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Anisakis are foodborne parasites that opportunistically parasitize humans, leading to acute abdominal symptoms and allergies. Besides gastroscopy, no other diagnostic technique is available. Consequently, it is necessary to identify specific biomarkers and then develop molecular techniques for diagnosing Anisakis infection. In the present study, we used immunoproteomic and immunoinformatic approaches to identify sensitive antigens for diagnosing Anisakis pegreffii infection. A total of three proteins, including Ani609 (VDK51609), Ani941 (VDK75941), and AniS13, were identified based on immunoinformatic results. Then, the indirect ELISA method was developed based on the recombinant proteins, showing a similar diagnostic capability to that of parasitic soluble proteins. Next, a Gaussia luciferase immunoprecipitation assay (LIPS) was further developed upon the fusion of the proteins and Gaussia luciferase. The LIPS method indicated that A. pegreffii infection could be detected in rats as early as 1 week post infection, especially for Ani941. Overall, we identified the novel antigens using immunoproteomic and immunoinformatic approaches and then developed a sensitive method for diagnosing A. pegreffii infection, particularly for the early stage.

Patient aged 71 with a history of type 2 diabetes mellitus. He came to the emergency department for abdominal pain and vomiting. Laboratory tests showed an increase in acute phase reactants. Abdominal CT scan showed dilated jejunal loops, compatible with intestinal occlusion. Urgent intervention was performed, resecting the affected segment. The pathology report showed a prominent transmural inflammatory infiltrate and interstitial oedema, with moderate villous atrophy, identifying parasitic structures compatible with anisakis larvae (family Anisakidae). Given the mechanism of tissue invasion, the larvae are surrounded by a predominantly eosinophilic inflammatory infiltrate, organised as granulomas or abscesses.

Historical seroprevalence data for Anisakis in Spain vary greatly depending on the sampling region owing to different fish consumption habits. As a result of European Regulation (EC) No. 853/2004, the Royal Decree 1420/2006 on the prevention of parasitosis by Anisakis in fishery products supplied by establishments that serve food to final consumers or to communities came into force in Spain. In this study, a prevalence study of Anisakis in Madrid has been conducted to verify the prophylactic effects of the application of the law. Sera from 500 blood donors from the Fundación Jiménez Díaz University Hospital (Madrid/2021-2023) were collected, and the levels of anti- Anisakis IgG, IgA, and IgE were analyzed by ELISA, comparing them with those obtained with 110 donors from the Red Cross and the "Gómez Ulla" Central Defense Hospital (Madrid/2001-2002). The percentages of positivity in the 2021-2023 donor group were IgG (13.6%), IgA (13.6%), and IgE (2.2%), while in the 2001-2002 donors they were positive for IgG (15.45%), IgA (14.54%), and IgE (11.65%). A reduction of more than 80% was observed in the prevalence of anti- Anisakis IgE in the healthy population of Madrid, which confirmed the positive effect of RD1420/2006, which was later incorporated into RD1021/2022.

Background: Anisakiasis, a zoonotic disease caused by the nematode Anisakis, poses a significant concern for public health, particularly in regions with high consumption of raw or undercooked fish., Case Presentation: We present a case report of a 41-year-old woman who developed severe abdominal symptoms, ultimately diagnosed with intestinal obstruction due to Anisakis infestation, requiring surgery. Despite the absence of prominent eosinophilia or specific radiological findings, the diagnosis was confirmed through histological examination, highlighting the importance of considering anisakiasis in patients with a history of raw seafood consumption., Conclusion: The case underscores the diagnostic challenges associated with anisakiasis, emphasizing the need for increased awareness among healthcare professionals and the public regarding the risks of consuming raw or undercooked seafood. Effective management requires a multidisciplinary approach, including clinical assessment, imaging studies, and histological evaluation, to ensure timely diagnosis and appropriate treatment., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Anisakiasis is a parasitic disease transmitted through the consumption of raw or undercooked fish and cephalopods that are infected with larvae of Anisakis simplex (sensu stricto) or Anisakis pegreffii. The purpose of this study was to investigate how A. simplex (s. s.) responds to the influence of anthelmintics such as ivermectin (IVM) and pyrantel (PYR). In vitro experiments were conducted using larvae at two developmental stages of A. simplex (s. s.) (L3 and L4) obtained from Baltic herring (Clupea harengus membras). Larvae were cultured with different concentrations of IVM or PYR (1.56, 3.125, and 6.25 μg/mL) for various durations (3, 6, 9, and 12 h) under anaerobic conditions (37 °C, 5% CO 2 ). The gene expression of actin, ABC transporter, antioxidant enzymes, γ-aminobutyric acid receptors, and nicotinic acetylcholine receptors, as well as the oxidative status were analyzed. The results showed that A. simplex (s. s.) L3 stage had lower mobility when cultured with PYR compared to IVM. The analysis of relative gene expression revealed significant differences in the mRNA level of ABC transporters after treatment with IVM and PYR, compared to the control group. Similar patterns were observed in the gene expression of antioxidant enzymes in response to both drugs. Furthermore, the total antioxidant capacity (TAC) and glutathione S-transferase (GST) activity were higher in the treatment groups than in the control group. These findings suggest a relationship between the expression of the studied genes, including those related to oxidative metabolism, and the effectiveness of the tested drugs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Anisakis can cause Anisakiasis in humans if raw or undercooked fish is consumed. Symptoms of infection may include vomiting, acute abdominal symptoms, or allergies. In this study, we collected 187 commercially available marine fish from the Yellow Sea, East China Sea, and South China Sea. Among them, 79 were found positive containing 520 Anisakis worms. The average prevalence rate was found 42% in this investigation. Ninety-two worms from different sea areas were selected and analyzed for identification, revealing the presence of five different species, which are Anisakis pegreffii , Hysterothylacium aduncum , Hysterothylacium zhoushanense , Hysterothylacium amoyense , and Hysterothylacium sp. In the meta-analysis, three databases: PubMed, CNKI, and BaiduXueshu were searched for surveys on the prevalence of Anisakis in Chinese waters from January 2000 to December 2023. A total of 26 studies were included in this analysis of which 25 publications were retrieved from different databases and one being the present study. The pooled prevalence of Anisakis was 45% among commercially available marine fish. Variances in the prevalence of Anisakis were noted among the four seas, with the highest rates in the East China Sea and the Bohai Sea, reaching 53% [0.38; 0.68] and 49% [0.36; 0.62], respectively. The Prevalence of Anisakis infection was significantly higher in astern parts such as Liaoning, Shanghai, and Zhejiang. Analysis of the host fish subgroups revealed that the orders of Anguilliformes, Scombriformes, and Gadiformes had high rates of infection. These findings suggest a significant prevalence of Anisakis , posing an increasing risk of infection for individuals. This study provides impactful information for implementing preventative measures against Anisakis ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zeng, Fang, Wang, Qadeer, Qiu, Hong and Mahmoud.)

