Anisakiasis Annual Incidence and Causative Species, Japan, 2018-2019.

Using data from 2018-2019 health insurance claims, we estimated the average annual incidence of anisakiasis in Japan to be 19,737 cases. Molecular identification of larvae revealed that most (88.4%) patients were infected with the species Anisakis simplex sensu stricto. Further insights into the pathogenesis of various anisakiasis forms are needed.

World-wide prevalence of Anisakis larvae in fish and its relationship to human allergic anisakiasis: a systematic review.

The infective stage of Anisakidae nematodes responsible for allergic reactions in humans is found in a variety of edible fish and cephalopods. The identification of geographical regions that are high risk for infected seafood may help prevent allergic reactions in humans. Despite an abundance of published literature which has identified anisakid larvae in an array of edible seafood as well as scattered reports of human allergic anisakiasis, the relationship between the two has not been fully explored. Therefore, a systematic spatio-temporal study was conducted to determine the prevalence of Anisakis spp. in fish from January 2000 to August 2020 firstly to explore the relationship between fish infection and cases of allergic anisakiasis and secondly to use fish infection data to map potential allergic anisakiasis 'hot spots'. A systematic literature search for original English text articles was conducted through search engines, Web of Science, Scopus, PubMed, Science Direct and Google Scholar. Out of 3228 articles which describe anisakid infection in fish, 264 were used for data extraction. Of 904 articles describing allergic anisakiasis, 37 were used for data extraction. A qualitative summary of the extracted data was performed using equal interval method (ArcMap software) in order to compare the global distribution of Anisakis-infected fish. Of the 152-identified fish hosts, five families were most commonly infected with Anisakis spp. These included Lophiidae (86.9%), Trichiuridae (77.05%), Zeidae (70.9%), Merlucciidae (67.8%) and Gadidae (56.8%). The hot spot areas for allergic anisakiasis were North and northeast of Atlantic Ocean, southwest of USA, west of Mexico, south of Chile, east of Argentina, Norway, UK and west of Iceland (confidence 99%). The highest rate of allergic anisakiasis was in Portugal and Norway with the prevalence rate of 18.45-22.50%. Allergologists should consider allergic anisakiasis as a public health issue particularly in high-risk countries where high prevalences in fish have been demonstrated.

Seafood-borne parasitic diseases in Australia: are they rare or underdiagnosed?

Australia is a multicultural country surrounded by water where seafood is regularly consumed. Literature suggests that some popular edible fish sold in fish markets may be infected with parasites transmissible to humans (notably, anisakids and other helminths); however the number of reported human cases due to these parasites is low. In this article we critically review topical publications to understand whether the low number of human infection is due to lack of expertise in Australia to identify and diagnose accurately seafood-borne parasitic infections. The risk these parasites pose to humans may be underestimated due to: (i) errors or inability of diagnosing these infections, primarily due to less sensitive and specific serological tests and misidentifying parasites without a taxonomist in the diagnostic team; and (ii) medical practitioners not being aware of these parasites or not considering them in the differential diagnosis even in patients with history of regular raw or undercooked seafood consumption.

Invasive anisakiasis by the parasite Anisakis pegreffii (Nematoda: Anisakidae): diagnosis by real-time PCR hydrolysis probe system and immunoblotting assay.

BACKGROUND: Anisakiasis is a fish-borne zoonosis caused by Anisakis spp. larvae. One challenging issue in the diagnosis of anisakiasis is the molecular detection of the etiological agent even at very low quantity, such as in gastric or intestinal biopsy and granulomas. Aims of this study were: 1) to identify three new cases of invasive anisakiasis, by a species-specific Real-time PCR probe assay; 2) to detect immune response of the patients against the pathogen. METHODS: Parasite DNA was extracted from parasites removed in the three patients. The identification of larvae removed at gastric and intestinal level from two patients was first obtained by sequence analysis of mtDNA cox2 and EF1 α-1 of nDNA genes. This was not possible in the third patient, because of the very low DNA quantity obtained from a single one histological section of a surgically removed granuloma. Real-time PCR species-specific hydrolysis probe system, based on mtDNA cox2 gene, was performed on parasites tissue of the three cases. IgE, IgG4 and IgG immune response against antigens A. pegreffii by Immunoblotting assay was also studied. RESULTS: According to the mtDNA cox2 and the EF1 α - 1 nDNA sequence analysis, the larvae from stomach and intestine of two patients were assigned to A. pegreffii. The Real-time PCR primers/probe system, showed a fluorescent signal at 510 nm for A. pegreffii, in all the three cases. In Immunoblotting assay, patient CC1 showed IgE, IgG4 reactivity against Ani s 13-like and Ani s 7-like; patient CC2 revealed only IgG reactivity against Ani s 13-like and Ani s 7-like; while, the third patient showed IgE and IgG reactivity against Ani s 13-like, Ani s 7-like and Ani s 1-like. CONCLUSION: The Real-time PCR assay, a more sensitive method than direct DNA sequencing for the accurate and rapid identification of etiological agent of human anisakiasis, was successfully assessed for the first time. The study also highlights the importance to use both molecular and immunological tools in the diagnosis of human anisakiasis, in order to increase our knowledge about the pathological findings and immune response related to the infection by zoonotic species of the genus Anisakis.

