Does Anisakis spp. infestation affect the proximate composition, fatty acids, and minerals contents of its host Merluccius merlucccius?

Fish play an important role in human nutrition. They are not only a great source of protein and healthy fats, but also a unique source of essential nutrients such as omega-3 fatty acids. Moreover, most fish are parasitized, and some of these parasites are able to influence the reallocation of resources in their favor and thus reduce the nutritional quality of the fish. The present study was conducted to investigate the impact of the third stage larvae (L3) of Anisakis spp. on the proximate composition, macro-minerals (potassium, calcium, and sodium), and fatty acids of European hake (Merluccius merluccius Linnaeus, 1758). In parasitized female group, our results revealed a decrease (p< 0.005) in the amount of carbohydrate by 6.5%, of calcium by 17%, and of 2 polyunsaturated fatty acids (arachidonic acid (C20: 4w-6), and eicosapentaenoic acid (C20: 5w-3) with 33% and 15% respectively. Simultaneously, an increase by 25% in the level of a single saturated fatty acid C10:0 was noticed. According to the principal component analysis, the parasitized female was wealthy of saturated fatty acids and monounsaturated fatty acids and contains less of polyunsaturated fatty acids, omega-3 fatty acids, and omega-6 fatty acids than the unparasitized female and male. No significant changes were observed in the biochemical composition of male hake, probably due to the low mean intensity of L3 larvae of Anisakis spp. in this group.

Diclidophora luscae (Monogenea: Diclidophoridae) in pouting, Trisopterus luscus (Linnaeus, 1758) from the northeast Atlantic; epidemiology, morphology, molecular and phylogenetic analysis.

Diclidophora (Monogenea) species are gill parasites with a stenoxenic specificity occurring only in Gadiformes. Epidemiological, morphological, molecular and phylogenetic studies were performed on 594 Diclidophora specimens collected from 213 Trisopterus luscus captured in the northeast Atlantic off the Portuguese coast during 2012, 2013 and 2020. Prevalence, parasite abundance and infection intensity were determined. Positive correlation between fish weight and length and infection intensity was observed. The effects of preservation on the parasite morphological features were studied, highlighting that specimen's identification should be reinforced by molecular studies. A sequence of D. luscae capelanii from T. capelanus captured in the Mediterranean Sea included in the 28S rDNA molecular analysis was nested within a robust D. luscae clade. Data analysis suggested that this species is in fact D. luscae, which is compatible with T. luscus and T. capelanus. The identity of fish hosts was confirmed by barcoding. For the first time, data on the infection parameters is shown, highlighting the importance of including this parasite in the monitoring plans for a holistic approach with possible effects for the management of pouting resources aiming of attaining sustainable development and biodiversity conservation measures, according to the 14th objective of the 2030 agenda.

Optimization of tools for the detection and identification of Cryptocotyle metacercariae in fish: Digestion method and viability studies.

Some trematode metacercariae, including marine digeneans belonging to the genus Cryptocotyle, induce black spots in target tissues due to the attraction of fish host melanophores. To promote precise quantification of infection, the counting of black spots has to be confirmed by reliable quantification of metacercariae after tissue digestion. This process ensures the isolation of undamaged parasites for morphological and molecular identification. The aim of this work was to optimize the pepsin digestion protocol and to assess the duration of viability of Cryptocotyle metacercariae in fish post-mortem (pm). Four digestion protocols were compared by measuring the viability rate of metacercariae. The present study shows that the orbital digestion method was the least destructive for metacercariae and allowed better quantification of Cryptocotyle infection. Moreover, morphological identification seemed reliable up to 8 days pm for Cryptocotyle infection.

Polar and neutral lipid composition of the copepod Lernaeocera lusci and its host Merluccius merluccius in relationship with the parasite intensity.