Many pathogens are related to carcinogenesis. Chronic inflammation, as a result of persistent infection, leads to DNA damage, higher expression of oncogenes, decreased apoptosis and immunosuppression, which are some of the reasons for cancer induction. Among parasites, Schistosoma, Opistorchis and Clonorchis are recognised as infectious agents which contribute to cancer. A relationship between Anisakis and cancer was hypothesised because cellular responses to Anisakis products could result in inflammation and DNA damage. Previous research has shown a decrease in CD8+ γδ T-cells and an increase in αβ and γδ T-cell apoptosis in colon cancer (CC) samples. Ninety-two CC patients and 60 healthy subjects were recruited. γδ and αβ T-cells were analysed, and their apoptosis was evaluated. Anti-Anisakis antibodies were tested in sera from CC patients and controls. Anti-Anisakis IgG, IgM, IgA and IgE antibodies were significantly higher in CC patients. A significant increase in anti-Anisakis IgA levels was observed in patients with angiolymphatic invasion. The number of all γδ T-cells, as well as CD3+ CD4+ αβ T-cells, was significantly lower in CC patients. The apoptosis of all T-cells was significantly increased in patients with CC. We observed a significantly higher percentage of anti-Anisakis IgE positive patients having a deficit of CD3+ γδ T-cells. Our results suggest a relationship between Anisakis and CC., (© 2024. The Author(s).)

We here describe the location of anisakid third stage larvae in Atlantic herring Clupea harengus 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 Anisakis simplex 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., Competing Interests: The authors all declare that they have no conflict of interests., (© 2024 The Author(s).)

In the last decade, it has become evident that various RNA viruses infect helminths including Order Ascaridida. However, there is still no information available for viruses infecting Anisakis. We herewith demonstrate the presence of a novel rhabdovirus from Anisakis larvae detected by next-generation sequencing analysis and following RT-PCR. We determined the nearly all nucleotide sequence (12,376 nucleotides) of the viral genome composed of seven open reading frames, and we designated the virus as Suzukana rhabdo-like virus (SkRV). BLASTx search indicated that SkRV is a novel virus belonging to the subfamily Betanemrhavirus, rhabdovirus infecting parasitic nematodes of the Order Ascaridida. SkRV sequence was detectable only in the total RNA but not in the genomic DNA of Anisakis, ruling out the possibility of SkRV being an endogenous viral element incorporated into the host genomic DNA. When we individually tested Anisakis larvae obtained from Scomber japonicus migrating in the coastal waters of Japan, not all but around 40% were SkRV-positive. In the phylogenetic trees of Betanemrhavirus and of the host Ascaridida nematodes, we observed that evolutional distances of viruses were, to some extent, parallel with that of host nematodes, suggesting that viral evolution could have been correlated with evolution of the host. Although biological significance of SkRV on Anisakis larvae is still remained unknown, it is interesting if SkRV were somehow related to the pathogenesis of anisakiasis, because it is important matter of public health in Japan and European countries consuming raw marine fishes., Competing Interests: Declaration of Competing Interest There is nothing to be declared of COI condition in our study., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: To investigate the prevalence of Anisakis infections in coastal marine fishes and awareness of anisakiasis control knowledge among local residents in Yantai City, Shandong Province in 2021, so as to provide insights into formulation of anisakiasis control interventions., Methods: Marine fishes were purchased from Shunxin Port, Yantai City, Shandong Province in November 2021, and the presence of Anisakis was detected in different species of fishes and different fish sites. The correlations between body length and weight of marine fish and intensity of Anisakis infections were examined using Spearman's rank correlation analysis, and the dietary habits and anisakiasis control knowledge were investigated using questionnaire surveys among local residents., Results: A total of 201 marine fishes belonging to 20 species were dissected, and Anisakis was detected in 77 marine fishes (38.31%) belonging to 11 species (55.00%), with a mean infection intensity of 45.04 parasites per fish (3 468/77). Spearman's rank correlation analysis revealed that the body length ( r s = 0.74, P < 0.05) and weight ( r s = 0.79, P < 0.01) of the monkfish correlated positively with the intensity of Anisakis infections, and the body length ( r s = 0.68, P < 0.05) of the flatfish correlated positively with the intensity of Anisakis infections, while no correlations were examined between the body length or weight of other marine fishes and the intensity of Anisakis infections. Of all respondents, 53.38% men and 56.67% women did not know anisakiasis control knowledge at all, and there was a significant difference in the proportion of respondents using separate chopping boards for raw and cooked food from different villages (χ 2 = 17.89, P < 0.01), while there was an age-specific proportion of respondents with habitats of eating raw or semi-raw seafood (χ 2 = 28.27, P < 0.01)., Conclusions: The prevalence and intensity of Anisakis infections were high in coastal marine fishes in Yantai City in 2021, and the awareness of anisakiasis control knowledge was low among local residents. Intensified health education pertaining to anisakiasis control knowledge is recommended to reduce the risk of Anisakis infections.

The phenotype of allergic diseases associated with Anisakis determines the pattern of cytokines related to antibody production. However, the role of serum IgA and the immunomodulatory mechanisms exerted by active infection of L3 or passive mucosal contact with A. simplex specific antigens has not been studied before. We measured serum cytokine by flow cytometry (IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ, IL-17A, TGF-β1) and antibody levels (IgE, IgG4, IgA) by ELISA against total and excretory-secretory (ES) antigens, Ani s 3,and the group of major allergens Ani s 1, Ani s 7, and Ani s 13 in sera from 10 patients with gastro-allergic anisakiasis (GAA), 11 Anisakis sensitization associated chronic urticaria (CU+) as well as 17 non-Anisakis-sensitized patients with chronic urticaria (CU-), compared with the urticaria control group (18 subjects). Specific IgE, IgG4 and IgA were high in the GAA, but IgA levels were significantly higher in the CU+ with respect the CONTROL group. We observed higher levels of the ratio IgA/IgG4 in CU+ than GAA group for Ani s 1, Ani s 7, Ani s 13 and ES. Furthermore, chronic urticaria (CU) patients showed significant lower levels of IL-10, IFN-γ and IL-17A than patients without CU. The anti-Ani s 13 IgA/IgG4 ratio correlated positively with pro-inflammatory cytokines and ratios (TNF-α, IL-17A, Th17/Th2, Type1/Type2 and TNF-α/IL-10) in CONTROL group. In general, Anti-Anisakis IgA/G4 ratio was high in CU patients. In conclusion, this study demonstrates the importance of serum IgA because it is associated with chronic urticaria independently of Anisakis sensitization., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Objective: Anisakis simplex is a fish-borne zoonotic parasite. Although this parasite has been known since the 19 th 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 A. simplex ., Methods: The Web of Science Core Collection (WoSCC) was mined for articles on A. simplex . 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). "Anisakis simplex" , "Anisakis" and "anisakiasis" were the most used three keywords., Conclusion: The number of publications on anisakiasis has been increasing over time, suggesting that A. simplex 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.

