Expansion of T regulatory lymphocytes by murine bone marrow dendritic cells previously stimulated with Anisakis simplex larval antigens

BACKGROUND Anisakis simplex antigens present immunomodulatory properties by the induction of tolerogenic dendritic cells (DCs) in mice. OBJECTIVES To study the capacity of DCs stimulated with A. simplex excretory-secretory (ES) or crude extract (CE) to generate Tregs. To investigate in vitro effects of antigens on the metabolic activity of splenocytes induced by LPS or CpG. METHODS Phenotypic and functional characterization of T cells co-cultured with A. simplex-pulsed DCs was performed by flow cytometry. Lymphocyte mitochondrial respiratory activity was estimated by the Alamar Blue® Assay. FINDINGS In C57BL/6J, CD4+CD25-Foxp3+ and CD8+CD25-Foxp3+ populations increased by CE-stimulated-DCs. In BALB/c, CE-stimulated-DCs caused the expansion of CD4+CD25+Foxp3+IL-10+ and CD8+CD25+Foxp3+IL-10+. IFN-γ expression raised in BALB/c CD4+CD25+ and CD4+CD25- for CE and ES, respectively. ES-stimulated-DCs increased CD4+CD25+ Foxp3+ and CD8+CD25- Foxp3+ expression in T cells. The association of ES or CE with LPS produced the increase in splenocyte activity in C57BL/6J. The association of CE with CpG decreased the proliferation caused by CpG in C57BL/6J. MAIN CONCLUSIONS A. simplex increase the frequency of Tregs, which in turn produce IL-10 and IFN-γ. The host genetic base is essential in the development of anti-Anisakis immune responses (Th2, Th1, Treg).

Molecular Identification of Anisakis Larvae Extracted by Gastrointestinal Endoscopy from Health Check-up Patients in Korea

Anisakiasis is a zoonotic disease induced by anisakid nematodes, and endoscopic inspection is used for a diagnosis or remedy for it. Anisakis simplex, Anisakis physeteris, and Pseudoterranova decipiens had been reported to be the major species causing human infections, particularly, in Japan. However, in Korea, recent studies strongly suggested that Anisakis pegreffii is the major species of human infections. To support this suggestion, we collected anisakid larvae (n=20) from 20 human patients who were undergone gastrointestinal endoscopy at a health check-up center in Korea, and molecular identification was performed on the larvae using PCR-RFLP analysis and gene sequencing of rDNA ITS regions and mtDNA cox2. In addition, anisakid larvae (n=53) collected from the sea eel (Astroconger myriaster) were also examined for comparison with those extracted from humans. The results showed that all human samples (100%) were identified as A. pegreffii, whereas 90.7% of the samples from the sea eel were A. pegreffii with the remaining 9.3% being Hysterothylacium aduncum. Our study confirmed that A. pegreffii is the predominant species causing human anisakiasis in Korea, and this seems to be due to the predominance of this larval type in the fish (sea eels) popularly consumed by the Korean people. The possibility of human infection with H. aduncum in Korea is also suggested.

Anisakidae nematodes in the blackfin goosefish, Lophius gastrophysus Miranda-Ribeiro, 1915 purchased in the State of Rio de Janeiro, Brazil / Nematoides Anisakidae no peixe sapo-pescador, Lophius gastrophysus Miranda-Ribeiro, 1915 comercializado no Estado do Rio de Janeiro, Brasil

The blackfin goosefish is included in a prized fish category, representing valuable fishery resource worldwide. The aim of this study was to identify the Anisakidae larvae parasitizing this fish species, considering the hygienic-sanitary and public health importance of these parasites infecting specimens of the blackfin goosefish, Lophius gastrophysus Miranda-Ribeiro, 1915 (Lophiidae), purchased from markets in the municipalities of Cabo Frio, Niterói, Duque de Caxias and Rio de Janeiro; parasitological indices and sites of infection are presented. From March 2000 to December 2005, 87 specimens of Lophius gastrophysus were investigated for helminths. Seventeen fish (19.5%) were parasitized with larvae of anisakid nematodes. Anisakis simplex , Hysterothylacium sp. and Raphidascaris sp. appeared with prevalence of 1.14, 12.6 and 12.6%, intensity and mean intensity of infection of 1, 2.81, 10.5 and mean abundance of 0.01, 0.35 and 1.33 respectively. The sites of infection were stomach, stomach and intestine serosas and abdominal cavity. This is the first record of Anisakis simplex, Hysterothylacium sp. and Raphidascaris sp. for this species of fish.

