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.

Mass Spectrometry Based-Proteomic Analysis of Anisakis spp.: A Preliminary Study towards a New Diagnostic Tool.

Anisakiasis is nowadays a well-known infection, mainly caused by the accidental ingestion of Anisakis larvae, following the consumption of raw or undercooked fishes and cephalopods. Due to the similarity of symptoms with those of common gastrointestinal disorders, this infection is often underestimated, and the need for new specific diagnostic tools is becoming crucial. Given the remarkable impact that MALDI-TOF MS biotyping had in the last decade in clinical routine practice for the recognition of bacterial and fungi strains, a similar scenario could be foreseen for the identification of parasites, such as nematodes. In this work, a MALDI-TOF MS profiling of Anisakis proteome was pursued with a view to constructing a first spectral library for the diagnosis of Anisakis infections. At the same time, a shotgun proteomics approach by LC-ESI-MS/MS was performed on the two main fractions obtained from protein extraction, to evaluate the protein species enriched by the protocol. A set of MALDI-TOF MS signals associated with proteins originating in the ribosomal fraction of the nematode extract was selected as a potential diagnostic tool for the identification of Anisakis spp.

Advances in Omic Studies Drive Discoveries in the Biology of Anisakid Nematodes.

Advancements in technologies employed in high-throughput next-generation sequencing (NGS) methods are supporting the spread of studies that, combined with advances in computational biology and bioinformatics, have greatly accelerated discoveries within basic and biomedical research for many parasitic diseases. Here, we review the most updated "omic" studies performed on anisakid nematodes, a family of marine parasites that are causative agents of the fish-borne zoonosis known as anisakiasis or anisakidosis. Few deposited data on Anisakis genomes are so far available, and this still hinders the deep and highly accurate characterization of biological aspects of interest, even as several transcriptomic and proteomic studies are becoming available. These have been aimed at discovering and characterizing molecules specific to peculiar developmental parasitic stages or tissues, as well as transcripts with pathogenic potential as toxins and allergens, with a broad relevance for a better understanding of host-pathogen relationships and for the development of reliable diagnostic tools.

Leukocyte Nucleolus and Anisakis pegreffii-When Falling Apart Means Falling in Place.

The view of the nucleolus as a mere ribosomal factory has been recently expanded, highlighting its essential role in immune and stress-related signalling and orchestrating. It has been shown that the nucleolus structure, formed around nucleolus organiser regions (NORs) and attributed Cajal bodies, is prone to disassembly and reassembly correlated to various physiological and pathological stimuli. To evaluate the effect of parasite stimulus on the structure of the leukocyte nucleolus, we exposed rat peripheral blood mononuclear cells (PBMC) to the crude extract of the nematode A. pegreffii (Anisakidae), and compared the observed changes to the effect of control (RPMI-1640 media), immunosuppressive (MPA) and immunostimulant treatment (bacterial lipopolysaccharide (LPS) and viral analogue polyinosinic:polycytidylic acid (poly I:C)) by confocal microscopy. Poly I:C triggered the most accentuated changes such as nucleolar fragmentation and structural unravelling, LPS induced nucleolus thickening reminiscent of cell activation, while MPA induced disassembly of dense fibrillar and granular components. A. pegreffii crude extract triggered nucleolar segregation, expectedly more enhanced in treatment with a higher dose. This is the first evidence that leukocyte nucleoli already undergo structural changes 12 h post-parasitic stimuli, although these are likely to subside after successful cell activation.

Immunoreactive Proteins in the Esophageal Gland Cells of Anisakis Simplex Sensu Stricto Detected by MALDI-TOF/TOF Analysis.

In plant and animal nematode parasites, proteins derived from esophageal gland cells have been shown to be important in the host-nematodes relationship but little is known about the allergenic potential of these proteins in the genus Anisakis. Taking into account the increase of anisakiasis and allergies related to these nematodes, immunoreactive properties of gland cell proteins were investigated. Two hundred ventricles were manually dissected from L3 stage larvae of Aniskakis simplex s.s. to allow direct protein analysis. Denaturing gel electrophoresis followed by monochromatic silver staining which revealed the presence of differential (enriched) proteins when compared to total nematode extracts. Such comparison was performed by means of 1D and 2D electrophoresis. Pooled antisera from Anisakis spp.-allergic patients were used in western blots revealing the presence of 13 immunoreactive bands in the ventricular extracts in 1D, with 82 spots revealed in 2D. The corresponding protein bands and spots were excised from the silver-stained gel and protein assignation was made by MALDI-TOF/TOF. A total of 13 (including proteoforms) were unambiguously identified. The majority of these proteins are known to be secreted by nematodes into the external environment, of which three are described as being major allergens in other organisms with different phylogenetic origin and one is an Anisakis simplex allergen.

