It is uncertain when the head collar and collar spines of Isthmiophora hortensis (Digenea: Echinostomatidae), a zoonotic echinostome species in Far Eastern Asia, develop during its larval stages. In this study, the appearance of the head collar and collar spines was studied using light and scanning electron microscopy in cercariae and metacercariae experimentally obtained from freshwater snails (Lymnaea pervia) and tadpoles (Rana nigromaculata), respectively. The cercariae were shed from the snail on day 30 after exposure to laboratory-hatched miracidia. Metacercariae were obtained from the experimental tadpoles at 3, 6, 12, 15, 20, 24, 26, and 30 h after exposure to the cercariae. The head collar was already visible in the cercarial stage, although its degree of development was weak. However, collar spines did not appear in the cercarial stage and even in the early metacercarial stage less than 24 h postinfection in tadpoles. Collar spines became visible in the metacercariae when they grew older than 24 h. It was concluded that the head collar of I. hortensis developed early in the cercarial stage, but the development of collar spines did not occur until the worms became 24-h-old metacercariae in our experimental setting. Counting the number of collar spines was concluded as an unfeasible diagnostic method for I. hortensis cercariae when they are shed from the snail host.
The objective of this study was to evaluate the efficacy of an imidacloprid 10% and flumethrin 4.5% polymer matrix collar against the developmental stages of Haemaphysalis longicornis infesting dogs using the hair from treated dogs in a semi-in-vitro assay set. When incubated with 0.5 g of the hair collected from the dogs installed with the drug-embedded collar after 10 days, average death rate of the larval, nymphal, and adult H. longicornis was 21.5%, 77.9%, and 100% at 30 min, 1 hr, and 2 hr, respectively. This study showed the larval stages as well as the nymphal and adult stages of H. longicornis ticks are killed upon contact with the hair from dogs treated with the collar within 2 hr.
Echinostoma mekongi n. sp. (Digenea: Echinostomatidae) is described based on adult flukes collected from humans residing along the Mekong River in Cambodia. Total 256 flukes were collected from the diarrheic stool of 6 echinostome egg positive villagers in Kratie and Takeo Province after praziquantel treatment and purging. Adults of the new species were 9.0-13.1 (av. 11.3) mm in length and 1.3-2.5 (1.9) mm in maximum width and characterized by having a head collar armed with 37 collar spines (dorsal spines arranged in 2 alternative rows), including 5 end group spines. The eggs in feces and worm uterus were 98-132 (117) μm long and 62-90 (75) μm wide. These morphological features closely resembled those of Echinostoma revolutum, E. miyagawai, and several other 37-collar-spined Echinostoma species. However, sequencing of the nuclear ITS (ITS1-5.8S rRNA-ITS2) and 2 mitochondrial genes, cox1 and nad1, revealed unique features distinct from E. revolutum and also from other 37-collar-spined Echinostoma group available in GenBank (E. bolschewense, E. caproni, E. cinetorchis, E. deserticum, E. miyagawai, E. nasincovae, E. novaezealandense, E. paraensei, E. paraulum, E. robustum, E. trivolvis, and Echinostoma sp. IG). Thus, we assigned our flukes as a new species, E. mekongi. The new species revealed marked variation in the morphology of testes (globular or lobulated), and smaller head collar, collar spines, oral and ventral suckers, and cirrus sac compared to E. revolutum and E. miyagawai. Epidemiological studies regarding the geographical distribution and its life history, including the source of human infections, remain to be performed.
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Echinostoma flukes armed with 37 collar spines on their head collar are called as 37-collar-spined Echinostoma spp. (group) or ‘Echinostoma revolutum group’. At least 56 nominal species have been described in this group. However, many of them were morphologically close to and difficult to distinguish from the other, thus synonymized with the others. However, some of the synonymies were disagreed by other researchers, and taxonomic debates have been continued. Fortunately, recent development of molecular techniques, in particular, sequencing of the mitochondrial (nad1 and cox1) and nuclear genes (ITS region; ITS1-5.8S-ITS2), has enabled us to obtain highly useful data on phylogenetic relationships of these 37-collar-spined Echinostoma spp. Thus, 16 different species are currently acknowledged to be valid worldwide, which include E. revolutum, E. bolschewense, E. caproni, E. cinetorchis, E. deserticum, E. lindoense, E. luisreyi, E. mekongi, E. miyagawai, E. nasincovae, E. novaezealandense, E. paraensei, E. paraulum, E. robustum, E. trivolvis, and Echinostoma sp. IG of Georgieva et al., 2013. The validity of the other 10 species is retained until further evaluation, including molecular analyses; E. acuticauda, E. barbosai, E. chloephagae, E. echinatum, E. jurini, E. nudicaudatum, E. parvocirrus, E. pinnicaudatum, E. ralli, and E. rodriguesi. In this review, the history of discovery and taxonomic debates on these 26 valid or validity-retained species are briefly reviewed.
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