The genus Contracaecum Raillet & Henry (Nematoda: Anisakidae): host and geographical distribution on Neotropical and Antarctic species.

Nematodes of the genus Contracaecum Raillet & Henry, 1912 (Anisakidae, Contracaecinae) have a worldwide distribution. The taxonomy of the genus Contracaecum is well-known nowadays due to several morphometric studies, scanning electron microscopy, and molecular biology. The aim of this work was to review, clarify, and summarize the valid species of the genus Contracaecum parasitizing piscivorous sea birds and mammals from both the Neotropical and Antarctic regions reviewing all scientific available papers and electronic searching data up to date. A checklist on Neotropical and Antarctic Contracaecum spp. was organized through a revision of scientific papers and original descriptions. The systematic online search and the most updated papers were obtained through SCOPUS, Google Scholar, PubMed, Medline, World Register of Marine Species, etc. We provide information about hosts, Neotropical and Antarctic localities where worms were collected, references, molecular markers, and Genbank accession numbers. Twenty-five Neotropical and Antarctic Contracaecum species have been recorded up to date and checked out as valid ones according to the most updated data. Twenty-one species parasitize exclusively fish-eating birds, two species were reported only on marine mammals, and the other two parasitize both sea birds and mammals. A total of 20 Contracaecum species are exclusively reported for the Neotropical region, three only for Antarctic hosts, and two species were reported parasitizing both Neotropical and Antarctic hosts. Several Contracaecum species (10) have been corroborated by molecular analysis of different genetic markers. After reviewing all morphological descriptions of the Contracaecum species, and despite most of them have been characterized only by morphometric methods, we are convinced that all species listed in this work correspond to good and valid Contracaecum Neotropical and Antarctic species. Present results indicate that more taxonomic and molecular studies are needed to advance the understanding of the distribution and host specificity of the Contracaecum species.

Morphological and molecular characterization of Hysterothylacium spp. parasitizing Pomatomus saltatrix and Pagrus pagrus of the State of São Paulo, Brazil.

Raphidascarid nematodes have been the focus of several studies, mainly due to the zoonotic potential of some species, even though the cases are underreported. Due to the difficulty in identifying their larvae, the use of diagnostic techniques involving morphological and molecular analyses has grown in the last 20 years. The present study had as objective the morphological and molecular characterization of the L3 larval types of Hysterothylacium collected in Pomatomus saltatrix and Pagrus pagrus from the Brazilian coast, close to the municipality of Santos, State of São Paulo. Twenty specimens of P. saltatrix were necropsied and Hysterothylacium type V (n = 257) and Hysterothylacium type X (n = 5) larvae were found. Five specimens of P. pagrus were necropsied and all were parasitized by Hysterothylacium type V larvae. The analyses showed a genetic proximity relationship between Hysterothylacium types V with other Hysterothylacium V and with H. deardorffoverstreetorum, although this is a species inquirenda. Haplotypes for Hysterothylacium type X found in the present study formed a monophyletic group with other Hysterothylacium X, H. amoyense, and H. zhoushanense. Through this study, new hosts and localities were registered for Hysterothylacium type V and Hysterothylacium type X.

Occurrence and molecular identification of nematodes from blue mackerel Scomber australasicus Cuvier in Australian waters.

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.

Morphological and genetic characterization of Porrocaecum angusticolle (Molin, 1860) (Nematoda: Ascaridomorpha) from the common buzzard Buteo buteo (Linnaeus) (Accipitriformes: Accipitridae) in Czech Republic.

Porrocaecum angusticolle is a nematode species mainly parasitic in the birds of Accipitriformes and Strigiformes. However, some aspects of the morphology of P. angusticolle remain insufficiently known. In the present study, the detailed morphology of P. angusticolle was studied using light and, for the first time, scanning electron microscopy, based on newly collected specimens from the common buzzard Buteo buteo (Linnaeus) (Accipitriformes: Accipitridae) in Czech Republic. Some previously unreported morphological features of taxonomic significance were observed. The nuclear and mitochondrial DNA markers, including partial large ribosomal DNA (28S), complete internal transcribed spacer (ITS-1 + 5.8S + ITS-2), cytochrome c oxidase subunit 1 (cox1) and subunit 2 (cox2) of P. angusticolle were sequenced for molecular identification of this species. There was no intraspecific genetic variation detected in the 28S and ITS regions among different individuals of P. angusticolle, but low level of intraspecific nucleotide divergence was found in the cox1 (0.26-0.78%) and cox2 regions (1.0%). The 28S and cox2 of P. angusticolle were sequenced for the first time. Our molecular evidence supported the validity of both P. angusticolle and P. depressum. The newly obtained genetic data are helpful for further studies of DNA-based taxonomy, population genetics and phylogeny of the genus of Porrocaecum.

