Hepatic immune response of Coilia nasus infected with Anisakidae during ovarian development.

Anisakidae parasitism is a prevalent disease in wild populations of Coilia nasus, and can result in a significant loss of germplasm resources. To elucidate the immune response mechanism of C. nasus livers to Anisakidae infection, we collected and analysed 18 parasitic and 18 non-parasitic livers at gonadal developmental stages II, III, and V using histopathology, molecular biology and transcriptome methods. The hepatic portal area of the parasitic group exhibited an increase in the fibrous stroma and thickened hepatic arteries with positive Ly-6G staining, indicating inflammation and immune responses in the liver. Hepatocyte cytokine levels and the expression of liver function-related genes indicated that fish livers responded similarly to Anisakidae parasitism across different gonadal developmental stages. Oxidative stress indices showed more intense changes in stage II samples, whereas gene expression levels of Nrf2 and C3 were significantly increased in parasitised livers during stage III and V. Liver transcriptome sequencing identified 2575 differentially expressed genes between the parasitic and non-parasitic groups at the three gonadal developmental stages. KEGG pathway analysis showed that natural killer cell-mediated cytotoxicity, the NOD-like receptor signaling pathway, neutrophil extracellular trap formation, and other immune pathways were significantly enriched. Expression patterns varied across developmental stages, suggesting that innate immunity was primarily responsible for the liver immune response to Anisakidae infection during C. nasus migration, possibly related to water temperature changes or shifts in the gonadal developmental stage. In summary, this study investigated the immune response of C. nasus to Anisakidae parasitism under natural conditions, focusing on reproductive aspects and environmental changes, thereby establishing a foundation for elucidating the molecular mechanisms underlying the immune response of Anisakidae in C. nasus.

Re-evaluation of certain aspects of the EFSA Scientific Opinion of April 2010 on risk assessment of parasites in fishery products, based on new scientific data. Part 1: ToRs1-3.

Surveillance data published since 2010, although limited, showed that there is no evidence of zoonotic parasite infection in market quality Atlantic salmon, marine rainbow trout, gilthead seabream, turbot, meagre, Atlantic halibut, common carp and European catfish. No studies were found for greater amberjack, brown trout, African catfish, European eel and pikeperch. Anisakis pegreffii, A. simplex (s. s.) and Cryptocotyle lingua were found in European seabass, Atlantic bluefin tuna and/or cod, and Pseudamphistomum truncatum and Paracoenogonimus ovatus in tench, produced in open offshore cages or flow-through ponds or tanks. It is almost certain that fish produced in closed recirculating aquaculture systems (RAS) or flow-through facilities with filtered water intake and exclusively fed heat-treated feed are free of zoonotic parasites. Since the last EFSA opinion, the UV-press and artificial digestion methods have been developed into ISO standards to detect parasites in fish, while new UV-scanning, optical, molecular and OMICs technologies and methodologies have been developed for the detection, visualisation, isolation and/or identification of zoonotic parasites in fish. Freezing and heating continue to be the most efficient methods to kill parasites in fishery products. High-pressure processing may be suitable for some specific products. Pulsed electric field is a promising technology although further development is needed. Ultrasound treatments were not effective. Traditional dry salting of anchovies successfully inactivated Anisakis. Studies on other traditional processes - air-drying and double salting (brine salting plus dry salting) - suggest that anisakids are successfully inactivated, but more data covering these and other parasites in more fish species and products is required to determine if these processes are always effective. Marinade combinations with anchovies have not effectively inactivated anisakids. Natural products, essential oils and plant extracts, may kill parasites but safety and organoleptic data are lacking. Advanced processing techniques for intelligent gutting and trimming are being developed to remove parasites from fish.

First record of Pseudoterranova decipiens in sprat (Sprattus sprattus) from the Baltic Sea.