Nematoides Anisakidae no peixe sapo-pescador, Lophius gastrophysus Miranda-Ribeiro, 1915 comercializados no Estado do Rio de Janeiro, Brasil. O peixe sapo-pescador é muito apreciado e representa um valioso recurso pesqueiro em nível mundial. O objetivo deste estudo foi o de identificar larvas de Anisakidae pela sua importância higiênico-sanitária e na saúde pública, cujos parasitos infectam espécimes do peixe sapo-pescador, Lophius gastrophysus Miranda-Ribeiro, 1915 (Lophiidae), obtidos em mercados dos municípios de Cabo Frio, Niterói, Duque de Caxias e Rio de Janeiro; são apresentados os índices parasitológicos e sítios de infecção dos helmintos encontrados. Foram investigados 87 espécimes de Lophius gastrophysus no período de março de 2000 a dezembro de 2005. Desses, 17 (19,5%) estavam parasitados por larvas de anisaquídeos. Anisakis simplex, Hysterothylacium sp. e Raphidascaris sp. apareceram com prevalências de 1,14, 12,6 e 12,6%, intensidade e intensidade média de infecção de 1, 2,81 e 10,5, abundância média de 0,01, 0,35 e 1,33 respectivamente. Os sítios de infecção foram o estômago, serosas do estômago e intestino e cavidade abdominal. Este é o primeiro registro de Anisakis simplex, Hysterothylacium sp. e Raphidascaris sp. nesta espécie de peixe.

Anisakiose: soropositividade humana e desenvolvimento de um modelo experimental / Anisakiasis: human seropositivity and development of an experimental model

Existem evidências científicas de que várias espécies de peixes do litoral do Estado do Rio de Janeiro estão contaminadas com a larva nematoide Anisakis simplex. A infecção em humano, a sororeatividade e a doença ainda não são bem conhecidas em território brasileiro. Também os modelos de estudo sa anisakiose experimental atuais em camundongos tem utilizado como norma geral a introdução da forma larvária ou de seu extrato antigênico por via intraperitoneal... Foram observados sororeatividade para antígenos somáticos e para os secretores/excretores apresentando valores significativamente mais elevados quando comparados aos controles (p,0,001) quando analisados por ANOVA e pós teste de Tukey. A relação direta entre a reatividade no ELISA e a frequência da ingestão sem relação com sintomas alérgicos sugerem a existência de contato prévio com antígenos do Anisakis simplex, mas não podem atestar uma tendência alérgica no grupo analisado. As taxas de reatividade em puérperas com pequena ingestão reforça esse raciocínio. Em animais, através deste experimento foi possível demonstrar a efetividade da utilização da via gástrica para sensibilização de uma infecção intestinal por larva viva de Anisakis simplex em camundongos.

Preventive and Therapeutic Effects of Anisakis simplex Larval Protein in a Mouse Model of Crohn'S Disease / 고신대학교의과대학학술지