The morphological and molecular characterization of Baylisascaris devosi Sprent, 1952 (Ascaridoidea, Nematoda), collected from Pine marten (Martes martes) in Iran.

BACKGROUND: Baylisascaris devosi is an intestinal nematode found in several carnivores including fisher, wolverine, Beech marten, American marten and sable in different parts of the world, but this nematode has not been reported from Pine marten. Therefore, this study aimed to identify Baylisascaris isolated from a Pine marten in Iran using morphological and molecular approaches. METHODS: Specimens of B. devosi were collected from one road-killed Pine marten in northern Iran. Morphological features were evaluated using scanning electron microscopy, energy dispersive x-ray analysis and ion sectioning. The molecular characterization was carried out using partial Cox1, LSU rDNA and ITS-rDNA genes. RESULTS: The nematodes isolated from the Pine marten were confirmed to be B. devosi based on the morphological features and the sequence of ribosomal and mitochondrial loci. X-ray scans (EDAX) were completed on gallium cut structures (papillae, eggs, male spike and mouth denticles) of B. devosi using a dual-beam scanning electron microscope. The male spike and mouth denticles had a high level of hardening elements (Ca, P, S), helping to explain the chemical nature and morphology of the worm. Based on these genetic marker analyses, our sequence had the greatest similarity with Russian B. devosi isolated from sable. CONCLUSIONS: In this study, to our knowledge, the occurrence of B. devosi infection in Pine marten is reported for the first time. Molecular analysis showed that these three genes are suitable molecular markers for identification and inferring phylogenetic relationships of Baylisascaris species. Furthermore, the high divergence of Cox1 between Baylisascaris species indicates that Cox1 could be used for their phylogenetic and taxonomic studies.

Nasal localization of a Pseudoterranova decipiens larva in a Danish patient with suspected allergic rhinitis.

Pseudoterranoviasis is a zoonotic disease caused by nematode larvae of species within the genus Pseudoterranova (seal worm, cod worm). Most infections are gastrointestinal, oesophageal or pharyngeal, but here we report a nasal infection. A 33-year-old patient suffering from rhinitis for 1.5 years recovered a worm larva from the nose. Diagnosis was performed by morphological and molecular characterization, showing the causative agent to be a third-stage larva of Pseudoterranova decipiens (sensu stricto). Various infection routes are discussed.

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.

Description and genetic characterisation of Pulchrascaris australis n. sp. in the scalloped hammerhead shark, Sphyrna lewini (Griffin & Smith) in Australian waters.

Being listed in the IUCN Red List of Threatened Species, knowledge on the biology, health and diseases of the scalloped hammerhead, Sphyrna lewini (Griffith & Smith) is limited; this is especially true for its parasites. In this paper, a new species, Pulchrascaris australis, is morphologically described followed by genetic characterisation based on the sequence of the ITS region. The new species can be easily differentiated from its congeners based on the morphology of the mouthpart, spicules, plectanes, eggs and vulva. Phylogenetic analyses clearly distinguish specimens in the present study from other parasitic nematodes found in the Australasian waters and elsewhere. The sequencing data also suggest that Terranova larval type I found previously in various fish from New Caledonian and Australian (Queensland) waters also belong to Pulchrascaris australis n. sp.

A Case of Angusticaecum holopterum (Rudolphi, 1819) in a Turtle (Testudo graeca)

Seven nematodes collected from a crushed turtle were brought to Selçuk University Faculty of Veterinary Parasitology Department. A few of them were transparented in lactophenol approximately for three weeks. After the parasites were cleared, their head regions were examined microscopically in apical and lateral positions. The posterior ends of the samples were also examined to separate male and females. According to the results of the microscopic examinations, it was found that five of seven parasites were female and remain two were male and parasites were identified as Angusticaecum holopterum (Rudolphi, 1819) considering their morphological characteristics.

First record of Lagochilascaris minor (Nematoda: Ascarididae) in Leopardus geoffroyi (Carnivora: Felidae) in Brazil / Primeiro registro de Lagochilascaris minor (Nematoda: Ascarididae) em Leopardus geoffroyi (Carnivora: Felidae) no Brasil

Abstract Lagochilascariasis is a parasitic disease caused by nematodes of the genus Lagochilascaris. These parasites occur in the neotropical region and their definitive hosts are wild animals, domestic dogs and felids and, accidentally, humans. Here, infection by Lagochilascaris minor is recorded for the first time in a wild felid, in Brazil.