Parasitic copepod Lernaeocera lusci is a common mesoparasite of the hake Merluccius merluccius. Although widely distributed throughout the Mediterranean, little is known about this pathogen. The current study was designed to assess the impact of different L. lusci infection loads on lipid classes and their fatty acid (FA) composition in both parasite and the host organs (gills, liver, and muscle). Results showed a significant decrease in total lipid, neutral lipid (NL), and polar lipid (PL) contents in all analyzed host's organs in relationship with parasite intensity. Gills appeared to be the most impacted organ under the lowest parasite intensity (loss of 50% of NL and PL amounts). At the highest parasitic infection, a loss of about 80% of lipid moieties was recorded in all analyzed organs. Simultaneously, no significant differences were found for the parasite reflecting its ability to sustain an appropriate lipid amount required for its survival and development. Significant changes in the FA composition were recorded in both host and parasite. Particularly, we have noticed that for L. lusci, the intraspecific competition has resulted in an increased level of some essential FA such as C22:6n-3 (docosahexaenoic acid, DHA), C20:5n-3 (eicosapentaenoic acid, EPA), and C20:4n-6 (arachidonic acid, ARA). This probably reflects that in addition to a direct host FA diversion, L. Lusci can modulate its FA composition by increasing the activity of desaturation. Within the host, liver PL appeared to be the less impacted fraction which may mirror an adaptive strategy adopted by the host to preserve the structural and functional integrity of this vital organ.

Anisakis simplex (s.l.) resistance to the action of gastric enzymes depends upon previous treatments applied to infected fish mince and affects antigen release.

BACKGROUND: Freezing is considered the most suitable technological treatment to avoid Anisakis infection from eating raw or undercooked fish but modifications of their cuticles upon freezing may reduce their resistance to gastric fluids, provoking a greater release of allergens. This work aimed to study the relationship between freezing-induced modifications of Anisakis simplex s.l., antigen recognition, and resistance to oral and gastric digestion in spiked fish mince. RESULTS: (i) Differences between non-treated larvae and larvae that survived freezing / thawing were studied in terms of respiratory capacity, survival in simulated gastric fluid (SGF), recognition of antigens and allergens. (ii) Untreated (i.e. chilled) mince containing live larvae, mince frozen at two freezing rates, with a negative (uninfected) mince and a positive mince (infected with broken larvae) as controls, were subjected to the oral and gastric phases of a simulated digestion process. Anisakis able to survive freezing showed lower resistance to gastric fluid (i.e. faster mortality as compared to controls). Untreated larvae released significantly more antigens than freeze-surviving larvae but only after 96 h in SGF. In treatments rendering complete larvae mortality, the highest loss of larvae integrity was found upon fast freezing. There was a positive correlation between antigen release and the number of ruptures of larvae after the oral digestion phase, whereas a more complex trend was observed after oral plus gastric digestion phases. CONCLUSION: These results suggest a new factor to consider for sensitized patients and suggest that the numbers of L3 should be reduced before industrial freezing to minimize risk. © 2020 Society of Chemical Industry.

Immunohistopathological response against anisakid nematode larvae and a coccidian in Micromesistius poutassou from NE Atlantic waters.

A survey on Anisakis simplex (sensu stricto (s.s.)) from blue whiting, Micromesistius poutassou, in the north-eastern Atlantic Ocean revealed the occurrence of high infection levels of third larval stages in visceral organs and flesh. Larvae were genetically identified with a multilocus approach as A. simplex (s.s.). Histochemical, immunohistochemical and ultrastructural observations were conducted on 30 M. poutassou specimens. Gonads, pyloric caeca and flesh harboured encapsulated larvae of A. simplex (s.s.) but no intense host reaction was encountered around the parasite in the above organs. In the liver, the most infected organ, the larvae co-occurred with the coccidian Goussia sp. Within the granuloma around the A. simplex (s.s.) larvae, two concentric layers were recognized, an inner mostly comprising electron-dense epithelioid cells and an outer layer made of less electron-dense epithelioid cells. Macrophages and macrophage aggregates (MAs) were abundant out of the granulomas, scattered in parenchyma, and inside the MAs, the presence of engulfed Goussia sp. was frequent. In liver tissue co-infected with Goussia sp. and A. simplex (s.s.), hepatocytes showed cytoplasmic rarefaction and acute cell swelling. Results suggest that the host-induced encapsulation of A. simplex (s.s.) larvae is a strategic compromise to minimize collateral tissue damage around the larval infection sites, to facilitate the survival of both parasite and host.