OBJECTIVES: Some helminths have been known to have a treatment effect in inflammatory bowel diseases, including Crohn's disease (CD); however, live parasite therapy can cause unwanted side effects. To develop a safe therapeutic, we investigated the preventive or therapeutic potential of proteins from the third stage larva of A. simplex in a mouse model. We also analyzed the cytokine profile from splenic and mesenteric lymph node lymphocytes to elucidate the underlying immunological mechanism. METHODS: CD was induced in mice with DSS, and the effect of an A. simplex larval protein on CD was assessed. A change in body weight and DAI (disease activity index) were observed in mice. The expression levels of cytokines from mesenteric lymph nodes (MLN) compared to splenic lymphocytes were measured with ELISA. RESULTS: Peritoneal administration of preventive and therapeutic A. simplex larval proteins attenuated DSS-induced CD by a reduction of the DAI and weight loss. A shortening of colon length was more definitely observed in the therapeutic group than in the preventive group. The cytokine expression levels were more obvious in lymphocytes from mesenteric lymph nodes than from splenic lymphocytes. CONCLUSIONS: Taken together, these results suggest that A. simplex proteins can change cytokine profiles and may have a preventive effect in DSS-induced CD mice.

Anisakidosis in Korea; Changes Over the Last Decade / 고신대학교의과대학학술지

Anisakidosis is caused by Anisakis simplex and other anisakids larvae parasitizing marine fish and cephalopods. A lot of case reports about anisakidosis have been published in Korea because of raw fish eating habits. Recently seafood consumption has continued to increase due to health concerns and thus, it increases the risk for infectious diseases including anisakidosis. The aim of this review is to analyze the clinical and epidemiological characteristics of anisakidosis during the last 10 years in Korea, based on the case reports published from 2000 to 2010. The incidence age was changed from 30s and 40s to 50s. The young generation was considered to consume seafood in various ways, including raw fish as well. The most noticeable change was the appearance of Anisakis allergy patients over the last decade. The patients showed abdominal pain, urticaria after eating sea food. It reaffirmed that anisakid infection induces not only gastric and intestinal anisakidosis but also cause allergic reaction. Anisakid should be considered as a possible causative food allergen provoking allergic responses after eating raw fish.

Larval Anisakid Infections in Marine Fish from Three Sea Areas of the Republic of Korea

The present study was performed to determine the infection status of anisakid larvae in marine fish collected from 3 sea areas of the Republic of Korea. Total 86 marine fish (8 species) collected from the East Sea (Goseong-gun, Gangwon-do), 171 fish (10 species) from the South Sea (Sacheon-si, Gyeongsangnam-do), and 92 fish (7 species) from the Yellow Sea (Incheon Metropolitan City) were examined by both naked eyes and artificial digestion method. Among the total of 349 fish examined, 213 (61.0%) were infected with 8 species of anisakid larvae, i.e., Anisakis simplex, 6 types of Contracaecum spp., and Raphidascaris sp., and the mean larval density was 13.8 per infected fish. Anisakid larvae were detected in 45 fish (52.3%) from the East Sea, 131 fish (76.6%) from the South Sea, and 37 fish (40.2%) from the Yellow Sea. The average numbers of larvae detected were 4.0, 16.6, and 15.9, respectively. Anisakis simplex larvae were detected in 149 fish (42.7%), and the mean larval density was 9.0 per infected fish. They were found in 26 fish (30.2%) collected from the East Sea, 96 fish (56.1%) from the South Sea, and 27 fish (29.3%) from the Yellow Sea. The average numbers of larvae detected were 2.9, 10.3, and 10.5, respectively. Conclusively, the present study suggests that the infection rate and density of anisakid larvae are more or less higher in the fish from the South Sea than those from the East Sea or the Yellow Sea.

Expression and Characterization of alpha-Methylacyl CoA Racemase from Anisakis simplex Larvae

Larval excretory-secretory products of Anisakis simplex are known to cause allergic reactions in humans. A cDNA library of A. simplex 3rd-stage larvae (L3) was immunoscreened with polyclonal rabbit serum raised against A. simplex L3 excretory-secretory products to identify an antigen that elicits the immune response. One cDNA clone, designated as alpha-methylacyl CoA racemase (Amacr) contained a 1,412 bp cDNA transcript with a single open reading frame that encoded 418 amino acids. A. simplex Amacr showed a high degree of homology compared to Amacr orthologs from other species. Amacr mRNA was highly and constitutively expressed regardless of temperature (10-40degrees C) and time (24-48 hr). Immunohistochemical analysis revealed that Amacr was expressed mainly in the ventriculus of A. simplex larvae. The Amacr protein produced in large quantities from the ventriculus is probably responsible for many functions in the development and growth of A. simplex larvae.

