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Öğe Antibacterial and Antibiofilm Effects of Boron on Different Bacteria(HUMANA PRESS INC, 2016) Sayin, Zafer; Ucan, Uckun Sait; Sakmanoglu, AsliBoron (B) compounds are used in many fields ranging from medicine to industry. In this study, boric acid (BA) and disodium octaborate tetrahydrate (DOT) were evaluated for their antibacterial effects and antibiofilm capacities on selected strains of clinical and type cultures that are of veterinary concern (Staphylococcus aureus ATCC 25923, Aeromonas hydrophila ATCC 19570, Pseudomonas aeruginosa ATCC 27853, Brucella melitensis Rev1 and field isolates of Vibrio anguillarum, Aeromonas hydrophila, Yersinia ruckeri, Pseudomonas aeruginosa, Lactococcus garvieae, and Brucella abortus). Also, the inhibition of biofilm was monitored by scanning electron microscopy. The lowest MIC values of BA and DOT were measured, by broth method using microdilution, from Pseudomonas aeruginosa ATCC 27853, and were 0.385 and 0.644 mg/ml, respectively. Staphylococcus aureus was the most resistant to both BA and DOT. Using the microplate method, we observed that the strongest positivities for biofilm production were presented by Pseudomonas aeruginosa ATCC 27853 and also a clinical isolate of Lactococcus garviea. Lower values in the MIC scores for both B compounds were tested by measuring the inhibitory effect on biofilm production. We found that all the bacterial strains inhibited biofilm formation with the exception of the Pseudomonas aeruginosa strains for BA only and an isolate of Lactococcus garviea for DOT only. Such effects by BA and DOT are worth discussing in order to find novel approaches for different functions in medicine and industry using the bacteria tested.Öğe Bacillus anthracis Isolated from a Dog in Turkey(UNIV AGRICULTURE, FAC VETERINARY SCIENCE, 2015) Sayin, Zafer; OzgurOzdemir; Erganis, Osman; Hadimli, Hasan Huseyin; Sakmanoglu, Asli; Atil, Eray; Sanioglu, GokcenurAn Anatolian Akbash shepherd dog died suddenly, without any clinical signs in Konya, Turkey. Performing necropsy, pathological examination, a culture test, laboratory tests and a multiplex-PCR of bacteria isolated from the dog revealed an anthrax and identified the bacteria as Bacillus anthracis (B. anthracis). The chromosomal gene sequence of bacteria was 99% identical in the GenBank under accession numbers CP002091 (B. anthracis str. H9401), CP001598 (B. anthracis str. A0248) and CP001215 (B. anthracis str. CDC684). Antimicrobial susceptibility test was performed and cefuroxime, cefquinome, sulfamethoxazole-trimethoprim, chloramphenicol, tetracycline and rifampicin resistance were seen in isolate. In this case, how the dog was infected with B. anthracis could not be determined. (C) 2014 PVJ. All rights reservedÖğe Comparison of the efficiency of concentrated soluble recombinant phospholipase D and natural phospholipase D enzymes(SCIENTIFIC TECHNICAL RESEARCH COUNCIL TURKEY-TUBITAK, 2016) Sakmanoglu, Asli; Erganis, OsmanPhospholipase D (PLD) is a major virulence determinant of Corynebacterium pseudotuberculosis. Various studies have focused on the expression of recombinant PLD (rPLD) enzyme in different Escherichia coli strains, but generally a high yield of insoluble protein was reported. The aims of this study were to express soluble rPLD by different methods in E. coli. The rPLD and natural PLD (dPLD) enzymes were concentrated using an ultramembrane cassette system after the efficiencies of these concentrated enzymes were compared. The rPLD enzyme was expressed in One Shot (R) BL21(DE3) E. coli when induced by IPTG in TY medium. Soluble dPLD and rPLD enzyme hemolytic activities were determined using the reverse CAMP test. The nucleotide sequence of the rPLD gene was 99.7% similar to the PLD gene of C. pseudotuberculosis in the NCBI GenBank Database; these differences in nucleotides resulted in a difference in two amino acids. The rPLD protein concentration and the titer of hemolytic activity were 23.1 mg/mL and 1/256, respectively. Similarities in the enzyme characteristics were detected between rPLD and dPLD enzymes. These findings indicate that a protocol would be useful for the enhanced production of soluble rPLD in E. coli and a membrane cassette system for concentration the recombinant protein.