The complete mitochondrial genome of 3 species of allocreadiids (Digenea, Allocreadiidae): characterization and phylogenetic position within the order Plagiorchiida.

Trematodes of the family Allocreadiidae are primarily found in the intestines of freshwater fishes around the world. The family includes 15 genera and c. 130 species. The last 2 decades have witnessed an increase in the genetic library of its species. Molecular data have been crucial for species delimitation and species description within Allocreadiidae and for understanding their evolutionary and biogeographical history and classification. Here, the mitogenomes of 3 species of allocreadiids were obtained using high throughput sequencing methods. Mitogenomes were compared with other members of the order Plagiorchiida to determine their molecular composition, gene rearrangement and phylogenetic interrelationships. The complete circular mitogenomes of Allocreadium lobatum, Creptotrematina aguirrepequenoi and Wallinia mexicana were 14 424, 13 769 and 13 924 bp long respectively, comprising 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and 2 non-coding regions. Gene arrangements were identical to other Xiphidiatan trematodes. Phylogenetic analyses using the mitogenomes revealed Allocreadiidae as a monophyletic group closely related to other members of the suborder Xiphidiata; A. lobatum was yielded as the sister taxon of C. aguirrepequenoi + W. mexicana. Our study increases the complete mitochondrial genome library of trematodes and strengthens our understanding of the phylogenetic relationships and classification of this parasite group.

Molecular evidence of three closely related species of Biacetabulum Hunter, 1927 (Cestoda: Caryophyllidea): a case of recent speciation in different fish hosts (Catostomidae)?

Monozoic tapeworms (Caryophyllidea) are dominant components of parasite communities of suckers (Catostomidae) in North America, with Biacetabulum Hunter, 1927 representing one of the more species-rich genera. Molecular (28S rDNA) and morphological (including scanning electron microscopy and histology) evaluation of newly collected tapeworms from different fish hosts revealed the existence of four similar (and three closely related) species of Biacetabulum. These four species differ from their congeners by having a long body (up to 48 mm long) with a very long, slender neck (its length represents ≥30% of total body length), a large, globular scolex with a prominent central acetabulum-like loculus on the dorsal and ventral sides, two pairs of shallow lateral loculi and a distinct, slightly convex apical disc, and a cirrus-sac that is situated between the anterior arms of the ovarian wings. Taken together, the morphological and molecular data and the host associations of these species provide evidence of their host specificity. Biacetabulum isaureae n. sp. occurs in notch clip redhorse, Moxostoma collapsum, in South Carolina (USA), B. longicollum n. sp. in silver redhorse, Moxostoma anisurum (type host), and golden redhorse, M. erythrurum, in Manitoba (Canada) and West Virginia (USA), B. overstreeti n. sp. in a spotted sucker, Minytrema melanops, in Mississippi, and B. hypentelii n. sp. in northern hogsucker, Hypentelium nigricans, in Tennessee (USA). The new species differ from each other in the number of postovarian vitelline follicles, the posterior extent of preovarian vitelline follicles and relative size of the cirrus sac.

The Proteocephalus species-aggregate (Cestoda) in cyprinoids, pike, eel, smelt and cavefish of the Nearctic region (North America): diversity, host associations and distribution.

