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    Detection of slime genes and antiseptic/antibiotic-resistance genes in Staphylococcal isolates from Damascus goats with subclinical mastitis
    (Ecole Nationale Veterinaire Toulouse, 2019) Cantekin, Z.; Ergun, Y.; Solmaz, H.; Tek, E.
    This study aimed to detect slime genes and antibiotic/antiseptic-resistance genes in staphylococcal isolates from goats with subclinical mastitis in Hatay, Turkey. Thirty staphylococcal isolates were subjected to Polymerase Chain Reaction (PCR) analyses for detection of the studied genes. The genes responsible for slime production (icaA and icaD), in addition to oxacillin (mecA), gentamicin (aacA-aphD), erythromycin (ermA and ermC), tetracycline (tetK and tetM) penicillin (blaZ) and quarterly ammonium compound (QAC) resistance genes (qacA/qacB and qacC) were analysed by PCR assays. Slime genes predominated in goat mastitis isolates (12/30, 40.0%). The most prevalent resistance gene was blaZ, which was found in 11 (36.6%) isolates. mecA was present in 2 (6.6%) isolates, and qacA/B was found in 1 (3.3%) isolate. The other resistance genes were not detected in any isolates. Twelve of the 30 isolates were negative for the studied genes. In conclusion, the presence of ica genes in 40% of isolates illustrates the slime production ability of staphylococcal isolates in goat mastitis. The presence of these resistance genes in goat mastitis isolates is remarkable and has implications for goat milk and goat milk products for human consumption.
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    Development of Polymerase Chain Reaction assays with host-specific internal controls for Chlamydophila abortus
    (Czech Academy Agricultural Sciences, 2015) Cantekin, Z.; Solmaz, H.; Ergun, Y.; Ozmen, M.
    Chlamydophila abortus (C. abortus) is one of the most important infectious agents causing abortion in ruminants. The bacterium is obligately intracellular, cannot grow on agar, but it needs cell culture or embryonated eggs for growth. Therefore, culture-independent detection methods such as the polymerase chain reaction are increasingly important and needed. The aim of this study was to develop a polymerase chain reaction assay with an internal control for detection of C. abortus in clinical samples. Using newly-designed two primer sets specific for C. abortus, the polymerase chain reaction assay was first tested with positive and negative control DNA and its sensitivity and specificity were determined. A new polymerase chain reaction protocol was developed by combining the new primer pair sets with bovine (12SM-FW and 12SBT-REV2) and ruminant host-specific primer sets (12S-FW and 12S-REV). In conclusion, the developed polymerase chain reaction assays can potentially be used for direct detection of Chlamydophila abortus in bovine and ruminant samples.
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    Distribution of antiseptic resistance genes in Staphylococcus spp. from bovine mastitis
    (Czech Academy Agricultural Sciences, 2017) Ergun, Y.; Cantekin, Z.; Gurturk, K.; Solmaz, H.; Ekin, I. H.; Ozturk, D.
    The purpose of this study was the determination of antiseptic resistance genes (qacA/B and qacC) from staphylococcal mastitis in cattle in various regions of Turkey. In total, 283 isolates (Burdur: 36, Hatay: 47 and Van: 200) were studied, and the antiseptic resistance genes were detected using simplex PCR. The distribution of the qacA/B and qacC genes, mediating resistance against quaternary ammonium compounds, was found to vary among the different isolates. The qacA/B genes were found in three of the Burdur isolates, six of the Hatay isolates and seven of the Van isolates. The qacC gene was found in two of the Burdur isolates, none of the Hatay isolates and two of the Van isolates. The presence of these genes and transmission among Staphylococcus spp. strains may pose risks in the control of mastitis, as well as to public health.
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    A PCR method with internal control for detection of Brucella spp. from bovine abortion samples
    (Ecole Nationale Veterinaire Toulouse, 2014) Solmaz, H.; Cantekin, Z.; Altug, N.; Ilhan, Z.; Aslan, S.; Ergun, Y.
    Brucella spp. are important infectious agents in bovine abortions worldwide. The bacteriological culture of Brucella spp. is fastidious and time consuming procedure as a classical laboratory method. Brucella spp. can be detected by using different molecular techniques. The aim of this study is to develop a PCR technique with an internal control for detection of Brucella spp. from bovine abortion samples. For this purpose, the sensitivities of three different primer pairs (BgF/BgR, B4/B5 and JP-R/JP-F) were compared. Bovine 12S gene specific primer pairs (12SM-FW/12SBT-REV2) were used as an internal control. The sensitivity of BgF/BgR primers was found higher than the other primer sets. A PCR assay was developed by combining BgF/BgR primer sets and primers for bovine 12S. This protocol was tested and validated by using abomasal contents of two Brucella-positive and eighteen Brucella-negative clinical samples. In conclusion, the developed PCR method with an internal positive control has a potential for use in direct detection and identification of the Brucella spp. from bovine abortion samples.