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    Cryptosporidiosis outbreak on a dairy farm: Detection of Cryptosporidium parvum as a causative agent in the water source
    (Polska Akad Nauk, Polish Acad Sciences, Univ Warmia & Mazury Olsztyn, 2021) Karakavuk, M.; Can, H.; Doskaya, M.; Karakavuk, T.; Erkunt-Alak, S.; Koseoglu, A. E.; Gul, A.
    Diarrhea caused by parasitic agents is common in neonatal calves and leads to significant economic losses in cattle farms worldwide. Cryptosporidium spp. is one of the most frequently detected parasitic agents causing diarrhea in neonatal calves. The aim of this study was to investigate the presence of Cryptosporidium spp. on a dairy farm which a has major diarrhea problem. Samples were collected from calves, cows, drinking bowls, and two different artesian water sources, as well as from the environment. All fecal samples were investigated using Kinyoun acid-fast stained slides and real-time PCR targeting the Cryptosporidium spp. COWP gene. In addition, species identification was performed by nested PCR targeting the Cryptosporidium spp. COWP gene and sequencing. Cryptosporidium spp. was detected in 11 calves (30.55%; 11/36) by real-time PCR and the cows were negative. Among real-time PCR positive samples, only five were also found positive by microscopy. Moreover, Cryptosporidium spp. was found in one of the two artesian water sources and five environmental samples by real-time PCR. Among these positive samples, eight were sequenced. According to the RFLP pattern, BLAST and, phylogenetic analyses, all sequenced samples were Cryptosporidium parvum. These findings show the importance of C. parvum as a cause of calf diarrhea on dairy farms.
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    A preliminary study to develop a lateral flow assay using recombinant GRA1 protein for the diagnosis of toxoplasmosis in stray cats
    (Elsevier Ltd, 2023) Değirmenci, Döşkaya, A.; Can, H.; Gül, A.; Karakavuk, T.; Güvendi, M.; Karakavuk, M.; Gül, C.
    Toxoplasma gondii is a protozoan parasite that may infect many mammals including humans. Cats are one of the main sources of infection for humans. Therefore, routine screening of cats with tests that are inexpensive, rapid, and do not require sophisticated laboratory equipment is important. In this study, a lateral flow assay (LFA) was designed to rapidly diagnose toxoplasmosis in cats. For this purpose, we selected GRA1 protein of T. gondii due to its high antigenicity in diagnostic and vaccine studies. We further analyzed the immunological properties of GRA1 protein using in silico tools. Then, we expressed and purified recombinant GRA1 (rGRA1) protein and used it during the development of LFA to detect toxoplasmosis in serum samples (n = 40) of cats. According to the results, rGRA1 protein has negative GRAVY value, high aliphatic index, alpha helix, random coil and 12 B cell epitopes. The in silico data supported the high antigenic properties of rGRA1 protein and showed that it can be a good antigen candidate for LFA. Among 30 cat positive serum samples, 27 were found positive by the LFA while seronegative sera (n = 10) were negative by the LFA. The preliminary data showed that the LFA has high sensitivity (90 %) and specificity (100 %). When we used high responsive cat sera (i.e. sera that have optical density > 0.5 with ELISA) the sensitivity value reached 100 %. These results showed that rGRA1 protein is a good candidate to develop a LFA for rapid diagnosis of toxoplasmosis in cats. © 2023 Elsevier Ltd