A dispersion of Anisakidae nematodes (particularly Contracaecum osculatum) among marine organisms in the Baltic Sea has been reported over the last decade. This is in line with an increase in the number of grey seal that act as final host for Contracaecum osculatum and Pseudoterranova sp., and are thus indispensable for the completion of their life cycles. Most attention has been paid to zoonotic nematode species, like Pseudoterranova sp., which have been noted in commercially important fish in the area. Little is known about the spread and transmission of Pseudoterranova sp. in the Baltic Sea. The aim of this study was to investigate whether sprat may play a role as a transport host for this Anisakidae. Samples were collected in three areas of the southern Baltic Sea (south and east of Bornholm, Slupsk Farrow and the Gulf of Gdansk) during a research cruise in August 2019. A visual inspection of the viscera of 556 sprats was conducted. Parasites were identified using anatomomorphological and molecular methods. Nematodes were recorded only in sprat caught southeast of Bornholm (prevalence 2.7%; intensity of infection 1-4; abundance 0.05). Molecular identification revealed the presence of Pseudoterranova decipiens. This is the first report of P. decipiens in sprat from the Baltic Sea. Sprat is likely a transmitter of P. decipiens in the Baltic Sea food web.

Identification of Contracaecum rudolphii (Nematoda: Anisakidae) in Great Cormorants Phalacrocorax carbo sinensis (Blumenbach, 1978) from Southern Italy.

In this study, four dead great cormorant Phalacrocorax carbo sinensis (Blumenbach, 1978) specimens, collected from the coasts and lakes of Southern Italy, were examined by necropsy for the detection of Contraceacum sp. The adults and larvae found were subjected to morphological analysis and molecular identification by PCR-RFLP. A total of 181 Contracaecum specimens were detected in all of the four great cormorants examined (prevalence = 100%), showing an intensity of infestation between nine and ninety-two. A co-infestation by adult and larval forms of Contracaecum rudolphii was found only in one of the great cormorants examined. Following molecular investigations, 48 specimens of C. rudolphii A and 38 specimens of C. rudolphii B were detected, revealing co-infestation solely for the great cormorant from Leporano Bay (Southern Italy). Our results showed an opposite ratio between C. rudolphii A and C. rudolphii B in Pantelleria and in Salso Lake (Southern Italy) compared to what was reported in the literature, probably due to migratory stopovers and the ecology of the infested fish species, confirming the role of Contracaecum nematodes as ecological tags of their hosts.

Metazoan endoparasites of Hoplias malabaricus (Bloch, 1794) (Actinopterygii: Erythrinidae) from upper and middle São Francisco river basin, Minas Gerais State, Brazil.

The endoparasitic fauna of Hoplias malabaricus (which is a species of paramount importance in the fishing and human food sectors) from Três Marias reservoir, São Francisco river, and from eleven marginal lagoons in the upper and middle São Francisco river basin, Brazil, was herein recorded for the first time. In total, 13 endoparasite species belonging to ten different families were found in 147 analyzed H. malabaricus specimens. The identified taxa comprised individuals belonging to phyla Apicomplexa-Calyptosporidae-Caplyptospora sp. (oocysts); Platyhelminthes-Trematoda-Diplostomidae (metacercariae)-Austrodiplostomum sp. and Sphincterodiplostomum musculosum, Clinostomidae (metacercariae)-Clinostomum sp., Gorgoderidae (adults)-Phyllodistomum spatula, and Eucestoda-Proteocephalidae gen. sp. (plerocercoids larvae); and Nematoda-Anisakidae (larvae)-Contracaecum sp. Types 1 and 2 and Hysterothylacium sp., Gnathostomatidae (larvae)-Spiroxys sp., Camallanidae (juveniles/adults)-Procamallanus (Spirocamallanus) inopinatus, Guyanemidae (juveniles/adults)-Guyanema baudi, and Cystidicolidae (juveniles/adults)-Cystidicoloides fischeri. Proteocephalidae gen. sp. and Contracaecum sp. Type 1 were the species presenting expressive parasitic indexes in the reservoir, in the river, and in nine of the eleven lagoons. Cystidicoloides fischeri was recorded for the first time in H. malabaricus. Guyanema baudi and S. musculosum had their geographic distribution expanded to São Francisco river basin.