Tapeworms (Cestoda) of the Proteocephalus-species aggregate from cyprinoids, pike, eel, smelt and cavefish in the Nearctic region (North America) are reviewed, based on a critical examination of newly collected and museum specimens. For some species neither new nor museum specimens were available and only literature data were used for these taxa. Two species occur in North American cyprinoids: (i) Nearctic Proteocephalus buplanensis Mayes, 1976 in Semotilus atromaculatus (Mitchill) in the upper Mississippi River and Hudson Bay drainage basins, and (ii) Holarctic P. torulosus (Batsch, 1786) (syns. P. ptychocheilus Faust, 1919; P. cobraeformis Haderlie, 1953; and Ophiotaenia critica Mpoame & Landers, 1981, new synonym), which occurs in Ptychocheilus spp. and other leuciscids in the western part of North America. Proteocephalus pinguis La Rue, 1911 is a specific parasite of pike (Esox spp.), which is briefly redescribed here to establish its diagnostic morphological characteristics because the original description was based on a mixture of at least two species. In addition to P. pinguis, pike serve as postyclic hosts of several species of the Proteocephalus-aggregate typical of other fish, such as bass, perch and whitefish, namely P. fluviatilis Bangham, 1925, P. pearsei La Rue, 1919 and P. longicollis (Zeder, 1800). Cavefish (Amblyopsidae) in North America harbour two endemic species, P. chologasteri Whittaker & Hill, 1968 from Chologaster agassizii Putman and P. poulsoni Whittaker & Zober, 1978 from Amblyopsis spelaea DeKay, which have never been found since their original description and their validity should be confirmed based on new collections and molecular data. Two other species of the Proteocephalus-aggregate, P. macrocephalus (Creplin, 1825), a parasite of eels (Anguilla spp.), and P. tetrastomus (Rudolphi, 1810) from smelt (Osmeridae), have circumboreal (Holarctic) distribution. Molecular data are available only for three of the seven species treated herein, but they do not form a monophyletic group.

Amended diagnosis, validity and relationships of the genus Acrolichanus Ward, 1917 (Digenea: Allocreadiidae) based on the 28S rRNA gene, and observations on its lineage diversity.

Adult specimens of the allocreadiid trematode Acrolichanus auriculatus (Wedl, 1858) were collected from Acipenser schrenckii Brandt from the River Amur and Amur Estuary, Acipenser ruthenus L. from the Rivers Irtysh and Oka (Asian and European parts of Russia, respectively), and Acipenser fulvescens Rafinesque from Lake Winnebago, Wisconsin, USA, and used for phylogenetic analyses based on 28S rDNA sequence data. The results supported the monophyly of Acrolichanus populations from both continents and a clear separation of A. auriculatus from other allocreadiid taxa, thus supporting the validity of the genus Acrolichanus Ward, 1917. This study formally reestablishes the genus Acrolichanus. Species of this genus differ from other allocreadiids by the following combination of morphological features: triangular mouth opening with rounded posterior corners, simple ventral, dorsolateral and dorsomedian muscular lobes on the oral sucker, stout cirrus-sac, pars prostatica with dorsal pocket, and massive cirrus with conspicuous triradiate lumen when invaginated. Specimens of the trematode from North America and the Rivers Irtysh and Oka were more closely related to each other than to the specimens from River Amur and Amur Estuary. This reflects the evolutionary divergence among sturgeons into Pacific and Atlantic clades. The phylogenetic analyses also confirmed the paraphyly of the genus Crepidostomum Braun, 1900.

Three new species of Plagioporus Stafford, 1904 from darters (Perciformes: Percidae), with a redescription of Plagioporus boleosomi (Pearse, 1924) Peters, 1957.

A form of Plagioporus Stafford, 1904 is described from the intestine of three North American species of darters (Perciformes: Percidae) from River West Twin, Wisconsin, USA, that we consider to be conspecific with Plagioporus boleosomi (Pearse, 1924) Peters, 1957 based on similarities in the sucker ratio, extent of the forebody, shape and position of the testes, vitellarium distribution and terminal genitalia. Three new species of Plagioporus are described from the intestine of darters as follows: Plagioporus fonti n. sp. from Percina nigrofasciata Agassiz in Florida, USA, Plagioporus limus n. sp. from Etheostoma squamosum Distler in Arkansas, USA and Plagioporus aliffi n. sp. from Etheostoma blennioides newmanni Miller in Arkansas, USA. Morphologically Plagioporus fonti n. sp., Plagioporus limus n. sp. and Plagioporus aliffi n. sp. are most similar to one another and to P. boleosomi, Plagioporus lepomis Dobrovolny, 1939 and 'P. etheostomae', a nomen nudum for a species described from Etheostoma blennioides Rafinesque in Kentucky, USA, all of which are collectively distinguished from congeners in having a combination of confluent vitellarium in the post-testicular space and absence of vitelline follicles with their entire length distributed in the forebody. Plagioporus fonti n. sp., P. limus n. sp. and P. aliffi n. sp. are respectively distinguished from one another and their closest congeners in having the anterior extent of the vitellarium in the anterior half of forebody to slightly anterior to the ventral sucker as opposed to one approximately at the level of the posterior margin of the ventral sucker, possession of an excretory vesicle reaching the anterior testis as opposed to one only reaching the posterior testis and having a longer than wide oral sucker and a wider than long ventral sucker. A Bayesian inference (BI) analysis of partial 28S rDNA sequences was conducted using the three new species and 24 sequences of opecoelids retrieved from GenBank, including ten species of Plagioporus. Plagioporus aliffi n. sp., Plagioporus fonti n. sp. and P. boleosomi comprised a moderately supported sister group to a clade containing all species of Plagioporus except Plagioporus limus n. sp. and Plagioporus shawi (Mcintosh, 1939) Margolis, 1970. Plagioporus limus and in turn P. shawi were resolved as sister to all other congeners with high and moderate support, respectively.