Resumo A lagochilascaríase é uma doença parasitária causada por nematoides do gênero Lagochilascaris. Com distribuição neotropical, este parasito tem como hospedeiros definitivos animais silvestres, canídeos e felídeos domésticos e, acidentalmente, seres humanos. Registra-se a primeira ocorrência de Lagochilascaris minor em um felídeo silvestre no Brasil.

First record of Lagochilascaris minor (Nematoda: Ascarididae) in Leopardus geoffroyi (Carnivora: Felidae) in Brazil.

Lagochilascariasis is a parasitic disease caused by nematodes of the genus Lagochilascaris. These parasites occur in the neotropical region and their definitive hosts are wild animals, domestic dogs and felids and, accidentally, humans. Here, infection by Lagochilascaris minor is recorded for the first time in a wild felid, in Brazil.

Dichelyne (Cucullanellus) bodiani n. sp. and Cucullanus hansoni Olsen, 1952 (Nematoda: Cucullanidae) from fishes off New Caledonia.

Two species of cucullanid nematodes collected from the intestine of marine fishes off New Caledonia were studied with the use of light and scanning electron microscopy (SEM): Dichelyne (Cucullanellus) bodiani n. sp. from Bodianus perditio (Quoy & Gaimard) (Perciformes: Labridae), characterised mainly by the small size of the body (males and gravid females 2.26-3.13 mm and 2.46-3.32 mm long, respectively), the presence of very long spicules (1.53-1.66 mm in length), the remarkably large seminal vesicle and the arrangement of caudal papillae, is the second known species of Dichelyne Jägerskiöld, 1902 parasitising fishes of the Labridae and the second nominal species of this genus recorded from fishes in New Caledonian waters. Cucullanus hansoni Olsen, 1952, originally described from Hawaii, is now, after 67 years, again reported from its type-host, Sufflamen fraenatum (Latreille) (Tetraodontiformes: Balistidae), from off New Caledonia. The SEM examination of C. hansoni, used in this species for the first time, revealed some new morphological details, such as the presence of a median precloacal cuticular elevation or the shape of deirids and distal tips of spicules, as well as the exact location of caudal papillae and phasmids in the male. This is the seventh species of Cucullanus Müller, 1777 recorded from fishes off New Caledonia.

Detection and quantification of Parascaris P-glycoprotein drug transporter expression with a novel mRNA hybridization technique.

Macrocyclic lactone-resistant Parascaris have been reported throughout the world. In part, the drug resistant phenotype is hypothesized to be associated with ATP-binding cassette transporters known as P-glycoproteins. In many systems, P-glycoproteins efflux drugs out of cells thereby precluding drug binding to target receptors. Parascaris may evade macrocyclic lactone-mediated death by effluxing drugs away from target receptors in the nervous system. Alternatively, P-glycoprotein expression in the gut or body wall could prevent penetration of drugs into the body of the parasite altogether. In the present study, we evaluate expression of Peq-pgp-11 and Peq-pgp-16 using a novel multiple nucleic acid hybridization method. This method allowed for visualization of individual mRNA transcripts within fixed tissue sections of Parascaris adults. Our investigation revealed expression of Peq-pgp-11 and Peq-pgp-16 in the intestine, body wall, nerves, lateral cords, and reproductive tissues of male and female parasites. These results suggest that Pgp could efflux drugs locally at the level of parasite neuronal tissue as well as at sites of entry for drugs such as the hypodermis and intestine. The multiple nucleic acid hybridization method could be useful for providing tissue context for gene expression in a variety of nematode parasites.

Redescription of Porrocaecum semiteres (Zeder, 1800) (Nematoda: Ascaridida) from the Song Thrush Turdus philomelos (Passeriformes: Turdidae).

INTRODUCTION: Porrocaecum semiteres (Zeder, 1800) (Ascaridida: Ascaridoidea) occurs frequently in various birds in Europe. However, the knowledge of the morphology of this species remains insufficient. METHODS: Light and scanning electron microscopy were used to study the detailed morphology of adults of P. semiteres based on newly collected specimens from Turdus philomelos in the Czech Republic. RESULTS: Some previously unreported morphological features were revealed, including the presence of a medio-apical notch, 70-80 small conical denticles and one pair of submedial pores on each lip, the absence of caudal alae, the presence of four pairs of postcloacal papillae, and a single, medio-ventral precloacal papilla.

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.

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.

Morphological and molecular characterization of Seuratascaris numidica (Seurat, 1917) (Ascaridida: Ascarididae).