Faunal change over 3 decades to reveal Rotundocollarette capoori n.gen., n.sp., a unique anisakid from Johnius dussumieri from the Central west coast of India at Goa.

A unique anisakid roundworm, Rotundocollarette capoori n. gen., n.sp. infesting Johnius dussumieri from the Central West coast of India at Goa has been described. The worms are typically characterized by a cephalic collarette, a muscular collarette in post-caudal zone, intestinal caecum and a ventricular appendix. 3 pores, a part of cephalic infrastructure-a dorsal oral aperture, a dorso-laterally placed genital pore, and a sub-ventral excretory pore, at the junction of the bases of the 2 ventro-lateral lips, were typically unique as revealed by SEM studies. A pair of large squarish teeth flanked each of these three pores in the cephalic region, with 3-5 denticles in the median group, at the junction of the two arms of V-shaped structure, that joined 26-29 denticles on each arm of this V-shaped formation, running parallel and crossed past dentigerous ridges on each lip. In addition, a set of 49-50 medially organized denticles occupying the dorsal arm of V-shaped formation, in the medial space, were a conspicuous configuration, besides 70 min triangular denticles at the junction of sub-ventral lips. A spined mucron at the tip of tail. The key provided by Moravec and Justine (Parasite 27:58, 2020. 10.1051/parasite/2020053) to segregate Porrocaecum-like nematode genera (Porrocaecum, Pseudoterranova, Pulchrascaris, Euterranova, and Neoterranova) has been revised to include the newly proposed genus, Rotundocollarette capoori n.gen., n.sp. The significance of the emerging nematode as an environmental bioindicator has been specified.

Large-scale genetic investigation of nematode diversity and their phylogenetic patterns in New Zealand's marine animals.

Nematodes constitute one of the most speciose metazoan groups on earth, and a significant proportion of them have parasitic life styles. Zooparasitic nematodes have zoonotic, commercial and ecological significance within natural systems. Due to their generally small size and hidden nature within their hosts, and the fact that species discrimination using traditional morphological characteristics is often challenging, their biodiversity is not well known, especially within marine ecosystems. For instance, the majority of New Zealand's marine animals have never been the subject of nematode studies, and many currently known nematodes in New Zealand await confirmation of their species identity with modern taxonomic techniques. In this study, we present the results of an extensive biodiversity survey and phylogenetic analyses of parasitic nematodes infecting New Zealand's marine animals. We used genetic data to differentiate nematodes to the lowest taxonomic level possible and present phylogenies of the dominant clades to illustrate their genetic diversity in New Zealand. Our findings reveal a high diversity of parasitic nematodes (23 taxa) infecting New Zealand's marine animals (62 of 94 free-living animal species investigated). The novel data collected here provide a solid baseline for future assessments of change in diversity and distribution of parasitic nematodes.

The effects of Contracaecum osculatum larvae on the growth of Atlantic cod (Gadus morhua).

Atlantic cod (Gadus morhua) from the Eastern Baltic stock have decreased in numbers and condition since the 1990's. Among several causes, an increased prevalence and intensity of the nematode Contracaecum osculatum has been discussed. This increase has been attributed to a population increase of the parasites final host, the grey seal (Halichoerus grypus). Other studies have looked at the role of Contracaecum osculatum on cod growth and condition on recently caught cod, or done short term experimental studies in lab. This study instead investigated the importance of Contracaecum osculatum for cod growth in a sea pen based experiment, where cod were kept and fed in order to monitor growth. The results show that a higher density (number of nematodes per gram liver) decreases cod growth potential. If the number of nematodes exceeded 8 per gram liver cod did not grow in length, even when given generous amounts of food. Accounting for the lack of growth due to Contracaecum osculatum may improve stock assessments and increase the possibility to reach management targets.

