ABSTRACT Falcaustra vietnamensis n. sp. is described from the small intestine of Physignathus cocincinus from north-central Vietnam. The new species is characterized by the large male worms (20.2–28.8 mm in length and 557–724 μm in width) relative to known members of the genus, 2 sharply pointed alate spicules of equal length (1,128–1,256 μm in length), gubernaculum including 2 separate pieces, 1 ventral with a pointed distal end and 1 dorsal with a blunt distal end (164–192 μm and 155–172 μm in length, respectively), and 12 pairs of caudal papillae. Female worms are larger than male worms (24.2–34.1 mm in length and 532–735 μm in width), with the vulva situated in the posterior half of body, and elliptical eggs, 60–70 μm long by 42–47 μm wide. Falcaustra vietnamensis n. sp. represents the 38th species assigned to the genus and the third species recorded from a lizard host in the Oriental biogeographical region. Partial sequences of the 18S ribosomal RNA gene (rDNA), internal transcribed spacer regions ( ITS ), and cytochrome c oxidase subunit 1 ( COI ) are provided for the new species. The molecular phylogenetic position of the genus Falcaustra is briefly discussed.

Whole-length esophagi of 111 Murrah cross water buffaloes ( Bubalus bubalis ) were collected in the Kathmandu and Chitwan districts of Nepal from December 2009 to February 2010. Gullet worms showing a typical epithelium-dwelling character were detected in 13 of 53 (24.5%) buffaloes in Kathmandu and in 5 of 58 (8.6%) buffaloes in Chitwan. The worms' morphology and measurements were identical to those of Gongylonema pulchrum Molin, 1857, except for the length of the left spicules relative to the body length. Scanning electron microscopy did not detect any further morphological differences regarding the collected specimen from Nepal compared with G. pulchrum . The ribosomal RNA gene (rDNA), including internal transcribed spacer (ITS) 1 and 2, and a partial region of the cytochrome c oxidase subunit I (COI) of mitochondrial DNA of the worms were characterized and compared with those of G. pulchrum collected from cattle, deer, wild boars, and monkeys in Japan and from cattle in Iran. The 18S, 5.8S, and 28S rDNA nucleotide sequences of the buffalo-collected worms had 99.8% (1,779/1,782), 100% (158/158), and 98.3–98.8% (3,494–3,507/3,551) identities, respectively, with those of G. pulchrum from the other host mammals. The ITS regions exhibited higher variations between the buffalo-collected worms and G. pulchrum from the other host mammals (85–88% identity for ITS1 and 56–80% identity for ITS2). The COI also showed lower identities (89.2–90.2%), although only a single amino acid substitution was noted compared with the majority of G. pulchrum samples collected in Japan. Based on these molecular genetic characters in the rDNA and COI mitochondrial DNA, together with a shorter left spicule length relative to body length, the gullet worms isolated from buffaloes in Nepal might belong to a distinct local or buffalo-preferring population of G. pulchrum , although its geographical distribution on the continent and host specificity remain to be clarified.

: Skrjabinema kamosika sp. n. (Nematoda: Oxyuridae) is described based on both sexes collected from endemic Japanese serows, Capricornis crispus (Mammalia: Bovidae), of Wakayama Prefecture, western Honshu Island, and Kochi Prefecture, Shikoku Island, Japan. It is readily distinguished from the congeners by having tricuspid sub-interlabial projections in males, large triangular lateral lobes of the lips that do not reach the mouth rim, and a large length ratio of alate/nonalate portions of the tail in females. Close identity in the nucleotide arrangement of ITS2 region between the present material and that from C. crispus of Iwate Prefecture, northern Honshu Island, suggests wide distribution of S. kamosika in Japan. In partial 18S rDNA nucleotide sequence, 0.77% differed from that of Skrjabinema sp. from Capra hircus in the United States.

abstract Enterobius (Enterobius) macaci Yen, 1973 (Nematoda: Oxyuridae: Enterobiinae) was collected from a Japanese macaque, Macaca fuscata , in Nara and Yamaguchi Prefectures, Honshu Island, Japan, for the first time. A redescription is presented along with DNA sequence data. This pinworm is a typical member of the subgenus Enterobius and is characteristic in the spicule morphology, being readily distinguished from other congeners. Phylogenetic analyses based on 18S ribosomal RNA gene (rDNA) and mitochondrial DNA (mtDNA) Cox1 gene assign its position in the pinworm lineage adapted to the Old World primates, showing divergence before the splitting of the chimpanzee and human pinworms.

