Human Anisakiasis Research Articles

Prevalence of Anisakis Larvae in Different Fish Species in Southern Albania: Five-Year Monitoring (2016–2020)

Anisakidae are nematodes that commonly parasitize in the coelomic cavity and viscera of several fish species. They can be found in flesh, which is why they have an important economic and public health impact. The aim of the current work was to assess the presence and prevalence of Anisakis larvae in fish species caught in the coastal area of the Karaburun Peninsula in Vlora Bay (Albania). A total of 856 of wild teleosts and 219 specimens of farmed fish were collected over a 5-year period (from 2016 to 2020). The results showed that out of a total of 1075 analyzed samples, 361 (33.58%) were parasitized with L3 larvae. In particular, only Solea vulgaris returned negative results, while Sparus aurata, Dicentrarchus labrax, and Sardinella aurita showed the lowest prevalence (4.55%, 9.17%, and 10.53%, respectively) and mean abundance (0.84, 1.19, and 0.92, respectively). Conversely, Scomber japonicus and Scomber scombrus showed the highest prevalence (74.07% and 68.00%, respectively) and mean abundance (188.24 and 249.82, respectively). The data suggest that the coastal area of the Karaburun Peninsula (southern Albania) may be a high-risk area for zoonotic diseases, and the consumption of raw or undercooked fish caught in the Vlora district could result in the acquisition of human anisakiasis. For these reasons, it is necessary to improve the surveillance plan.

Molecular identification and epidemiological data of Anisakis spp. (Nematoda: Anisakidae) larvae from Southeastern Pacific Ocean off Peru.

The objective of this study is to determine the infection status of nematode larvae and record epidemiological molecular data in commercial fish from the southeast Pacific off the central coast of Peru. Anisakiasis is a fish-borne zoonosis caused by Anisakis larvae, parasites of relevance in the fishery resources that have negative impact on public health. Between January 2012 to December 2014, 345 specimens of four fish species (Trachurus symmetricus murphyi, Scomber japonicus peruanus, Merluccius gayi peruanus and Seriolella violacea) were examined for Anisakis sp. larvae. A total of 997 Anisakis sp. larvae were found in the body cavity of 196 fish (total prevalence 53.7%, total mean intensity 5.08). After morphological analysis, 958 (96.08%) larvae were identified as Type I and 39 (3.92%) as Type II. Specimens were identified by molecular analysis of the mitochondrial cytochrome c oxidase subunit II (cox2) gene, confirming that A. pegreffii is the predominant species and the most important agent of human anisakiasis off the Peru Central Coast. In addition, we revealed the occurrence of A. physeteris (s.l.) in S. japonicus peruanus (P = 18.0%; MI = 2.17).Therefore, the results obtained in the present study improve the knowledge of the occurrence of Anisakis species in the commercial fish from the Southeastern Pacific Ocean, highlighting the importance of considering a potential hazard for humans and the necessity of further research in other fishes of greater preference by the Peruvian population.

Hybrid Genotype of Anisakis simplex (s.s.) and A. pegreffii Identified in Third- and Fourth-Stage Larvae from Sympatric and Allopatric Spanish Marine Waters.