Contracaecum nematode parasites in hillstream loaches of the Western Ghats, India.

Nematode parasites of the family Anisakidae infect definitive hosts, such as fish-eating birds and mammals, through primary intermediate hosts like copepods and secondary intermediate hosts like fishes. However, consumption of raw or undercooked fish can lead to nematode infection called anisakidosis in humans. We observed the presence of nematode infection in hillstream loaches of families Cobitidae and Nemacheilidae available for human consumption in the local markets in the northern parts of Western Ghats, India. Scanning electron micrograph and genetic identification employing mitochondrial cytochrome oxidase subunit II, identified the nematode to the genus Contracaecum. Histology of infected host revealed the presence of the parasite in muscles. Antioxidant enzyme analysis of host liver suggested that infection leads to oxidative stress in the fish. We suspect that a gradual increase in parasite infection of the loaches in the last decade could be attributed to various anthropogenic stressors that are altering riverine habitats. Since loaches are consumed by tribal people who often prepare the fish without degutting and possibly undercooked, there is a potential threat of human infection.

Gene expression and allergenic potential of Pseudoterranova bulbosa L3 from different infection sites in North Atlantic cod (Gadus morhua).

The recent advances in molecular methods and data processing have facilitated research on anisakid nematodes. While most research efforts were made regarding the genus Anisakis, since this genus is held responsible for the majority of reported clinical signs, there is still a demand for data on the genus Pseudoterranova. Several case studies of severe invasive anisakidosis affecting various organs caused by species of the P. decipiens complex have been described. To better understand the way these parasites might infest their fish host, we examined whether parasite location within the fish host affects gene expression. A de novo assembly of the transcriptome of Pseudoterranova bulbosa, isolated from North Atlantic cod, was analysed for patterns of differential gene expression between samples taken from liver and viscera. We additionally searched for homologs to known nematode allergens, to give a first estimate of the potential allergenicity of P. bulbosa. There was a subtle difference in the gene expression of samples taken from liver and viscera. Seventy genes were differentially expressed, 32 genes were upregulated in parasites isolated from liver and 38 genes were upregulated in parasites from viscera. Homologs of five nematode allergens were identified among the genes expressed by P. bulbosa. Our transcriptome of P. bulbosa will be a valuable resource for further meta-analyses and resequencing projects.

Drug efficacy on zoonotic nematodes of the Anisakidae family: new metabolic data.

In the Anisakidae family, there are nematodes, most of which are parasitic for important commercial fish species. Both public health risks and socio-economic problems are attributed to these parasites. Despite these concerns, knowledge of the metabolism of these parasites remains unknown. Therefore, the main objective of this study was to investigate the receptors of drugs and oxidative metabolic status of two Anisakidae species, Pseudoterranova decipiens (s. s.) and Contracaecum osculatum (s. s.), under the influence of anthelminthic drugs, ivermectin (IVM) and pyrantel (PYR), at different concentrations: 1.56, 3.125 and 6.25 µg mL−1 of culture medium for 3, 6, 9, 12 and 72 h. The mRNA expressions of the γ-aminobutyric acid receptor, acetylcholine receptor subunits, adenosine triphosphate-binding cassette transporters and antioxidative enzymes were determined. The total antioxidant capacity and glutathione S-transferase activity were also examined. To the best of the authors' knowledge, this is the first time that IVM and PYR have been tested against these parasitic nematodes.

Unraveling the microbiota of the fish parasite Pseudoterranova decipiens in codfish (Gadus morhua) reveals a fish-related bacterial community.