Morphological and molecular data for a new species of Pomphorhynchus Monticelli, 1905 (Acanthocephala: Pomphorhynchidae) in the Mexican redhorse Moxostoma austrinum Bean (Cypriniformes: Catostomidae) in central Mexico.

Pomphorhynchus purhepechus n. sp. is described from the intestine of the Mexican redhorse Moxostoma austrinum Bean (Catostomidae) in central Mexico. The new species can be distinguished from the other seven described species of Pomphorhynchus Monticelli, 1905 in the Americas by a subspherical proboscis and 14 longitudinal rows with 16-18 hooks each; the third and the fourth row of hooks are alternately longest. Sequences of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and the large subunit (LSU) rDNA (including the domains D2-D3) were used to corroborate the morphological distinction between the new species and Pomphorhynchus bulbocolli Linkins in Van Cleave, 1919, a species widely distributed in several freshwater fish species across Canada, USA, and Mexico. The genetic divergence estimated between the new species and the isolates of P. bulbocolli ranged between 13 and 14% for cox1, and between 0.6 and 0.8% for LSU. Maximum likelihood and Bayesian inference analyses of each dataset showed that the isolates of P. bulbocolli parasitising freshwater fishes from three families, the Catostomidae, Cyprinidae and Centrarchidae, represent a separate lineage, and that the acanthocephalans collected from two localities in central Mexico comprise an independent lineage. In addition, our analysis of the genetic variation of P. bulbocolli demonstrates that individuals of this acanthocephalan from different host species are conspecific. Finally, the distribution, host-association, and phylogenetic relationship of the new species, when placed in the context of the region's geological history, suggest that both host and parasite underwent speciation after their ancestors became isolated in Central Mexico.

A synoptic review of Promonobothrium Mackiewicz, 1968 (Cestoda: Caryophyllidea), parasites of suckers (Catostomidae) in North America, with description of two new species.

Monozoic cestodes of the recently amended genus Promonobothrium Mackiewicz, 1968 (Cestoda: Caryophyllidea), parasites of suckers (Cypriniformes: Catostomidae) in North America, are reviewed, with information on their host specificity, distribution and data on the scolex morphology of seven species studied for the first time using scanning electron microscopy (SEM). Evaluation of type and voucher specimens from museum collections and newly collected material of most species indicated the following valid nominal species: Promonobothrium minytremi Mackiewicz, 1968 (type species); P. ingens (Hunter, 1927); P. hunteri (Mackiewicz, 1963); P. ulmeri (Calentine et Mackiewicz, 1966); P. fossae (Williams, 1974) and P. mackiewiczi (Williams, 1974). Rogersus Williams, 1980 with its only species R. rogersi is transferred to Promonobothrium based on morphological and molecular data. Promonobothrium currani sp. n. and P. papiliovarium sp. n. are described from Ictiobus bubalus (Rafinesque) and Ictiobus niger (Rafinesque), and Erimyzon oblongus (Mitchill), respectively. The newly described species can be distinguished from the other congeners by the morphology of the scolex, the position of the anteriormost vitelline follicles and testes, the presence of postovarian vitelline follicles and the shape of the ovary. Molecular phylogenetic analyses of six species based on sequences of the small and large subunits of the nuclear ribosomal RNA genes (ssrDNA, lsrDNA) confirmed the monophyletic status of the genus and supported the validity of the species analysed. A key to identification of all species of Promonobothrium based on morphological characteristics is provided.