Anisakiasis is a parasitic infection caused by the ingestion of raw or undercooked seafood infected with Anisakis larvae. It generally affects the gastrointestinal tract, particularly the stomach, but very rare cases have been reported in which infection of the liver leads to the formation of inflammatory pseudotumors. We herein report an extremely rare case of an inflammatory pseudotumor induced by hepatic anisakiasis that was laparoscopically resected for the purpose of both diagnosis and treatment. A 51-year-old woman underwent a routine medical checkup by ultrasound examination, which incidentally detected a 15-mm mass on the surface of S6 of the liver. Because a malignant tumor could not be ruled out on several preoperative imaging studies, laparoscopic partial resection of the liver was performed. Histopathological examination revealed Anisakis larva in the inflammatory pseudotumor, suggesting hepatic anisakiasis. This report describes an extremely rare case of an inflammatory pseudotumor induced by hepatic anisakiasis. Because the preoperative diagnosis could not be obtained by several imaging modalities, laparoscopic liver resection with a sufficient margin might be suitable for diagnosis and treatment of this disease., (© 2023. Japanese Society of Gastroenterology.)

Zoonotic nematodes of the family Anisakidae are highly common in many marine fish species, which act as paratenic hosts for the third larval stage. In the fish, these parasites may migrate from the fish's gastro-intestinal tract (GI-tract) further to the coelomic cavity and muscles, making them a possible contamination source of bacteria they carry on their cuticle and in their GI-tract. A previous study revealed no apparent effect of Anisakis simplex on spoilage of fish, but the equally common anisakid species Pseudoterranova decipiens has a larger body surface potentially increasing the bacterial load brought into the fish muscle upon migration. As the presence of shelf-life reducing spoilage bacteria in the microbiome of this anisakid species has been demonstrated, the objective of the present study was to assess the potential shelf-life reducing effect of P. decipiens in fresh fish fillets stored in a domestic refrigerator. Atlantic cod was used as a model since members of the cod family are the third most consumed marine fish globally and it has the highest prevalence of P. decipiens infections. Infected and non-infected codfish fillet portions were collected and microbiologically analyzed at day 0 and day 4 of storage in a domestic fridge. Three isolation media were used to enhance maximum bacterial recovery and isolates were identified using MALDI-TOF MS and 16S rRNA gene sequencing. In parallel to the microbiological examination, sensory analysis was performed daily on the cod fillets to evaluate the freshness of the fish. Results revealed the presence of typical spoilage bacteria (e.g., Pseudomonas sp., Photobacterium sp.) in all fish, but based on the total viable counts, total H 2 S-producing bacteria, and sensory analysis, there were no objective indications to assume an increased fish spoilage rate by the presence and migration P. decipiens. Additionally, a beta-diversity comparison revealed no significant differences in microbiota composition between infected and non-infected fish parts, though individual heterogeneity in microbiome composition among Atlantic codfish individuals was found. As total viable counts did, however, exceed the guideline limits for fresh fish, further research should now focus on the role of the candling step as a potential source of post-harvest contamination. As such, anisakid infection might still accelerate fish spoilage, though now in an indirect way., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

The European anchovy Engraulis encrasicolus is one of the most important commercial species in the Bay of Biscay (ICES Subarea 8), and our analysis focused on the analysis of the temporal mean abundance, prevalence, and intensity of Anisakis spp. larvae species in anchovies from ICES Subarea 8 in the years 2000, 2001, 2014-2016, and 2019-2023. Prevalence in adult individuals of anchovy was only 1% in 2000 but increased to 90% in 2014. Since 2015, the prevalence has decreased, and the number of individuals affected in 2023 accounted for 17.6%. The mean abundance showed a similar trend, with a peak of 3.79 nematodes/anchovy in 2014, falling to 0.21 in 2023. The species A. simplex sensu stricto and A. pegreffii were identified by PCR/SANGER sequencing and PCR/RLFP techniques in 2019 and 2020. Anisakis simplex (s.s.) was the most abundant species and, according to the results returned by these two techniques, it accounted for an average of 62.4% and 52.1% of total nematodes in 2019 and 2020, respectively. The results of studies monitoring infection levels in anchovies showed that the mean abundance and prevalence changed over the course of the study period and that the proportion of different species of Anisakis is also subject to variation from year to year., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Globally, the exploitation of small pelagic fish, like Pacific mackerel is of great importance due to food industry demand. However, there are few studies regarding its parasites load and there are no in this geographic zone. This study aimed to assess the parasitic composition, some temporal changes (during spring and summer) in abundance, prevalence and intensity of infection parasitic of the Pacific mackerel ( Scomber japonicus ) from Todos Santos Bay, Baja California, Mexico. The parasite fauna of the Pacific mackerel consisted of 1930 parasites (1413 in spring and 517 in summer) distributed in the follow taxa: an Tetraphyllidea (Cestoda), Kuhnia scombri (Monogenea), Didymozoidae (Digenea), Anisakis sp. (Nematoda), Rhadinorhynchus sp. (Acanthocephala) and Caligus pelamydis (Copepoda). The nematodes parasite were the most abundant both in spring with a mean abundance of 27.6 parasites and in summer 8.2 parasites compared with the other taxa like Cestoda, Monogenea, Digenea, Acanthocephala and Copepoda ( P = 0.003). The mean intensity of the nematodes in spring and summer was 28.1 and 13.4, respectively. The nematodes prevalence was 90 % in spring and 60 % in summer. In general, the parasite load is more abundant in spring than summer. In summer, absence of taxa as Cestoda and Copepoda were registered. Nematode larvae were present in the fish guts mesentery and inside of the stomach, pyloric caeca, intestine. Also the nematodes were found in the liver, muscle and gonads. The most affected organ by nematodes was the intestine mesentery. The most predominant parasite of this study has been Anisakis sp. during spring., Competing Interests: Conflict of Interest Authors state no conflict of interest., (© 2023 M. Vivanco-Aranda et al., published by Sciendo.)

Fish play an important role in human nutrition. They are not only a great source of protein and healthy fats, but also a unique source of essential nutrients such as omega-3 fatty acids. Moreover, most fish are parasitized, and some of these parasites are able to influence the reallocation of resources in their favor and thus reduce the nutritional quality of the fish. The present study was conducted to investigate the impact of the third stage larvae (L3) of Anisakis spp. on the proximate composition, macro-minerals (potassium, calcium, and sodium), and fatty acids of European hake (Merluccius merluccius Linnaeus, 1758). In parasitized female group, our results revealed a decrease (p< 0.005) in the amount of carbohydrate by 6.5%, of calcium by 17%, and of 2 polyunsaturated fatty acids (arachidonic acid (C20: 4w-6), and eicosapentaenoic acid (C20: 5w-3) with 33% and 15% respectively. Simultaneously, an increase by 25% in the level of a single saturated fatty acid C10:0 was noticed. According to the principal component analysis, the parasitized female was wealthy of saturated fatty acids and monounsaturated fatty acids and contains less of polyunsaturated fatty acids, omega-3 fatty acids, and omega-6 fatty acids than the unparasitized female and male. No significant changes were observed in the biochemical composition of male hake, probably due to the low mean intensity of L3 larvae of Anisakis spp. in this group., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Fish products in Slovakia have been heavily infected with Anisakis spp. larvae, which causes human anisakiasis. We found larvae in all tested samples of frozen Atlantic herring. Anisakid allergen t-Ani s7 testing revealed 2 positive cases in humans, signaling need for health authorities to closely monitor zoonotic marine parasites, even in inland areas.