Anisakidae, mainly represented by the species Anisakis simplex and Pseudoterranova decipiens, are one of the most commonly zoonotic nematodes present in marine fish species. Apart from public health risks directly linked to the parasite itself, little is known on the effects of the migrating nematodes on the hygienic quality of the fish fillet due to bacteria it carries. In the present study, the cultivated bacterial community on and in individual P. decipiens larvae deriving from codfish is reported. Four isolation media were included and evaluated to increase the bacterial diversity isolated, and identification of the bacterial growth was performed by a combination of Matrix-Assisted Laser Desorption Ionization Time-Of-Flight mass spectrometry and 16S rRNA gene sequencing. Results revealed that the microbiota of P. decipiens larvae comprises both potential spoilage bacteria and human opportunistic pathogens, and that a combined isolation on the general isolation medium tryptone soy agar and a medium supplemented with artificial seawater resulted in the highest bacterial recovery in terms of diversity and enumeration. Dissimilarity analysis also revealed similar, though unique, bacterial communities between nematodes originating from the same fish suggesting that anisakid microbiota compositions are reflections of the microbial assemblages in the fish host as an individual, and that the gut microbiome is diverse within gadoid fish species originating from the same geographical habitat. Future research should, based on the results in the present study, further elaborate on the comparison of the bacterial communities of both the larva and the codfish from which it was isolated, and, explore the extrapolation potential towards other fish and nematode species. Also, the actual degree of risk beyond the simple presence of the parasite due to carriage of opportunistic bacteria should be examined, as well as the nematode's true effect on spoilage.

Endoparasite diversity and liver alterations in Hoplerythrinus unitaeniatus and Cichlasoma bimaculatum in a quilombola area in Maranhão, Brazil / Diversidade endoparasitária e alterações hepáticas em Hoplerythrinus unitaeniatus e Cichlasoma bimaculatum de área quilombola maranhense, Brasil

Our aim was to assess endoparasite diversity and liver alterations in Hoplerythrinus unitaeniatus (jeju) and Cichlasoma bimaculatum (acará preto) in a quilombola area in Maranhão, Brazil. For this, 21 H. unitaeniatus and 21 C. bimaculatum were caught in a natural environment and transported to a laboratory. After these had been euthanized, endoparasites were collected and identified. Liver alterations were evaluated histological analysis based on the severity of each lesion: stage I, organ functioning not compromised; stage II, more severe lesions that impair normal functioning of the organs; and stage III, very severe and irreversible lesions. Among the fish evaluated, 71.43% H. unitaeniatus and 61.90% C. bimaculatum were parasitized. Contracaecum sp. was found in both species; while acanthocephalans, only in H. unitaeniatus. The alterations were vacuolization, nucleus in the cell periphery, deformation of the cell outline, melanomacrophage center, hyperemia, cytoplasmic degeneration and nuclear vacuolization. Through calculating a histological alteration index, it was found that 26.19% of the specimens presented lesions in stage I; 38.09% lesions in stage II and 9.52% lesions in stage III. It was concluded that there is high prevalence of Contracaecum sp. and that the liver lesions may be adaptive responses by the fish to endoparasitic infection.(AU)

Objetivou-se avaliar a diversidade de endoparasitos e alterações hepáticas em Hoplerythrinus unitaeniatus (jeju) e Cichlasoma bimaculatum (acará preto) de área quilombola maranhense, Brasil. Assim, 21 H. unitaeniatus e 21 C. bimaculatum foram capturados de ambiente alagável e transportados vivos ao laboratório. Após a eutanásia, procedeu-se à coleta, identificação de endoparasitos e avaliação de alterações hepáticas por meio de análise histológica, baseada na severidade das lesões: (i) alterações de estágio I, não comprometem o funcionamento dos órgãos; (ii) estágio II, lesões mais severas que prejudicam o funcionamento normal dos órgãos; (iii) estágio III, lesões muito severas e irreversíveis. Dos peixes avaliados, 71,43% H. unitaeniatus e 61,90% C. bimaculatum estavam parasitados. Contracaecum sp. foi encontrado nas duas espécies e acantocéfalos apenas em H. unitaeniatus. As alterações hepáticas foram vacuolização, núcleo na periferia das células, deformação do contorno celular, centro de melanomacrófagos, hiperemia, degeneração citoplasmática e vacuolização nuclear. Com o cálculo do índice das alterações, constatou-se que 26,19% dos exemplares apresentaram alterações de estágio I; 38,09% de estágio II e 9,52% de estágio III com a constatação de larvas de Contracaecum sp. encistadas no fígado. Conclui-se que existe alta prevalência de Contracaecum sp., e lesões hepáticas podem ser respostas adaptativas dos peixes à infecção endoparasitária.(AU)

Anisakis pegreffii Larvae in Sphyraena viridensis and Description of Granulomatous Lesions.

