Usporedba vrijednosti dvaju različitih kompleta početnica za dokaz vrste Pasteurella caballi lančanom reakcijom polimerazom u uzorcima bronhoalveolarnog ispirka ždrebadi čistokrvnoga arapskog konja.

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 to be 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.Pasteurella caballi (P. caballi) izdvojena je i identificirana u uzorcima bronhoalveolarnog ispirka i tkiva pluća ždrebadi čistokrvnoga arapskoga konja uobičajenim mikrobiološkim postupcima. Potom je istražena vrijednost dvaju različitih kompleta početnica za dokaz i potvrdu vrste P. caballi lančanom reakcijom polimerazom. Početnica 1 za dokaz gena 16S rRNA P. caballi bila je pripremljena upotrebom programa Primer 3, a početnica 2 upotrebom programa Primer-BLAST. Lančana reakcija polimerazom rabljena je za potvrdu sojeva vrste P. caballi i za njezin izravni dokaz u uzorcima bronhoalveolarnog ispirka i plućnoga tkiva. Ukupno je bilo izdvojeno 35 izolata Pasteurella spp. iz 25 (38,4 %) od 65 pretraženih uzoraka bronhoalveolarnog ispirka i 10 (58,8 %) izolata iz 17 uzoraka plućnoga tkiva. Ti su sojevi bili identificirani na osnovi poznatih mikrobioloških i biokemijskih značajki. Početnice 1 i 2 pokazale su 100 %-tnu osjetljivost za identifikaciju uzgojenih sojeva. Međutim, početnica set-1 za dokaz vrste P. caballi bila je slabije specifičnosti od početnice set-2 pri pretrazi uzoraka bronhoalveolarnog ispirka i plućnoga tkiva. Osjetljivost i specifičnost početnice 2 bila je povrđena analizom genske sekvencije. Istraživanje pokazuje da početnica set-2 za lančanu reakciju polimerazom za dokaz 16S rRNA pruža brz i točan alat za dokazivanje i potvrdu izolata vrste P. caballi u bronhoalveolarnom ispirku i plućnom tkivu ždrebadi

Prevalence of chlamydophila psittaci infection in pigeons and paraquets by a real time polymerase chain reaction

Aim: Avian chlamydiosis is a systemic, zoonotic, sometimes fatal disease caused by Chlamydophila psittaci in domestic and wild poultry. Although C. psittaci does not always cause serious disease in poultry, it can cause serious disease especially in immunosuppressive humans. The aim of this study was to determine the prevalence of C. psittaci, in stool samples of pigeons and paraquets, in Konya province. Materials and Methods: Fifty pigeon feces samples taken from a total of 600 pigeons belonging to 24 different breeders and 52 fecal samples taken from 632 paraquets from 30 different breeders were the samples in this study. The presence of C. psittaci was investigated by Real Time PCR based on the presence of ompA gene in these samples. Results: The presence of C.psittaci ompA gene was determined as 2% and 1%, respectively, by real time PCR in stool samples of pigeons and paraquet in Konya region. Conclusion: The fact that the disease is zoonotic and people's close contact with poultry, especially pigeons and caged birds is increased the importance of the disease in terms of public healthday day by day

Determination of phenotypical and genotypical characterization and antimicrobial resistance genes of staphylococcus aureus isolated from milk of dairy cows with mastitis

