies: visual inspection and transillumination, known as candling table (regulated), destructive examination, pressing and observation with ultraviolet light [18], spectrophotometry [19], artificial digestion [20] and molecular techniques (PCR-RFLP, ITS-1 region) [21]. Studies such as the one by Fraulo and coauthors [22] and Cammilleri and coauthors [20] demonstrate that the candling table technique is not the most suitable for the detection of parasites in fish, which digestion techniques followed by identification of molecules are high sensitivity. But this technique Brazilian Archives of Biology and Technology.…”

Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products

“…All the processed fish samples came from Italian supermarkets. The Anisakidae larvae used for the artificial contamination of the samples were collected from Lepidopus caudatus, Clupea harengus, and Merluccius merluccius samples after visual inspection and modified chloro-peptic digestion [40]. The larvae isolated were washed in physiological saline serum (pH 7) and morphologically identified by B-800 light microscopy (Optika, Ponteranica, Italy) according to taxonomic keys [41].…”

Immune response of teleost fish to helminth parasite infection

“…As an alternative, molecular techniques as real-time polymerase chain reaction (PCR) with high-resolution molecular analysis was employed to detect and quantify the presence of Anisakis simplex in fish, enabling even taxa differentiation of other parasites (JAISWAL et al, 2017). For fish products, usually it is used UV illumination, artificial digestion by the utilization of chloridric acid and pepsin, and recently it was developed a new method called TrichinEasy  digestion system, a confirmatory test of the presence of the larvae (CAMMILLERI et al, 2016).…”