Trematode diversity in freshwater fishes of the Globe I: 'Old World'.

In this paper, we review, continent by continent, the trematode fauna of freshwater fishes of the 'Old World', a vast area consisting of the Palaearctic, Ethiopian, Oriental and Australasian zoogeographical regions. Knowledge of this fauna is highly uneven and clearly incomplete for almost all regions, sometimes dramatically so. Although the biggest problem remains the completion of the 'first pass' of alpha taxonomy, there are in addition great problems relating to biogeography and elucidation of life-cycles. For the latter, molecular data, i.e. matching DNA sequences of larval stages and corresponding adults, may represent a powerful tool that should be used in future studies. Another challenging problem represents the existence of cryptic species and, in particular, considerable decrease of experts in taxonomy and life-cycles of trematodes.

Trematode diversity in freshwater fishes of the Globe II: 'New World'.

We provide a summary overview of the diversity of trematode parasites in freshwater fishes of the 'New World', i.e. the Americas, with emphasis on adult forms. The trematode fauna of three regions, South America, Middle America, and USA and Canada (North America north of Mexico), are considered separately. In total, 462 trematode species have been reported as adults from the Americas. The proportion of host species examined for parasites varies widely across the Americas, from a high of 45% in the Mexican region of Middle America to less than 5% in South America. North and South America share no adult species, and one exclusively freshwater genus, Creptotrema Travassos, Artigas & Pereira, 1928 in the Allocreadiidae Looss, 1902 is the most widely distributed. Metacercariae of strigeiforms maturing in fish-eating birds (e.g. species of the Diplostomidae Poirier, 1886) are common and widely distributed. The review also highlights the paucity of known life-cycles. The foreseeable future of diversity studies belongs to integrative approaches and the application of molecular ecological methods. While opportunistic sampling will remain important in describing and cataloguing the trematode fauna, a better understanding of trematode diversity and biology will also depend on strategic sampling throughout the Americas.

A new species of Wallinia Pearse, 1920 (Digenea: Allocreadiidae) in Astyanax mexicanus (Characidae) from Mexico revealed by morphology and sequences of the 28S ribosomal RNA gene.

Wallinia mexicana sp. n. is described from the Mexican tetra, Astyanax mexicanus (De Filippi) (Characidae Weitzman), from two localities in northern Mexico. The new species can be distinguished from the two congeneric species, described from small-bodied characids in South and Central America, mainly by the posterior extent of the vitelline follicles (halfway between the posterior testis and the end of the caeca), by having a larger oesophagus, testes that are always oblique, and eye-spot remnants. The distinct status of the new species was confirmed by molecular data (28S rRNA gene sequences). Phylogenetic analysis suggests the new species is the sister species of W. chavarriae Choudhury, Hartvigsen et Brooks, 2002 described from characids in northwestern Costa Rica. Additionally, genetic divergence between these congeners reached 3.3%, a value higher than that observed for closely related species pairs of allocreadiids for that molecular marker. Based on these new findings, recently published records of this new species as Magnivitellinum simplex Kloss, 1966 and Creptotrematina aguirrepequenoi Jiménez-Guzmán, 1973 in Astyanax mexicanus from Durango and San Luis Potosi states, respectively, are corrected.

Gnathostoma spinigerum in live Asian swamp eels (Monopterus spp.) from food markets and wild populations, United States.

In Southeast Asia, swamp eels (Synbranchidae: Monopterus spp.) are a common source of human gnathostomiasis, a foodborne zoonosis caused by advanced third-stage larvae (AL3) of Gnathostoma spp. nematodes. Live Asian swamp eels are imported to US ethnic food markets, and wild populations exist in several states. To determine whether these eels are infected, we examined 47 eels from markets and 67 wild-caught specimens. Nematodes were identified by morphologic features and ribosomal intergenic transcribed spacer-2 gene sequencing. Thirteen (27.7%) M. cuchia eels from markets were infected with 36 live G. spinigerum AL3: 21 (58.3%) in liver; 7 (19.4%) in muscle; 5 (13.8%) in gastrointestinal tract, and 3 (8.3%) in kidneys. Three (4.5%) wild-caught M. albus eels were infected with 5 G. turgidum AL3 in muscle, and 1 G. lamothei AL3 was found in a kidney (both North American spp.). Imported live eels are a potential source of human gnathostomiasis in the United States.

