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Comparative Proteomics Analysis of s.s.-Evaluation of the Response of Invasive Larvae to Ivermectin.Genes Jun 2020Ivermectin (IVM), an antiparasitic drug, has a positive effect against s.s. infection and has been used for the treatment and prevention of anisakiasis in humans....
Ivermectin (IVM), an antiparasitic drug, has a positive effect against 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 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 s.s. . 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 s.s. response to IVM and might deliver new insight into the molecular mechanism by which IVM acts on invasive larvae of s.s.
Topics: Animals; Anisakiasis; Anisakis; Chromatography, Liquid; Gene Expression Regulation; Humans; Ivermectin; Larva; Proteome; Proteomics; Tandem Mass Spectrometry
PubMed: 32604878
DOI: 10.3390/genes11060710 -
Emerging Infectious Diseases Jun 2022Human Pseudoterranova decipiens larval infections were diagnosed by molecular analysis of mitochondrial cox1 and nd1 genes in 12 health check-up patients in South Korea...
Human Pseudoterranova decipiens larval infections were diagnosed by molecular analysis of mitochondrial cox1 and nd1 genes in 12 health check-up patients in South Korea during 2002-2020. Based on high genetic identity (99.3%-100% for cox1 and 96.7%-98.0% for nd1), we identified all 12 larvae as P. decipiens sensu stricto.
Topics: Animals; Anisakiasis; Anisakis; Ascaridoidea; Humans; Larva; Republic of Korea
PubMed: 35608950
DOI: 10.3201/eid2806.212483 -
Scientific Reports Jul 2020The high frequency of infection by Anisakis simplex (A. simplex) has led to an increase in IgE sensitization, turning allergy to this parasite a relevant contemporary...
The high frequency of infection by Anisakis simplex (A. simplex) has led to an increase in IgE sensitization, turning allergy to this parasite a relevant contemporary health problem. Improving the lack of conventional diagnosis test specificity is crucial to better understand these clinical scenarios. Specific IgE (sIgE) to A. simplex extract by ImmunoCAP (Anisakis-sIgE) was determined in sera from 403 blood donors (BD) from Cantabria (North of Spain) of which 51 subjects resulted sensitized. Among these latter, 47 were asymptomatic (sABD). The values of total IgE, prick-test, Anisakis-sIgE, and sIgE to Ani s 1 (anti-rAni s 1) and Ani s 7 (anti-rAni s 7) were compared between 46 sABD and 49 A. simplex allergic patients. The IgE seroprevalence by ImmunoCAP among BD was 12.65%. Allergic patients and sABD showed significant differences in all serum biomarkers evaluated. The area under the curve was assessed for Anisakis-sIgE (0.892), sIgE-rAni s 1 (0.672) and sIgE-rAni s 7 (0.668). After a severe reaction, significantly higher levels of Anisakis-sIgE and sIgE anti-rAni s 1 were detected. Determinations of sIgE by ImmunoCAP, Ani s 1 and Ani s 7 presented different sensitization patterns between allergic and asymptomatic individuals. The Ani s 1 allergen arises as a possible biomarker to detect patients at risk of suffering severe allergic reactions.
Topics: Adult; Aged; Allergens; Animals; Anisakiasis; Anisakis; Antigens, Helminth; Biomarkers; Calcium-Binding Proteins; Cross-Sectional Studies; Dermatophagoides pteronyssinus; Enzyme-Linked Immunosorbent Assay; Female; Helminth Proteins; Humans; Hypersensitivity; Immunoglobulin E; Male; Middle Aged; Penaeidae; Prevalence; Prospective Studies; ROC Curve; Seroepidemiologic Studies
PubMed: 32647149
DOI: 10.1038/s41598-020-67786-w -
Genes Jun 2020In 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...
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 . 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 . 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 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 allergen.
Topics: Allergens; Animals; Anisakiasis; Anisakis; Esophagus; Fish Diseases; Host-Parasite Interactions; Humans; Larva; Phylogeny; Proteins; Seafood; Species Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 32580523
DOI: 10.3390/genes11060683 -
Biology Feb 2021( infection, in humans, causes a series of clinical manifestations affecting the gastro-intestinal tract known as Anisakiasis/Anisakidosis. Patients may also present...
