Morphological identification of Skrjabinisakis Mozgovoi, 1951 (Nematoda: Anisakidae) in Kogia sima (Cetacea: Kogiidae) from Brazilian waters.

New morphological, morphometric and scanning electron microscopy data of a nematode of the family Anisakidae, recovered from a specimen of Kogia sima, a cetacean that died off the northern coast of Brazil, are presented in this paper. Morphological features such as the violin-shaped ventricle and short and equal spicules, as well as the distribution of post-cloacal papillae and specificity for the definitive host (Kogiidae cetaceans) demonstrate similarity to Skrjabinisakis paggiae. This research records Kogia sima and S. paggiae on the estuarine coast of Pará, northern Brazil.

Intrinsic properties of anisakid nematode larvae as a potential tool for the detection in fish products.

Anisakid nematode larvae (NL) in fish products comprise a risk to human health and, if visible, lead to the rejection of these products by consumers. Therefore, great efforts are being made for the identification of these anisakid larvae to estimate the potential consumer health risk as well as to develop effective detection methods in order to prevent the introduction of heavily infected fish products into the market. The tasks of national reference laboratories include the improvement of detection methods and to promote their further development. As a prerequisite for improved detection, it is important to understand the structural properties of anisakid NL and compounds produced during host-parasite interactions. This review provides an overview of the intrinsic properties of anisakid NL and reports the latest detection methods in published literature. First, in order to define the potentially interesting intrinsic properties of anisakid nematodes for their detection, anatomy and compounds involved in host-parasite interactions are summarised. These can be used for various detection approaches, such as in the medical field or for allergen detection in fish products. In addition, fluorescence characteristics and their use as both established and promising candidates for detection methods, especially in the field of optical sensing technologies, are presented. Finally, different detection and identification methods applied by the fish processing industries and by control laboratories are listed. The review intends to highlight trends and provide suggestions for the development of improved detection and identification methods of anisakid NL in fish products.

A new species of Contracaecum (Nematoda: Anisakidae) found parasitizing Nannopterum brasilianus (Suliformes: Phalacrocoracidae) and Hoplias argentinensis (Characiformes: Erythrinidae) in South America: morphological and molecular characterization of larval and adult stages.

Nematode species of the genus Contracaecum Railliet & Henry, 1912 have been reported around the world in many species of fish-eating birds and seals. Here, Contracaecum jorgei n. sp. is morphologically described using light and scanning electron microscopy for adults and fourth-stage larvae (L4) found in the bird Nannopterum brasilianus and third-stage larvae (L3) found in the freshwater fish Hoplias argentinensis, both from the province of Córdoba, Argentina. Additionally, sequences of cytochrome c oxidase subunit II were obtained from these specimens and molecular phylogenetic analysis was used to determine its relationships within the genus. The present species is distinguished from other species by the number and disposition of cephalic papillae; shape and size of the interlabia; length of the spicules; and number and arrangement of papillae in the posterior end of the male. Furthermore, in the molecular analyses, sequences obtained from adult L4 and L3 specimens of C. jorgei n. sp. were similar and grouped, forming an independent lineage, thus confirming it as a distinct species. Thus, morphological characteristics associated with molecular data support the proposal of a new species.

First report of Anisakis sp. (Nematoda: Anisakidae) parasitizing Muscovy duck in Marajó Island, state of Pará, Brazil.

Among the avian used for human consumption, the Muscovy duck is well adapted to various climatic conditions and its breeding is widespread due to its easy handling, and its meat is widely consumed and appreciated, especially in the cuisine of northern Brazil. The present study aimed to report and identify taxonomically the nematodes found in the esophagus of Muscovy ducks reared and marketed in the municipality of Soure, Marajó Island, state of Pará, Brazil, and discuss its zoonotic potential to human. The samples consisted of 30 specimens of Cairina moschata domestica analyzed. A total of 258 nematodes were recovered, which were strongly fixed in the esophageal mucosa of hosts. The morphological and morphometric characteristics were compatible with Anisakis third-stage larvae.

First case of human anisakiasis in Poland.