Anisakiosis is a significant zoonotic disease caused by parasitic nematodes of the Anisakis genus. It can be contracted by humans through the consumption of raw or undercooked fish contaminated with the parasite, leading to gastrointestinal and allergic symptoms. While anisakiosis is not frequently documented in Portugal, the presence of allergic reactions to Anisakis in Spain suggests ongoing exposure in the Iberian Peninsula. To address this concern, the Interdisciplinary Centre of Marine and Environmental Research in Porto, Portugal, in collaboration with the Biology Centre of Czech Academy of Sciences in Ceske Budejovice, Czech Republic, has proposed a project entitled 'Assessing Portuguese Health Risks: Anisakis Parasite in Atlantic Chub Mackerel ( Scomber colias ) Sold in Portuguese Markets' under the European Food Risk Assessment Fellowship Programme. The primary objective of the project is to gather valuable epidemiological data on the host, Atlantic chub mackerel ( S. colias ) and the parasitic nematode ( Anisakis spp.) with the focus on assessing contamination levels and evaluating potential health risks associated with anisakiosis in the Portuguese population. By conducting this research, the project aims to contribute to the understanding of anisakiosis and its impact on public health in Portugal. Investigation of the presence of the Anisakis parasite in Atlantic chub mackerel sold in Portuguese markets will provide crucial insights into the risks associated with consuming raw or undercooked fish. Ultimately, our findings will aid in the development of preventive measures and guidelines to ensure the well-being of the Portuguese population., (© 2023 European Food Safety Authority. EFSA Journal published by Wiley‐VCH GmbH on behalf of European Food Safety Authority.)

The European conger, Conger conger , is a benthic marine fish species with a geographical distribution extending through the northeastern Atlantic and the Mediterranean. Despite being extensively distributed and widely appreciated by Spanish consumers, studies regarding parasite presence in this fish are scarce. In the present work, a hundred and eight specimens from the Mediterranean coast of northeastern Spain (Catalan waters) were surveyed for the presence of nematode parasites. Several species were morphologically identified: third-stage larvae of Anisakis type I (sensu Berland, 1961) ( n = 131), third-stage larvae of Anisakis type II (sensu Berland, 1961) ( n = 18), third- and fourth-stage larvae and adults of Hysterothylacium spp. ( n = 48), adults of Cucullanus sp. ( n = 391), and adults of Cristitectus congeri ( n = 69). Moreover, some fish and decapode species were also observed as part of the host's diet, with the most detected preys being Micromesistius poutassou , Sardina pilchardus , Macropipus sp., and Goneplax rhomboides . This represents the first survey of nematode parasites infecting C. conger from the northeastern Spanish Mediterranean waters. Among the parasite species detected, the presence of Anisakis species should be highlighted as the ingestion of C. conger parasitized with these larvae could potentially lead to anisakiasis in consumers.

The majority of research on the safety of marine edible fish has primarily focused on anisakid nematodes, neglecting the potential risks posed by other parasites, including those belonging to the family Gnathostomatidae. In Australia, there have been few reported cases of human infections with gnathostomatid parasites since 2011. However, due to the absence of a standardized diagnostic test in the country, it is believed that the actual number of infections is higher than reported. This study aimed to assess the occurrence and prevalence of infectious gnathostomatid parasites in selected commercial fish species in Australia. A total of 1947 marine fish from northern Australia, representing 9 families, 16 genera, and 30 species, were examined for gnathostomatid nematode infections. Overall, 12.3 % of the fish were found to be infected with at least one gnathostomatid larva. Among the species examined, the yellow-dabbled flounder (Branchypleura novaezeelandiae) exhibited the highest prevalence (83.3 %; n = 6) and the largest number of gnathostomatid larvae. The identification of the gnathostomatid larvae was confirmed as belonging to the genus Echinocephalus based on both morphological characteristics and sequence data. No significant correlation was observed between the prevalence, mean abundance, and mean intensity of infection with the length or weight of the examined fish species. Notably, several of the infected fish species are considered popular choices in the Australian market. Hence, it is imperative to raise awareness among relevant food safety authorities regarding the occurrence of these parasites. The findings from this study should be taken into consideration for the revision of current seafood safety protocols in the country., Competing Interests: Declaration of competing interest All authors declare there is no conflict of interest to report., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

This survey focuses on an environmental parasitology study by investigating Polycyclic Aromatic Hydrocarbons (PAH) bioaccumulation and the occurrence of Anisakis spp. on common anchovies collected from Moroccan coasts over 9 months through chemical (PAH) and Biological (Biometrics and parasitic) analysis. Obtained results were statistically analyzed and human health risks from anchovies consumption were assessed. The results obtained highlighted the good biological and chemical status of this fish in all stations of Morocco's coasts. Anisakis spp. was present in only four stations in the Atlantic fringe (maximum prevalence 22.22 %). Results have shown non-significant differences for tissue nature (Muscle, viscera) or spatial variation and were within a low range of concentrations well below the European Commission standards. Results have shown no serious harmful risk for humans from this fish consumption (ILCR and HI), and statistical analysis had shown positive correlations between prevalence and Chr, prevalence and sex-ratio, and prevalence and weight., Competing Interests: Declaration of competing interest We have no conflicts of interest to disclose. All authors declare that they have no conflicts of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

A review was conducted to identify the most common causative agents of anisakidosis, the methods used for identification of the causative agents, and to summarize the sources of infection, and patients' demographics. A total of 762 cases (409 articles, inclusive of all languages) were found between 1965 and 2022. The age range was 7 months to 85 years old. Out of the 34 countries, Japan, Spain, and South Korea stood out with the highest number of published human cases of anisakidosis, respectively. This raises the question: Why are there few to no reports of anisakidosis cases in other countries, such as Indonesia and Vietnam, where seafood consumption is notably high? Other than the gastrointestinal tract, parasites were frequently found in internal organs such as liver, spleen, pancreas, lung, hiatal and epigastric hernia, and tonsils. There are also reports of the worm being excreted through the nose, rectum, and mouth. Symptoms included sore throat, tumor, bleeding, gastric/epigastric/abdominal/substernal/lower back/testicular pain, nausea, anorexia, vomiting, diarrhea, constipation, intestinal obstruction, intussusception, blood in feces, hematochezia, anemia, and respiratory arrest. These appeared either immediately or up to 2 months after consuming raw/undercooked seafood and lasting up to 10 years. Anisakidosis commonly mimicked symptoms of cancer, pancreatitis, type I/II Kounis syndrome, intussusception, Crohn's disease, ovarian cysts, intestinal endometriosis, epigastralgia, gastritis, gastroesophageal reflux disease, hernia, intestinal obstruction, peritonitis, and appendicitis. In these cases, it was only after surgery that it was found these symptoms/conditions were caused by anisakids. A range of not only mainly marine but also freshwater fish/shellfish were reported as source of infection. There were several reports of infection with >1 nematode (up to >200), more than one species of anisakids in the same patient, and the presence of L4/adult nematodes. The severity of symptoms did not relate to the number of parasites. The number of anisakidosis cases is grossly underestimated globally. Using erroneous taxonomic terms, assumptions, and identifying the parasite as Anisakis (based solely on the Y-shaped lateral cord in crossed section of the parasite) are still common. The Y-shaped lateral cord is not unique to Anisakis spp. Acquiring a history of ingesting raw/undercooked fish/seafood can be a clue to the diagnosis of the condition. This review emphasizes the following key points: insufficient awareness of fish parasites among medical professionals, seafood handlers, and policy makers; limited availability of effective diagnostic methodologies; and inadequate clinical information for optimizing the management of anisakidosis in numerous regions worldwide., (© 2023. The Author(s).)