Differences in Gene Expression Profiles of Seven Target Proteins in Third-Stage Larvae of Anisakis simplex (Sensu Stricto) by Sites of Infection in Blue Whiting (Micromesistius poutassou).

The third-stage larvae of the parasitic nematode genus Anisakis tend to encapsulate in different tissues including the musculature of fish. Host tissue penetration and degradation involve both mechanic processes and the production of proteins encoded by an array of genes. Investigating larval gene profiles during the fish infection has relevance in understanding biological traits in the parasite's adaptive ability to cope with the fish hosts' defense responses. The present study aimed to investigate the gene expression levels of some proteins in L3 of A. simplex (s.s.) infecting different tissues of blue whiting Micromesistius poutassou, a common fish host of the parasite in the NE Atlantic. The following genes encoding for Anisakis spp. proteins were studied: Kunitz-type trypsin inhibitor (TI), hemoglobin (hb), glycoprotein (GP), trehalase (treh), zinc metallopeptidase 13 (nas 13), ubiquitin-protein ligase (hyd) and sideroflexin 2 (sfxn 2). Significant differences in gene transcripts (by quantitative real-time PCR, qPCR) were observed in larvae located in various tissues of the fish host, with respect to the control. ANOVA analysis showed that relative gene expression levels of the seven target genes in the larvae are linked to the infection site in the fish host. Genes encoding some of the target proteins seem to be involved in the host tissue migration and survival of the parasite in the hostile target tissues of the fish host.

Comparative Transcriptomics Reveals Clues for Differences in Pathogenicity between Hysterothylacium aduncum, Anisakissimplex sensu stricto and Anisakis pegreffii.

Ascaridoid nematodes are widespread in marine fishes. Despite their major socioeconomic importance, mechanisms associated to the fish-borne zoonotic disease anisakiasis are still obscure. RNA-Seq and de-novo assembly were herein applied to RNA extracted from larvae and dissected pharynx of Hysterothylacium aduncum (HA), a non-pathogenic nematode. Assembled transcripts in HA were annotated and compared to the transcriptomes of the zoonotic species Anisakis simplex sensu stricto (AS) and Anisakis pegreffii (AP). Approximately 60,000,000 single-end reads were generated for HA, AS and AP. Transcripts in HA encoded for 30,254 putative peptides while AS and AP encoded for 20,574 and 20,840 putative peptides, respectively. Differential gene expression analyses yielded 471, 612 and 526 transcripts up regulated in the pharynx of HA, AS and AP. The transcriptomes of larvae and pharynx of HA were enriched in transcripts encoding collagen, peptidases, ribosomal proteins and in heat-shock motifs. Transcripts encoding proteolytic enzymes, anesthetics, inhibitors of primary hemostasis and virulence factors, anticoagulants and immunomodulatory peptides were up-regulated in AS and AP pharynx. This study represents the first transcriptomic characterization of a marine parasitic nematode commonly recovered in fish and probably of negligible concern for public health.

Recent increase of ulcerative lesions caused by Anisakis spp. in cetaceans from the north-east Atlantic.

Species of Anisakis typically infect the stomach of cetaceans worldwide, often causing ulcerative lesions that may compromise the host's health. These nematodes also cause anisakiasis or allergic reactions in humans. To assess the risks of this emerging zoonosis, data on long-term changes in Anisakis infections in cetaceans are necessary. Here, we compare the prevalence and severity of ulcerative lesions caused by Anisakis spp. in five cetacean species stranded along the north-west Spanish coast in 2017-2018 with published data from 1991-1996. Open ulcers were found in 32/43 short-beaked common dolphins, Delphinus delphis; 3/5 striped dolphins, Stenella coeruleoalba; 1/7 bottlenose dolphins, Tursiops truncatus; and 1/3 harbour porpoises, Phocoena phocoena meridionalis; a single individual of long-finned pilot whale, Globicephala melas, was found uninfected. In common dolphins, the mean abundance of open ulcers per host was 1.1 (95% confidence interval: 0.8-1.3), with a maximum diameter (mean ± standard deviation) of 25.4 ± 16.9 mm. Stomachs with scars or extensive fibrosis putatively associated with Anisakis were detected in 14 and five animals, respectively. A molecular analysis based on the mitochondrial cytochrome c oxidase II gene using 18 worms from three cetacean species revealed single or mixed infections of Anisakis simplex sensu stricto and Anisakis pegreffii. Compared with the period 1991-1996, we found a strong increase of prevalence, abundance and extension of ulcerative lesions in most cetacean species. Anisakis populations could have increased in the study area over the last decades, although we cannot rule out that a higher environmental stress has also boosted the pathological effects of these parasites.

