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Occupational generalised urticaria and allergic airborne asthma due to anisakis simplex.

Anisakis simplex (AS), a fish and cephalopodes parasite, may cause allergic reactions in humans on eating and/or handling contaminated fish. We present a case of occupational hypersensitivity to AS in a woman employed in a frozen-fish factory. She showed both generalised urticarial rash and asthmatic symptoms after work place exposure. All these symptoms immediately disappeared after work place exposure was ceased. The presence of a positive skin prick test and high specific IgE values confirmed a hypersensitivity to anisakis. This is the first case reported of both occupational generalised urticaria and allergic airborne asthma due to AS in the same patient. We suggest that AS could be an important cause of occupational asthma and/or urticaria in the fish industry.

Air Pollutants, Occupational↗

Freezing protects against allergy to Anisakis simplex.

BACKGROUND: Allergic reactions to Anisakis simplex have been described following ingestion of fish and were thought to be possible even if seafood is frozen or well-cooked. OBJECTIVE: This study aimed to confirm, by challenge test, that dead A. simplex larvae are not able to trigger allergic reactions in patients with proven hypersensitivity to this parasite. METHODS: Simple-blind, placebo-controlled, oral challenge tests with frozen A. simplex larvae were performed in 12 patients who had suffered severe anaphylactic reactions after ingestion of seafood and diagnosed of A. simplex hypersensitivity by skin prick test and specific IgE. If no reaction appeared, they were told to eat frozen seafood. 63 patients who had suffered urticaria or urticaria/angioedema by demonstratred hypersensitivity to A. simplex were also advised to eat frozen seafood. All of them were reevaluated 6 months later. RESULTS: All patients tolerated the dead larvae challenge test. After eating previously frozen seafood at least two times per week, all patients, including those who had suffered anaphylactic reactions and those who had only presented cutaneous manifestations, remained asymptomatic. CONCLUSIONS: Anisakis simplex-allergic patients tolerate ingestion of dead larvae. It is probable that these patients can eat frozen fish and that a seafood-free diet is not necessary.

Adult↗

[Analysis of factors influencing on Anisakis specific IgE antibodies].

Anisakis-specific IgE (AsIgE) was examined in 84 cases of acute urticaria (AU) and 100 cases of atopic dermatitis (AD). Gender, age, disease category (AU or AD) and serum IgE value of the patients were compared with AsIgE. Univariate analysis showed that there was significant correlation between age and AsIgE, and between serum IgE and AsIgE. Serum IgE was significantly higher in AD than in AU. Age was significantly higher in AU than in AD. AsIgE was positive value in 26 cases (31%) of AU and 25 cases (25%) of AD. There were no significant differences between them. In these cases, however, AsIgE was significantly higher in AU than in AD. Percentage of patients with high AsIgE value (> class 2) was significantly higher in AU (20 cases, 24%) than in AD (3 cases, 3%). Multivariate analysis using discriminant analysis and logistic regression analysis showed that the most influential factor on AsIgE was disease category (relative risk = 16.87), and the second was age (relative risk = 1.05). Serum IgE and gender were considered not to influence on AsIgE. It is possible that Anisakis or related antigens have something to do with urticaria in AU patients with high AsIgE value.

Acute Disease↗

Anisakis simplex allergy after eating chicken meat.

BACKGROUND: Allergic reactions to food can be produced by contaminants that induce sensitization. Among these, Anisakis simplex can cause seafood infestation, and allergic symptoms (urticaria-angioedema, anaphylaxis, and asthma) can follow the eating or handling of affected fish. Although seafood is the principal source of human infections by this parasite, we have found allergic symptoms in 8 patients previously diagnosed as having A simplex sensitization after they ate chicken meat. Chicken feed usually has a high proportion of fishmeal, which might possibly be contaminated by this nematode. OBJECTIVE: The aim of our study was to determine whether parasite proteins present in chicken meat could be responsible for the symptoms reported by these subjects. METHODS: We carried out in vivo tests (prick, bronchial challenge, and double-blind placebo-controlled challenge with meat chicken) in these 8 patients. We performed immunoblotting using the sera from the 8 patients and controls in order to detect A simplex sensitization. We also investigated the presence of A simplex proteins in sera from chickens fed with fishmeal and in other sera from chickens fed only with cereals. We excluded sensitization to other chicken nematodes by serologic methods. RESULTS: All 8 patients presented positive prick and challenges to A simplex. When we used serum from chickens fed with fishmeal as the antigen in blotting, patients 3, 4, 5, 6, 7, and 8 recognized a band of 16 kd, also obtained when using pools of fish-shellfish and A simplex larva. No detection was observed with sera from chickens fed with only cereals. CONCLUSION: We provide evidence, based on in vivo and in vitro tests, that subjects highly sensitized to A simplex can detect the presence of Anisakis species allergens in chicken meat.