Simple SummaryThe nematode species Anisakis simplex sensu stricto (s.s.) and Anisakis pegreffii are wormlike parasites found in commonly consumed fish and are the main cause of human anisakiasis. Outwardly, the two nematodes are extremely similar and difficult to distinguish, especially in their larval forms. Genetic analysis has discovered the existence of a hybrid between these two “sibling species”, but its identification is a controversial matter, as results differ according to the specific region of the DNA analysed. The aim of our work was to confirm the presence of this hybrid genotype in fish off the Spanish coast and to obtain fourth-stage larvae in the laboratory to study if different genotypes are associated with different growth behaviour. Our results confirm that hybrid genotypes can be overestimated if identification is based on a particular molecular marker. We also obtained fourth-stage larvae with a hybrid genotype, which has not been reported previously. These findings are valuable for the taxonomic classification of Anisakis species, and for further epidemiological and biomedical research.The sibling species Anisakis simplex (s.s.) and Anisakis pegreffii are parasites of marine mammals and fish worldwide and the main causative agents of human anisakiasis. In sympatric areas, a hybrid genotype between the two species has been identified, mainly in third-stage larvae, but rarely in fourth-stage and adult forms. The aim of this study was to confirm the presence of hybrid genotypes in larvae parasitizing fish caught in sympatric and allopatric Spanish marine waters, the North-East Atlantic and West Mediterranean, respectively, and to study possible differences in the growth behaviour between genotypes. Of the 254 molecularly analysed larvae, 18 were identified as hybrids by PCR-RFLP analysis of the rDNA ITS region, 11 of which were subsequently confirmed by EF1 α-1 nDNA gene sequencing. These results therefore indicate an overestimation of hybrid genotypes when identification is based only on the ITS region. We also report the detection of a hybrid specimen in a host from the West Mediterranean, considered an allopatric zone. Additionally, fourth-stage larvae with a hybrid genotype were obtained in vitro for the first time, and no differences were observed in their growth behaviour compared to larvae with A. simplex (s.s.) and A. pegreffii genotypes.

Helminth Parasites of Caranx hippos (Crevalle Jack) and Sardinella maderensis (Madeiran Sardinella) from Two Fishing Settlements In Okrika, Rivers State, Nigeria

Fish is an important dietary component as a source of high quality protein, and parasitic infections can result in production and economic losses. Therefore, this study was aimed at investigating the helminth parasites of Caranx hippos and Sardinella maderensis of Okrika. Fish samples were purchased from fishers between May and June, 2019 from two locations: Angalabia and Sara landing ports, Okrika. Fifty specimens of each species were dissected using standard methods, and parasites were identified using appropriate keys. Parasite ecological parameters were computed. Two parasite species were identified, Lethacotyle sp. (monogenea) and Anisakid larvae (nematode). Twenty-six specimens of Caranx hippos were infected with Lethacotyle sp., and eight with Anisakid larvae. Only one specimen of S. maderensis was infected with Anisakid larvae; others were un-infected. Examination showed a prevalence of 52.0% and 16.0% infection of Lethacotyle sp. and Anisakid larvae, respectively, in Caranx hippos, and 2.0% prevalence of Anisakid larvae in Sardinella maderensis. The preferred organs for parasite infection were the gills (Lethacotyle sp.) and intestines (Anisakid larva) of infected fish. The recovery of Anisakid larvae at a prevalence of 16% indicates that anisakiasis could be an emerging disease in Okrika. It is recommended that further research be conducted into the molecular characterization of Lethacotyle sp. of C. hippos of Okrika. It is also important to properly identify and characterize the Anisakid species, and investigate the incidence of human anisakiasis inOkrika and other coastal cities of Rivers State, Nigeria.
 Keywords: Lethacotyle, Anisakid larva, anisakiasis, Caranx, Sardinella, Parasites, Marine, Okrika, Nigeria

Shotgun Proteomics for L3 and L4 Anisakis simplex Development Stages.

Anisakis simplex s.s. is a parasitic nematode that causes anisakiasis in humans. L3 stage larvae, which are present in many fish species and cephalopods all over the globe, might be consumed and develop occasionally into the L4 stage but cannot reproduce. Anisakiasis is an emerging health problem and economic concern. In recent years, proteomic methods have gained greater acceptance among scientists involved in parasitology and food science. According to that, here, we present tandem mass tag (TMT)-based shotgun proteomics to define differences in proteomic composition between L3 and L4 development stages of A. simplex s.s.

First morphological and molecular identification of third-stage larvae of Anisakis typica (Nematoda: Anisakidae) from marine fishes in Vietnamese water.

