GEOGRAPHICAL VARIATION OF ENTEROPARASITISM IN THE SOUTHERN SEA LION OTARIA FLAVESCENS (SHAW, 1800) OFF THE COAST OF CHILE AS EVIDENCED WITH COPROLOGICAL TESTS: IMPLICATONS FOR ZOONOTIC RISKS.

The scarce information on the helminth fauna in otariids from the Southeastern Pacific comes mainly from stranded individuals or killed for that purpose. In this study, we compared the abundance and composition of enteroparasitic assemblages of Otaria flavescens using coprological techniques. Three sampling localities from north to south spanning 2,200 km off the Chilean coast were considered (Iquique, Viña del Mar, and Talcahuano). In all, 60 fecal samples were collected, and eggs belonging to 5 taxa were found in 91.6% of the samples. They were the anisakid nematodes Contracaecum and Pseudoterranova, the cestode Adenocephalus (syn. Diphyllobothrium), the trematode Ogmogaster, and the acanthocephalan Corynosoma. Samples from southern Chile (Talcahuano) showed the highest prevalence. Adenocephalus eggs had the highest prevalence and abundance in Iquique and Talcahuano, whereas Ogmogaster was the less prevalent and abundant in all sampling localities. Corynosoma eggs had similar prevalence and abundance among sampling localities, and Pseudoterranova eggs were absent in Iquique and with median prevalence values in Viña del Mar and Talcahuano. Thus, the composition of parasite egg assemblages was different between sampling localities. These differences between sampling localities may help to explain differential records of some zoonotic parasitoses such as pseudoterranovosis and diphyllobothriosis in Peru and Chile, where consumption of raw or marinated fish (ceviche) is common. For example, the lower diversity of parasite egg assemblages in the northern Chilean coast may be due to the absence or lower abundance of first intermediate/paratenic hosts of Pseudoterranova.

PHYLOGEOGRAPHY OF BAYLISASCARIS PROCYONIS (RACCOON ROUNDWORM) IN NORTH AMERICA.

Sequences of the mitochondrial cytochrome c oxidase 1 (COI) gene of 115 Baylisascaris procyonis individuals from 13 U.S. states and 1 Canadian province were obtained from 44 raccoon hosts to assess genetic variation and geographic structure. The maximum genetic distance between individuals was low (1.6%), consistent with a single species. Moderate COI haplotype (h = 0.60) and nucleotide (π = 0.0053) diversity were found overall. Low haplotype diversity was found among samples east of the Mississippi River (h = 0.036), suggesting that historical growth and expansion of raccoon populations in this region could be responsible for high parasite gene flow or a selective sweep of B. procyonis mtDNA. There was low genetic structure (average Φst = 0.07) for samples east of the continental divide, but samples from Colorado showed higher diversity and differentiation from midwestern and eastern samples. There was marked genetic structure between samples from east and west of the continental divide, with no haplotypes shared between these regions. There was no significant isolation by distance among any of these geographic samples. The phylogeographic patterns for B. procyonis are similar to genetic results reported for their raccoon definitive hosts. The phylogeographic divergence of B. procyonis from east and west of the continental divide may involve vicariance resulting from Pleistocene glaciation and associated climate variation.

The mitogenome of Ophidascaris wangi isolated from snakes in China.

Different species of the genus Ophidascaris (Baylis, 1921; Nematoda: Ascaridida, Ascaridoidea) are intestinal parasites of various snake species. More than 30 Ophidascaris species have been reported worldwide; however, few molecular genetic studies have been conducted on this genus. We sequenced the complete mitogenome of Ophidascaris wangi parasitizing two snake species of the family Colubridae, i.e., Elaphe carinata (Günther, 1864) and Dinodon rufozonatum. The mitogenome sequence of O. wangi was approximately 14,660 base pairs (bp) long and encoded 36 genes, including 12 protein-coding genes (PCGs), 2 ribosomal RNA (rRNA) genes, and 22 transfer RNA genes. Gene arrangement, genome content, and transcription direction were in line with those in Toxascaris leonina (Linstow, 1902; Ascaridida: Ascarididae). Phylogenetics of O. wangi and other ascaridoids were reconstructed based on the concatenated amino acid sequences of 12 PCGs, and on nucleotide sequences of 12 PCGs and two rRNA genes. Phylogenetic analyses were performed using maximum likelihood and Bayesian inference methods, and the results suggested that O. wangi constitutes a sister clade of Ascaris, Parascaris, Baylisascaris, and Toxascaris within the family Ascarididae, which is a sister clade of Toxocaridae. The mitogenome sequence of O. wangi obtained from the present study will be useful for future identification of the nematode worms in the genus Ophidascaris and will increase the understanding of population genetics, molecular epidemiology, and phylogenetics of ascaridoid nematodes in snakes.