Adult↗

Stickleback as a potential paratenic host in the Anisakis simplex life cycle in the Baltic Sea: results of experimental infection.

Anisakis simplex stage 3 larvae found in herring were used to experimentally infect 155 sticklebacks and 6 nine-spined sticklebacks, caught in the Gdynia marina (Gulf of Gdańsk). The larvae were observed in the fish body cavity as early as 24 h after infection, only the sticklebacks being affected. Altogether, 50 live and 2 dead larvae were found in the stickleback body cavity. The infection prevalence and mean intensity were 13.6% and 2.4 larvae, respectively, from 1 to 8 larvae per fish being recorded. It is concluded that the stickleback may serve as a paratenic host for Anisakis simplex.

Animals↗

Characterization of the serine protease and serine protease inhibitor from the tissue-penetrating nematode Anisakis simplex.

A serine protease and a serine protease inhibitor were purified from infective larvae of the parasitic nematode Anisakis simplex. The serine protease was found to be trypsin-like and preferentially cleaved substrates with the basic amino acid arginine at the P1 position (Z-Gly-Pro-Arg-AMC (where Z is benzyloxycarbonyl), Km = 0.019 mM, and Z-Phe-Pro-Arg-AMC, Km = 0.013 mM) at rates similar to those determined for trypsin (0.002 mM and 0.006 mM, respectively). However, the presence of a bulky hydrophobic residue at the P2 position (Z-Phe-Arg-AMC, Km = 13.3 mM, and Z-Ile-Leu-Val-Arg-AMC, Km = 24.7 mM) greatly decreased the rate of substrate hydrolysis. Internal amino acid sequence information was obtained from three endo Lys-C digestion fragments of the purified enzyme. These sequences were > 89% (33:37) identical with that of porcine trypsin. A second serine protease 85% (11:13) identical with that of a secreted tissue-destructive serine protease from the pathogenic bacterium Dichelobacter nodosus was also identified. The serine protease inhibitor was found to inhibit trypsin, elastase, and the Anisakis serine protease stoichiometrically, but did not inhibit chymotrypsin. The amino acid sequence of the amino terminus as well as two internal endo Lys-C fragments were determined. Approximately 96% (47:49) of the residues were identical with soybean trypsin inhibitor, indicating that this inhibitor belongs to the Kunitz-type family of inhibitors.

Amino Acid Sequence↗

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.

Adolescent↗

[Allergy to Anisakis simplex. Report of 2 cases and review of the literature].

Anisakis simplex is a nematode which parasitizes mainly fish and cephalopods and accidentally human beings after ingestion. Apart from the different digestive syndromes, this organism has recently been involved as cause of IgE-mediated allergic reactions of variable degree (urticaria to severe angioedema). In this report three patients with the diagnosis of allergy to Anisakis simplex are reported and a review of the literature is made.

Adult↗

Lethal efficacy of extract from Zingiber officinale (traditional Chinese medicine) or [6]-shogaol and [6]-gingerol in Anisakis larvae in vitro.

The authors previously reported that an extract from Zingiber officinale, traditionally eaten along with raw fish and used in traditional Chinese medicine, effectively destroyed Anisakis larvae in vitro. In this study, we analyzed the effective components of ginger rhizomes. Methanol extracts were fractionated after first being treated with HCl at pH 3, then with NaHCO3 at pH 10, and, finally, with NaOH at pH 13 (fraction 1). In general, this fraction is rich in neutral substances. [6]-Shogaol and [6]-gingerol, known neutral components of ginger rhizomes, were detected using gas chromatography and were found to be the most prevalent components in the fraction, occurring in quantities that resulted in a dose-dependent killing efficacy. Authentic [6]-shogaol and [6]-gingerol could kill Anisakis larvae at a minimal effective dose of 62.5 and 250 micrograms/ml, respectively. However, the concentration of [6]-gingerol in fraction 1 was greater than 20 times that of [6]-shogaol, making the former the most active component in the fraction. Furthermore, synergistic effects between [6]-gingerol and a small amount of [6]-shogaol were observed. Pyrantel pamoate, an available antinematodal drug, had no lethal effect, even at a concentration of 1 mg/ml. In saline solution containing [6]-shogaol (62.5 micrograms/ml), greater than 90% of larvae lost spontaneous movement within 4 h and were destroyed completely within 16 h. Microscopical examinations showed destruction of the digestive tract and disturbances of culticulae.

