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Hypoderma sp. - rzadki, nietypowy pasożyt koni

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Liczne gatunki muchówek (Diptera) prowadzą pasożytniczy tryb życia. Spośród nich trzy grupy są najpowszechniejsze i mają największe znaczenie chorobotwórcze u zwierząt hodowlanych i wolno żyjących. Są to muchówki należące do rodziny Oestridae, z podrodzin Gasterophilinae, Hypodermatinae i Oestrinae. W pracy opisano przypadek inwazji muchówek z rodzaju Hypoderma u koni utrzymywanych w klubie jeździeckim w okolicy Krakowa (Polska południowa), znajdującym się w bliskim sąsiedztwie gospodarstwa zajmującego się chowem bydła opasowego. Konie w wymienionym klubie są corocznie odrobaczane jesienią (październik), preparatem zwierającym iwermektynę. U dwóch koni stwierdzono obecność guzów, z otworami zamkniętymi śluzowo-ropną wydzieliną, na kłębie oraz zadzie. Larwy oznaczono jako II stadium gza – Hypoderma bovis. Występowanie larw gzów z tego rodzaju u koni może zmniejszać ich wykorzystanie w jeździectwie.
The aim of these investigations was to prepare the antigen and control sera for the ELISA. Positive control sera were obtained from 163 cows showing clinical symptoms of H. bovis infection and negative control sera came from 60 cows free from the infection. The antigen for the ELISA was prepared from H. bovis L1 larvae. Three basic fractions were isolated from whole excretory/secretory antigen by elution from gel and by preparative isoelectrofocusing. Using the Western-Blotting method it was demonstrated that Hy A (22-24 kDa and pI 4-5) had the best antigenic property. As can be seen from these investigations, the developed ELISA antigen for the detection of H. bovis antibodies in cows is highly specific and sensitive.
A specimen of an unidentified parasite was taken out of the eye ball of an 8-year-old boy hospitalized in April 1964. A detailed analysis of the parasite showed that in was the Ist larval stage of Hypoderma bovis. The conservative treatment was unsuccessful and the eye ball, largely domaged by the parasite, had to be removed.
DNA barcoding is a practical tool for species identification, when morphological classification of an organism is difficult. Herein we describe the utilisation of this technique in a case of ophthalmomyiasis interna. A 12-year-old boy was infested during a summer holiday in northern Norway, while visiting an area populated with reindeer. Following medical examination, a Diptera larva was surgically removed from the boy’s eye and tentatively identified from its morphological traits as Hypoderma tarandi (L.) (Diptera: Oestridae). Ultimately, DNA barcoding confirmed this impression. The larval cytochrome c oxidase subunit 1 (COI) DNA sequence was matched with both profiles of five adult H. tarandi from the same region where the boy was infested, and other established profiles of H. tarandi in the Barcode of Life Data Systems (BOLD) identification engine.
The stability of 2 H. bovis antigens prepared by different methods was examined by the ELISA. It was found that the activity of H. bovis secretory antigen kept at - 18°C and -70° C and full antigen stored at - 18°C was significantly decreased as early as 6 months after storage. Lyophilized, secretory H. bovis antigen showed constant, high activity and may be used for one year at least.
The aim of the investigations was to determine the stability of ELISA components preserved under different conditions. The following components for ELISA were used: the secretory-excretory (s-e) antigen prepared from H. bovis L1 larvae, its fraction containing hypodermin A, conjugate, positive and negative control sera. Freezing at -20°C, lyophilization and preservation at 4°C were applied. Seven ELISA kits containing components preserved by the use of the described methods were prepared. The identical packets of sera were examined by ELISA directly after preparation of the kits (day „0”) and next at 3-4-week intervals for 9 months. The experiment showed that the best method for conserving ELISA components during the detection of H. bovis antibodies was lyophilization. The kits prepared in this way did not lose their characteristics for 9 months and their specificity and sensitivity were nearly 100%.
The aim of these investigations was to prepare the antigen and control sera for the ELISA. Positive control sera were obtained from 163 cows showing clinical symptoms of H. bovis infection and negative control sera from 60 cows free from the infection.The antigen for theELISA was obtained from H. bovis L1 larvae. Three basie fraction were isolated from whole excreted/secreted antigen by elution from gel and by preparative isoelectrofocusing.Using the Western-Blotting method it was demonstrated that Hy A (22-24 kDa and pI 4-5) had the best antigenic property. As can be seen from these investigations the developed ELISA kit for the detection of H. bovis antibodies in cows is highly specific and sensitive.
The aim of investigations was to determine the stability of ELISA components preserved under different conditions. The following components for the ELISA were used: the secreted/excreted antigen prepared from H. bovis L1 larvae, its fraction containing hypodermine A, conjugat, positive and negative control sera. Freezing at -20°C, lyophilization and preservation at 4°C were applied. Seven kits for the ELISA containing components preserved using described methods, were prepared., The identical packets of sera were examined by the ELISA directly after preparation of the kits (day '0'), and next, at 3-4 week intervals for 9 months. The experiments showed that the best method of conservation of ELISA components for the detection of H. Bovis antibodies was lyophilization. The kits prepared in this way did not lose their characteristics for 9 months and their specificity and sensitivity were close 100%.
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