Consumption of raw or inadequately processed marine fish may result in anisakidosis - a zoonotic disease caused by larvae of the parasitic nematodes of the family Anisakidae (anisakiasis when caused by members of the genus Anisakis (Nematoda: Anisakidae)), commonly found in a variety of marine fish species all over the world. Most cases of anisakidosis have been detected in the residents of Japan and South Korea, which results from the tradition of eating raw and semi-raw fish dishes. However, the disease is now increasingly often diagnosed in other parts of the world, including Europe (mainly in Spain and Italy). In Poland, no cases of human infection with anisakid nematodes have been detected so far. In this study, we report the first case of gastric anisakiasis in Poland, in a 59-year-old female patient, after eating raw Atlantic salmon (Salmo salar). The parasite was identified as the third-stage larva of Anisakis simplex sensu stricto on the basis of morphology and molecular analysis. The larva was still alive and causing pain until it was removed, which occurred more than 5 weeks after infection. The described case prove that anisakiasis should be considered as a potential cause of gastrointestinal tract ailments following the consumption of seafood in countries where no cases of this zoonosis have been reported to date.

First report of Anisakis sp. (Nematoda: Anisakidae) parasitizing Muscovy duck in Marajó Island, state of Pará, Brazil / Primeiro relato de Anisakis sp. (Nematoda: Anisakidae) parasitando pato doméstico na Ilha de Marajó, estado do Pará, Brasil

Abstract Among the avian used for human consumption, the Muscovy duck is well adapted to various climatic conditions and its breeding is widespread due to its easy handling, and its meat is widely consumed and appreciated, especially in the cuisine of northern Brazil. The present study aimed to report and identify taxonomically the nematodes found in the esophagus of Muscovy ducks reared and marketed in the municipality of Soure, Marajó Island, state of Pará, Brazil, and discuss its zoonotic potential to human. The samples consisted of 30 specimens of Cairina moschata domestica analyzed. A total of 258 nematodes were recovered, which were strongly fixed in the esophageal mucosa of hosts. The morphological and morphometric characteristics were compatible with Anisakis third-stage larvae.

Resumo Entre as aves utilizadas para consumo humano, o pato doméstico está bem adaptado às várias condições climáticas. Sua criação é muito difundida devido ao seu fácil manejo. E sua carne é amplamente consumida e apreciada, principalmente na culinária do norte do Brasil. O presente estudo teve como objetivo relatar e identificar taxonomicamente os nematódeos encontrados no esôfago de patos domésticos, criados e comercializados no município de Soure, Ilha de Marajó, estado do Pará, Brasil. E também discutir o seu potencial zoonótico para o homem. As amostras consistiram em 30 espécimes de Cairina moschata domestica analisados. Um total de 258 nematódeos foram recuperados, fortemente fixados na mucosa esofágica dos hospedeiros. As características morfológicas e morfométricas foram compatíveis com as larvas de terceiro estágio de Anisakis.

First evidence of the presence of Anisakis simplex in Crangon crangon and Contracaecum osculatum in Gammarus sp. by in situ examination of the stomach contents of cod (Gadus morhua) from the southern Baltic Sea.

Cod (Gadus morhua), an important fish species in the Baltic Sea, is the paratenic host for many parasite species, including the zoonotic nematodes, Anisakis sp. and Contracaecum osculatum. We aimed to identify which invertebrate species (found in situ in the fish stomach) are responsible for infection of cod with zoonotic nematodes. We found that Crangon crangon and Gammarus sp., both invertebrate prey species of cod, were infected with Anisakis simplex and C. osculatum, respectively. These host-parasite systems are reported here for the first time, implicating C. crangon and Gammarus sp. as sources of infection of Baltic cod with zoonotic nematodes.

Genetic and morphological characterization of Mawsonascaris vulvolacinata n. sp. (Nematoda: Anisakidae) and associated histopathology in a wild caught cowtail stingray, Pastinachus ater.

There are limited reports of infectious agents affecting Australian cowtail stingrays. In the present study, a new species of ascaridoid nematode belonging to the genus Mawsonascaris is described. The most distinct characteristic features were observed in females (the presence of a polar spine in the eggs and a flap-like projection in the vulval area). An identification key for Mawsonascaris spp. is provided. Additionally, internal transcribed spacers (ITS) sequences were obtained for the new species. Alignment of the ITS sequence of the specimens in the present study with those deposited in GenBank showed that there exists no other highly similar sequence. Phylogenetic analyses resulted in a distinct grouping of our specimens supporting morphological distinction from previously described Mawsonascaris spp. Histology was used to investigate the pathology caused by the infection. Necrosis, inflammation and fibrosis were evident at the border of the nodules formed by parasite. A large number of parasites were present in muscularis mucosae and submucosa but not in the muscularis of the stomach. The parasites were associated with an increased inflammatory response, which was also found in the muscularis mucosae and submucosa. Similar pathology has been described in elasmobranchs infected by cestodes, although with more severe lesions.