Peripheral blood from a ball python ( Python reginus ) imported from Ghana was cultured in Barbour-Stoenner-Kelly (BSK) medium for Borrelia spp. isolation, resulting in the prominent appearance of free, and clusters of, trypanosomes in a variety of morphological forms. The molecular phylogenetic characterization of these cultured trypanosomes, using the small subunit rDNA, indicated that this python was infected with a species closely related to Trypanosoma varani Wenyon, 1908 , originally described in the Nile monitor lizard ( Varanus niloticus ) from Sudan. Furthermore, nucleotide sequences of glycosomal glyceraldehyde-3-phosphate dehydrogenase gene of both isolates showed few differences. Giemsa-stained blood smears, prepared from the infected python 8 mo after the initial observation of trypanosomes in hemoculture, contained trypomastigotes with a broad body and a short, free flagellum; these most closely resembled the original description of T. varani , or T. voltariae Macfie, 1919 recorded in a black-necked spitting cobra ( Naja nigricollis ) from Ghana. It is highly possible that lizards and snakes could naturally share an identical trypanosome species. Alternatively, lizards and snakes in the same region might have closely related, but distinct, Trypanosoma species as a result of sympatric speciation. From multiple viewpoints, including molecular phylogenetic analyses, reappraisal of trypanosome species from a wide range of reptiles in Africa is needed to clarify the relationship of recorded species, or to unmask unrecorded species.

Neotropical primates of the Cebidae and Callitrichidae, in their natural habitats, are frequently infected with a variety of trypanosomes including Trypanosoma cruzi , which causes a serious zoonosis, Chagas' disease. The state of trypanosome infection after a 30-day quarantine period was assessed in 85 squirrel monkeys ( Saimiri sciureus ) and 15 red-handed tamarins ( Saguinus midas ), that were wild-caught and exported to Japan as companion animals or laboratory animals, for biomedical research, respectively. In addition to many microfilariae of Mansonella ( Tetrapetalonema ) mariae at a prevalence of 25.9%, and Dipetalonema caudispina at a prevalence of 3.5%, a few trypomastigotes of Trypanosoma ( Megatrypanum ) minasense were detected in Giemsa-stained thin films of blood from 20 squirrel monkeys at a prevalence of 23.5%. Although few T. minasense trypomastigotes were found in Giemsa-stained blood films from tamarins, a buffy-coat examination detected trypanosomes in 12 red-handed tamarins (80.0%), and PCR amplification of a highly variable region of the small subunit ribosomal RNA genes (SSU rDNA) for Trypanosoma spp. detected the infection in 14 of the 15 tamarins (93.3%). Nucleotide sequences of the amplicons were identical for trypanosomes from tamarins and squirrel monkeys, indicating a high prevalence but low parasitemia of T. minasense in imported Neotropical nonhuman primates. Based on the SSU rDNA and 5.8S rDNA, the molecular phylogenetic characterization of T. minasense indicated that T. minasense is closely related to trypanosomes with Trypanosoma theileri -like morphology and is distinct from Trypanosoma ( Tejeraia ) rangeli , as well as from T. cruzi . Using some blood samples from these monkeys, amplification and subsequent sequencing of the glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene fragments detected 4 trypanosome genotypes, including 2 types of T. cruzi clade, 1 type of T. rangeli clade, and 1 T. rangeli -related type, but failed to indicate its phylogenetic position based on the gGAPDH gene. Furthermore, species ordinarily classified in the Megatrypanum by morphological criteria do not form a clade in any molecular phylogenetic trees based on rDNA or gGAPDH genes.