Increasing intensities of Anisakis simplex third-stage larvae (L3) in Atlantic salmon of coastal waters of Scotland.

BACKGROUND: Red Vent Syndrome (RVS), a haemorrhagic inflammation of the vent region in Atlantic salmon, is associated with high abundance of Anisakis simplex (s.s.) third-stage larvae (L3) in the vent region. Despite evidence suggesting that increasing A. simplex (s.s.) intensity is a causative factor in RVS aetiology, the definitive cause remains unclear. METHODS: A total of 117 Atlantic salmon were sampled from commercial fisheries on the East, West, and North coasts of Scotland and examined for ascaridoid parasites. Genetic identification of a subsample of Anisakis larvae was performed using the internal transcribed spacer (ITS) region of ribosomal DNA. To assess the extent of differentiation of feeding grounds and dietary composition, stable isotope analysis of carbon and nitrogen was carried out on Atlantic salmon muscle tissue. RESULTS: In the present study, the obtained ITS rDNA sequences matched A. simplex (s.s.) sequences deposited in GenBank at 99-100%. Not all isolated larvae (n = 30,406) were genetically identified. Therefore, the morphotype found in this study is referred to as A. simplex (sensu lato). Anisakis simplex (s.l.) was the most prevalent (100%) nematode with the highest mean intensity (259.9 ± 197.3), in comparison to Hysterothylacium aduncum (66.7%, 6.4 ± 10.2) and Pseudoterranova decipiens (s.l.) (14.5%, 1.4 ± 0.6). The mean intensity of A. simplex (s.l.) represents a four-fold increase compared to published data (63.6 ± 31.9) from salmon captured in Scotland in 2009. Significant positive correlations between A. simplex (s.l.) larvae intensities from the body and the vent suggest that they play a role in the emergence of RVS. The lack of a significant variation in stable isotope ratios of Atlantic salmon indicates that diet or feeding ground are not driving regional differences in A. simplex (s.l.) intensities. CONCLUSIONS: This paper presents the most recent survey for ascaridoid parasites of wild Atlantic salmon from three coastal regions in Scotland. A significant rise in A. simplex (s.l.) intensity could potentially increase both natural mortality rates of Atlantic salmon and possible risks for salmon consumers due to the known zoonotic role of A. simplex (s.s.) and A. pegreffii within the A. simplex (s.l.) species complex.

Seasonal trend of Anisakidae infestation in South Mediterranean bluefish.

A total of 1104 fish samples from markets of Sicily were analysed for the detection and species identification of Anisakidae nematodes. The preliminary analysis of the fish samples showed the presence of 2459 larvae. All the fish species revealed different prevalence of infestation, with a maximum of 100% for Lepidopus caudatus and a minimum of 4.5% in Sardina pilchardus. The 80% of the larvae examined by PCR-RFLP analysis belonged to Anisakis pegreffii species. The seasonal infestation trend of Anisakis was evaluated in all the fish sample examined. The results of the seasonal infestation trend showed a marked connection with the ecological aspects of the fish species examined. As far as we know, this work report for the first time important ecological aspects of Lepidopus caudatus specimens of South Mediterranean. This work could be useful to plan a seasonal fishing strategy aimed at reducing the health risks related to Anisakis.

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.

Functional insights into the infective larval stage of Anisakis simplex s.s., Anisakis pegreffii and their hybrids based on gene expression patterns.

