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Comparison of two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples
2016
Budniak, Sylwia | Kędrak-Jabłońska, Agnieszka | Szczawińska, Anna | Reksa, Monika | Krupa, Marek | Szulowski, Krzysztof
Introduction: The aim of the study was to optimise and compare two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples including the liver, brain, and blood. Material and Methods: Three strains of L. monocytogenes and single strains of each of the species: L. ivanovii, L. innocua, L. grayi, L. welshimeri, and L. seeligeri were used. Additionally, five other species of bacterium were used to evaluate the specificity of the tests. Results: Specific amplification products were obtained for both multiplex PCR assays, which confirmed the tested strains as Listeria spp. and L. monocytogenes, respectively. Isolates of other species did not yield PCR products. Conclusion: Both multiplex PCR assays proved to be significantly sensitive and highly-specific methods for the detection of Listeria strains.
显示更多 [+] 显示较少 [-]Morphology and molecular study of Fascioloides magna – a growing threat to cervids (Cervidae) in Poland
2016
Houszka, Marek | Piekarska, Jolanta | Podkowik, Magdalena | Gorczykowski, Michał | Bania, Jacek
Introduction: The giant liver fluke, Fascioloides magna, has spread across Europe over the years posing a serious threat to the Polish cervid population. Material and Methods: Macroscopic and histopathological studies of the liver of 22 roe deer (Capreolus capreolus), 10 red deer (Cervus elaphus), and 6 fallow deer (Dama dama) were performed. Species determination of the recovered liver flukes and eggs was performed by PCR protocol amplifying fragments of ribosomal DNA (ITS2), according to a standard method. Results: The presence of F. magna was confirmed in three (13.6%) roe deer, seven (70.0%) red deer, and two (33.3%) fallow deer. The fluke eggs were found only in the stools of five red deer and one fallow deer. Conclusion: This study presents detailed pathological and histopathological changes in the liver of wild Polish cervids, including roe deer, which were subjected to such study for the first time. The hepatic lesions typical for different stages of liver cirrhosis varied depending on the host species and stage of the disease.
显示更多 [+] 显示较少 [-]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.
显示更多 [+] 显示较少 [-]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.
显示更多 [+] 显示较少 [-]Evaluation of hyaluronic acid, procollagen type III N-terminal peptide, and tissue inhibitor of matrix metalloproteinase-1 as serum markers of canine hepatic fibrosis
2016
Lidbury, Jonathan A. | Hoffmann, Aline Rodrigues | Fry, Joanna K. | Suchodolski, Jan S. | Steiner, Jörg M.
The only way to diagnose hepatic fibrosis in dogs is by histological assessment of a liver biopsy specimen. As this technique is invasive and susceptible to sampling variation, serum biomarkers are used to detect hepatic fibrosis in humans. The objective of this study was to assess the utility of hyaluronic acid (HA), procollagen type III N-terminal peptide (PIIINP), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) as serum markers of canine hepatic fibrosis. Serum samples were collected from 47 dogs with histologically confirmed hepatobiliary disease and 24 healthy dogs in order to measure concentrations of HA, PIIINP, and TIMP-1. Hepatic fibrosis was staged using a 5-point scoring scheme. There was no correlation between serum concentrations of HA or PIIINP and the severity of hepatic fibrosis. There was a negative correlation between serum concentration of TIMP-1 and the severity of hepatic fibrosis (r s = −0.33; P = 0.036). It was not possible to use serum concentrations of HA, PIIINP, or TIMP-1 to discriminate between dogs with absent-to-moderate hepatic fibrosis and those with marked-to-very-marked fibrosis. The results of this study do not support the utility of measuring serum concentrations of HA, PIIINP, or TIMP-1 for diagnosing canine hepatic fibrosis. Further studies are needed to support this finding.
显示更多 [+] 显示较少 [-]Pharmacokinetics and tissue elimination of flunixin in veal calves
2016
Kissell, Lindsey W. | Brinson, Patrick D. | Gehring, Ronette | Tell, Lisa A. | Wetzlich, Scott E. | Baynes, Ronald E. | Riviere, Jim E. | Smith, Geof W.
OBJECTIVE To describe plasma pharmacokinetic parameters and tissue elimination of flunixin in veal calves. ANIMALS 20 unweaned Holstein calves between 3 and 6 weeks old. PROCEDURES Each calf received flunixin (2.2 mg/kg, IV, q 24 h) for 3 days. Blood samples were collected from all calves before the first dose and at predetermined times after the first and last doses. Beginning 24 hours after injection of the last dose, 4 calves were euthanized each day for 5 days. Plasma and tissue samples were analyzed by ultraperformance liquid chromatography. Pharmacokinetic parameters were calculated by compartmental and noncompartmental methods. RESULTS Mean ± SD plasma flunixin elimination half-life, residence time, and clearance were 1.32 ± 0.94 hours, 12.54 ± 10.96 hours, and 64.6 ± 40.7 mL/h/kg, respectively. Mean hepatic and muscle flunixin concentrations decreased to below FDA-established tolerance limits (0.125 and 0.025 μg/mL, respectively) for adult cattle by 3 and 2 days, respectively, after injection of the last dose of flunixin. Detectable flunixin concentrations were present in both the liver and muscle for at least 5 days after injection of the last dose. CONCLUSIONS AND CLINICAL RELEVANCE The labeled slaughter withdrawal interval for flunixin in adult cattle is 4 days. Because administration of flunixin to veal calves represents extralabel drug use, any detectable flunixin concentrations in edible tissues are considered a violation. Results indicated that a slaughter withdrawal interval of several weeks may be necessary to ensure that violative tissue residues of flunixin are not detected in veal calves treated with that drug.
显示更多 [+] 显示较少 [-]Perfusion- and diffusion-weighted magnetic resonance imaging of the liver of healthy dogs
2016
Del Chicca, Francesca | Schwarz, Andrea | Grest, Paula | Kircher, Patrick R.
OBJECTIVE To describe the perfusion and diffusion characteristics of the liver in healthy dogs as determined by morphological, perfusion-weighted, and diffusion-weighted MRI. ANIMALS 11 healthy adult Beagles. PROCEDURES Each dog was anesthetized and underwent morphological, perfusion-weighted, and diffusion-weighted MRI of the cranial aspect of the abdomen. On the MRI images, a region of interest (ROI) was established for each of 6 structures (aorta, caudal vena cava, portal vein, hepatic parenchyma, splenic parenchyma, and skeletal [epaxial] muscle). The signal intensity was determined, and a time-intensity curve was generated for each ROI. The apparent diffusion coefficient (ADC) was calculated for the hepatic and splenic parenchyma in diffusion-weighted MRI images, and the normalized ADC for the liver was calculated as the ratio of the ADC for the hepatic parenchyma to the ADC for the splenic parenchyma. Dogs also underwent abdominal ultrasonography, and ultrasound-guided fine-needle aspirate samples and biopsy specimens were obtained from the liver for cytologic and histologic examination. RESULTS Cytologic and histologic results suggested that the liver was clinically normal in all dogs. Perfusion-weighted MRI parameters varied among the 6 ROIs. The mean ± SD ADC of the hepatic parenchyma was 0.84 × 10(−3) mm2/s ± 0.17 × 10(−3) mm2/s, and the mean normalized ADC for the liver was 1.8 ± 0.4. CONCLUSIONS AND CLINICAL RELEVANCE Results provided preliminary baseline information about the diffusion and perfusion characteristics of the liver in healthy dogs. Additional studies on dogs of various breeds with and without hepatopathies are necessary to validate and refine these findings.
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