Background: Gastric anisakiasis typically causes severe abdominal symptoms; however, we incidentally detected asymptomatic gastric anisakiasis cases during esophagogastroduodenoscopy. The factors associated with developing acute abdominal symptoms induced by gastric anisakiasis remain unclear. Therefore, this study aimed to investigate the clinical factors associated with abdominal symptoms of gastric anisakiasis by comparing symptomatic and asymptomatic cases., Methods: This was a retrospective cohort study involving 264 patients diagnosed with gastric anisakiasis at nine hospitals in Japan between October 2015 and October 2021. We analyzed patients' medical records and endoscopic images and compared the clinical factors between the symptomatic and asymptomatic groups., Results: One hundred sixty-five patients (77.8%) were diagnosed with abdominal symptoms, whereas 47 (22.2%) were asymptomatic. Older age, male sex, diabetes mellitus, gastric mucosal atrophy, and gastric mucosal atrophy of the Anisakis penetrating area were significantly more common in the asymptomatic group than in the symptomatic group. Multivariate analysis revealed that age (p = 0.007), sex (p = 0.017), and presence or absence of mucosal atrophy (p = 0.033) were independent factors for the occurrence of acute abdominal symptoms. In addition, cases that were Helicobacter pylori naïve, with an elevation of white blood cells, or without an elevation of eosinophils were more common in the symptomatic group than in the asymptomatic group., Conclusions: Age, sex, and presence or absence of gastric mucosal atrophy were the clinical factors associated with the occurrence of acute abdominal symptoms. Older and male patients and those with gastric mucosal atrophy were less likely to show abdominal symptoms. The mechanisms of the occurrence of symptoms induced by gastric anisakiasis remain unclear; however, our results will help clarify this issue in the future., (© 2023. The Author(s).)

Anisakiasis is an arising zoonosis induced by parasitic nematodes belonging to the family Anisakidae. Anisakiasis is often caused by the ingestion of larval nematodes in uncooked or minimally processed seafood dishes, which are regularly consumed by humans. Significant potential sources of infection are raw fish (e.g., sushi and sashimi) that can be found in traditional Japanese cuisine and can be part of the culinary tradition of consumption of raw or marinated fish that is particularly diffused in European countries. During the last five decades, the global prevalence of human anisakiasis has been rising, becoming an emergent major public health problem. Thus, there is an unmet need for well-defined and cost-effective methods aimed at killing Anisakis larvae, thus reducing the incidence of anisakiasis. In this mini-review, we discuss the clinical features of anisakiasis as well as the effectiveness and mechanisms of action of the main methods employed for increasing seafood safety and killing Anisakis larvae, including freezing, heating, use of high hydrostatic pressure, salting process, pepsin digestion method and use of garlic oil.

Human anisakiasis is a foodborne disease that has been increasingly reported worldwide and is caused by the consumption of raw or undercooked seafood infected with zoonotic nematodes of the genus Anisakis Dujardin, 1845. One of the most frequently reported species, both in fish paratenic hosts as well as in human patients, is Anisakis simplex sensu stricto (s.s.), which is distributed within some of the globe's main fishing grounds in the North Pacific and Atlantic oceans. In order to clarify the influence of temperature on this parasite in vivo, third-stage larvae of A. simplex s.s. were surgically challenged in the body cavities of rainbow trouts (Oncorhynchus mykiss) and Mozambique tilapias (Oreochromis mossambicus). Larval survival and tissue migration were then analyzed after 6 and 12 weeks. The results showed that survival rates of larvae were lower at 27 and 33 °C than at 3, 9, 15 and 21 °C. Also, migration to the body muscle was observed to be highest at 9 °C. These results suggest that third-stage larvae of A. simplex s.s. are more adapted to lower temperatures in experimentally challenged fish, which may justify its distribution in cold northern waters., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The Anisakis larvae presence in fish for human consumption is a health risk that needs to be monitored. The anchovy is a fish that is highly appreciated by consumers and that can harbour Anisakis. It is thus necessary to periodically evaluate the presence of anisakid larvae in them. So, anchovies from Iberian Peninsula coasts were analysed. Fish examination for macroscopic nematodes showed L3s of both Anisakis type I and Hysterothylacium aduncum. The Anisakis prevalence varies with the catching area and the fish size. The muscle prevalence was 7.45% (mean intensity 1.75; range 1-5). Molecular analysis showed 110 A. simplex s.s. (17 in muscle), 22 A. pegreffii (3) and 7 hybrid genotype individuals (1). Considering that most of the Iberian Peninsula coasts are a sympatry area between these two Anisakis species, it has been observed that A. simplex s.s./A. pegreffii ratio increases from south to north in a clockwise direction. Also, 19 larvae were detected on the fish surface from the Bay of Biscay, indicating the ability of these larvae to migrate after the fish death. The A. simplex s.s./A. pegreffii larvae proportion found on the anchovy surface is similar to the found in viscera and lower than in muscle, suggesting that most of the larvae migrating to the surface must have come from the visceral package. This confirms the importance of removing fish viscera immediately after capture, for those fish species where this is possible. As both species cause anisakiasis/anisakidosis, these data show a real risk to human health, especially in dishes highly prized in Mediterranean countries prepared with raw or semi-raw anchovies., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: To investigate the prevalence of Anisakis infection in marine fishes in Eastern Fujian Fishing Ground of Fujian Province, so as to provide insights into the development of the anisakiasis control strategy., Methods: Marine fish samples were randomly collected from Jiaocheng District, Fuding City and Xiapu County around Eastern Fujian Fishing Ground in Fujian Province from September to December, 2021. All fishes were dissected, and the abdominal contents were collected. Parasites were sampled under a stereomicroscope and the Anisakis species were identified through morphology. The prevalence and intensity of Anisakis infections were calculated., Results: A total of 177 marine fishes belonging to 24 species were dissected, and Anisakis was detected in 73 marine fishes (41.2%) belonging to 16 species (66.7%), with a mean infection intensity of 14.3 parasites per fish. High prevalence of Anisakis infection was found in Ilisha elongata (5/5), Miichthys miiuy (3/3), Plectorhynchus cinctus (2/2), Scomberomorus niphonius (12/13), Trichiurus lepturus (19/23), Pennahia argentata (6/11) and Ditrema temmincki (14/27), with mean infection intensities of 9.2, 2.7, 4.5, 10.9, 39.2, 4.5 parasites per fish and 2.1 parasites per fish. The Anisakis larvae were characterized as Anisakis and Hysterothylacium., Conclusions: High prevalence of Anisakis infection is detected in marine fishes in Eastern Fujian Fishing Ground of Fujian Province. The health education pertaining to food health is required to be reinforced to prevent the development of human anisakiasis.

