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.

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.

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.

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.

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.

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.

Taxonomic notes on Polyacanthorhynchus kenyensis (Acanthocephala: Polyacanthorhynchidae) from Lake Naivasha, Kenya.

The original description of Polyacanthorhynchus kenyensis Schmidt and Canaris, 1967 from 2 species of fish in Lake Naivasha, Kenya was primarily based on measurements of 2 male cystacanths. That description is herein emended based on the availability of a larger number of immature worms of both sexes obtained from the freshwater teleosts Orechromis leucostictus, Tilapia zillii (new paratenic host records), and Micropterus salmoides. A secretory function for the apical organ is suggested.

Acanthocephala of marine fishes off Fiji Islands, with descriptions of Filisoma longcementglandatus n. sp., Neorhadinorhynchus macrospinosus n. sp. (Cavisomidae), and gravid females of Rhadinorhynchus johnstoni (Rhadinorhynchidae); and keys to species of the genera Filisoma and Neorhadinorhynchus.

A parasitological collection from marine fishes off the Fiji Islands yielded 5 species of acanthocephalans from 4 species of fish as follows: Filisoma longcementglandatus n. sp. from scat Scatophagus argus; Neorhadinorhynchus macrospinosus n. sp. from vermiculated rabbit fish Siganus vermicularis; Neorhadinorhynchus nudum (Harada, 1938) Yamaguti, 1939 (Cavisomidae) from mackerel tuna Euthynnus affinis; Rhadinorhynchus johnstoni Golvan, 1969 (Rhadinorhynchidae) from E. affinis; and Neoechinorhynchus agilis (Rudolphi, 1819) Van Cleave, 1916 (Neoechinorhynchidae) from striped mullet Mugil cephalus. All are new locality records and, except for N. agilis, new host records. Filisoma longcementglandatus is distinguished from all other species of the genus by its very long cement glands and size and armature of its proboscis. Neorhadinorhynchus macrospinosus has distinctly larger proboscis hooks than all other members of the genus. Keys to species of the genera Filisoma and Neorhadinorhynchus are included. Gravid females of the inadequately described R. johnstoni are described for the first time. Taxonomic notes on the other acanthocephalan species are also included.

Development and longevity of Nosopsyllus laeviceps kuzenkovi (Siphonaptera) from Inner Mongolia under laboratory conditions.

The development and longevity of Nosopsyllus laeviceps kuzenkovi (Yagubyants), a major plague vector among sylvan and campestral rodents in Inner Mongolia, China, was studied under controlled laboratory conditions of 11, 14, 17, 20, 23, 26, 29, and 35 C at 76% relative humidity (R.H.), and 32, 53, 76, 87, and 100% at 23 C. Development from egg to adult was shortest (21-23 days) under favorable conditions of 26 C at 76% R.H., but maximum survival to adulthood (84%) occurred at 23 C. Average longevity was 13.25 days (13.70 in females and 12.80 in males) at 17 C and 76% R.H.; it was longer at cooler and shorter at warmer temperatures. Findings provided experimental data that helped clarify seasonal fluctuations, summer mortality, generation cycles, and survival of compared flea species from Russia, Azerbaidzhan, and China.

Blood-feeding of the gerbil flea Nosopsyllus laeviceps kuzenkovi (Yagubyants), vector of plague in Inner Mongolia, China.

The gerbil flea Nosopsyllus laeviceps (Wagner) (Siphonaptera: Ceratophyllidae) is an important Palaearctic vector of enzootic plague among sylvan and campestral rodents. Laboratory blood-feeding experiments with the subspecies N.l.kuzenkovi (Yagubyants) from Inner Mongolia, China, using three rodent species including the natural host Meriones unguiculatus (Milne-Edwards), demonstrated a positive relationship between per cent feeding and time allowed for feeding, particularly for female fleas. Also the bloodmeal size was proportional to sucking time and was significantly greater in females, which are larger, than in males. However, on all four strains of host tested, there was no significant correlation between the feeding success rate of the fleas and the amount of blood they imbibed. Fleas fed better on active compared to confined M.unguiculatus. These findings agree with studies of other flea species.

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.

Description and pathology of Neoechinorhynchus idahoensis n. sp. (Acanthocephala: Neoechinorhynchidae) in Catostomus columbianus from Idaho.

Neoechinorhynchus idahoensis is described from Catostomus columbianus caught in the Salmon River, Stanley Basin, Idaho. The new species is closest to Neoechinorhynchus venustus Lynch, 1936, but is distinguished from it by its smaller and variably structured eggs, anterio-dorsal trunk hump, bent and posteriorly notched proboscis receptacle, and larger proboscis, proboscis receptacle, and hooks. It is distinguished also from 2 other species of Neoechinorhynchus with proboscis hooks in middle and anterior circles about equally large and from 7 other species having lemnisci greatly unequal in length. Histopathology of host tissue showed limited host response exemplified by epithelial damage and hemorrhaging at point of proboscis attachment with subsequent macrophage and other phagocytic cell migration. The proboscis extended through the host epithelium into the submucosa with limited hemorrhaging at the point of attachment. Unorganized collagenous fibers were present. The lumen of the host intestine was obstructed, and compressed villi were present. The trunk of the worm damaged intestinal epithelium near the crypts, causing localized inflammation. The caryophyllaeid cestode Isoglaridacris calentinei Mackiewicz, 1974, was present in concurrent infections of C. columbianus.

Description and host relationships of Polymorphus spindlatus n. sp. (Acanthocephala: Polymorphidae) from the heron Nycticorax nycticorax in Peru.

Polymorphus spindlatus n. sp. is described from the black-crowned night heron, Nycticorax nycticorax, in Lake Titicaca, Peru. It is distinguished from all 27 known species of the subgenus Polymorphus by its spindle-shaped proboscis and its trunk shape, the anterior 2/3 of which is ovoid, tapering into a tubular posterior end. It resembles Polymorphus brevis (=Arhythmorhynchus brevis), which is, however, longer and considerably more slender, and has smaller and more numerous proboscis hooks per row and smaller eggs. It is separated also from Polymorphus swartzi, Polymorphus striatus, Polymorphus contortus, and Polymorphus cincli by its proboscis armature (usually 18 longitudinal rows of 11-13 hooks each), among other characters. Histopathological sections of host tissue show well defined localized damage including hemorrhaging with subsequent phagocyte cell migration (granular tissue). The lumen of the host intestine is obstructed and villi show compression. The proboscis of P. spindlatus extends through the intestinal mucosa and submucosa, displacing the smooth muscle layers of the muscularis externa. Fibrosis also was observed.