A new species of Hedruris (Nematoda: Hedruridae) from freshwater turtles, its life cycle and biogeographic distribution of the genus.

We describe Hedruris dratini n. sp. (Nematoda, Hedruridae) from Hydromedusa tectifera and Phrynops hilarii in Argentina based on morphological and molecular characters. Also, we provide information about its life cycle. The new species differs from other species of the genus by possessing the excretory pore, nerve ring and deirids at equal distance from the anterior end. Additionally, H. dratini n. sp. has mammilated eggs and males possess nine pairs of caudal papillae. The subadults and adults of H. dratini n. sp and H. orestiae were characterized by sequencing the small subunit ribosomal DNA (18S). We present for the first time a life cycle of a species of Hedruris that includes an amphipod as intermediate host and a reptile as definitive host. Furthermore, we analysed the host and geographic distribution of all Hedruris species. Although the genus has a cosmopolitan distribution and parasitizes a great host diversity, the majority of species have a Gondwanian distribution, with amphibians being the preferred hosts.

Larval spirurids in a supralittoral amphipod in the north-east of Russia and the identification of the intermediate host of Antechiniella septentrionalis (Spirurida: Acuariidae), parasitic in a tundra vole.

The supralittoral amphipod Traskorchestia ditmari (Derzhavin, 1923) was identified as the intermediate host for Antechiniella septentrionalis Ivanova, Dokuchaev & Spiridonov, 2019, a parasite of the tundra vole Microtus oeconomus and Skrjabinocerca sp. (both Spirurida: Acuariidae) in Magadan Oblast in north-eastern Russia. Joint infection by both larval spirurids was not observed. The infective stage of A. septentrionalis was the encysted larvae, while larvae of Skrjabinocerca sp. were free in the amphipod's coelom. The identity of A. septentrionalis was confirmed using cox1 mtDNA gene analysis, performed on adult stages from a tundra vole and on larvae from amphipods. Possible transmission routes of A. septentrionalis are discussed.

A new species of Procyrnea (Nematoda: Habronematidae) parasitic in Rhea pennata (Aves: Rheidae) from Patagonia, Argentina, with a key to species of the genus.

Procyrnea choique n. sp. is described from the lesser rhea, Rhea pennata d´Orbigny (Aves: Rheidae), from the Protected Natural Area Península Valdés, Chubut province, Argentina. The new species resembles P. ficheuri, P. murrayi, P. excisiformis, P. dollfusi, P. haliasturi, P. anterovulvata, P. graculae, P. brevicaudata, P. uncinipenis, P. javaensis, P. ameerae, P. ornata, P. aegotheles, P. spiralis, P. ruschii and P. aptera, mainly in the absence of lateral alae; but differs from its congeners by having a left spicule without barbs, the position of the vulva which is post-equatorial, the absence of lateral ridges, absent median precloacal papilla, asymmetrical caudal alae, males more than 6 mm long, left spicule 1 mm long and spicule ratio 1:3. Our results extend the taxonomy of Procyrnea Chabaud, 1958 and comprise the first report of a habronematid from R. pennata. A key to species of Procyrnea is presented.

Use of molecular tools in identification of philometrid larvae in fishes: technical limitations parallel our poor assessment of their biodiversity.

The usefulness of the polymerase chain reaction with restriction fragment length polymorphism (PCR-RFLP) and partial sequencing of the cytochrome oxidase I (COI) gene was tested regarding the utility of these techniques in unraveling philometrid life cycles and, in particular, to determine putative paratenic host species. Our focus was to study three species of philometrids commonly found in the estuaries of South Carolina: Philometroides paralichthydis and Philometra overstreeti from the southern flounder, Paralichthys lethostigma and Philometra carolinensis from the spotted seatrout, Cynoscion nebulosus. A total of 473 fish in 19 species known to be potential prey of the spotted seatrout and the southern flounder were dissected. Of all nematode larvae found in the mesenteries of 53 fish of 10 species, 21 specimens were determined to be philometrids using PCR. The use of PCR-RFLP allowed the identification of larvae of P. carolinensis and P. overstreeti in the freshwater goby, Ctenogobius shufeldti, and P. overstreeti in mummichog, Fundulus heteroclitus. However, 12 RFLP profiles could not be matched to control species, thus demonstrating the limitation of this technique in areas where diversity of philometrids is not well known and higher than anticipated. Similarly, COI procedures provided unknown sequences that did not match those of nine philometrid species used as controls. We concluded that although both techniques showed some usefulness and promise, at this point, however, they demonstrate the need of increasing our knowledge of marine and estuarine philometrid biodiversity.

Seasonal dynamics of Skrjabinoptera phrynosoma (Nematoda) infection in horned lizards from the Alvord Basin: temporal components of a unique life cycle.