Anisakid nematodes are parasites of cetaceans, their larval stages live in marine fishes. The third-stage larvae of some Anisakis species are also the etiological agents of human anisakiasis caused by consumption of raw or undercooked infected fish. Thus, identification of Anisakis larvae at the species level is crucial for their ecology and epidemiology. In Vietnam, although Anisakis larvae have been reported, they have not been identified to the species level. The aim of this study was, therefore, to identify third-stage larvae of Anisakis collected from marine fishes in Vietnamese water, based on morphological characteristics and molecular analysis. All Anisakis larvae found in this study were morphologically similar to each other and identical to Anisakis typica. In addition, molecular analysis based on ITS1-5.8S-ITS2 sequences confirmed them as A. typica. Vietnamese A. typica population was genetically close to those from Asian countries and Australia. The third-stage larvae of A. typica were collected from eight fish species from three localities in the South of Vietnam. Among them, seven were recorded as new intermediate hosts of A. typica. This is the first identification of A. typica larvae in Vietnamese water with records of new fish hosts.

Intra-vitamlarval migration of Anisakis pegreffii in live spotty-bellied greenling, Hexagrammos otakii, at different water temperatures

The present study investigated the intra-vitam larval migration of Anisakis pegreffii, which is naturally infected, from the intestine to the viscera and the musculature of live spotty-bellied greenling, Hexagrammos otakii, under the different seawater temperatures. The results showed that the increase in the ratio of larval migration from the intestine to the viscera correlated with the increase in the water temperature in the aquarium. The intra-vitam larval migration ratios from the intestine to the viscera were 60% (63/105) at 15°C, 1.64% (3/183) at 12°C, 1.16% (2/172) at 9°C and 1.42% (2/141) in the control group (7.5°C). The free larvae showed no, slight and strong movement at 9°C, 12°C and 15°C, respectively, in saline water, and the larvae were present in a coiled state at 9°C and 12°C. The results suggest that temperature plays an important role in the intra-vitam larval migration of A. pegreffii. In addition, the free larval behavior of A. pegreffii in vitro increased with increase in temperature. Therefore, to prevent human anisakiasis caused by the consumption of raw fish, it is important that the temperature of the aquarium must be maintained below 12°C, which is the critical temperature zone.

Parasitic Anisakid Nematode Isolated from Stranded Fraser’s Dolphin (Lagenodelphis hosei Fraser, 1956) from Central Philippine Waters

Cetaceans, including dolphins, serve as definitive hosts of zoonotic anisakid nematodes, which are important etiological agents for human anisakiasis and allergy-associated health risks. With limited knowledge of these zoonotic parasites from the marine environment in the Philippine waters, the stranding of a Fraser’s dolphin (Lagenodelphis hosei Fraser, 1956) off the central Philippines made it possible to identify the worm species isolated from its gut. Parasitological examinations were carried out using morphological and molecular tools. Morphologically, the SEM and LM data revealed that the specimens belong to the genus Anisakis of the Type 1 group. Molecularly, PCR-RFLP results of the ITS region generated only a single fragment pattern on all worm samples corresponding to the reported molecular keys for A. typica. Further sequence and phylogenetic analyses of both ITS rDNA and mtDNA COX2 genes confirmed the anisakid nematodes’ identity as A. typica. The molecular data obtained in this study support previous findings on the possible existence of local variants of A. typica in this region.

Probable association between Anisakis infection in the muscle of skipjack tuna (Katsuwonus pelamis) and human anisakiasis in Tokyo, Japan

Anisakiasis is a gastrointestinal disease caused by parasitic anisakid nematodes, mainly Anisakis simplex sensu stricto (A. simplex). Anisakiasis is prevalent in Japan and approximately 40% of anisakiasis cases in Tokyo occur through the consumption of raw or marinated mackerel. However, in 2018, there was a sudden increase in the number of the food poisoning cases in Tokyo caused by consumption of skipjack tuna (Katsuwonus pelamis). Therefore, we investigated anisakiasis cases resulting from ingestion of skipjack tuna in Tokyo, and surveyed the presence of Anisakis larvae in skipjack tuna in 2018 and 2019. Nineteen samples from 15 patients (13 in 2018 and 2 in 2019) with anisakiasis surely caused by ingestion of skipjack tuna were all identified as A. simplex. The higher mean abundance of Anisakis simplex larvae in skipjack tuna muscle in May 2018 (1.30; 13 larvae/10 fishes) compared to that in the other periods was regarded as a contributing factor in the increase in anisakiasis cases by ingesting skipjack tuna in 2018. To verify whether Anisakis larvae migrate from the visceral organs to the muscle during the period from fishing on the boat until processing for sale, the number of Anisakis larvae in skipjack tuna caught from August to November 2018 was investigated by removing the visceral organs at three different timings, i.e., immediately after catching, after landing, and after transport to the laboratory. Anisakis larvae were detected in the muscle irrespective of the timings at which visceral organs were removed. All larvae from the muscle were detected only from the ventral part and were identified as A. simplex. We thus consider that avoiding raw consumption of the ventral muscle should be an effective measure to prevent anisakiasis.

