Browsing by Author "Ekin, I. H."

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Comparison of Culture and Pcr for the Detection of Brucella Melitensis in Blood and Lymphoid Tissues of Serologically Positive and Negative Slaughtered Sheep
(Blackwell Publishing, 2008) Ilhan, Z.; Aksakal, A.; Ekin, I. H.; Gulhan, T.; Solmaz, H.; Erdenlig, S.
Aims: To compare the culture and PCR methods for detection of Brucella melitensis in blood and lymphoid tissue samples obtained from slaughtered sheep (n = 162) testing positive/negative in serological tests (Rose Bengal test and serum agglutination test). Methods and Results: Of 162 sheep examined, 45 were positive and 117 negative in serological tests. A PCR assay based on a pair of Br. melitensis-specific primers was used to detect DNA in blood and lymphoid tissue. Brucella melitensis was isolated from 1.2% (2/162) and 17.2% (28/162) of the blood and lymphoid tissue samples respectively. Positive PCR products with a molecular size of 731 bp were obtained from 27.7% (45/162) of blood and 29.0% (47/162) of lymphoid tissue samples. Conclusions: The species-specific PCR assay detected a higher number of Br. melitensis DNA both from serologically positive (P < 0.01 in blood PCR, P < 0.001 in tissue PCR) and serologically negative (P < 0.001 in both blood PCR and tissue PCR) sheep compared with classical bacteriological culture methods. Significance and Impact of the Study: The results emphasize the importance of using more than one type of diagnostic technique for the detection of animals positive for brucellosis, especially with epidemiological purposes.
Comparison of Pcr Assay and Bacteriological Culture Method for the Detection of Brucella Melitensis in Stomach Content Samples of Aborted Sheep Fetuses
(M H Schaper Gmbh Co Kg, 2007) Ilhan, Z.; Solmaz, H.; Aksakal, A.; Gulhan, T.; Ekin, I. H.; Boynukara, B.
The aim of this study was to evaluate the polymerase chain reaction (PCR) assay for detection of Brucella melitensis in stomach content samples of aborted sheep fetuses and to compare its performance with bacteriological culture method. It was also aimed to determine the agreement between PCR and Rose Bengal plate test (RBPT). Materials were collected from aborted sheep from 109 different sheep flocks in the region of Van during the lambing seasons of 2004-2005 and 2005-2006. Stomach contents from 135 aborted sheep fetuses were examined by bacteriological culture and PCR, and 135 sera from these aborting ewes were tested by RBPT Identification and typing of Brucella strains were performed using standard classification test. B. melitensis biovar 3 was isolated from 26 (19.2 %) of foetal stomach contents. B. melitensis was detected by PCR in 29 (21.4 %) stomach content samples. Twenty five sera (18.5 %) from aborting ewes tested positive by RBPT The detection limit of B. melitensis 16 M strain by PCR was 1.7 x 10(3) cfu (colony forming units) /ml in spiked stomach contents. Diagnostic sensitivity and specificity of the PCR were detected as 100 % and 97.2 %, respectively. The agreement between PCR and RBPT was found to be 97 %. In conclusion, PCR assay would have an advantage over conventional bacteriological culture method, but in particular for its ability to meet the specificity requirements for the detection of B. melitensis in stomach content samples of aborted sheep fetuses.
Detection of Brucella Melitensis Dna in the Milk of Sheep After Abortion by Pcr Assay
(Univ Austral Chile, Fac Ciencias veterinarias, 2008) Ilhan, Z.; Solmaz, H.; Aksakal, A.; Gulhan, T.; Ekin, I. H.; Boynukara, B.
