Morphological and molecular characterization of Hysterothylacium larval morphotypes (Nematoda: Raphidascarididae) infecting edible marine fish in the Black Sea.

The morphological and molecular identification of Hysterothylacium larval morphotypes in the Black Sea remains unresolved and incomplete. The aim of current study was to provide a detailed morphological identification with rDNA whole ITS (ITS‒1, 5.8S subunit, ITS‒2) and mtDNA cox2 sequences data of Hysterothylacium larval morphotypes infecting four commonly edible marine fish species, including European anchovy, Engraulis encrasicolus (L.), horse mackerel, Trachurus trachurus (L.), whiting, Merlangius merlangus (L.), and red mullet, Mullus barbatus ponticus (E.) in the Black Sea (FAO fishing area 37.4.2). Hysterothylacium larval morphotypes were morphologically classified, followed by whole ITS and cox2 sequencing. Four Hysterothylacium larval morphotypes, III, IV, VIII, and IX, are described based on morphological and molecular data. The present study provides the first study reporting whole ITS and cox2 sequences for Hysterothylacium larval morphotypes III, IV, VIII and III, IV, VIII, IX, respectively, in the Black Sea. Here, we offer a foundation for future research on the distribution, morphologic and molecular identification of Hysterothylacium larval morphotypes infecting edible some marine fish in the Black Sea.

In situ evidence of the role of Crangon crangon in infection of cod Gadus morhua with nematode parasite Hysterothylacium aduncum in the Baltic Sea.

Cod was one of the most important fish species in the Baltic Sea, but its condition is deteriorating for several reasons, including an increasing parasite burden. The aim of this study was to determine the source of infection of Baltic cod with parasites by examination of invertebrates found in situ in the cod stomach. A total of 1681 cod were sampled during four research cruises in the southern Baltic Sea in 2012, 2013 and 2014 and the composition of their diet was analysed. Each prey item from cod stomach was identified to the lowest possible taxonomic level and a parasitological analysis of all invertebrates collected was performed. Crangon crangon, Saduria entomon and Mysis mixta were the most commonly represented invertebrates among food items. Hysterothylacium aduncum was found only in C. crangon. This host­parasite system is reported here for the first time in situ in the stomach of cod from the Baltic Sea, confirming the role of C. crangon in cod infection with H. aduncum.

Anisakid and Raphidascaridid parasites in Trachurus trachurus: infection drivers and possible effects on the host's condition.

This study investigated the distribution of nematode larvae of Anisakidae and Raphidascarididae (genera Anisakis and Hysterothylacium) in Trachurus trachurus (Linnaeus, 1758) in the Ligurian and central-northern Tyrrhenian Seas. The relationship between the number of parasites and the length and weight parameters of the fish was assessed, and the possible effect of the parasites on the condition factor was evaluated. A total of 190 T. trachurus specimens were collected in July 2019. Parasites were found in 70 individuals. A total of 161 visible larvae were collected in the viscera. Morphological analysis revealed the presence of Anisakis spp. in 55 fish and Hysterothylacium spp. in 15 fish, while 5 fish showed coinfection with both genera. The specimens subjected to PCR (n = 67) showed that 85% of the Anisakis larvae analyzed belonged to the species A. pegreffii, while the remaining 15% belonged to hybrids of A. pegreffii-A. simplex (s.s.). A total of 58% (n = 7) of the Hysterothylacium larvae analyzed belonged to the species H. fabri, while 42% belonged to the species H. aduncum. Our results support the hypothesis that infection with these parasites does not affect the condition of the fish host analyzed, and that body size and depth are major drivers in determining infection levels with Anisakid and Raphidascaridid nematodes.

First record of Hysterothylacium fabri (Rudolphi 1819) Deardorff and Overstreet 1980 from Scomber Colias of the South Atlantic waters.

