Bithyniid snails as hosts of Opisthorchiidae and Notocotylidae in the south of Western Siberia, Russia.

Opisthorchiasis and related infections are persistent and substantial public health burdens from eastern Europe to southeastern and northern Asia. Snails of the family Bithyniidae act as first intermediate hosts not only for the trematodes of the family Opisthorchiidae but also for those of the family Notocotylidae. There are certain morphological similarities between the aforementioned trematode cercariae. In order to find natural local foci of opisthorchiasis, which are established primarily according to the presence of infected bithyniid snails at the area under examination, it is crucial to correctly identify the rediae and cercariae of the trematodes. The aim of our investigation was to evaluate the role of bithyniid snails in the transmission of Opisthorchiidae and Notocotylidae in ecosystems in the south of Western Siberia. We have been studying the process of bithyniid snail dissemination in Western Siberia and examining their infection by trematodes from 1994 until now. A total of 16,213 bithyniid snails in 23 water bodies (in 28 localities) of four major basins situated in the Novosibirsk region were inspected for trematode infestation. Long-term research has been conducted in the Kargat river estuary and the Ob river floodplain for 15 and 25 years, respectively. In both water bodies, the prevalence of notocotylid and opisthorchiid trematodes was positively correlated with the global land-ocean temperature index. Trematode parthenitae were identified if there were mature cercariae capable of leaving the shell of the host snail on their own. Identification to the species of opisthorchiid cercariae was verified using molecular genetic analysis methods targeting ITS2 locus. Opisthorchis felineus and Metorchis bilis, two opisthorchiid species that are potentially perilous to human health, were found in bithyniids in the Novosibirsk region. The mean prevalence of infection with notocotylid trematodes in bithyniid snails was higher than the corresponding prevalence of infection with opisthorchiid trematodes. The results of this study can be used to identify and predict natural foci of epidemiologically and/or epizootically dangerous diseases.

Molecular Identification of the Human Pathogen Amphimerus sp. in the Freshwater Snail Aroapyrgus sp. in Ecuador.

Here, we report for the first time the snail intermediate host for the Amphimerus liver fluke, a foodborne trematodiasis. In Ecuador, Amphimerus of the Opisthorchiidae family, infects humans, cats, and dogs, in the tropical Pacific-coast region. Opisthorchiidae comprising also Clonorchis sinensis, Opisthorchis sp., and Metorchis sp., have complex life cycles involving a definitive and two intermediate hosts. We identified morphologically and investigated the presence and prevalence of Amphimerus cercaria and DNA in freshwater snails collected in a human-amphimeriasis endemic region in Ecuador, extracted DNA from snail tissue and emerged cercariae, performed real-time polymerase chain reaction (PCR) with the newly developed primers and probe amplifying the Amphimerus ribosomal internal transcribed spacer 2 (ITS2) region, and sequenced the amplified DNA fragment. We collected 2,800 snails, characterized four species Aroapyrgus sp., Melanoides tuberculata, Biomphalaria cousini, and Aplexa marmorata, isolated three cercariae morphotypes. Of the 640 snails analyzed by qPCR, only Aroapyrgus and one of the three cercariae resulted positive, at a 15% infection prevalence. Polymerase chain reaction revealed that the Aroapyrgus snail and cercaria-morphotype-3 corresponded to Amphimerus, but not to C. sinensis, Fasciola hepatica, or Paragonimus mexicanus. The sequence of amplified DNA product matched that of human-isolated Amphimerus. This finding constitutes the first documentation that Aroapyrgus sp. is the first intermediate host for the Amphimerus sp. that infect humans in Ecuador. The ITS2-gene PCR and sequencing analysis demonstrated a high prevalence of snail infection and proved useful for detecting the infection in snails, which findings can help the establishment of suitable control programs against transmission in any endemic region of interest.

Description of a new species, Cryptocotyle lata sp. nov., and discussion of the phylogenetic relationships in Opisthorchioidea.