BACKGROUND: Anisakis simplex sensu stricto and Anisakis pegreffii are sibling species of nematodes parasitic on marine mammals. Zoonotic human infection with third stage infective larvae causes anisakiasis, a debilitating and potentially fatal disease. These 2 species show evidence of hybridisation in geographical areas where they are sympatric. How the species and their hybrids differ is still poorly understood. RESULTS: Third stage larvae of Anisakis simplex s.s., Anisakis pegreffii and hybrids were sampled from Merluccius merluccius (Teleosti) hosts captured in waters of the FAO 27 geographical area. Specimens of each species and hybrids were distinguished with a diagnostic genetic marker (ITS). RNA was extracted from pools of 10 individuals of each taxon. Transcriptomes were generated using Illumina RNA-Seq, and assembled de novo. A joint assembly (here called merged transcriptome) of all 3 samples was also generated. The inferred transcript sets were functionally annotated and compared globally and also on subsets of secreted proteins and putative allergen families. While intermediary metabolism appeared to be typical for nematodes in the 3 evaluated taxa, their transcriptomes present strong levels of differential expression and enrichment, mainly of transcripts related to metabolic pathways and gene ontologies associated to energy metabolism and other pathways, with significant presence of excreted/secreted proteins, most of them allergens. The allergome of the 2 species and their hybrids has also been thoroughly studied; at least 74 different allergen families were identified in the transcriptomes. CONCLUSIONS: A. simplex s.s., A. pegreffi and their hybrids differ in gene expression patterns in the L3 stage. Strong parent-of-origin effects were observed: A. pegreffi alleles dominate in the expression patterns of hybrids albeit the latter, and A. pegreffii also display significant differences indicating that hybrids are intermediate biological entities among their parental species, and thus of outstanding interest in the study of speciation in nematodes. Analyses of differential expression based on genes coding for secreted proteins suggests that co-infections presents different repertoires of released protein to the host environment. Both species and their hybrids, share more allergen genes than previously thought and are likely to induce overlapping disease responses.

Tissue-specific transcriptomes of Anisakis simplex (sensu stricto) and Anisakis pegreffii reveal potential molecular mechanisms involved in pathogenicity.

BACKGROUND: Larval stages of the sibling species of parasitic nematodes Anisakis simplex (sensu stricto) (s.s.) (AS) and Anisakis pegreffii (AP) are responsible for a fish-borne zoonosis, known as anisakiasis, that humans aquire via the ingestion of raw or undercooked infected fish or fish-based products. These two species differ in geographical distribution, genetic background and peculiar traits involved in pathogenicity. However, thus far little is known of key molecules potentially involved in host-parasite interactions. Here, high-throughput RNA-Seq and bioinformatics analyses of sequence data were applied to the characterization of the whole sets of transcripts expressed by infective larvae of AS and AP, as well as of their pharyngeal tissues, in a bid to identify transcripts potentially involved in tissue invasion and host-pathogen interplay. RESULTS: Approximately 34,000,000 single-end reads were generated from cDNA libraries for each species. Transcripts identified in AS and AP encoded 19,403 and 10,424 putative peptides, respectively, and were classified based on homology searches, protein motifs, gene ontology and biological pathway mapping. Differential gene expression analysis yielded 226 and 339 transcripts upregulated in the pharyngeal regions of AS and AP, respectively, compared with their corresponding whole-larvae datasets. These included proteolytic enzymes, molecules encoding anesthetics, inhibitors of primary hemostasis and virulence factors, anticoagulants and immunomodulatory peptides. CONCLUSIONS: This work provides the scientific community with a list of key transcripts expressed by AS and AP pharyngeal tissues and corresponding annotation information which represents a ready-to-use resource for future functional studies of biological pathways specifically involved in host-parasite interplay.

Excretory/secretory proteases and mechanical movement of Anisakis pegreffii infective larvae in the penetration of BALB/c mice gastrointestine.

Anisakiasis is a human parasitic disease caused by infection with the infective larvae of Anisakis. Accidental infection in humans causes the gastrointestinal pathophysiological effects of mechanical tissue damage by migrating larvae. The mechanism of the infective larval invasion and migration is suspected to involve larval excretory/secretory proteases and motility. This study demonstrates the penetration rate of the infective larvae of Anisakis pegreffii in mouse gastrointestine depends on the time after infection, and that only 15% of larvae remain in the gastrointestinal tract 3 h after infection. Strong activities of matrix metalloproteinases (MMPs) and serine proteases, especially plasmin, were found in the excretory/secretory products of A. pegreffii; these can be inhibited by ONO-4817 and phenylmethylsulfonyl fluoride, respectively. The protease activity was also significantly decreased in another 1 h of cultivation of larvae in fresh 0.9% normal saline (NS) after previous cultivation for 48 h in NS. The motility scores of larvae were significantly lower after 48 h of cultivation in NS. The penetration rate of A. pegreffii larvae in the gastrointestine of infected mice sequentially were 90% in the freshly prepared, 68% in serine protease inhibited, 55% in MMPs inhibited larvae, and 16% in larvae cultivated in NS for 48 h. Therefore, this study demonstrates that MMPs and serine proteases excreted and secreted by A. pegreffii and the mechanical movement of infective larvae participate in the penetration of the gastrointestine of mice after infection.