Among several marine pelagic species of the Brazilian coast, Scomber colias Gmelin, 1789 (Perciformes: Scombridae) stands out for having great economic importance, since it is widely used as a food resource and presents moderate vulnerability. Twenty specimens of S. colias were purchased from October 2015 to October 2016 from the coast of Santa Catarina, Brazil. In the present study, we recorded Hysterothylacium fabri (Rudolphi, 1819) (Deardorff and Overstreet, Proc Biol Soc Wash 93(4):1035-1079 1980) from the S. colias intestine using an integrative taxonomy approach, where morphological data are used in combination with partial sequences of the ITS gene, to validate the taxonomic status of the species and establish their relationships with other members of the genus. This species is being recorded for the first time in the South Atlantic and S. colias. The specimens of H. fabri collected in this study parasitizing S. colias presented morphology similar to the other specimens already registered parasitizing other hosts. The distance matrix generated showed that the partial sequences obtained in this study were more similar to sequences of Hysterothylacium sp. collected in China. In phylogenetic analysis, the two detected haplotypes of this study were grouped with H. fabri haplotypes deposited in GenBank in a monophyletic subclade.

Occurrence of larval anisakids in horse mackerel (Trachurus trachurus) caught in Portuguese waters.

The third-stage larvae (L3) of the Anisakidae family are parasitic nematodes with zoonotic impact and are frequently encountered in the organs and musculature of various fish intended for human consumption. Since Anisakis simplex (s.s.) and A. pegreffii are the major aetiological agents of human disease, this study aims to combine the morphological and molecular data on the recovered anisakid larvae to contribute to a simplified morphological distinction of those species and conducted a survey of anisakid larvae infection in horse mackerel (Trachurus trachurus). Here, 116 horse mackerel caught in Portuguese waters were analysed for the presence of L3 of anisakids, and 3148 larvae were collected, of which only 30% were retrieved during visual inspection. As such, visual inspection does not appear to be very effective in anisakid detection. A prevalence of 84.5% of infected fish was found, and the mean intensity and mean abundance were 32.1 and 27.1 parasites per fish, respectively. The morphological and molecular analyses of 196 L3 randomly chosen from the total sample of parasites demonstrated the presence of L3 of mostly Anisakis spp., with only one L3 of Hysterothylacium aduncum. Relative frequencies of 62.9% for A. pegreffii and 37.1% for A. simplex (s.s.) were obtained. The morphometry differences between these two sibling species were evaluated, and the results demonstrated significant differences between the length of the ventriculus and the length of the oesophagus. Precisely, A. simplex (s.s.) has a longer oesophagus and ventriculus than A. pegreffii. As such, these differences may be used to distinguish the two species through morphological analysis.

Morphological and molecular identification of Hysterothylacium larvae (Nematoda: Raphidascarididae) in marine fish from Tunisian Mediterranean coasts.

The taxonomy of Hysterothylacium genus in Mediterranean waters remains incomplete and unresolved. The aim of the current study was to investigate the morphological and molecular identification of selected species of Hysterothylacium larvae in marine fish from the Tunisian Mediterranean coasts. A total of 192 marine fish samples were examined. In total, thirty-seven third-stage larvae of Hysterothylacium were morphologically identified as Hysterothylacium type V. In the present study, representatives of this type from the Mediterranean Sea were genetically characterized for the first time by sequencing the rDNA ITS (ITS1-5.8S-ITS2) regions and mtDNA cox2 gene. This study represents the first report of Hysterothylacium type V from the Mediterranean Sea. We also report Mullus barbatus, M. surmuletus, and Pagellus erythrinus as new hosts for this larval type. Based upon molecular and phylogenetic analyses considering the rDNA ITS regions, the Hysterothylacium type V described here was classified as a new genotype, named Genotype B. The valid genetic data of the described Hysterothylacium type V in the present study can be used to establish the phylogenetic relationships among Hysterothylacium species from the Mediterranean Sea and worldwide for future research.