Anisakis simplex Larvae: Infection Status in Marine Fish and Cephalopods Purchased from the Cooperative Fish Market in Busan, Korea

The infection status of marine fish and cephalopods with Anisakis simplex third stage larva (L3) was studied over a period of 1 year. A total of 2,537 specimens, which consisted of 40 species of fish and 3 species of cephalopods, were purchased from the Cooperative Fish Market in Busan, Korea, from August 2006 to July 2007. They were examined for A. simplex L3 from the whole body cavity, viscera, and muscles. A. simplex L3 were confirmed by light microscopy. The overall infection rate reached 34.3%, and average 17.1 larvae were parasitized per infected fish. Fish that recorded the highest infection rate was Lophiomus setigerus (100%), followed by Liparis tessellates (90%), Pleurogrammus azonus (90%), and Scomber japonicus (88.7%). The intensity of infection was the highest in Gadus macrocephalus (117.7 larvae per fish), followed by S. japonicus (103.9 larvae) and L. setigerus (54.2 larvae). Although abundance of A. simplex L3 was not seasonal in most of the fish species, 10 of the 16 selected species showed the highest abundance in February and April. A positive correlation between the intensity of L3 infection and the fish length was obvious in S. japonicus and G. macrocephalus. It was likely that A. simplex L3 are more frequently infected during the spring season in some species of fish. Our study revealed that eating raw or undercooked fish or cephalopods could still be a source of human infection with A. simplex L3 in Korea.

A 24 kDa Excretory-Secretory Protein of Anisakis simplex Larvae Could Elicit Allergic Airway Inflammation in Mice

We have reported that a 24 kDa protein (22U homologous; As22U) of Anisakis simplex larvae could elicit several Th2-related chemokine gene expressions in the intestinal epithelial cell line which means that As22U may play a role as an allergen. In order to determine the contribution of As22U to allergic reactions, we treated mice with 6 times intra-nasal application of recombinant As22U (rAs22U). In the group challenged with rAs22U and ovalbumin (OVA), the number of eosinophils in the bronchial alveolar lavage fluid (BALF) was significantly increased, as compared to the group receiving only OVA. In addition, mice treated with rAs22U and OVA showed significantly increased airway hyperresponsiveness. Thus, severe inflammation around the airway and immune cell recruitment was observed in mice treated with rAs22U plus OVA. The levels of IL-4, IL-5, and IL-13 cytokines in the BALF increased significantly after treatment with rAs22U and OVA. Similarly, the levels of anti-OVA specific IgE and IgG1 increased in mice treated with rAs22U and OVA, compared to those treated only with OVA. The Gro-alpha (CXCL1) gene expression in mouse lung epithelial cells increased instantly after treatment with rAs22U, and allergy-specific chemokines eotaxin (CCL11) and thymus-and-activation-regulated-chemokine (CCL17) gene expressions significantly increased at 6 hr after treatment. In conclusion, rAs22U may induce airway allergic inflammation, as the result of enhanced Th2 and Th17 responses.

Seroprevalence of Antibodies against Anisakis simplex Larvae among Health-Examined Residents in Three Hospitals of Southern Parts of Korea

The present study was performed to estimate the seroprevalence of larval Anisakis simplex infection among the residents health-examined in 3 hospitals in southern parts of Korea. A total of 498 serum samples (1 serum per person) were collected in 3 hospitals in Busan Metropolitan city, Masan city, and Geoje city in Gyeongsangnam-do (Province) and were examined by IgE-ELISA and IgE-western blotting with larval A. simplex crude extract and excretory-secretory products (ESP). The prevalence of antibody positivity was 5.0% and 6.6% with ELISA against crude extracts and ESP, respectively. It was also revealed that infection occurred throughout all age groups and higher in females than in males. A specific protein band of 130 kDa was detected from 10 patients with western blot analysis against crude extract and ESP among those who showed positive results by ELISA. Our study showed for the first time the seroprevalence of anisakiasis in Korea. The allergen of 130 kDa can be a candidate for serologic diagnosis of anisakiasis.