Molecular diagnosis of allergy to Anisakis simplex and Gymnorhynchus gigas fish parasites

Background: There has been an increase in the prevalence of hypersensitivity to Anisakis simplex. There are fish parasites other than Anisakis simplex whose allergenicity has not yet been studied. Objective: To assess IgE hypersensitivity caused by fish parasite allergens in patients with gastro-allergic symptoms after consumption of fish, shellfish or cephalopods, compared with healthy subjects, pollen allergic individuals and children with digestive symptoms after eating marine food. Methods: We carried out in vivo tests (skin prick) and in vitro tests (specific IgE determination, Western blot) and component resolved diagnostics (CRD) using microarray analysis in all patients. Results: CRD better detected sensitisation to allergens from marine parasites than skin prick tests and determination of specific IgE by CAP. Sensitisation to Gymnorhynchus gigas was detected in 26% of patients measured by skin prick tests and 36% measured by IgE. Conclusions: The prevalence of hypersensitivity to marine parasite allergens other than Anisakis simplex should be studied, and the most appropriate technique for this is CRD (AU)

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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.

Anisakis y pólipo en colon, una asociación infrecuente / Anisakis and colonic polyp, a rare association

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El anisakis y sus enfermedades como enfermedad profesional / Anisakis and disease as an occupational disease

El anisakis es un parásito complejo de los animales marinos. El hombre no es el hospedador adecuado, ya que en él la larva L3 no puede completar su ciclo vital, siendo por tanto un hospedador accidental u ocasional, que se interpone en el ciclo vital del nemátodo, y al que el anisakis le puede causar tanto la parasitación como la alergia, cuando ingiere pescado o cefalópodos crudos o poco cocinados: tales como sushi, sashimi, ceviche gravlax noruego, lomi-lomi hawaiano, así como pescados en escabeche, vinagre, desecado, ahumados, semiconservas, o en salazón que contengan este parásito en su forma de larva en estadio tres L3. Las enfermedades por anisakis constituyen un problema de salud pública, generalizado y que ha aumentado su incidencia en los últimos años. En 2012, se notificaron 20.000 casos de anisakiasis en todo el mundo. España es el segundo país con mayor número de intoxicaciones por Anisakis después de Japón. Diversos estudios en España sobre la prevalencia de la sensibilización reflejan una elevada prevalencia entre pacientes que han sufrido episodios de reacciones alérgicas. El 50 por ciento de quienes manipulan directamente el pescado presentan sensibilidad a anisakis. Siendo por tanto cada vez más común la afectación por y en el desarrollo del trabajo, constituyendo entonces procesos considerados como enfermedad profesional cuando se dan las circunstancias determinantes causales de trabajo, riesgo y exposición que para ello deben concurrir. La afección puede presentarse tanto por contacto laboral como por contacto en el consumo ordinario de pescado infestado, lo que hará difícil el estimar como causa exclusiva o consecuente la aparición de la enfermedad a consecuencia del desempeño del trabajo. Los sectores laborales de riesgo por enfermedades por anisakis, son los pescadores, pescaderos («pescateros») y cocineros. En general todos aquellos que trabajan tanto en la captura, limpieza, manipulación, venta, elaboración o preparación del pescado contaminado. Describimos en este artículo las enfermedades por anisakis, los trabajos de riesgo, las características que debieran reunir para ser enfermedad profesional y dos casos ilustrativos cuya consideración lo fue como esta contingencia laboral. Conclusión: Las enfermedades por anisakis y su consideración como enfermedad profesional, viene sujeta a la propia definición de lo que se considera así en nuestra norma legal, es decir que sea una enfermedad contraída como consecuencia del trabajo, en las actividades que se describen en el cuadro de enfermedades profesionales y por la acción del agente o sus sustancias que se recogen en el anexo I del actual cuadro. Además para que se admita como enfermedad profesional debemos de tener un diagnóstico firme (encuadrado en el cuadro de enfermedad profesional), un agente causal (recogido en el listado) y una exposición al anisakis desarrollada en el ejercicio del trabajo (actividades recogidas en el listado). El listado o cuadro de enfermedad profesional es un sistema cerrado para agentes causales y enfermedades causadas, y la adscripción no es fácil. La parasitación tendría cabida en el grupo de enfermedades por agentes biológicos, la alergia podría discutirse su adscripción pero suele recogerse también en este grupo, el asma lo recogeríamos en el grupo de enfermedades respiratorias (causadas por inhalación dice el texto legal) y las manifestaciones dérmicas en el grupo de las enfermedades profesionales de la piel. La exposición laboral al anisakis, es necesaria e imprescindible para que el proceso sea calificado de enfermedad profesional, pero esta exposición no siempre estará bien delimitada, y sin embargo debe estar probada. Puede haber existido una sensibilización fuera del trabajo y una aparición o desarrollo de la enfermedad por reiterada exposición o contacto con el anisakis en el trabajo lo que dificultará su calificación ocupacional

