Browsing by Author "Tutun, Hidayet"

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    Detection of SARS-CoV-2 using five primer sets
    (Ankara Üniversitesi, 2020-12-25) Karagöz, Alper; Tutun, Hidayet; Arslantaş, Tutku; Altıntaş, Özlem; Koçak, Nadir; Altıntaş, Nadir; Veteriner Fakültesi
    A novel coronavirus (SARS-CoV-2) outbreak, responsible for a pneumonia-associated respiratory disorder (COVID-19), has started in early December 2019 in Wuhan, China, and has rapidly spread around the world. Rapid and accurate diagnostic testing plays a crucial role in tackling the COVID-19 pandemic. In this study, it was aimed to compare 5 primer sets designed to amplify different regions for the detection of SARS-CoV-2 and to perform sequence analysis. Conventional RT-PCR was carried out using primers targeting different regions of the virus genome including ORF1ab, Envelope (E), RNA-dependent RNA polymerase (RdRp), Spike (S) and Nucleocapsid (N) genes for the diagnosis of COVID-19. DNA sequence of ORF1ab gene from each sample were compared with the DNA sequence data of SARS-CoV-2 stored in the GenBank and ORF1ab phylogenetic tree was constructed. The amplicon sizes of ORF1ab, S, E, N and RdRp genes were 588 bp, 440 bp, 145 bp, 323 bp and 196 bp, respectively. The SARS-CoV-2 RNA was detected from 74% of total samples from RdRp gene, 87% for N gene, 74% for S gene, 61% for E gene and 82% for ORF1ab region. The ORF1ab sequences of SARS-CoV-2 from 82 patients were had 100% identity to the sequence of Wuhan isolate and among themselves. The phylogenetic analysis revealed that all isolates formed a cluster. The results of this study suggest that the N region is the best for SARS-CoV-2 identification.
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    Determination of antibiotic susceptibility, ESBL genes and pulsed-field gel electrophoresis profiles of extended-spectrum β-lactamase-containing Escherichia coli isolates
    (Ankara Üniversitesi, 2019-09-09) Tutun, Hidayet; Karagöz, Alper; Altıntaş, Levent; Koçak, Nadir; Veteriner Fakültesi
    The purpose of this study was to determine the phenotypic antibiotic susceptibility patterns, extended-spectrum β-lactamase (ESBL) genes and genotypic profiles of ESBL-positive Escherichia coli strains isolated from urine samples obtained from outpatients with urinary tract infection in Turkey. A total of 120 E. coli strains during 2017, 2018, and 2019 (40 patients per year) were examined for antibiotic susceptibility patterns by disc diffusion method, for ESBL genes using PCR and sequencing and for molecular typing by pulsed-field gel electrophoresis (PFGE) method. The isolates were evaluated for their sensitivity to 21 different antibiotics. Four different antimicrobial resistance patterns were determined according to antibiotic susceptibility status of the isolates. The β-lactamase genes detected in the isolates were CTX-M-15 + OXA-1 (n= 14), CTX-M-15 (n= 24), TEM-1 + CTX-M-15 (n= 52), TEM-1 + SHV-12 (n=6), SHV-12 1 (n= 6), TEM-1 + CTX-M-1 (n= 6), TEM-1 + CTX-M-16 (n= 6) and TEM-1 + CTX-M-9 (n= 6). The CTX-M-15 was the most prevalent ESBL enzyme in the isolates. As a result of PFGE analysis performed by XbaI enzyme restriction process, one major PFGE profile and three main groups (Group I-II-III) were observed. While antibiotic resistance profiles of the strains showed four groups (RI-RII-RIII-RIV), PFGE band profiles showed a major group (90% similarity ratio). High ESBL production and decreased susceptibility to broad-spectrum cephalosporins were observed in E. coli strains. In addition, PFGE analysis showed high clonal similarity among E. coli isolates.
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    Effects of different chilling procedures on honey bees (Apis mellifera) for anesthesia
    (Ankara Üniversitesi, 2020-06-03) Tutun, Hidayet; Sevin, Sedat; Çetintav, Bekir; Veteriner Fakültesi
    Immobilization is required for the examination and manipulation of honey bees (Apis mellifera), and many techniques have been developed to render honey bees immobile until this date. Among them, two methods, which are carbon dioxide (CO2) and low-temperature narcosis, have been commonly used to induce anesthesia on the bees. Although CO2-induced anesthesia changes the behavior and physiology of honey bees, the anesthesia induced by cold is considered commonly as non-damaging for the bees. There are a very limited number of studies on the effects of different chilling protocols in honey bees. Therefore, the purpose of this study was to investigate the effect of different chilling procedures on the survival rate and depth of anesthesia in honey bees. A total of 4 groups were formed, one of which was the control group. Three temperatures (+4°C, -20°C and -80°C) were applied to three different experimental groups. Furthermore, four different exposure times were implemented on each experimental group (n=20 worker bees in each trial). The depth of anesthesia was assessed and scored based on the movements and anesthesia recovery time in honeybees. There was a statistically significant relationship between duration and survival/death rate in the applications at +4°C, -20°C and -80°C (P<0.001). The method that was applied for 5 minutes at -20°C was the most ideal chilling method (P<0.03) and showed the lowest mortality in addition to the high depth of anesthesia. This study may assist in selecting the safest and deepest anesthesia method required in any study on honey bees.