Effects of Goussia infecting the blue whiting and phylogenetic placement of Goussia infecting marine fish off Northern Portugal.

Coccidian parasites of fish have received considerably less attention than their terrestrial counterparts, and within piscine hosts, most studies have focused on freshwater fish. The present study aimed to describe oocyst morphology, phylogenetic affinities, and the impacts of coccidian parasites infecting the internal organs of a commercially valuable marine fish, the blue whiting (Micromesistius poutassou), captured off the Portuguese coast. As part of the phylogenetic analysis, sequences from coccidians infecting the pout (Trisopterus luscus) and the Atlantic chub mackerel (Scomber colias) were included, and the oocyst morphology of the coccidians infecting the former was also reported. Results showed that the prevalence of coccidiosis in the blue whiting was very high (> 82%), occurring in all analyzed organs, despite being more abundant in the liver. A significant negative correlation was found between the abundance of the parasites in the liver and host condition index (p < 0.05), which indicates a negative effect on the fitness of this host. Phylogenetic analyses of the parasites found in all three species examined identified three different species of Goussia, closely related to Goussia clupearum. Adding to previous research, we propose the existence of a fourth group of Goussia, the clupearum type, able to infect multiple organs and phylogenetic related with G. clupearum.

Phylogenetic position of deep-sea opecoelid digenean Tellervotrema beringi (Mamaev, 1965) (Trematoda: Opecoelidae) based on novel genetic data.

Adult opecoelid digeneans consistent with Tellervotrema beringi (Mamaev, 1965) were found in the intestine of the deep-water fish Antimora microlepis Bean collected off the northern Kuril Islands (Russia). Earlier, T. beringi was recorded only in macrourid fish. This is the first record of a species of Tellervotrema Gibson & Bray, 1982 from fishes of the family Moridae (Gadiformes). A partial 28S rRNA gene sequence was obtained for this species and was compared with the available sequences on GenBank from other opecoelids. The phylogenetic analysis did not support the current taxonomic hypothesis that the genus Tellervotrema belongs to the subfamily Podocotylinae Dollfus, 1959. Our results demonstrated that species of this genus formed a clade with those of the genera Mesobathylebouria Martin, Huston, Cutmore & Cribb, 2019 and Abyssopedunculus Martin, Huston, Cutmore & Cribb, 2019. The morphological synapomorphies of the Tellervotrema + (Mesobathylebouria + Abyssopedunculus) clade are currently unclear, and this group does not have an adequate subfamilial classification.

Functional insights into the infective larval stage of Anisakis simplex s.s., Anisakis pegreffii and their hybrids based on gene expression patterns.

