Browsing by Author "Solmaz, 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 Avian Influenza a Viruses in Some Avian Species in Van Lake Basin by Real Time-Pcr, Isolation and Subtyping
(2011) Boynukara, B.; Gulhan, T.; Adizel, O.; Ilhan, Z.; Aksakal, A.; Durmus, A.; Solmaz, H.
In this study, feces samples collected during 37 months from February 2006 to March 2009 from 2013 animals consisting of 47 avian species covering irregular vagrant, transit migrant, winter visitor, migratory and native birds in the Van Lake Basin Turkey were tested by Real-Time PCR (RT-PCR) with respect to Avian Influenza (AI) type A virus M2 gene. Of them, 59 samples (2.9%) were found to be positive. RT-PCR positive samples were examined with the same method with respect to H5N1 and 4 samples (6.8%) were found to be positive. RT-PCR positive 59 samples were inoculated in Embryonated Chicken Egg (ECE) and AI type A virus was isolated from 12 samples (20.3%). Of the isolates, 3 were typed as H1N7, 2 as H7N9, 2 as H11N9 and 1 as H8N4 with Hemagglutination Inhibition (HI) and Neuraminidase Inhibition (NI) tests. About 4 isolates obtained from winter visitor Anas cylpeata which had been determined as H5N1 by RT-PCR and agarose gel electrophoresis, gave positive reaction by HI test both with HI and H5 antisera and all were typed as Nl by Nl-test. Feces samples found to be positive by RT-PCR belonged to avian species Anseriformes, Charadriiformes, Pas seriformes, Gruiformes and Phoenicopteriformes orders. The highest positivity was determined in winter visitor Anas acuta (37.1%) and Anas penelope (22.5%) ducks. Of the RT-PCR positive 59 samples, 43 (72.9%) were determined in the samples collected during winter and spring of 2006-2009. Positivity was determined at a rate of 35.2% in respect of AI type A by RT-PCR in different species sharing the same time and place. With this study, the presence of AI type A viruses in various wild birds in the Van Lake Basin was determined for the first time in Turkey. © Medwell Journals, 2011.
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.
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.
Evaluation of R Mutant Escherichia Coli J5 Bacterin Vaccine Against Coliform Mastitis in Cows
(2009) Solmaz, H.; Llhan, Z.; Tasal, I.; Aksakal, A.
This study has shown that vaccination of cows with E. coli J5 bacterin during the dry period and after calving reduced the incidence of naturally occurring clinical coliform mastitis during early lactation.
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.