Redescriptions of Spinitectus acipenseri and S. micracanthus (Nematoda, Cystidicolidae), with notes on the taxonomy of Spinitectus-like nematodes parasitising North American fishes.

Based on light microscopical and scanning electron microscopical (SEM) examinations, two North American species of Spinitectus Fourment, 1884, S. acipenseri Choudhury & Dick, 1992 and S. micracanthus Christian, 1972 (Nematoda, Cystidicolidae) are redescribed from museum voucher specimens (S. acipenseri) and those newly collected from centrarchid and some other fishes in the Upper San Marcos River in Texas and the Santee River in South Carolina, USA. The first use of SEM to study S. acipenseri, a parasite of lake sturgeon Acipenser fulvescens Rafinesque (Acipenseridae) in Canada, made it possible to describe dorsal and ventral lips, amphids and sublabia, and the presence of a dorsal barb on the right spicule, which was confirmed to be the most characteristic feature of this species. The SEM study of S. micracanthus, a parasite mainly of centrarchids, enabled us to correctly determine the location of the excretory pore in relation to rings of cuticular spines in the male, and to describe the exact structure of the tip of the male tail, sublabia, phasmids and the presence of a median ventral protuberance on the male tail. Some taxonomic problems of North American species of Spinitectus are discussed. Filaria serrata Linton, 1901 is considered a junior synonym of S. oviflagellis Fourment, 1884. To date, there are 13 valid species of Spinitectus parasitising fishes in North America. Keys to species of Spinitectus-like nematodes from fishes in North American waters are provided.

Title: Redescriptions de Spinitectus acipenseri et S. micracanthus (Nematoda, Cystidicolidae), et notes sur la taxonomie des nématodes de type Spinitectus parasitant les poissons nord-américains. Abstract: Deux espèces nord-américaines de Spinitectus Fourment, 1884, S. acipenseri Choudhury & Dick, 1992 et S. micracanthus Christian, 1972 (Nematoda, Cystidicolidae) sont redécrites à partir de spécimens de musée et d'autres nouvellement collectés aux USA dans les rivières Upper San Marcos (Texas) and Santee (Caroline du Sud), avec l'aide de la microscopie optique et électronique à balayage (MEB). L'utilisation pour la première fois du MEB pour étudier S. acipenseri, un parasite de l'esturgeon Acipenser fulvescens Rafinesque (Acipenseridae) au Canada, a permis de décrire les lèvres dorsales and ventrales, les amphides et sublabia, ainsi que la barbe dorsale sur le spicule droit que nous confirmons être le critère le plus caractéristique de cette espèce. L'étude au MEB de S. micracanthus, un parasite principalement de centrarchides, nous a permis de déterminer correctement la place du pore excréteur en relation avec les couronnes d'épines cuticulaires chez le mâle, ainsi que de décrire la structure exacte de l'extrémité caudale du mâle, des sublabia, des phasmides et la présence d'une protubérance médiane et ventrale sur la queue du mâle. Des problèmes taxonomiques sur les espèces nord-américaines de Spinitectus sont discutés. Filaria serrata Linton, 1901 est considéré synonyme plus récent de S. oviflagellis Fourment, 1884. À ce jour, il y a 13 espèces valides de Spinitectus parasites de poissons en Amérique du Nord. Des clés des espèces des nématodes de type Spinitectus provenant des poissons des eaux nord-américaines sont fournies.

Position of Polyclithrum within Gyrodactylidae (Monogenoidea): incongruences between morphological and molecular phylogenies.

This study aimed to enhance our understanding in monogenoid evolution by using morphological and molecular data to determine kinship relationships between species and changes in morphological structures over time. We focused on variations in characteristics among the organisms of the family Gyrodactylidae, concentrating on the phylogenetic position of Polyclithrum with other genera in the family. We collected specimens of Polyclithrum from the striped mullet, Mugil cephalus and Swingleus, and Fundulotrema specimens from mummichog Fundulus heteroclitus in estuarine systems of South Carolina, United States. In addition, we analyzed them and other genera (including e.g., Mormyrogyrodactylus, Gyrodactyloides, and Macrogyrodactylus) using both morphological and molecular (18S rDNA) approaches. We performed phylogenetic trees based on Maximum Parsymony, Maximum Likelihood and Bayesian Inference, and constructed a character morphological matrix by Parsimony Reconstruction of Ancestral Character States method. Our results suggest a homoplastic origin with evolutionary convergences in characters, revealing that there is inconsistency between our data and previously published works based solely on morphological structures of the group. The homoplasy scenario found in Gyrodactylidae can be a result of the limited set of putative homologous morphological features. However, differences between the phylogenies based on morphology and those based on molecular data may arise from both databases. While morphology remains essential in understanding the evolution of this group, molecular data, otherwise, provide a less biased source of information for constructing phylogenetic hypotheses. Combining these data facilitates a better comprehension of the homologous status of morphological features and to understand Gyrodactylidae evolutionary history.

