Spelt products are popular with consumers achieving higher market prizes, making them susceptible to food adulteration with less valuable cereals. To facilitate product control, a recently developed duplex droplet digital PCR method enables the detection and quantification of common wheat (Triticum aestivum) contaminations in food products made from spelt (Triticum spelta). The duplex droplet digital PCR assay targets the γ-gliadin gene and the Q-locus of both subspecies. In this study, the method was validated in an interlaboratory ring trial involving 11 participating laboratories. Test materials containing defined proportions of spelt and common wheat were prepared and tested. The ring trial procedure included DNA extraction of the test samples and determination of subspecies proportions using droplet digital PCR. Results from the ring trial confirmed the method's capability for specific detection and quantification of common wheat in spelt, with acceptable relative measurement uncertainties, and without requiring reference material for calibration. To our knowledge, this is the first interlaboratory validation of a digital PCR method for species differentiation in food.

Tuna cans are relevant seafood products for which mixtures of different tuna species are not allowed according to European regulations. In order to support the prevention of food fraud and mislabelling, a next-generation sequencing methodology based on mitochondrial cytochrome b and control region markers has been tested. Analyses of defined mixtures of DNA, fresh tissue and canned tissue revealed a qualitative and, to some extent, semiquantitative identification of tuna species. While the choice of the bioinformatic pipeline had no influence in the results (p = 0.71), quantitative differences occurred depending on the treatment of the sample, marker, species, and mixture (p < 0.01). The results revealed that matrix-specific calibrators or normalization models should also be used in NGS. The method represents an important step towards a semiquantitative method for routine control of this analytically challenging food matrix. Tests of commercial samples uncovered mixed species in some cans, being not in compliance with EU regulations.

The increasing consumption of seafood products raises concerns over their sustainability and the conservation of marine resources. Seafood traceability, enabled by a regulated labelling system, is important to prevent overexploitation of these resources. The regulations (EU) No.1169/2011 and (EU) No 1379/2013 are the European legislative tools that specify the mandatory information that must be present on seafood labels. The present study analysed the labels of seafood products sold in different European countries in order to verify the presence of mandatory information required by EU regulations currently in place. The results show that there is a difference in compliance among groups of products and among countries. The country with the lowest level of compliance was The United Kingdom (still part of EU when the study was carried out), with an overall compliance of 63.7%. The country with the highest level of compliance was Portugal (87.2%). Across all the countries analysed, supermarkets were more compliant than fishmonger's shops and Processed Prepacked products were more conformed best to the EU labelling legislation when compared to Unprocessed Non-Prepacked products. Differences among different areas of the same country were also observed. Fishing gear, scientific name, fishing/production method and date of freezing were the types of information most frequently missing on the labels examined. The results of this study pose the bases for further actions, that can be taken by relevant institutions, to improve compliance throughout the supply chain.

Seafood products are particularly vulnerable to food fraud and mislabelling, which may have negative implications on fisheries resources, economy, consumer health and trust. This is a considered problem for scallops, since these high-valued seafood products are usually sold without their morphologically characteristic shells. Scallop products differ in taste and value and usually species of the genus Pecten spp. are especially expensive in many European countries. The aim of the present study was the development of a multiplex TaqMan real-time PCR assay that allows a rapid and reliable authentication of the three commercially important species/genera Pecten spp. (usually King scallop P. maximus), Atlantic sea scallop Placopecten magellanicus, and Japanese scallop Mizuhopecten yessoensis. The design of primers and probes was based on mitochondrial 16S rRNA gene amplifying fragments of 138-198 bp. Following the optimization of the multiplex real-time PCR assay, analyses on efficiency, limit of detection, specificity, robustness, and crosstalk were conducted for validation purpose. Average Cq values of 20 ng DNA obtained for Pecten spp. were 17.64 +/- 0.89, for P. magellanicus 18.42 +/- 0.83, and for M. yessoensis 17.08 +/- 0.79. Non-target species produced either no fluorescence signal or the Cq differed significantly from those of target species (p < 0.01). Finally, the newly developed real-time PCR assay was tested on commercial samples from German supermarkets and fishmongers accompanied by simultaneous verification through Sanger sequencing, which revealed a high mislabelling rate of 48%, especially for products purchased at fishmongers. The results emphasize the need of a control method that allows the rapid analysis of sufficient sample quantities. The study is one of the first which presents a multiplex TaqMan real-time PCR approach for the authentication of commercially important seafood species.

