The elevation of a unique population of Corynosoma strumosum (Acanthocephala: Polymorphidae) from the Caspian seal, Pusa caspica, in the Caspian Sea to Corynosoma neostrumosum n. sp.

An isolated population of 700 specimens initially described as Corynosoma strumosum (Rudolphi, 1802) Lühe, 1904 and currently reassigned to Corynosoma neostrumosum n. sp. was collected from one young male Caspian seal, Pusa caspica (Gmelin) in the southern land-locked Caspian Sea in April 2009. Collected worms were morphologically unique compared with those reported by other observers in open waters, especially in shape and distribution of proboscis hooks and trunk spines, dorso-ventral differences in proboscis hooks and their organization, the baldness of anterior proboscis, consistently smaller size of trunk and testes, larger eggs, the rough egg topography, epidermal micropores, and variations in the female gonopore. Molecular data from the internal transcribed spacer region of rDNA and the mitochondrial cox1 gene was also provided to supplement the morphological study of the new species.

Revision of Corynosoma australe Johnston, 1937 (Acanthocephala: Polymorphidae) from a North American population using novel SEM images, Energy Dispersive X-ray Analysis, and molecular analysis.

We describe a population of the acanthocephalan Corynosoma australe Johnston, 1937 (Polymorphidae) from a California sea lion Zalophus californianus (Lesson, 1828) in California using novel scanning electron microscopy (SEM) images, Energy Dispersive x-ray analysis (EDXA), and molecular analysis for the first time. The taxonomic history of C. australe is replete with accounts using only line drawings some of which proved erroneous. The distribution of ventral spines on the female trunk has been the primary distinction between C. australe and Corynosoma obtuscens Lincicome, 1943, its junior synonym; being continuous in the latter but discontinuous posteriorly in the former species. The distribution of ventral spines is invariably discontinuous in males. Our redescription and SEM images help to resolve this issue further validating the synonymy. Morphological variability has been documented between our California population and others from various host species in California, South Australia, South Shetlands, and the Argentinian coast. Our SEM images document features not previously detectable in line drawings, erroneously reported or missed in previous accounts. The EDXA spectra show high levels of calcium and phosphorous and low levels of sulfur characteristic of C. australe. EDXA for other species of Corynosoma Lühe, 1904 provide support for the diagnostic distinction of C. australe. EDXA spectra were shown to be species specific and have diagnostic value in the taxonomy of the Acanthocephala. Our molecular analysis used amplification of 18S of ribosomal DNA and cytochrome c oxidase 1 (Cox1) gene. Phylogenetic analyses for Cox1 gene revealed a close relationship between Corynosoma hannae Zdzitowiecki, 1984 and C. australe. The phylogenetic trees confirmed that the isolates belonged to C. australe. The haplotype network inferred by Cox1 with C. australe sequences revealed that haplotypes clearly separated from each other and formed clusters related to samples from the Northern Hemisphere (the USA and Mexico), and the second from the Southern Hemisphere (Argentina, Brazil and Peru).

New perspectives of Microsentis wardae Martin & Multani, 1966 (Acanthocephala: Neoechinorhynchidae) from Gillichthys mirabilis Cooper in California, with scanning electron microscopy images and energy dispersive X-ray analysis.

