Susceptibility of Bithynia inflata mollusks from discrete populations for Opisthorchis felineus infestation from different foci of opisthorchiasis

Experiments were conducted on crossed infection of the specific intermediate host of Opisthorchis, molluscs of Bithynia inflata from different and remote populations, with eggs of O. felineus from different nidi of the disease. The strains of the agent were found to be heterogeneous that is expressed in different degree of compatibility with the intermediate host.     

Heterogeneity of populations of the mollusc Bithynia inflata according to the degree of trematode infestation in opisthorchiasis foci

Results of studies of differences in the infection rate of natural populations of Bithynia inflata with trematodes (including Opisthorchis felineus) in the nidi of opisthorchiasis in the Sumsk and Chernigov Districts of the Ukrainian SSR and Tomsk District of the RSFSR are given. It has been shown that in the Ukrain over 1/3 of all populations of the specific intermediate host of opisthorchis does not take part in the circulation of the agent due to fundamentally different reasons: resistence to the infection and hyperinfection with parthenites of trematodes.     

Comparative analysis of karyotypes of Opisthorchis felineus from West Siberia

Karyotype of the trematode Opisthorchis felineus from two populations of the Ob and the Tom’ rivers basins in the southeast of the West Siberian plain was investigated. Comparative analysis of the relative length and centromere indices of seven chromosomes showed that they are identical to literature data on the chromosome sets of this trematode species from the lower and upper courses of the Irtysh River. The quality of chromosome spreads improved the former knowledge of the karyotype structure of this species, in particular, the 6th and 7th chromosomes.     

The ecology of the Bithynia first intermediate hosts of Opisthorchis viverrini

Opisthorchiasis, together with its associated cholangiocarcinoma, is one of the most important human parasitic diseases on continental Southeast Asia. A great deal of epidemiological data from humans is available on this disease, particularly from the northeast of Thailand, however, only limited information is available on those aspects of the life cycle relating to its Bithynia (Gastropoda) and cyprinid fish intermediate hosts. Here we review the information which is available on the Bithynia hosts of Opisthorchis viverrini. Only one major ecological study has been carried out at one site on a single species of Bithynia. We show not only that detailed ecological studies are required to clarify the epidemiology of opisthorchiasis, but also that the taxonomic status of the Bithynia species transmitting O. viverrini requires clarification.     

Susceptibility of different populations of ticks to entomopathogenic fungi

This study aimed to evaluate the in vitro effect of the entomopathogenic fungi Metarhizium anisopliae sensu lato (s.l.) and Beauveria bassiana sensu lato (s.l.) on two distinct populations of Rhipicephalus microplus, from two different experimental farms. Bioassays were performed with engorged females, eggs and larvae. Fungal infection was evaluated based on biological parameters of treated engorged females, percentage of hatch from treated eggs, and percentage of mortality and mean lethal time (LT90) of treated larvae. When the treatments were compared between the two populations, there were significant differences in the following parameters: pre-oviposition period, hatching period and egg production index. Moreover, the results showed that B. bassiana s.l., isolate Bb 986, was more virulent than M. anisopliae s.l., isolate 959, for engorged females, showing a control percentage of 49%. In the bioassay with eggs, the hatching percentage ranged from 3.1% to 49.5% in one population and from 3.4% to 42.7% in the other, with no significant difference between the two populations. In the bioassay with unfed larvae, the mortality percentage ranged from 91.8% to 98.7% in one population and from 71.0% to 94.0% in the other. The LT(90) varied from 19.52 to 27.51 days in one of the populations and 22.89 to 37.31 days in the other. These results suggest that populations of R. microplus show distinct variation in their susceptibility to B. bassiana s.l. and M. anisopliae s.l.     