A total of 210 individuals of 13 species belonging to 4 subfamilies of Muridae imported into Japan as pets were examined; 5 species of Syphacia (Nematoda: Oxyuridae), Aspiculuris tetraptera (Nematoda: Heteroxynematidae), and Rodentolepis nana (Cestoidea: Hymenolepididae) were collected. Concurrent infection with 3 pinworm species, Syphacia mesocriceti , Syphacia stroma , and Syphacia peromysci , was recorded for the first time in the golden hamster, Mesocricetus auratus . Syphacia mesocriceti was also identified in the desert hamster, Phodopus roborovskii , and S. peromysci was recovered from the fat-tailed gerbil, Pachyuromys duprasi , and the Cairo spiny mouse, Acomys cahirinus . From the pygmy mouse, Mus minutoides , an undetermined species closely resembling Syphacia megaloon and Syphacia ohtaorum , both parasitic in Mus spp., was collected. Females of another undetermined Syphacia sp. were observed in the greater Egyptian gerbil, Gerbillus pyramidum . All of the host– Syphacia associations, except S. mesocriceti in the golden hamsters, were recorded for the first time. It is suggested that overlapping breeding situations provided the opportunity for host switching by the pinworms.

Strongyloides callosciureus n. sp. (Nematoda: Rhabditoidea), from Asian sciurids, is described based on morphology, morphometry, and the small and large subunit (SSU/LSU) ribosomal RNA gene (rDNA) sequences. This new species was collected from Pallas's squirrels ( Callosciurus erythraeus ) in the central part of mainland Japan (Honshu), which were originally introduced from Taiwan some decades ago, and plantain squirrels ( Callosciurus notatus ) imported from Malaysia as personal pets. For comparison, Strongyloides robustus Chandler, 1942 was collected from American red squirrels ( Tamiasciurus hudsonicus ) and southern flying squirrels ( Glaucomys volans ) imported from the United States as personal pets. The parasitic females found in North American and Asian sciurids shared some key morphological features such as the ovary running spirally around the gut, and the shapes of the stoma in the apical view and the tail. However, morphometric features of parasitic females in North American and Asian sciurids differed significantly from each other; the former was larger than the latter, and the relative position of the vulva to the whole body length from the mouth was different. The SSU/LSU rDNA sequences supported the division of sciurid Strongyloides isolates by geographical distribution of the host and morphological features, leading us to propose the erection of new species.

Strongyloides procyonis Little, 1966 was detected about 45 years ago in raccoons ( Procyon lotor ) of southern Louisiana, U.S.A., and was demonstrated experimentally to cause creeping eruption and a short-lived intestinal infection in a healthy human volunteer. After its description and demonstration of its pathogenicity in humans, S. procyonis has not been found in raccoons in North America despite repeated surveys. During a survey on feral raccoons in Japan, S. procyonis parasitic females were identified in 66 (28.3%) of 233 raccoons collected between May 2004 and January 2005. The number of parasitic females recovered from individual raccoons was 1–197 (geomean, 3.2). Both the morphological features and the nucleotide sequences of the small and large subunit ribosomal RNA genes (SSU/LSU rDNA) of S. procyonis closely resembled those of zoonotic Strongyloides stercoralis . The sequences of internal transcribed spacer (ITS)1 and 28S rDNA could differentiate clearly these 2 species. Awareness of S. procyonis in raccoons in North America and other places worldwide where raccoons are introduced and naturalized is important to assess the epidemiological significance of this potentially zoonotic helminth species.

A colony of Japanese macaques ( Macaca fuscata fuscata ) kept by a safari-style zoo in Japan experienced 9 sporadic cases of fatal neurological diseases, such as epilepsy and posterior paralysis, during the 12 yr from 1989 to 2001. This macaque colony consisted of approximately 30 animals, on average, during this period, and the macaques shared their living space with 11 American black bears ( Ursus americanus ) harboring zoonotic roundworms ( Baylisascaris transfuga ). Close to this enclosure, a cote for 2–3 raccoons ( Procyon lotor ) was placed, and raw sewage from this cote ran into a shallow drain in the area for macaques and bears. However, fecal examinations in recent years did not detect the infection of raccoons with zoonotic roundworms ( Baylisascaris procyonis ). Postmortem histological examination of the latest 2 ill macaques detected multifocal malacia in the brain; 2 ascarid larvae of 60 μm maximum width were encapsulated in the cerebrum and lungs of 1 of the animals. To determine the causative ascarid species of the fatal larva migrans, we analyzed 2 additional encapsulated Baylisascaris larvae collected from formalin-fixed lungs by morphological and molecular approaches. This sporadic outbreak is the second record of Baylisascaris larva migrans in animals in Japan.

