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International Journal of Molecular... Apr 2022s. s. is a parasitic nematode of marine mammals and causative agent of anisakiasis in humans. The cuticle and intestine of the larvae are the tissues most responsible...
Tandem Mass Tagging (TMT) Reveals Tissue-Specific Proteome of L4 Larvae of s. s.: Enzymes of Energy and/or Carbohydrate Metabolism as Potential Drug Targets in Anisakiasis.
s. s. is a parasitic nematode of marine mammals and causative agent of anisakiasis in humans. The cuticle and intestine of the larvae are the tissues most responsible for direct and indirect contact, respectively, of the parasite with the host. At the L4 larval stage, tissues, such as the cuticle and intestine, are fully developed and functional, in contrast to the L3 stage. As such, this work provides for the first time the tissue-specific proteome of s. s. larvae in the L4 stage. Statistical analysis (FC ≥ 2; -value ≤ 0.01) showed that 107 proteins were differentially regulated (DRPs) between the cuticle and the rest of the larval body. In the comparison between the intestine and the rest of the larval body at the L4 stage, 123 proteins were identified as DRPs. Comparison of the individual tissues examined revealed a total of 272 DRPs, with 133 proteins more abundant in the cuticle and 139 proteins more abundant in the intestine. Detailed functional analysis of the identified proteins was performed using bioinformatics tools. Glycolysis and the tricarboxylic acid cycle were the most enriched metabolic pathways by cuticular and intestinal proteins, respectively, in the L4 stage of s. s. The presence of two proteins, folliculin (FLCN) and oxoglutarate dehydrogenase (OGDH), was confirmed by Western blot, and their tertiary structure was predicted and compared with other species. In addition, host-pathogen interactions were identified, and potential new allergens were predicted. The result of this manuscript shows the largest number of protein identifications to our knowledge using proteomics tools for different tissues of L4 larvae of s. s. The identified tissue-specific proteins could serve as targets for new drugs against anisakiasis.
Topics: Animals; Anisakiasis; Anisakis; Carbohydrate Metabolism; Humans; Larva; Mammals; Proteome
PubMed: 35457153
DOI: 10.3390/ijms23084336 -
Materials Today. Bio Jun 2022Engineering the surfaces of biological organisms allows the introduction of novel functions and enhances their native functions. However, studies on surface engineering...
Engineering the surfaces of biological organisms allows the introduction of novel functions and enhances their native functions. However, studies on surface engineering remained limited to unicellular organisms. Herein, nematode surfaces are engineered through hydrogelation mediated by horseradish peroxidase (HRP) anchored to nematode cuticles. With this method, hydrogel sheaths of approximately 10-μm thickness are fabricated from a variety of polysaccharides, proteins, and synthetic polymers. and coated with a hydrogel sheath showed a negligible decrease in viability, chemotaxis and locomotion. Hydrogel sheaths containing UV-absorbable groups and catalase functioned as shields to protect nematodes from UV and hydrogen peroxide, respectively. The results also showed that hydrogel sheaths containing glucose oxidase have the potential to be used as living drug delivery systems for cancer therapy. The nematode functionalization method developed in this study has the potential to impact a wide range of fields from agriculture to medicine.
PubMed: 35774197
DOI: 10.1016/j.mtbio.2022.100328 -
Biology Open Mar 2019L3 larvae infect fish and other seafood species such as squid or octopi; therefore, humans consuming raw or undercooked fish may become accidental hosts for this...