Fully developed, sexually mature small male and female acanthocephalans, Microsentis wardae Martin & Multani, 1966 (Neoechinorhynchidae) reaching only 2.25 mm and 2.42 mm, respectively, were collected from the rectum of longjaw mudsuckers, Gillichthys mirabilis Cooper, in the salty marches of Anaheim Bay and San Diego Bay, California. Our specimens were half the size of those reported in the original description from the same host in Scammon's Lagoon over 700 km to the south. The ratio of proboscis and receptacle size to trunk size was markedly higher in our specimens compared to the larger specimens measured in the original description. The anatomy of all structures in our specimens from Anaheim Bay was comparable to that of the larger Scammon's Lagoon specimens that have apparently realized more growth in the Scammon's Lagoon. We have observed more structures that are not reported in the original description, especially evident from our scanning electron microscopy images, which are not possible to observe in the original line drawings. In our specimens, the micropores were unusually widely spaced and the energy dispersive X-ray analysis showed longitudinal hook sections with high levels of sulphur and phosphorus and moderate levels of calcium, but the whole hooks showed highest levels of sodium and magnesium - the biochemical hook signature of this species. Sequences of the small subunit (18S) of the nuclear ribosomal DNA were generated and compared with acanthocephalan sequences available from GenBank. As M. wardae comprises a monotypic genus, therefore, phylogenetic analyses inferred from the 18S gene showed its relationship with other species of closely related genera of Eoacanthocephala. This is the first report of molecular data of M. wardae.

Redescription and molecular analysis of Pallisentis (Pallisentis) nandai Sarkar, 1953 (Acanthocephala: Quadrigyridae) in India.

Pallisentis (Pallisentis) nandai Sarkar, 1953 is a freshwater fish parasite restricted to the Indian subcontinent in the Ganga River and its tributaries. It was described from the leaffish, Nandus nandus (Hamilton) from the Ganga River delta at Calcutta. We recovered variant specimens from the same host species from the Ganga near its headwaters at Bijnor about 1500 km away. Our specimens were clearly identifiable as P. nandai but varied considerably from those in the original description, especially in the size of proboscis hooks, receptacle and lemnisci. The original description was incomplete (missing line drawings of female trunk and reproductive system, male trunk, complete proboscis, hooks and hook roots) and inaccurate (proboscis, hooks, receptacle wall), and some measurements were lumped together for both sexes. We provide a complete description and include new morphological information including the first description of para-receptacle structure in the genus Pallisentis Van Cleave, 1928, scanning electron microscopy and microscope images, molecular analysis, and energy dispersive X-ray analysis (EDXA) of hooks and spines of our specimens for the first time. Additional details of proboscis hook roots, trunk spines, micropores and micropore distribution are described. The unique metal composition of hooks (EDXA) demonstrated a considerably high but variable level of sulphur and negligible level of calcium in collar and trunk spines and hook tips, but a higher level of sulphur and calcium at the hook basal arch than at the hook tip and edge. A comparison with the EDXA pattern of another species of Pallisentis, P. Indica Mital & Lal, 1976, were considerably different. The phylogenetic position of P. nandai within Eoacanthocephala was generated to assess the molecular characterization based on 18S and ITS1-5.8S-ITS2 ribosomal DNA sequences. Maximum likelihood and Bayesian inference analyses placed P. nandai in a clade with other Pallisentis species under the family Quadrigyridae. This is the first report based on molecular evidence for P. nandai.

New perspectives on Aspersentis Megarhynchus (Acanthocephala: Heteracanthocephalidae) from Notothenia Coriiceps Richardson (Nototheniidae) in the West Antarctic, with emended generic diagnosis.

A number of variable descriptive accounts of Aspersentis megarhynchus (von Linstow, 1892) Golvan, 1960 have been reported from specimens collected from many species of fish in various locations off Antarctic islands. We have described a new population from Notothenia coriiceps Richardson (Nototheniidae) off Galindez Island, West Antarctica, and features not previously reported, resolved the taxonomic controversies and nomenclature, and emended and updated the generic diagnosis taking into account the newly observed structures. These are depicted in microscopic images and include the outer spiral wall of the proboscis receptacle, the thicker dorsal wall of the receptacle compared to the ventral wall, parts of the female reproductive system, the separate cement gland ducts, the dorsal position of the male gonopore and more detail of proboscis hooks and trunk spines. It is surprising that the newly observed features were missed from the many descriptions of A. megarhynchus created since the original description. The variability in A. megarhynchus is noted with a comparison of the morphometrics of our specimens vs. those in six other descriptions. We also analysed the metal composition of hooks and spines using energy-dispersive X-ray analysis and concluded a molecular characterization of the species based on 18S DNA gene, with related phylogenetic analyses.