Raccoon roundworms ( Baylisascaris procyonis ) and other Baylisascaris species cause patent or latent larva migrans (LM) in a variety of mammals and birds, including humans. It is not clear whether LM by Baylisascaris transfuga , roundworms of bears, is associated with clinical neurological disorders. To clarify this issue, ICR and BALB/c mice as well as Mongolian jirds ( Meriones unguiculatus ) were orally inoculated with 2,000–5,000 embryonated eggs of B. transfuga . In mice, the ascarid caused symptomatic LM of limited extent and duration, whereas the infection was fatal in jirds; i.e., they exhibited general signs such as severe depression and emaciation on days 8–11 postinfection (PI) and died, or they developed progressive and fatal neurological disorders after day 14 PI. Histological examination showed B. transfuga larvae in the brain of all mice and jirds examined, and the larvae collected from them developed to a size comparable with that of B. procyonis . There existed, however, critical differences in host reactions against larvae localized in the brain of mice and jirds; B. transfuga larvae found in mice were surrounded by granulomatous reactions and immobilized, whereas larvae found in jirds were free from any host reaction and mobile, causing extensive malacia.

Twin, white-fronted marmosets ( Callithrix geoffroyi ) born and raised in a zoo in Japan died at 7 mo of age. Several encapsulated nematode larvae were detected in the intestinal wall, as well as a few in the mesenteric lymph nodes of 1 of the twins. In the other marmoset, no encapsulated nematode larva was detected in the organs, but many adult Pterygodermatites nycticebi were found in the intestinal lumen. In the past 5 yr, 5 primates kept in the same zoo, i.e., 1 squirrel monkey ( Saimiri sciureus ), 2 Pygmy marmosets ( Cebuella pygmaea ), 1 Senegal galago ( Galago senegalensis ), and 1 cotton-top tamarin ( Saguinus oedipus ), died from heavy infestation with the same nematode. A few migrating larvae of the rictulariid were also identified histologically in the intestinal wall and liver of the cotton-top tamarin. Although no other primate currently held in the same zoo was infected with the rictulariid, German cockroaches ( Blattella germanica ) collected with traps near marmoset cages had encapsulated P. nycticebi larvae, indicating latent perpetuation of the life cycle of this rictulariid species in the zoo premises. Our results indicated that encapsulation or migration of third-stage larvae of P. nycticebi might occur accidentally in the organs of callithrichid primates.

Intermediate filaments (IFs) make up the cytoskeleton of most eukaryotic cells. In vertebrates, a number of IF proteins have been identified, showing distributions unique to tissue or cell type. Information on helminth IFs is limited to some nematode species. To observe immunofluorescent localization of IFs in helminth tissues, we selected a murine hybridoma clone producing IgM antibody to multiple types of mammalian IF proteins and examined cross-reactivity to helminth proteins. The selected monoclonal antibody (HUSM-9) cross-reacted well with IFs from nematode species such as Toxocara canis, Dirofilaria immitis, Anisakis simplex, and Trichinella britovi ; strong immunofluorescence on cryostat sections was detected in the hypodermis, cords, body muscle, smooth muscle of the uterus, and other epithelial structures. In platyhelminths, i.e., adult Schistosoma mansoni, larval Taenia taeniaeformis, adult Taenia crassiceps, and Echinococcus multilocularis protoscolex, the reactivity was weaker than in nematodes, and localized in the body wall muscle and subtegumental tissue. Western blotting of 8 M urea extracts of parasites with the antibody detected a pair of clear bands in nematodes but not in S. mansoni or the cestodes. These results might be explained by sparse distribution of IFs in platyhelminths, or low affinity of the used antibody to platyhelminth IF proteins, or both.