Anisakiosis / Anisakiosis

Anisakis simplex es un nematodo de distribución mundial que infecta a los consumidores de pescado crudo o poco cocido. El ciclo vital del parásito puede incluir uno o más hospedadores intermediarios y como hospedadores definitivos mamíferos marinos y grandes peces en los cuales la larva se desarrolla hasta alcanzar el estadio adulto. El hombre es un huésped accidental. Las manifestaciones clínicas de la anisakiosis dependen del lugar del tracto digestivo en que la larva se deposite. Los síntomas se desarrollan como resultado de procesos inflamatorios que se producen en la mucosa de la pared gástrica cuando penetra en ella la larva. Para prevenir esta patología no se debe consumir pescado crudo o que no haya sido congelado previamente durante 48 h, ni pescado fresco que no haya sido cocinado al menos durante 20 min. a más de 60 °C. La incidencia de la anisakiosis está en aumento en los últimos tiempos, por lo que resulta importante conocer sus manifestaciones más características, tanto clínicas como epidemiológicas y de laboratorio, especialmente porque en Venezuela hay elevado consumo de pescado, aunque no se tenga casos. Las manifestaciones más frecuentes son urticaria/angioedema recidivante y dolores abdominales ¿Cuántos de estos síntomas quedan sin diagnóstico en las consultas médicas diarias?.

Anisakis simplex is a widely distributed nematode infecting consumers of raw or undercooked fish. Life cycle of the parasite might include one or more intermediate hosts and sea mammals and big fishes as definitive hosts in which larvae develop to its adult stage. Man is an accidental host. Clinical manifestations of anisakiasis depend on the site of dwelling of larvae in the digestive tract. Symptoms develop as the result of an inflamatory process in the gastric mucosae wall due to the penetration of larvae. Prevention of this pathology relies in avoiding ingestion of raw fish or fish that hasn't been previously frozen at least 48 hours or cooked during at least 20 minures above 60º C. Incidence of anisakiasis is increasing, thus it is important to know and discuss its clinical, epidemiological and laboratory patterns, specially in Venezuela, where we register a high ingestion of fish without case register. Most common clinical features are: urticaria, recidivant angio-oedema and abdominal pain.

The Clinical Characteristics of Anisakis Allergy in Korea

Anisakidae larvae can cause anisakiasis when ingested by humans. Although several groups have reported a gastrointestinal Anisakis allergy among people in Spain and Japan, our report is the first to summarize the clinical features of 10 Anisakis allergy cases in Korea. We enrolled 10 Korean patients (6 men and 4 women) who complained of aggravated allergic symptoms after ingesting raw fish or seafood. Sensitization to Anisakis was confirmed by detecting serum specific IgE to Anisakis simplex. The most common manifestation of anisakiasis was urticaria (100%), followed by abdominal pain (30%) and anaphylaxis (30%). All patients presenting with these symptoms also exhibited high serum specific IgE (0.45 to 100 kU/L) to A. simplex. Nine patients (90%) exhibited atopy and increased total serum IgE levels. The fish species suspected of carrying the Anisakis parasite were flatfish (40%), congers (40%), squid (30%), whelk (10%), and tuna (10%). Anisakis simplex should be considered as a possible causative food allergen in adult patients presenting with urticaria, angioedema, and anaphylaxis following the consumption of raw fish or seafood.

