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Wyświetlanie 1-2 z 2
Tytuł:
Prevalence of Toxoplasma gondii infection in cats in southwestern Poland
Autorzy:
Sroka, J.
Karamon, J.
Dutkiewicz, J.
Wójcik-Fatla, A.
Zając, V.
Cencek, T.
Powiązania:
https://bibliotekanauki.pl/articles/2081746.pdf
Data publikacji:
2018
Wydawca:
Instytut Medycyny Wsi
Tematy:
Polska
PCR
seroprevalence
Toxoplasma gondii
IFAT
cats
oocysts
coproscopy
Opis:
Objective. An assessment of the prevalence of Toxoplasma gondii infection in cats from southwestern Poland using serology, coproscopy and PCR methods. Materials and method. In total, 208 cats (139 females and 68 males), aged 0.5–12 years (mean=2.6) from 25 localities in southwestern Poland were examined by indirect immunofluorescence assay (IFAT) to estimate the T. gondii serological status. Faecal samples of 41 cats were examined for the presence of oocysts/DNA T. gondii by microscopy and Real-time/nested PCR. After flotation (with NaNO3), pellets from faecal samples were disrupted by 10 cycles of freezing (liquid nitrogen) and warming. DNA was extracted using QIamp DNA Stool Mini Kit (Qiagen), according to the manufacturer’s instruction. Results. The positive results in IFAT for anti-T. gondii IgG and IgM antibodies were found in 143 of 208 tested cats (68.8%). Among positive results, 14.5%, 34.1% and 51.4% were detected in titre ranges 128–512, 1,000–2,000 and ≥ 4,000, respectively. In 23.1% of cat sera anti- T. gondii IgM antibodies were found. The prevalence of anti-Toxoplasma antibodies was significantly greater in older cats (>1 year) (83.5%) than in younger cats (48.3%) (P<0.05), in females (74.1%) than in males (58.8%) (P<0.05), and in cats kept outdoors than indoors (69.7% vs. 16.7%) (P<0.01). Among the 41 faecal samples examined, the presence of structures resembling T. gondii oocysts was found in 2 samples (4.9%), and for one of these samples (2.4% of the total) the result was also confirmed by PCR. Conclusions. T. gondii infection in domestic cats is highly prevalent in southwestern Poland. Information on the prevalence of infection in cats can be useful for assessing T. gondii environmental contamination and the risk for public health.
Źródło:
Annals of Agricultural and Environmental Medicine; 2018, 25, 3; 1-5
1232-1966
Pojawia się w:
Annals of Agricultural and Environmental Medicine
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Optimization of flotation, DNA extraction and PCR methods for detection of Toxoplasma gondii oocysts in cat faeces
Autorzy:
Sroka, J.
Karamon, J.
Dutkiewicz, J.
Wójcik-Fatla, A.
Cencek, T.
Powiązania:
https://bibliotekanauki.pl/articles/2081971.pdf
Data publikacji:
2018
Wydawca:
Instytut Medycyny Wsi
Tematy:
flotation
Toxoplasma gondii
faeces
Real time PCR
nested PCR
cats
oocysts detection
Opis:
Introduction and objective. The aim of the study was to compare the effectiveness of selected oocysts concentration methods, DNA extraction protocols and PCR assays targeting the B1 gene, for the development of procedures which would be effective and useful in laboratory practice for the detection of T. gondii in faecal samples from cats. Materials and method. In order to compare the influence of the flotation fluids on microscopy and PCR detection of T. gondii, saturated solutions of saccharose, MgSO4, ZnSO4 and NaNO3 were used. To determine the sensitivity of PCR tests used: Real time PCR (RT) and nested PCR, water samples spiked with T. gondii tachyzoites and oocysts were tested. DNA was extracted using DNeasy Blood and Tissue Kit (Qiagen) (K1). The same PCR tests were used to assess the efficacy of T. gondii DNA detection in samples of cat faeces spiked with oocysts, using DNA extraction by a K1 set and a K2 set (QIamp DNA Stool Mini Kit, (Qiagen). Results. The initial results showed that NaNO3 was most useful as a flotation fluid due to the lack negative effect on the oocysts and amplification efficacy in PCR. The level of detection for water samples (100 μl) was determined as 100 tachyzoites and 1–50 oocysts in RT, and 2–20 oocysts in nested PCR. The limit of detection (LD) for stool samples (250 mg) spiked with oocysts, where the K1 set was used, determined as 250 and 5 oocysts in RT and nested PCR, respectively. For samples extracted with the K2 set, LD in RT was determined as 1–50 oocysts (depending on the variant) and 50 oocysts in nested PCR. Conclusions. The most effective methods for detection of T. gondii in cat faeces seem to be centrifugal flotation with NaNO3, followed by DNA extraction with removing of inhibitors (K2 set) and Real Time PCR targeting B1 gene.
Źródło:
Annals of Agricultural and Environmental Medicine; 2018, 25, 4; 680-685
1232-1966
Pojawia się w:
Annals of Agricultural and Environmental Medicine
Dostawca treści:
Biblioteka Nauki
Artykuł
    Wyświetlanie 1-2 z 2

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