Development and Application of Novel Chemiluminescence Immunoassays for Highly Sensitive Detection of Anisakis simplex Proteins in Thermally Processed Seafood.

The third-stage larvae (L3) of Anisakis simplex are the most important source of hidden allergens in seafood products. However, there exist no commercial methods for detecting Anisakis proteins in food. Furthermore, only a few methods have been validated for the detection of A. simplex in thermally processed food. The aims of our study are (i) the development and validation of high-sensitivity chemiluminescent (CL) immunoassays for the detection of A. simplex proteins in processed seafood, (ii) and A. simplex antigen detection in common seafood products from Polish markets. We developed and validated CL sandwich ELISA (S-ELISA) and CL competitive ELISA (C-ELISA) methods for A. simplex proteins detection in food, with respective detection limits of 0.5 and 5 ng/mL. The usefulness of the assays for detecting A. simplex proteins in highly processed food was evaluated by examination of autoclaved canned fish spiked with A. simplex larvae (1-8 larvae/200 g). Commercial real-time PCR was unable to detect A. simplex in autoclaved samples at all levels of enrichment with Anisakis larvae. CL-S-ELISA was used to test various types of seafood products from Polish markets. Among all tested products (n = 259), 28% were positive. A. simplex antigens were found mostly (n = 39) in smoked fish products: mackerel, herring, cod, and hake. Other positive samples were found in marinated herrings, canned cod livers, canned mackerels, and surimi sticks. In tuna, Atlantic argentine, anchovy, sardine, sprat, and squid products, A. simplex antigens were not detected. This study provides novel effective tools for the detection of A. simplex proteins in processed food and highlights the potential allergic hazards for Anisakis-sensitized Polish consumers of seafood.

Occurrence of Trichinella spp. in rats on pig farms.

INTRODUCTION: The highest risk of trichinellosis for human is considered in eating meat products containing live larvae, mostly from wild boars or pigs. Spreading of Trichinella spp. may occur in various ways, one of which is transmission by vectors. The rat is considered to be the most common vector for Trichinella parasite. The population of rats living on pig farms can play an important role in maintaining or spreading the parasite to other animals. OBJECTIVE: The aim of presented survey was to investigate the occurrence of Trichinella spp. in rats on farms with pigs infected with this parasite. MATERIAL AND METHODS: From pig farms selected for study, the muscles of collected rats were investigated by magnetic stirrer digestion method to assess occurrence of Trichinella in the rat population. Isolated Trichinella parasites were identified under stereomicroscope and multiplex PCR were performed for species identification. RESULTS: Rats infected with Trichinella spp. were discovered on three of five investigated pig farms. The mean extent of invasion in rats from the studied farms was 23.33%. The calculated medium intensity of invasion was 4.09 lpg (larvae per gram) (SD 5.41). All larvae of Trichinella discovered from rats were identified as T.spiralis. CONCLUSIONS: The results obtained indicate that in farms with a high prevalence of Trichinella invasion in pigs there are very likely to be found rats infected by this nematode. This suggests possibility to maintain the invasion in herd and spread into neighborhood farms.

First case of Trichinella nativa infection in wild boar in Central Europe-molecular characterization of the parasite.

The examination of wild boars gained in Poland shows for the first time occurrence of Trichinella nativa, freeze-resistant species of Trichinella in this host from the central Europe region. This finding is not only one of several cases of T. nativa invasion in wild boars all over the world but also one of the very few cases of T. nativa detected so far beyond the known boundary of occurrence of this species. The molecular characterization of discovered larvae based on analysis of partial genes: 5s rDNA-ISR and CO1 confirm the findings. Moreover, the analyzed DNA sequences of both genes present new haplotypes of T. nativa in comparison to that described previously.