Effect of light intensity on Opisthorchis viverrini cercarial shedding levels from Bithynia snails--a preliminary study

Opisthorchis viverrini requires Bithynia snails as the first intermediate host and cyprinid fish as the second intermediate host. Very low natural infection rates have been reported in Bithynia snails, but very high rates have been found in cyprinid fish in the same endemic region. This study investigated the effect of light intensity, the most important stimulus, on the quantity of O. viverrini cercariae shed from naturally infected Bithynia (Digoniostoma) siamensis goniomphalos snails. Snails were evaluated for cercariae output every hour after exposure to various light intensities for a total period of 7 h. The same infected snail was tested under different intensities of light: in the dark, and at 1000, 3000 and 5000 lx. The data showed that under exposure to 1000 and 3000 lx of light, the average percentage and number of cercariae released were higher than that exposed to 5000 lx during the first 2 h of the experiment. In contrast, under higher illumination (5000 lx) a longer time (6 h) was required to stimulate the peak emergence of cercariae. Darkness was not able to induce O. viverrini cercariae emergence. Among the three intensities of light, exposure at 1000 lx induced the highest average number of released cercariae per snail and the highest percentage of cercarial emergence within the first 2 h (125, 54.86%), followed by exposure at 3000 lx (69, 25.58%) and 5000 lx (12, 7.78%). The results suggest that the light intensity of 1000 lx for 2 h would be optimal for O. viverrini cercarial shedding from naturally infected B. (D.) siamensis goniomphalos snails.     

Experimental grounds of a new concept of Opisthorchis felineus infection formation in a definitive host

In the course of experiments in was found out that golden hamsters having the bilious duct operationally blocked display receptivity to the infection with the non-excysted Opisthorchis felineus metacercariae passed to the stomach. Excysted metacercariae injected to the system of the portal vein settle down in the bilious pathways of the liver and develop there up to the adult stage. In vitro, the metacercariae survive in the blood serum of the intact golden hamsters during one day. Based on the experiments, it is hypothesized that the early stage of O. felineus infection in the bilious duct of definitive hosts is performed by means of hematogenic migration of metacercariae through the portal veins system from the mucous layer of the alimentary tract of the host.     

Comparative histopathology of Opisthorchis felineus and Opisthorchis viverrini in a hamster model: an implication of high pathogenicity of the European liver fluke

European liver fluke (Opisthorchis felineus) and Asian liver fluke (Opisthorchis viverrini) are similar in morphology but comparative pathology of the infections has not been described. We therefore did comparative histopathology of both parasites in an experimental animal model. The study was conducted in 3 groups of 105 Syrian golden hamsters; the first and second groups fed with 50 metacercariae of O. felineus (OF) or O. viverrini (OV) and the last group was uninfected controls. Five hamsters in each group were euthanized on weeks 1, 2, 4, 8, 12 and 24 post-infection. The liver tissue was fixed and processed for routine histopathology and immunohistochemistry for proliferation markers (BrdU or PCNA). Overall, the liver histopathology of O. felineus and O. viverrini infection was generally similar. However, various histopathogical features including intense inflammation, fibrosis, biliary and goblet cell hyperplasia and dysplasia occurred earlier in the OF group. In addition, the existence of precancerous lesions such as cholangiofibrosis in a long-term infection was observed only in this group. O. felineus is larger in size than O. viverrini which, together with its excreted and secreted antigens, likely is crucial in the induction of liver fluke induced disease. The differences in nature and timing of the histopathological profile indicate that opisthorchiasis caused by the European liver fluke O. felineus is more pathogenic than its Asian relative O. viverrini.     

The complete mitochondrial genomes of the liver flukes Opisthorchis felineus and Clonorchis sinensis (Trematoda)

The complete mitochondrial genomes of the parasitic trematodes Opisthorchis felineus and Clonorchis sinensis (family Opisthorchiidae) were fully sequenced in order to develop markers for DNA diagnostics of the liver flukes infection, molecular ecology, population and phylogenetic studies. The complete sequences of mitochondrial genomes of these species comprise 14,277 and 13,875 bp, respectively, and are thus the shortest trematode mitochondrial genomes sequenced to date. The gene content and arrangement are identical to that of Fasciola hepatica. ATG and GTG are used as the start-codons and TAG and TAA are used as the stop-codons. The stop-codon TAG of the C. sinensis nad1 gene overlap by 1 nt with the downstream tRNA-Asn gene. Alternative structures for the Ser(UCN) tRNAs were found for both species. The noncoding control regions are separated into two parts by the tRNA-Gly gene and contain neither tandem repeats, which are characteristic for trematode control regions, nor secondary structures. In conclusion, the complete mitochondrial DNA sequences of O. felineus and C. sinensis will serve as a resource for comparative mitochondrial genomics and systematic studies of parasitic trematodes.     