Background: Anisakiasis is a foodborne disease caused by the third-stage larvae (L3) of two species belonging to the genus Anisakis: Anisakis pegreffii and Anisakis simplex sensu stricto. Both species have been the subject of different -omics studies undertaken in the past decade, but a reliable in vitro culture protocol that would enable a more versatile approach to functional studies has never been devised. In nature, A. pegreffii shows a polyxenous life-cycle. It reproduces in toothed whales (final host) and disseminates embryonated eggs via cetacean faeces in the water column. In the environment, a first- (L1) and second-stage larva (L2) develops inside the egg, and subsequently hatched L2 is ingested by a planktonic crustacean or small fish (intermediate host). In the crustacean pseudocoelom, the larva moults to the third stage (L3) and grows until the host is eaten by a fish or cephalopod (paratenic host). Infective L3 migrates into the visceral cavity of its paratenic host and remains in the state of paratenesis until a final host preys on the former. Once in the final host's gastric chambers, L3 attaches to mucosa, moults in the fourth stage (L4) and closes its life-cycle by becoming reproductively mature., Methods: Testing two commercially available media (RPMI 1640, Schneider's Drosophila) in combination with each of the six different heat-inactivated sera, namely foetal bovine, rabbit, chicken, donkey, porcine and human serum, we have obtained the first reliable, fast and simple in vitro cultivation protocol for A. pegreffii., Results: Schneider's Drosophila insect media supplemented with 10% chicken serum allowed high reproducibility and survival of adult A. pegreffii. The maturity was reached already at the beginning of the third week in culture. From collected eggs, hatched L2 were maintained in culture for 2 weeks. The protocol also enabled the description of undocumented morphological and ultrastructural features of the parasite developmental stages., Conclusions: Closing of the A. pegreffii life-cycle from L3 to reproducing adults is an important step from many research perspectives (e.g., vaccine and drug-target research, transgenesis, pathogenesis), but further effort is necessary to optimise the efficient moulting of L2 to infective L3., (© 2023. The Author(s).)

Effects of temperature on development of eggs, recently hatched larvae and L3 larvae of the marine parasitic nematodes Anisakis simplex sensu stricto (s.s.) and A. pegreffii were examined in vitro. The eggs of A. simplex s.s. hatched at 3-25 °C and those of A. pegreffii hatched at 3-27 °C. Days before hatching varied between 2 days at 25 °C and 35-36 days at 3 °C in A. simplex s.s. and between 2 and 3 days at 27 °C and 65 days at 3 °C in A. pegreffii. Hatching rates of A. simplex s.s. were maintained high at temperatures between 3 and 25 °C but decreased to 0% at 27 °C. In contrast, those of A. pegreffii were lowest particularly at 3 °C, but also at 27 °C. The mean 50% survivals of hatched larvae ranged from 5.3 days at 25 °C to 82.3 days at 9 °C in A. simplex s.s., while in A. pegreffii it ranged from 1.2 days at 27 °C to 77.2 days at 9 °C. L3 larvae of A. pegreffii exhibited higher survival rates and activity than those of A. simplex s.s., particularly at 20 and 25 °C. These results suggest that the early stages of A. simplex s.s. are more adapted to lower temperatures whereas those of A. pegreffii are more tolerant to warm environments, which may correspond to their distribution patterns in Japan and Europe., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This study determines the occurrence and molecular identification of nematodes from blue mackerel Scomber australasicus Cuvier (Perciformes: Scombridae), an edible fish from Australian waters. A total of 150 fish were sourced from the New South Wales and Victorian coasts. Nematodes were initially classified morphologically as 10 morphotypes belonging to the families Anisakidae (Anisakis morphotypes I and II, Contracaecum morphotype II, Terranova morphotypes I and II), Raphidascarididae (Hysterothylacium morphotypes IV, VI, VIII, and XIV), and Capillariidae (Capillaria sp.), followed by molecular identification through sequencing of their internal transcribed spacer (ITS-1, 5.8S, ITS-2) region. Anisakis morphotype I was confirmed as A. pegreffii Campana-Rouget & Biocca, 1955 and A. berlandi Mattiucci, Cipriani, Webb, Paoletti, Marcer, Bellisario, Gibson & Nascetti, 2014. Anisakis morphotype II and Contracaecum morphotype II were confirmed as A. physeteris Baylis, 1923 and C. ogmorhini Johnston & Mawson, 1941, respectively. Terranova morphotypes I and II were identified as Pulchrascaris australis Shamsi, Barton & Zhu, 2020 and Euterranova pectinolabiata n. comb. (Shamsi, Barton & Zhu, 2019) Moravec & Justine, 2020, respectively. The specific identification of Hysterothylacium morphotypes IV, VI, and VIII was not possible as no comparable adult Hysterothylacium species sequences were available in GenBank, with the exception of morphotype XIV which was confirmed as H. persicum Shamsi, Ghadam, Suthar, Mousavi, Soltani & Mirzargar, 2016. Seven nematode morphotypes were identified for the first time in Australian blue mackerel. The outcomes of the study provide a basis for future research into the community structure, life cycles, and distribution of nematode species in Australian mackerel and to analyse and clarify their importance for public health., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Reactivity to an anisakis allergen component was examined in three patients with a history of an anisakiasis anaphylaxis. Case 1, a 38-year-old man, allergic symptoms appeared 0.5 hours after ingestion, and the component Ani s 1 and 3 were positive. Case 2, a 44-year-old woman, allergic symptoms appeared 4 hours after ingestion, and components Ani s 3 and 12 were positive. Case 3, a 36-year-old woman, developed allergic symptoms 7 hours after ingestion of fish and shellfish, and tested positive for Ani s 1, 4, and 12. Case 3 reacted strongly to both heated and unheated Anisakis extract, while cases 1 and 2 reacted weakly to heated Anisakis extract. The most common allergen was Ani s 12, followed by Ani s 1, when analyzed in conjunction with existing reports on 10 cases. Anisakis IgE was class 3 or higher in all cases. Analysis of 13 cases showed 2 cases sensitized to Ani s 4 and moderate or higher anaphylaxis, while Ani s 4-sensitized patients were reported to be more likely to develop severe disease. It is possible that the patients sensitized to Ani s 4 need to be careful about the severity of their allergic symptoms.