BACKGROUND
( infection, in humans, causes a series of clinical manifestations affecting the gastro-intestinal tract known as Anisakiasis/Anisakidosis. Patients may also present allergic manifestations such as hives and/or angioedema and even anaphylactic shock. The aim of this study was to investigate whether aquacultured fish could be considered -free food and constitute a safe, alternative, wild-capture fish food for Gastro-Allergic Anisakiasis (GAA)-sensitized subjects.
METHODS
Protein extracts from larvae in the third stage (L3) and from edible part of heavily infected horse mackerel () and aquacultured sea bream, have been tested for allergens presence by immunological analysis. Western blot analysis using, as source of specific allergens antibodies, serum samples from subjects referring allergic symptoms after raw fish ingestion, was performed. These subjects showed high levels of specific IgE anti allergens determined by clinical laboratory tests (ISAC test).
RESULTS
Our data demonstrate the presence of Ani s4 allergen in both infected and aquacultured fish extracts, providing a possible interpretation for the allergic manifestations reported by subjects, already sensitized to , who ate frozen or well-cooked or, even, aquacultured fish.
CONCLUSIONS
The present data stimulate more accurate prophylaxis suggestions for allergy and more specific controls of fishmeal used in aquaculture.
PubMed: 33540832
DOI: 10.3390/biology10020106 -
Scientific Reports Oct 2020An amendment to this paper has been published and can be accessed via a link at the top of the paper.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
PubMed: 33110175
DOI: 10.1038/s41598-020-75954-1 -
Journal of Parasitic Diseases :... Sep 2020Anisakidosis is a zoonotic infection caused by members of the family Anisakidae. The presence of anisakid larvae in fish poses risk for humans and dissuade consumers...
Anisakidosis is a zoonotic infection caused by members of the family Anisakidae. The presence of anisakid larvae in fish poses risk for humans and dissuade consumers from purchasing infected products. Although fish constitute important component of Egyptian diet, the prevalence of anisakid larvae in marketed fish in Egypt is not well described. Furthermore, the species of anisakid larvae is not defined in most of the available studies due to the over reliance on morphological analyses. The aim of the current work was to assess the prevalence and intensity of anisakid larvae in three common marketed fish in Egypt (Atlantic herring, Mediterranean horse mackerel and Atlantic mackerel) and to determine the species of the isolated larvae using morphological and molecular methods. Light and scanning electron microscope (SEM) analyses revealed the details of the isolated larvae. However, partial sequencing of cytochrome oxidase subunite-1 () gene revealed that all larvae isolated from Atlantic herring and Mediterranean horse mackerel belonged to sensu stricto with prevalence of 87.1% and 83.3%, respectively, whereas Atlantic mackerel harbored with a prevalence of 42.8%. The Mediterranean horse mackerel demonstrated the highest larval mean intensity (n = 20 larvae/infected fish). This study highlights the importance of these fish as potential reservoirs for human anisakiasis in Egypt and possibly in other coastal countries.
PubMed: 32801505
DOI: 10.1007/s12639-020-01222-8 -
Cells Nov 2019Excretory and secretory products are crucial for parasite infectivity and host immunomodulation, but the functioning and ultrastructure of the excretory gland cell (EC)...
Excretory and secretory products are crucial for parasite infectivity and host immunomodulation, but the functioning and ultrastructure of the excretory gland cell (EC) that produces these products are still scarcely understood and described. In light of growing reports on anisakiasis cases in Europe, we aimed to characterise the EC of larval and adult . In the latter, EC starts 0.85 mm from the head tip, measuring 1.936 × 0.564 mm. Larval EC shows a long nucleus with thorn-like extravaginations toward the cytoplasm, numerous electron-dense and -lucent secretory granules spanning from the perinuclear to subplasmalemmal space, an elevated number of free ribosomes, small, spherical mitochondria with few cristae and a laminated matrix, small and few Golgi apparatuses, and few endoplasmic reticula, with wide cisternae complexes. Ultrastructure suggests that anaerobic glycolysis is the main metabolic pathway, obtained through nutrient endocytosis across the pseudocoelomic surface of the EC plasmalemma and its endocytic canaliculi. Thorn-like extravaginations of EC karyotheca likely mediate specific processes (Ca signaling, gene expression, transport, nuclear lipid metabolism) into the extremely wide EC cytosol, enabling focal delivery of a signal to specific sites in a short time. These functional annotations of parasitic EC should help to clarify anisakiasis pathogenesis.
