Pseudoacanthocephalus toshimai sp. nov. (Palaeacanthocephala: Echinorhynchidae), a common acanthocephalan of anuran and urodelan amphibians in Hokkaido,Japan, with a finding of its intermediate host

The Ezo brown frog (Rana pirica) and the Ezo salamander (Hynobius retardatus) are endemic species of Hokkaido, the northernmost island of Japan. Intestinal adult acanthocephalans are common in these amphibians. A molecular identification based on nuclear and mitochondrial DNA markers demonstrated that the parasites from the anuran and the urodelan are the same species. In the neighboring Honshu island, another acanthocephalan from ranid frogs (e.g. Rana japonica and Rana ornativentris) has been identified as Acanthocephalus lucidus. The counterpart species from the amphibians of Hokkaido was morphologically indistinguishable from A. lucidus. However, clear genetic distinctiveness between the two allopatric populations (separated by islands) indicated the entity of a cryptic species. A phylogenetic tree inferred from sequences of 28S ribosomal DNA showed that the acanthocephalans from Honshu and Hokkaido belong to the genus Pseudoacanthocephalus. Therefore, Pseudoacanthocephalus toshimai sp. nov. is proposed for the cryptic species in Hokkaido, together with the transfer of A. lucidus in Honshu to Pseudoacanthocephalus lucidus comb. nov. The present field survey further demonstrated Ligidium japonicum, an isopod crustacean living in the litter layer of forests, to be an intermediate host of the new species.     

New and already known acanthocephalans from amphibians and reptiles in Vietnam, with keys to species of Pseudoacanthocephalus Petrochenko, 1956 (Echinorhynchidae) and Sphaerechinorhynchus Johnston and Deland, 1929 (Plagiorhynchidae)

Adults of 2 new species in 2 orders of acanthocephalans obtained from the intestines of terrestrial amphibians and reptiles collected between 1998 and 2004 in Vietnam are described here. Pseudoacanthocephalus nguyenthileae n. sp. (Palaeacnthocephala: Echinorhynchidae) was collected from 5 species of terrestrial amphibians: (1) the common Sunda toad Bufo melanostictus Schneider (Bufonidae); (2) Paa verucospinosa (Bourret); (3) Gunther's Amoy frog Rana guentheri Boulenger; (4) Taipei frog R. taipehensis Denburgh (Ranidae), and (5) the Burmese whipping frog Polypedates mutus (Smith) (Racophoridae); as well as from the Chinese cobra Naja atra Cantor (Reptilia: Elapidae) and house gecko Hemidactylus frenatus Dumeril and Bibron (Reptilia: Gekkonidae). Sphaerechinorhynchus maximesospinus n. sp. (Plagiorhynchidae: Sphaerechinorhynchinae) was isolated from a king cobra Ophiophagus hannah (cantor) (Reptilia: Elapidae). Cystacanths of Porrorchis houdemeri (Joyeux and Baer, 1935) Schmidt and Kuntz, 1967 (Plagiorhynchidae: Porrorchinae) obtained from the mesenteries of banded krait Bungarus fasciatus (Schneider) (Reptilia: Elapidae), a paratenic host, are reported for the first time. Keys to the species of Pseudoacanthocephalus and Sphaerechinorhynchus are included. Characteristic features distinguishing the new species from related taxa include: P. nguyenthileae has 15-19 (usually 16-18) proboscis hook rows, each with 5-6 hooks that progressively increase in length and size posteriorly. The largest, intermediate, and smallest proboscis hooks of S. maximesospinus are the middle, anterior, and posterior hooks, respectively; the proboscis and neck are enclosed in a membrane. Morphometric characteristics of P. nguyenthileae show host-related variability.     

Molecular insight into systematics,host associations,life cycles and geographic distribution of the nematode family Rhabdiasidae

