Assessment of a Sampling Plan Based on Visual Inspection for the Detection of Anisakid Larvae in Fresh Anchovies (Engraulis encrasicolus). A First Step Towards Official Validation?

Guardone, Lisa; Malandra, Renato; Costanzo, Francesco; Castigliego, Lorenzo; Tinacci, Lara; Gianfaldoni, Daniela; Guidi, Alessandra; Armani, Andrea

doi:10.1007/s12161-015-0316-2

Cited by 18 publications

(20 citation statements)

References 39 publications

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“…A subset of Anisakis larvae (from 1 to 4 larvae per product) was submitted to molecular identification. Total DNA extraction was performed according to the protocol used in Guardone et al, (2016b). DNA concentration and purity were determined by a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA).…”

Section: Molecular Identificationmentioning

confidence: 99%

“…The MA threshold was calculated by applying an approach regional law widely used throughout Italy (D'Amico et al, 2014) which defines the maximum number of tolerated larvae in fresh batches of anchovies (three 3 larvae in 10% of the sampled fish). Considering that in the case of fish species caught in large batches, such as anchovies, the number of subjects to collect for a significant sampling is, at least, 29, the maximum number of parasites tolerated is and therefore the MA threshold is 0.3 (Guardone et al, 2016b, Guardone et al, 2017.…”

Section: Mean Abundance (Ma)mentioning

confidence: 99%

“…Moreover, the artificial digestion may also be applied to isolate larvae from fish and it is considered the gold standard for its higher sensitivity (Guardone et al, 2016b;Llarena-Reino et al, 2013). The cuticle of parasitic nematodes has been reported as highly resistant to strong acids and digestive enzymes, regardless of whether the nematodes are live or have been killed by freezing or conventional heating (Tejada et al, 2006).…”

Section: Parasitological Analysismentioning

confidence: 99%

“…On the contrary, they are evident when they infect in high number the celomatic cavity of fish species. This is particularly true in case of fresh fish containing live larvae that which can actively move and become evident also on the external surface (Guardone et al, 2016b). However, dead visible larvae can also be considered a defect according to the definition of the Codex Alimentarius:…”

Section: Dead Anisakid Larvae In Semi-preserved Anchovies: a Potentiamentioning

confidence: 99%

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Anisakis spp. larvae in different kinds of ready to eat products made of anchovies ( Engraulis encrasicolus ) sold in Italian supermarkets

Guardone

Nucera

Lodola

et al. 2018

International Journal of Food Microbiology

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In this study the occurrence of visible anisakid larvae in semi-preserved anchovy products sold on the Italian market was investigated. Totally, 107 ready to eat products (33 salted-ripened, 49 in oil and 25 marinated) were sampled. Each sample was digested, then the digested material was observed under natural and UV light. Parasites were counted, collected and microscopically identified to genus level. A representative subset was molecularly identified using the cox2 gene. At least one visible Anisakis sp. larva was found in 54.2% of the total 107 products analysed and totally 1283 dead larvae were collected. Anisakis sp. larvae were found in all the 33 salted products and 1139 (88.8%) larvae were collected, with a range of 1-105 parasites per product. Larval density per gram was 0.13. Anisakis sp. larvae were found in 49.0% of the products in oil and 143 (11.1%) larvae were isolated, with a range of 0-28 and a density of 0.03. Only 1 larva was found in the 25 marinated products (4.0%, density 0.00). A highly significant difference between all the product categories in respect of number of larvae per product, frequency of products contaminated by at least one larva and larval density per gram was found. Within the subset of larvae molecularly analysed (n=122), 92 (75.4%) were identified as A. pegreffii and 30 (24.6%) as A. simplex. This study showed that semi-preserved anchovy products heavily contaminated with Anisakis spp. larvae reach the market. Beyond the negligible risk for anisakidosis, the presence of dead visible parasites may cause immediate rejection in consumers. In addition, the potential risk related to allergic reactions in sensitized individuals needs to be further assessed. In order to avoid commercialization of obviously contaminated products, fresh anchovies' batches intended for the production of such products should be accurately selected by the processing industry applying inspection methods.