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    Phenotypic and molecular characterization of Salmonella Enteritidis isolates
    (Ankara Üniversitesi, 2021-03-31) Karagöz, Alper; Altıntaş, Levent; Arslantaş, Tutku; Tutun, Hidayet; Koçak, Nadir; Altıntaş, Özlem; Veteriner Fakültesi
    Salmonella spp. is the most frequently isolated foodborne pathogens causing human and animal diseases. The aim of this study was to investigate antimicrobial susceptibility profiles and the molecular typing of 200 Salmonella Enteritidis strains isolated from the patients’ stools between 2016 and 2019 in Turkey. The isolates were examined for antibiotic susceptibility patterns (21 antimicrobial agents) by Kirby-Bauer disc diffusion method or molecular typing by Pulsed-field gel electrophoresis (PFGE) and plasmid profiling. Although all isolates were susceptible to four antibiotics (suphamethoxazole/trimethoprim, chloramphenicol, streptomycin and trimethoprim), all were resistant to 15 different antibiotics. In the PFGE study performed with XbaI enzyme, all isolates were found to be related to each other according to similarity rates of 85% and above. There were two major clones, clone A and B. Clone A was divided into 6 pulsotypes (A1-A2-A3-A4-A5-A6) and clone B was divided into 3 pulsotypes (B1-B2-B3). Clone A had 87% similarity and Clone B had 90% similarity. The clustering rate was 86% (172/200). All isolates harboured 1-4 plasmid ranging in size from 2.5 to 57 kb and showed 6 plasmid profiles (P1-P6). All isolates carried the 57 kb plasmid individually or in combination with other plasmids. Most of the isolates 136 (68%) had P2 profile. Our findings indicate that the majority of all isolates were clonally related and had cross contamination problems. In this study, the importance of molecular typing methods in order to take more effective protection and control measures against Salmonella has been demonstrated and proposed to use such methods.
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    Sıçanlarda sipermetrin toksisitesine bağlı karaciğer hasarında CB2 agonisti JWH-133'ün sağaltıcı etkisinin araştırılması
    (Sağlık Bilimleri Enstitüsü, 2016) Tutun, Hidayet; Baydan, Emine; Other
    Sipermetrin tarımda, evde böcek kontrolünde besinlerin korunması ve hastalık vektör kontrolünde yaygın olarak kullanılan bir sentetik piretroid insektisittir. Bu çalışmada, sipermetrin toksisitesinde ksenobiyotiklerin uğrak yeri olan karaciğerde kannabinoid reseptör 2 (CB2) ekspresyonunun olup olmayacağının ve CB2 agonisti JWH-133'ün sipermetrinin toksik etkisini antagonize edip etmeyeceğinin belirlenmesi amaçlanmıştır. Araştırma, 44 adet Wistar Albino erkek şıçanda gerçekleştirilmiştir. Çalışma başlamadan 6 sıçan rastgele seçilerek kan ve karaciğer doku örnekleri alınmıştır. Geriye kalan sıçanlar 4 gruba (sırasıyla, kontrol; sipermetrin; sipermetrin + JWH-133; JWH-133) ayrılmıştır. Sipermetrin 14 gün boyunca 125 mg/kg ağızdan, JWH-133 sipermetrin toksisitesinin son 4 günü boyunca 3 mg/kg dozda intraperitoneal yolla verilmiştir. Çalışmanın sonunda, sıçanlardan kan (serumda AST, ALT, ALP and LDH) alındıktan sonra dekapite edilerek karaciğer dokularında oksidatif stres enzimleri, CB2 yönünden immunohistokimya ve RT-PCR ve histopatolojik inceleme yapılmıştır. Sonuçların değerlendirilmesinde SPSS 14.01 paket programı kullanıldı. Deneme grupları arasında CB2 mRNA miktarları bakımından fark bulunamamıştır. İmmunohistokimyal çalışmada CB2 reseptörleri normal karaciğer dokusunda çok zayıf gözlenmiştir. Ancak, sipermetrin uygulanan gruplarda CB2 reseptörlerinin expresyonunda artış gözlenmiştir. Artış karaciğer fibrojenik, Kuppfer, safra kanalı epitel ve mast hücrelerinde meydana gelmiştir. JWH-133, hem Grup 3 hem de Grup 4'te lipit peroksidasyonu düşürmüştür. Grup 3'te serum AST ve ALT enzim aktivitesi diğer gruplara göre artmıştır. Histopatolojik bakıda Grup 3 te Grup 2'ye göre parankim dejenerasyonu daha da güçlenmiştir. Sonuçlara göre sipermetrin toksisitesinde karaciğerde CB2 reseptörlerinin özellikle hepatik fibrojenik hücrelerde ekpresyonunun arttığı ve JWH-133 lipit peroksidasyonu düşürmesine rağmen karaciğer hasarını azaltamadığı aksine sipermetrinin karaciğer üzerine zararlı etkisini arttırdığı gözlenmiştir. Kannabinoid agonisti JWH-133'ün reseptör veya reseptör dışı yolaklar ile sipermetrinin toksik etkisini artırmış olabilir.