Rhabdiasidae Railliet, 1915 is a globally distributed group of up to 100 known species of nematodes parasitic in amphibians and reptiles. This work presents the results of a molecular phylogenetic analysis of 36 species of Rhabdiasidae from reptiles and amphibians from six continents. New DNA sequences encompassing partial 18S rDNA, ITS1, 5.8S rDNA, ITS2 and partial 28S rDNA regions of nuclear ribosomal DNA were obtained from 27 species and pre-existing sequences for nine species were incorporated. The broad taxonomic, host and geographical coverage of the specimens allowed us to address long-standing questions in rhabdiasid systematics, evolution, geographic distribution, and patterns of host association. Our analysis demonstrated that rhabdiasids parasitic in snakes are an independent genus sister to the rest of the Rhabdiasidae, a status supported by life cycle data. Based on the combined evidence of molecular phylogeny, morphology and life cycle characteristics, a new genus Serpentirhabdias gen. nov. with the type species Serpentirhabdias elaphe (Sharpilo, 1976) comb. nov. is established. The phylogeny supports the monophyly of Entomelas Travassos, 1930, Pneumonema Johnston, 1916 and the largest genus of the family, Rhabdias Stiles and Hassall, 1905. DNA sequence comparisons demonstrate the presence of more than one species in the previously monotypic Pneumonema from Australian scincid lizards. The distribution of some morphological characters in the genus Rhabdias shows little consistency within the phylogenetic tree topology, in particular the apical structures widely used in rhabdiasid systematics. Our data suggest that some of the characters, while valuable for species differentiation, are not appropriate for differentiation among higher taxa and are of limited phylogenetic utility. Rhabdias is the only genus with a cosmopolitan distribution, but some of the lineages within Rhabdias are distributed on a single continent or a group of adjacent zoogeographical regions. Serpentirhabdias, Entomelas and Pneumonema show rather strict specificity to their host groups. The evolution of the Rhabdiasidae clearly included multiple host switching events among different orders and families of amphibians as well as switching between amphibians and squamatan reptiles. Only a few smaller lineages of Rhabdias demonstrate relatively strict associations with a certain group of hosts.     

Acanthocephalans from some frogs and toads (Anura) and chameleons (Squamata) from Tanzania with the description of a new species

The following Acanthocephala, all Echinorhynchidae, were found on examination of 229 amphibians and 191 reptiles collected in the Eastern Arc Range, Tanzania: Acanthocephalus bufonis from the frogs Arthroleptis stenodactylus and Arthroleptides martiensseni, the toad Nectophnyroides viviparous, and the chameleon Rhampholeon brevicaudatus; Pseudoacanthocephalus betsileo from the frogs Probreviceps macrodactylus and Ptychadero anchietae; and Pseudoacanthocephalus rhampholeontos n. sp. from the chameleons Rhampholeon sp. and Rhampholeon uluguruensis. Pseudoacanthocephalus rhampholeontos differs from the only other acanthocephalan occurring in chameleons, Pseudoacanthocephalus bigueti, in being a larger worm with fewer, smaller hooks in the proboscis armature. The status of the Acanthocephalus and Pseudoacanthocephalus is still problematic and requires further attention and study.     

Acanthocephala in amphibians (Anura) and reptiles (Squamata) from Brazil and Paraguay with description of a new species

In a survey of 1,732 amphibians and reptiles collected across S?o Paulo Province, Brazil, and 7 provinces in Paraguay, 26 species were found infected with acanthocephalans. Of 1,510 anurans, 14 anurans, representing 11 species, were infected with cystacanths of Centrorhynchus spp. and 1 anuran with cystacanths of Oligacanthorhynchus sp. Of 107 lizards, 1 lizard was infected with cystacanths of Centrorhynchus sp. and 1 lizard with cystacanths of Oligacanthorhynchus sp. Acanthocephalus caspanensis was found in 3 anurans (3 species) and Acanthocephalus lutzi in 3 anurans (2 species) and 2 snakes (2 species). The systematic position of A. lutzi cannot be resolved using presently available morphological data. Acanthocephalus saopaulensis n. sp. was found in a single individual of Bufo ictericus. The new species can be differentiated from all its congeners except A. caspanensis in having a sigmoid-shaped male posterior end and from A. caspanensis in having a proboscis armature of 16 rows of 5-7 hooks rather than 18-19 rows of 6-7 hooks and larger eggs. The status of Acanthocephalus and Pseudoacanthocephalus continues to be problematic.     