World-wide prevalence of Anisakis larvae in fish and its relationship to human allergic anisakiasis: a systematic review.

The infective stage of Anisakidae nematodes responsible for allergic reactions in humans is found in a variety of edible fish and cephalopods. The identification of geographical regions that are high risk for infected seafood may help prevent allergic reactions in humans. Despite an abundance of published literature which has identified anisakid larvae in an array of edible seafood as well as scattered reports of human allergic anisakiasis, the relationship between the two has not been fully explored. Therefore, a systematic spatio-temporal study was conducted to determine the prevalence of Anisakis spp. in fish from January 2000 to August 2020 firstly to explore the relationship between fish infection and cases of allergic anisakiasis and secondly to use fish infection data to map potential allergic anisakiasis 'hot spots'. A systematic literature search for original English text articles was conducted through search engines, Web of Science, Scopus, PubMed, Science Direct and Google Scholar. Out of 3228 articles which describe anisakid infection in fish, 264 were used for data extraction. Of 904 articles describing allergic anisakiasis, 37 were used for data extraction. A qualitative summary of the extracted data was performed using equal interval method (ArcMap software) in order to compare the global distribution of Anisakis-infected fish. Of the 152-identified fish hosts, five families were most commonly infected with Anisakis spp. These included Lophiidae (86.9%), Trichiuridae (77.05%), Zeidae (70.9%), Merlucciidae (67.8%) and Gadidae (56.8%). The hot spot areas for allergic anisakiasis were North and northeast of Atlantic Ocean, southwest of USA, west of Mexico, south of Chile, east of Argentina, Norway, UK and west of Iceland (confidence 99%). The highest rate of allergic anisakiasis was in Portugal and Norway with the prevalence rate of 18.45-22.50%. Allergologists should consider allergic anisakiasis as a public health issue particularly in high-risk countries where high prevalences in fish have been demonstrated.

Investigation of Anisakis larvae in different products of ready-to-eat fish meat and imported frozen fish in Turkey

Globalization opens new market areas and affects food consumption habits, resulting in rapid and remarkable cultural change. Food habits such as consumption of raw fish meat have become popular, resulting in increased risk of emerging infectious diseases. Anisakis simplex sensu stricto (s.s) and A. pegreffii are the most common and important fish-borne zoonotic nematodes responsible for human anisakiasis, which occurs through the consumption of raw or undercooked fish as well as cooked fish due to their heat-stable allergens. Here, we investigated the prevalence, intensity, and abundance of Anisakis larvae in imported fish and ready-to-eat local fish products in Turkey. A total of 205 ready-to-eat fish products, 100 imported frozen Atlantic salmon (Salmo salar) fillets, and 100 imported frozen whole Atlantic mackerel (Scomber scombrus) were sampled from supermarkets, sushi restaurants, and fish markets. All samples were individually examined using a pepsin digestion technique. In total, 602 Anisakis type I larvae were recovered from 98/100 mackerel. No larvae were found in ready-to-eat products or frozen Atlantic salmon fillets. Overall, 8.8% of the larvae were found in the muscle tissue. The overall mean intensity and abundance of infection in mackerel were 6.14 and 6.02, respectively. The larvae were molecularly identified and their phylogenetic relationships with the relevant Anisakis sequences in GenBank were investigated. For this purpose, a subsample of randomly selected 100 Anisakis larvae were analyzed with PCR-RFLP of the ITS region. The larvae were identified as A. simplex (s.s.) (n = 87) and hybrids (n = 13). ITS and cox2 gene regions of all hybrids and randomly selected 50 A. simplex (s.s.) larvae were sequenced for species confirmation and phylogenetic analyses. No intraspecific nucleotide variation was found among the ITS sequences of either species. Seven and three haplotypes, respectively, were identified for A. simplex (s.s.) and hybrid species according to DNA polymorphism of the cox2 gene. Hybrids in our study clustered within the common A. simplex (s.s.) clade in the cox2 phylogenetic tree indicating the dominance of A. simplex (s.s) in the catching area of Atlantic mackerel. Consequently, our study indicates high occurrence of A. simplex (s.s.) larvae with an overall 98.0% prevalence in imported Atlantic mackerel, and highlights the importance of these fish as potential reservoirs for human allergic anisakiasis in Turkey and possibly in other countries.