BACKGROUND: Anisakis simplex sensu stricto and Anisakis pegreffii are sibling species of nematodes parasitic on marine mammals. Zoonotic human infection with third stage infective larvae causes anisakiasis, a debilitating and potentially fatal disease. These 2 species show evidence of hybridisation in geographical areas where they are sympatric. How the species and their hybrids differ is still poorly understood. RESULTS: Third stage larvae of Anisakis simplex s.s., Anisakis pegreffii and hybrids were sampled from Merluccius merluccius (Teleosti) hosts captured in waters of the FAO 27 geographical area. Specimens of each species and hybrids were distinguished with a diagnostic genetic marker (ITS). RNA was extracted from pools of 10 individuals of each taxon. Transcriptomes were generated using Illumina RNA-Seq, and assembled de novo. A joint assembly (here called merged transcriptome) of all 3 samples was also generated. The inferred transcript sets were functionally annotated and compared globally and also on subsets of secreted proteins and putative allergen families. While intermediary metabolism appeared to be typical for nematodes in the 3 evaluated taxa, their transcriptomes present strong levels of differential expression and enrichment, mainly of transcripts related to metabolic pathways and gene ontologies associated to energy metabolism and other pathways, with significant presence of excreted/secreted proteins, most of them allergens. The allergome of the 2 species and their hybrids has also been thoroughly studied; at least 74 different allergen families were identified in the transcriptomes. CONCLUSIONS: A. simplex s.s., A. pegreffi and their hybrids differ in gene expression patterns in the L3 stage. Strong parent-of-origin effects were observed: A. pegreffi alleles dominate in the expression patterns of hybrids albeit the latter, and A. pegreffii also display significant differences indicating that hybrids are intermediate biological entities among their parental species, and thus of outstanding interest in the study of speciation in nematodes. Analyses of differential expression based on genes coding for secreted proteins suggests that co-infections presents different repertoires of released protein to the host environment. Both species and their hybrids, share more allergen genes than previously thought and are likely to induce overlapping disease responses.

Morphological and genetic characterization of Hysterothylacium Ward & Magath, 1917 (Nematoda: Raphidascarididae) larvae in horse mackerel, blue whiting and anchovy from Spanish Atlantic and Mediterranean waters.

The presence of zoonotic Hysterothylacium larvae in fish from Spanish Atlantic and Mediterranean waters, which can cause economic losses for commercial fisheries, has been reported in several studies; however, little is known about species identity in this region. The aim of this study was to identify at species level the Hysterothylacium morphotypes detected in three commonly consumed fish: horse mackerel (Trachurus trachurus), blue whiting (Micromesistius poutassou) and anchovy (Engraulis encrasicolus). Third- and fourth-stage Hysterothylacium larvae, as well as adults obtained from larval in vitro culture, were morphologically and molecularly identified by ITS1/ITS2 rDNA sequencing. Four Hysterothylacium morphotypes were detected. Genetic analysis showed that morphotypes VIII and IX were different larval stages of Hysterothylacium aduncum, which was supported by cultured adult species identification. Morphotypes III and IV were found to correspond to different developmental stages of another species of Hysterothylacium. As all larval types detected were morphologically indistinguishable from others previously reported yet showed clear genetic differences, they are referred here as new genotypes. This is the first time that ITS-sequence data of various developmental stages of the same species, including adults, have been studied and compared, providing crucial knowledge for future studies on Hysterothylacium identification and biology.

A scanning electron microscopy study of Anisakis physeteris molecularly identified: from third stage larvae from fish to fourth stage larvae obtained in vitro.

The development of the fourth larval stage (L4) of Anisakis physeteris was studied using scanning electron microscopy (SEM), comparing it with third larval stage (L3) recently obtained from the host fish, blue whiting (Micromesistius poutassou), from the western Mediterranean Sea (east coast of Spain, zone FAO 37.1.1). After molting to L4, samples of the parasite were examined at different times in order to observe their development. Following collection of the L4, a small portion was taken from the middle of the larva for molecular identification, confirming in all cases that it was A. physeteris. The anterior and posterior sections of the larvae were prepared for morphological study by SEM. The development of a row of denticles on each of the three prominent lips, almost reaching the buccal commisures, was observed in the L4. Pores of unknown function were found in the upper external part of each lip. Clearly developed cephalic papillae, amphids, and deirids were also observed in L4, while, although present in L3, these were beneath the cuticle. Phasmids were detected in L4 but not in L3. The L4 tail finished in a conical lobe with a blunt point, absent in L3. In the oldest L4, some preanal papillae were observed beneath the cuticle in males, while, in females, the vulva could be seen by light microscopy, apparently still covered by the cuticle.