Molecular data reveal a new species of Seuratascaris Sprent, 1985 (Nematoda: Ascaridoidea) from Quasipaa exilispinosa (Liu & Hu) (Amphibia: Anura)

The genus Seuratascaris Sprent, 1985 is a group of obligate nematode parasites of amphibians. In the present study, a new species of Seuratascaris, S. physalis sp. n. was described using light and scanning electron microscopy based on specimens collected from Quasipaa exilispinosa (Liu & Hu) (Amphibia: Anura) in China. The new species differs from S. numidica (Seurat, 1917) by the cuticle of the cervical region distinctly inflated to form a cephalic vesicle-like structure and the absence of single medio-ventral precloacal papilla. The molecular characterization of the nuclear large ribosomal DNA (28S) and internal transcribed spacer (ITS) and the mitochondrial cytochrome c oxidase subunit 1 (cox1), cytochrome c oxidase subunit 2 (cox2) and 12S small subunit ribosomal RNA gene of S. physalis sp. n., together with the 28S, cox2 and 12S of S. numidica are provided for the first time. Molecular analysis revealed the presence of high level of interspecific genetic variation between the two species in the ITS (5.50%), cox1 (13.3%), cox2 (10.6%) and 12S regions (10.5%), which strongly supported that S. physalis sp. n. represented a different species from S. numidica. Angusticaecum ranae Wang, Zhao & Chen, 1978 reported from the frog Quasipaa spinosa (David) (Anura: Dicroglossidae) in China was transferred into the genus Seuratascaris as S. ranae (Wang, Zhao & Chen, 1978) comb. n. based on the morphology of lips and the presence of very short and robust spicules without alae and small numbers of precloacal papillae. The present study provided useful genetic data for molecular identification of species of Seuratascaris and provides the foundation for being able to determine if S. numidica represents a species complex of some sibling species or a single species.     

Icosiella turgeocauda n. sp. (Nematoda: Onchocercidae) and Seuratascaris numidica (Nematoda: Ascarididae), parasites of the frog,Rana cancrivora (Anura: Ranidae), from Luzon,Republic of the Philippines

Icosiella turgeocauda n. sp. from the intestinal mesenteries of Rana cancrivora collected at Luzon, Republic of the Philippines, is described and illustrated. Icosiella turgeocauda n. sp. represents the ninth species to be assigned to the genus and is easily differentiated from all the previously described species by the position of the vulva and the presence of bilateral umbos on the caudal end of the male. Seuratascaris numidica also was found. The Philippines represents a new location record for S. numidica.     

湖北神农架国家级自然保护区两栖爬行动物物种多样性

对神农架自然保护区两栖爬行动物进行调查,整理了神农架自然保护区两栖爬行动物名录共71种,新发现湖北省新分布种6种,其中腾格里蛙Rana tenggerensis Zhao,Macey et Papenfuss、宁陕小头蛇 Oligodon ningshaanensis Yuan离原分布区较远,神农架新分布种3种,神农架两栖爬行动物的区系特征以东洋界(74.69%)为主,广布种(18.31%)、古北界(7%)种类次之,体现了南北混杂的特点.随着海拔的升高,两栖爬行动物生物多样性总体呈下降趋势,400～1600 m两栖爬行动物生物多样性最高,随海拔下降的趋势不明显,1900 m以上多样性急剧下降.     

Sequencing and analysis of the internal transcribed spacers (ITSs) and coding regions in the EcoR I fragment of the ribosomal DNA of the Japanese pond frog Rana nigromaculata

The rDNA of eukaryotic organisms is transcribed as the 40S-45S rRNA precursor, and this precursor contains the following segments: 5' - ETS - 18S rRNA - ITS 1 - 5.8S rRNA - ITS 2 - 28S rRNA - 3'. In amphibians, the nucleotide sequences of the rRNA precursor have been completely determined in only two species of Xenopus. In the other amphibian species investigated so far, only the short nucleotide sequences of some rDNA fragments have been reported. We obtained a genomic clone containing the rDNA precursor from the Japanese pond frog Rana nigromaculata and analyzed its nucleotide sequence. The cloned genomic fragment was 4,806 bp long and included the 3'-terminus of 18S rRNA, ITS 1, 5.8S rRNA, ITS 2, and a long portion of 28S rRNA. A comparison of nucleotide sequences among Rana, the two species of Xenopus, and human revealed the following: (1) The 3'-terminus of 18S rRNA and the complete 5.8S rRNA were highly conserved among these four taxa. (2) The regions corresponding to the stem and loop of the secondary structure in 28S rRNA were conserved between Xenopus and Rana, but the rate of substitutions in the loop was higher than that in the stem. Many of the human loop regions had large insertions not seen in amphibians. (3) Two ITS regions had highly diverged sequences that made it difficult to compare the sequences not only between human and frogs, but also between Xenopus and Rana. (4) The short tracts in the ITS regions were strictly conserved between the two Xenopus species, and there was a corresponding sequence for Rana. Our data on the nucleotide sequence of the rRNA precursor from the Japanese pond frog Rana nigromaculata were used to examine the potential usefulness of the rRNA genes and ITS regions for evolutionary studies on frogs, because the rRNA precursor contains both highly conserved regions and rapidly evolving regions.     