Infection of Atlantic tripletail Lobotes surinamensis (Teleostei: Lobotidae) by brain metacercariae Cardiocephaloides medioconiger (Digenea: Strigeidae).

Three juvenile Atlantic tripletail Lobotes surinamensis caught opportunistically in Charleston Harbor (South Carolina, USA) and maintained in captivity for over three months displayed an altered swimming behavior. While no direct causation can be demonstrated herein, fish were infected in their brain by strigeid trematode larvae (metacercariae) of Cardiocephaloides medioconiger, which were identified via ITS2 and 28S ribosomal RNA gene sequencing. Histology showed nonencysted metacercariae within the brain ventricle between the optic tectum and tegmentum, causing distortion of tegmental parenchyma. Aggregates of mononuclear inflammatory cells were in the ventricle adjacent to metacercariae. Metacercarial infection by Cardiocephaloides medioconiger has been reported from the brain and eyes of only two other fish species from the northern US Atlantic coast: the grey mullet Mugil cephalus and silverside Menidia menidia, but this identification is problematic and needs molecular verification. Atlantic tripletail is a new report as a second intermediate host for C. medioconiger and South Carolina is a new locality. Cardiocephaloides species in general have a low host specificity and infection by C. medioconiger could propagate to other fishes and affect neighboring natural ecosystems.

Ontogenetic and spatial variability in parasite communities of white shrimp Penaeus setiferus (Decapoda: Penaeidae).

Understanding the combined effects of multi-parasite infections on their hosts is necessary for documenting parasite impacts and is particularly important for developing effective management strategies for economically important organisms. The white shrimp Penaeus setiferus supports important recreational and commercial fisheries along the southeastern and Gulf coasts of the United States and occupies an important ecological niche in estuarine and offshore habitats throughout these regions. The goal of this study was to identify and assess ontogenetic and spatial variation in white shrimp parasite communities and their relation to shrimp health. We used a series of trawl surveys in tidal creek and open water habitats of an estuary in the southeastern USA to collect and identify parasites of white shrimp using morphological and DNA sequencing techniques. Parasite communities in white shrimp were composed of organisms belonging to 6 classes: Conoidasida (gregarines), Oligohymenophorea (apostome and sessilid ciliates), Microsporea (meiodihaplophasids), Chromadorea (rhabditids), Cestoda (cyclophyllideans, lecanocephalideans and trypanorhynchs) and Trematoda (plagiorchiids). Parasite communities differed significantly among white shrimp life stages and localities. Furthermore, the health condition known as black gill occurred in some shrimp and was significantly related to parasite community structure. Infection metrics for the apostome ciliate Hyalophysa lynni, the trypanorhynch larvae Prochristianella sp. and the rhabditid larvae Hysterothylacium sp. were significantly different between shrimp exhibiting and not exhibiting black gill. These results highlight the importance of understanding parasite communities and the potential interactive effects of multiple parasite infections on shrimp health.

Parasites of Moroccan desert Coptodon guineensis (Pisces, Cichlidae): transition and resilience in a simplified hypersaline ecosystem.

Sebkha Imlili (Atlantic Sahara) is a salt flat with over 160 permanent holes of hypersaline water generated in the Holocene and inhabited by euryhaline organisms that are considered to be relics of the past, including the cichlid fish Coptodon guineensis. We surveyed the fish parasites four times over one year, to i) identify the parasites, and ii) determine possible seasonality in infection patterns. Over 60% of the fish were infected by one to three helminths: an acanthocephalan in the intestine and two digenean metacercariae in the kidney, spleen, liver, muscle, and mesenteries. The acanthocephalan Acanthogyrus (Acanthosentis) cf. tilapiae was identified morphologically and molecularly; only one digenean (the heterophyid Pygidiopsis genata) could be identified molecularly. Both identified parasites were present throughout the sampling periods; the unidentified metacercariae were present only in summer and fall. Mean intensities, but not prevalence of infection by the acanthocephalan, reflected a biannual pattern of transmission. Infection accrued with fish size, possibly due to cannibalism. Because the water holes include only a few invertebrates, the intermediate hosts of these parasites can be inferred to be the gastropod Ecrobia ventrosa for the digeneans and either the copepod Cletocamtpus retrogressus or the ostracod Cyprideis torosa for the acanthocephalan. This ecosystem appears stable and provides a window into the past, as the acanthocephalan likely switched from freshwater tilapia to C. guineensis when the Sebkha formed. However, this is a vulnerable environment where the survival of these parasites depends on interactions maintained among only very few hosts.