The hidden companion of non-native fishes in north-east Atlantic waters.

A tropicalization phenomenon of ichthyofauna has been described in the last decades in Galicia (north-eastern Atlantic), with increasing reports of tropical and subtropical fishes appearing northward this distribution range. A search for parasites was carried out in the digestive tract of two specimens first captured in Galician waters: the prickly puffer Ephippion guttifer (Tetraodontidae) and the African stripped grunt Parapristipoma octolineatum (Haemulidae). Examination of E. guttifer showed high intensity of nematodes, from three different genera: Cucullanus (Cucullanidae), Hysterothylacium (Raphidascaridae) and Anisakis (Anisakidae), with demonstrated pathogenicity to humans. Molecular identification allowed the identification of Anisakis pegreffii, already described in the area, and first reports for European waters of Cucullanus dodsworthi, Hysterothylacium reliquens and a new Hysterothylacium sp. P. octolineatum showed a far lower level of parasitization, with two Hysterothylacium larvae, genetically identified as Hysterothylacium deardorffoverstreetorum, also its first report in the eastern Atlantic. Thus, possible ecological impact of the occurrence of two non-native individual fishes in a new area could be remarkably higher if we see this issue through the lens of the parasitological perspective, as far as only two individual fish can harbour more of one hundred nematode parasites belonging to different species, most of them also new species for that area.

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.

Molecular Identification of Anisakis Larvae Extracted by Gastrointestinal Endoscopy from Health Check-up Patients in Korea.

Anisakiasis is a zoonotic disease induced by anisakid nematodes, and endoscopic inspection is used for a diagnosis or remedy for it. Anisakis simplex, Anisakis physeteris, and Pseudoterranova decipiens had been reported to be the major species causing human infections, particularly, in Japan. However, in Korea, recent studies strongly suggested that Anisakis pegreffii is the major species of human infections. To support this suggestion, we collected anisakid larvae (n=20) from 20 human patients who were undergone gastrointestinal endoscopy at a health check-up center in Korea, and molecular identification was performed on the larvae using PCR-RFLP analysis and gene sequencing of rDNA ITS regions and mtDNA cox2. In addition, anisakid larvae (n=53) collected from the sea eel (Astroconger myriaster) were also examined for comparison with those extracted from humans. The results showed that all human samples (100%) were identified as A. pegreffii, whereas 90.7% of the samples from the sea eel were A. pegreffii with the remaining 9.3% being Hysterothylacium aduncum. Our study confirmed that A. pegreffii is the predominant species causing human anisakiasis in Korea, and this seems to be due to the predominance of this larval type in the fish (sea eels) popularly consumed by the Korean people. The possibility of human infection with H. aduncum in Korea is also suggested.

Occurrence of Anisakis and Hysterothylacium Nematodes in Atlantic Chub Mackerels from Libyan Coasts.

The occurrence of zoonotic parasitic nematodes in Atlantic chub mackerels (Scomber colias syn. Scomber japonicus) from Libyan waters was investigated, using epizootiological estimations and molecular specific characterization of larvae. Nematodes belonging to Anisakis spp., the main etiological agent of anisakiasis in Mediterranean waters, and to Hysterothylacium spp. so far considered not pathogenic to humans, were detected. Prevalence values were generally high in visceral cavities (over 40 % for both parasites) while were low for Anisakis (around 1 %) and null for Hysterothylacium in muscles. Moreover, the level of infections was associated with seasons, a feature potentially useful to plan fishing captures and to elaborate risk mitigation strategies for anisakiasis. Species molecular identification performed on a subsample described the presence of Hysterothylacium aduncum as the predominant species, along with Anisakis pegreffii and the hybrids (A. pegreffii and A. simplex sensu stricto), thus posing a concrete zoonotic risk following the consumption of such fish species as a raw preparation.