Prevalence and Mean Intensity of Anisakidae Parasite in Seafood Caught in the Mediterranean Sea Focusing on Fish Species at Risk of Being Raw-consumed. A Meta Analysis and Systematic Review.

OBJECTIVE: To assess the prevalence and mean intensity of anisakids in seafood caught in the Mediterranean Sea, focusing on fish species at risk of being raw-consumed. DESIGN: Systematic review and meta-analysis of studies published from 1960-2012. STUDY SELECTION: Main criteria for the inclusion of studies were as follows: Findings of anisakid larvae, in both muscles and viscera; fish species for human consumption caught in the Mediterranean Sea; prevalence and mean intensity data for each species; and sample size equal to or more than 40 fishes. RESULTS: Twelve studies were identified. Among these, four studies considered the following three fish species that are often consumed raw or preserved lightly, or not cooked thoroughly: anchovy, pilchard, and Atlantic mackerel. DATA SYNTHESIS: All pooled analyses were based on the random-effect model. Anisakids prevalence in fish muscle was 0.64% (P < 0.0001), in viscera it was 1.34% (P < 0.0001), and overall prevalence was 0.95% (P < 0.0001). Mean intensity in muscle was 2.31 (P = 0.0083), in viscera it was 1.55 (P = 0.0174), and overall it was 1.81 (P < 0.0005). Heterogeneity indices (I(2)) were significantly high with the exception of viscera mean intensity. CONCLUSIONS: Anchovy, pilchard, and Atlantic mackerel have a low prevalence and mean intensity of anisakidae larvae in both viscera and muscles. Mean Intensity was also low.

Proteomic characterization of extracellular vesicles released by third stage larvae of the zoonotic parasite Anisakis pegreffii (Nematoda: Anisakidae).

Introduction: Anisakis pegreffii is a sibling species within the A. simplex (s.l.) complex requiring marine homeothermic (mainly cetaceans) and heterothermic (crustaceans, fish, and cephalopods) organisms to complete its life cycle. It is also a zoonotic species, able to accidentally infect humans (anisakiasis). To investigate the molecular signals involved in this host-parasite interaction and pathogenesis, the proteomic composition of the extracellular vesicles (EVs) released by the third-stage larvae (L3) of A. pegreffii, was characterized. Methods: Genetically identified L3 of A. pegreffii were maintained for 24 h at 37°C and EVs were isolated by serial centrifugation and ultracentrifugation of culture media. Proteomic analysis was performed by Shotgun Analysis. Results and discussion: EVs showed spherical shaped structure (size 65-295 nm). Proteomic results were blasted against the A. pegreffii specific transcriptomic database, and 153 unique proteins were identified. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis predicted several proteins belonging to distinct metabolic pathways. The similarity search employing selected parasitic nematodes database revealed that proteins associated with A. pegreffii EVs might be involved in parasite survival and adaptation, as well as in pathogenic processes. Further, a possible link between the A. pegreffii EVs proteins versus those of human and cetaceans' hosts, were predicted by using HPIDB database. The results, herein described, expand knowledge concerning the proteins possibly implied in the host-parasite interactions between this parasite and its natural and accidental hosts.

Methods for inactivation of seafood Anisakis larvae and prevention of human anisakiasis: a mini-review.