Description and molecular analysis of an Italian population of Centrorhynchus globo caudatus (Zeder, 1800) Lühe, 1911 (Acanthocephala: Centrorhynchidae) from Falco tinnunculus (Falconidae) and Buteo buteo (Accipitridae).

Centrorhynchus globocaudatus (Zeder, 1800) Lühe, 1911 (Centrorhynchidae) was reported in birds of prey. Our population from Falco tinnunculus Linnaeus (Falconidae) and Buteo buteo Linnaeus (Accipitridae) in northern Italy was morphologically distinct from others described elsewhere. The worms are elongate and cylindrical. Proboscis long, apically truncated and bare, with wider base and variably faint constriction at point of attachment of receptacle. Large anterior hooks well rooted; posterior spiniform hooks with reduced roots; transitional hooks with scutiform roots in-between. Four tubular cement glands extend into prominent ducts overlapping a large Saefftigen's pouch. Bursa large, with sensory plates. Vagina with laterally slit orifice in sub-ventral pit of globular terminal extension. Thick-shelled eggs ovoid without polar prolongation of fertilization membrane. In our specimens, proboscis hooks, receptacle, male reproductive system, and lemnisci especially in males varied in size from those from Ukraine, India, Egypt, Kyrgystan, Russia, Georgia, Armenia and Asian Soviet Republics. Our description of the Italian specimens includes new morphological information supported by scanning electron microscopy and microscope images, molecular analysis and energy dispersive X-ray analysis (EDXA) of hooks. Additional new details of proboscis hook roots, micropores and micropore distribution are described. Metal composition of hooks (EDXA) demonstrated high levels of calcium and phosphorous, and high levels of sulphur in core and cortical layers of eggs. The molecular profile based on sequences of 18S and cytochrome c oxidase 1 genes is also provided, as well as phylogenetic reconstructions including all available sequences of the family Centrorhynchidae, although further sequences are needed in order to clarify their phylogenetic relationships.

New perspectives on Nephridiacanthus major (Acanthocephala: Oligacanthorhynchidae) collected from hedgehogs in Iran.

We describe morphological features not previously reported for this old acanthocephalan Nephridiacanthus major (Bremser, 1811 in Westrumb, 1821) Golvan, 1962 first described over 200 years ago. Our specimens were collected from long-eared hedgehog Hemiechinus auritus (Gmelin, 1770) (Erinaceidae) in Iran. We compare the morphometrics of our material with others previously reported from the same host in Iran, Russia, central Asia and Europe. Our specimens had markedly smaller proboscides, proboscis hooks and lemnisci than those reported from Russia and central Asia, but comparable measurements of other structures with specimens previously described from other collections. We document our new observations with scanning electron microscopy features not previously demonstrable by other observers and provide a chemical analysis of proboscis hooks using energy-dispersive X-ray analysis for the first time. The molecular profile of this acanthocephalan, based on 18S rDNA and cox1 genes, was generated for the first time. The phylogenetic analysis showed that N. major is placed in a clade of the family Oligacanthorhynchidae, well separated from the families Moniliformidae and Gigantorhynchidae.

The Molecular Profile of Paratrajectura Longcementglandatus Amin, Heckmann Et Ali, 2018 (Acanthocephala: Transvenidae) from Percid Fishes in the Marine Waters of Iran and Iraq.