Aim: Mastitis is one of the most common diseases of dairy cattle and causes significant economic losses. Staphylococcus aureus produces many virulence factors that facilitate the adhesion and penetration of damaged tissues, and thereby, cause subclinical mastitis. This study was aimed at investigating the phenotypic and genotypic characteristics and antimicrobial susceptibility of S. aureus.. Materials and Methods: A total of 241 S. aureus strains isolated from bovine mastitis cases were tested phenotypically (catalase, coagulase, haemolysis, DNase, mannitol fermentation and biofilm formation) and genotypically (by the polymerase chain reaction (PCR) technique). Antimicrobial susceptibility was tested using 15 different antibiotics. Results: While the isolates showed different levels of haemolytic activity (? 47%, ? 42%, ? 10% and ? 1%), only the ?-haemolytic strains produced a positive CAMP-like reaction. Although all isolates were able to grow on MSA containing 7.5% NaCl, mannitol fermentation activity was observed in 80.5% of the isolates. The nuc gene was detected in all isolates, but only 84.2% of the isolates showed DNase activity. The Congo red agar method can be used to detect the biofilm forming capability of isolates, but the crystal violet staining method gives more reliable results. The sec gene was the most common enterotoxin genes (84%). Three isolates harboured the mecA gene, but were sensitive to methicillin. Conclusion: Phenotypic variations among isolates result in the misclassification of S. aureus strains and require the use of molecular methods. The rapid and accurate molecular typing of S. aureus can aid in both determining the prevalence of this infectious microorganism and preventing epidemic infections

Evaluation of clinical efficacy of tilmicosin in the treatment of respiratory system infections of calves

Aim: The aim of this study was to evaluate the clinical efficacy of tilmicosin in the treatment of respiratory system infections of calves. Materials and Methods: This study was performed on 30 calves with respiratory system infections. Bronchoalveolar lavage (BAL) fluid samples were taken from the calves. Microbiological examinations of BAL fluid samples were performed and antibiotic susceptibilities of the agents were determined. A single dose of tilmicosin (10 mg/kg SC) was administered to each calves. Results: While bacterial growth was not observed in 11 of 30 BAL samples, bacterial growth of 19 was detected. Pasteurella multocida (P. multocida) was isolated in 14 of the sample. Remaining bacteria were 3 Trueperella pyogenes (T. pyogenes) and 2 Escherichia coli (E. coli). Amoxicillin, sefquinom, marbofloxacin, gamithromycin and tilmicosin were found to be effective antibiotics. Conclusion: As a result, the dominant bacterium in respiratory system infections is Pasteurella multocida and also tilmicosin was effective in the treatment of respiratory diseases of calves

Evaluation of the effectiveness of bacteriophage therapy against salmonella infections in mice

Aim: The increase of infections caused by antimicrobial resistant salmonellaehas become a serious problem worldwide. In this study, it was aimed to investigatethe efficacy of bacteriophage treatment in mouse models by preparinga cocktail with four ΦSP-3 lytic phages (Salmonella Dublin, S. Typhimurium, S.Anatum, S. Kentucky) isolated from cattle feces as an alternative application.Materials and Methods: A total of 80 mice (total 8 experimental groups andeach group included 10 mice) were challenged with salmonella strains by oralroute. After challenge, bacteriohage coctail to mice were administrated by oralroute. Mice were observed for occurence of morbidity and mortality for 20days. Also, faecal samples were bacteriologically examined to determine theeffect of bacteriophage treatment on the spreading of Salmonella species withfeces.Results: The morbidity and mortality were observed in two mice, administeredbacteriophage coctail following challenge with S. Dublin and S. Typhimurium.In addition, re-isolation of S. Dublin and S. Typhimurium from internalorgans in 2 death mice were done. The morbidity and mortality in mice challengedwith S. Kentucky and S. Anatum and administered bacteriophage coctailwas not observed and re-isolation from internal organs were not carried out.However, re-isolation from feces of mice in all groups were made.Conclusion: The findings of present study revealed that bacteriophage cocktailsobtained from cattle faeces prevented mortality and morbidity in Salmonellainfected mice, and reduced the spread of Salmonella spp. Therefore, bacteriophagetherapy could be used for protection against salmonella infections

Comparison of reverse-transcriptase polymerase chain reaction (rt-pcr) and rapid test for the detection of bovine rotavirus and bovine coronavirus in anatolian water