Anisakiasis is an arising zoonosis induced by parasitic nematodes belonging to the family Anisakidae. Anisakiasis is often caused by the ingestion of larval nematodes in uncooked or minimally processed seafood dishes, which are regularly consumed by humans. Significant potential sources of infection are raw fish (e.g., sushi and sashimi) that can be found in traditional Japanese cuisine and can be part of the culinary tradition of consumption of raw or marinated fish that is particularly diffused in European countries. During the last five decades, the global prevalence of human anisakiasis has been rising, becoming an emergent major public health problem. Thus, there is an unmet need for well-defined and cost-effective methods aimed at killing Anisakis larvae, thus reducing the incidence of anisakiasis. In this mini-review, we discuss the clinical features of anisakiasis as well as the effectiveness and mechanisms of action of the main methods employed for increasing seafood safety and killing Anisakis larvae, including freezing, heating, use of high hydrostatic pressure, salting process, pepsin digestion method and use of garlic oil.

First report of human anisakidosis in Australia.

We present the first human case of anisakidosis acquired from eating locally caught fish in Australia. A 41-year-old woman experienced gastrointestinal pain, vomiting and diarrhoea of increasing severity over 3 weeks. All symptoms resolved spontaneously after a worm was passed in her faeces. Microscopic examination showed that it was a Contracaecum species larva of the family Anisakidae. Anisakidosis should be considered in patients with gastrointestinal symptoms who have recently eaten seafood.

Differential proteolytic activity in Anisakis simplex s.s. and Anisakis pegreffii, two sibling species from the complex Anisakis simplex s.l., major etiological agents of anisakiasis.

Proteolytic activity was studied in two sibling species of Anisakis (Nematoda: Anisakidae), A. simplex s.s. and A. pegreffii, throughout their in vitro development from third larval stage (L3) from the host fish (L3-0h) to fourth larval stage (L4) obtained in culture. Proteases have a significant role in the lifecycle of the parasite and in the pathogen-host relationship. Proteolytic activity peaks were detected at pH 6.0 and 8.5. Protease activity was detected in all the developmental stages of the two species studied at both pH values. These pH values were used for assaying with specific inhibitors which permitted the determination of metalloprotease activity, and, to a lesser extent, that of serine and cysteine protease. Aspartic protease activity was only detected at pH 6.0. At this pH, L4 larvae showed higher proteolytic activity than L3 larvae in both species (p < 0.001), the majority of activity being due to metalloproteases and aspartic proteases, which could be related to nutrition, especially the latter, as occurs in invertebrates. At pH 8.5, proteolytic activity was higher in A. simplex s.s. than in A. pegreffii (p < 0.01). At this pH, the majority of activity was due to metalloproteases in all developmental phases of both species, although, in L3-0h, the activity of these proteases was significantly higher (p < 0.03) in A. simplex s.s. than in A. pegreffii. This could be related to the greater invasive capacity of the former. Serine proteases have frequently been implicated in the invasive capacity and pathogenicity of some parasites. This may be related to the significantly higher activity (p ≤ 0.05) of serine protease in all the larval stages of A. simplex studied at pH 6.0. Thus, there are interspecific differences in proteases that have been related to pathogenesis in nematodes. These differences could thus be contributing to the previously reported differences in pathogenicity between these two Anisakis species.

A national retrospective survey of anisakidosis in France (2010-2014): decreasing incidence, female predominance, and emerging allergic potential.

A retrospective survey was carried out over the years 2010-2014 among all Parasitology laboratories of University hospitals in France (ANOFEL network). The objective was to estimate the incidence of anisakidosis in France as new culinary habits such as the consumption of raw fish (sushi) or undercooked fish are increasing. A total of 37 cases of anisakidosis were notified by all French laboratories: 7 proven cases with evidence of a worm, 12 possible cases with abdominal pain after consumption of raw fish with detection of anti-Anisakis precipitins, and 18 allergic cases defined as acute manifestations after consumption of fish, associated with specific IgE for Anisakis. The median age of affected individuals was 42 years (11-69) and there was a significant predominance of women (67%). Compared with previous surveys in France, this study indicates a decrease in clinical cases of anisakidosis and illustrates the emerging allergic potential of anisakids.

TITLE: Étude nationale rétrospective de l'anisakidose en France (2010-2014) : diminution de l'incidence, prédominance féminine et augmentation des allergies. ABSTRACT: Une enquête rétrospective a été réalisée au cours des années 2010-2014 auprès des laboratoires de parasitologie des hôpitaux universitaires de France (réseau ANOFEL). L'objectif était d'estimer l'incidence de l'anisakidose en France depuis l'émergence de nouvelles habitudes culinaires comme la consommation de poissons crus (sushi) ou insuffisamment cuits. Les laboratoires français de Parasitologie ont ainsi notifié 37 cas d'anisakidose: 7 cas certains avec présence du ver, 12 cas possibles se caractérisant par des douleurs abdominales après consommation de poisson cru et la présence de précipitines anti-Anisakis et 18 cas allergiques définis comme des manifestations aiguës après consommation de poisson associées à des IgE spécifiques anti-Anisakis. L'âge médian des cas était de 42 ans (11-69) et il y avait une prédominance féminine significative (67 %). Par rapport aux enquêtes précédentes en France, cette étude indique une diminution des cas cliniques d'anisakidose mais illustre le potentiel allergique émergent des anisakidés.