First Report of Resistance to Ivermectin in Parascaris univalens in Iceland.

Horses in Iceland have been isolated for more than 1,000 yr but still harbor a similar range of gastrointestinal parasites as do horses across the world. The long isolation of the horses and their parasites presumably means that no resistance genes have been introduced into the Parascaris spp. population. It is therefore of particular interest to investigate the efficacy of ivermectin on Parascaris spp. infecting Icelandic foals. Potential treatment failure of ivermectin in Iceland will add substantial new information on how resistance can arise independently. This study aimed to determine the efficacy of subcutaneous injection of ivermectin for the treatment of Parascaris spp. infection in foals and to identify the Parascaris species present in the west and north of Iceland. A fecal egg count reduction (FECR) test (FECRT) was performed on 50 foals from 8 farms, including an untreated control group of 6 foals, from September to November 2019. The foals were between 3 and 5 mo of age at the start of the study and had not previously been treated with anthelmintic drugs. Each foal was treated subcutaneously with off-label use of Ivomec® injection 10 mg/ml or Noromectin® 1% at a dose of 0.2 mg/kg. The FECR for each farm was calculated in 2 ways, by the eggCounts package in R and by the Presidente formula (FECRT). Both calculation methods resulted in efficacy levels between 0% and 80.78%, indicating ivermectin resistance on all farms. We also confirmed, by karyotyping, that the species of equine ascarid present in the west and north of Iceland is Parascaris univalens. This study provides evidence for treatment failure of ivermectin against P. univalens infection in foals. Since Icelandic horses have been isolated on the island for more than 1,000 yr, this implies that resistance alleles have developed independently in the Icelandic Parascaris population. The actual clinical impact of ivermectin resistance is unknown but another drug of choice should be considered to treat Parascaris infection in foals in Iceland.

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.

Integrative species delimitation and community structure of nematodes in three species of Australian flathead fishes (Scorpaeniformes: Platycephalidae).

This study aimed to determine the integrative characterisation of nematodes from three species of edible flathead fishes (Scorpaeniformes: Platycephalidae) in New South Wales, Australia, and describe nematode communities within three species of flatheads. Tiger (Platycephalus richardsoni (Castelnau); n = 20) and sand flatheads (Platycephalus bassensis (Cuvier); n = 20), sourced from the Nelson Bay area, and dusky flathead (Platycephalus fuscus (Cuvier); n = 20) from the Manning River, Taree, were examined for the presence of nematodes. The nematodes were initially classified morphologically as 12 different morphotypes belonging to the families Anisakidae (Anisakis types I, II, and III, Contracaecum type II, Terranova types I and II), Raphidascarididae (Hysterothylacium types IV, VI, VIII, and H. zhoushanense larva), and Gnathostomatidae (Echinocephalus sp. larva), Capillariidae (Capillaria sp.), followed by genetic identification through sequencing of the internal transcribed spacer (ITS-1, 5.8S, ITS-2) regions. Phylogenetic analyses revealed the evolutionary relationship between the identified larval specimens in the present study with available GenBank larval and adult nematodes. Sand flathead was 90% infected with nematodes followed by tiger flathead at 85% and dusky flathead at 15%. Nematodes infecting estuarine dusky and oceanic sand and tiger flatheads contrasted markedly. The analysis of similarities (ANOSIM) showed significant differences (p < 0.001) in the composition of taxa within nematode communities between the three species of flatheads (global R = 0.208) with the highest difference being between sand and dusky flatheads (R = 0.308, p < 0.001). The findings of the present study provide a foundation for future investigations of the community composition, life cycles, and distribution of nematode populations in edible fish in Australia and explore and clarify their significance to public health.

