The Inhibitory Effect of Wood Creosote on the Movement of Anisakis Larvae: An Implication for the Treatment of Acute Anisakiasis.

INTRODUCTION: Anisakiasis is a common disease in countries such as Japan, where raw or undercooked marine fish are frequently consumed. The disease is caused by accidental ingestion of a live larva of Anisakis in raw or undercooked marine fish. In typical cases, it causes abrupt gastrointestinal symptoms, such as epigastric pain, nausea, and vomiting. According to a published report, the disease was alleviated by oral ingestion of an over-the-counter drug containing wood creosote. METHODS: We performed an in vitro experiment to elucidate whether wood creosote can inhibit the motor activity of Anisakis larvae, using infrared locomotion tracking and agarose gel penetration techniques. RESULTS: Our results clearly demonstrate that wood creosote inhibits the motor activity of Anisakis larvae. The concentration of wood creosote used in our experiment is similar to that found in stomach juice when a usual oral dose is taken of the medicine containing wood creosote. DISCUSSION/CONCLUSION: Our results suggest the potential usefulness of the medicine containing wood creosote in the treatment of acute Anisakis infection of the gastrointestinal tract.

The oil of garlic, Allium sativum L. (Amaryllidaceae), as a potential protectant against Anisakis spp. Type II (L3) (Nematoda) infection in Wistar rats.

The consumption of inadequately thermally treated fish is a public health risk due to the possible propagation of Anisakis larvae. The present study demonstrated the physiological and histopathological changes that accompanied an oral inoculation of crude extracts from fresh and thermally treated Anisakis Type II (L3) in rats. Worms were isolated from a marine fish and examined and identified using light and scanning electron microscopy. The study was performed in 6 rat groups: control (I), garlic oil (GO) inoculated (II), fresh L3 inoculated (III), thermally treated L3 inoculated (IV), fresh L3 + GO inoculated (V), and a thermally treated L3 + GO inoculated (VI) groups. Rats inoculated with fresh and thermally treated L3 showed abnormal liver and kidney functions associated with the destruction of normal architecture. GO produced a protective effect in rat groups inoculated with L3 extracts + GO via the amelioration of liver and kidney functions, which was confirmed by the marked normal structure on histology. Cooking of L3-infected fish induced severe alterations compared to uncooked fish. The administration of garlic before and after fish eating is recommended to avoid the dangerous effect of anisakids, even if they are cooked.

Acute Anisakiasis: Pharmacological Evaluation of Various Drugs in an Animal Model.

BACKGROUND: The accidental ingestion of the third larval stage of Anisakis can cause acute clinical symptoms, which are relieved via extraction of the larvae. Although this is a highly effective technique, it can only be practiced when the larvae are found in accessible areas of the gastrointestinal tract, and therefore instead the condition has often been treated using various different drugs. AIMS: This study evaluates the effectiveness of gastric acid secretion inhibitors (omeprazole and ranitidine), gastric mucosal protectants (sucralfate) and anthelmintics (mebendazole and flubendazole) in treating anisakiasis in Wistar rats. METHODS: Rats were infected with Anisakis-type I larvae and administered the drugs via a gastric probe. Data were recorded regarding the number of live and dead larvae, their location both within the animal and in its feces, and the presence of gastrointestinal lesions. Additionally, gastric pH was measured and histology performed. RESULTS: While rats in all experimental groups exhibited lesions; those treated with ranitidine and mebendazole showed significantly fewer lesions (50% and 35% of rats exhibited lesions, respectively). Histological examination of the gastric lesions revealed infection-induced changes, but no significant differences were observed between the treated and untreated rats. CONCLUSIONS: Mebendazole was found to be most efficacious in preventing gastrointestinal lesions, followed by ranitidine, which was the most effective antacid of those studied. Both these drugs could thus be considered as part of the conservative management of anisakiasis.