Title: Parasites de Coptodon guineensis (Pisces, Cichlidae) du désert marocain : transition et résilience dans un écosystème hypersalin simplifié. Abstract: La Sebkha d'Imlili est une sebkha (étendue désertique sableuse et salée) dans le Sahara Atlantique caractérisée par la présence de plus de 160 poches permanentes d'eau hypersaline qui sont apparues à l'Holocène et qui sont habitées par des organismes considérés comme des reliques du passé, dont un poisson cichlidé, Coptodon guineensis. Nous avons fait l'inventaire des parasites de ce poisson au cours des quatre saisons d'une année pour 1) identifier les parasites et 2) déterminer une éventuelle transmission saisonnière. Plus de 60 % des poissons étaient infestés par un à trois helminthes : un Acanthocéphale dans l'intestin et des métacercaires de deux espèces de Digène dans le rein, la rate, les muscles et le mésentère. L'Acanthocéphale Acanthogyrus (Acanthosentis) cf. tilapiae a été identifié morphologiquement et génétiquement mais seul un des deux Digènes (l'hétérophyidé Pygidiopsis genata) a pu être identifié par séquençage. Ces deux parasites étaient présents à chaque période d'étude, mais la métacercaire non identifiée était présente seulement en été et en automne. L'intensité moyenne de l'infestation par l'Acanthocéphale, mais pas sa prévalence, reflète une transmission biannuelle. L'infestation augmente avec la taille du poisson, peut-être à cause du cannibalisme. L'identité des hôtes intermédiaires de ces parasites peut être avancée parce que cet écosystème est simplifié et inclut seulement quelques invertébrés : pour les Digènes, le Gastéropode Ecrobia ventrosa, et pour l'Acanthocéphale, le Copépode Cletocamtpus retrogressus ou l'Ostracode Cyprideis torosa. Cet écosystème apparait stable et offre une vue sur le passé étant donné que l'Acanthocéphale a sans doute été transféré d'un tilapia d'eau douce quand la sebkha s'est formée. Cependant, c'est un environnement vulnérable où la survie de ces parasites dépend d'interactions entre très peu d'espèces hôtes.

Limited accrual of myxospores of Kudoa inornata (Cnidaria: Myxosporea) in their wild fish hosts, Cynoscion nebulosus (Teleostei: Sciaenidae).

Kudoa inornata is a myxosporean that infects the seatrout Cynoscion nebulosus. Increased prevalence of infection as fish age and absence of inflammation against plasmodia led to the hypothesis that seatrout retain and accumulate myxospores throughout their lives. However, opportunistic observations that wild-caught seatrout cleared infection when maintained in aquaculture conditions and evidence of encapsulated infected necrotic myofibers suggested that fish develop an immunity against this parasite, or that myxospores have a limited life span. To evaluate myxospore clearance and to test putative resistance to re-infection, we examined 44 wild-caught seatrout broodstock maintained in parasite-free water for 2-6 yr. Twenty-five fish served as negative controls (time zero of experiment), and 19 were exposed to water-borne K. inornata infective stages for 18 wk. Over 73% of the exposed fish became infected, compared to ~12% of control fish, indicating that fish were susceptible to re-infection by K. inornata. Whether plasmodia degenerate because K. inornata myxospores have a limited life span or seatrout develop an adaptive immunity against these life stages remains unknown. To test for accumulation of myxospores over time, we compared myxospore densities and intensities between sexes and across ages and sizes of wild seatrout. There was no significant difference in myxospore densities with size, age, or sex. However, intensities increased significantly with increasing fish age and size, indicating accrual of myxospores over time. These results combined with evidence of infection clearance suggest that K. inornata myxospores do not persist but nevertheless accrue in wild seatrout due to continuous contact with infective stages.

Hexabothriidae and Monocotylidae (Monogenoidea) from the gills of neonate hammerhead sharks (Sphyrnidae) Sphyrna gilberti, Sphyrna lewini and their hybrids from the western North Atlantic Ocean.