Adult Cryptocotyle lata sp. nov. worms were obtained from experimental studies. In the Russian southern Far East, the life cycle of this parasite is carried out using freshwater snails (Boreoelona ussuriensis), freshwater fish, and birds as the first intermediate, second intermediate, and definitive hosts, respectively. The morphological indices of C. lata sp. nov. are closest to Cryptocotyle concava; however, these two species differ in terms of their sizes of body, oral and ventral suckers, eggs, and the shape of their testes and ovaries. Analysis of the life cycles of the Cryptocotyle representatives suggested that C. concava were at least two cryptic species, one of which circulates using brackish water Hydrobia snails, and the other using freshwater Amnicola snails as the first intermediate hosts. Molecular data (i.e., the 28S gene and the second internal transcribed spacer (ITS2) of rDNA) were used to analyze the phylogenetic relationships of C. lata sp. nov. and other representatives of Opisthorchioidea. The long repeats and secondary structure of the ITS1 region were studied. Representatives of the Opisthorchiidae and several species from Heterophyidae (including the genus Cryptocotyle) were found to have molecular features that suggested that these species belonged to Opisthorchiidae. At the same time, the genetic relatedness of worms, which are united in common clusters on phylogenetic trees, is consistent with the use of the first intermediate hosts from different taxonomic groups in their life cycles; namely, snails of the Truncatelloidea are hosts of trematodes from a cluster with Opisthorchiidae and a number species of the family Heterophyidae, while snails of the Cerithioidea are hosts of worms from a cluster that includes only the Heterophyidae. In addition, the results of genetic studies indicate that Clonorchis sinensis, Metorchis ussuriensis, Metorchis bilis, Metorchis xanthosomus, and Metorchis orientalis should be included in the genus Opisthorchis.

[Mucoid glands of Cercaria baushii 6 (Xiphidiocercariae: Cercariae microcotylae)].

Mucoid glands of Cercaria baushii 6 (Xiphidiocercariae: Cercariae microcotylae Luhe, 1909) were examined. Gland cells differ from each other in the time of differentiation and in size. A brief sketch on the morphology of parthenites is also given.

Opisthorchis felineus and Metorchis bilis are the main agents of liver fluke infection of humans in Russia.

Liver fluke infections are gradually transforming from a local problem of individual geographic regions to a widespread problem. The observed expansion is likely to be connected with the ever-increasing intensity of traffic flow and migration of the infected carriers between cities, regions, and countries. Opisthorchis felineus, the trematode belonging to the family Opisthorchiidae, is a well known causative agent of the infection called opisthorchiasis. Metorchis bilis, also a member of the family Opisthorchiidae, causes metorchiasis, a disease very close to opisthorchiasis in symptomatology. Genetic markers can be used to develop methods for differential diagnostics of these diseases. However, the questions connected with epidemiology of these trematode infections, their clinical characteristics, prognosis and therapy remain open. This review briefs the general biological characteristics of O. felineus and M. bilis persisting in various countries of Eurasia, their geographical range, epidemiology and molecular diagnostics of these liver flukes.

Molecular and morphometric study of metacercariae and adults of Pseudamphistomum truncatum (Opisthorchiidae) from roach (Rutilus rutilus) and wild American mink (Mustela vison).

The presence of metacercariae and adults of the trematode Pseudamphistomum truncatum in roach and mink, respectively, was recorded in Lake Fure North of Copenhagen, Denmark. This zoonotic digenean opisthorchiid represents a threat to humans due to its ability to infect the biliary system following ingestion of inadequately processed infected fish. Therefore precise species identification of infective metacercariae in fish used for human consumption is essential. Due to the relatively limited information on metacercarial identity obtained by morphometric studies a series of molecular techniques were used to link the larval parasite in fish with the un-equivocally diagnosed adults in the biliary system of the mink. By the use of carefully selected polymerase chain reaction (PCR) primers and subsequent sequencing of the ITS region from both metacercariae and adults full sequence identity of both metacercariae and adults were confirmed. The presence of this parasitosis in fish from a lake used for both commercial and recreational fisheries call for hygienic alerts in order to prevent accidental human infection with this opisthorchiid.