The oil of garlic, Allium sativum L. (Amaryllidaceae), as a potential protectant against Anisakis spp. Type II (L3) (Nematoda) infection in Wistar rats / O óleo de alho, Allium sativum L. (Amaryllidaceae), como potencial protetor contra Anisakis spp. Infecção tipo II (L3) (Nematoda) em ratos Wistar

Abstract The consumption of inadequately thermally treated fish is a public health risk due to the possible propagation of Anisakis larvae. The present study demonstrated the physiological and histopathological changes that accompanied an oral inoculation of crude extracts from fresh and thermally treated Anisakis Type II (L3) in rats. Worms were isolated from a marine fish and examined and identified using light and scanning electron microscopy. The study was performed in 6 rat groups: control (I), garlic oil (GO) inoculated (II), fresh L3 inoculated (III), thermally treated L3 inoculated (IV), fresh L3 + GO inoculated (V), and a thermally treated L3 + GO inoculated (VI) groups. Rats inoculated with fresh and thermally treated L3 showed abnormal liver and kidney functions associated with the destruction of normal architecture. GO produced a protective effect in rat groups inoculated with L3 extracts + GO via the amelioration of liver and kidney functions, which was confirmed by the marked normal structure on histology. Cooking of L3-infected fish induced severe alterations compared to uncooked fish. The administration of garlic before and after fish eating is recommended to avoid the dangerous effect of anisakids, even if they are cooked.

Resumo O consumo de peixe inadequadamente tratado termicamente representa um risco para a saúde pública, com a possibilidade da propagação de larvas de Anisakis. O presente estudo demonstrou as alterações fisiológicas e histopatológicas acompanhadas de inoculação oral de extractos brutos de Anisakis tipo II (L3) frescos e termicamente tratados em ratos. Os vermes foram isolados de um peixe marinho, examinados e identificados por microscopia de luz e eletrônica de varredura. O estudo foi conduzido em 6 grupos de ratos: controle (I), óleo de alho (GO) inoculado (II), L3 fresco inoculado (III), L3 tratado termicamente inoculado (IV), L3 fresco + GO inoculado (V), e um grupo L3 + GO tratado termicamente inoculado (VI). Observou-se que ratos inoculados com L3 fresco e tratados termicamente mostraram funções hepáticas e renais anormais, associadas à destruição da sua arquitetura normal. GO produziu um efeito protector em grupos de ratos inoculados com extractos L3 + GO através da melhoria das funções do fígado e dos rins, o que foi confirmado pela estrutura normal marcada da sua histologia. A cozedura de peixes infectados com L3 induziu alterações mais graves do que os peixes não cozidos. Recomenda-se a administração de alho antes e depois do consumo de peixe, para evitar o efeito perigoso dos anisakids, mesmo que sejam cozidos.

Comparative Proteomics Analysis of Anisakis simplex s.s.-Evaluation of the Response of Invasive Larvae to Ivermectin.

Ivermectin (IVM), an antiparasitic drug, has a positive effect against Anisakis simplex s.s. infection and has been used for the treatment and prevention of anisakiasis in humans. However, the molecular mechanism of action of IVM on A. simplex s.s. remains unknown. Herein, tandem mass tag (TMT) labeling and extensive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis were used to identify the effect of IVM on the proteome of A. simplex s.s. in vitro. During the study, 3433 proteins, of which 1247 had at least two protein unique peptides, were identified. Comparative proteomics analysis revealed that 59 proteins were differentially regulated (DRPs) in IVM-treated larvae, of which 14 proteins were upregulated and 38 were downregulated after 12 h of culture, but after 24 h, 12 proteins were upregulated and 22 were downregulated. The transcription level of five randomly selected DRPs was determined by real-time PCR as a supplement to the proteomic data. The functional enrichment analysis showed that most of the DRPs were involved in oxidoreductase activity, immunogenicity, protein degradation, and other biological processes. This study has, for the first time, provided comprehensive proteomics data on A. simplex s.s. response to IVM and might deliver new insight into the molecular mechanism by which IVM acts on invasive larvae of A. simplex s.s.