Öğe Comparison of two different primer sets for detection of Pasteurella caballi in bronchoalveolar lavage fluids samples from thoroughbred Arabian foals, using PCR(UNIV ZAGREB VET FACULTY, 2016) Sayin, Zafer; Erganis, Osman; Sakmanoglu, Asli; Hadimli, Hasan H.; Pinarkara, Yasemin; Maden, Mehmet; Al Shattrawi, Huda J. G.In the present study, Pasteurella caballi (P. caballi) was isolated and identified in bronchoalveolar lavage fluid and lung samples from thoroughbred Arabian foals using conventional microbiological methods. Subsequently, the ability of two different PCR primer sets was evaluated for detection and confirmation of P. caballi. Primer sets 1 and 2, targeting the 16S rRNA gene of P. caballi, were designed using the Primer 3 and Primer-BLAST programs, respectively. PCR was performed to confirm P. caballi strains and to detect it directly in the bronchoalveolar lavage fluid and lung samples. In total, 35 Pasteurella spp. were isolated from 25 (38.4 %) of 65 bronchoalveolar lavage fluid samples, and 10 (58.8 %) of 17 lung samples. These strains were identified as P. caballi based on conventional microbiological and biochemical characteristics. The sensitivities of primers 1 and 2 were determined lobe 100 % to confirm cultured P. caballi strains. However, the specificity of P. caballi detection was lower with primer set-1 than primer set-2 in bronchoalveolar lavage fluid and lung samples. The sensitivity and specificity of primer set-2 were confirmed by gene sequence analysis. This study indicates that the 16S rRNA-PCR method, using primer set-2, provides a rapid and accurate tool for the detection and confirmation of P. caballi isolates in bronchoalveolar lavage fluid and lung samples from foals.Öğe The Effectiveness of Anti-R. equi Hyperimmune Plasma against R. equi Challenge in Thoroughbred Arabian Foals of Mares Vaccinated with R-equi Vaccine(HINDAWI LTD, 2014) Erganis, Osman; Sayin, Zafer; Hadimli, Hasan Huseyin; Sakmanoglu, Asli; Pinarkara, Yasemin; Ozdemir, Ozgur; Maden, MehmetThis study aimed to determine the effectiveness of a pregnant mare immunization of a Rhodococcus equi (R. equi) vaccine candidate containing a water-based nanoparticle mineral oil adjuvanted (Montanide IMS 3012) inactive bacterin and virulence-associated protein A (VapA), as well as the administration of anti-R. equi hyperimmune (HI) plasma against R. equi challenge in the mares' foals. The efficacy of passive immunizations (colostral passive immunity by mare vaccination and artificial passive immunity by HI plasma administration) was evaluated based on clinical signs, complete blood count, blood gas analysis, serological response (ELISA), interleukin-4 (IL-4) and interferon gamma (IFN-gamma), total cell count of the bronchoalveolar lavage fluids (BALF) samples, reisolation rate of R. equi from BALF samples (CFU/mL), lung samples (CFU/gr), and lesion scores of the organs and tissue according to pathological findings after necropsy in the foals. The vaccination of pregnant mares and HI plasma administration in the foals reduced the severity of R. equi pneumonia and lesion scores of the organs and tissue by 3.54-fold compared to the control foals. This study thus indicates that immunization of pregnant mares with R. equi vaccine candidate and administration of HI plasma in mares' foals effectively protect foals against R. equi challenge.Öğe Efficacy of experimental inactivated and live Rhodococcus equi vaccines for thoroughbred Arabian mares in mice(SCIENTIFIC TECHNICAL RESEARCH COUNCIL TURKEY-TUBITAK, 2015) Erganis, Osman; Hadimli, Hasan Huseyin; Sayin, Zafer; Sakmanoglu, Asli; Pinarkara, Yasemin; Ozdemir, OzgurThe aim of this study was to determine the efficacy of inactive Rhodococcus equi vaccine candidates included that bacterin+aluminum hydroxide Al(OH)(3)), bacterin+VapA+(Al(OH)(3)), bacterin+Montanide IMS 3012 (IMS), bacterin+ VapA+IMS, and live vaccine using mice as a model. The efficacy of vaccine was evaluated according to clinical findings, humoral and cellular immunity (levels of INF-g and IL-4), and results of microbiological culture from internal organs in dead or sacrificed mice. Inactive R. equi vaccines were subcutaneously administered to mice three times at 15-day intervals and live vaccine was intraperitoneally injected once. Fifteen days after the last vaccination, aerosol challenges were carried out with the pathogenic R. equi VapA(+)K2002 strain in all groups. Two mice were sacrificed from each challenge groups on days 1, 3, 5, and 7. The antibody titers of vaccinated mice were found to be significantly higher than those of the controls. The largest number of INF-g positive samples were detected in the bacterin+ VapA+IMS and bacterin+ IMS groups. IL-4 positivity was determined only in live vaccine groups. The lowest reisolation rate of R. equi from internal organs was observed in the bacterin+VapA+IMS group. It was concluded that R. equi vaccines, and especially bacterin+VapA+IMS, are useful to protect mice against R. equi infection.