Animals↗

Immunoblot analysis of serum IgG, IgA and IgE responses against larval excretory-secretory antigens of Anisakis simplex in patients with gastric anisakiasis.

To increase our understanding of the immune response to Anisakis infection, antigen specific IgG, IgA and IgE responses were identified using an immunoblot technique after polyacrylamide gel electrophoresis of excretory-secretory products from the larval stage of Anisakis simplex. Nine sera were drawn from proven cases of gastric anisakiasis within 3 days after symptoms had developed. The molecular weight of the major antigenic bands were distributed between 50 kDa and 120 kDa of the antigens. In nine cases of gastric anisakiasis, three of them were positive for IgG response, five for IgE, and six for IgA, respectively. None of control sera recognized the antigenic bands in IgA and IgE responses. In contrast, two controls had IgG antibodies against 1-2 proteins in the 65-95 kDa region. The antigenicity of the excretory-secretory products was lost following treatment by 0.2% trypsin, but not by 0.2 M periodic acid. Based on the results of reactivity to lectins, antigenic bands of the ES products possessed mucin type glycoconjugate residues in their protein portion. This indicates that the humoral responses of IgA and IgE antibodies to the larval ES antigens are a more reliable index of infection than that of the IgG response.

Adult↗

[A case report on human infection with Anisakis sp. in Korea]

Authors experienced a case of human infection with Anisakis sp. in left tonsil. The case was 27 year Korean female who visited Department of Otolaryngology, Severance Hospital with the chief complaints of swollowing difficulty, and the sensation of a foreign body in the throat, on November 20th 1968. Physical examination of the oral cavity revealed that both palatin tonsil were chronically enlarged. A larval worm was found in crypt of the upper portion of the left palatin tonsil. Chief complaints of the patient was improved after removal of it by forceps. The greyish white living worm was fixed in lactophenol solution for morphological study. The worm measured 36 mm in length by 0.6 mm in width, 2.4 mm in esophageal length, 1.4 mm in ventricule, 0.1 mm in tail length, and 0.025 mm in caudal spine length, respectively and histological study was carried out from the serial section of the larva which was stained with hematoxylin eosin. Authors identified that the larva was Anisakis species.

Journal Article↗

Survey of Anisakis larvae in marine fish of Taiwan.

A survey on the prevalence and intensity of larval anisakid infection in some species of marine fishes sold in the markets at Tan-shui in northern Taiwan was carried out from May to August 1980. The general infection rate was 37.7% with an average of 14.2 larvae per fish. A total of 13 species of examined fishes were found with Anisakis larvae, these were Argyrosomus argentatus, Caranx djeddaba, Diploprion bifasciatum, Evynnis cardinalis, Lethrinus haematopterus, Megalops cyprinoides, Nemipterus virgatus, Paraplagusia formosana, Plectorhinchus pictus, Rastrelliger chrysozonus, Saurida tumbil, Scolopsis vosmeri, and Trichiurus haumela. The highest intensity of Anisakis larvae was obtained in E. cardinalis with an average of 80.3 larvae per fish; the next was in N. virgatus (76.2 larvae/fish). The parasite could not be found in the other 20 species of fishes examined.

Animals↗

Occurrence and survival of the larval nematode Anisakis sp. in the flesh of fresh, frozen, brined, and smoked pacific herring, Clupea harengus pallasi.

Two hundred and twenty Pacific herring (Clupea harengus pallasi) from Yaquina Bay, Oregon, were divided into five groups to test the effects of various methods of handling and processing (fresh, frozen, brined, cold smoked, and cold smoked-gibbed) on the occurrence and survival of Anisakis larvae in the flesh. Viscera and flesh were digested separately using a pepsin-HC1 technique, and counts were taken of Anisakis larvae. Parasite loads were statistically largest in the flesh of frozen, brined, and smoked fish. Smoked fish harbored the greatest proportions of larvae in the flesh. Human consumption of brined or cold smoked Pacific herring represents a potential public health hazard.

Animals↗

Early gastric cancer superimposed on infestation of an Anisakis-like larva: a case report.