First steps to understand the systematics of Echinorhynchidae Cobbold, 1876 (Acanthocephala), inferred through nuclear gene sequences

Acanthocephalans of the order Echinorhynchida are one of the most diverse groups in their phylum, with approximately 470 species classified into 11 families that largely consist of parasites of freshwater, brackish and marine fishes and, sporadically, reptiles and amphibians distributed worldwide. Previous phylogenies inferred with molecular data have supported the paraphyly or polyphyly of some families, suggesting that most of them have been diagnosed based on unique combinations of characters, rather than shared derivative features. We expand the taxonomic sampling of several genera such as Acanthocephalus, Echinorhynchus and Pseudoacanthocephalus of Echinorhynchidae from diverse biogeographical zones in the Americas, Europe and Asia with the aim of testing the monophyly of the family by using two molecular markers. Sequences from small (SSU) and large (LSU) subunits of ribosomal DNA were obtained for six species representing the genera Acanthocephalus and Echinorhynchus from the Neotropical, Nearctic, Palearctic and Oriental regions. These sequences were aligned with other sequences available in the GenBank dataset from Echinorhynchidae. Phylogenetic trees inferred with the combined (SSU + LSU) and the individual data sets consistently placed the genera Acanthocephalus, Pseudoacanthocephalus and Echinorhynchus into three independent lineages. Two families, Paracanthocephalidae Golvan, 1960, and Pseudoacanthocephalidae Petrochenko, 1956, were resurrected to accommodate the genera Acanthocephalus and Pseudoacanthocephalus, respectively. The species of the genus Acanthocephalus from the Nearctic, Palearctic and Oriental biogeographic regions formed a clade that was well supported. However, Acanthocephalus amini from the Neotropical region was nested inside Arhythmacanthidae. Therefore, the genus Calakmulrhynchus was created to accommodate A. amini and resolve the paraphyly of Acanthocephalus. Finally, the diagnoses of the families Echinorhynchidae and Arhythmacanthidae were amended. The molecular phylogenies should be used as a taxonomic framework to find shared derived characters (synapomorphies) and build a more robust classification scheme that reflects the evolutionary history of the acanthocephalans.     

Amphibians and reptiles from the Early Miocene of the Bardenas Reales of Navarre (Ebro Basin, Iberian Peninsula)

The Lower Miocene deposits of the Bardenas Reales of Navarre (NW Ebro Basin, northern Iberian Peninsula) have yielded a diverse vertebrate fauna, including remains of amphibians and reptiles. These remains occur in several localities in the Tudela Formation. The fossiliferous levels belong to the Biozones MN2b-3 (Biozones Z-A of the Ramblian, i.e., Late Aquitanian to Early Burdigalian in age). The amphibians and reptiles represent at least 13 out of 37 vertebrate species. Amphibians consist of a salamandrid urodele and two or three anurans. All the turtles are cryptodirans and consist of the chelydrid Chelydropsis apellanizi, the testudinids Ptychogaster (Temnoclemmys) bardenensis and Ptychogaster ronheimensis, and a Trionychinae indet. Squamates are represented by the anguid lizard Ophisaurus sp., a non-anguid lacertilian, an amphisbaenian, the erycine boid? Eryx sp., and indeterminate colubrids. Crocodilian remains are assigned to the basal alligatoroid Diplocynodon sp. The fossil associations of the Bardenas Reales of Navarre suggest that the vertebrates lived in the centre of an endoreic basin with stretches of water under intertropical to subtropical climatic conditions.     