The aim of the present study was to describe gastric granuloma caused by Anisakis pegreffii in Sphyraena viridensis caught in the central Mediterranean Sea. Sixty-eight S. viridensis specimens were collected from different fish markets on the east coast of Sicily. Coelomic organs were observed both macroscopically and with the aid of stereomicroscope. Parasite specimens and lesioned tissues were collected for identification, histological and molecular analyses. Twelve specimens (p = 17.6%) were positive for the presence of nematode larvae, morphologically identified as larvae of Anisakis sp., with values of mean abundance and mean intensity of 0.9 and 4.8, respectively. One large female specimen showed massive parasite infection associated with nodular lesions of the gastric wall. By histology, several nematode larvae encysted through the gastric wall were found. The parasite bodies were surrounded by a granulomatous reaction made up of macrophages, epithelioid cells, some lymphocytes and an external connective sheet. Molecular analysis of 18S rRNA and cox2 genes from Anisakis sp. collected larvae, identified them as A. pegreffii. The lesions here described, though macroscopically superimposable on human eosinophilic granuloma, microscopically showed significant differences in the inflammatory cells involved and in the type of immune reaction.

Occurrence and abundance of zoonotic nematodes in snapper Chrysophrys auratus, a popular table fish from Australian and New Zealand waters.

In Australia and New Zealand (NZ), snapper Chrysophrys auratus is known for delicate mild flavoured flesh and is a favoured species to serve raw as sashimi or in sushi. The diet of snapper includes a variety of intermediate hosts of larval nematodes, and as a result, snapper has potential to become highly infected with zoonotic/non-zoonotic nematodes. The aims of this study were to survey nematodes in snapper from Australia and New Zealand waters and to identify nematode species using combined morphological and molecular methods. The zoonotic potential of nematodes identified in this study are discussed. A total of 112 snapper were purchased from the Sydney fish market, New South Wales, Australia. Fish were dissected and only the visceral content and digestive tract were examined for nematode infection. Parasites were initially identified by the microscopic method as four different types belonging to the families Anisakidae (Anisakis types I & III, and Terranova type II) and Cucullanidae (Dichelyne spp.). All Anisakidae nematodes were at infective stages. Species-level identification was actualised through sequencing of the internal transcribed spacer (ITS-1, 5.8S, ITS-2) regions. The Anisakis types I & III were confirmed as Anisakis pegreffii and A. brevispiculata, respectively of which A. pegreffii is considered globally as a zoonotic nematode. The specific identification of Terranova type II and Dichelyne spp. was not possible as no comparable sequence data were available in GenBank. The phylogenetic tree clustered Anisakis types I & III with A. pegreffii and A. brevispiculata, respectively; Terranova type II sequences as a separate clade with previously identified larval and adult Terranova and Pseudoterranova species. Based on phylogenetic analyses the present Cucullanid specimens were assigned herein as Dichelyne cf. pleuronectidis, and an unknown species Dichelyne sp. 1. This study represents the first host record globally for zoonotic Anisakid nematodes in this popularly consumed table fish and a new region record for D. cf. pleuronectidis and Dichelyne sp. 1. Further investigation is required, using more comprehensive parasite detection and recovery methods, to assess the health risk these nematodes may pose to human and fish health in Australia/NZ.

Increasing intensities of Anisakis simplex (Rudolphi, 1809 det. Krabbe, 1878) larvae with weight and sea age in returning adult Atlantic salmon, Salmo salar L., of coastal waters of Norway.