Amphimerus bragai n. sp. (Digenea: Opisthorchiidae), a parasite of the rodent Nectomys squamipes (Cricetidae) from Minas Gerais, Brazil.

Amphimerus bragai n.sp. (Digenea, Opisthorchiidae) from the bile ducts of a rodent from the State of Minas Gerais, Brazil, Nectomys squamipes (Cricetidae), is described. The new species as studied by both light and scanning electron microscopy. A table is presented comparing the measurements of the new species with those of A. lancea (Diesing, 1850) and A. vallecaucensis Thatcher; 1970, parasites of dolphins and marsupials, respectively. The new species is similar in size and body form to A. vallecaucensis from which it differs in having a vitellarium that extends to the acetabulum while that of the former species are limited to the posterior one-third of the body. Additionally, the new species is from a rodent.

Amphimerus bragai n.sp. (Digenea: Opisthorchiidae), a parasite of the rodent Nectomys squamipes (Cricetidae) from Minas Gerais, Brazil

Amphimerus bragai n.sp. (Digenea, Opisthorchiidae) from the bile ducts of a rodent from the State of Minas Gerais, Brazil, Nectomys squamipes (Cricetidae), is described. The new species was studied by both light and scanning electron microscopy. A table is presented comparing the measurements of the new species with those of A. lancea (Diesing, 1850) and A. vallecaucensis Thatcher, 1970, parasites of dolphins and marsupials, respectively. The new species is similar in size and body form to a A. vallecaucensis from which it differs in having a vitellarium that extends to the acetabulum while that of the former species are limited to the posterior one-third of the body. Additionally, the new species is from a rodent.

[Growth and development of Metorchis orientalis in chicks and its adult morphology].

In order to observe the infectivity, growth and development and adult morphology of Metorchis orientalis, a total of 40 chicks were experimentally infected with 100 metacercariae respectively, collected from Pseudorasbora parva. The worms of various developmental stages were recovered from chicks at 1.5, 3, 5, 7, 9, 11, 14 and 21 days after infection, and they were prepared for morphological observations and measurements. All of the worms were found in the gallbladders of chicks, and their recovery rate was 32% in average. The growth of the body was rapid from 9 to 11 days after infection. The genital primordia appeared in 1.5 and 3-day old worms, and ovary and testes were first observed in 5-day old worms. Thereafter, genital organs gradually matured and completed up to 11 days after infection. The adult worm was leaf-like, and possessed a convoluted tubular seminal vesicle, an ovoid ovary, a sac-like seminal receptacle, 2 lobed-testes and follicular vitellaria. Eggs were 31.9 x 15.3 microns in average size, ellipsoid to elliptical in shape and possessed abopercular thickenings. From the above results, it is concluded that M. orientalis grows in sigmoid pattern in chicks, and their genital organs fully mature between days 9 and 11. It is also confirmed that a chick is a new definitive host of M. orientalis.

[Sensillae of cercariae of Haplorchis taichui (Digenea, Heterophyidae). Discussion of the systematics of the Opisthorchioidea]. / Les sensilles de la cercaire d'Haplorchis taichui (Digene, Heterophyidae). Discussion sur la systématique des Opisthorchioidea.

Cercarial chaetotaxy of Haplorchis taichui (Haplorchiinae, Heterophyidae) is described and compared to that of ten other Opisthorchioidea: three belonging to the Acanthostomidae (Acanthostominae), one to the Opisthorchiidae (Opisthorchiinae) and six to the Heterophyidae (among these, two Centrocestinae, two Apophallinae, one Euryhelminthinae and one Haplorchiinae). Cercarial chaetotaxy allow us to distinguish the considered species and genera, but does not support division of the Opisthorchioidea into families as these have been defined based on morphology of adults; it suggests that the Heterophyidae is heterogenous; it allows us to recognize the Opisthorchioidea characterized by a loss of hexaradial symmetry on the first cephalic cycle of sensillae and by the presence of two to five pairs of sensillae on a tail with proximal excretory apertures.