Anisakids are nematodes responsible for different clinical patterns in humans. The well-known human-infecting Anisakis species include members of the Anisakis simplex (AS) complex. Humans usually contract anisakiasis through ingestion of raw or undercooked seafood containing Anisakis larvae. Once Anisakis has been ingested, patients may develop disease driven directly by Anisakis larvae and/or by allergic reaction due to this nematode. The capability of inducing allergic reactions depends on the expression of specific antigens by nematodes and host factors. This study aims to resume actual knowledge about AS and Anisakiasis with regard to epidemiology, pathophysiology, clinical presentation, diagnosis, and treatment. Particular attention is paid to Anisakis allergens and their cross-reactivity on available diagnostic methods, and defining a diagnostic pathway for Anisakis allergy. Because only a few data are available in the literature about pediatric population, we focus on this group of patients specifically., Competing Interests: The authors have no conflict of interest relevant to this article to disclose.

Infective third-stage larvae (L3) of the marine nematode Anisakis pegreffii cause inflammation and clinical symptoms in humans, their accidental host, that subside and self-resolve in a couple of weeks after L3 die. To characterise the differences in an early immune response of a marine vs. terrestrial host, we stimulated peripheral blood leukocytes (PBLs) of fish (paratenic host) and rat (accidental, human-model host) with A. pegreffii crude extract and analysed PBL transcriptomes 1 and 12 h post-stimulation. Fish and rat PBLs differentially expressed 712 and 493 transcripts, respectively, between 1 and 12 h post-stimulation (false discovery rate, FDR <0.001, logFC >2). While there was a difference in the highest upregulated transcripts between two time-points, the same Gene Ontologies, biological processes (intracellular signal transduction, DNA-dependent transcription, and DNA-regulated regulation of transcription), and molecular functions (ATP and metal ion binding) were enriched in the two hosts, showing an incrementing dynamic between 1 and 12 h. This suggests that the two distinct hosts employ qualitatively different transcript cascades only to achieve the same effect, at least during an early innate immunity response. Activation of later immunity elements and/or a combination of other host's intrinsic conditions may contribute to the death of L3 in the terrestrial host., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hrabar, Petrić, Cavallero, Salvemini, D’Amelio and Mladineo.)

In recent years, a drop in the condition of the European sardine has been observed. Although several causes have been attributed to this issue, as overfishing and climate change, little is known about the link between ascaridoid nematode parasitisation and fish status. In this study, sardines were obtained from four fishing grounds along the Mediterranean (Alboran, Northern Spain, Northern Adriatic, and Aegean), and one location in the Atlantic Ocean (Southern Portugal). After analysing individual fish body condition (by direct tissue fat content measurements and condition indices), and reproductive status (by a detailed gonadal examination) throughout the entire annual cycle, ascaridoids were recognised by combining naked eye and UV-press method along flesh, viscera, and gonads. Afterwards, sequence analysis of the rDNA internal transcribed spacers region (ITS) and the mtDNA cox2 gene were used to identify and characterise the different species of ascaridoids from the fish host in the localities throughout the seasons. The main species found along different areas was Hysterothylacium aduncum , present in the Northern Adriatic (prevalence of 7.6%, mean intensity 1.700), the Atlantic (7.5%, 3.889), and the Northern Spain (3.9%, 1.600). Moreover, few individuals of Anisakis simplex (s.s.) and A. pegreffii were observed in the Atlantic (1.7% and 0.8%, respectively), and the latter species was also found in the Adriatic stock (0.8%). All ascaridoid specimens were found in viscera. Obtained results seem to indicate that in stocks with medium sizes, small variations in length are related to parasite intensity. This study highlights the importance of seasonal parasitological analyses at stock level and, especially, in capital breeders, as relationships between condition and reproduction parameters and parasitism are conditioned by seasonality., Competing Interests: The authors declare no conflict of interest/competing interest., (© 2022 The Authors.)

Northeast Arctic cod, saithe and haddock are among the most important fisheries resources in Europe, largely shipped to various continental markets. The present study aimed to map the presence and distribution of larvae of parasitic nematodes in the Anisakidae family which are of socioeconomic and public health concern. Fishes were sourced from commercial catches during winter or spring in the southern Barents Sea. Samples of fish were inspected for nematodes using the UV-press method while anisakid species identification relied on sequencing of the mtDNA cox2 gene. Anisakis simplex (s.s.) was the most prevalent and abundant anisakid recorded, occurring at high infection levels in the viscera and flesh of cod and saithe, while being less abundant in haddock. Contracaecum osculatum (s.l.) larvae, not found in the fish flesh, showed moderate-to-high prevalence in saithe, haddock and cod, respectively. Most Pseudoterranova spp. larvae occurred at low-to-moderate prevalence, and low abundance, in the viscera ( Pseudoterranova bulbosa ) and flesh ( Pseudoterranova decipiens (s.s.) and Pseudoterranova krabbei ) of cod, only 2 P. decipiens (s.s.) appeared in the flesh of saithe. Body length was the single most important host-related factor to predict overall abundance of anisakid larvae in the fish species. The spatial distribution of Anisakis larvae in the fish flesh showed much higher abundances in the belly flaps than in the dorsal fillet parts. Trimming of the flesh by removing the belly flaps would reduce larval presence in the fillets of these gadid fish species by 86–91%.

Introduction: Anisakiasis is a human parasitic disease caused by the consumption of raw fish or shellfish containing larvae of the Anisakidae family. It is currently considered an emerging disease of public health interest., Objective: To identify the presence of larvae of the Anisakidae family in samples of frozen raw fish fillets intended for human consumption in markets in Medellín and its metropolitan area in Antioquia, Colombia., Materials and Methods: A cross-sectional study was carried out, in which larvae of the Anisakidae family were detected and identified in frozen raw fish fillets from three representative markets in Medellín and its metropolitan area. A total of 384 ready for consumption fillets were analyzed (197 sawfish, 137 salmon, 37 tuna, and 13 hake), using the pressing and ultraviolet light method. Taxonomic keys were used to identify the collected parasites and to establish its genus. Conventional PCR and Sanger sequencing was performed to determine the species., Results: Four larvae were found in 4 of the 384 (1.04%) fillets (CI95% 1.04 ± 1.01%). The species of fish in which the larvae were found was sawfish (Scomberomorus spp.) and the genus and species of the larvae was established as Anisakis pegreffii., Conclusions: According to the study, the presence of Anisakis parasites in frozen raw fish fillets in the influence area is evident.

Spotted flounder ( Citharus linguatula L.) caught in the Gulf of Cadiz (area FAO 27 ICES IXa) were examined for Anisakis larvae and to assess the possible risk of anisakiasis in humans through consumption of this fish. Larvae of the genera Anisakis and Hysterothylacium were identified in the analysis of 128 purchased fish specimens. All Anisakis larvae corresponded to type I. Molecular analysis showed the presence of A. pegreffii , A. simplex s.s., and recombinant genotype between the two. The prevalence of Anisakis was 9.4% with a mean intensity of 1.42, while for Hysterothylacium the values were 12.5% and 1.06. The length and weight of the fish, but not Fulton's condition factor, varied significantly between infected and uninfected fish. The prevalence of Anisakis increased with fish length, with no fish parasitized with Anisakis measuring less than 15.5 cm (2-2.5 years old), which is probably related to the reported dietary change of these fish at around 2 years of age. Fish not parasitized with any of these nematodes showed positive allometric growth, while those parasitized only with Anisakis showed negative allometric growth. When comparing both groups including only fish ≥ 15.5 cm (the smallest size of Anisakis -infected fish), the difference is shown to be statistically significant ( p = 0.01), suggesting that Anisakis infection of spotted flounder negatively affects fish growth even when parasite intensity is low, which may have important economic repercussions. Finally, the low prevalence and, above all, intensity of Anisakis in these fish, as well as the habit of consuming this fish fried in oil in our geographical area, means that the risk of acquiring anisakiasis through consumption of this fish is low.