ERECTION OF BOTHRIOCESTUS N. GEN. (CESTODA: BOTHRIOCEPHALIDEA) AND REDESCRIPTION OF BOTHRIOCESTUS CUSPIDATUS (COOPER, 1917) (SYN. BOTHRIOCEPHALUS CUSPIDATUS) FROM WALLEYE, SANDER VITREUS, (PERCIFORMES: PERCIDAE) IN NORTH AMERICA.

Based on previous molecular phylogenetic analyses, Bothriocestus n. gen. is erected to accommodate bothriocephalid tapeworms that have an elongate scolex, a well-developed apical disc, and a narrow neck region, parasitize freshwater fishes in the Holarctic, and were previously placed in the polyphyletic genus Bothriocephalus Rudolphi, 1808 (Cestoda: Bothriocephalidea). Bothriocestus claviceps (Goeze, 1782) n. comb., a parasite of eels (Anguilla spp.) in the Holarctic region, is designated as the type species. Another species of the new genus, Bothriocestus cuspidatus (Cooper, 1917) (syn. Bothriocephalus cuspidatusCooper, 1917) is redescribed from type and voucher specimens, and new material from the type host, the walleye, Sander vitreus (Mitchill, 1818) (Perciformes: Percidae), in Manitoba and Ontario (where the type locality is located) (Canada) and in New York state and Wisconsin. Bothriocestus cuspidatus of S. vitreus is characterized primarily by the possession of a narrow, long strobila (total length up to 18 cm) composed of distinctly craspedote, trapezoidal proglottids, with primary, secondary, and tertiary proglottids differing in size, and by an arrow-shaped (=cuspidatus) scolex that is distinctly broader than the first proglottids, widest near the base in lateral view and gradually becoming broader toward the anterior end in dorsoventral view. A "dwarf" form of B. cuspidatus (total length of 9-27 mm) from Johnny darter, Etheostoma nigrum Rafinesque, 1820, and tessellated darter, Etheostoma olmstedi Storer, 1842 (both Percidae: Etheostominae), is also characterized morphologically in the present paper.

REVISION OF SANGUINICOLA PLEHN, 1905 WITH REDESCRIPTION OF SANGUINICOLA VOLGENSIS (RASÍN, 1929) MCINTOSH, 1934, DESCRIPTION OF A NEW SPECIES, PROPOSAL OF A NEW GENUS, AND PHYLOGENETIC ANALYSIS.

Sanguinicola Plehn, 1905 comprises 26 species that collectively infect fishes from 8 orders (Cypriniformes, Characiformes, Siluriformes, Esociformes, Salmoniformes, Labriformes, Centrarchiformes, and Perciformes). Its revision is warranted because several species assigned to the genus could represent new genera, nucleotide sequences are wanting, many species have incomplete descriptions, and types for most species are missing or of poor quality. Herein, we emend Sanguinicola based on morphology and the first nucleotide-based phylogenetic analysis that includes multiple sequences from morphologically identified adult specimens. We describe Sanguinicola plehnae Warren and Bullard n. sp. from the heart of northern pike, Esox lucius Linnaeus, 1758 from Russia; provide supplemental observations of Sanguinicola volgensis (Rasín, 1929) McIntosh, 1934 from the heart of sabrefish (type species), Pelecus cultratus (Linnaeus, 1758) Berg, 1949 from Russia; describe Sanguinicola cf. volgensis from the heart of ide, Leuciscus idus (Linnaeus, 1758) Berg, 1949 from Russia; and describe Pseudosanguinicola occidentalis (Van Cleave and Mueller, 1932) Warren and Bullard n. gen., n. comb. from the heart of walleye, Sander vitreus (Mitchill, 1818) Bailey, Latta, and Smith, 2004 from eastern North America. Sanguinicola plehnae differs from its congeners by having lateral tegumental spines that total 118-122, are small (3% of body width), and protrude 2-3 µm from the tegument (lacking associated conical protrusion) as well as by having a large testis (>40% of body length). Sanguinicola volgensis differs from its congeners by having posteriorly directed lateral tegumental spines encased in a tegumental conical protrusion as well as by having an ovoid egg. Specimens of S. cf. volgensis differ from those of S. volgensis by having a body that is 5-6× longer than wide (vs. 2-3× in S. volgensis) and <90 lateral tegumental spines (vs. >95). Pseudosanguinicola Warren and Bullard n. gen. differs from Sanguinicola by having densely transverse rows of lateral tegumental spines (vs. a single column of large spines). The phylogenetic analysis utilizing the large subunit ribosomal DNA (28S) failed to reject monophyly of Sanguinicola.