Cucullanus carettae Baylis, 1923, in a loggerhead sea turtle (Caretta caretta) from the Adriatic sea: first detection and molecular characterization.

Cucullanus carettae Baylis, 1923 (Nematoda: Cucullanidae) is found worldwide in loggerhead turtles (Caretta caretta). Regarding the Mediterranean, C. carettae has been identified in the Tyrrhenian and the Ionian Sea and a unique description of a Cucullanus sp. specimen in loggerheads from the Adriatic Sea has been reported in the literature so far. In the framework of a bio-monitoring project of the Abruzzo and Molise coasts, a parasitological survey was performed on stranded and by-caught sea turtles, at the Istituto Zooprofilattico of Abruzzo and Molise "G. Caporale." During necropsy, the gastrointestinal system of 72 stranded loggerhead turtles was analyzed for the presence of endoparasites and fecal samples were collected for coprological examination. Adult C. carettae (n = 123) was found in the upper intestine of one loggerhead turtle, associated with chronic lymphoplasmocytic enteritis. Additionally, five stool samples (6.9%) were positive for Cucullanus sp. eggs. Molecular characterization of adult nematodes was carried out to study phylogenetic relationships among the Cucullanus species. To our knowledge, this is the first morphological and molecular identification of C. carettae in loggerhead turtles from the Adriatic Sea. Additional studies on the distribution of this parasite in the Mediterranean are encouraged.

Erection of Euterranova n. gen. and Neoterranova n. gen. (Nematoda, Anisakidae), with the description of E. dentiduplicata n. sp. and new records of two other anisakid nematodes from sharks off New Caledonia.

Helminthological examinations of three species of sharks, Galeocerdo cuvier, Triaenodon obesus (both Carcharhinidae, Carcharhiniformes) and Stegostoma fasciatum (Stegostomatidae, Orectolobiformes) from New Caledonian waters, carried out during 2003-2005, revealed the presence of three species of adult anisakid nematodes referable to Terranova Leiper et Atkinson, 1914. However, this genus can no longer be considered valid, because its type species has been designated a species inquirenda. Therefore, the present nematodes are assigned to two newly established genera, Euterranova n. gen. [type species E. dentiduplicata n. sp.] and Neoterranova n. gen. [type species N. scoliodontis (Baylis, 1931) n. comb.], based mainly on different labial structures. Euterranova dentiduplicata n. sp. from the stomach of S. fasciatum is mainly characterized by the presence of lips with two rows of denticles. Innominate specimens of Euterranova (a female and a third-stage larva) were collected from the digestive tract of T. obesus. Specimens of N. scoliodontis were recorded from G. cuvier. The two named species are described based on light and scanning electron microscopical examinations. Neoterranova scoliodontis has previously been recorded in New Caledonian waters from the same host species. Species previously attributed to Terranova are transferred to Euterranova (5 species), Neoterranova (4 species) or considered species inquirendae (10 species). Since Pseudoterranova Mozgovoy, 1950 was found to be a nomen nudum according to the International Code of Zoological Nomenclature (ICZN), the available name of this genus is Pseudoterranova Mozgovoy, 1953. A key to Porrocaecum-like nematode genera (Porrocaecum, Pseudoterranova, Pulchrascaris, Euterranova, and Neoterranova) is provided.

