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Öğe Development of A Multiplex PCR Method for Direct Detection of Common Mastitis Pathogens in Bovine Milk Samples(KAFKAS UNIV, VETERINER FAKULTESI DERGISI, 2017) Kalin, Recep; Karahan, Murat; Acik, Mehmet Nuri; Tasdemir, Bulent; Cetinkaya, BurhanThe aim of this study was to evaluate a simple and rapid DNA extraction method combined with a multiplex polymerase chain reaction (mPCR) for the identification of the major mastitis pathogens (Staphylococcus aureus, Streptococcus agalactiae, Escherichia coli and Mycoplasma bovis) from milk samples. Of the 200 California Mastitis Test (CMT) positive milk samples, 45 (22.5%), 21 (10.5%) and 11 (5.5%) were detected as positive for the presence of S. aureus, S. agalactiae and E. coli by culture, respectively. In mPCR by DNA isolation method optimised here, S. aureus, S. agalactiae and E. coli were detected in 26.5% (53/200), 12% (24/200) and 6% (12/200) of the milk samples, respectively. The abovementioned agents were observed in similar proportions when the samples were analysed by a commercial DNA isolation kit. On the other hand, M. bovis was not detected in any of the milk samples by either culture or mPCR methods. A significant difference was determined between the results of culture and mPCRs (P< 0.001). Diagnostic sensitivity and specificity of the optimised mPCR were calculated as 100% and 89.2% respectively, when culture results were considered as reference. The results suggest that the mPCR assay employed in this study could be used as an alternative routine diagnostic method for rapid, sensitive, and specific simultaneous detection of major mastitis agents in bovine milk samples.Öğe Investigation of Virulence and Cytolethal Distending Toxin Genes in Campylobacter spp. Isolated from Sheep in Turkey(MARY ANN LIEBERT INC, 2013) Acik, Mehmet N.; Karahan, Murat; Ongor, Hasan; Cetinkaya, BurhanThe presence of virulence and cytolethal distending toxin (Cdt) genes was investigated in isolates of Campylobacter jejuni, C. coli, C. lanienae, and C. lari that originated from intestinal contents and gallbladders of clinically healthy sheep. These genes have important roles in the pathogenicity of campylobacters. A total of 363 Campylobacter isolates (221 C. jejuni, 135 C. coli, five C. lanienae, and two C. lari) were used in this study. The frequency of racR, dnaJ, ciaB, pldA, flaA, and cadF virulence genes in all the isolates were determined to be 34.4%, 30%, 24.8%, 30.9%, 95%, and 81.3%, respectively, while the virB11 virulence gene could not be detected in any isolates. CdtA, cdtB, and cdtC genes were detected in 54.5%, 55.9%, and 52.3% of the isolates, respectively. None of the virulence and toxin genes examined here were detected in a total of 19 Campylobacter isolates consisting of 10 C. jejuni and nine C. coli. This is the first study investigating the presence of virulence and toxin genes in a large number of Campylobacter species isolated from clinically healthy sheep by scanning a large area. In addition, this is the first report investigating the presence of virulence and toxin genes in sheep-originated C. lanienae and C. lari isolates.Öğe Investigation of Yersinia ruckeri Infection in Rainbow Trout (Oncorhynchus mykiss Walbaum 1792) Farms by Polymerase Chain Reaction (PCR) and Bacteriological Culture(KAFKAS UNIV, VETERINER FAKULTESI DERGISI, 2012) Seker, Engin; Karahan, Murat; Ispir, Unal; Cetinkaya, Burhan; Saglam, Naim; Sarieyyupoglu, MustafaThis study has been carried out to investigate the presence of Yersinia ruckeri, the causative agent of Enteric Red Mouth (ERM) disease, in rainbow trout fish breeding farms located in Elazig and Malatya provinces in Eastern Turkey. For this purpose, bacteriological culture and a specific polymerase chain reaction (PCR) have been compare with examine blood and intestine samples collected from adult and young rainbow trout fish as well as water samples collected from all the farms. Y. ruckeri was isolated and identified from eight of the 17 fish farms giving a proportion of 52.9% at fish farm. The disease was noted in five of the rainbow trout farms located in Elazig and three of the 11 rainbow trout farms located in Malatya. The agent was detected in five of the six young fish breeding farms. The isolation percentages of the causative agent at fish level were calculated as 25.7% (131/510) in adult and 31.7% (19/60) in young fish, giving an overall proportion of 26.3% (150/570). All the isolates have been successfully identified as Y ruckeri by species-specific PCR. No isolation could be made from water samples. It is therefore concluded that Y. ruckeri infection poses a significant threat to the fish farms in the study area and necessary precautions should urgently be taken in order to minimize economical losses due to ERM disease. A specific PCR can be utilized successfully in aquaculture for rapid identification of bacterial agents which will help to prevent spread of infectious diseases and will therefore contribute to the productivity of fishery sector.