Gastro-allergic anisakiasis: The first case reported in Colombia and a literature review / Anisakiasis gastro-alérgica, primera descripción de un caso en Colombia y revisión bibliográfica

Anisakiasis is a zoonotic parasitic disease caused by consumption of raw or undercooked fish or seafood infected with nematodes of the Anisakis, Pseudoterranova or Contracaecum genera. Here, we describe the first case of anisakiasis in Colombia and summarize the available literature. A 52-year-old female with a history of abrupt-onset sharp epigastric pain, nausea, vomit, diarrhea, and urticaria following fish consumption consulted the health service. The physical examination revealed moderate tenderness of the epigastric region; the laboratory evaluation showed leukocytosis and a simple X-ray and ECG showed no abnormalities. The diagnosis was made by endoscopic examination, which revealed a thickened gastric wall and a moving larval worm. An Anisakis larva was found and extracted endoscopically, which relieved the pain of the patient. Clinically, anisakiasis may present as a gastric, intestinal, extragastrointestinal or allergic disease. Diagnosis and treatment of anisakiasis are made by a dietary history, direct visualization and endoscopic extraction of possible larvae, which is the only effective therapy.

La anisakiasis es una enfermedad parasitaria zoonótica causada por el consumo de pescados o mariscos crudos o poco cocidos infectados con nematodos de los géneros Anisakis, Pseudoterranova y Contracaecum. Se describe el primer caso de anisakiasis en Colombia y se resume la literatura médica disponible. Una mujer de 52 años de edad consultó por dolor epigástrico agudo de inicio abrupto, náuseas, vómitos, diarrea y urticaria después de consumir pescado. El examen físico reveló sensibilidad moderada en el epigastrio. El examen de laboratorio evidenció leucocitosis, en tanto que la radiografía simple y el electrocardiograma no reflejaron ninguna anormalidad. El diagnóstico se hizo mediante una endoscopia de vías digestivas altas, la cual reveló engrosamiento de la pared gástrica y un parásito en movimiento. Se encontró una larva de Anisakis y se la extrajo por endoscopia, lo que alivió el dolor de la paciente. Clínicamente, la anisakiasis puede presentarse como una enfermedad gástrica, intestinal, en otros sistemas o alérgica. El diagnóstico se hace con base en la elaboración del historial alimentario del paciente y la visualización directa de las larvas; el único tratamiento efectivo consiste en su extracción endoscópica.

Anisakiasis gastro-alérgica, primera descripción de un caso en Colombia y revisión bibliográfica / Gastro-allergic anisakiasis: The first case reported in Colombia and a literature review

Resumen La anisakiasis es una enfermedad parasitaria zoonótica causada por el consumo de pescados o mariscos crudos o poco cocidos infectados con nematodos de los géneros Anisakis, Pseudoterranova y Contracaecum. Se describe el primer caso de anisakiasis en Colombia y se resume la literatura médica disponible. Una mujer de 52 años de edad consultó por dolor epigástrico agudo de inicio abrupto, náuseas, vómitos, diarrea y urticaria después de consumir pescado. El examen físico reveló sensibilidad moderada en el epigastrio. El examen de laboratorio evidenció leucocitosis, en tanto que la radiografía simple y el electrocardiograma no reflejaron ninguna anormalidad. El diagnóstico se hizo mediante una endoscopia de vías digestivas altas, la cual reveló engrosamiento de la pared gástrica y un parásito en movimiento. Se encontró una larva de Anisakis y se la extrajo por endoscopia, lo que alivió el dolor de la paciente. Clínicamente, la anisakiasis puede presentarse como una enfermedad gástrica, intestinal, en otros sistemas o alérgica. El diagnóstico se hace con base en la elaboración del historial alimentario del paciente y la visualización directa de las larvas; el único tratamiento efectivo consiste en su extracción endoscópica.