TITLE: Érection d'Euterranova n. gen. et Neoterranova n. gen. (Nematoda, Anisakidae), avec la description d'E. dentiduplicata n. sp. et de nouveaux signalements de deux autres nématodes Anisakidae de requins au large de la Nouvelle-Calédonie. ABSTRACT: L'examen helminthologique de trois espèces de requins dans les eaux néo-calédoniennes, Galeocerdo cuvier, Triaenodon obesus (tous deux Carcharhinidae, Carcharhiniformes) et Stegostoma fasciatum (Stegostomatidae, Orectolobiformes), réalisé en 2003­2005, a révélé la présence de trois espèces de nématodes Anisakidae adultes qu'on pourrait référer à Terranova Leiper et Atkinson, 1914. Cependant, ce genre ne peut plus être considéré comme valide, car son espèce type a été désignée species inquirenda. Par conséquent, les nématodes décrits ici sont attribués à deux genres nouvellement établis, Euterranova n. gen. [espèce-type E. dentiduplicata n. sp.] et Neoterranova n. gen. [espèce type N. scoliodontis (Baylis, 1931) n. comb.], principalement sur la base de différentes structures labiales. Euterranova dentiduplicata n. sp., de l'estomac de S. fasciatum, se caractérise principalement par la présence de lèvres à deux rangées de denticules. Des spécimens non nommés d'Euterranova (une femelle et une larve de troisième stade) ont été collectés dans le tube digestif de T. obesus. Des spécimens de N. scoliodontis ont été trouvés chez G. cuvier. Les deux espèces nommées sont décrites sur la base d'examens au microscope photonique et électronique à balayage. Neoterranova scoliodontis a déjà été signalé dans les eaux néo-calédoniennes chez la même espèce hôte. Les espèces précédemment attribuées à Terranova sont transférées à Euterranova (5 espèces), Neoterranova (4 espèces) ou considérées comme species inquirendae (10 espèces). Puisque Pseudoterranova Mozgovoy, 1950 s'est avéré être un nomen nudum selon le Code international de nomenclature zoologique (ICZN), le nom disponible de ce genre est Pseudoterranova Mozgovoy, 1953. Une clé des genres de nématodes de type Porrocaecum (Porrocaecum, Pseudoterranova, Pulchrascaris, Euterranova et Neoterranova) est fournie.

Helminth diversity in Pimelodus blochii Valenciennes, 1840 (Osteichthyes: Pimelodidae) in two Amazon Rivers.

Structure of the helminth community and analyses of helminth population parameters of Pimelodus blochii collected in the Xapuri River in comparison with those in the Acre River were evaluated. Eight adult helminth species were found parasitizing P. blochii in the Acre River: the nematodes Orientatractis moraveci, Rondonia rondoni, Philometroides acreanensis, Cucullanus (Cucculanus) pinai pinai, Procamallanus (Spirocamallanus) pimelodus, Rhadochona acuminata, and Brasilnema sp., and the trematode Dadaytrema oxycephala. For Xapuri's fishes, nine helminth species were found: the nematodes O. moraveci, R. rondoni, C. (C.) pinai pinai, Procamallanus (Spirocamallanus) rarus, P. (S.) pimelodus, R. acuminata, Brasilnema sp., and Cystidicolidae gen. sp., and the trematode D. oxycephala. Nematode and Acanthocephala larvae were also reported. Helminth abundance, prevalence, and diversity were influenced by seasonality and locality (river). The helminth parasites from Acre's fishes formed a subset of the helminth community of the Xapuri's. The results indicate an influence of the environmental characteristics of the rivers on the helminth community structure and diversity. This is the first study of the parasite community of P. blochii in the Xapuri River. The paretheses of (Spirocamallanus) and (S.) should not be in italics all along the text and tables.

Infection and molecular identification of ascaridoid nematodes from the important marine food fish Japanese threadfin bream Nemipterus japonicus (Bloch) (Perciformes: Nemipteridae) in China.

The Japanese threadfin bream N. japonicus (Bloch) (Perciformes: Nemipteridae) is an important marine food fish in Asia. However, our present knowledge of the occurrence of its nematode parasites is still limited. In the present study, the species composition and infection rate of ascaridoid nematodes in N. japonicus from the South China Sea, were studied for the first time. Five ascaridoid species, namely Anisakis typica (L3), Hysterothylacium amoyense (L3), Hysterothylacium sp. IV-A (L3), adult of H. thalassini and Raphidascaris lophii (L3), were identified using integrative taxonomy. Hysterothylacium amoyense was the most prevalent species (prevalence 47.2%, mean intensity 14.9 ± 17.1). Hysterothylacium thalassini and R. lophii were reported in the Japanese threadfin bream for the first time. Two different genotypes of A. typica (overall prevalence of 3.4%; mean intensity 1.7 ± 0.9) were found in the South China Sea for the first time. The unique restriction polymorphism patterns of three species of Hysterothylacium are provided for rapid diagnosis. Our present results indicate that RFLP analysis of ITS region, using the restriction enzymes HhaI and RsaI, represents a simple and practical method for large-scale surveys of Hysterothylacium for seafood industry.

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.