Neonates of hammerhead sharks (Sphyrnidae), Sphyrna lewini (Griffith and Smith, 1834), the sympatric cryptic species, Sphyrna gilberti Quattro et al., 2013, and their hybrids were captured in the western North Atlantic, along the coast of South Carolina, USA, between 2018 and 2019 and examined for gill monogenoids. Parasites were identified and redescribed from the gills of 79 neonates, and DNA sequences from partial fragments of the nuclear 28S ribosomal RNA (rDNA) and cytochrome c oxidase I mitochondrial DNA (COI) genes were generated to confirm species identifications. Three species of monogenoids from Hexabothriidae Price, 1942 and Monocotylidae Taschenberg, 1879 were determined and redescribed. Two species of Hexabothriidae, Erpocotyle microstoma (Brooks, 1934) and Erpocotyle sphyrnae (MacCallum, 1931), infecting both species of Sphyrna and hybrids; and 1 species of Monocotylidae, Loimosina wilsoni Manter, 1944, infecting only S. lewini and hybrids. Loimosina wilsoni 28S rDNA sequences matched those of Loimosina sp. from the southern coast of Brazil. Based on limited morphological analysis, Loimosina parawilsoni is likely a junior synonym of L. wilsoni. This is the first taxonomic study of monogenoids infecting S. gilberti and hybrids of S. gilberti and S. lewini.

Molecular data reshape our understanding of the life cycles of three digeneans (Monorchiidae and Gymnophallidae) infecting the bivalve, Donax variabilis: it's just a facultative host!

The coquina, Donax variabilis, is a known intermediate host of monorchiid and gymnophallid digeneans. Limited morphological criteria for the host and the digeneans' larval stages have caused confusion in records. Herein, identities of coquinas from the United States (US) Atlantic coast were verified molecularly. We demonstrate that the current GenBank sequences for D. variabilis are erroneous, with the US sequence referring to D. fossor. Two cercariae and three metacercariae previously described in the Gulf of Mexico and one new cercaria were identified morphologically and molecularly, with only metacercariae occurring in both hosts. On the Southeast Atlantic coast, D. variabilis' role is limited to being a facultative second intermediate host, and D. fossor, an older species, acts as both first and second intermediate hosts. Sequencing demonstrated 100% similarities between larval stages for each of the three digeneans. Sporocysts, single tail cercariae, and metacercariae in the incurrent siphon had sequences identical to those of monorchiid Lasiotocus trachinoti, for which we provide the complete life cycle. Adults are not known for the other two digeneans, and sequences from their larval stages were not identical to any in GenBank. Large sporocysts, cercariae (Cercaria choanura), and metacercariae in the coquinas' foot were identified as Lasiotocus choanura (Hopkins, 1958) n. comb. Small sporocysts, furcocercous cercariae, and metacercariae in the mantle were identified as gymnophallid Parvatrema cf. donacis. We clarify records wherein authors recognized the three digenean species but confused their life stages, and probably the hosts, as D. variabilis is sympatric with cryptic D. texasianus in the Gulf of Mexico.

TITLE: Les données moléculaires remodèlent notre compréhension des cycles de vie de trois Digènes (Monorchiidae et Gymnophallidae) parasites du bivalve Donax variabilis : ce n'est qu'un hôte facultatif ! ABSTRACT: La telline, Donax variabilis, est un hôte intermédiaire connu de digènes Monorchiidae et Gymnophallidae. Le nombre limité de critères morphologiques pour identifier les hôtes et les stades larvaires des parasites sont à la base de confusion dans la littérature. Dans cette étude nous avons identifié par séquençage les tellines collectées sur la côte Atlantique des États-Unis. Nous démontrons que les séquences pour D. variabilis dans GenBank sont incorrectes et que celle du spécimen américain est en fait celle de D. fossor. Deux cercaires et trois métacercaires décrites au préalable dans le Golfe du Mexique ainsi qu'une nouvelle cercaire ont été identifiées morphologiquement et par séquençage. Les métacercaires seules infectent les deux hôtes. Sur la côte sud-est Atlantique, D. variabilis a un rôle limité à seulement celui de second hôte intermédiaire facultatif, et D. fossor, une espèce plus ancienne, est premier et second hôte intermédiaire. Le séquençage a montré 100 % de similarité entre les stades larvaires de chacun des trois digènes. Des sporocystes avec des cercaires à queue simple et les métacercaires dans le siphon inhalant ont des séquences identiques à celles de Lasiotocus trachinoti, pour qui nous donnons le cycle complet. Les adultes des deux autres digènes ne sont pas connus et les séquences des stades larvaires ne sont identiques à aucune dans GenBank. Des sporocystes de grande taille, les cercaires (Cercaria choanura) et les métacercaires enkystées dans le pied des tellines sont identifiées comme étant Lasiotocus choanura (Hopkins, 1958) n. comb. Des petits sporocystes avec des furcocercaires et des métacercaires dans le manteau des tellines sont identifiées comme étant le gymnophallide Parvatrema cf. donacis. Nous clarifions les rapports de la littérature dans lesquels les auteurs ont reconnu les trois espèces de digènes mais ont confondu leurs stades larvaires et probablement les hôtes étant donné que D. variabilis vit en sympatrie avec l'espèce cryptique D. texasianus dans le Golfe du Mexique.