Paratrajectura longcementglandatus Amin, Heckmann et Ali, 2018 (Transvenidae) was recently described from two species of percid fishes collected from the marine territorial waters of Iraq and Iran in the Persian Gulf. The genus Paratrajectura Amin, Heckmann et Ali, 2018 is a close relative to transvenid genera Trajectura Pichelin et Crib, 2001 and Transvena Pichelin et Crib, 2001. Morphologically, Paratrajectura is characterised by having apical proboscis cone, long, tubular cement glands, short lemnisci, prominent roots on all proboscis hooks, subterminal female gonopore, and males with long pre-equatorial testes. Molecular studies of P. longcementglandatus using 18S rDNA and cox1 genes compared with available data of members of other families of Echinorhynchida showed that P. longcementglandatus is grouped with species of the genus Transvena forming a clade within the family Transvenidae.

Expanded description of Neoechinorhynchus (Hebesoma) manubrianus (Acanthocephala: Neoechinorhynchidae) from marine fish in Halong Bay, Vietnam.

Neoechinorhynchus manubrianus Amin, Ha & Ha, 2011 (Acanthocephala: Neoechinorhynchidae) (formerly Neoechinorhynchus manubriensis Amin, Ha & Ha, 2011), was recently described based on optical microscopy of four males and two females (none was gravid) from caroun croaker, Johnius carouna (Cuvier), flower croaker, Nibea albiflora (Richardson), and silver croaker, Pennabia argentata (Houttuyen) (Sciaenidae) in Halong Bay, Vietnam. Subsequently, many more specimens became available from N. albiflora that were studied using SEM. SEM studies showed many additional features that were not possible to discern with optical microscopy. These included the prominent angulation of the anterior trunk, the presence of (1) anterio-dorsal and (2) undulating mid-lateral fin-like protrusions of the body wall, uniquely shaped eggs as well as details of trunk micropores, proboscis, bursa, and female gonopore. Microscopical examination of eggs from the new collection demonstrated the presence of polar prolongation of fertilization membrane which places N. manubriensis in the subgenus Hebesoma. The features of trunk angulation, trunk fins, and egg morphology further distinguish N. manubriensis from all other species of Neoechinorhynchus Stiles and Hassall, 1905 from Vietnam or from any where else in the world.

First report of Neoechinorhynchus (Acanthocephala: Neoechinorhynchidae) from marine fish of the eastern seaboard of Vietnam, with the description of six new species.

The occurrence of acanthocephalans of the genus Neoechinorhynchus Stiles and Hassall, 1905 in Vietnamese waters is reported for the first time. Six new species are described from seven species of marine fish of the families Belonidae, Clupeidae, Megalopidae, Mugilidae, and Sciaenidae, collected in Halong Bay of the eastern seaboard of Vietnam in 2008 and 2009. These are Neoechinorhynchus (Neoechinorhynchus) plaquensis n. sp. characterized by dermal plaques covering the entire trunk; Neoechinorhynchus manubriensis n. sp. with very long anterior proboscis hooks having roots with prominent anterior manubria and very small and equal middle and posterior hooks, two pseudo-retractors in the receptacle, simple vagina, and terminal gonopore; Neoechinorhynchus pennahia n. sp. with equal anterior and middle proboscis and somewhat smaller posterior hooks, and terminal female gonopore; Neoechinorhynchus ampullata with many giant nuclei in the body wall and lemnisci and a parareceptacle structure complex which includes pumping ampullas reported for the first time; Neoechinorhynchus (Neoechinorhynchus) longinucleatus n. sp. with very long giant nuclei in the Lemnisci, anteriorly twisted vagina, and subterminal female gonopore. Neoechinorhynchus (Neoechinorhynchus) ascus n. sp. is the second species of Neoechinorhynchus found with the parareceptacle structure/ampulla complex. Neoechinorhynchus (Neoechinorhynchus) johnii Yamaguti, 1929 of Bilqees, 1972 is not N. johnii because of proboscis armature and other discrepancies with the Yamaguti material. Notes on host distribution and feeding habits are also included.

The molecular profile of Acanthogyrus (Acanthosentis) kashmirensis from the Indian subcontinent.