New insights into the coexistence of Contracaecum rudolphii A and Contracaecum rudolphii B (Nematoda: Anisakidae) in Phalacrocorax carbo sinensis from Sardinia: genetic variability and phylogenetic analysis.

Contracaecum sp. nematodes are important parasites of fish eating birds that can cause animal health problems. In the present study, specimens of Contracaecum rudolphii sensu lato, from the great cormorant Phalacrocorax carbo sinensis from Sardinia, were characterized based on morphological and molecular data. The morphological analysis allowed to identify all the fourth stage larvae (n = 1918) as Contracaecum sp., and adults, male (n = 5845) and female (n = 8312), as C. rudolphii sensu lato. Population genetics and phylogenetic relationships were inferred based on mitochondrial and nuclear markers. Multiple sequence alignment of the ribosomal internal transcribed spacer showed the coexistence of C. rudolphii A (n = 157), C. rudolphii B (n = 22) and a rare heterozygote of these species. Moreover, mitochondrial markers, namely NADH dehydrogenase subunits I (nad1), cytochrome c oxidase subunit (cox1 and cox2) and small subunit of rRNA (rrnS), showed that the studied C. rudolphii A populations had undergone bottleneck, or founder effect event, subsequent to a rapid population growth and expansion. The observed heterozygote is with a mitochondrial pattern of C. rudolphii B. Although, both Contracaecum species showed high genetic diversity, no genetic structure between localities was detected. Phylogenetic reconstructions supported the paraphyly of the avian Contracaecum species including C. ogmorhini (parasite of otariids).

Some adult and larval nematodes from fishes off New Caledonia.

A collection of nematodes from the Muséum National d'Histoire Naturelle (National Museum of Natural History), Paris, France, was studied. The nematodes had previously been collected from a range of marine fish hosts. The aim of this study was to investigate the identity of these nematodes. Detailed body measurements of the nematodes were taken via light microscopy and where possible first and second internal transcribed spacers (ITS-1 and ITS-2) of ribosomal DNA were subjected to PCR, purified and sequenced. Six species and three larval types were found from a range of fish species and included 13 new host records and two new geographical records. Of the taxa found, Hysterothylacium kajikiae is the only species which may have zoonotic potential. The majority of host fish in this study are edible species, commercially and recreationally fished and frequently incorporated into the cuisine of New Caledonia as raw fish dishes. This study will allow better understanding of the diversity, life cycles, distribution and host-parasite relationships in the New Caledonia area.

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.

Anisakis and Hysterothylacium species in Mediterranean and North-East Atlantic fishes commonly consumed in Spain: Epidemiological, molecular and morphometric discriminant analysis.

The consumption of raw fish parasitized with larval ascaridoid nematodes of the family Anisakidae can cause anisakiasis, provoking gastrointestinal and/or allergic symptomatology. The main causative agents in the Anisakis genus are the sibling species Anisakis simplex sensu stricto (s.s.) and A. pegreffii of the A. simplex sensu lato (s.l.) complex. Larvae of A. simplex (s.l.) are frequently detected in fish commonly consumed in Spain, as are larvae of the genus Hysterothylacium of the family Raphidascarididae, associated with allergic reactions but not considered pathogenic. Reported here are the results of an epidemiological survey of ascaridoid larvae in three commonly consumed fish species in Spain, horse mackerel (Trachurus trachurus) (n = 52), blue whiting (Micromesistius poutassou) (n = 93) and anchovy (Engraulis encrasicolus) (n = 69), caught in the North-Eastern Atlantic, West Mediterranean and Adriatic Sea. The larvae found in the dissected fish were identified in the following order of abundance: A. simplex (s.l.) (n = 2003), Hysterothylacium aduncum (n = 422), H. fabri (n = 180) and A. physeteris (n = 15). Binomial regression analysis showed a correlation between A. simplex (s.l.) and Hysterothylacium larvae abundance and the host geographical location, the North-Eastern Atlantic being the area with the highest parasitation. Fish length and weight and Fulton's condition factor were correlated with A. simplex (s.l.) abundance only in horse mackerel. There was a significant presence of A. simplex (s.l.) and H. aduncum larvae in the musculature of North-Eastern Atlantic blue whiting, the most parasitized part being the anteroventral region, followed equally by the anterodorsal and central sections. The ITS rDNA of larvae of the sibling species A. simplex (s.s.) and A. pegreffii was identified by PCR-RFLP, and a binary logistic regression model was developed to study their morphometric differentiation. Anisakis simplex (s.s.) was detected in the North-Eastern Atlantic and A. pegreffii in all the areas studied. The morphometric analysis discriminated between the two species at the third and fourth larval stages (L3 and L4), the latter obtained by in vitro culture in RPMI-1640 medium. Two discriminant functions were obtained for the L3 and L4 larvae, the ventricle being a key parameter for specific differentiation in both stages, providing taxonomical criteria that could be used besides molecular identification. The present study reveals differences in the parasitation of the studied fish, including the distribution of larvae in the musculature, related to the host species and its geographical origin.