Abstract Anisakiasis is a zoonotic parasitic disease caused by consumption of raw or undercooked fish or seafood infected with nematodes of the Anisakis, Pseudoterranova or Contracaecum genera. Here, we describe the first case of anisakiasis in Colombia and summarize the available literature. A 52-year-old female with a history of abrupt-onset sharp epigastric pain, nausea, vomit, diarrhea, and urticaria following fish consumption consulted the health service. The physical examination revealed moderate tenderness of the epigastric region; the laboratory evaluation showed leukocytosis and a simple X-ray and ECG showed no abnormalities. The diagnosis was made by endoscopic examination, which revealed a thickened gastric wall and a moving larval worm. An Anisakis larva was found and extracted endoscopically, which relieved the pain of the patient. Clinically, anisakiasis may present as a gastric, intestinal, extragastrointestinal or allergic disease. Diagnosis and treatment of anisakiasis are made by a dietary history, direct visualization and endoscopic extraction of possible larvae, which is the only effective therapy.

Case Report: A Common Source Outbreak of Anisakidosis in the United States and Postexposure Prophylaxis of Family Collaterals.

We present a case of intussusception with complete small bowel obstruction caused by intestinal anisakidosis requiring surgical resection. A 30-year-old man presented with acute onset of severe abdominal pain 3 days after eating home-cured salmon gravlax. Despite surgery, the patient developed recurrent abdominal pain on two occasions with evidence of continued inflammation proximal to the surgical anastomosis. He was then treated with albendazole and prednisone, and symptoms improved. A decision was made to prophylactically treat two asymptomatic family members who also consumed home-cured gravlax with albendazole, resulting in one individual passing an intact Anisakis worm in her stool. We suggest that albendazole therapy could be considered as a therapy for continued symptoms of anisakidosis and postexposure prophylaxis of Anisakis larvae ingestion from a common source.

Efficiency of Target Larvicides Is Conditioned by ABC-Mediated Transport in the Zoonotic Nematode Anisakis pegreffii.

Anisakiasis is among the most significant emerging foodborne parasitoses contracted through consumption of thermally unprocessed seafood harboring infective Anisakis species larvae. The efficacy of the currently applied anthelminthic therapy in humans and in model organisms has not proven sufficient, so alternative solutions employing natural compounds combined with chemical inhibitors should be explored. By testing toxicity of the natural monoterpenes nerolidol and farnesol and the conventional anthelminthics abamectin and levamisole in the presence/absence of MK-571 and Valspodar, which inhibit the ABC transporter proteins multidrug resistance protein (MRP-like) and P-glycoprotein (P-gp), we determined the preliminary traits of Anisakis detoxifying mechanisms. We found that Anisakis P-gp and MRP-like transporters have a role in the efflux of the tested compounds, which could be useful in the design of novel anthelminthic strategies. As expected, transporter activation and efflux fluctuated over time; they were synchronously active very early postexposure, whereas the activity of one transporter dominated over the other in a time-dependent manner. MRP-like transporters dominated in the efflux of farnesol, and P-gp dominated in efflux of nerolidol, while both were active in effluxing levamisole. The highest toxicity was exerted by abamectin, a P-gp inhibitor per se, which also elicited the highest oxidative stress in treated Anisakis larvae. We suggest that ß-tubulin, observed for the first time as a core element in Anisakis cuticle, might represent an important target for the tested compounds.

Green drugs in the fight against Anisakis simplex-larvicidal activity and acetylcholinesterase inhibition of Origanum compactum essential oil.