Morphological and genetic characterization of Hysterothylacium Ward & Magath, 1917 (Nematoda: Raphidascarididae) larvae in horse mackerel, blue whiting and anchovy from Spanish Atlantic and Mediterranean waters.

The presence of zoonotic Hysterothylacium larvae in fish from Spanish Atlantic and Mediterranean waters, which can cause economic losses for commercial fisheries, has been reported in several studies; however, little is known about species identity in this region. The aim of this study was to identify at species level the Hysterothylacium morphotypes detected in three commonly consumed fish: horse mackerel (Trachurus trachurus), blue whiting (Micromesistius poutassou) and anchovy (Engraulis encrasicolus). Third- and fourth-stage Hysterothylacium larvae, as well as adults obtained from larval in vitro culture, were morphologically and molecularly identified by ITS1/ITS2 rDNA sequencing. Four Hysterothylacium morphotypes were detected. Genetic analysis showed that morphotypes VIII and IX were different larval stages of Hysterothylacium aduncum, which was supported by cultured adult species identification. Morphotypes III and IV were found to correspond to different developmental stages of another species of Hysterothylacium. As all larval types detected were morphologically indistinguishable from others previously reported yet showed clear genetic differences, they are referred here as new genotypes. This is the first time that ITS-sequence data of various developmental stages of the same species, including adults, have been studied and compared, providing crucial knowledge for future studies on Hysterothylacium identification and biology.

Occurrence of Anisakis and Hysterothylacium larvae in commercial fish from Balearic Sea (Western Mediterranean Sea).

This study investigates the occurrence of anisakids and raphidascarids in commercial fish from Balearic Sea (Western Mediterranean). A total of 335 fish including 19 black anglerfish (Lophius budegassa), 33 white anglerfish (L. piscatorius), 129 European hake (Merluccius merluccius), 30 red mullet (Mullus barbatus), and 124 striped mullet (M. surmuletus) were examined using enzymatic digestion. A total of 948 nematode larvae were isolated (prevalence 52.53%) being the highest prevalence observed in striped mullet. Forty-six larvae were identified using molecular analyses which included PCR and sequencing of the 629-bp fragment of mitochondrial cox2 gene region. Anisakis pegreffii (80.43%), A. physeteris (8.69%), Hysterothylacium fabri (6.52%), and A. simplex (4.35%) were detected based on molecular analyses of larvae. Total nematode prevalence was positively correlated with weight, length, condition factor, and maturity stage of the host and also with fishing ground depth. Statistical differences between total nematode prevalence and geographical sector of capture were observed when fishing hauls were grouped according to the abundance of sperm whales or common bottlenose dolphins. The results also corroborate that fishing water depth may play an important role in anisakid and raphidascarid parasitization.

Rhabdochona angusticaudata sp. n. (Nematoda: Rhabdochonidae) from the Japanese eel Anguilla japonica, and new records of some other nematodes from inland fishes in Japan.

Collections of parasitic nematodes of fishes from some freshwater and brackish-water localities in Honshu, Shikoku and Hokkaido, Japan, carried out in 1996-2009, yielded the following eight species: Hysterothylacium haze (Machida, Takahashi et Masuuchi, 1978) (Anisakidae), Paraquimperia sp., Pingus sinensis Hsü, 1933 (both Quimperiidae), Heliconema anguillae Yamaguti, 1935 (Physalopteridae), Rhabdochona angusticaudata sp. n., Rhabdochona zacconis Yamaguti, 1935 (both Rhabdochonidae), Ascarophis arctica Polyanskiy, 1952 (Cystidicolidae) and Eustrongylides sp. larvae (Dioctophymatidae). The new species, R. angusticaudata sp. n., described from the intestine of the Japanese eel Anguilla japonica Temminck et Schlegel in the Renjoji River, Shikoku, is mainly characterised by the presence of 14-16 anterior prostomal teeth, non-bifurcated deirids with rounded ends, the length of the left spicule (300-384 µm long), the right spicule with a dorsal barb and non-filamented eggs. The species of Paraquimperia Baylis, 1934 and A. arctica are reported from Japan for the first time. Based on light and scanning electron microscopical examinations, some new morphological data on H. haze, H. anguillae, R. zacconis and A. arctica are provided.

