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Application of FTA® Cards for detection and storage of avian influenza virus
2016
Jóźwiak, Michał | Wyrostek, Krzysztof | Domańska-Blicharz, Katarzyna | Olszewska-Tomczyk, Monika | Śmietanka, Krzysztof | Minta, Zenon
Introduction: The aim of the study was to test the utility of Flinders Technology Associates filter paper (FTA® Cards) for molecular detection and storage of avian influenza virus (AIV). Material and Methods: There were two strains of AIV used in the study: low pathogenicity H7N1 and high pathogenicity H5N1 subtypes. Detection of viral material was conducted using molecular RT-PCR and rRT- PCR method. Results: The infectivity of LPAIV/H7N1 and HPAIV/H5N1 was completely inactivated within 1 h and 24 h after adsorption to FTA® Cards at room temperature, respectively. Viruses stored on FTA® Cards had detection limit approximately 1 log₁₀ lower than live viruses. Viral RNA of both strains were detectable on the cards by rRT-PCR for a minimum of 150 d, irrespectively of storage temperatures (room temperature, -20ºC). RNA was also detected in all samples obtained from SPF chickens experimentally infected with HPAI/H5N1 on 3ʳᵈ and 4ᵗʰ day post-infection (p.i.). Conclusion: FTA® Cards enable safe and effective alternative transport of samples for molecular diagnosis of AIV.
اظهر المزيد [+] اقل [-]Effect of doxycycline concentrations in chicken tissues as a consequence of permanent exposure to enrofloxacin traces in drinking water
2016
Gbylik-Sikorska, Małgorzata | Posyniak, Andrzej | Śniegocki, Tomasz | Sell, Bartosz | Gajda, Anna | Tomczyk, Grzegorz | Żmudzki, Jan
Introduction: The main problem in poultry farming is the difficulty in producing food of animal origin without using antibacterial agents. Because most antibacterial compounds are dispensed in water, some water supply systems can be contaminated by antibiotics which are then administered to the animals unintentionally. This can lead to unexpected increases in antibiotic residues in food of animal origin. The aim of the present study was to determine whether the constant exposure of chicken broilers to enrofloxacin affects the withdrawal time of a therapeutic doxycycline that is intentionally administered to the chickens.Material and Methods: The concentrations of doxycycline, enrofloxacin, and ciprofloxacin were determined by LC-MS/MS in muscles and liver of the chickens.Results: Doxycycline residue concentrations in the chicken tissues from the group that received trace amounts of enrofloxacin were nearly 50% greater than those of the group that received only doxycycline.Conclusion: These results indicated that constant exposure to enrofloxacin in trace amounts significantly influences the residual doxycycline concentration in chicken tissues.
اظهر المزيد [+] اقل [-]Pathogenicity of FtsK mutant of avian pathogenic Escherichia coli
2016
Xu, Xiaojing | Chen, Xiang | Gao, Song | Zhao, Lixiang
Introduction: Avian pathogenic Escherichia coli (APEC) is a leading cause of extraintestinal infection and heavy economic losses. Imparting immunity after vaccination with live attenuated strain vaccination is an ideal strategy for infection control. This study considers an FtsK knockout mutant strain as a candidate. Material and Methods: An FtsK knockout mutant of APEC strain E058 was constructed and the pathogenicity of the mutant and wild-type strains was further evaluated in chickens. Results: The 50% lethal doses of each strain for one-day-old specific-pathogen-free (SPF) chickens challenged experimentally via trachea were 10⁵.⁵ and 10⁷.⁰ colony-forming units (CFU) respectively. Chickens challenged with the wild-type strain exhibited typical signs and lesions of avian colibacillosis, while those inoculated with the mutant strain showed mild pericarditis and pulmonary congestion. The growth rate of the FtsK mutant strain was much slower than the wild-type strain in the heart, spleen, liver, and lung of infected chickens. Conclusion: These results indicated that the APEC FtsK mutant can be attenuated for chickens, and that this mutant has the potential for the development of an APEC vaccine.
اظهر المزيد [+] اقل [-]Development of a double-monoclonal antibody sandwich ELISA: Tool for chicken interferon-γ detection ex vivo
2016
Dai, Hua | Xu, Zheng-zhong | Wang, Meiling | Chen, Jun-hua | Chen, Xiang | Pan, Zhi-ming | Jiao, Xin-an
The aim of the present work was to develop reagents to set up a chicken interferon-γ (ChIFN-γ) assay. Four monoclonal antibodies (mAbs) specific for ChIFN-γ were generated to establish sandwich ELISA based on 2 different mAbs. To improve the detection sensitivity of ChIFN-γ, a double-monoclonal antibody sandwich ELISA was developed using mAb 3E5 as capture antibody and biotinylated mAb 3E3 as a detection reagent. The results revealed that this ELISA has high sensitivity, allowing for the detection of 125 to 500 pg/mL of recombinant ChIFN-γ, and also has an excellent capacity for detecting native ChIFN-γ. This ELISA was then used to detect ChIFN-γ level in chickens immunized with a Newcastle disease virus (NDV) vaccine, the immunized chicken splenocytes were stimulated by NDV F protein as recall antigen. From our results, it appears that the sensitivity range of this sandwich ELISA test is adequate to measure the ex vivo release of ChIFN-γ.
اظهر المزيد [+] اقل [-]Drug use and antimicrobial resistance among Escherichia coli and Enterococcus spp. isolates from chicken and turkey flocks slaughtered in Quebec, Canada
2016
Boulianne, M. (Martine) | Arsenault, Julie | Daignault, Danielle | Archambault, Marie | Letellier, Ann | Dutil, Lucie
An observational study was conducted of chicken and turkey flocks slaughtered at federal processing plants in the province of Quebec, Canada. The objectives were to estimate prevalence of drug use at hatchery and on farm and to identify antimicrobial resistance (AMR) in cecal Escherichia coli and Enterococcus spp. isolates and factors associated with AMR. Eighty-two chicken flocks and 59 turkey flocks were sampled. At the hatchery, the most used antimicrobial was ceftiofur in chickens (76% of flocks) and spectinomycin in turkeys (42% of flocks). Virginiamycin was the antimicrobial most frequently added to the feed in both chicken and turkey flocks. At least 1 E. coli isolate resistant to third-generation cephalosporins was present in all chicken flocks and in a third of turkey flocks. Resistance to tetracycline, streptomycin, and sulfisoxazole was detected in > 90% of flocks for E. coli isolates. Antimicrobial resistance (AMR) was observed to bacitracin, erythromycin, lincomycin, quinupristin-dalfopristin, and tetracycline in both chicken and turkey flocks for Enterococcus spp. isolates. No resistance to vancomycin was observed. The use of ceftiofur at hatchery was significantly associated with the proportion of ceftiofur-resistant E. coli isolates in chicken flocks. In turkey flocks, ceftiofur resistance was more frequent when turkeys were placed on litter previously used by chickens. Associations between drug use and resistance were observed with tetracycline (turkey) in E. coli isolates and with bacitracin (chicken and turkey), gentamicin (turkey), and tylosin (chicken) in Enterococcus spp. isolates. Further studies are needed to provide producers and veterinarians with alternative management practices and tools in order to reduce the use of antimicrobial feed additives in poultry.
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