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Ultrastructure of the digestive gland has been studied in G. truncatula, uninfected and infected with F. hepatica larvae. The digestive gland of G. truncatula consist of seccretory-resorption cells and calcareous cells. On the 20 day post infection the damage to the secretory cells were small and did not inhibit their activity. On the 60 day post infection the gland structure was totally altered: reduced height of secretory cells, lack of Golgi apparatus, myelin-like degradation of mitochondria, reduction and desintegrating of rough endoplasmic reticulum. These damage of secretory and calcareous cell to bring about a total inhibition of their secretory activities.
Fasciolosis, caused by the liver fluke (Fasciola hepatica) is an important issue for both human and animal health. The disease evokes economic losses which are a consequence of impaired animal productivity leading to higher costs of meat and milk production, as well as liver condemnation. The goals of this thesis were to: (1) elaborate a molecular method - PCR for the detection of F. hepatica DNA in intermediate and definite hosts; (2) estimate the usefulness of a recombinated cysteine proteinase produced in E. coli in the form of inclusive bodies in serological diagnosis of F. hepatica infection in definite hosts, using an enzyme-linked immunosorbent assay (ELISA); (3) conduct field research on the prevalence of infection among intermediate and definitive hosts (cattle) in chosen regions of Poland, utilizing the elaborated methods. Based on the results obtained in this study, it was established that it is possible to detect F. hepatica DNA in the feces of definite hosts with the elaborated PCR method. The amplification of a 124 base pair tandem repeat allows the detection of fluke larval stages in intermediate hosts within 12 hours of exposure and F. hepatica infection in definite hosts (by the 5th week in rats, 8th week in sheep and 10th week in cattle). Therefore, the PCR test is more sensitive than traditional microscopic methods. Furthermore, it was determined that, the recombinated cysteine proteinase in the form of inclusive bodies, after solubillization exhibits antigenic properties of the native protein and the ELISA method based on this antigen may be useful as a tool for diagnosing fasciolosis in sheep and cattle, in both serum and milk samples. The test achieves a greater sensitivity and specificity than an ELISA based on native excretory-secretory antigens. The results of field research indicate that Fasciola hepatica is a frequent parasite of cattle in central and eastern Poland. The mean prevalence was 34.86% (±16.95) in all studied areas. The prevalence among intermediate hosts varied greatly (0–100%). The elaborated tests were proved to be valuable, mutually complementing diagnostic tools, applicable to different epidemiological situations.
Hemolymph plasma polypeptides of three pulmonate snail species were analysed by the method of SDS-PAGE followed by Coomassie, silver or Schiff's staining. Polypeptide patterns were also monitored during the course of Fasciola hepatica infection in Galba truncatula and G. palustris. Infection did not result in the induction of specific molecules, but in an augmentation of a few protein fractions, possibly glycoproteins.
In Iran, more than 100 humans and many livestock species have shown to be infected in the northern province of Mazandaran, at the Caspian Sea shore. This picture suggested the need for further multidisciplinary studies to ascertain the extent of the problematics. Three species of lymnaeids were found throughout most of Mazandaran: Lymnaea (Stagnicola) palustris, a secondary intermediate host of Fasciola hepatica; Galba truncatula, the main intermediate host of F. hepatica: and Radix gedrosiana, a member of the auricularia complex transmitting F. gigantica. Environmental microhabitat requirements are different for the three species, but sometimes the three were found in the same locality. The geographical distribution of the three lymnaeids agrees with the distribution of human and animal fascioliasis, differences in fascioliasis prevalences between western and eastern Mazandaran not appearing to be related with lymnaeids. Moreover, the overlapping distributions of G. truncatula and R. gedrosiana also agree with the overlapping distributions of F. hepatica and F. gigantica detected in Mazandaran animals. Although the three lymnaeid species may be found in all the months, interesting differences were detected. Whereas L. (S.) palustris appears to be more or less stable throughout the year, with only one acrophase around June, G. truncatula and R. gedrosiana showed an evident biseasonal distribution, with abundance and population density peaks in spring-beginning of summer and autumn. Such lymnaeid population biseasonality suggests a higher contamination risk by the two fasciolid species for both humans and animals in these two seasons and the corresponding detection of an increase of acute human cases attending the health centres. However, lymnaeid population dynamics do not explain the main peak of human diagnosed patients in the February-March period in Mazandaran. Peculiar culinary habits, as the custom of producing and stocking the traditional "delar" and its use as condiment sauce for many vegetables and food, may explain this chronological disagreement, tacking into account the long viability that fasciolid metacercariae can reach.
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