Reactividad alérgica a Anisakis simplex y su asociación con asma bronquial en niños escolares del estado Nueva Esparta, Venezuela / Anisakis simplex allergic reactivity and its association with bronchial asthma in scholar children from Nueva Esparta state, Venezuela

La reactividad alérgica frente a Anisakis simplex se ha convertido en un problema de salud pública en muchos países del mundo. En Venezuela, se ha reportado la presencia de mamíferos marinos infectados y se ha evidenciado la presencia del parásito en pescados de consumo masivo procedentes de las zonas costeras de oriente y occidente del país. No obstante, la importancia de este helminto en el desarrollo y severidad de patologías alérgicas no ha sido evaluada en el país. El objetivo de este estudio fue evaluar la reactividad alérgica al An. simplex en un grupo de 144 niños del estado de Nueva Esparta, así como su posible relación con la severidad de asma bronquial y rinitis alérgica. Todos los niños fueron evaluados utilizando un cuestionario socioeconómico validado y modificado de Graffar y un cuestionario validado según los criterios de ARIA (International Consensus Report) para rinitis alérgica, GINA (WHO/NIH Global Initiative for Asthma) para asma bronquial y Hannifin para dermatitis atópica, respectivamente. Se realizaron encuestas para evaluar las principales fuentes de alergenos a los cuales se expone la población de estudio. Se realizó la evaluación física de los niños y se practicaron pruebas cutáneas, así como pruebas de funcionalismo pulmonar. Encontramos un alto porcentaje (45%) de niños con pruebas de piel positiva al An. simplex. Se observó una mayor prevalencia y severidad, de asma y rinitis en aquellos niños con pruebas de piel positivas a An. simplex. Dado que ha sido previamente reportada la reactividad cruzada entre Ascaris lumbricoides y An. simplex, es importante considerar que la alta reactividad encontrada frente al An. simplex pueda deberse a la co-infección por Ascaris. De igual forma, la sensibilización frente a An. simplex, ya sea especifica o por reactividad cruzada existe, lo que sugiere que las autoridades locales deben considerar ciertas medidas preventivas, para evitar el consumo de pescados crudos o posiblemente parasitados con este helminto, que podrían exacerbar la patología respiratoria.

Allergic reactivity to Anisakis simplex has become in a health problem in many parts of the World. In Venezuelan coasts, it has been found infecting marine mammals and also different parasites fishes commonly consumed by human. However, the association of this helminth with the development and severity of allergic reactivity has not been previously evaluated in Venezuela. The aim of this study was to evaluate the allergic reactivity to An. simplex in a group 144 children from Nueva Esparta, state, and its possible relation with bronquial asthma and rhinitis allergic severity. All children were evaluated using a validated socioeconomic questionnaire modified by Graffar, and a validate questionnaire according to ARIA (International Consensus Report) for allergic rhinitis, GINA (WHO/NIH Global Initiative for Asthma) for bronquial asthma and Hannifin for atopic dermatitis. Surveys were performed in order to evaluate the main sources of allergens to which the study population exposes itself. Children physical evaluation and skin prick test were performed, as well as pulmonary function test. We found a high percentage (45%) of children with positive skin prick test to An. simplex. Moreover, a greatest prevalence and severity of asthma and rhinitis, was observed in those children with positive skin prick test to An. simplex. Since it has been reported the presence of the crossed reactivity between Ascaris lumbricoides and An. simplex, it’s important to consider that the high reactivity found to An. simplex might be due to the Co-infection with A. lumbricoides. Nevertheless, the sensibilization to An. simplex, due by specific or by crossed reactivity exists, and it should be considered by local health authorities in order to prevent the exacerbation of the respiratory pathologies mediated by the consumption of fresh fishes potentially infected.

Pseudoterranovosis y sushi / Pseudoterranovosis and sushi

Se comunica un caso de ingestión accidental de Pseudoterranova decipiens. A raíz de esta paciente se revisan los principales conceptos relativos a esta naciente patología en Chile. El consumo de platos preparados con pescados crudos como el sushi, sashimi, cebiche y mariscal es cada vez más popular. A través de la ingesta de hospederos intermediarios usados en la preparación, se puede adquirir la larva de tercer estadio de Anisakis simplex o P. decipiens. Estos parásitos pueden provocar gastritis aguda que se manifiesta inmediatamente después de consumido el pescado crudo. En ocasiones puede haber compromiso intestinal e invasión del tracto gastrointestinal con manifestaciones en otros órganos. Se asocia a manifestaciones alérgicas como urticaria, angioedema y shock anafiláctico. La larva se elimina en forma espontánea en la mayoría de los casos, pero en otros debe removerse por vía endoscópica. En Chile, es más frecuente la presencia de P. decipiens, que por lo general no migra a la mucosa gástrica, no invade y produce pocos síntomas. Las medidas de prevención son no comer pescados crudos, cocinarlos a temperaturas elevadas o congelación previa a la preparación de los alimentos, para inactivar las larvas.