Ninety wild Atlantic salmon, Salmo salar L., (1.5-10.3 kg) were caught in the Namsen Fjord near the mouth of River Namsen, mid-Norway, and examined for the presence and distribution of Anisakis simplex (Rudolphi, 1809 det. Krabbe, 1878) larvae by digestion of the viscera and muscles in a pepsin/HCl solution. All salmon were migrating spawners after 1-4 years of feeding in the Atlantic Ocean. All 90 Atlantic salmon had A. simplex larvae in the viscera, and all, except two, had A. simplex larvae in the musculature. The number of A. simplex larvae in each fish varied between 3 and 181, and the total mean number of nematode larvae was 44.5. The intensity of A. simplex larvae was positively correlated with increasing weight and sea age of the host. However, the proportion of larvae in the muscle fillets decreased with increasing host weight and sea age. Atlantic salmon females had more A. simplex larvae than males. In all the fish examined, 70.2% of the A. simplex larvae were found in the viscera and 29.8% in the musculature. The majority (93%) of the larvae in the musculature occurred in the hypaxial sections anterior to the anus. As A. simplex larvae commonly occur in the musculature of wild Atlantic salmon, consumption of unfrozen, raw or semi-raw musculature represents a risk for humans developing anisakiasis.

New host record and geographical distribution of Nematoda parasitizing Hypophthalmus marginatus Valenciennes (Siluriformes) from the Tocantins River, Brazil

Hypophthalmus marginatus is among the most commercialized fish from the Amazon region, with fish fillets exported to Southeastern Brazil and other countries. In the present study, the H. marginatusanalyzed were parasitized by third-stage larvae of Hysterothylaciumsp. and Anisakissp. These nematodes are being reported for the first time in H. marginatus collected from the Tocantins River, representing a new host and geographical records, and expanding knowledge of the hosts of these nematodes in the Neotropical Region. A brief description with original measurements is presented.

Genera and Species of the Anisakidae Family and Their Geographical Distribution.

Nematodes of the Anisakidae family have the ability to infest a wide variety of aquatic hosts during the development of their larval stages, mainly marine mammals, aquatic birds, such as pelicans, and freshwater fish, such crucian carp, these being the hosts where the life cycle is completed. The participation of intermediate hosts such as cephalopods, shrimp, crustaceans and marine fish, is an important part of this cycle. Due to morphological changes and updates to the genetic information of the different members of the family, the purpose of this review was to carry out a bibliographic search of the genus and species of the Anisakidae family identified by molecular tests, as well as the geographical area in which they were collected. The Anisakidae family is made up of eight different parasitic genera and 46 different species. Those of clinical importance to human health are highlighted: Anisakis pegreffi, A. simplexsensu stricto, Contracaecumosculatum, Pseudoterranova azarazi, P. cattani, P. decipiens and P. krabbei. The geographical distribution of these genera and species is located mainly in the European continent, Asia and South America, as well as in North and Central America and Australia. Based on the information collected from the Anisakidae family, it was determined that the geographical distribution is affected by different environmental factors, the host and the ability of the parasite itself to adapt. Its ability to adapt to the human organism has led to it being considered as a zoonotic agent. The disease in humans manifests nonspecifically, however the consumption of raw or semi-raw seafood is crucial information to link the presentation of the parasite with the disease. The use of morphological and molecular tests is of utmost importance for the correct diagnosis of the genus and species of the Anisakidae family.

[New eating habits, new parasitic risks: The example of fish]. / De nouvelles habitudes alimentaires, de nouveaux risques parasitaires : l'exemple du poisson.