A young parasite in an old fish host: A new genus for proteocephalid tapeworms (Cestoda) of bowfin (Amia calva) (Holostei: Amiiformes), and a revised list of its cestodes.

A new genus, Laruella n. gen., is proposed for the proteocephalid cestode L. perplexa (La Rue, 1911) n. comb. (syn. Proteocephalus perplexus La Rue, 1911), a parasite of a 'living fossil', the bowfin (Amia calva), in North America. The new genus is differentiated from other proteocephalid genera by having a massive four-lobed scolex without an apical organ and bearing suckers possessing tear-shaped sphincters on their inner rim, vitelline follicles forming L-shaped lateral fields, with the vitellarium turned inwards (medially) ventrally alongside the posterior margin of the ovary, a ring-like vaginal sphincter situated at a considerable distance from the genital atrium, and ellipsoid eggs resembling those of bothriocephalid and diphyllobothriid tapeworms, except for the absence of an operculum. Phylogenetic relationships of the new genus are not resolved, but it belongs to the so-called Neotropical clade of the Proteocephalidae, which is composed mainly of Neotropical tapeworms of siluriforms and other teleosts, but also Nearctic and Palaearctic species of Ophiotaenia La Rue, 1911 from snakes and amphibians. A morphologically similar species, Proteocephalus ambloplitis (Leidy, 1887) from bass (Micropterus spp.) in North America, is provisionally retained in Proteocephalus Weinland, 1858 because its relationships to L. perplexa are not yet clear. The former species differs from L. perplexa by the presence of a large apical organ, large, elongate vaginal sphincter situated near the genital atrium, vitelline follicles limited to lateral longitudinal fields, strongly coiled vas deferens within the cirrus sac, and a convoluted vaginal canal anterior to the ovarian isthmus. Laruella perplexa reportedly has a s broad spectrum of hosts but most are likely postcyclic or accidental hosts. A list of cestode parasites reported from bowfin is provided; it includes eight species and three taxa not identified to the species level. However, only three adult cestodes, L. perplexa and two species of Haplobothrium Cooper, 1914, are typical tapeworm parasites of bowfin, but previous molecular studies indicate possible existence of a putative new species in bowfin.

Host-Specific Parasites Reveal the History and Biogeographical Contacts of Their Hosts: The Monogenea of Nearctic Cyprinoid Fishes.

Host-specific parasites exhibit close co-evolutionary associations with their hosts. In the case of fragmented/disjunct host distribution, host-specific parasites may reflect the biogeographical history of regions and/or the role played by contacts of hosts. The present study was focused on Dactylogyrus (Monogenea) species almost exclusively parasitizing cyprinoid fishes. We investigated the phylogenetic relationships between Dactylogyrus parasites of Nearctic cyprinoids (Leuciscidae) and Dactylogyrus parasites of Palearctic cyprinoids and used Dactylogyrus phylogeny to explore the biogeography of fish hosts in Europe and North America. Phylogenetic analyses revealed that two Nearctic clades of Dactylogyrus spp. have different origins. Historical contacts between European and North American leuciscids were accompanied by the host switching of Dactylogyrus species. In the Nearctic region, Dactylogyrus parasites also colonized non-leuciscid fishes. Dactylogyrus spp. of three Nearctic leuciscid clades were included in the phylogenetic reconstruction; only Dactylogyrus spp. of the Plagopterinae had a common origin. Dactylogyrus species did not reflect the phylogenetic relationships among leuciscid clades, suggesting that past co-diversification was overshadowed by colonization events mediated by paleogeographic and climatological changes and extensive drainage reorganization. Host-specific monogeneans serve as a supplementary tool to reveal the historical biogeographical contacts between freshwater fish from the North America and Europe and also contemporary contacts of leuciscids in North America.