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.

Genetic identification and insights into the ecology of Contracaecum rudolphii A and C. rudolphii B (Nematoda: Anisakidae) from cormorants and fish of aquatic ecosystems of Central Italy.

Contracaecum rudolphii (s. l.) is a complex of sibling species of anisakid nematodes having the fish-eating birds belonging to the Family Phalacrocoracidae as final hosts. The great cormorant Phalacrocorax carbo sinensis is parasitized by C. rudolphii A and C. rudolphii B. Adults and L4 specimens of C. rudolphii (s. l.) (N = 3282) were collected in cormorants from brackish and freshwater ecosystems of Central Italy. Third-stage larvae of Contracaecum (N = 882) were obtained from the fish species Dicentrarchus labrax, Anguilla anguilla, Aphanius fasciatus, Atherina boyeri, Leuciscus cephalus, Barbus barbus, and Carassius carassius captured in the same geographical areas of cormorants' standings. Contracaecum rudolphii A and C. rudolphii B were identified by a multilocus genetic approach: allozymes, sequences analysis of the mtDNA cox2, and ITS region of rDNA gene loci. Differential distribution of the two parasite species was observed in different aquatic environments. Contracaecum rudolphii B outnumbered C. rudolphii A in wintering cormorants from freshwater ecosystems; the opposite trend was found in cormorants from brackish water. Analogously, C. rudolphii A larvae were more prevalent in brackish water fish, while C. rudolphii B larvae were found infecting only freshwater fish. The findings seem to confirm that C. rudolphii A and C. rudolphii B would have a life-cycle adapted to brackish and freshwater environments, respectively. A differential feeding behavior of wintering cormorants, the ecology of the infected fish species, and abiotic factors related to early stages of the parasites are supposed to maintain the distinctiveness of the two parasite species' life cycles in the two different aquatic ecosystems.

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.

Integrative taxonomy of Sprentascaris mahnerti and phylogeny of raphidascaridid nematodes (Nematoda: Ascaridoidea: Raphidascarididae) suggest the monophyly of Sprentascaris.

Sprentascaris mahnerti (Nematoda: Raphidascarididae) collected from Loricariichthys labialis (Siluriformes: Loricariidae) in the Pantanal wetlands, State of Mato Grosso do Sul (Brazil), was redescribed using light and scanning electron microscopy (SEM), and genetically characterised along with two other raphidascaridids: Raphidascaroides brasiliensis and Ro. moraveci. Due to the systematic discussion regarding Raphidascaris and Sprentascaris, as well as the poor knowledge about the phylogenetic relationships within Raphidascarididae, phylogenies were reconstructed based on partial sequences of the 18S and 28S nuclear rRNA gene, the nuclear ITS1-5.8S-ITS2 and the cytochrome c oxidase subunit I (cox1) mtDNA. Morphological study of S. mahnerti, confirmed some previously described features, revealed new characteristics and permitted to elucidate some inconsistencies noted in the literature. Morphological and genetic characterisation of S. mahnerti supported its validity. Phylogenetic reconstructions supported the monophyly of Sprentascaris, which has three pairs of interlabial conspicuous cuticular projections as a synapomorphy. The relationships among several lineages of raphidascaridids were unsolved, albeit Goezia and Ichthyascaris formed well-supported monophyletic assemblages, in which the first included species with no relations regarding the habitat of hosts and the geographic origin. The present findings represent one more step towards the understanding of the interrelationships of raphidascaridid nematodes. In this sense, Sprentascaris should be considered valid as an independent lineage from Raphidascaris.