Anisakid nematode larvae (NL) in fish products comprise a risk to human health and, if visible, lead to the rejection of these products by consumers. Therefore, great efforts are being made for the identification of these anisakid larvae to estimate the potential consumer health risk as well as to develop effective detection methods in order to prevent the introduction of heavily infected fish products into the market. The tasks of national reference laboratories include the improvement of detection methods and to promote their further development. As a prerequisite for improved detection, it is important to understand the structural properties of anisakid NL and compounds produced during host-parasite interactions. This review provides an overview of the intrinsic properties of anisakid NL and reports the latest detection methods in published literature. First, in order to define the potentially interesting intrinsic properties of anisakid nematodes for their detection, anatomy and compounds involved in host-parasite interactions are summarised. These can be used for various detection approaches, such as in the medical field or for allergen detection in fish products. In addition, fluorescence characteristics and their use as both established and promising candidates for detection methods, especially in the field of optical sensing technologies, are presented. Finally, different detection and identification methods applied by the fish processing industries and by control laboratories are listed. The review intends to highlight trends and provide suggestions for the development of improved detection and identification methods of anisakid NL in fish products.

Due to its remote and isolated location, Antarctica is home to a unique diversity of species. The harsh conditions have shaped a primarily highly adapted endemic fauna. This includes the notothenioid family Channichthyidae. Their exceptional physiological adaptations have made this family of icefish the focus of many studies. However, studies on their ecology, especially on their parasite fauna, are comparatively rare. Parasites, directly linked to the food chain, can function as biological indicators and provide valuable information on host ecology (e.g., trophic interactions) even in remote habitats with limited accessibility, such as the Southern Ocean. In the present study, channichthyid fish (Champsocephalus gunnari: n = 25, Chaenodraco wilsoni: n = 33, Neopagetopsis ionah: n = 3, Pagetopsis macropterus: n = 4, Pseudochaenichthys georgianus: n = 15) were collected off South Shetland Island, Elephant Island, and the tip of the Antarctic Peninsula (CCAML statistical subarea 48.1). The parasite fauna consisted of 14 genera and 15 species, belonging to the six taxonomic groups including Digenea (four species), Nematoda (four), Cestoda (two), Acanthocephala (one), Hirudinea (three), and Copepoda (one). The stomach contents were less diverse with only Crustacea (Euphausiacea, Amphipoda) recovered from all examined fishes. Overall, 15 new parasite-host records could be established, and possibly a undescribed genotype or even species might exist among the nematodes.

Canned liver is a highly valuable and healthy product from the important commercial fish species cod Gadus morhua. Due to high levels of contaminants such as dioxins and dioxin-like PCBs (polychlorinated biphenyls) in cod liver products from the Baltic Sea, the grounds for supply of cod liver were relocated to areas around Iceland in the Irminger Sea with lower levels of contaminants. In the present study, 62 canned samples, as well as 132 fresh cod livers from Greenland and Iceland, were analysed in order to assess the infection levels with anisakid nematode species. High prevalence and abundance of the potentially zoonotic nematode species Anisakis simplex s.s. and Pseudoterranova decipiens s.s. were found in the fresh livers. Lower infection levels in canned livers compared to fresh livers might indicate successful industrial processing measures in order to reduce nematode larvae in the final products. However, two heat stable allergens, Ani s 1 and Ani s 4, were verified from isolated nematode larvae from canned products. The effectiveness of methods to further reduce the number of nematode larvae during industrial processing of cans with cod liver is discussed.

Abstract. Although coccolithophore physiological responses to CO2-induced changes in seawater carbonate chemistry have been widely studied in the past, there is limited knowledge on the variability of physiological responses between populations from different areas. In the present study, we investigated the specific responses of growth, particulate organic (POC) and inorganic carbon (PIC) production rates of three populations of the coccolithophore Emiliania huxleyi from three regions in the North Atlantic Ocean (Azores: six strains, Canary Islands: five strains, and Norwegian coast near Bergen: six strains) to a CO2 partial pressure (pCO2) range from 120 to 2630 µatm. Physiological rates of each population and individual strain increased with rising pCO2 levels, reached a maximum and declined thereafter. Optimal pCO2 for growth, POC production rates, and tolerance to low pH (i.e., high proton concentration) was significantly higher in an E. huxleyi population isolated from the Norwegian coast than in those isolated near the Azores and Canary Islands. This may be due to the large environmental variability including large pCO2 and pH fluctuations in coastal waters off Bergen compared to the rather stable oceanic conditions at the other two sites. Maximum growth and POC production rates of the Azores and Bergen populations were similar and significantly higher than that of the Canary Islands population. This pattern could be driven by temperature–CO2 interactions where the chosen incubation temperature (16 ∘C) was slightly below what strains isolated near the Canary Islands normally experience. Our results indicate adaptation of E. huxleyi to their local environmental conditions and the existence of distinct E. huxleyi populations. Within each population, different growth, POC, and PIC production rates at different pCO2 levels indicated strain-specific phenotypic plasticity. Accounting for this variability is important to understand how or whether E. huxleyi might adapt to rising CO2 levels.

