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Chlamydia psittaci reference genes for normalisation of expression data differ depending on the culture conditions and selected time points during the chlamydial replication cycle
2016
Van Lent Sarah | Vanrompay Daisy
Introduction: Chlamydia psittaci is a gram-negative obligate intracellular pathogen of birds. Poultry infections lead to economic losses and can be transmitted to humans. No vaccine is available and the bacterium-host cell interaction is not completely understood. Replicating bacteria cause pneumonia, but C. psittaci can also be non-replicating and persistent inside the cytoplasm of avian cells. RT-qPCR provides insight into the molecular pathogenesis of both active replicating and persistent Chlamydia psittaci in birds, but requires identification of stably expressed reference genes to avoid biases. Material and Methods: We investigated the expression stability of 10 C. psittaci candidate reference genes for gene expression analysis during normal growth and penicillin-induced persistence. C. psittaci Cal10 was cultured in HeLa229 and RNA was extracted. The expression level of each candidate was examined by RT-qPCR and Cq values were analysed using geNorm. Results: The genes tyrS, gidA, radA, and 16S rRNA ranked among the most stably expressed. The final selected reference genes differed according to the bacterial growth status (normal growth versus persistent status), and the time points selected during the duration of the normal chlamydial developmental cycle. Conclusion: The study data show the importance of systematic validation of reference genes to confirm their stability within the strains and under the conditions selected.
Afficher plus [+] Moins [-]Comparison of two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples
2016
Budniak Sylwia | Kędrak-Jabłońska Agnieszka | Szczawińska Anna | Reksa Monika | Krupa Marek | Szulowski Krzysztof
Introduction: The aim of the study was to optimise and compare two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples including the liver, brain, and blood. Material and Methods: Three strains of L. monocytogenes and single strains of each of the species: L. ivanovii, L. innocua, L. grayi, L. welshimeri, and L. seeligeri were used. Additionally, five other species of bacterium were used to evaluate the specificity of the tests. Results: Specific amplification products were obtained for both multiplex PCR assays, which confirmed the tested strains as Listeria spp. and L. monocytogenes, respectively. Isolates of other species did not yield PCR products. Conclusion: Both multiplex PCR assays proved to be significantly sensitive and highly-specific methods for the detection of Listeria strains.
Afficher plus [+] Moins [-]Red foxes (Vulpes vulpes) as reservoirs of respiratory capillariosis in Serbia
2016
Ilić Tamara | Becskei Zsolt | Tasić Aleksandar | Stepanović Predrag | Radisavljević Katarina | Đurić Boban | Dimitrijević Sanda
Introduction: The aim of the study was to determine the prevalence of respiratory capillariosis in red foxes (Vulpes vulpes) in some regions of Serbia.
Afficher plus [+] Moins [-]Investigation of human papillomaviruses (HPV), mouse mammary tumour virus (MMTV), Epstein–Barr virus (EBV), and human polyomavirus entities in canine mammary tumours
2016
Sonmez Kivilcim | Altan Eda | Yildirim Funda | Arun Seçkin Serdar | Turan Nuri | Yilmaz Huseyin | Kuskucu Mert Ahmet
Introduction: The aim of the study was to investigate the presence of human papillomaviruses (HPV), mouse mammary tumour virus (MMTV), Epstein-Barr virus (EBV), and human polyomavirus BK in canine mammary tumours (CMTs) and to correlate the results of histopathological classification with the results of virological examination. Material and Methods: Eighty CMTs and ten normal canine mammary gland samples were evaluated using histopathological methods and TaqMan real-time PCR analysis. Results: The results indicated that all mammary tumours and normal mammary tissue samples were negative for HPV16 and other HPV, EBV, human polyomavirus, and human mammary tumour virus strains. Conclusion: Further studies should be performed to investigate the existence of other strains of HPV, EBV, and human polyomavirus in CMTs.
Afficher plus [+] Moins [-]Application of FTA® Cards for detection and storage of avian influenza virus
2016
Jóźwiak Michał | Wyrostek Krzysztof | Domańska-Blicharz Katarzyna | Olszewska-Tomczyk Monika | Śmietanka Krzysztof | Minta Zenon
Introduction: The aim of the study was to test the utility of Flinders Technology Associates filter paper (FTA® Cards) for molecular detection and storage of avian influenza virus (AIV). Material and Methods: There were two strains of AIV used in the study: low pathogenicity H7N1 and high pathogenicity H5N1 subtypes. Detection of viral material was conducted using molecular RT-PCR and rRT- PCR method. Results: The infectivity of LPAIV/H7N1 and HPAIV/H5N1 was completely inactivated within 1 h and 24 h after adsorption to FTA® Cards at room temperature, respectively. Viruses stored on FTA® Cards had detection limit approximately 1 log10 lower than live viruses. Viral RNA of both strains were detectable on the cards by rRT-PCR for a minimum of 150 d, irrespectively of storage temperatures (room temperature, -20ºC). RNA was also detected in all samples obtained from SPF chickens experimentally infected with HPAI/H5N1 on 3rd and 4th day post-infection (p.i.).
Afficher plus [+] Moins [-]Impact of early castration and health status on the performance of Holstein-Friesian and beef crossbred calves
2016
Wielgosz-Groth Zofia | Sobczuk-Szul Monika | Nogalski Zenon | Purwin Cezary | Pogorzelska-Przybyłek Paulina | Winarski Rafał
Introduction: This article presents the analysis of the correlation between the category and health status of calves and the results of their rearing and levels of selected blood parameters.
Afficher plus [+] Moins [-]Identification of bacterial pathogens and determination of their antibacterial resistance profiles in some cultured fish in Turkey
2016
Türe Mustafa | Alp Hüseyin
Introduction: In the present study, some of the commercial fish farms located in the Black Sea region of Turkey, were screened for bacteria between 2012 and 2014.
Afficher plus [+] Moins [-]Changes in blood parameters induced by experimental jaagsiekte sheep retrovirus infection
2016
Dudek Katarzyna | Lutnicki Krzysztof | Bednarek Dariusz | Marczuk Jan | Kycko Anna | Reichert Michał
The aim of the study was to assess the changes of blood parameters in 12 three-week-old Polish Merino sheep subjected to experimental jaagsiekte sheep retrovirus (JSRV) infection.
Afficher plus [+] Moins [-]Usefulness of immunohistochemical indicators for diagnosis and prognosis of poorly differentiated tumours
2016
Kandefer-Gola Małgorzata | Nowak Marcin | Ciaputa Rafał | Madej Janusz A.
Immunohistochemical studies have become an indispensable element of establishing the correct histopathological diagnosis of poorly differentiated lesions, proving particularly suitable, and occasionally indispensable, for diagnosis of poorly differentiated neoplastic tumours. Knowledge of the mechanism of action and normal reaction of individual proteins is required in selection of the antibody pattern for a given tissue and in evaluation of the obtained results. This paper aims to promote the application of immunohistochemical techniques in routine diagnosis, especially in cases of poorly differentiated or undifferentiated tumours.
Afficher plus [+] Moins [-]Isolation and characterisation of peroxiredoxin 6 promoter from sheep (Ovis aries)
2016
Liu Nan-Nan | Liu Zeng-Shan | Lu Shi-Ying | Hu Pan | Zhang Ying | Fu Bao-Quan | Li Yan-Song | Zhou Yu | Zhang Yu | Ren Hong-Lin
Introduction: Peroxiredoxin 6 (Prdx6) is a bifunctional protein and a unique 1-Cys Prdx of the peroxiredoxin family. The expression and regulation of Prdx6 are implicated in numerous physiological and pathological processes.
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