Laboratory diagnosis of brucellosis is generally performed by microbiological and serological methods. PCR assay is a specific and sensitive choice for the detection of different bacterial agents. An evaluation of this test was carried out for the detection of Brucella melitensis DNA in sheep milk. 102 milk samples from sheep after abortion were taken and studied using bacteriological culture, PCR and milk ring test (MRT). PCR found B. melitensis DNA in 24 (23.5%) out of 102 milk samples, while only 8 (7.8%) of the samples were positive to B. melitensis through direct culture. MRT found 28 (27.4%) positive milk samples. The detection limit for PCR in sheep milk inoculated with B. melitensis strain 16 M was 1.7 x 10(3) - 1.7 x 10(4) cfu/ml. PCR and MRT coincidence was 96%. The diagnostic sensitivity and specificity were determined as 100% and 81.3% respectively for PCR assay and 75% and 75% for MRT. PCR is a useful tool for a fast diagnosis of B. melitensis in sheep milk.
Determination of Campylobacter Fetus Subsp. Fetus and Campylobacter Jejuni in Aborted Sheep Fetuses by Multiplex Pcr Assay
(Israel veterinary Medical Assoc, 2021) Ilhan, Z.; Ekin, I. H.; Gulaydin, O.
Campylobacteriosis is a contagious and zoonotic infection characterized by abortion and infertility in animals. Ovine campylobacteriosis is caused by Campylobacter (C.) fetus subsp. fetus or C. jejuni. As a result of the slow-growing bacterial agents and the high lability of Campylobacter species, laboratory diagnosis of these agents has always been problematic. Several publications on detection of Campylobacter spp. DNA from reference strains or pure culture have been presented. However, studies that have been carried out on field or clinical samples of sheep are quite limited. The purpose of the current study was to evaluate the utilization of multiplex-PCR (m-PCR) assay in the diagnosis of ovine campylobacteriosis from abomasal content samples of aborted sheep fetuses. A total of 116 aborted sheep fetuses were tested and C. fetus subsp. fetus was isolated from 8 (6.9%) samples. The m-PCR assay was conducted and amplified C. fetus subsp. fetus-specific DNA in 13 (11.2%) of the abomasum contents. Cohen's kappa coefficient revealed substantial (kappa: 0.74) agreement between bacteriological culture and m-PCR methods. The results of this study suggest that the m-PCR assay is more sensitive and reliable than conventional bacteriological culture for detection of Campylobacter species.
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.
Sds-Page Analysis of Heat Shock Proteins of Enterococcus Faecalis and Enterococcus Faecium Isolates From Urine and Feces Samples
(Wiley, 2017) Ates, S.; Ekin, I. H.
Western Blot Analysis of the Igg-Antibody Response To Acid-Glycine Antigens From Campylobacter Fetus Subsp. Fetus and C-Jejuni in Naturally Infected Sheep
(veterinarni A Farmaceuticka Univerzita Brno, 2007) Gurturk, K.; Ekin, I. H.; Arslan, A.
IgG-antibody response in aborting sheep and in apparently healthy sheep in a flock against acid-glycine-extracted antigens from three strains for each C fetus subsp. fetus and C jejuni were analysed by Western blot. One strain of C fetus subsp. fetus was isolated from aborting sheep. Western blot analysis of the sera revealed the presence of IgG antibody binding to the common antigens including proteins with the Mw of 63 kDa and 54 kDa in extracts from both C. fetus subsp. fetus and C. jejuni strains. In addition, IgG antibodies in sera from aborting sheep reacted more strongly with the antigens from C. fetus subsp. fetus strains with Mw of approximately 100, 95 and 86.5 kDa than those of apparently healthy sheep. The binding profile of the antibodies with these antigens appeared to be unique for each C. fetus subsp. fetus strain. On the other hand, IgG antibodies only in sera from aborting sheep recognized strongly the antigens of each C. fetus subsp. fetus strain at the Mw ranged from approximately 26 to 22 kDa. However, the antigenic components between 26 and 22 kDa were not detectable in coomassie blue stained gel and thought to have non-protein nature. These low molecular weight antigens of C. fetus subsp. fetus may be related to a recent infection in aborting sheep. These observations indicate that such species-specific antigens or conjugated protein antigens could be used for improving the specificity of the serological tests to detect C. fetus antibodies in sheep sera, and may be the candidates for subunit vaccines against ovine abortion.