Molecular Identification of Hysterothylacium spp. In Fishes From the Southern Mediterranean Sea (Southern Italy).

Samples obtained from 11 teleost fish species collected in waters off of the Mediterranean coasts of Sicily, Italy, were examined for the presence of Hysterothylacium spp. larvae. In total, 3,017 fish samples were examined, and the larvae recovered were used in subsequent phylogenetic studies. Fifty-eight raphidascaridid parasitic nematodes were found in the examined fish, with prevalence values ranging from 0.2% in anchovies to 60% in forkbeard samples. Twenty-seven parasites were identified as Hysterothylacium fabri and Hysterothylacium aduncum by sequencing the following regions: the nuclear ribosomal internal transcribed spacer ( ITS) and the mitochondrial cytochrome c oxidase subunit II ( COXII). Two other larvae collected from Lophius piscatorius had very low identity scores, with ITS and COXII sequences reported in GenBank and high genetic distances (AY603539). Pairwise comparisons between the ITS region of the H. fabri isolated from fishes from Sicilian coastal waters and those isolated from the Mediterranean Sea, Turkey (KC852206), revealed genetic differences ranging from 0.015 to 0.018. Our H. aduncum samples had very low genetic differences to H. aduncum from the Adriatic Sea (KP979763, 0.00-0.003). The concatenated phylogenetic examination of the ITS- COXII sequences by using maximum likelihood analyses indicated 3 distinct clades supported by high bootstrap values. Further molecular identification and detailed morphological analyses are needed to clarify these results and confirm the diversity and relationships within Hysterothylacium.

Occurrence of Anisakis (Nematoda: Anisakidae) larvae in unusual hosts in Southern hemisphere.

Nematodes belonging to the genus Anisakis are important parasites due to their abundance in seafood and health impacts on humans. In the present study Anisakis larvae were found in a number of uncommon hosts including the Grey petrel, Procellaria cinerea, the Little penguin, Eudyptula minor, Blue-lipped sea krait, Laticauda laticaudata and Spinner shark, Carcharhinus brevipinna. Morphological examination showed nematodes in these animals are Anisakis larval type I. Genetic characterisation suggested that the larva from one Grey petrel was Anisakis berlandi, whereas the other larvae from the second Grey petrel and from the little penguin were Anisakis pegreffii. A number of larvae found in Blue-lipped sea krait and Spinner shark were identified as Anisakis typica. This is the first report of infective stage of Anisakis larvae parasitising hosts other than teleost fish. Understanding of the extent of infection and the pathogenicity of anisakid nematodes in hosts found in the present study is important in the conservation studies and management plans of these hosts.

Integrative taxonomy of Anisakidae and Raphidascarididae (Nematoda) in Paralichthys patagonicus and Xystreurys rasile (Pisces: Teleostei) from Brazil.

Thirty-six Paralichthys patagonicus and 30 Xystreurys rasile were collected in the state of Rio de Janeiro, Brazil to investigate the presence of anisakid and raphidascaridid nematodes. Anisakis typica, Terranova sp., Contracaecum sp., Hysterothylacium deardorffoverstreetorum, and Raphidascaris sp. were identified using integrative taxonomy of morphological and genetic data. Morphological and morphometric analysis was conducted using bright field microscopy with scanning electron microscopy for topographic characterization of the cuticular surface. Phylogenetic analysis, using ITS and cox2 molecular targets, clearly demonstrated the species identification of A. typica and H. deardorffoverstreetorum and the high diversity of H. deardorffoverstreetorum. This is the first report of A. typica, H. deardorffoverstreetorum, and Raphidascaris sp. parasitizing P. patagonicus and X. rasile.

Anisakiasis Causing Acute Dysentery in Malaysia.

Human anisakiasis is a zoonosis acquired by eating raw or undercooked infected seafood. Herein, we report a case of acute dysentery caused by anisakiasis in a 64-year-old man in Malaysia. A colonoscopy was performed and a nematode larva was found penetrating the mucosa of the ascending colon. Bleeding was observed at the site of penetration. Y-shaped lateral epidermal cords were seen from the cross section of the worm, which is a prominent feature of Anisakis larva. Molecular analysis using polymerase chain reaction of cytochrome oxidase 2 (cox2) gene confirmed the specimen to be larva of Anisakis simplex.