Occurrence of Anisakis and Hysterothylacium larvae in commercial fish from Balearic Sea (Western Mediterranean Sea).

This study investigates the occurrence of anisakids and raphidascarids in commercial fish from Balearic Sea (Western Mediterranean). A total of 335 fish including 19 black anglerfish (Lophius budegassa), 33 white anglerfish (L. piscatorius), 129 European hake (Merluccius merluccius), 30 red mullet (Mullus barbatus), and 124 striped mullet (M. surmuletus) were examined using enzymatic digestion. A total of 948 nematode larvae were isolated (prevalence 52.53%) being the highest prevalence observed in striped mullet. Forty-six larvae were identified using molecular analyses which included PCR and sequencing of the 629-bp fragment of mitochondrial cox2 gene region. Anisakis pegreffii (80.43%), A. physeteris (8.69%), Hysterothylacium fabri (6.52%), and A. simplex (4.35%) were detected based on molecular analyses of larvae. Total nematode prevalence was positively correlated with weight, length, condition factor, and maturity stage of the host and also with fishing ground depth. Statistical differences between total nematode prevalence and geographical sector of capture were observed when fishing hauls were grouped according to the abundance of sperm whales or common bottlenose dolphins. The results also corroborate that fishing water depth may play an important role in anisakid and raphidascarid parasitization.

New Antarctic deep-sea weird leech (Hirudinida: Piscicolidae): morphological features and phylogenetic relationships.

A new fish leech Ambulobdella shandikovi n. g., n. sp. (Hirudinida: Piscicolidae), a parasite of Whitson's grenadier Macrourus whitsoni (Regan) (Macrouridae: Gadiformes) collected in the Ross Sea at depths from 1,221 to 1,433 m, is described and compared with related taxa based on morphological and molecular characters. Ambulobdella shandikovi n. sp. is characterised by prominent segmental tubercles on the venter and dorsal segmental tubercles, an uncommon appearance of its anterior sucker with ear-like edges and an inner membrane around the mouth-pore, well-developed musculature and a unique combination of features of the reproductive and digestive systems. The presence of uncommon tubercles can be attributed, in part, to temporary associations of A. shandikovi n. sp. with its fish hosts and a need for well-developed sensory and locomotory organs. A certain locomotory function of ventrolateral tubercles of A. shandikovi n. sp. is hypothesised and discussed. Further deep-sea surveys are obviously needed to shed light on the behaviour and mode of locomotion of this species.

Myxosporean parasites of Ceratomyxa merlangi and Myxidium gadi in whiting Merlangius merlangus: a comparative epizootiological analysis based on samples from two localities off southern and northern coasts of the Black Sea.

Totally 690 and 423 whiting Merlangius merlangus samples were collected from two localities off Southern (Sinop) and Northern coasts (Balaklava Bay) of the Black Sea, respectively, and examined for myxosporeans in the period from May 2011 to March 2014. Ceratomyxa merlangi and Myxidium gadi were the only myxosporean parasites identified in the content of gall bladder and their infection indices of prevalence (%) and intensity were calculated for length classes and sex of fish as well as for the seasons at both sampling localities. Overall infection prevalence of C. merlangi was 22.6% in Sinop and 27.9% in Balaklava samples while those values of M. gadi were 18.4% and 28.6% in Sinop and Balaklava samples, respectively. Both parasite species were also found to be co-existed in whiting samples. This is the first comprehensive epizootiological study yielded comparable data on C. merlangi and M. gadi infections in whiting in the southern and northern coasts of the Black Sea.

Ultrastructural characterization of Pleistophora macrozoarcidis Nigerelli 1946 (Microsporidia) infecting the ocean pout Macrozoarces americanus (Perciformes, Zoarcidae) from the gulf of Maine, MA, USA.