Anisakis is a complex parasite found in marine animals. The human being is not the appropriate host, since the L3 larvae cannot complete its life cycle from within, thus making it an accidental or occasional host that stands in the life cycle of the nematode. The anisakis can cause both parasitism and allergy when the following infected food is eaten: raw fish or cephalopods, undercooked sushi, sashimi, Norwegian gravlax ceviche, Hawaiian lomi-lomi and fish in brine, vinegar, dried, smoked, semi-conserve, or salted. They might contain this parasite in the third-stage larvae form, L3. Anisakis diseases constitute a problem for the public health and its incidence has increased in recent years. In 2012, 20,000 anisakis cases were reported worldwide. Spain is the second country with the highest number of poisoning due to anisakis after Japan. Several studies in Spain on the prevalence of sensitization reflect a high prevalence among patients who have suffered episodes of allergic reactions. 50% of the people who are directly handling fish present anisakis sensitivity. Thus being increasingly affected by common development and work, turning out in processes considered as an occupational disease where the determining circumstances are grounds for work, risk and exposure that it should attend. The condition may occur both work contact and contact in the regular consumption of infested fish, which will make it difficult to estimate as exclusive or consequential onset of the disease as a result of job performance cause. Labor sectors anisakis disease risk are fishermen, fishmongers and cooks. In general, all those who work both in the capture, cleaning, handling, sale, processing or preparation of contaminated fish. We describe in this article anisakis diseases, risk jobs and the characteristics that should gather for occupational disease and two illustrative cases whose consideration was such this labour contingency. Conclusions: Anisakis diseases and its consideration as an occupational disease is subject to the definition of what is considered in our legal norm in Spain, which means that it is a disease contracted as a result of work activities described in the box of occupational diseases and the action of the agents or substances listed in Annex I of the current frame diseases. In addition, in order to be recognised as an occupational disease we must have a firm diagnosis (Table occupational disease), a causative agent (referred to in the list) and an exposure to anisakis developed in the course of work (activities included in the list). The list box or occupational disease frame is a closed causal agent and caused diseases, and the attachment is not easy. Parasitism would fit in the group of diseases by biological agents; allergy disease could discuss its assignment but it has also usually been included in this group; asthma would collect in the group of respiratory diseases (caused by inhalation according to the legal text) and dermal manifestations in the group of occupational skin diseases. Occupational exposure to anisakis is necessary and essential for the process of being qualified as occupational disease, and although this exhibition is not always well defined must be tested. There may have been a sensitization out of work and an onset or development of the disease by repeated exposure or contact with the anisakis at work which would complicate its occupational qualification

Anisakiasis Causing Acute Dysentery in Malaysia.

Human anisakiasis is a zoonosis acquired by eating raw or undercooked infected seafood. Herein, we report a case of acute dysentery caused by anisakiasis in a 64-year-old man in Malaysia. A colonoscopy was performed and a nematode larva was found penetrating the mucosa of the ascending colon. Bleeding was observed at the site of penetration. Y-shaped lateral epidermal cords were seen from the cross section of the worm, which is a prominent feature of Anisakis larva. Molecular analysis using polymerase chain reaction of cytochrome oxidase 2 (cox2) gene confirmed the specimen to be larva of Anisakis simplex.

Infección por Anisakis con presentación atípica: a propósito de un caso / Anisakis infection with atypical presentation: Report of a case

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