Dynamics of infection and pathology induced by the aporocotylid, Cardicola laruei, in Spotted Seatrout, Cynoscion nebulosus (Sciaenidae).

The sciaenid Spotted Seatrout (Cynoscion nebulosus) are infected by blood flukes (Cardicola spp.). A 2 year survey in estuaries of South Carolina, USA, showed that adult flukes and granulomas occurred throughout the year but their prevalence was highest in summer (61% and 84%, respectively), indicating an unusually high level of infection for wild fish. Granulomas remained after adult flukes could no longer be found. PCR-Restriction Fragment Length Polymorphism (RFLP) of a subsample of specimens allowed identification of Cardicola laruei as the only species infecting these seatrout during the period of study. Mean intensity of infection by flukes was higher in female seatrout, suggesting endocrine and/or immune system involvement. The prevalence of granulomas declined sharply in winter, indicating possible mortality of infected seatrout as this species is known to be cold-sensitive. Granulomas were studied using histology, immunohistochemistry, and transmission electron microscopy. Eggs were encapsulated by an inner core of dark epithelioid cells, and an outer core of large epithelioid cells undergoing epithelialization. Fibrosis was observed around granulomas and some granulomas detached from the surrounding damaged myocardium. Numerous inflammatory cells appeared mobilised around granulomas and pathology could be severe, in some cases showing grossly visible blister-like extrusions scattered in the damaged epicardium. At the gross level, some granulomas possessing eggs with live miracidia were observed at the surface of the epicardium. These findings suggest that granulomas carrying both dead and live eggs can clear the fish heart by host-mediated transport through the myocardium, as is known to occur in related human Schistosoma infections.

Microhabitat preference, body size, and egg allocation in the gill parasite Naobranchia lizae (Copepoda).

The relationships between microhabitat preference, body size, and egg allocation were examined in the copepod Naobranchia lizae, which establishes on the gills of striped mullet Mugil cephalus. A total of 297 individual N. lizae (mean intensity = 5.0 ± 4.8 SD) were recovered from 60 infected hosts collected from the Charleston Harbor Estuarine System, South Carolina USA. For each mullet, we identified 16 microhabitats per gill arch, which yielded 128 microhabitats per host that could potentially be occupied. On average, only 5% of these microhabitats were occupied per host. The distribution pattern of the copepods on the gills revealed that microhabitat preferences occurred both among and within gill arches. For the microhabitats occupied, there was no effect of preference on body size, egg number, or egg size. Similarly, microhabitat sharing, which was more likely to occur at higher infection intensities, was not costly in terms of the copepod body size and egg allocation and there was no detectable trade-off between egg number and egg size. However, results also revealed that about half (48%) of the available microhabitats were never occupied by the copepods. We suggest that the occupancy of these potentially poor quality sites could carry fitness costs not realized in nature since numerous high quality sites are available per host. The findings are consistent with the interpretation that female N. lizae occupy a resource-rich habitat on the gill arches of striped mullet that provides conditions for optimal growth and reproduction.

First record of Piscicapillaria bursata (Nematoda: Capillariidae), a parasite of hammerhead sharks Sphyrna spp., in the western Atlantic Ocean.

Examination of 32 spiral valves from neonate specimens of hammerhead shark Sphyrna spp. (Carcharhiniformes: Sphyrnidae) captured between June and August 2018 off the Atlantic coast of South Carolina, USA, revealed the presence of the capillariid nematode Piscicapillaria bursata (Capillariidae) in the Carolina hammerhead S. gilberti, the scalloped hammerhead S. lewini, and their hybrids. This is the second find of this parasite originally described from hammerhead sharks off Australia, its first record from the western Atlantic Ocean, and its first record in a new host species and in hybrids.