L3 larvae infect fish and other seafood species such as squid or octopi; therefore, humans consuming raw or undercooked fish may become accidental hosts for this parasite. These larvae are induced to enter hypometabolism by cold temperatures. It is assumed that sugars (in particular trehalose and glycogen) are instrumental for survival under environmental stress conditions. To elucidate the mechanisms of environmental stress response in , we observed the effects of starvation and temperature on trehalose and glycogen content, the activity of enzymes metabolizing those sugars, and the relative expression of genes of trehalose and glycogen metabolic pathways. The L3 of synthesize trehalose both in low (0°C) and high temperatures (45°C). The highest content of glycogen was observed at 45°C at 36 h of incubation. On the second day of incubation, tissue content of trehalose depended on the activity of the enzymes: TPS was more active at 45°C, and TPP was more active at 0°C. The changes in TPP activity were consistent with the transcript level changes of the TPP gene, and the trehalose level, while glycogen synthesis correlates with the expression of glycogen synthase gene at 45°C; this suggests that the synthesis of trehalose is more essential. These results show that trehalose plays a key role in providing energy during the thermotolerance and starvation processes through the molecular and biochemical regulation of trehalose and glycogen metabolism.
PubMed: 30824422
DOI: 10.1242/bio.040014 -
Genes Aug 2020The total proteomes of s.s., and their hybrid genotype have been compared by quantitative proteomics (iTRAQ approach), which considers the level of expressed proteins....
The total proteomes of s.s., and their hybrid genotype have been compared by quantitative proteomics (iTRAQ approach), which considers the level of expressed proteins. Comparison was made by means of two independent experiments considering four biological replicates of and two each for and hybrid between both species. A total of 1811 and 1976 proteins have been respectively identified in the experiments using public databases. One hundred ninety-six proteins were found significantly differentially expressed, and their relationships with the nematodes' biological replicates were estimated by a multidimensional statistical approach. Results of pairwise Log ratio comparisons among them were statistically treated and supported in order to convert them into discrete character states. Principal component analysis (PCA) confirms the validity of the method. This comparison selected thirty seven proteins as discriminant taxonomic biomarkers among , and their hybrid genotype; 19 of these biomarkers, encoded by ten loci, are specific allergens of (Ani s7, Ani s8, Ani s12, and Ani s14) and other (Ancylostoma secreted) is a common nematodes venom allergen. The rest of the markers comprise four unknown or non-characterized proteins; five different proteins (leucine) related to innate immunity, four proteolytic proteins (metalloendopeptidases), a lipase, a mitochondrial translocase protein, a neurotransmitter, a thyroxine transporter, and a structural collagen protein. The proposed methodology (proteomics and statistical) solidly characterize a set of proteins that are susceptible to take advantage of the new targeted proteomics.
Topics: Animals; Anisakis; Biomarkers; Chromatography, Liquid; DNA Barcoding, Taxonomic; Gene Regulatory Networks; Genotype; Hybridization, Genetic; Mass Spectrometry; Proteome; Proteomics
PubMed: 32785065
DOI: 10.3390/genes11080913 -
Pathogens (Basel, Switzerland) Sep 2020The third-stage larvae (L3) of are the most important source of hidden allergens in seafood products. However, there exist no commercial methods for detecting proteins...
The third-stage larvae (L3) of are the most important source of hidden allergens in seafood products. However, there exist no commercial methods for detecting proteins in food. Furthermore, only a few methods have been validated for the detection of in thermally processed food. The aims of our study are (i) the development and validation of high-sensitivity chemiluminescent (CL) immunoassays for the detection of proteins in processed seafood, (ii) and antigen detection in common seafood products from Polish markets. We developed and validated CL sandwich ELISA (S-ELISA) and CL competitive ELISA (C-ELISA) methods for proteins detection in food, with respective detection limits of 0.5 and 5 ng/mL. The usefulness of the assays for detecting proteins in highly processed food was evaluated by examination of autoclaved canned fish spiked with larvae (1-8 larvae/200 g). Commercial real-time PCR was unable to detect in autoclaved samples at all levels of enrichment with larvae. CL-S-ELISA was used to test various types of seafood products from Polish markets. Among all tested products ( = 259), 28% were positive. antigens were found mostly ( = 39) in smoked fish products: mackerel, herring, cod, and hake. Other positive samples were found in marinated herrings, canned cod livers, canned mackerels, and surimi sticks. In tuna, Atlantic argentine, anchovy, sardine, sprat, and squid products, antigens were not detected. This study provides novel effective tools for the detection of proteins in processed food and highlights the potential allergic hazards for -sensitized Polish consumers of seafood.