Topics: Anaerobiosis; Animals; Ascaridoidea; Exocrine Glands; Glycolysis; Larva; Microscopy, Confocal; X-Ray Microtomography
PubMed: 31744245
DOI: 10.3390/cells8111451 -
Genomics Sep 2021Parasitism is a highly successful life strategy and a driving force in genetic diversity that has evolved many times over. Accidental infections of non-targeted hosts...
Parasitism is a highly successful life strategy and a driving force in genetic diversity that has evolved many times over. Accidental infections of non-targeted hosts represent an opportunity for lateral host switches and parasite niche expansion. However, if directed toward organisms that are phylogenetically distant from parasite's natural host, such as humans, it may present a dead-end environment where the parasite fails to mature or is even killed by host immunity. One example are nematodes of Anisakidae family, genus Anisakis, that through evolution have lost the ability to propagate in terrestrial hosts, but can survive for a limited time in humans causing anisakiasis. To scrutinize versatility of Anisakis to infect an evolutionary-distant host, we performed transcriptomic profiling of larvae successfully migrating through the rat, a representative model of accidental human infection and compared it to that of larvae infecting an evolutionary-familiar, paratenic host (fish). In a homeothermic accidental host Anisakis upregulated ribosome-related genes, cell division, cuticle constituents, oxidative phosphorylation, in an unsuccessful attempt to molt to the next stage. In contrast, in the paratenic poikilothermic host where metabolic pathways were moderately upregulated or silenced, larvae prepared for dormancy by triggering autophagy and longevity pathways. Identified differences and the modelling of handful of shared transcripts, provide the first insights into evolution of larval nematode virulence, warranting their further investigation as potential drug therapy targets.
Topics: Animals; Anisakiasis; Anisakis; Fishes; Larva; Rats; Virulence Factors
PubMed: 34186188
DOI: 10.1016/j.ygeno.2021.06.032 -
Chemical & Pharmaceutical Bulletin 2020Anisakiasis is common in countries where raw or incompletely cooked marine fish are consumed. Currently, effective therapeutic methods to treat anisakiasis are...
Anisakiasis is common in countries where raw or incompletely cooked marine fish are consumed. Currently, effective therapeutic methods to treat anisakiasis are unavailable. A recent study found that wood creosote inactivates the movement of Anisakis species. Essential oil of Origanum compactum containing carvacrol and thymol, which are similar to the constituents of wood creosote, was reported to inactivate Anisakis by inhibiting its acetylcholinesterase. We examined whether wood creosote can also inhibit acetylcholinesterase. We examined the effect of components of wood creosote using the same experimental method. A computer simulation experiment (molecular docking) was also performed. Here, we demonstrate that wood creosote inactivated acetylcholinesterase in a dose-dependent manner with an IC of 0.25 mg/mL. Components of wood creosote were also tested individually: 5-methylguaiacol, p-cresol, guaiacol, o-cresol, 2,4-dimethylphenol, m-cresol, phenol and 4-methylguaiacol inactivated the enzyme with an IC of 14.0, 5.6, 17.0, 6.3, 3.9, 10.0, 15.2 and 27.2 mM, respectively. The mechanism of acetylcholinesterase inactivation was analyzed using a computer-based molecular docking simulation, which employed a three-dimensional structure of acetylcholinesterase and above phenolic compounds as docking ligands. The simulation indicated that the phenolic compounds bind to the active site of the enzyme, thereby competitively blocking entry of the substrate acetylcholine. These findings suggest that the mechanism for the inactivation of Anisakis movement by wood creosote is due to inhibition of acetylcholinesterase needed for motor neuron activity.
Topics: Acetylcholinesterase; Animals; Cholinesterase Inhibitors; Creosote; Dose-Response Relationship, Drug; Electrophorus; Molecular Docking Simulation; Molecular Structure; Phenols; Structure-Activity Relationship; Wood
PubMed: 33268651
DOI: 10.1248/cpb.c20-00583