High occurrence of Anisakidae at retail level in cod (Gadus morhua) belly flaps and the impact of extensive candling

The presence of Anisakidae at retail level, after the routine screening via candling, was investigated in cod, the most commonly consumed fish species in Belgium. A total of 780 pre-packed belly flap samples destined for one branch of retail shops were collected from a Belgian wholesale company. To recover all larvae, each sample was first candled and thereafter enzymatically digested. Larvae were morphologically identified to the genus level and a subset was additionally molecularly confirmed by amplification of the ITS fragment and HinfI/HhaI enzyme restriction. The PCR/RFLP profiles of Contracaecum spp. were determined and confirmed with sequencing by the European Reference Laboratory for Parasites (Istituto Superiore di Sanità). The positivity rate of Anisakidae in the individual cod samples was 18% [95%-CI: 15–21%], with a mean intensity of one larva [range: 1–6]. Belly flaps were sold packed primarily by two, with a one-in-three chance of buying an infected package. Pseudoterranova spp. infections (single infections) were most frequently detected (positivity rate 9% [95%-CI: 7–11]), closely followed by Anisakis spp. (7% [95%-CI: 6–9]). Co-infections of Pseudoterranova spp. and Anisakis spp. comprised 8% of the infections, with a positivity rate of 1% [95%-CI: 1–3%]. All belly flaps reportedly were candled prior to our sampling, nonetheless our results indicated that an additional candling screening before packaging would identify an extra third of the infections and larvae. In 19 of the 139 infected samples, all larvae were recovered by the additional candling, thereby removing the infection risk for consumers. In conclusion, this study shows that cod belly flaps infected with zoonotic parasites reach the Belgian consumer. Although a second candling step at retail level could be helpful in reducing the consumer risk, additional measures are needed since 66% of infections would still remain undetected

Ascaridoid nematodes in fish : occurrence, diversity, and evaluation of candling towards exploring the public health relevance in Belgium

Ascaridoids are marine nematodes occurring in a wide range of fish hosts. In the intermediate fish host, L3 (third stage) larvae reside in the viscera (primary infection site) and can migrate, both post- and ante-mortem to the fish musculature. Humans are accidental dead-end host, and may develop gastro-intestinal complaints after ingestion of a viable larva, or acquire allergic symptoms even after ingestion of a dead larva or residual allergens. The industry is tasked with the detection and removal of these larvae, in absence of a fast, cheap, and highly sensitive detection technique. Remaining larvae may, besides representing a food safety problem, trigger consumers complaints. The objectives of this thesis were to estimate the occurrence of ascaridoids in fish destined for the Belgian market; and to assess the sensitivity of the current mostly used diagnostic tool in fish, namely candling

Presence of anisakidae in commercial fish species imported into the Belgian food markets : a systematic review and meta-analyses

Anisakidae are marine zoonotic nematodes with most commercial fish species as intermediate hosts. Both public health risks and socio-economic problems are attributed to these larvae. Despite these concerns, the occurrence of Anisakidae in commercial fish species in Belgium remains unknown. Therefore, the main objective of this systematic review was to look into studies assessing the prevalence and intensity (level of infection) of Anisakidae in countries importing fish to the Belgian market. The databases of PubMed, Web of Science, Cordis, Google Scholar, Google, African Journals online and Asia Journals online were searched. Main eligibility criteria were: fish species consumed in Belgium; studies conducted in one of the main importing countries; and the availability of prevalence data. From the original 519 identified studies, 83 were included with data from Spain, Germany, Chile, Denmark, Turkey, France, China, England, Belgium, Norway, Iceland, Senegal and Sweden. Overall results show a widespread occurrence of Anisakidae with a high variability in prevalence between fish species and fishing sea. Cod (Gadus morhua) and Atlantic salmon (Salmo solar), the most consumed fish species in Belgium, have a mean prevalence of 33% and 5% respectively. Of all investigated fishing zones, fish caught in the Northeast Atlantic has the highest rate of infection (68%). Furthermore, higher prevalences were found when looking at the viscera (mean prevalence 59%) compared to the muscle (29%) and with superior techniques such as enzymatic digestion or UV press (46%) compared to candling, the routine method (23%). Farmed fish were found to be the least infected (2%) but were still not Anisakidae free. The widespread presence of Anisakidae and the associated food safety implications indicate the need to further investigate the presence of Anisakidae in fish in the Belgian market

Ascaridoids in commercial fish : occurrence, intensity and localization in whole fish and fillets destined for the Belgian market

Anisakidae and Raphidascaridae are marine nematodes present in a wide range of fish hosts, which may cause gastro-intestinal complaints and/or allergy in human, in addition to economic losses for the industry. Data regarding the presence of these parasites in fish for the Belgian market is currently missing; therefore, our aim was to investigate the presence and intensity of ascaridoids in a wide range of commercially fish species. A total of 415 fish samples, belonging to 36 different fish species, were collected from a Belgian whole-sale company. Ascaridoid larvae from the viscera (if present) and the muscles were collected by enzymatic digestion and the prevalence, median intensity, mean number of larvae per 100 g infected muscle, and localisation were determined. An overall prevalence of 53% [95%-CI: 42–63%] in the viscera and 27% [95%-CI: 23–32%] in the muscles was observed. Infection in the muscles varied between the fish species; no larvae were detected in 13 fish species, while a high prevalence (>78%) was observed in pollack, halibut, and gurnard. Most samples originated from the Northeast Atlantic Ocean, with the highest prevalence in the muscles observed in the Barents & Norwegian Sea (65% [95%-CI: 38–86]). Muscle samples were, if possible, divided in an anterior region, belly flap, medial region, and posterior region, with the most infections and larvae found in the belly flaps. In all samples, a total of 2569 larvae were recovered, with 1594 larvae originating from the viscera and 975 from the muscles; with an average of two larvae per 100 g infected fillet detected. Larvae were morphologically identified, and a subgroup was further confirmed using PCR/RFLP, resulting ultimately in the identification of Anisakis simplex s.s. (1853 larvae), A. pegreffii (137), A. simplex/pegreffii hybrid genotype (38), Pseudoterranova decipiens (160) and Hysterothylacium aduncum (380). This study demonstrates that ascaridoid larvae are highly prevalent in different fish species on the Belgian market

Sensitivity of candling as routine method for the detection and recovery of ascaridoids in commercial fish fillets

Ascaridoids are one of the main parasitic hazards in commercial fish. Candling is the current industrial screening method whereby visible ascaridoid larvae are detected on a light table and manually removed. The aim of this study was to assess the sensitivity (Se) and negative predictive value (NPV) of this method. To make targeted recommendations to the fish industry, the Se was calculated per fish part, larval genus, and fish species. All fish parts (n = 615) were first candled, and larvae were collected, followed by enzymatic digestion to recover the remaining larvae. A fish part was considered positive if at least one larva was detected using candling and/or enzymatic digestion, with both methods combined as reference standard. The overall Se of candling was 31% (95% CI 23-41%) and NPV was 87% (95% CI 85-90%). The Se increased with higher numbers of larvae/100 g infected muscle. A low NPV was found for the belly flaps, therefore we either advise the removal or proper freezing of this part. Lastly, the Se and larval recovery was the highest for the darker and larger Pseudoterranova spp. larvae. Due to the low overall efficacy of candling, further assessment of its cost-benefit and impact on consumers' health risk should be conducted