PubMed: 32977528
DOI: 10.3390/pathogens9100777 -
Scientific Reports Mar 2022The family Anisakidae, mainly represented by Anisakis simplex s.l. and Pseudoterranova decipiens, encompasses zoonotic nematodes infecting many marine fish. Both are...
The family Anisakidae, mainly represented by Anisakis simplex s.l. and Pseudoterranova decipiens, encompasses zoonotic nematodes infecting many marine fish. Both are responsible for gastrointestinal disease in humans after ingestion of a live larva by consumption of undercooked fish, and, in the case of A. simplex, an allergic reaction may occur after consuming or even handling infected fish. Due to its phylogenetic relatedness with A. simplex, few studies investigated the allergenic potential of P. decipiens, yet none of them focused on its excretory/secretory (E/S) proteins that easily get missed when working solely on extracts from crushed nematodes. Moreover, these E/S allergens remain behind even when the larva has been removed during fish quality processing. Therefore, the aim was to investigate if Anisakis-like allergens could also be detected in both crushed and E/S P. decipiens protein extract using targeted mass spectrometry analysis and immunological methods. The results confirmed that at least five A. simplex allergens have homologous proteins in P. decipiens; a result that emphasizes the importance of also including E/S protein extracts in proteomic studies. Not only A. simplex, but also P. decipiens should therefore be considered a potential source of allergens that could lead to hypersensitivity reactions in humans.
Topics: Allergens; Animals; Anisakis; Ascaridoidea; Fishes; Hypersensitivity; Immunoassay; Larva; Phylogeny; Proteomics
PubMed: 35260766
DOI: 10.1038/s41598-022-08113-3 -
Turkiye Parazitolojii Dergisi Mar 2024is a fish-borne zoonotic parasite. Although this parasite has been known since the 19 century, publications on anisakiasis have increased in recent years. Despite this,...
OBJECTIVE
is a fish-borne zoonotic parasite. Although this parasite has been known since the 19 century, publications on anisakiasis have increased in recent years. Despite this, this subject has yet to be well studied by bibliometric analysis. This study was conducted to show the research points and trends of .
METHODS
The Web of Science Core Collection (WoSCC) was mined for articles on . The VOSviewer software visually evaluated countries, institutions, authors, references, and keywords in this field.
RESULTS
A total of 1362 publications were included in this bibliometric analysis. The included publications were published between 1970 and 2022 from 79 countries, mainly from Spain (n=456, 33.48%). The most prolific year was 2020 (n=74). The research area that attracted the most publications was parasitology (n=452), while the most productive author in this area was Cuellar C (n=53). , and were the most used three keywords.
CONCLUSION
The number of publications on anisakiasis has been increasing over time, suggesting that is becoming an increasingly important disease worldwide. Research cooperation should be established between researchers from developed and developing countries to determine effective control strategies for anisakiasis.
Topics: Animals; Anisakis; Anisakiasis; Bibliometrics; Software
PubMed: 38449368
DOI: 10.4274/tpd.galenos.2024.94830 -
International Journal For Parasitology.... Aug 2024We here describe the location of anisakid third stage larvae in Atlantic herring L. caught in the North Sea in August 2023. We further demonstrate how industrial...