Analysis of Ani s 7 and Ani s 1 allergens as biomarkers of sensitization and allergy severity in human anisakiasis

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.

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

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

ANTIPROLIFERATIVE EFFECT OF ANISAKIS SIMPLEX RUDOLPHI, 1809 (DUJARDIN, 1845) L3 EXTRACT ON VERO LINE CELLS IN VITRO

The effect of a sterile crude extract from the frozen Anisakis simplex Rudolphi, 1809 (Dujardin, 1845) third stage larvae on the transplantable culture of mammalian cells is described. At the first time fibroblasts cells line Vero (from African green monkey, kidney) obtained from the «BioloT», St Petersburg, Russia, on a nutrient media EMEM were used. In the in vitro conditions a significant antiproliferative action of crude somatic extract expressed by complete destruction of the monolayer of fibroblast-like cells after 48 hours of cultivation is proven. Therefore, our results can be regard as a manifestation of a pronounced inhibitory effect of the studied extract on the proliferation of Vero culture, which is of considerable interest in the field of cell technology and Parasitology and may be use as the basis for subsequent experiments to study the pathogenesis and prevention of anisakiasis in humans and animals.

Occurrence and molecular identification of Anisakis larval type 1 (Nematoda: Anisakidae) in marketed fish in Egypt.

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 (mt cox1)gene revealed that all larvae isolated from Atlantic herring and Mediterranean horse mackerel belonged to Anisakis simplex sensu stricto with prevalence of 87.1% and 83.3%, respectively, whereas Atlantic mackerel harbored Anisakis typica 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 possiblyin other coastal countries.

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

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

Anisakid Larvae from Anchovies in the South Coast of Korea.

Anisakiasis (anisakidosis) refers to a foodborne zoonosis caused by ingesting raw or undercooked marine fish or cephalopods infected with anisakid larvae. The present study was performed to investigate the prevalence of anisakid larvae in anchovies (Engraulis japonica) purchased from 2 local markets in Gyeongsangnam-do, the Republic of Korea (=Korea), during 2018–2019. Anchovies were transported to our laboratory and examined by pepsin-HCl artificial digestion technique followed by microscopic observations and molecular analyses. The overall prevalence of anisakid larvae was 19.5% (39/200), from which a total of 51 larvae (av. 1.3 larvae/infected anchovy) were recovered. Sequencing of the larvae targeting the ITS region, including ITS1, 5.8S rRNA, and ITS2 genes confirmed the species of larvae as Anisakis pegreffii (54.9%; 28/51), Hysterothylacium sinense (23.5%; 12/51), and Hysterothylacium aduncum (21.5%; 11/51). The results suggested that anchovies could be a potential source of human anisakiasis in Korea.