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Section: Molecular Identificationmentioning

confidence: 99%

Section: Mean Abundance (Ma)mentioning

confidence: 99%

Section: Parasitological Analysismentioning

confidence: 99%

Section: Dead Anisakid Larvae In Semi-preserved Anchovies: a Potentiamentioning

confidence: 99%

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Anisakis spp. larvae in different kinds of ready to eat products made of anchovies ( Engraulis encrasicolus ) sold in Italian supermarkets

Guardone

Nucera

Lodola

et al. 2018

International Journal of Food Microbiology

Self Cite

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“…Over 66 % of fish sub lots have been accepted as they present less than one parasite in the flesh, expressed as MA [24] (Table 1). MA, due to its correlation with P and with the number of samples [25], could be used to estimate the degree of infestation [26], particurarly in the case of fishery products sold in batches. With regard to the naked eye rejected products, 33.33% of the total rejections belong to FAO 27.…”

Section: Parasite Identificationmentioning

confidence: 99%

Risk-based scoring and genetic identification for anisakids in frozen fish products from Atlantic FAO areas

et al. 2020

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Background: The presence of Anisakis larvae in fish represents a major public health concern. Effective risk management procedures should be applied to prevent heavily infected products from reaching the market. The aim of the study is to provide preliminary data on parasite exposure and risk classification in frozen fish products by applying a risk categorization scheme (site, abundance, density and epidemiology -SADE) and Fish Parasite Rating (FPR) method. Fish and cephalopods samples (N = 771) from 5 different FAO Atlantic areas were examined and categorized after an accurate visual inspection and a chloro-peptic digestion.Results: In 25 out of 33 fish species parasite larvae were found. 10897 anisakids larvae were collected and identified to genus level. Molva dypterygia, Conger conger, Zeus faber and Aphanopus carbo were shown to be the most highly infected species. SADE and FPR scores were 1 and poor, respectively, for the referred species, because of the disseminated Anisakis infection and commercial rejection. Conclusion: SADE/FPR method showed high specificity and accuracy. The information provided in this work could be used in early warning systems for the detection of parasites in fishery products and might help fishing industries in establishing management strategies for infected stocks in terms of cost saving decisions.

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Estimation of the number of Anisakis larvae in commercial fish using a descriptive model based on real‐time PCR

et al. 2020

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BACKGROUNDSeafood parasitation by Anisakis (Anisakidae) larvae has been reported in most of the oceans and seas worldwide. The presence of these nematodes in commonly consumed fish represents a potential hazard for consumers as they can provoke gastrointestinal symptoms and allergic reactions. In the present work, the capacity of a SYBR Green qPCR protocol to quantify Anisakis larvae in commercial fish was evaluated using experimentally spiked samples with different numbers (0–50) of A. simplex third‐stage larvae (L3). To verify the agreement of the obtained results, 25 naturally infected fish specimens of Atlantic blue whiting underwent a parallel visual inspection.RESULTSThe logarithmic behavior of the Cq data obtained from the experimentally spiked samples allowed the development of a descriptive mathematical model that correlates the Cq value with the number of Anisakis larvae (R2 = 0.9908, CV = 2.37%). In the commercial blue whiting specimens there was a high correlation between the results of the molecular technique and the visual inspection (R2 = 0.9912); the Bland–Altman analysis showed that 94% of the differences were within the limits of agreement (−4.98 and 6.68), indicating the reliability of the descriptive mathematical model based on the SYBR Green qPCR technique.CONCLUSIONThe descriptive function presented based on the SYBR Green qPCR assay is promising as a sensitive and accurate tool for measuring the Anisakis larval load in commercial fish, with a potential application not only in the food industry but also in prevention programs for public health. © 2020 Society of Chemical Industry

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Assessment of a Sampling Plan Based on Visual Inspection for the Detection of Anisakid Larvae in Fresh Anchovies (Engraulis encrasicolus). A First Step Towards Official Validation?

Cited by 18 publications

References 39 publications

Anisakis spp. larvae in different kinds of ready to eat products made of anchovies ( Engraulis encrasicolus ) sold in Italian supermarkets

Anisakis spp. larvae in different kinds of ready to eat products made of anchovies ( Engraulis encrasicolus ) sold in Italian supermarkets

Risk-based scoring and genetic identification for anisakids in frozen fish products from Atlantic FAO areas

Estimation of the number of Anisakis larvae in commercial fish using a descriptive model based on real‐time PCR

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