Aim: Coronaviruses and Rotaviruses are important virologic factors for both animal and human health in Turkey and the world. Bovine Rotavirus (BRV) and Bovine Coronavirus (BCoV) in cattle cause significant economic losses. The aim of this study was to determine the presence of BRV and BCoV in Anatolian buffaloes which were on the same farms with cattle. For this purpose, presence of these two viruses were investigated by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and BRV-BCoV Rapid tests and sensitivity and specificity ratios of these two tests were compared. Materials and Methods: In this study, 230 Anatolian buffaloes were clinically evaluated in cattle farms in Afyonkarahisar region. Fecal samples were collected from 27 buffaloes which had clinical signs (weakness, dehydration, vomiting, watery consistency and yellow stool). The fecal samples were evaluated by Rapid Test and RT-PCR for Bovine Rotavirus and Bovine Coronavirus. The analyzes were performed according to the procedure of the commercial RT-PCR and rapid kits. Results: The RT-PCR results were positive as 22.2% (6/27) for BRV and 3.7% (1/27 27/1) for BcoV while Rota-Corona Rapid test results were negative in all samples. When compared with RT-PCR results for both viruses, the rapid test sensitivity and specificity was determined as 0% and 100%, respectively. In addition, positive rates of BRV was statistically important as BCoV rate in analyzed samples (p Conclusion: In conclusion, low sensitivity of rapid test may be due to the change in the amount of virus scattered throughout the course of enteric infections

Production and development of vaccines for ornithobacterium rhinotracheale infection in turkeys

Aim: The purpose of this study was to prepare bivalent inactive Ornithobacterium rhinotracheale bacterin vaccines to measure the levels of antibodies against antigens in blood sera and to determine the efficacies of different O. rhinotracheale vaccines on turkeys. Materials and Methods: The bivalent inactivated O. rhinotracheale bacterin vaccines were prepared from O. rhinotracheale serotype A and B strains using aliminium hydroxide, mineral oil, aluminium hydroxide + ginseng and mineral oil + ginseng. After the sterility and the safety tests, laboratory efficiencies of vaccines (challange/protection and serological potency) were done on the turkeys (twice vaccinated with doses 0.25 ml and 0.5 ml at 5 and 8 weeks, respectively). Results: According to the challenge results, all the vaccines were found effective at 100%. Slide agglutination, micro serum agglutination and ELISA tests were used for the diagnosis. The vaccine containing mineral oil and ginseng as adjuvant induced significantly greater humoral immune response than others. Also, vaccine containing mineral oil and ginseng as adjuvant was determined to be more effective in the field trials in a company privately producing turkeys. Conclusion: O. rhinotracheale vaccines could be used for prevention of ornithobacteriosis in turkeys

Efficacies of corynebacterium pseudotuberculosis vaccines against caseous lymphadenitis in mice and sheep

Aim: This study was aimed to prepare Corynebacterium pseudotuberculosis vaccines against Caseous Lymphadenitis (CLA) and to determine the effectiveness of vaccines in mice and sheep.4 Materials and Methods: C. pseudotuberculosis vaccines were prepared in 3 different combination (PLD, bacterin and combined). Vaccines were subcutaneously administered twice (0.1 mL) 3 weeks intervals to mice. After vaccination at 15 days, mice were subcutaneously challenged with C. pseudotuberculosis and PLD of the challenge strain with LD50 dose. All mice were observed throughout 20 days for morbidity and mortality. Internal organs of deaths/euthanized mice were cultured for re-isolation of bacteria. Sheep were subcutaneously vaccinated twice (1 mL) with vaccines of commercial or combined, 3 weeks intervals. Blood sera were taken before and after vaccination at 45-60 days intervals. In addition, lymph nodes were examined microscopically by the presence any abscess. Antibody titers to C. psedotuberculosis in mice and sheep were determined by modified ELISA. Results: When compared to controls, antibody titers were significantly higher in vaccinated mice (P<0.05). In challenged trials with PLD, the ratio of morbidity and mortality was not observed in the vaccines groups PLD and combined. But, the lower ratio of morbidity and mortality in challenged trials with live bacteria were determined in the groups of bacterin and combined. When the levels of antibody in sheep were found to be high, abscess formation were lower in, comparison to controls. Conclusions: Especially combined C. pseudotuberculosis vaccines were concluded to be useful in prevention of CLA in sheep