PURPOSE: The purpose of the present study was to provide molecular support for the validity of the morphological description of Acanthosentis kashmirensis Amin, Heckmann, Zargar, 2017 which was originally poorly described as Neoechinorhynchus kashmirensis Fotedar and Dar, 1977 from the Indian subcontinent, and to characterise its molecular identity and phylogenetic relationships. METHODS: Total DNA was extracted, and the partial region of the small subunit (SSU) 18S rDNA and ITS-rDNA genes were amplified and sequenced. Genetic diversity was calculated and phylogenetic analysis of the nucleotide sequence data was performed. RESULTS: In this study, the molecular profile of this acanthocephalan was generated for the first time. Based on the partial 18S rDNA, interspecific variation between A. kashmirensis with different species of Acanthosentis and were 3.6-10.3% and 22.4-38.2% based on 18S rDNA and ITS-rDNA genes, respectively. We described the phylogenetic relationships of A. kashmirensis compared with other species of the genus and also with members of the family Quadrigyridae. CONCLUSIONS: The ITS-rDNA sequences of members of the family Quadrigyridae are more variable than 18S rDNA that can be useful for achieving a proper assessment of biodiversity. Sequence data generation from additional species of Acanthosentis will be needed to reconstruct the phylogenetic relationships of this group of acanthocephalans.

Evaluation of a new system for the fixation, concentration, and staining of intestinal parasites in fecal specimens, with critical observations on the trichrome stain.

Proto-fix (Alpha-Tec Systems, Inc., Vancouver, WA) is a new single vial, environmentally safe, parasitology (pathogenic and nonpathogenic protozoans and helminths) fixative and transport solution. It is used in conjunction with a new concentration/sedimentation reagent, CONSED, (Alpha-Tec Systems, Inc. Vancouver, WA) as a replacement to the formalin-ethyl acetate (FEA) concentration procedure using Lugol's iodine. The newly adopted procedure was tested against the FEA concentration samples using split proficiency testing samples supplied by the American Association of Bioanalysts (AAB). Routinely, patient samples collected, fixed, and transported in Proto-Fix were processed and tested at Diagnostic Labs, Inc. (DLI), Phoenix, AZ. Detected parasites were documented using a video camera-printer system attached to the optical equipment. The quality of the fixative and stain were found to be superior to that of the FEA-Lugol's method and the yield of detected parasites was considerably higher. Eighty-five percent of 39 unknown parasite species tested were correctly detected using the Proto-fix-CONSED system compared to 46% using the FEA-Lugol's method. Of all the other methods and stains used at DLI, the trichrome stain (a popular modification of Gomori's trichrome stain for tissue sections) was found to be least reliable.

Acanthocephala from lake fishes in Wisconsin: ecology and host relationships of Pomphorhynchus bulbocolli (Pomphorhynchidae).

Pomphorhynchus bulbocolli Linkins in Van Cleave, 1919, was considerably more common in fishes of the river-connected Tichigan Lake than of the landlocked Silver Lake, southeastern Wisconsin. It is reported from 17 species of principal, accessory, and occasional definitive hosts (new record in Moxostoma carinatum) and from 13 species of paratenic hosts (new records in Amia calva, Ictalurus punctatus, Lepomis cyanellus, and Pomoxis nigromaculatus). Infection patterns were influenced by fish species, feeding behavior, temperature, availability of intermediate host, type of water body, fish movement, and changes in fish host community. Host roles are not fixed but are often interchangeable. A seasonal cycle in prevalence, intensity, and maturation was evident, with greatest abundance and maturation during summer and recruitment during summer and autumn. Recruitment of new infections, development, and release of eggs, however, occurred all year. Sex ratio changed from near equal in new infections to one more highly in favor of females in older adults. Female fish were considerably more frequently and heavily infected than males. No relationship with fish age (size) was evident. Worms were mostly attached in posterior intestinal locations but initial establishment sites correlated with temperature. Translocation of P. bulbocolli due to competitive exclusion in concurrent infections was not observed. The significance of extraintestinal larval forms in the cycle of transmission was noted.