Seuratascaris numidica (Seurat, 1917) is a specialized nematode species parasitizing amphibians only. In the present study, the detailed morphology of this poorly known species was studied using light and scanning electron microscopy based on the newly material collected from Hoplobatrachus chinensis (Osbeck) (Amphibia: Anura) in China. We found that the relative length of intestinal caecum in our male specimens (representing 68.4-71.1% of oesophageal length) is slighter longer than the previously reported data (not over 60.0% of oesophageal length). Our SEM observations also revealed the presence of ca. 64-76 small conical denticles on each lip. In addition, Angusticaecum wuyiensis Wang, 1981, collected from Rana schmackeri Boettger (Amphibia: Anura) from Wuyi Mountain in Fujian Province, China was considered as a new synonym of S. numidica. The ITS and cox1 sequences of S. numidica were also sequenced for the first time and there is no nucleotide variability detected in both regions. The present supplementary morphological and molecular data (especially the ITS and cox1 sequences) obtained herein is extremely important and useful to determine the morphological variability, population genetics and phylogenetic position of S. numidica in the future.

A frequent roundworm Baylisascaris transfuga in overpopulated brown bears (Ursus arctos) in Slovakia: a problem worthy of attention.

The genus Baylisascaris (order Ascaridida) includes numerous relatively host-specific nematodes, which are common in intestines of wild mammals. Some of them may have impact on veterinary and public health, as their larvae have the potential to cause visceral, ocular, and/or neural larva migrans in a wide range of mammals, birds, and humans. Baylisascaris transfuga is a parasite occurring in a range of bear species throughout the world. We present the current data on B. transfuga occurrence in brown bears from a relatively restricted territory of the Polana Protected Landscape Area in Central Slovakia, obtained by traditional methods (faecal examination, morphology). Species affiliation was confirmed by employing molecular markers generating nuclear 28S and mitochondrial cox1 sequences in adult worms. Based on 17 examined samples (15 excrements and two intestines of young bear females), the occurrence of B. transfuga in the surveyed area was assessed as 52.9%. Both bear females were infected with adult and juvenile worms. Due to the high density of bears in the locality, the high infection rate with ascarids, and the huge number of eggs produced by the parasites, it is apparent that the respective environment, including the inhabited areas, might be markedly contaminated by Baylisascaris eggs. The ability of B. transfuga to serve as a zoonotic agent has not been unambiguously proved; however, this attribute should be considered and subjected to further research.

Morphological variability and molecular characterization of Mawsonascaris australis (Johnston & Mawson, 1943) (Nematoda: Ascaridoidea: Acanthocheilidae) from the brown guitarfish Rhinobatos schlegelii Müller & Henle (Rhinopristiformes: Rhinobatidae).

A rare ascaridoid species, Mawsonascaris australis (Johnston & Mawson, 1943), is reported from the Taiwan Strait for the first time, and the brown guitarfish Rhinobatos schlegelii Müller & Henle (Rhinopristiformes: Rhinobatidae) represents a new fish host. The detailed morphology of M. australis was studied using light and scanning electron microscopy (SEM) based on the newly collected specimens. The results revealed some important, but previously unreported or erroneous, morphological features, including the lateral alae starting well posterior to the base of the ventrolateral lips, the presence of one pair of double paracloacal papillae and a single, small precloacal median papilla on the anterior margin of the cloaca. In addition, in order to determine whether the morphometric difference in the relative lengths of the right and left spicules represents intraspecific or interspecific variation, specimens of M. australis were characterized using molecular approaches by sequencing and analysing the ribosomal internal transcribed spacer (ITS) and mitochondrial cox1 genes. The results indicate that this morphometric difference represents intraspecific variability.

Morphometric and molecular descriptions of three new species of Hysterothylacium (Nematoda: Raphidascarididae) from Australian marine fish.

Three new species of Hysterothylacium Ward & Magath, 1917, including H. australe, H. kajikiae and H. brucei, from Australian marine fish are described and illustrated by light microscopy followed by genetic characterization of their first and second internal transcribed spacers (ITS-1 and ITS-2, respectively). This is the first study reporting ITS sequence data for adult Hysterothylacium spp. in Australia, which provides an insight into the identification of some of the Hysterothylacium larval types. Alignment of ITS sequences of these species with Hysterothylacium larval types previously reported in Australia showed that fourth-stage Hysterothylacium larval type XI from Seriola lalandi and third-stage Hysterothylacium larval type X from Sphyraene novae-hollandiae are identical with ITS sequences of H. australe, suggesting that these fish are natural intermediate/paratenic hosts of H. australe.