Occurrence of Hysterothylacium and Anisakis nematodes (Ascaridida: Ascaridoidea) in the tanaka's snailfish Liparis tanakae (Gilbert & Burke) (Scorpaeniformes: Liparidae).

The tanaka's snailfish Liparis tanakae (Gilbert & Burke) (Scorpaeniformes: Liparidae) is an economically important marine fish species in China. However, the helminth parasites of this fish are still poorly known. During a helminthological survey of Chinese marine fishes from 2011 to 2012, we revealed that L. tanakae was heavily infected with third-stage larvae and adults of ascaridoid nematodes (total prevalence 100% and mean intensity 82.3 nematodes per fish). Four species of third-stage larvae Hysterothylacium liparis Li, Xu & Zhang, 2007, H. aduncum (Rudolphi, 1802), Hysterothylacium fabri (Rudolphi, 1819), and Anisakis pegreffii (Campana-Rouget & Biocca, 1955) and a single species of adults H. liparis were differentiated and identified by morphological and molecular methods. The detailed morphology of the four species of third-stage larvae was also studied using light microscopy and scanning electron microscopy. The morphological and molecular characterization of the third-stage larvae of H. liparis was reported. Liparis tanakae represents a new host record for A. pegreffii and H. fabri. In addition, a new name, Hysterothylacium zhoushanense nom. nov. was also given to Hysterothylacium zhoushanensis Li, Liu & Zhang, 2012 to make the latinized specific epithet agree with this neuter generic name.

Genetic and morphological approaches distinguish the three sibling species of the Anisakis simplex species complex, with a species designation as Anisakis berlandi n. sp. for A. simplex sp. C (Nematoda: Anisakidae).

Numerous specimens of the 3 sibling species of the Anisakis simplex species complex (A. pegreffii, A. simplex (senso stricto)), and A. simplex sp. C) recovered from cetacean species stranded within the known geographical ranges of these nematodes were studied morphologically and genetically. The genetic characterization was performed on diagnostic allozymes and sequences analysis of nuclear (internal transcribed spacer [ITS] of ribosomal [r]DNA) and mitochondrial (mitochondrial [mt]DNA cox2 and rrnS) genes. These markers showed (1) the occurrence of sympatry of the 2 sibling species A. pegreffii and A. simplex sp. C in the same individual host, the pilot whale, Globicephala melas Traill, from New Zealand waters; (2) the identification of specimens of A. pegreffii in the striped dolphin, Stenella coeruleoalba (Meyen), from the Mediterranean Sea; and (3) the presence of A. simplex (s.s.) in the pilot whale and the minke whale, Balaenoptera acutorostrata Lacépède, from the northeastern Atlantic waters. No F1 hybrids were detected among the 3 species using the nuclear markers. The phylogenetic inference, obtained by maximum parsimony (MP) analysis of separate nuclear (ITS rDNA region), combined mitochondrial (mtDNA cox2 and rrnS) sequences datasets, and by concatenated analysis obtained at both MP and Bayesian inference (BI) of the sequences datasets at the 3 studied genes, resulted in a similar topology. They were congruent in depicting the existence of the 3 species as distinct phylogenetic lineages, and the tree topologies support the finding that A. simplex (s.s.), A. pegreffii, and A. berlandi n. sp. (= A. simplex sp. C) represent a monophyletic group. The morphological and morphometric analyses revealed the presence of morphological features that differed among the 3 biological species. Morphological analysis using principal component analysis, and Procrustes analysis, combining morphological and genetic datasets, showed the specimens clustering into 3 well-defined groups. Nomenclatural designation and formal description are given for A. simplex species C: the name Anisakis berlandi n. sp. is proposed. Key morphological diagnostic traits are as follows between A. berlandi n. sp. and A. simplex (s.s.): ventriculus length, tail shape, tail length/total body length ratio, and left spicule length/total body length ratio; between A. berlandi n. sp. and A. pegreffii: ventriculus length and plectane 1 width/plectane 3 width ratio; and between A. simplex (s.s.) and A. pegreffii: ventriculus length, left and right spicule length/total body length ratios, and tail length/total body length ratio. Ecological data pertaining to the geographical ranges and host distribution of the 3 species are updated.

Morphological and molecular diagnosis of anisakid nematode larvae from cutlassfish (Trichiurus lepturus) off the coast of Rio de Janeiro, Brazil.