First Myxozoan Infection (Cnidaria: Myxosporea) in a Marine Polychaete from North America and Erection of Actinospore Collective Group Saccimyxon.

In a survey of marine annelids for myxosporean infection in Charleston Harbor, South Carolina, we collected 3,214 polychaetes from 21 families and found infections in 6 spionid individuals. Based on gross morphology and COI sequencing, all infected spionids were identified as Streblospio benedicti. Infection prevalence was 0.8% (6/734) of that species of spionid, and 0.2% of all 3,214 polychaetes examined. Pansporocysts contained 8 actinospores and developed in the tegument of the annelid host. This is the first myxozoan infection recorded from this polychaete species, second in the family, and the first marine myxozoan found in the Americas. It is the first marine species found to develop in the tegument of its annelid host; a site of development observed only once before, in Ceratonova shasta infections of its freshwater sabellid polychaete host. Mature actinospores were morphologically simple, truncated ellipsoids, lacking processes or ornamentation, 9.0 ± 0.5 µm × 6.0 ± 0.4 µm. Their sack-like shape was similar to 9 of the 12 actinospores described previously from polychaetes; 10/12 had been and ascribed originally to the morphological collective group Tetractinomyxon despite 9 of these having few similarities to the original description of this group. We propose to name the simple, spherical to ellipsoidal spore morphotype Saccimyxon to encompass both our novel actinospore and the 9 other sack-like polychaete actinospore types in the literature. In the present study, 18S rDNA sequencing demonstrated that the myxozoans that infected the 6 spionids were genetically the same species (type sample 1,737 nucleotides, GenBank accession number MH791159) and was not >95% similar to any sequence in GenBank. Phylogenetic analysis showed that the myxozoan species we encountered is basal to the kudoids and thus likely to have a morphologically simple myxospore stage with fewer than 4 valves. However, without a genetic match, the presumptive vertebrate host remains unknown.

Description of Harpagorhynchus golvaneuzeti n. gen. n. sp. (Acanthocephala, Harpagorhynchinae n. sub-fam.) with a review of acanthocephalan parasites of soleid fishes in the Mediterranean Basin.

A species of acanthocephalan new to science from soleid fishes in the Mediterranean Sea and the Sea of Marmara is described. The new species is characterised by individuals having a club-shaped proboscis armed with 12-13 rows of 6-7 rooted hooks of a single type, a basal cerebral ganglion, and tegumental spines on the anterior two thirds of the body. Males have six cement glands and females show spines around the genital opening. To accommodate this species, a new genus, Harpagorhynchus n. gen., and a new subfamily in Echinorhynchidae, Harpagorhynchinae n. sub-fam., are erected. A critical review of the literature on echinorhynchid species infecting soleid fishes in the Mediterranean basin showed that Solearhynchus soleae (Porta, 1906) should be considered a junior synonym of S. rhytidotes (Monticelli, 1905) and that S. kostylewi (Meyer, 1932) is a valid species. An identification key of acanthocephalans of Mediterranean soleids is provided.

Redescription of three species of nematodes (Nematoda) parasitising fishes in the USA, with a key to the species of Dichelyne Jägerskiöld, 1902 parasitic in freshwater and brackish-water fishes of North America.

Three species of parasitic nematodes are redescribed based on light and scanning electron microscopical (SEM) examinations of newly collected specimens in fishes of South Carolina, USA: Dichelyne (Cucullanellus) bullocki Stromberg & Crites, 1972 from Fundulus heteroclitus (Linnaeus) (Fundulidae); Dichelyne (Dichelyne) diplocaecum Chandler, 1935 from Ictalurus furcatus (Valenciennes) (Ictaluridae); and Hysterothylacium pelagicum Deardorff & Overstreet, 1982 from Coryphaena hippurus Linnaeus (Coryphaenidae). For the first time, intraspecific variations in the number of intestinal caeca were observed in D. bullocki, as well as previously unknown males and gravid females of D. diplocaecum are described; this enabled to synonymise D. mexicanus Caspeta-Mandujano, Moravec & Salgado-Maldonado, 1999 with D. diplocaecum. Unlike most congeneric species, H. pelagicum has no double postanal papillae, as confirmed by SEM. A key to the species of Dichelyne Jägerskiöld, 1902 parasitic in freshwater and brackish-water fishes in North America is provided.

First evidence of polychaete intermediate hosts for Neospirorchis spp. marine turtle blood flukes (Trematoda: Spirorchiidae).