We here describe the location of anisakid third stage larvae in Atlantic herring L. caught in the North Sea in August 2023. We further demonstrate how industrial processing (mechanical gutting, removal of entrails, head, tail, hypaxial anterior musculature and vertebral column) reduces the overall infection and worm load in the musculature. The isolated anisakid larvae were identified as sensu stricto by a combination of morphometrics and molecular methods (PCR of rDNA and mtDNA, sequencing, BLAST analysis). As a baseline we examined a total of 75 specimens of freshly caught and ungutted herring and showed a positive correlation between host size (fish length and weight) and infection level. The overall prevalence of infection was 84 %, the mean intensity 11.3 (range 1-38 parasites per fish) and the abundance 9.52. The main part of the overall worm population was associated with stomach and pyloric caeca in the body cavity (77 %) and only 5 % was found in the musculature. Larvae occurred in the hypaxial part of the musculature (21), the epaxial part (7 worms) and the caudal part (5 worms). The prevalence of muscle infection was 28 % and the mean intensity 1.6 (range 1-5) parasites per fish and abundance 0.44 parasites per fish. In order to assess the effect of industrial processing on worm occurrence in the fish we examined a total of 67 specimens of herring, from exactly the same batch, but following processing. This included removal of organs in the body cavity, cutting the lower part of the hypaxial segment but leaving the right and left musculature connected by dorsal connective tissue. Five out of these fish carried one larva (prevalence 7.5%, mean intensity 1, abundance 0.07 larvae per fish), and these worms were located in the ventral part of the anterior musculature (2), in the central part of the anterior musculature (2) and one larva in the central part of the caudal musculature. The industrial processing reduced the overall occurrence (abundance) of worms in the fish from 9.52 to 0.07 (136 times reduction) and the occurrence in the musculature from 0.44 to 0.07 (6.28 times reduction). The overall prevalence was reduced from 84 % to 7.5 % (11.2 times reduction). Muscle infection prevalence fell from 28 % to 7.5 % (3.7 times reduction). We then followed another batch of herring following a marinating process (11% NaCl for 24 h and subsequent incubation in acetic acid and vinegar) by artificially digesting the flaps during week 1-8. Although a total of 31 larvae were recovered from 144 fish examined no live nematode larvae were isolated. The importance of fish handling, processing and marination for consumer safety is discussed.
PubMed: 38655447
DOI: 10.1016/j.ijppaw.2024.100937 -
Parasitology Research Mar 2018Anisakiasis 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...
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 LD 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.
Topics: Animals; Anisakiasis; Anisakis; Anthelmintics; Cholinesterase Inhibitors; Cymenes; Fish Diseases; Fishes; Gas Chromatography-Mass Spectrometry; Larva; Monoterpenes; Oils, Volatile; Origanum; Plant Oils; Spain; Thymol
PubMed: 29368038
DOI: 10.1007/s00436-018-5764-3 -
Anales Del Sistema Sanitario de Navarra 2003Anisakis simplex is a parasite, belonging to the Anisakidae family. The life cycle of the parasite can include one or more intermediary hosts, their final hosts being... (Review)
Review
Anisakis simplex is a parasite, belonging to the Anisakidae family. The life cycle of the parasite can include one or more intermediary hosts, their final hosts being marine mammals or large fish, in which the larvae develop until the adult stage is reached. Man is an accidental host who acquires the larvae by eating raw or undercooked fish. Since the mid-50s, when the first case studies were published in Holland and Japan, new cases have been emerging in different countries including Spain. Parasitization of man by the living larva is known as anisakiasis, principally giving rise to digestive symptomology, with other rare cases described of invasion of other organs such as the lung, the liver, the spleen, the pancreas, etc. Clinical pictures of allergy to IgE mediated anisakis simplex have also been described: reactions by thermostable antigens of the parasite that develop in spite of the fish being cooked or frozen, and an acute digestive parasitization with allergic symptoms called gastro-allergic anisakiasis. In the diagnosis of anisakiasis and/or allergy to Anisakis, the antecedent of the prior ingestion of fish as well as the clinical accompaniment can form basic data of considerable orientational value, and endoscopy can reveal the presence of the larvae and make possible their extraction. Besides, in cases of allergy the detection test for specific IgE facing Anisakis simplex, and cutaneous tests with fish should be carried out. The best treatment for avoiding this parasitization is prophylactic, avoiding the consumption of raw or undercooked fish, while a fish free diet is necessary in cases of true allergy to the thermostable proteins of the parasite.
Topics: Anisakiasis; Humans; Hypersensitivity
PubMed: 13679960
DOI: No ID Found