Morphological and molecular genetic diversity of Strongyluris calotis (Nematoda: Ascaridida: Heterakidae) in South East and East Asian lizards.

Strongyluris calotis is a heterakid nematode in the large intestine of agamid lizards (Reptilia: Sauria: Agamidae) from the Oriental Region. The standard light microscopic definition of the species counts the "caudal papillae" as 10 pairs on male worms. However, previous work from our group using scanning electron microscopy (SEM) on the heterakid from agamid lizards in Japan, Taiwan, and Singapore revealed that this counting contained a pair of phasmids and that two pairs of postcloacal papillae were completely fused to form a pair of united papillae, thus resulting in "10 pairs." In the present study, we examined S. calotis specimens from the Emma Gray's forest lizard, Calotes emma (Agamidae), living in the plain forest at low altitude, and the Vietnam false bloodsucker, Pseudocalotes brevipes (Agamidae), living in the mountainous forest at high altitude in the northern part of Vietnam. Using SEM, the arrangement of caudal papillae in male worms from an Emma Gray's forest lizard was found to be comparable to classical S. calotis specimens from agamid lizards collected in Japan, Taiwan, and Singapore. However, male worms from Vietnam false bloodsuckers did not have a pair of united papillae but had 10 pairs of independent caudal papillae with a pair of phasmids. Molecular genetic analyses of the ribosomal RNA gene (rDNA) of worms of the classical S. calotis morphotype from Japan and Singapore and two S. calotis morphotypes from Vietnam demonstrated absolutely identical nucleotide sequences of partial 18S rDNA (at least 1764 base pairs (bp)) and 5.8S rDNA (158 bp). However, intraspecific differences were detected in other regions of the rDNA, related to the geographical distribution of hosts regardless of morphotype: 97.8-98.5 % identity (443-446 bp/453 bp) in the internal transcribed spacer (ITS)-1 region, 96.6-98.0 % identity (425-431 bp/440 bp) in the ITS-2 region, and 99.6-99.7 % identity (1149-1151 bp/1154 bp) in the 28S rDNA. Thus, in the future, taxonomic relationships of S. calotis distributed widely in the Oriental Region as well as other nominal Oriental Strongyluris spp., currently six in number, need to be extensively explored based on molecular genetic analyses in addition to intensive morphological characterization.

A new Cosmocercoides species (Nematoda: Cosmocercidae), C. tonkinensis n. sp., in the scale-bellied tree lizard (Acanthosaura lepidogaster) from Vietnam.

A new cosmocercid nematode species, Cosmocercoides tonkinensis n. sp., is described from the scale-bellied tree lizard (Acanthosaura lepidogaster) in the northern and central parts of Vietnam. The new species is characterized by medium-sized male worms (4.2-5.1 mm in length and 0.34-0.37 mm in width) relative to known members of the genus, with lateral alae, two sharply pointed spicules of equal length (0.22-0.26 mm in length), a gubernaculum (0.113-0.122 mm in length), 16 or 17 pairs of caudal rosettes, and the presence of somatic papillae. Female worms are slightly larger than male worms (5.3-5.5 mm in length and 0.32-0.42 mm in width), with the vulva situated at 3/5 from the anterior end, and elliptical embryonated eggs, 0.064- 0.084 mm long by 0.040-0.048 mm wide. From 19 recorded species of the genus, the morphology of C. tonkinensis n. sp. is closest to C. multipapillata, C. bufonis, and C. pulcher reported from toads and frogs in East Asia. The present new species is differentiated from them by the number of caudal rosettes, tail length relative to body length, presence of somatic papillae and lateral alae, and embryonated eggs. Furthermore, after C. variabilis in North America and C. sauria in Brazil, this new species is only the third species to be recorded from a reptilian host. The 18S ribosomal RNA gene (rDNA) of the new species is almost identical to that of C. dukae infecting land snails and slugs in North America. Between the present new species and C. pulcher from a toad (Bufo japonicus) in Japan, remarkably fewer nucleotide changes were noticed in the 18S to 28S rDNA including the internal transcribed spacer regions. The molecular phylogenetic position of the genus Cosmocercoides is briefly discussed.