The nematode Skrjabinoptera phrynosoma is a stomach parasite of horned lizards in the genus Phrynosoma. This nematode demonstrates a distinctive life cycle wherein entire gravid females harboring infective eggs exit with lizard feces. Pogonomyrmex spp. harvester ants collect these females and feed them to their larvae, which are the only stages of the intermediate host that can become infected. We hypothesized that the seasonal dynamics of nematode abundance within lizard hosts would be correlated with the seasonal availability of suitable intermediate hosts. To describe seasonal variation of nematode population variables and elucidate the timing of critical events in the parasite life cycle, nematodes were collected from both hosts across three collection periods in the ant-and-lizard activity season of 2008 in the Alvord Basin of southeastern Oregon. Among 3 collection periods, and across the activity season, nematodes were harvested from individual Phrynosoma platyrhinos , and the distribution of developmental categories and body lengths of nematodes was analyzed to determine the seasonal change in nematode population composition. Pogonomyrmex spp. ants were collected in pit-fall traps and dissected to determine infection prevalence. The abundance of non-gravid female and juvenile nematodes collected from lizards' stomachs decreased significantly between the early and late collection period, which was likely a consequence of the sequential conversion of these developmental categories to gravid females. The presence of gravid female nematodes peaked in cloacal and fecal collections during mid-season. The body lengths of male nematodes increased as the activity season progressed, perhaps due to growth, but their abundance remained the same. Smaller juvenile nematodes were present in late-season collections from lizards, possibly indicating new acquisitions from infected ants. We propose that once a set population of male nematodes establishes in lizards' stomachs, newly acquired juvenile nematodes develop into non-gravid females that mate, become gravid females, and exit the lizard mid-season. We additionally suggest that the exit of females may be timed with the peak foraging activity of ant intermediate hosts and access to larval ants in the nests. Infection prevalence in the intermediate host was low, with only 1 of 6,000 dissected harvester ants containing a single larval nematode. The temporal dynamics of S. phrynosoma populations within P. platyrhinos at this northern locale is most likely driven by the seasonal availability of harvester ant intermediate hosts.

New morphological data on the first-stage larvae of two Procamallanus species (Nematoda: Camallanidae) based on SEM studies.

First-stage larvae of camallanid nematodes Procamallanus (Procamallanus) laeviconchus (Wedl, 1862) and Procamallanus (Procamallanus) sp. from naturally infected Distichodus niloticus (Hasselquist) and Clarias gariepinus (Burchell), respectively, from Lake Turkana, Kenya (new geographical records) are described, being for the first time studied by scanning electron microscopy. Larvae of both species are characterised by the presence of a dorsal cephalic tooth, four submedian cephalic papillae and a pair of amphids, and by the elongate tail with several terminal digit-like processes. The latter formations probably serve for the attachment of larvae to the substrate in water when the larvae attract copepod intermediate hosts by their movements; these structures, especially their numbers, may be of taxonomic importance in camallanid nematodes.

Quantitative assessment of the nematode fauna present on Geotrupes dung beetles reveals species-rich communities with a heterogeneous distribution.

Pristionchus spp. nematodes exhibit several traits that might serve as pre-adaptations to parasitism. Under harsh environmental conditions, these nematodes can arrest development and form dauer larvae. In addition, they have been shown to live in necromenic association with a range of beetles, including dung beetles ( Geotrupes stercorosus ) on which, for example, Pristionchus entomophagus is commonly found. It has been argued that the formation of dauer larvae and the association with invertebrates represent intermediate steps towards parasitism. To better understand necromenic associations, and to gain information on Pristionchus spp. abundance and the general species composition on dung beetles, we extracted all the nematode fauna present on 114 individuals of G. stercorosus. By direct sequencing using the 18S SSU, we provide a barcode for all nematodes isolated from the beetle samples. In total, 5,002 dauer-stage nematodes were sequenced, which included Pristionchus spp., Koerneria spp. (Diplogastridae), Pelodera spp. (Rhabditidae), and Strongyloidea as well as Spirurida. Intensities of infection varied from over 1,000 nematodes isolated from a single G. stercorosus to none, with Pelodera spp. being the most abundant group isolated. This study presents the first quantitative data on the Pristionchus spp. infection of beetles.

Vaccination of eels (Anguilla japonica and Anguilla anguilla) against Anguillicola crassus with irradiated L3.