Pleistophora macrozoarcidis a microsporidian parasite infecting the muscle tissue of the ocean pout Macrozoarces americanus collected from the Gulf of Maine of the Atlantic Ocean, MA, USA, was morphologically described on the basis of ultrastructural features. Infection was detected as opaque white or rusty brown lesions scattered throughout the musculature of the fish mainly in the region anterior to anus. Transmission electron microscopy showed that in individual parasitized muscle cells, the infection progresses within parasite formed vesicles which are in direct contact with muscle cell elements. The earliest observed parasitic stages are the globular multinucleated proliferative cells or plasmodia limited by a highly tortuous plasmalemma with intervesicular finger-like digitations projecting into the parasite cytoplasm. These cells divided through the invagination of the plasmalemma and the amorphous coat producing daughter-cells. Fine electron-dense secretion is deposited on the plasmalemma that causes its thickening which is a sign of commencement of the sporogonic phase. This phase is carried out by cytokinesis of the sporonts and results in the formation of sporoblasts and finally spores. Mature spore has a thin electron-dense exospore, a thick electron-lucent endospore, and the plasma membrane which encloses the spore contents. A single nucleus is centrally located with the posterior region containing a posterior vacuole. The majority of spores have 7-13 coils in 1-2 rows, and a small group of spores had about 23 coils forming two rows. Events of polar filament extrusion for penetration of uninfected cells were studied. The polaroplast membranes were expanded and occupy most of the length of the spore. The coils are dislocated from the sides of the spore to throughout the entire sporoplasm. The polar filament everts and extrudes through the polar cap with a sufficient force to pierce adjacent sporophorous vesicle walls. After eversion, the polar filament is referred to as a polar tubule, as it forms a tube through which the sporoplasm travels. It pierces anything in its path and deposits the sporoplasm at a new location to begin another infective cycle.

A new species of Hamaticolax Ho & Lin, 2006 (Copepoda: Bomolochidae) from deep water macrourids in the Mediterranean.

Hamaticolax resupinus n. sp. is described from specimens collected from the gill cavities of Coelorinchus mediterraneus Iwamoto & Ungaro and Coryphaenoides mediterraneus (Giglioli) (Gadiformes: Macrouridae) caught in the Western Mediterranean Sea at depths between 1,236 and 1,626 m. Hamaticolax resupinus n. sp. closely resembles H. maleus Oldewage, 1994, but differs from the latter by its smaller body size and in having a genital double-somite in the female that is markedly wider than the free abdominal somites and has strongly convex lateral margins. The new species is only the second bomolochid found on a macrourid host and is the first from depths in excess of 1,200 m. Hamaticolax resupinus n. sp. also represents the first parasitic copepod recorded from Coe. mediterraneus and only the third one from Cor. mediterraneus worldwide.

Morphological and molecular characterisation of Aporocotyle margolisi Smith, 1967 (Digenea: Aporocotylidae) from the North Pacific hake Merluccius productus (Ayres) (Gadiformes: Merlucciidae) off Oregon, USA.

Aporocotylid blood flukes conspecific with Aporocotyle margolisi Smith, 1967 were collected from the bulbus arteriosus of the North Pacific hake Merluccius productus (Ayres). This study revisits the morphology of A. margolisi, including drawings, measurements and scanning electron microscopy images, and provides for the first time molecular data for the large subunit of the ribosomal RNA (28S rDNA) and the mitochondrial cytochrome c oxidase subunit 1 (cox1) genes for this species. A 28S rDNA phylogenetic study of A. margolisi, and all available Aporocotyle spp., was also performed. The distribution range of A. margolisi is extended to the Pacific coast of the USA. We provide a morphological comparison of Aporocotyle spp. from the Pacific coast in North America as well as other Aporocotyle spp. infecting hake. Comparisons with the original description revealed that the new specimens of A. margolisi were considerably larger with respect to all morphological features, except for shorter spines. Molecular results showed a close relationship between A. margolisi and A. argentinensis Smith, 1969 from the Argentine hake Merluccius hubbsi Marini. The phylogenetic relationships of Aporocotyle spp. point to a possible co-speciation of hakes species and these blood fluke parasites.