Effect of an Opisthorchis infestation on implantation growth of the liver epithelium

The features of proliferation and differentiation of the liver ductular epithelium from animals infected with opisthorchis were specified in experimental implantation growth. They manifested themselves in disorders of the mitotic regimen and intercellular interactions, in atypical proliferations and primary glandular metamorphism of regenerating cells.     

Identification of novel natural inhibitors of Opisthorchis felineus cytochrome P450 using structure-based screening and molecular dynamic simulation

Opisthorchis felineus is the etiological agent of opisthorchiasis in humans. O. felineus cytochrome P450 (OfCYP450) is an important enzyme in the parasite xenobiotic metabolism. To identify the potential anti-opisthorchid compound, we conducted a structure-based virtual screening of natural compounds from the ZINC database (n = 1,65,869) against the OfCYP450. The ligands were screened against OfCYP450 in four sequential docking modes that resulted in 361 ligands having better docking score. These compounds were evaluated for Lipinski and ADMET prediction, and 10 compounds were found to fit well with re-docking studies. After refinement by docking and drug-likeness analyses, four potential inhibitors (ZINC2358298, ZINC8790946, ZINC70707116, and ZINC85878789) were identified. These ligands with reference compounds (itraconazole and fluconazole) were further subjected to molecular dynamics simulation (MDS) and binding energy analyses to compare the dynamic structure of protein after ligand binding and the stability of the OfCYP450 and bound complexes. The binding energy analyses were also calculated. The results suggested that the compounds had a negative binding energy with ?259.41, ?110.09, ?188.25, ?163.30, ?202.10, and ?158.79 kJ mol^?1 for itraconazole, fluconazole, and compounds with IDs ZINC2358298, ZINC8790946, ZINC70707116, and ZINC85878789, respectively. These lead compounds displayed significant pharmacological and structural properties to be drug candidates. On the basis of MDS results and binding energy analyses, we concluded that ZINC8790946, ZINC70707116, and ZINC85878789 have excellent potential to inhibit OfCYP450.     

Susceptibility and physiological responses of Jatropha curcas accessions to broad mite infestation

The broad mite Polyphagotarsonemus latus is a key pest of physic nut (Jatropha curcas L.). The purpose of this study was to identify physic nut accessions that are less susceptible to P. latus, in support of the breeding program of J. curcas. We first evaluated population growth rate and injury symptoms of P. latus on different J. curcas accessions and then carried out physiological analyses on P. latus-infested and uninfested accessions. From the germplasm bank of the Federal University of Viçosa, 15 physic nut accessions with high seed oil content, with different genetic background, were tested. The following traits were evaluated: instantaneous population growth rate of P. latus (r _i ), injury symptoms, relative leaf water content, specific leaf area, gas exchange, photosynthetic pigments, nitrogen and biomass of the aerial part. Significant differences were observed for P. latus population growth rate and injury symptoms among accessions. A positive correlation between P. latus growth rate and injury was found. The UFVJC72 accession stood out as the more resistant, considering P. latus growth rate and injury symptoms, compared with most accessions. Physiological responses did not vary among accessions, but did between infested and uninfested plants. In P. latus-infested plants, net photosynthesis was on average 50.5 % lower than in uninfested plants, whereas stomatal conductance and transpiration decreased by 46.2 and 51.6 %, respectively.     

Low Genetic Diversity in Wide-Spread Eurasian Liver Fluke Opisthorchis felineus Suggests Special Demographic History of This Trematode Species

Opisthorchis felineus or Siberian liver fluke is a trematode parasite (Opisthorchiidae) that infects the hepato-biliary system of humans and other mammals. Despite its public health significance, this wide-spread Eurasian species is one of the most poorly studied human liver flukes and nothing is known about its population genetic structure and demographic history. In this paper, we attempt to fill this gap for the first time and to explore the genetic diversity in O. felineus populations from Eastern Europe (Ukraine, European part of Russia), Northern Asia (Siberia) and Central Asia (Northern Kazakhstan). Analysis of marker DNA fragments from O. felineus mitochondrial cytochrome c oxidase subunit 1 and 3 (cox1, cox3) and nuclear rDNA internal transcribed spacer 1 (ITS1) sequences revealed that genetic diversity is very low across the large geographic range of this species. Microevolutionary processes in populations of trematodes may well be influenced by their peculiar biology. Nevertheless, we suggest that lack of population genetics structure observed in O. felineus can be primarily explained by the Pleistocene glacial events and subsequent sudden population growth from a very limited group of founders. Rapid range expansion of O. felineus through Asian and European territories after severe bottleneck points to a high dispersal potential of this trematode species.     