A NEW SPECIES OF BOTHRIOCEPHALUS (CESTODA: BOTHRIOCEPHALIDAE) FROM LEPOMIS SPP. (ACTINOPTERYGII: CENTRARCHIDAE) IN NORTH AMERICA.

Based on morphological and molecular data, a new species of tapeworm, Bothriocephalus kupermani n. sp., is described from pumpkinseed, Lepomis gibbosus (Linnaeus, 1758) (type host), and green sunfish, L. cyanellus Rafinesque, 1819 (Actinopterygii: Centrarchidae) in the United States. The new species differs from its North American congeners mainly in the shape of its scolex, which is almost rectangular in dorsoventral and lateral views (nearly parallel margins in the bothrial region), with shallow, wide bothria, and by an extensive field of vitelline follicles that are also variably present, albeit more sparsely, medially in the cortex of the ovarian and uterine areas. This tapeworm appears to be typically associated with Lepomis sunfishes. Examination of museum specimens of Bothriocephalus species from Lepomis spp. indicates that the tapeworm also parasitizes bluegill, Lepomis macrochirus, longear sunfish, Lepomis megalotis, and that previous records of Bothriocephalus cuspidatus and Bothriocephalus claviceps in these fishes are of the new species, B. kupermani.

LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR DETECTION OF ASIAN FISH TAPEWORM, SCHYZOCOTYLE ACHEILOGNATHI (YAMAGUTI, 1934) [SYN. BOTHRIOCEPHALUS ACHEILOGNATHI].

The Asian fish tapeworm (Schyzocotyle acheilognathi syn. Bothriocephalus acheilognathi) (AFT) is an invasive parasite that can infect many species of fish, although most hosts are primarily members of Cyprinidae. Pathogenicity has most often been reported in aquaculture settings in fry and fingerling stages of carp (Cyprinus spp.). More recently, it has been shown to cause growth retardation in the endangered bonytail chub (Gila elegans) and found to be widespread in populations of endangered humpback chub (Gila cypha) in the Colorado River, Grand Canyon, Arizona. AFT spreads most often through the transport of infected fish, particularly baitfish. Despite its harmful potential, there is no efficient or accurate ante mortem test to detect AFT in water or fish samples before transport. Herein, we report on the development of a sensitive and specific loop-mediated isothermal amplification (LAMP) assay to detect the parasite in under 30 min from laboratory prepared samples. Six LAMP primers were designed to amplify a variable region of the 18S ribosomal RNA gene in AFT with the detection and quantification of DNA on a real-time fluorometer. The limit of detection was 1 × 101 copies/µl of DNA extracted from as few as 2 AFT eggs. Future application of our assay would be a low-cost test to rapidly and accurately detect AFT DNA from environmental samples on-site so that preventive actions can be taken to halt the spread of the AFT through the movement of infected fish.

Ex Uno Plures? Morphotype and Lineage Diversity of Bothriocephalus (Cestoda: Bothriocephalidea) in North American Freshwater Fishes.

Morphological and molecular evaluation of tapeworms of the genus Bothriocephalus Rudolphi, 1808 (Cestoda: Bothriocephalidea), based on newly collected and uniformly fixed worms from freshwater fishes in Canada and the United States has revealed unexpected diversity. With a combination of selected morphological features and 4 molecular markers (18S rDNA V8 region, ITS1, ITS2, and COI gene sequences), the following morphotypes and lineages of the Bothriocephalus cuspidatus Cooper, 1917 complex were identified, several of which are specific to their respective fish definitive hosts and may represent separate species: B. cuspidatus sensu stricto from walleye, Sander vitreus (type host), which likely includes a miniature morphotype from Johnny darter, Etheostoma nigrum (both Percidae); Bothriocephalus morphotype from pumpkinseed, Lepomis gibbosus (Centrarchidae); and Bothriocephalus morphotype from rock bass, Ambloplites rupestris (Centrarchidae). The Bothriocephalus morphotype from goldeye, Hiodon alosoides (Hiodontidae), may also represent a separate lineage (possibly Bothriocephalus texomensisSelf, 1954) but requires additional studies. A morphotype from smallmouth bass, Micropterus dolomieu, based on a single specimen, is morphologically and genetically very similar to the morphotype from rock bass. Morphological study of the scolex and strobila of heat-killed and fixed specimens has revealed consistent differences, often subtle, that allowed us to differentiate between these morphotypes.