Anisakis spp. (Nematoda, Anisakidae) are parasites known by their economic and health impacts, as their L3 larval stages infect a variety of fish species, many of them commercial species, sometimes causing zoonotic episodes due to consumption of raw or undercooked fish. The aim of this study is to determine the infection process and the potential impact of A. simplex s.l. L3 on gilthead seabream (Sparus aurata L.), one of the most important fish species in Mediterranean aquaculture, by periodic histological monitoring of the infection process. For this, fish were experimentally infected with A. simplex s.l. L3 and periodically analysed for L3 larvae, collecting samples at different time points (hours post ingestion, hpi): 3, 6, 12, 18, 24, 36, 48, 72, 96, 120, 144, 168 and 192, up to 6 months post infection (mpi). All samples were observed under a stereomicroscope and later fixed for histological examination. A. simplex s.l. L3 were only found on the visceral surface and mesenteric tissue, but never free or encapsulated in muscle. Chronological events were found to occur faster than those reported in previous studies. They were first observed 6 hpi in the coelomic cavity, being present up to 48 hpi. While the earliest evidence of fibrocytes surrounding A. simplex s.l. L3 larvae were observed at 18 hpi, complete spiral encapsulation occurred by 72 hpi. Alive parasites were observed up to 6 mpi. Although the infection of gilthead seabream by Anisakis spp. larvae is feasible, it seems unlikely, especially in aquaculture given the hygienically controlled feeding systems. In the event of infection, the transmission would be unlikely due to the poor condition in which specimens of Anisakis spp. are found. Furthermore, since no larvae were detected in the fish's muscle, human infection seems improbable., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Reconstruction of eyelid margin defects following resection of upper-eyelid skin malignancies is typically performed using a lower eyelid switch flap, including eyelash reconstruction. However, a subsequent procedure for flap separation, and prolonged swelling of the flap may occur as a complication., Objective: The authors performed anterior lamellar reconstruction using a sliding flap with excess upper eyelid skin, a procedure that is a less invasive and simpler., Materials and Methods: The authors performed anterior lamellar reconstruction using a sliding flap in 7 patients with full-thickness upper eyelid margin defect after skin cancer resection. The mean age of the patients was 76.0 years. The horizontal width of the defect in our cohort ranged from 11 to 25 mm and the vertical width ranged from 5 to 10 mm., Results: All the flaps and mucosal grafts were well taken, and none of the patients complained of lack of eyelash reconstruction. However, eyelid margin irregularity, possibly due to flap or mucosal graft contraction, was observed in 2 patients, and 1 patient developed keratoconjunctivitis., Conclusions: The sliding flap technique is a minimally invasive and simple procedure for wide eyelid margin reconstruction. However, 2 patients developed eyelid margin irregularities and 1 patient developed keratoconjunctivitis. Those complications might have occurred owing to the condition of posterior lamellar reconstruction. Therefore, for the successful use of a sliding flap for anterior lamellar reconstruction, the form of the eyelid edge and the choice of posterior lamellar reconstruction are key considerations. In future, we plan to establish a better reconstructive technique by accumulating more evidence., Competing Interests: The authors report no conflicts of interest., (Copyright © 2022 by Mutaz B. Habal, MD.)

Background : Anisakis spp. third-stage larvae (L3) are the causative agents of human zoonosis called anisakiasis. The accidental ingestion of L3 can cause acute and chronic inflammation at the gastric, intestinal, or ectopic levels. Despite its relevance in public health, studies on pathogenetic mechanisms and parasite-human interplay are scarce. The aim of this study was to investigate the human inflammatory response to different Anisakis vehicles of pathogenicity. Methods : Human colorectal adenocarcinoma (Caco-2) cells were exposed to Anisakis L3 (the initial contact with the host), extracellular vesicles (EVs, Anisakis -host communication), and crude extract (CE, the larval dying). The protein quantity and gene expression of two pro-inflammatory cytokines (IL-6 and IL-8) were investigated using an ELISA test (6 h and 24 h) and a qReal-Time PCR (1 h, 6 h, and 24 h), respectively. Results : The L3 and EVs induced a downregulation in both the Il-6 and Il-8 gene expression and protein quantity. On the contrary, the CE stimulated IL-6 gene expression and its protein release, not affecting IL-8. Conclusions : The Caco-2 cells seemed to not react to the exposure to the L3 and EVs, suggesting a parasite's immunomodulating action to remain alive in an inhospitable niche. Conversely, the dying larva (CE) could induce strong activation of the immune strategy of the host that, in vivo, would lead to parasite expulsion, eosinophilia, and/or granuloma formation.

IL-7 is a crucial factor for the development of lymphocytes, and it is absolutely necessary for γδ T cells. Mice deficient in L-7 have a deficit of B and αβ T lymphocytes, and an absence of mature γδ TCR cells. IL-7 is essential for the survival, development and maturation of Schistosoma sp., although its production is associated with protection against intestinal helminths. The presence of anti-Anisakis simplex antibodies, especially IgA, is related to a lower frequency in CD3 + CD56 + αβ + lymphocytes and all subpopulations of γδ T cells. In this work, the relationship of IL-7 with humoral and cellular responses against A. simplex in 100 healthy subjects was studied. We have found significantly higher IL-7 levels in anti-A. simplex IgA-positive subjects (p < 0.001). The positivity of anti-A. simplex IgA was associated with a significant reduction in the frequency of CD3 + αβ+ (p < 0.01), CD3 + CD4 + αβ+, CD3 + CD8 + αβ+, CD3 + CD56 + αβ+, CD3 + γδ+, CD3 + CD4-CD8-γδ+ and CD3 + CD56 + γδ+ (p < 0.05) cells. In the case of NKT cells, this same phenomenon was also associated with IgE positivity. There was a weak inverse correlation (Spearman) of IL-7 levels with the frequencies of CD3 + CD4 + αβ+ (-0.125, p = 0.047), CD3 + CD8 + αβ+ (-0.204, p = 0.032), CD3 + CD56 + αβ+ (-0.247, p = 0.007), CD3 + γδ+ (-0.267, p = 0.007), CD3 + CD4-CD8-γδ+ (-0.266, p = 0.003), and CD3 + CD8 + γδ + (-0.302, p = 0.002) cells. The role of NKT cells in the anti-A. simplex response was confirmed and an association between IL and 7 levels and specific antibodies, especially IgA, was demonstrated. The higher production of IL-7 would represent a compensatory mechanism in response to the reduction in lymphocyte populations associated with the response against this parasite., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)