Prevalence and molecular characterization of Hysterothylacium species infecting Pandora (Pagellus erythrinus) in the Mediterranean Sea of Egypt.

Species of the genus Hysterothylacium are aquatic roundworms (nematodes) belonging to the family Raphidascarididae. Some species in this family are known to be associated with zoonotic diseases in humans after they consume their parasitic larvae in raw or undercooked fish. The aim of this research was to report the prevalence, morphology, and molecular characteristics of Hysterothylacium species in Pagellus erythrinus. A total of Two hundred fish were purchased from the fish market in Damanhour, Beheira Province, between December 2021 and November 2022 and subjected to examination. For molecular characterization, the internal transcribed spacer (ITS) region of nuclear ribosomal DNA and the mitochondrial cytochrome oxidase subunit 2 (COX-2) gene were used. Hysterothylacium species were morphologically described and identified from the intestine of Pagellus erythrinus in Beheira Province, Egypt. The PCR amplified 1087 bp and 629 bp of the target sequences of the ITS region and COX-2 gene, respectively. Sequence analysis revealed the Hysterothylacium thalassini species. The identified species provided novel biological data for the Hysterothylacium nematode in Pagellus erythrinus. The prevalence of Hysterothylacium species recovered from the intestine was 55%. The highest prevalence of 72% has been reported in summer compared to the lowest prevalence of 38% in the winter. Females had a higher prevalence of 61.8% than males, with 44.2%. The first detection, prevalence, and molecular characterization of H. thalassini in Pagellus erythrinus from Beheira Province, Egypt, was presented in this study.

Fish feed composition by high-throughput sequencing analysis: Parasite risk assessment.

The use of wild small fish species as feed for aquaculture has clearly an economic incentive by speeding the growth of farmed species. Since feed ingredients are sourced from wild fisheries the farmed species could contain natural contaminants which may introduce food safety concerns. In this study, we used High-Throughput Sequencing (HTS) to explore the whole DNA profile of ten dry commercial feeds commonly used by Spanish fish farming companies. The feeds were mainly made of species within the genus Sprattus, Ammodytes and Clupea, and vegetables of the genus Triticum. In the feeds, DNA sequences of parasitic nematodes of fishes (˂1 % total OTUs) were also identified. A taxonomic assignment of query sequences, using a phylogeny-based approach, estimation of pairwise nucleotide identities within and between sequence groups and haplotype network analysis, allow assign short query sequences to the species Phocanema krabbei (Anisakidae) and Hysterothylacium aduncum (Rhaphidascarididae). Both species were identified as ingredient in two and six fish feeds, respectively. This result is of highly concern regarding dietetic recommendations to sensitized patients to anisakids, considering the growing evidence on the possible allergenic potential of both genera, and the recent data on the transfer of anisakid heat-resistant allergens from fishmeal to farm and aquaculture animals.

Ascaridoid parasites in European sardine throughout the annual cycle: Variability in parasitic load according to host stock features.

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.

Anisakis infection in anchovies (Engraulis encrasicolus) from Iberian waters, southwestern Europe: Post-mortem larval migration.

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.

An Epidemiological Update on Anisakis Nematode Larvae in Red Mullet (Mullus barbatus) from the Ligurian Sea.