The Proteocephalus species-aggregate (Cestoda) in sticklebacks (Gasterosteidae) of the Nearctic Region, including description of a new species from brook stickleback, Culaea inconstans.

A survey of the species of the Proteocephalus-aggregate from sticklebacks (Actinopterygii: Gasterosteidae) is provided. The occurrence of three species in North America is confirmed: (i) Proteocephalus filicollis (Rudolphi, 1802), which has been reported from the three-spined stickleback, Gasterosteus aculeatus Linnaeus, in the northeastern part of North America (Newfoundland); (ii) Proteocephalus pugetensis Hoff et Hoff, 1929 occurs also in G. aculeatus, but in northwestern North America (British Columbia and Washington); and (iii) Proteocephalus culaeae sp. n., which is described from the brook stickleback, Culaea inconstans (Kirtland), in Manitoba (Canada). Another species, Proteocephalus ambiguus (Dujardin, 1845), a specific parasite of the nine-spined stickleback, Pungitius pungitius (Linnaeus), and type species of the genus, has also been found in North America (Alberta, Canada), but its vouchers are in poor condition and cannot be reliable assigned to this species. Both species reported from three-spined stickleback differ from each other by the shape of the scolex (rounded in P. filicollis versus continuously tapered towards the anterior extremity in P. pugetensis) and the apical sucker (widely oval to subspherical in frontal view in P. filicollis versus flattened in P. pugetensis). Proteocephalus culaeae sp. n. is characterised by a short body composed of a few, continuously widened proglottids, a short scolex narrower than the strobila and devoid of an apical sucker, a short, pyriform cirrus sac, no vaginal sphincter, and few testes. A key to species of the Proteocephalus-aggregate from sticklebacks is provided.

Tapeworms (Cestoda) of Ictalurid Catfishes (Siluriformes) in North America: Redescription of Type Species of Two Genera and Proposal of Essexiellinae n. Subfam.

Ictalurid catfishes (Siluriformes) in North America harbor proteocephalid tapeworms of the subfamily Corallobothriinae. Type species of 2 of 3 genera of these tapeworms from ictalurids are redescribed, based on museum and newly collected material. Essexiella fimbriata (Essex, 1928) is typified mainly by a wide, umbrella-shaped scolex with a metascolex formed by numerous folds of tissue, anteriorly directed suckers without sphincters, vitellarium bent inwards posteriorly, "flower-shaped" uterus (with anterior, lateral, and posterior diverticula), and a conspicuously pre-equatorial genital atrium. Verified host records of this cestode are only from 3 species of Ictalurus Rafinesque, 1820. Megathylacoides giganteum (Essex, 1928), which seems to be specific to the channel catfish (Ictalurus punctatus), possesses a globular scolex, with a weakly developed metascolex formed by tissue folds posterior to the suckers, anterolaterally directed suckers with large semilunar sphincters, proglottids that are widest at the level of the genital atrium at the anterior third of the proglottid, and uterine diverticula that do not reach the vitelline follicles laterally. A new subfamily, Essexiellinae Scholz and Barcák, is proposed to accommodate species of EssexiellaScholz, de Chambrier, Mariaux and Kuchta, 2011 (type genus), MegathylacoidesJones, Kerley and Sneed, 1956, and CorallotaeniaFreze, 1965 from ictalurid catfishes in the Nearctic Region. These tapeworms possess a metascolex, medullary genital organs, uterus lined with numerous chromophilic cells, pre-equatorial genital atrium, and uterine development of type 2. The new subfamily was monophyletic in all molecular phylogenetic analyses, being most closely related to 3 Neotropical proteocephalids from the redtail catfish, Phractocephalus hemioliopterus (Bloch and Schneider, 1801), but distant from all remaining proteocephalid tapeworms from freshwater fishes in North America.