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EN
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.
EN
The aim of this article is to give an overview of the molecular methods used for detecting helminth parasites in the environment, their advantages as well as their limitations with examples of successful applications of these methods in different helminth classes. Molecular techniques represent a wide range of sensitive diagnostic tools in parasitology. Environmental monitoring is an essential element of epidemiological studies. Many helminth parasites depend on the environment for the completion of their life cycle. The abundance of their transmissive stages determines infection risks. The application of highly specific and sensitive monitoring methods allows gathering more precise epidemiological data.
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