Life cycles of spirorchiids that infect the vascular system of turtles are poorly understood. Few life cycles of these blood flukes have been elucidated and all intermediate hosts reported are gastropods (Mollusca), regardless of whether the definitive host is a freshwater or a marine turtle. During a recent survey of blood fluke larvae in polychaetes on the coast of South Carolina, USA, spirorchiid-like cercariae were found to infect the polychaetes Amphitrite ornata (Terebellidae) and Enoplobranchus sanguineus (Polycirridae). Cercariae were large, furcate, with a ventral acetabulum, but no eyespots were observed. Partial sequences of D1-D2 domains of the large ribosomal subunit, the internal transcribed spacer 2, and the mitochondrial cytochrome oxidase 1 genes allowed the identification of sporocysts and cercariae as belonging to two unidentified Neospirorchis species reported from the green turtle, Chelonia mydas, in Florida: Neospirorchis sp. (Neogen 13) in A. ornata and Neospirorchis sp. (Neogen 14) in E. sanguineus. Phylogenetic analysis suggests that infection of annelids by blood flukes evolved separately in aporocotylids and spirorchiids. Our results support the contention that the Spirorchiidae is not a valid family and suggest that Neospirorchis is a monophyletic clade within the paraphyletic Spirorchiidae. Since specificity of spirorchiids for their intermediate hosts is broader than it was thus far assumed, surveys of annelids in turtle habitats are necessary to further our understanding of the life history of these pathogenic parasites.

Blood flukes Cardicola parvus and C. laruei (Trematoda: Aporocotylidae): life cycles and cryptic infection in spotted seatrout, Cynoscion nebulosus (Teleost: Sciaenidae).

Aporocotylidae comprises a diverse family of fish blood flukes, with adults found in blood or body cavity of marine, brackish, or freshwater fish. Aporocotylids are unique among the Digenea with many developing in polychaetes. The life cycle has been elucidated for only a few species that develop in polychaetes from marine/brackish environments and none for western Atlantic aporocotylids. The basis for this study was observations of blood fluke larvae in annelids from South Carolina, USA in 1982 prior to possible usage of molecular tools to specifically identify parasite larvae. Recent description of aporocotylid species and genotyping tools prompted revisiting original collection sites to examine polychaetes and fish as potential hosts. Polycirrid Enoplobranchus sanguineus and terebellids Amphitrite ornata, and Terebella lapidaria revealed infections with aporocotylid larvae. Adult blood flukes were also collected from fish commonly encountered in the same habitat: spotted seatrout (Cynoscion nebulosus), red drum (Sciaenops ocellatus), black drum (Pogonias cromis), and Atlantic croaker (Micropogonias undulatus). Sporocysts containing cercariae were found in individuals of each annelid species. Adult Cardicola parvus were found in spotted seatrout and Atlantic croaker, C. laruei in spotted seatrout, C. currani in red drum, and C. palmeri in black drum. Genotype analysis of ITS-2 and lsrDNA of all forms confirmed conspecific infections by C. parvus in E. sanguineus and A. ornata and C. laruei in T. lapidaria. This is the first description of complete life cycles of aporocotylids in the Western Atlantic and first evidence of cryptic infections of Cy. nebulosus with C. parvus.

Infection dynamics of Kudoa inornata (Cnidaria: Myxosporea) in spotted seatrout Cynoscion nebulosus (Teleostei: Sciaenidae).

Kudoa inornata is a myxosporean parasite that develops in the somatic muscle of spotted seatrout Cynoscion nebulosus, an economically and ecologically important fish in estuaries and harbors in southeastern North America. In South Carolina (SC), USA, over 90% of wild adult spotted seatrout are infected. To inform potential mitigation strategies, we conducted 3 experiments using naïve sentinel seatrout and infectious stages of K. inornata naturally present in raw water from Charleston Harbor, SC, to determine (1) if K. inornata infection follows a seasonal pattern, and (2) how long it takes for myxospores to develop in fish muscle. Infection by K. inornata was determined by visual detection of myxospores in fish muscle squashes, and any visually negative samples were then assayed for K. inornata ribosomal DNA using novel parasite-specific PCR primers. We observed that K. inornata infection in seatrout followed a seasonal pattern, with high prevalence when water temperature was highest (27-31°C; July-September) and infections that were either covert (at ~13-15°C) or not detected (<13°C) at the lowest water temperatures in January-February. Myxospore development occurred within 476 degree-days, i.e. 2 wk in a typical SC summer. Infection was dependent on fish density, which limited presumptive actinospore dose. Our findings suggest that the life cycle of the parasite may be disrupted by preventing spore-rich seatrout carcasses (e.g. at angler cleaning stations) being thrown back into harbors and estuaries throughout the year.