Redescription of Hebesoma violentum Van Cleave, 1928 (Acanthocephala: Neoechinorhynchidae).

Hebesoma violentum Van Cleave, 1928, is redescribed from 155 paratypes from Van Cleave's original 1921 and 1923 collections in China. The present distribution of H. violentum in China and the Soviet Union is reported and the uncertain status of Hebesoma derosum Gupta and Jain, 1973, examined.

Key to the families and subfamilies of Acanthocephala, with the erection of a new class (Polyacanthocephala) and a new order (Polyacanthorhynchida).

A key to the classes, orders, families, and subfamilies of Acanthocephala based on the Meyer-Van Cleave system as presented by Amin (1985) is provided. A new class (Polyacanthocephala) and a new order (Polyacanthorhynchida) are erected to accommodate members of the monogeneric family Polyacanthorhynchidae Golvan, 1956.

Fessisentis tichiganensis sp. nov. (Acanthocephala: Fessisentidae) from Wisconsin fishes, with a key to species.

Fessisentis tichiganensis sp. nov. is described on the basis of 16 male and 24 female worms recovered from 11 species of fish of seven families in Tichigan Lake, southeastern Wisconsin, between 1977 and 1979. Specimens were most frequently encountered during the spring; the only two gravid females were recovered from Umbra limi, in early June. The same fish also serves as a transport host. A key separating F. tichiganensis from the four other known species in North America is presented. The new species most closely resembles F. friedi Nickol 1972 (= F. vancleavei haley and Bullock 1953), but can be separated from it by its longer proboscis which is armed with markedly more hooks per row, among other characteristics. Additional, new morphological observations are also reported.

Acanthocephalus parksidei sp. n. (Acanthocephala: Echinorhynchidae) from Wisconsin fishes.

Acanthocephalus parksidei sp. n. is described from over 4,000 specimens recovered from 11 fish hosts in the Pike River, southeastern Wisconsin. A parksidei is a highly variable acanthocephalan which appears to present an intermediate form between the only 2 other members of the same genus, A. dirus and A. jacksoni, described from North American freshwater fishes. It is structurally similar to the larger A. dirus but is separated from it primarily by having fewer proboscis hook rows and hooks per row. It is separated from A. jacksoni on the basis of its slender body form, relatively larger size of male reproductive system in proportion to body size, as well as other features. It differs from the larger European A. lucii by having smaller proboscis hooks, different hook root shape, smaller proboscis receptacle, and smaller embryos.

Variability in Acanthocephalus parksidei Amin, 1974 (Acanthocephala: Echinorhynchidae).

Acanthocephalus parksidei Amin, 1974, was recently described from 11 species of Wisconsin fishes. Considerable variation was observed in this acanthocephalan's body size, number and arrangement of proboscis hook, size of hooks, proboscis, proboscis receptacle, lemnisci, embryos, testes, and cement glands, and number of the latter two. Most of these variations were related to worm sex and age as well as host species. Structural variations in the shape and lobation of lemnisci, abnormalities in hooks and body wall, and changes associated with monorchidism in males were noted. The significance of the erection of this new taxon in relation to the taxonomic criteria applied to acanthocephalan species is discussed.

Host and seasonal associations of Acanthocephalus parksidei Amin, 1974 (Acanthocephala: Echinorhynchidae) in Wisconsin fishes.

Ecological associations of Acanthocephalus parksidei Amin, 1974, were studied in 11 southeastern Wisconsin fishes during 1972-73. Parasitic load was found to be related to host species, size (age), sex, concurrent infections, collection site, and season. Distribution in host intestine was related to the extent or intensity of infection, posterior migration, and concurrent infections. Parasitic cycle commenced in later summer-early autumn, peaked (in frequency and maturation) during the spring, and terminated in the late summer.