Anisakiasis is a fish-borne parasitic disease caused by the consumption of raw or undercooked fish, as well as cephalopods, contaminated by third instar larvae (L3) of species belonging to the genus Anisakis (Anisakidae). Origanum compactum is a small herbaceous aromatic plant endemic to Spain and Morocco. In Morocco, the plant is used under infusion to treat heart diseases and intestinal pains or as preservative for foodstuffs. This is the first time that the O. compactum essential oil is tested against the parasitic nematode Anisakis simplex. The phytochemical analysis by GC-MS revealed carvacrol (50.3%) and thymol (14.8%) as the major oil constituents. The essential oil and its major constituents carvacrol and thymol were tested against A. simplex L3 larvae isolated from blue whiting fish (Micromesistius poutassou). A. simplex mortality (%) after 24 and 48 h of treatment at 1 µl/ml was 100%, with a low LD50 compared with other essential oils and extracts, and the penetration in the agar assay was also reduced, if compared with control wells. The oil, as well as its major constituents, demonstrated a dose-dependent larvicidal activity. Inhibition of the enzyme acetylcholinesterase through a colorimetric assay in 96-well plates was used to elucidate the pharmacological mechanism as this enzyme plays a key role in nematodes neuromuscular function. Interestingly, O. compactum essential oil, carvacrol and thymol inhibited the enzyme, confirming that this could be one of the mechanisms involved in the anthelmintic activity. To the best of our knowledge, this is the first time that O. compactum essential oil is reported as a larvicidal agent against A. simplex L3 larvae.

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

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Direct parasitologial diagnosis of infection with Hysterothylacium aduncum in a patient with epigastralgia.

Anisakis parasitization has been on the rise because some factors, like increased interest in dishes consisting of raw fish. We report a case of epigastralgia with direct diagnosis by endoscopy, which futher study pointed out H. Aduncum as causal agent, a anisakis which is rarely involved in human anisakiasis.

Tumoración mesentérica secundaria a anisakiasis crónica / Mesenteric tumor due to chronic anisakiasis

La anisakiasis intestinal es una enfermedad parasitaria rara y de difícil diagnóstico, ya que las manifestaciones clínicas son inespecíficas, por lo que se considera una enfermedad infradiagnosticada. La sospecha clínica junto a un correcto diagnóstico de la anisakiasis permite la instauración de un tratamiento correcto, ya que en la mayoría de los casos es posible la resolución del cuadro con tratamiento conservador, evitando cirugías innecesarias ante el diagnóstico diferencial preoperatorio de abdomen agudo. Presentamos el caso clínico de un paciente que precisó una intervención quirúrgica urgente secundaria a una reagudización de anisakiasis crónica

Intestinal anisakiasis is a rare parasitic disease and difficult to diagnose due to symptoms are not specific, so it is considered an underdiagnosed disease. The clinical suspicion with a correct diagnosis of anisakiasis allows the establishment of a correct treatment; in most cases, the resolution is possible with conservative treatment, avoiding unnecessary surgery to the preoperative differential diagnosis of acute abdomen. We report the case of a patient who required urgent surgery secondary to an exacerbation of chronic anisakiasis

Peppermint (Mentha piperita) and albendazole against anisakiasis in an animal model.

OBJECTIVES: Therapy against anisakiasis requires invasive techniques to extract L3 , and an effective drug against this nematode is needed. The aim of this study was to determine the efficacy of peppermint essential oil (EO) and its main components against the parasite in comparison to albendazole, a drug currently prescribed to treat anisakiasis. METHODS: We conducted in vitro experiments and studied an experimental model simulating the human infection in Wistar rats. We used polymerase chain reaction restriction fragment length polymorphism to identify A. simplex s.s. and A. pegreffii and determine any differences in their pathogenicity and susceptibility to the treatments. RESULTS: The in vitro and in vivo experiments both showed that the larvicidal activity of peppermint EO, menthol, menthone and menthyl acetate is higher than that of albendazole. Large stomach lesions were observed in 46.7% of the albendazole-treated rats, whereas no gastrointestinal lesions were detected in those treated with peppermint EO, menthol, menthyl acetate or menthone. CONCLUSIONS: In this animal model, treatment with peppermint EO or its main components was more effective than was treatment with albendazole. Lesions were more frequently produced by A. simplex s.s. larvae than by A. pegreffii larvae.