Human anisakiasis in Italy: a retrospective epidemiological study over two decades.

A retrospective analysis on human anisakiasis in Italy since its first description in 1996 was performed by conducting a literature search. Inclusion criteria based on the presence of a larva and on parasite identification were applied. Epidemiological data and clinical features were analysed. Particular attention was paid to the source of infection. In total, 73 cases were included in the analysis, while 34 were excluded. Cases were reported from eight Italian regions, most frequently Abruzzo, Apulia and Latium. The parasite was detected by endoscopy (51.4%) or laparotomy (48.6%). The site of infection was intestinal (42.5%), gastric (43.8%), oesophageal (1.4%) or ectopic (12.3%). Most of the parasites (71.0%) were identified as Anisakis sp. or A. simplex (s.l.). However, when molecular methods were used (21 cases), A. pegreffii was always identified. In most of the patients (65.7%), the source of infection was raw or undercooked anchovies, followed by "anchovies or sardines" (15.1%), generic "raw seafood" (15.1%), and sardines (1.4%). In only 2 cases (2.7%), the source was not available. This is the first systematic analysis of Italian cases of anisakiasis. The main conclusions derived from the results are: i) attention should be given to the history, in particular when raw marinated anchovies, proven to be the main source of human anisakiasis in Italy, are consumed; ii) in order to assess correct epidemiological data, a confirmed and specific etiological identification should always be sought.

Human gastric hyperinfection by Anisakis simplex: A severe and unusual presentation and a brief review.

Anisakiasis is an emerging marine food-borne zoonosis resulting from the accidental ingestion of Anisakis larvae, through the consumption of raw or undercooked infected seafood products. The first case of human gastric hyperinfection by Anisakis simplex with an unusual and severe presentation, occurring in a Portuguese woman, is described in this article. Over 140 anisakid larvae were removed by gastroscopy. Massive infection is uncommon in areas where the consumption of raw fish is not part of the traditional diet, as is the case in Portugal. The increased consumption of raw seafood products is considered a health determinant in the rise in cases of anisakiasis. However, clinicians should be aware of the emergence of these infections, not only because of the new dietary habits of the population, but also because of the high prevalence of Anisakis larvae in the different fish species usually consumed by the population, collected on the Portuguese coast.

Anisakidosis: report of 25 cases and review of the literature.

Anisakidosis (previously known as anisakiasis) is a disease caused by the accidental ingestion of larval nematodes (Anisakis and sometimes Pseudoterranova) in raw fish. Two groups of patients are studied: 5 clinical cases and 20 serological diagnoses. 55 French cases are already published. Most of them complained of acute symptoms, which occurred within 12 h of eating the seafood meal--epigastric pain, occlusion, diffuse abdominal pain, and appendicitis. Larvae were attached to the gastric mucosa (25 cases), including an inflammatory response (erythema, oedema ulceration). Diagnosis of anisakiasis is made by gastroscopy which allows removal of the worms, and cures the patients. In gastro-intestinal tract X-rays, oedema in the mucosa, pseudo tumour formation, and filling defects (worm) were observed. In chronic infections, cases with intermittent feelings of ill health and abdominal pain, lasting from several weeks to months, were misdiagnosed as another intestinal disease. Positive serological reactions are helpful, and surgery is necessary for resection of the lesion; diagnosis is made histologically by an eosinophilic granuloma, and the presence of a larva with Y shaped lateral cords. Infestation rate is high in fishes: cod (88%), rock fish (86%), herring (88%), salmon, mackerel. Public health education should discourage the eating of raw fish. Thorough cooking to 70 degrees C or adequate freezing to -20 degrees C for 72 h are the best preventive measures. Such legislation is only in force in the Netherlands, where cases have decreased dramatically.

Anisakis simplex induces not only anisakiasis: report on 28 cases of allergy caused by this nematode.