Öğe Investigation of yersinia ruckeri infection in rainbow trout (Oncorhynchus mykiss Walbaum 1792) farms by polymerase chain reaction (PCR) and bacteriological culture(Veteriner Fakultesi Dergisi, 2012) Şeker, Engin; Karahan, Murat; Ispir, Ünal; Çetinkaya, Burhan; Sa?lam, Naim; Sarieyyüpo?lu, MustafaThis study has been carried out to investigate the presence of Yersinia ruckeri, the causative agent of Enteric Red Mouth (ERM) disease, in rainbow trout fish breeding farms located in Elazig and Malatya provinces in Eastern Turkey. For this purpose, bacteriological culture and a specific polymerase chain reaction (PCR) have been compare with examine blood and intestine samples collected from adult and young rainbow trout fish as well as water samples collected from all the farms. Y. ruckeri was isolated and identified from eight of the 17 fish farms giving a proportion of 52.9% at fish farm. The disease was noted in five of the rainbow trout farms located in Elazig and three of the 11 rainbow trout farms located in Malatya. The agent was detected in five of the six young fish breeding farms. The isolation percentages of the causative agent at fish level were calculated as 25.7% (131/510) in adult and 31.7% (19/60) in young fish, giving an overall proportion of 26.3% (150/570). All the isolates have been successfully identified as Y. ruckeri by species-specific PCR. No isolation could be made from water samples. It is therefore concluded that Y. ruckeri infection poses a significant threat to the fish farms in the study area and necessary precautions should urgently be taken in order to minimize economical losses due to ERM disease. A specific PCR can be utilized successfully in aquaculture for rapid identification of bacterial agents which will help to prevent spread of infectious diseases and will therefore contribute to the productivity of fishery sector.Öğe Investigation on Antimicrobial Resistance Levels of Escherichia coli Strains Isolated from Bovine Fecal Samples and Comparison with Guidelines of the Clinical Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST)(2023) Kalın, Recep; Karagülle, Burcu; Açık, Mehmet Nuri; Karahan, Murat; Öztürk, Yasin; Çetinkaya, BurhanThis study was carried out to investigate the antibiotic resistance levels of Escherichia coli isolates of bovine origin and to compare the results with CLSI and EUCAST guideline values. For this purpose, 97 E. coli strains isolated from fecal samples of cattle in 12 different farms were tested against 32 antibiotics by using the disk diffusion method. The zone diameters of 13 antibiotics examined within the scope of the study were compared according to the CLSI and EUCAST 2020 guidelines, and their consistency levels were evaluated statistically. The highest resistance rates in E. coli isolates were determined against tetracycline (68%), streptomycin (63.9%), ampicillin (58.8%), and doxycycline (50.5%) antibiotics. On the other hand, the isolates were found to be highly susceptible to amikacin and cephalosporin group antibiotics. When CLSI and EUCAST guidelines were compared, it was found that there were statistically significant differences between the resistance rates of nitrofurantoin, gentamicin, and amikacin. Only 10 (10.3%) of the isolates were detected to be susceptible to all the antibiotics tested, whereas 17.5% were resistant to 10 or more antibiotics. The results of this study showed that E. coli isolates of bovine origin were highly resistant against antibiotics used in the field for a long period, especially the number of isolates with multiple antibiotic resistance was striking. It was concluded that due to substantial inconsistencies between the CLSI and EUCAST guidelines for some antibiotics such as amikacin, nitrofurantoin, and gentamicin, there is an urgent need to execute necessary updates in both guidelines.