The original host of the swimbladder nematode Anguillicola crassus, the Japanese eel (Anguilla japonica) and the recently colonized European eel (Anguilla anguilla) were immunized with 40 irradiated (500 Gy) 3rd-stage larvae (L3) of this parasite and challenged with an infection of 40 normal L3. The immunization induced a significant reduction of the number of adult worms developing from the challenge infection in A. japonica, but not in A. anguilla. The induced resistance (calculated using the relation of the number of adult worms in immunized eels and in non-immunized control eels) in A. japonica was 87.3%+/-30.4%. Following a single infection, the percentage of adult worms found in A. japonica was lower as compared to A. anguilla, and the few adult worms were much smaller, revealing a lower susceptibility of A. japonica to A. crassus in comparison to A. anguilla. Both eel species developed an antibody response against A. crassus, but the level of antibody responses was not positively correlated with the protection against infection, suggesting that the antibody response is not a key element in resistance of eels against A. crassus. This study suggests that the original host of A. crassus is able to mount efficient protective immune responses against its parasite, whereas the newly acquired host seems to lack this ability.

Population dynamics and maturation cycle of Camallanus cotti (Nematoda: Camallanidae) in the Chinese hooksnout carp Opsariichthys bidens (Osteichthyes: Cyprinidae) from a reservoir in China.

The seasonal population dynamics and maturation cycle of the nematode Camallanus cotti in the posterior intestine of Chinese hooksnout carp Opsariichthys bidens have been studied in the Danjiangkou Reservoir of the Hubei Province in central China from September 2004 to November 2005. The overall prevalence, mean abundance and intensity of C. cotti among fish sampled (n=700 fish) were 47%, 2.29+/-12.38 (+/-S.D.) and 1-307 (average 4.89+/-17.74), respectively. The overall sexual ratio of female to male nematodes (excluding L3 and L4 juveniles) was 1.17:1. Statistical results showed weakly positive correlations between fish length and the number of nematodes per host. The dynamics of infection of the nematode exhibited significant seasonal pattern in changes in mean abundance. A similar pattern was found for changes in nematode prevalence, although this was not statistically significant. Higher levels of infection were observed among fish sampled in summer months and the lower in the winter. Neither the prevalence nor the abundance of the parasite was significantly different between male and female hosts. The pattern of frequency distribution of the parasite in the host was found to be over-dispersed throughout the sampling period. In addition, studies on the development and maturation of the parasite in O. bidens revealed that development (maturation), recruitment of the next generation, and reproduction may be continuous year-round, although reproduction may peak during the winter.

Some aspects of the taxonomy and biology of adult spirurine nematodes parasitic in fishes: a review.

About 300 species belonging to four superfamilies (Gnathostomatoidea, Habronematoidea, Physalopteroidea and Thelazioidea) of the nematode suborder Spirurina are known as the adult parasites of freshwater, brackish-water and marine fishes. They are placed in four families, of which the Gnathostomatidae, including Echinocephalus with a few species and the monotypic Metaleptus, are parasites of elasmobranchs, whereas Ancyracanthus contains one species in teleosts; the Physalopteridae is represented in fish by four genera, Bulbocephalus, Heliconema, Paraleptus and Proleptus, each with several species in both elasmobranchs and teleosts. The majority of fish spirurines belongs to the Rhabdochonidae, which includes 10 genera (Beaninema, Fellicola, Hepatinema, Heptochona, Johnstonmawsonia, Megachona, Pancreatonema, Prosungulonema, Rhabdochona and Vasorhabdochona) of species parasitizing mainly teleosts, rarely elasmobranchs, and the Cystidicolidae with about 23 genera (Ascarophis, Caballeronema, Capillospirura, Comephoronema, Crenatobronema, Cristitectus, Ctenascarophis, Cyclozone, Cystidicola, Cystidicoloides, Johnstonmawsonoides, Metabronema, Moravecnema, Neoascarophis, Parascarophis, Prospinitectus, Pseudascarophis, Pseudoproleptus, Salvelinema, Similascarophis, Spinitectoides, Spinitectus, Sterliadochona), with many species parasitic in teleosts only. Because of difficulties in studying fish spirurines, associated with their morphological and biological peculiarities, most species of these parasites are poorly known. It is apparent that their present classification system does not reflect phylogenetic relationships and a taxonomic revision of this nematode group, based on detailed morphological (including SEM and TEM), life history and molecular studies of individual species, is quite necessary. In Cystidicolidae, several genera have been based on details in the cephalic structures visible only with the aid of SEM, but it will be evident whether or not these tiny features are of generic importance only when more cystidicolids are described using SEM and comparative molecular data become available. Data on the biology of fish spirurines are scarce. In known cases, their life cycles involve aquatic arthropods (crustaceans or insects) as intermediate hosts, in which, sometimes, the larvae undergo a precocious development and may even attain adulthood and become gravid in these invertebrates; sometimes, fish paratenic hosts are known to occur in cystidicolids parasitizing as adults piscivorous definitive hosts. Some spirurine species are pathogenic and are known as causative agents of serious fish diseases. Consequently, further detailed studies on fish spirurines are significant not only from the theoretical viewpoint, but they may also have practical implications.

Comparative studies on intestine ultrastructure of third-stage larvae and adults of Cystidicoloides ephemeridarum (Nematoda, Cystidicolidae).