Actividad de albendazol y los aceites esenciales de menta (Mentha piperita) y manzanilla (Matricaria chamomilla) frente Anisakis tipo I / Activity of albendazole and essential oils of peppermint (Mentha piperita) and chamomile (Matricaria chamomilla) against Anisakis type I

Objetivos: Estudiar la posible actividad sinérgica de los aceites esenciales de Mentha piperita y Matricaria chamomilla frente Anisakis tipo I. Material y métodos: Se ensayó tanto in vitro como in vivo, la actividad larvicida de la mezcla de los aceites esenciales utilizando albendazol como fármaco de referencia y midiéndo su eficacia mediante un modelo estadístico. Resultados: En los ensayos in vitro, a las concentraciones de 250 y 187.5 μg/ml, la mortalidad de las larvas fue del 100% mientras que albendazol no mostró eficacia. In vivo, la mezcla de aceites esenciales fue más eficaz, reduciendo las lesiones en ratas en comparación con las tratadas con albendazol y control. Albendazol no redujo de forma significativa el porcentaje de lesiones producidas por las larvas frente al control. Conclusión: La mezcla de aceite esencial de menta y manzanilla podría ser candidata para su uso en el tratamiento/profilaxis de la anisakiasis humana

Aim: We have studied the possible synergistic activity of the essential oils of Mentha piperita and Matricaria chamomile against Anisakis type I. Materials and methods: the larvicidal activity of the mixture of essential oils was tested in vitro and in vivo, using as a reference the drug Albendazole and effectiveness measured through a statistical model. Results: The in vitro and in vivo experiments both evidenced that the larvicidal activity of essential oils, was higher than the Albendazole activity. In the in vitro assay, at concentrations of 250 and 187.5 μg/ml, the mortality was 100% with the mixture of essential oils while albendazole was ineffective at the concentrations studied. In the in vivo assay, the mixture of essential oils, was significantly more effective in the reduction of numbers of lesions with rats in comparison to albendazole treatments and control. Albendazole did not significantly reduce the percentage of lesions caused by larvae vs. control. Conclusion: Considering the results, we have come to the conclusion that the mixture of peppermint essential oils and chamomile could be used in the treatment/prophylaxis of human Anisakis

Activity of Matricaria chamomilla essential oil against anisakiasis.

The increase in diagnosed cases of anisakiasis and the virtual absence of effective treatments have prompted the search for new active compounds against Anisakis L(3) larvae. The biocidal efficacy against different pathogens shown by various essential oils (EO) led us to study the Matricaria chamomilla EO and two of its main components (chamazulene and α-bisabolol) against the L(3) larvae of Anisakis type I. The activity of M. chamomilla EO, chamazulene and α-bisabolol was established by in vitro and in vivo experiments. The EO (125µg/ml) caused the death of all nematodes, which showed cuticle changes and intestinal wall rupture. In the in vivo assays, only 2.2%±1.8 of infected rats treated with M. chamomilla EO showed gastric wall lesions in comparison to 93.3%±3.9 of control. Chamazulene was ineffective, while α-bisabolol showed a high activity to that of the EO in vitro tests but proved less active in vivo. These findings suggest that the larvicidal activity may result from the synergistic action of different compounds of M. chamomilla EO. Neither of the tested products induces irritative damage in the intestinal tissues. In conclusion, M. chamomilla EO is a good candidate for further investigation as a biocidal agent against Anisakis type I.

Two cases of gastric Anisakiasis for which oral administration of a medicine containing wood creosote (Seirogan) was effective.

Anisakiasis is a disease characterized by an abrupt onset of sharp epigastric pain, which occurs typically a few hours after eating raw or undercooked seafood. Anisakiasis was a Japanese localized disease in the past, however has become an illness of concern in many countries where eating Japanese style raw or undercooked seafood has become popular. At present, the only effective treatment is an endoscopic removal of the nematode. Development of an effective medicine is expected. We report two cases of Anisakiasis, the symptoms of which were ameliorated after the administration of an over-the-counter (OTC) medicine containing wood creosote (Seirogan). Also, we examined the in vitro effect of the Seirogan on the viability of the nematode. In the two cases, the strong epigastric pain was subdued promptly after oral intake of the Seirogan. The exposure of Seirogan suppressed the viability of Anisakis Larva in vitro dose dependently. The oral administration of medicine containing wood creosote might be effective as a first aid to ameliorate the symptoms of Anisakiasis.