Öğe Genotypic Characterization of Bordetella bronchiseptica Strains Isolated from Stray and Pet Dogs(UNIV AGRICULTURE, FAC VETERINARY SCIENCE, 2016) Sayin, Zafer; Sakmanoglu, Asli; Erganis, Osman; Ucan, Uckun Sait; Hadimli, Hasan Huseyin; Aras, Zeki; Sanioglu, GokcenurBordetella bronchiseptica (B. bronchiseptica) is the most important pathogen associated with kennel cough in dogs. The presence of B. bronchiseptica in pet dogs and shelter dogs with clinical respiratory disease was investigated in present study. The genetic relatedness among the strains was determined to evaluate the role of stray dogs in spread of B. bronchiseptica to pet dogs by detection of virulence genes such as filamentous hemagglutinin (fha), pertactin (prn) and dermonecrotic toxin (dnt). We also performed the random amplified polymorphic DNA (RAPD) assay. A total of 96 B. bronchiseptica were isolated from stray and pet dogs. The fha, prn and dnt virulence genes were detected in 86, 83.3 and 61.4% strains, respectively by polymerase chain reaction (PCR) techniques. The most common genotype from stray and pet dogs was fha+prn+dnt+ as detected in 37.5% and 11.4% of all the strains, respectively. The RAPD assay showed that 3 different patterns were obtained from 96 B. bronchiseptica strains. Sixty one (63.5%) of them were clustered in one main group and then further placed in another 2 sub-groups by RAPD assay. Genetic association was seen between the B. bronchiseptica strains from stray and pet dogs. In conclusion, this study revealed that B. bronchiseptica is present at a higher rate in stray dogs than pet dogs. Stray dogs might have a significant role in the transmission of B. bronchiseptica to pet dogs. (C)2016 PVJ. All rights reservedÖğe Mycoplasma infections in dairy cattle farms in Turkey(SCIENTIFIC TECHNICAL RESEARCH COUNCIL TURKEY-TUBITAK, 2016) Sayin, Zafer; Sakmanoglu, Asli; Ucan, Uckun Sait; Uslu, Ali; Hadimli, Hasan Huseyin; Aras, Zeki; Ozdemir, OzgurMycoplasmas cause the most severe and economically costly diseases of cattle throughout the world. In this study, Mycoplasma species were isolated from calves and cows with suspected mycoplasmosis in Holstein dairy cattle farms within 7 geographical regions of Turkey between May 2010 and December 2015. Mycoplasma infections were positive in 17 (80.9%) of 21 dairy cattle farms and the overall percentage was calculated as 32.1%. The highest isolation rate occurred in the Southeastern Anatolia Region (42.8%), and the lowest was observed in the Mediterranean Region (19.6%). In total, 172 Mycoplasma spp. were isolated from samples. Using PCR analysis, 149 (87.6%) isolates were identified as Mycoplasma bovis (M. bovis). Eleven (6.3%) isolates were identified as M. alkalescens, 2 (1.1%) were M. canis, and 10 (5.8%) were M. bovigenitalium. The isolation rate was found to be increasing annually. In conclusion, mycoplasmosis is a common problem in Holstein dairy cattle farms in Turkey, and M. bovis is the most frequently encountered cause of mycoplasma infections. The isolation rate seems to have increased in correlation with increased live cattle imports. Additionally, M. alkalescens and M. canis were isolated and identified in respiratory tract infections in cattle from Turkey for the first time.Öğe Serotypes of Salmonella isolated from feces of cattle, buffalo, and camel and sensitivities to antibiotics in Turkey(SCIENTIFIC TECHNICAL RESEARCH COUNCIL TURKEY-TUBITAK, 2017) Hadimli, Hasan Huseyin; Pinarkara, Yasemin; Sakmanoglu, Asli; Sayin, Zafer; Erganis, Osman; Uslu, Ali; Al-Shattrawi, Huda JihadIn this study, Salmonella strains from dairy cows, calves with diarrhea, and buffalo and camel feces were isolated and serotyped. There were 869 feces samples (437 calves, 287 dairy cows, 100 buffalo, and 45 camels) collected from 13 provinces and 21 farms. Preenrichment feces samples were added to Rappaport-Vassiliadis medium. Samples were then subcultured on xylose lysine tergitol-4 agar and plates. Suspected colonies of Salmonella were confirmed by latex agglutination test. The isolates of Salmonella spp. were serotyped at the Etlik Central Veterinary Control Institute. In total, 40 Salmonella spp. were isolated, including 33 from calves, 5 from dairy cows, 1 from a buffalo, and 1 from a camel. Salmonella strains were serotyped as S. Kentucky (n = 23), S. Muenchen (n = 5), S. Anatum (n = 4), S. Gaminare (n = 4), S. Typhimurium (n = 1), S. Muenster (n = 1), S. Enteritidis (n = 1), and S. Abony (n = 1). The Kirby-Bauer disk diffusion method was used for the determination of antibiotic susceptibilities of isolates against 15 antibiotics. Salmonella isolates were determined as resistant to multiple antibiotics. In conclusion, 8 different Salmonella serotypes were found in dairy cows, calves, buffalo, and camels. Because of the occurrence of multiresistant isolates, biosafety measures and pathogen control processes are advised for Salmonella-associated risks to public health.