The intestinal epithelium of third-stage larvae and adults of Cystidicoloides ephemeridarum from haemocoel of mayflies and stomach of brown trout was studied by electron microscopy and cytochemistry. In section, the intestine of both stages is composed of a single layer of about ten undifferentiated intestinal cells in a ring. A labyrinth of deep invaginations is present in the basal region of each cell. The apical surface is modified into well developed, regularly arranged microvilli. These, together with numerous organelles engaged in metabolism and a well defined gut lumen filled with unidentifiable material suggest that the intestine may function in digestion and absorption during both stages. The adults seem to feed upon the semifluid content of the stomach of brown trout. Fortuitous oral infection with undetermined bacteria in vitro led to degenerative changes in the intestinal tissue and probably caused death of the infected specimens. Up to 75% of the cell volume in the L(3) is occupied by glycogen deposits. In the adults, a minor portion of glycogen, together with lipid droplets, has been observed. The adults are considered to rely more on aerobic metabolism, whereas anaerobic metabolism (glycolysis) may prevail in L(3).

Ultrastructure of the body wall of infective larvae of Cystidicoloides ephemeridarum (Nematoda, Cystidicolidae) from mayflies.

Scanning and transmission electron microscopic examinations of tissue-dwelling third-stage larvae of the nematode Cystidicoloides ephemeridarum from the intermediate host (Ephemera danica) were carried out with respect to the morphological changes in the body wall associated with the transfer from the intermediate to the definitive host. The ultrastructure of the cephalic end and zonation of the somatic cuticle of infective larvae basically correspond with those of adults. The somatic cuticle is composed of a fuzzy epicuticle, an outer and inner cortical zone, a median zone and a basal zone consisting of three subzones. Globular bodies are absent from the median zone of the infective larvae. The lateral hypodermal cords of the infective larvae are cellular, consisting of a median cell enclosed by two sublateral cells. The excretory canal is present within each of the lateral cord in both the infective and adult stages. Walls of the excretory canals contain Golgi-derived vesicles which communicate with the lumen of the canal. Large deposits of glycogen, suggesting anaerobic respiration, are present in the hypodermal cords and noncontractile parts of the muscle cells of the third-stage larvae within the intermediate host.

Larval spirurida (Nematoda) from the crab Macrophthalmus hirtipes in New Zealand.

Previously undescribed third-stage larvae of two species of Spirurida were found in the haemocoel of the stalk-eyed mud crab Macrophthalmus hirtipes (Heller) (Ocypodidae) in New Zealand. Examinations by light and scanning electron microscopy showed that the larger larvae (about 7 mm long) belonged to a species of Ascarophis van Beneden, 1871 (Cystidicolidae), the genus including parasites of fishes, whereas the smaller larvae (about 4-5 mm long) belonged to the Acuariidae, a family with species parasitic as adults mostly in aquatic birds. In a sample of 82 specimens of M. hirtipes collected in July 2002 from Papanui Inlet, on Otago Peninsula, South Island, 74 crabs (90.2%) were infected with larval nematodes with an intensity of 1-18 (mean 4.6) nematodes per crab; no distinction between nematode species was made in these estimates, although juvenile Acuariidae greatly outnumbered larval Ascarophis. Apparently, crabs play a role as intermediate hosts of these nematode species. This is the first record of larval representatives of Cystidicolidae and Acuariidae from invertebrates in the Australasian Region.

The development and morphogenesis of Camallanus cotti Fujita, 1927 (Nematoda: Camallanidae), with notes on its phylogeny and definitive host range.

The freshwater fish nematode Camallanus cotti Fujita, 1927 (Spirurida: Camallanidae) is naturally distributed in East, South and Southeast Asia. During the past three or four decades the species has been disseminated to Europe, North America, Australia and Hawaii, mainly due to the extensive ornamental fish trade. In the present study, its development and larval morphogenesis is described. Newborn first-stage larvae are ingested by copepods and within hours reach the haemocoel, where the worms moult twice. At 22 degrees C ambient temperature, fully-developed third-stage larvae appear on day 11 p.i. After transmission into the fish intestine two further moults occur before the adult stage is reached. At 23 degrees C water temperature, the final moult is initiated around day 33 p.i. in males and between 34 and 42 days p.i. in females. Morphologically the young and older adults are similar, but the buccal capsule of both young males and young females is not yet fully sclerotised. Based on the close similarity in the development and adult morphology of the buccal capsule in C. cotti and three congeners from Europe and North America, we suggest, in general agreement with earlier workers, that the Camallanus Railliet & Henry, 1915 originated in tropical Asia and subsequently radiated to species when adapting to new climatic and/or host-related conditions in the Old World and the New World. C. cotti may thus represent a lineage of speciation which did not apparently involve extensive changes in relation to their natural geographical distribution or definitive host range.