Pathogenic endoparasites of the spotted seatrout, Cynoscion nebulosus: patterns of infection in estuaries of South Carolina, USA.

Six types of pathogenic endoparasites in an economically important fish, spotted seatrout Cynoscion nebulosus, were studied in order to test whether prevalence of infection and assemblage richness varied with season, host sex, host size, or host age. Fish were collected from South Carolina estuaries, USA, over 12 months (n = 216; total lengths 15-663 mm). They were screened histologically for presence of Henneguya cynoscioni (Myxozoa) and Cardicola spp. (Digenea) in the heart, Kudoa inornata (Myxozoa) in the skeletal muscle, Sinuolinea dimorpha (Myxozoa) in the urinary system, Ichthyophonus sp. (Mesomycetozoea) in the kidney, and an unidentified microsporidian in the liver. Prevalence of infection was 29.8, 38.6, 47.2, 41.2, 13.6, and 2.8%, respectively. All factors had significant, but varying effects on the parasites. Parasite infections were more prevalent in winter than other seasons for Cardicola spp. and H. cynoscioni, more prevalent in winter and spring for Ichthyophonus sp., and more prevalent in male fish than female fish for K. inornata, S. dimorpha, and Ichthyophonus. Prevalence of infection by the three myxosporeans and Cardicola spp. increased with fish length, whereas prevalence of Ichthyophonus increased with length among young fish, but decreased with length among older fish. None of the factors affected the liver microsporidian, although statistical power was low due to its rareness. Assemblage richness varied between 0 and 5, was greater during winter and in male fish, and increased with fish length and fish age. Our results demonstrate that spotted seatrout are commonly co-infected by multiple pathogenic endoparasites, suggesting these parasites likely play an import role in controlling fish population numbers.

First Record of Paratenic Hosts of the Swimbladder Nematode Anguillicola crassus in North America.

Anguillicola crassus is a non-native parasite of the American eel, Anguilla rostrata. Since being introduced into North America, the nematode has spread rapidly across the range of A. rostrata, but paratenic hosts, which may facilitate parasite dispersion, have yet to be identified in the region. We investigated infection of larval A. crassus in 261 fish specimens belonging to 23 species and 12 orders collected from estuarine habitats in South Carolina (salinities 0-9 ppt) and Nova Scotia (10-18 ppt). A total of 35 fish belonging to 5 species and 3 orders were infected with the third-stage larvae (L3) of A. crassus, providing the first record of paratenic hosts for the parasite in North America. In South Carolina, high prevalence and abundance of the worm were found in spot (Leiostomus xanthurus), silver perch (Bairdiella chrysoura), and highfin goby (Gobionellus oceanicus), and a high prevalence but lower abundance was found in mummichog (Fundulus heteroclitus). In Nova Scotia, 2 nematodes were found in a single specimen of tomcod (Microgadus tomcod). All of the infected species are associated with a benthic lifestyle, and some of them are known to move between estuaries along the coastline. Lower infection rates in Nova Scotia may be associated with lower water temperatures and/or higher salinity of the sampling site. Most of the L3 were found encapsulated in mesenteric tissue around the intestine and stomach. No L4 or pre-adult worms were found. Mean body length of the L3 was smaller than L3 stages found in American eels from Cape Breton. This suggests that development of A. crassus is arrested at the L3 in the 5 fish species reported here, supporting their status as paratenic hosts.

The muscle dwelling myxozoan, Kudoa inornata, enhances swimming performance in the spotted seatrout, Cynoscion nebulosus.

Parasites usurp host resources and, as a consequence, enhance their transmission and increase their fitness while reducing the fitness of their host. Performance capacity is a key predictor of fitness. Thus, the effects of parasites on host fitness may often be mediated by alteration of host performance. We tested the effect of the skeletal muscle dwelling myxozoan, Kudoa inornata, on the swimming performance in spotted seatrout, Cynoscion nebulosus. We predicted greater infection would result in reduced swimming performance. Unexpectedly, increasing density of K. inornata myxospores in seatrout skeletal muscle was related to increased fish swimming performance. The experiment was repeated and confirmed these unexpected results. The mechanisms underlying enhanced performance of an infected host are not understood, but their occurrence emphasizes the role of parasites as selective pressures on host evolution.