Eastern boundary upwelling provides the conditions for high marine productivity in the Canary Current System off NW-Africa. Despite its considerable importance to fisheries, knowledge on this marine ecosystem is only limited. Here, parasites were used as indicators to gain insight into the host ecology and food web of two pelagic fish species, the commercially important species Trichiurus lepturus Linnaeus, 1758, and Nealotus tripes Johnson, 1865. Fish specimens of T. lepturus (n = 104) and N. tripes (n = 91), sampled from the Canary Current System off the Senegalese coast and Cape Verde Islands, were examined, collecting data on their biometrics, diet and parasitisation. In this study, the first parasitological data on N. tripes are presented. T. lepturus mainly preyed on small pelagic Crustacea and the diet of N. tripes was dominated by small mesopelagic Teleostei. Both host species were infested by mostly generalist parasites. The parasite fauna of T. lepturus consisted of at least nine different species belonging to six taxonomic groups, with a less diverse fauna of ectoparasites and cestodes in comparison to studies in other coastal ecosystems (Brazil Current and Kuriosho Current). The zoonotic nematode Anisakis pegreffii occurred in 23% of the samples and could pose a risk regarding food safety. The parasite fauna of N. tripes was composed of at least thirteen species from seven different taxonomic groups. Its most common parasites were digenean ovigerous metacercariae, larval cestodes and a monogenean species (Diclidophoridae). The observed patterns of parasitisation in both host species indicate their trophic relationships and are typical for mesopredators from the subtropical epi- and mesopelagic. The parasite fauna, containing few dominant species with a high abundance, represents the typical species composition of an eastern boundary upwelling ecosystem.

The Atlantic herring is a schooling, pelagic species that inhabits both sides of the North Atlantic Ocean. Herring stock identification is usually based on several approaches, including fish meristic characters, population genetic analysis and the use of parasite species composition. A total of 654 Anisakis spp. larvae collected from herring of four fishing grounds in the Norwegian Sea, Baltic Sea, North Sea, and the English Channel off the French coast, was identified to species level using diagnostic allozymes and sequence analysis of EF1 alpha - 1 nDNA and the mtDNA cox2 genes. Population genetic differentiation of Anisakis simplex (s. s.) among the different fishing areas was estimated, at the intraspecific level, on the basis of mtDNA cox2 sequences analysis. Spatial comparison based on molecular variance analysis and F values was performed for the collected specimens (among regions). Haplotype network construction showed relevant differences in haplotype frequencies between samples of A. simplex (s. s.) from the different geographical areas. Results indicate a genetic sub-structuring of A. simplex (s. s.) obtained from herring in different areas, with the population from the Norwegian Sea being the most differentiated one, and with North Sea and Baltic Sea populations being most similar. The population genetic structure of A. simplex (s. s.) was in accordance with the herring population genetic structure throughout the host's geographical range in the NE Atlantic. Results suggest that mtDNA cox2 is a suitable genetic marker for A. simplex (s. s.) population genetic structure analysis and a valuable tool to elucidate the herring stock structure in the NE Atlantic Ocean.

Monogeneans are typically ectoparasites characterized by a direct life cycle, cosmopolitan distribution, great biodiversity, and high host specificity. Special adaptations enable their existence under extreme abiotic conditions in Antarctic waters. Existing literature was compiled to provide an overview on Antarctic monogenean biodiversity, host specificity, and biogeography. Twenty three monogenean parasite species of 7 families and 25 fish host species of 3 families were documented. Overall, host specificity was high with the subclass Monopisthocotylea infecting primarily fish of the suborder Notothenoidei while Polyopisthocotylea parasitized exclusively on Macrouridae. The high degree of host specificity was associated with special adaptations of the opisthaptor and reproduction. Gyrodactylidae were considered to be especially successful in cold-water habitats as a result of their viviparous reproductive mode. Due to the scarcity of studies on Antarctic monogeneans, hypotheses on host specificity and biogeography may rather be suggestive and their biodiversity be underestimated.

The main commercial fish species in the Irminger Sea, the beaked redfish Sebastes mentella, is commonly infected with the copepod Sphyrion lumpi. This ectoparasite is often used as a biological marker for stock discrimination to evaluate the still-debated metapopulation structure of beaked redfish. Nevertheless, it is still not understood whether parasite abundances and communities vary over longer time periods. In the present study, we investigated the abundance of S. lumpi in S. mentella of the pelagic zone of the Irminger Sea and adjacent waters. Our analyses revealed that live S. lumpi abundance remained constant during summer over a sampling period from 2001 to 2015, which confirms its validity as a biomarker. As S. mentella forms dense aggregations during mating, our results suggest that host densities are, regardless of a fishery-induced decrease in host biomass, large enough to facilitate the direct transmission of S. lumpi. After correcting for covariate effects, two stock units could be differentiated, which supports a continuation of the current fishery management strategy.