This report describes a 73-year-old male with early gastric cancer (type IIc) superimposed on infestation of a parasitic larva. Eosinophilic granulomas surrounding a dead worm were seen in the submucosa just beneath the intramucosal tubular adenocarcinomatous lesion measuring 1 X 1cm at the anterior wall in the acid-secreting area. The cancer cells showed lowered and altered mucin production in comparison with the surrounding non-cancerous fundic mucosa. The parasite was identified as an Anisakis-like larva by the presence of typical lateral chords. An immunohistochemical examination revealed that the cancer cells were more strongly positive for IgA and secretory component than the surrounding non-cancerous mucosa, and that IgG, IgA and IgM were detected in plasma cells around the granulomas but IgE and IgD were not. The possibility that the Anisakis-like larva preferentially infested the cancerous mucosa because of the change in mucin, local defect in acid secretion and/or other structural alterations in the area is discussed.

Adenocarcinoma↗

[1st findings of Anisakis larvae in pike perches, Stizostedion lucioperca, in coastal waters of the Baltic Sea].

For the first time Anisakis larvae were found in pike perches, Stizostedion lucioperca (L), caught in GDR coastal waters. Herrings parasitized by Anisakis are regarded as the source of infestation, for herrings represent an important part of the food of the pike perch. The occurrence of anisakiasis by eating pike perch is very unlikely considering eating habits in the GDR. The occurrence of larvae as "natural indicators" can be applied in fish biological research.

Animals↗

A PCR-RFLP assay for identification of Anisakis simplex from different geographical regions.

Anisakis simplex, a nematode from the family Anisakidae, is a parasite of fish and mammals. It is a casual agent of a human disease called anisakiosis. We found that the assay based on PCR amplification of the ITS-1-5.8 S-ITS-2 fragment of rDNA and subsequent restriction fragment length polymorphism, previously described on the basis of A. simplex isolated solely from one geographical region, can be used as a general test for identification of this worm species. The restriction patterns analysed for four restriction enzymes were found to be identical in the case of all A. simplex individuals isolated from as different geographical regions as Baltic Sea, Norwegian Sea, Bering Sea and Sea of Okchotsk. Moreover, our results support the previously proposed hypothesis, based on the studies of isoenzymes, that there is a remarkable genetic homogeneity within A. simplex from different geographical regions.

Animals↗

A recombinant enolase from Anisakis simplex is differentially recognized in natural human and mouse experimental infections.

A 1,963-bp cDNA was isolated from an Anisakis simplex cDNA library by immunoscreening with a hyperimmune rabbit serum raised against a crude extract of A. simplex L3 larvae. The open reading frame encodes a putative protein of 436 amino acid residues, which exhibits high similarity (70-80%) to enolase molecules from various other organisms, including helminth parasites. After subcloning and expression of the A. simplex cDNA in PGEX-4T-3, the resulting glutathione S-transferase fusion protein, purified by glutathione-Sepharose-4B chromatography, showed functional enolase activity. The immunogenicity of the recombinant A. simplex enolase was analyzed by immunoblotting using sera obtained from (a) mice immunized with crude extracts (CE) of A. simplex, or other nematode species, (b) mice immunized with excretory-secretory (ES) antigens from A. simplex, or (c) mice infected with L3 larvae by the intraperitoneal route. In addition, we used ELISA, to investigate the presence of IgG1 and IgE antibodies against this molecule in sera from patients infected with A. simplex. Mouse sera obtained after infection with L3 or raised against CE antigens, but not sera raised against ES antigens, showed strong reactivity with the recombinant A. simplex enolase. We also obtained good reactivity in Western blotting with sera from mice immunized with CE antigens from Ascaris suum and Toxocara canis, but not with sera from mice immunized with CE antigens from Trichuris muris, Trichinella spiralis or Hysterothylacium aduncum. In contrast to the experimental infections/immunizations in mice, we were unable to detect anti-enolase IgE antibodies in sera from human patients infected with A.simplex (15 sera), and the levels of anti-enolase IgG1 antibodies in these sera were low and apparently nonspecific. These results seem to indicate that, during natural infection in humans, A. simplex larvae do not offer sufficient antigenic stimulus to induce anti-enolase antibodies.

Animals↗

Ultrastructural localization of Ani s 1, a major allergen from the fish parasite Anisakis simplex.

No data about the nature or function of the first major Anisakis simplex allergen, named Ani s 1, is available. The aim of this study was to investigate the ultrastructural localization of this protein, to obtain further information about it. Ani s 1 was detected in secretory granules of the excretory gland and occasionally lining the main excretory canal. Our results suggest that Ani s 1 could be a secretory product and could have an enzymatic function related to mechanisms of infection.

Allergens↗