Susceptibility of cell populations to transduction by retroviral vectors

Retroviral transduction efficiency is related to the multiplicity of infection and the physiological state of the target cells. It is generally not known what proportion of a cell population is susceptible to transduction. We used coinfection with two retroviral vectors containing the marker genes for green fluorescent protein and the truncated human nerve growth factor receptor. In the CD34+ cell line TF-1 or human primary CD34+ hematopoietic progenitor cells, it was found that cells transduced with one vector had a better than random chance of transduction by the other vector. A probability model was developed to estimate target cell susceptibility; susceptibility was calculated as the product of the proportions of transgene-positive cells divided by the proportion of double-positive cells. By using this relationship, it was found that susceptibility was related to the target cell type and culture conditions but not the retroviral titer or the retroviral packaging envelope protein used in this study. Cotransduction with two vectors is a relatively simple procedure that provides a means to assess the maximum transduction level possible in a given cell population.     

Susceptibility of endothelial cells derived from different blood vessels to common viruses

Summary We examined whether endothelial cells derived from different blood vessels vary in their susceptibility to viral infection. Five common viral pathogens of humans (herpes simplex 1, measles, mumps, echo 9, and coxsackie B4 viruses) were evaluated for growth in endothelial cells derived from bovine fetal pulmonary artery thoracic aorta, and vena cava. All five viruses replicated in each type of endothelial cell. There were apparent differences in the quantities of measles and mumps viruses produced in pulmonary artery endothelium compared with thoracic aorta and vena cava when endothelial cells were obtained from different animals. However when pulmonary artery endothelial cells were compared with vena cava cells from the same animal, growth of each virus was similar in the two cell types. Four of the viruses replicated in the various endothelial cells without producing appreciable changes in cell morphology. These results indicate that endothelial cells from different blood vessels are equally susceptible to the human viruses evaluated, and that viral replication can occur without major alteration in cell morphology. Endothelial cells could serve as permissive cells permitting viruses to leave the circulation and initiate infection in adjacent tissues, including subendothelial smooth muscle cells. This work was supported by Public Health Service grants HL28220, HL 29492, and HL 24914 from the National Heart, Lung and Blood Institute, Bethesda, MD.     

The influence of the degree of infestation with trematode parthenites on the structure of penial glands in the mollusks Littorina saxatilis of different age

Histological and confocal microscopy studies of the structure of penial glands in the Littorina saxatilis males were carried out. The examined mollusks belong to two age groups and were at different stages of spontaneous infection with a trematode from the pygmaeus species group (Microphallus piriformes). Based on comparative analysis of microscopic sections of copulative organs in infested and non-infested mollusks, data on the modifications in histological structure ofpenial glands were obtained. From these data we can suggest that the infestation have an influence on all parts of the gland. Decrease of secret production and reduction of muscular capsule (down to its disappearance) were observed. We suppose that changes in the penial glands structure prevent their normal functioning. In the mollusks infested on reaching the sexual maturity, gradual reduction of distal part of reproductive system was observed, while in the mollusks infested before the sexual maturity, development of ancillary part of reproductive system was blocked.     

The role of the association of Opisthorchis felineus maritae with the Epstein-Barr virus in the cytogenetic sequelae of an opisthorchiasis infection

The immunofluorescence analysis of Epstein-Barr virus (EBV) antigens in the body of Opisthorchis felineus (OF) helminths was carried out. It was found that EBV antigens located in eggs of helminth. Adding of OF antigens and/or EBV to the lymphocyte human cultures of healthy donors can induce a cytogenetic damage in cells. A role of EBV location in opisthorchis eggs in chromosome damage and cancer development is discussed.