INTRODUCTION: In the past decades, the massive explosion of "Japanese" restaurants serving raw fish popularised new culinary habits in France. At the same time, consumers have made a habit of preparing raw or pickled fish dishes themselves at home. As a result, the identification of live parasitic worm larvae in raw fish flesh is common and a source of concern for professionals or amateur cooks. Sometimes, these worms are spit out or removed after fibroscopy in patients developing severe epigastric pain quickly after eating raw fish. This paper is aiming at having a quick review of the main parasites transmitted to humans by eating raw fish in France. METHODS: This article is based on the personal experience of the authors, on references preferentially from the French literature and on the results of the Fish Parasites (ANR) research program. RESULTS: From 2011 to 2014, Fish-Parasites (ANR) assessed the prevalence of parasitism in sea and freshwater fish belonging to 29 species. About 57% of sea fish were parasitised by Anisakidae. Larvae of Dibothriocephalus latus were found in pike, perch, and burbot in Lake Geneva but in none of the fish examined from Annecy or Le Bourget lakes. Concerning human anisakidosis, a retrospective survey was carried out in the years 2010 to 2014 among all medical parasitology laboratories from university hospitals in France. Thirty-seven cases of anisakidosis have been reported, including 18 cases of allergic anisakidosis. Six additional cases of severe Anisakidae allergy were reported to the National Allergovigilance Network over the same period. CONCLUSIONS: Despite the increase in consumption of raw fish, and compared to previous studies, cases of anisakidosis are decreasing, but their allergenic potential is increasing. The incidence of dibothriocephalosis, after some trend of emergence on the shores of Lake Geneva some 20 years ago, is currently decreasing, but sporadic cases of importation are still reported. Actions with professionals (investigation, providing of information) and research programs on management of parasitic risk control are being pursued and have resulted in an update of the technical instruction of the French General Directorate for Food on the control of parasitic risk in fish.

Proteomic Profiling Reveals New Insights into the Allergomes of Anisakis simplex, Pseudoterranova decipiens, and Contracaecum osculatum.

Anisakis simplex, Pseudoterranova decipiens, and Contracaecum osculatum third-stage larvae (L3) are fish-borne nematodes that can cause human anisakidosis. Although A. simplex is a known source of allergens, knowledge about the allergic potential of P. decipiens and C. osculatum is limited. Therefore, we performed comparative proteomic profiling of A. simplex, P. decipiens, and C. osculatum L3 larvae using liquid chromatography-tandem mass spectrometry. In total, 645, 397, and 261 proteins were detected in A. simplex, P. decipiens, and C. osculatum L3 larvae, respectively. Western blot analysis confirmed the cross-reactivity of anti-A. simplex immunoglobulin (Ig)G antibodies with protein extracts from P. decipiens and C. osculatum L3 larvae. The identified proteins of the Anisakidae proteomes were characterized by label-free quantification and functional analysis, and proteins involved in many essential biological mechanisms, such as parasite survival, were identified. In the proteome of A. simplex 14, the following allergens were identified: Ani s 1, Ani s 2 (2 isomers), Ani s 3 (2 isomers), Ani s 4, Ani s 8, Ani s 9, Ani s 10, Ani s 11-like, Ani s 13, Ani s fructose 1,6-bisphosphatase, Ani s phosphatidylethanolamine-binding protein (PEPB), and Thu a 3.0101. The following 8 allergens were detected in P. decipiens: Ani s 2, Ani s 3 (2 isomers), Ani s 5, Ani s 8, Ani s 9, Ani s PEPB, and Ani s troponin. In C. osculatum 4, the following allergens were identified: Ani s 2, Ani s 5, Ani s 13, and Asc l 3. Furthermore, 28 probable allergens were predicted in A. simplex and P. decipiens, whereas in C. osculatum, 25 possible allergens were identified. Among the putative allergens, heat shock proteins were most frequently detected, followed by paramyosin, peptidyl-prolyl cis-trans isomerase, enolase, and tropomyosin. We provide a new proteomic data set that could be beneficial for the discovery of biomarkers or drug target candidates. Furthermore, our findings showed that in addition to A. simplex, P. decipiens and C. osculatum should also be considered as potential sources of allergens that could lead to IgE-mediated hypersensitivity.