Comparison of risk factors between human intestinal and gastric Anisakiasis

BackgroundLittle is known in the difference of host factors between intestinal and gastric anisakiasis. The aim of this study was to investigate the associated factors of intestinal anisakiasis in patient's characteristics and the subsequent variation compared to gastric anisakiasis. MethodsAt St. Luke's International Hospital in Tokyo, Japan, a retrospective cohort study was conducted from April 2004 to June 2017. All adult patients who were clinically diagnosed as anisakiasis based on Computed Tomography (CT) scan or endoscopy were included, and anti-Anisakis antibodies (IgG and IgA) were measured for serological validation of anisakiasis, strengthen the diagnosis. Anisakiasis was categorized as either intestinal or gastric depending on its affected site. We compared patients' demographics, social history, and physical and laboratory findings between those with intestinal and gastric anisakiasis by bivariate analyses, followed by multivariate analyses. ResultsA total of 302 patients were included in this study, where the mean age (SD) was 46.5 (14.4) and 66.6% were male. Ninety-two patients (30.5%) had intestinal anisakiasis. Multivariate regression revealed that patients with intestinal anisakiasis were more 45 years old or older (odds ratio (OR) 3.45, 95% confidence interval (CI): 1.53–7.69), male (OR 2.70, 95% CI: 1.20–6.25) and regular alcohol drinker. In terms of the physical and laboratory findings, patients with intestinal anisakiasis had greater heart rate (OR 2.86, 95% CI: 1.33–6.25), higher total protein (OR 2.86, 95% CI: 1.16–6.67), and higher C-reactive protein (CRP) (OR 11.1, 95% CI: 3.03–33.3). ConclusionsOlder males who were regular alcohol drinkers were associated with intestinal anisakiasis, and often heart rate, total protein, and CRP were elevated compared to those of patients with gastric anisakiasis.

Genome-wide analysis of Anisakis simplex sensu lato: the role of carbohydrate metabolism genes in the parasite’s development

Anisakis simplex sensu lato is a parasitic nematode which can cause gastric symptoms and/or allergic reactions in humans who consume raw and undercooked fish. Anisakiasis poses a growing health problem around the globe because it causes non-specific symptoms and is difficult to diagnose. This genome-wide study was undertaken to expand our knowledge of A. simplex s.l. at the molecular level and provide novel data for biological and biotechnological research into the analyzed species and related nematodes. A draft genome assembly of the L3 stage of A. simplex s.l. was analyzed in detail, and changes in the expression of carbohydrate metabolism genes during the parasite's life cycle were determined. To our knowledge, this is the first genome to be described for a parasitic nematode of the family Anisakidae to date. We identified genes involved in parasite-specific pathways, including carbohydrates metabolism, apoptosis and chemo signaling. A total of 7607 coding genes were predicted. The genome of A. simplex s.l. is highly similar to genomes of other parasitic nematodes. In particular, we described a valuable repository of genes encoding proteins of trehalose and glycogen metabolism, and we developed the most comprehensive data set relating to the conversion of both saccharides which play important roles during the parasite's life cycle in a host environment. We also confirmed that trehalose is synthesized at the expense of glycogen. Trehalose anabolism and glycogen catabolism were the predominant processes in stages L4 and L5, which could confirm our and other authors’ previous reports that trehalose is synthesized at the expense of glycogen. The A. simplex s.l. genome provides essential data for post-genomic research into the biology of gastrointestinal and allergic anisakiasis in humans and the biology of other important parasitic helminths.

Anisakis Nematodes in Fish and Shellfish- from infection to allergies.

Anisakidosis is a zoonotic parasitosis induced by members of the family Anisakidae. The anisakid genera includes Anisakis, Pseudoterranova, Hysterothylacium and Contracaecum. The final definitive hosts of these nematodes are marine mammals with a complex life cycle. These nematode parasites use different crustaceans and fish species as intermediate or paratenic hosts and humans are accidental hosts. Human anisakiasis, the infections caused by members of the genus Anisakis, occurs, when seafoods, particularly fish, contaminated with the infective stage (third stage larvae [L3]) of this parasite, are consumed. Pseudoterranovosis, on the other hand is induced by members of the genus Pseudoterranova. These two genera of anisakids have been implicated in human disease globally. There is a rise in reports of gastro-intestinal infections accompanied by allergic reactions caused by Anisakis simplex and Anisakis pegreffii. This review provides an update on current knowledge on Anisakis as a food-borne parasite with special focus on the increasingly reported diversity of fish and crustacean hosts, allergens and immunological cross-reactivity with invertebrate proteins rendering this parasite a significant public health issue.