Anisakis simplex R., parasitizes several sea-fish and cephalopods. This parasite induces not only anisakiasis but also anaphylactic reactions, as has recently been confirmed. We present 28 patients with immediate hypersensitivity to Anisakis simplex, developed after parasitized fish ingestion. Each case was diagnosed by suggestive anamnesis, skin prick tests with an A. simplex extract, specific IgE detection in serum (CAP System), and histamine release test. The clinical manifestations were urticaria/angioedema in all 28 patients and respiratory arrest in one. Paradoxically, the patients reported tolerance to ingestion of the same kind of fish between and after the allergic episodes. Type 1 hypersensitivity to A. simplex was demonstrated by a positive skin-prick test reaction, specific IgE detection in all cases and positive histamine release in a representative case. Sensitization to the proteins of the fish themselves was ruled out by negative reactions of the same tests in all cases. It was also proven that the allergen/s involved may be resistant to cooking and deep freezing. Hence, anaphylactic reactions may result either from infection or, more frequently, from mere exposure to the allergen. The results confirm that A. simplex can develop IgE-mediated anaphylactic reactions, the clinical severity of which was pronounced in the patients examined. Physicians must take into account that the consumption of parasitized fish may cause severe reactions, even if fish of the same kind is subsequently tolerated. A higher frequency of reactions of this type can be expected in countries where fish is a staple diet.

Splenic anisakiasis resulting from a gastric perforation: an unusual occurrence.

We report a case of gastric perforation by Anisakis sp. with consequent localization of the larva in the spleen. An 86-year-old white woman was admitted to our surgical department with a diagnosis of acute abdomen. She had a history of abdominal pain, and her laboratory data showed leukocytosis. In the plain abdominal radiograph, pneumoperitoneum was evident; a computed tomography (CT) scan of the abdomen confirmed the presence of extraluminal air. Anamnesis disclosed the ingestion of raw fish during the week preceding her illness. The patient was underwent emergency laparotomy. A small gastric perforation and a nodular area at the superior pole of the spleen were found. Surgical treatment was performed successfully and consisted of excision of the gastric lesion and splenectomy. The histological diagnosis revealed the presence of gastritis with an ulcer, and in the splenic tissue, some necrotic foci containing cross-sectioned degenerated worms compatible with Anisakis larva.

[The problem of anisakiasis]. / Problema anizakidoza.

Presents a brief analysis of the basic findings concerning the pathogenesis, clinical picture, diagnosis, prevention, and epidemiology of anisakiasis in the world. Suggests an opinion on a high prevalence of anisakiasis in some regions of Russia, where it is erroneously diagnosed as some other gastrointestinal diseases. Presents the author's data on the infestation of fish with Anisakis simplex and Pseudoterranova decipiens larvae in the Okhotsk and Bering seas. Emphasizes the necessity of developing recommendations on anisakiasis for practical health service.

Invaginación colocólica por anisakiasis, una causa infrecuente de obstrucción intestinal / Colonic intussusception caused by anisakiasis. A rare cause of obstruction

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Risk factors for human Anisakis infection and association between the geographic origins of Scomber japonicus and anisakid nematodes.

It has been reported that nearly all cases of anisakiasis in Japan are caused by Anisakis simplex sensu stricto. To elucidate this further, we investigated the presence of Anisakis type I larvae and Pseudoterranova decipiens in 218 Scomber japonicus fish collected from the seas of Japan. Anisakis type I larvae were detected in 74.3% (162/218) of the fish, and 99.8% of the Anisakis type I larvae comprised A. simplex sensu stricto and Anisakis pegreffii. Molecular identification techniques were used for 7.5% (360/4806) of the Anisakis type I larvae. The larvae found in the fish of the Pacific stock (the Pacific coast of Japan) and the Tsushima Warm Current stock (the East China Sea and the Sea of Japan) were primarily A. simplex sensu stricto and A. pegreffii, respectively. In addition, for the first time in Japan, Anisakis simplex C and Anisakis ziphidarum were detected in the fish of the Pacific stock. The average number of A. pegreffii and A. simplex sensu stricto larvae per fish was 47 and 6, respectively. However, the average number (0.61 larvae) of A. simplex sensu stricto in the muscle per fish was 12 times the average number (0.05 larvae) of A. pegreffii. When fish on the purchased day were compared with those held at 4 degrees C and 20 degrees C for 20h, the penetration rates (ratio of the number of larvae detected in the muscle to the total number of larvae detected) of A. pegreffii and A. simplex sensu stricto were as high as 1.8% and 5.8%, respectively. In conclusion, we suggest that anisakiasis in Japan is mainly caused by A. simplex sensu stricto because it penetrates the muscle of the fish at a higher rate than A. pegreffii.