Red mullet (Mullus barbatus) is a commercially relevant fish species, yet epidemiological data on anisakid nematode infestation in M. barbatus are scarce. To fill this gap, we report the occurrence of Anisakis larvae in red mullet in the Ligurian Sea (western Mediterranean). This survey was performed between 2018 and 2020 on fresh specimens of M. barbatus (n = 838) from two commercial fishing areas (Imperia, n = 190; Savona, n = 648) in the Ligurian Sea. Larvae morphologically identified as Anisakis spp. (n = 544) were characterized using PCR-RFLP as Anisakis pegreffii. The overall prevalence of A. pegreffii was 24.46%; the prevalence at each sampling site was 6.32% for Imperia and 29.78% for Savona. Furthermore, 3300 larvae of Hysterothylacium spp. were detected in the visceral organs of fish coinfected with A. pegreffii, showing that coinfection with two parasitic species is not rare. This study provides a timely update on the prevalence of ascaridoid nematodes in red mullet of the Ligurian Sea, an important commercial fishing area in the Mediterranean.

Ascaridoid nematodes infecting commercially important marine fish and squid species from Bangladesh waters in the Bay of Bengal.

Parasitic ascaridoid nematodes occur in a wide range of marine organisms across the globe. Some species of the anisakid family (Ascaridoidea: Anisakidae) can cause gastrointestinal disease in humans (i. e. anisakidosis). Despite their importance as potentially hazardous parasites, the occurrence and infection characteristics of ascaridoids are still poorly known from many host species and geographical areas. This study investigated the diversity and infection levels of ascaridoid parasites in various commercial fish and squid host species off Bangladesh. Fish and squid specimens were visually inspected for nematodes using the UV-press method. Nematodes were assigned to genus level based on morphology and identified by sequence analyses of the entire ITS region and partial 28S rDNA and mtDNA cox2 genes. Third-stage larvae (L3) of Anisakis typica occurred at low prevalence (P = 10% and 8%, respectively) in the viscera of Selar crumenophthalmus and Trichiurus lepturus, while Hysterothylacium amoyense occurred in the viscera of Sardinella fimbriata (P = 1%) and the viscera and muscle of Harpadon nehereus (P = 32%) and T. lepturus (P = 76%). Lappetascaris sp. Type A L3 occurred in the mantle of the squid Uroteuthis duvaucelii (P = 11%). Anisakis and Lappetascaris species, and H. amoyense were firstly identified in the Bay of Bengal. The potentially zoonotic A. typica was only found in fish viscera. Hysterothylacium amoyense and Lappetascaris sp., both generally regarded as non-zoonotic, occurred at low prevalence in the muscle or mantle of fish or squid, respectively. Since consumption of raw or lightly processed seafood seems to be rare in Bangladesh, the risk of acquiring anisakidosis from consuming fishery products from off Bangladesh appears to be low. Due to its reddish appearance, the visual presence of H. amoyense larvae in fish flesh may represent a food quality issue.

Nematode Parasites of the European Pilchard, Sardina pilchardus (Walbaum, 1792): A Genuine Human Hazard?

The European pilchard is one of the most frequently consumed fish species in Mediterranean countries, especially in Italy and Spain, and has been reported as the cause of at least eight human anisakidosis cases in Spain. With the aim to shed light on the potential human parasitosis risk posed by nematode larvae belonging to families Anisakidae or Raphidascarididae, a total of 350 sardines captured in the Atlantic Ocean (175 specimens) and the Mediterranean Sea (175 specimens), acquired in various Spanish nationwide supermarket chains, were helminthologically analyzed. The statistical analysis of some helminth parameters revealed a higher presence of nematodes belonging to the genus Hysterothylacium (prevalence 24.29%; mean abundance of 2.36), usually considered non-parasitic for humans (only three cases reported worldwide), when compared to nematodes of the genus Anisakis (5.71%; 0.16). The human anisakidosis risk after the consumption of raw or undercooked sardines and the role of Hysterothylacium, the most frequent nematode, is discussed, providing information to consumers. To avoid human infection by anisakid larval nematodes, the established preventive measures are confirmed and new ones are proposed, such as the consumption of sardines preferably caught in the Mediterranean and of small-sized specimens available, and the immediate evisceration after fishing.