Myxozoans as biological tags for stock identification of the Argentine hake, Merluccius hubbsi (Gadiformes: Merlucciidae).

Myxozoans have been successfully used as tags for fish stock identification around the world. However, few studies using myxozoan tags have been carried out in the Southern Atlantic, a region with complex oceanography that constitutes a potentially suitable scenario for testing the utility of myxozoans as indicators. Its usefulness was tested using six samples of Merluccius hubbsi in two different regions of the Argentine Sea. Generalized linear models were performed to assess the effects of fish size and sex, and year and region of capture and selected using the Information Theoretic approach. Three myxozoan species were recorded: Kudoa rosenbuschi, Myxoproteus meridionalis and Fabespora sp. Results of modelling species individually showed differential capabilities for detecting geographical population structure at different spatial scales, with K. rosenbuschi and Fabespora sp. allowing the discrimination of northern and southern stocks, but Fabespora sp. also as a promissory indicator of intrapopulation sub-structure due to different migratory routes during non-reproductive periods. This work confirms that myxozoans offer a set of suitable markers at different spatial scales, which can be selected individually or in any combination, depending on the geographical extent of the study, constituting tools adaptable to the objectives of further research on fish population structure.

Levels of infection and seasonal occurrence of Gyrodactylus alviga (Monogenea: Gyrodactylidae) on the whiting Merlangius merlangus off the Turkish and Russian coasts of the Black Sea.

Gyrodactylus alviga is a generalist species reported in many Black Sea fish species, but whiting is known to be its main host. It is the only monogenean parasite that has been reported so far on the skin, fins and gills of whiting Merlangius merlangus in the Black Sea. A total of 690 fish from Turkey and 423 fish from Russia were examined to detect parasites. Infection indices of prevalence, mean intensity and mean abundance values were calculated for length classes and sex of fish, as well as for the seasons. There is a gradual increase in infection indices in relation with increasing host length classes and there are higher infection indices on female fish. Furthermore, despite lower prevalence indices in spring and winter, higher parasite intensity values were determined in these seasons. It is concluded that larger fish provided more space and food for G. alviga, and female fish were more parasitized as they were more active in searching for food, which enables the parasite to switch between hosts. In addition, winter and spring were more appropriate for reproduction of G. alviga.

Removal of Anisakis simplex allergens from infected fish during the washing step of surimi production.

BACKGOUND: The washing operation of fish muscle is one of the key steps in the production of surimi. The aim of this study was to assess in parasitised minced fish the effect of the washing steps on the allergen removal of Anisakis simplex and on protein yield during surimi processing. Experimentally infected hake (Merluccius merluccius) (50 Anisakis simplex s.s L3 larvae per 100 g of muscle) underwent three successive washing steps with water, phosphate buffer (20 mmol L(-1) ), sodium bicarbonate (60 mmol L(-1) ), or sodium hypochlorite (0.27 mmol L(-1) ) in the surimi processing (4 kg muscle, 1:4 w/v for each solution). Total protein concentration and A. simplex antigens and allergens were evaluated in each waste fraction. RESULTS: The highest removal of Ani s 4 and A. simplex antigens was achieved by using phosphate buffer, together with a good protein yield in the raw surimi. Decrease of the concentration of allergens and antigens as a function of the washing steps rendered a linear trend (R(2) = 0.95 and 0.98 for Ani s 4 and A. simplex antigens, respectively). CONCLUSION: The conditions for an optimal removal of Anisakis allergens can be established and calculated as a function of the washing steps. This approach opens a line to utilise parasitised fish in a safer way. © 2014 Society of Chemical Industry.