Frequent infections of mountain stream fish with the amphibian acanthocephalan,Pseudoacanthocephalus toshimai (Acanthocephala: Echinorhynchidae)

Pseudoacanthocephalus toshimai is an intestinal acanthocephalan parasite of amphibians in Hokkaido, the northernmost island of Japan. In this study, common freshwater fish of the families Salmonidae and Cottidae in mountain streams around the Kamikawa basin of Hokkaido were examined for acanthocephalan infections with P. toshimai. A total of 160 salmonids and 14 cottids were caught in 4 streams by bait fishing during summer and autumn seasons of 2019. Adult acanthocephalans were found only from the salmonids, namely, Salvelinus leucomaenis leucomaenis, Salvelinus malma krascheninnikovi, Oncorhynchus masou, and Oncorhynchus mykiss. The maximum prevalence reached 58.1% in S. leucomaenis, but the mean worm burden was at low levels (e.g., 3.1 in S. leucomaenis and 2.2 in S. malma). All of the acanthocephalans were identified to P. toshimai by morphological observation and DNA barcoding. Although the male acanthocephalans became sexually mature, the females never reached the gravid adult stage, suggesting that salmonids are unsuitable or aberrant hosts for P. toshimai. The infected fish were found exclusively from a small stream with bush, in which a large habitat of amphibians is included. Ligidium japonicum, a terrestrial isopod, collected from the habitat was highly infected with cystacanth larvae of P. toshimai. The observation of fish stomach contents directly demonstrated that small salmonids eat L. japonicum. The terrestrial isopods, which are washed away by rain into a stream, seem to be a source of salmonid infections with P. toshimai. The habitat of intermediate hosts should be emphasized in the taxonomy of the closely related genera Acanthocephalus and Pseudoacanthocephalus.     

Additional Records of Trematodes of Amphibians and Reptiles from Taiwan

Three monogenetic and 23 digenetic trematodes of amphibians and reptiles are reported from Taiwan, 14 for the first time. One new family and genus and five new species of Digenea are described: Omphalometridae, Leurosoma taiwanense sp.n. from Bufo bufo bankorensis; Dicrocoeliidae, Paradistomum taiwanense sp.n. from Natrix stolata; Ocadiatrematidae, Ocadiatrema taiwanense fam. gen. et sp.n. from Ocadia sinensis; Lecithodendriidae, Prosthodendrium (Paralecithodendrium)taiwanense sp.n. from Ocadia sinensis; Pleurogenidae, Nenimandijea taiwanense sp.n. from Rana plancyi fukienensis .     

Two new Neuratelia Rondani (Diptera,Mycetophilidae) species from Western Palaearctic: a case of limited congruence between morphology and DNA sequence data

Two new Mycetophilidae species, Neuratelia jabalmoussae sp. n. and Neuratelia salmelai sp. n. are described on the basis of material collected from Lebanon, Estonia and Finland. Detailed figures of male terminalia and photographs of general facies are provided along with discussions of their morphological distinction from sibling species. For the first time molecular characters are used to distinguish new fungus gnat species. Molecular analysis relies on cytochrome oxidase subunit one (COI) but has additionally been corroborated by information from the 28S and ITS2 regions of nuclear ribosomal DNA. Situations where morphological and molecular data provide conflicting evidence for species delimitation are discussed. A new country record from Georgia is provided for Neuratelia caucasica.     

First record of Thubana species in the Philippines (Lepidoptera,Lecithoceridae), with description of two new species

Two species of the genus Thubana Walker (Lepidoptera, Lecithoceridae) are found for the first time in the Philippines and described as new: T. pedicucullata sp. nov. and T. gyrostigmatis sp. nov. Photos of adults and the genitalia for the new species are provided.     

The latest Early Pleistocene amphibians and reptiles from Kaiafas (Greece) and the first record of fossil Ophiomorus (Squamata,Scincidae)

Together with the rest of the Balkan Peninsula, Greece acted as a refuge for thermophilic amphibians and reptiles during the Quaternary Glaciations. In this work we provide new herpetological data on the latest Early Pleistocene locality of Kaiafas, in western Peloponnese. The site yielded one indeterminate salamandrid urodelan (Salamandridae indet.), one tree frog (Hyla gr. H. arborea), one indeterminate testudinid (Testudinidae indet.), at least two lacertid lizards (Lacertidae indet. A and B), one scincid lizard (Ophiomorus sp.), one anguid lizard (Pseudopus sp.), and one indeterminate “colubrine” snake (“Colubrinae” indet.). This assemblage is indicative of a surrounding environment with water bodies, wooded areas, and possibly loose soils with rocks and stones. In spite of being based on few remains, the study of the amphibians and reptiles from Kaiafas resulted in the identification of the first fossil of Ophiomorus known worldwide. Moreover, the two lacertid taxa displayed very peculiar morphological features which are either previously unknown or rare in that clade.