Anisakid nematode larvae from Trichiurus lepturus off coast of Rio de Janeiro were studied using light, laser confocal and scanning electron microscopy, in addition to a molecular approach. Mitochondrial cytochrome c-oxidase subunit 2 (mtDNA cox-2), partial 28S (LSU) and internal transcribed spacers (ITS-1, 5.8S, ITS-2) of ribosomal DNA were amplified using the polymerase chain reaction and sequenced to evaluate the phylogenetic relationships between the nematode taxa. The morphological and genetic profiles confirmed that, of the 1,030 larvae collected from the 64 fish examined, 398 were analysed, of which 361 were Hysterothylacium sp. and 37 were Anisakis typica. Larvae of Hysterothylacium sp. were not identified to the species level due to the absence of similar sequences for adult parasites; however, the ITS sequence clustered in the phylogenetic tree with sequences of H. deardorffoverstreetorum, whereas an mtDNA cox-2 and LSU concatenated phylogenetic analysis demonstrated the presence of two clades, both of them under the same name as the larval H. deardorffoverstreetorum. Data on the occurrence of parasites during the winter and summer months were compared using the t-test. The greatest prevalence and intensity of infection were recorded for larval Hysterothylacium, with a prevalence of 51.56% and an intensity of up to 55 parasites per fish. The larval Anisakis exhibit a higher abundance and intensity of infection in the winter months, and those of Hysterothylacium during the summer. However, the t-test indicated no significant differences between the abundance and intensity of infection recorded during the months of collection for either of these larval nematodes. All sequences generated in this study were deposited in GenBank.

Genetic and morphological characterisation of a new species of the genus Hysterothylacium (Nematoda) from Paralichthys isosceles Jordan, 1890 (Pisces: Teleostei) of the Neotropical Region, state of Rio de Janeiro, Brazil.

Taking into account the difficulties of taxonomic identification of larval anisakid nematodes based on morphological characters, genetic analyses were performed, together with those usually applied, in order to identify anisakid larvae found in the flounder Paralichthys isosceles from the littoral of the state of Rio de Janeiro, Brazil. The analysis of 1,820 larvae revealed a new species, similar to Hysterothylacium MD, Hysterothylacium 2, Hysterothylacium KB and Hysterothylacium sp regarding the absence of the larval tooth, an excretory pore situated below the nerve ring level, and slender lateral alae. Moreover, the new species differs from Hysterothylacium fortalezae and Hysterothylacium reliquens with regard to the number and size of spines present on the tail end and from Hysterothylacium patagonicus by the absence of interlabia. The maximum parsimony and neighbour joining tree topologies based on the 18S ribosomal DNA gene, complete internal transcribed spacer region and cytochrome oxidase 2 (COII) gene demonstrated that the Brazilian larvae belong to Raphidascarididae and represent a unique genetic entity, confirmed as a new Hysterothylacium species. Furthermore, the new species presents COII genetic signatures and shares polymorphisms with Raphidascarididae members. This is the first description of a new anisakid species from Brazil through the integration of morphological and molecular taxonomy data.

Genetic and morphological characterisation of a new species of the genus Hysterothylacium (Nematoda) from Paralichthys isosceles Jordan, 1890 (Pisces: Teleostei) of the Neotropical Region, state of Rio de Janeiro, Brazil

Taking into account the difficulties of taxonomic identification of larval anisakid nematodes based on morphological characters, genetic analyses were performed, together with those usually applied, in order to identify anisakid larvae found in the flounder Paralichthys isosceles from the littoral of the state of Rio de Janeiro, Brazil. The analysis of 1,820 larvae revealed a new species, similar to Hysterothylacium MD, Hysterothylacium 2, Hysterothylacium KB and Hysterothylacium sp regarding the absence of the larval tooth, an excretory pore situated below the nerve ring level, and slender lateral alae. Moreover, the new species differs from Hysterothylacium fortalezae and Hysterothylacium reliquens with regard to the number and size of spines present on the tail end and from Hysterothylacium patagonicus by the absence of interlabia. The maximum parsimony and neighbour joining tree topologies based on the 18S ribosomal DNA gene, complete internal transcribed spacer region and cytochrome oxidase 2 (COII) gene demonstrated that the Brazilian larvae belong to Raphidascarididae and represent a unique genetic entity, confirmed as a new Hysterothylacium species. Furthermore, the new species presents COII genetic signatures and shares polymorphisms with Raphidascarididae members. This is the first description of a new anisakid species from Brazil through the integration of morphological and molecular taxonomy data.