A case of accidental ingestión of Pseudoterranova decipiens is reported, on account of this patient we review main concepts related to this emerging pathology in Chile. Consumption of raw fish in different preparations such as sushi or sashimi, cebiche and seafood is becoming more and more popular. Through intermediary hosts used in the preparation, one can acquire the third stage larval of Anisakis simplex or P. decipiens. These parasites frequently cause gastritis, acutely after the consumption of raw fish, or more rarely intestinal involvement. It may also cause allergic symptoms such as urticaria, angioedema and anaphylactic shock. The larvae is eliminated spontaneously in most cases; others need to be removed by upper gastrointestinal endoscopio examination. In Chile, the most common species is P. decipiens, which are generally no invasive, rarely migrate beyond the stomach and cause mild or no symptoms. Prevention strategies include avoid eating raw fish or undercooked seafood containing the parasite, to cook food with high temperatures or to freeze raw fish prior to preparation of meals, which inactivates the larvae.

Immune reactions and allergy in experimental anisakiasis

The third-stage larvae (L3) of the parasitic nematode, Anisakis simplex, have been implicated in the induction of hyperimmune allergic reactions in orally infected humans. In this work, we have conducted a review of an investigation into immune reactions occurring in animals experimentally infected with A. simplex L3. The patterns of serum antibody productions in the experimental animals against excretory-secretory products (ESP) of A. simplex L3 contributed to our current knowledge regarding specific humoral immune reactions in humans. In our review, we were able to determine that L3 infection of experimental animals may constitute a good model system for further exploration of immune mechanisms and allergy in anisakiasis of humans.

A case of acute gastric anisakiasis provoking severe clinical problems by multiple infection

Acute gastric anisakiasis with multiple anisakid larvae infection is reported. A 68-year-old woman residing in Busan, Korea, had epigastric pain with severe vomiting about 5 hours after eating raw anchovies. Four nematode larvae penetrating the gastric mucosae in the great curvature of the middle body and fundus were found and removed during gasteroendoscopic examination. Another one thread-like moving larva was found in the great curvature of upper body on the following day. On the basis of their morphology, the worms were identified as the 3rd stage larvae of Anisakis simplex. This case is acute gastric anisakiasis provoking severe clinical problems by the multiple infection and the greatest number of anisakid larvae found in a patient in Korea.

Survival of the Third Stage Larvae of Anisakis simplex in Various Conditions and Experimental Infection in Rats / 中国寄生虫学与寄生虫病杂志

Objective To observe the characteristics of biology and pathogenicity of the third stage Anisakis simplex larvae ( L3). Methods The survival time of the L3 in various conditions was observed and the pathological change after experimental infection in rats was examined. Results The results showed that the L3 frozen at -20℃ for 10 - 12 h can be killed. In the temperature range of 4 - 10 ℃ , the L3 can survive for over 8 months. The L3 was very active at 37℃ , and was killed at the high temperature over 40℃ in a very short time. The ingredients for sashimi cannot kill the L3. The experiment of rats infected by the L, showed that about 15% -25% of the L3 penetrated into the gastrointestinal wall or migrated into the peritoneal cavity in 2 days. After 3d the L4 was not infectious, and died automatically in 7 - 10 days and could not develop into adults. The animals can be easily infected when the stomach was empty. The pathological study showed that the primary infection was a kind of reaction to foreign body, while that of the re - infection was allergic. Conclusion The L3 has a strong resistance to low temperature and to ingredients , it can be killed by freezing at -20℃ in 24 hours. The L3 can not mature in the body of terrestrial mammals but causes pathological change in the stomach and allergy.[