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Molecular characterizations of phosphoprotein of rabies virus circulating in Korea
2012
Kim, H.H., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea | Yang, D.K., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea | Jeon, J.K., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea | Cho, S.D., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea | Song, J.Y., Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang, Republic of Korea
Rabies is a major zoonotic disease that causes approximately 55,000 human deaths worldwide on an annual basis. The nucleocapsid protein and glycoprotein genes of the Korean rabies virus (RABV) have been subjected to molecular and phylogenetic analyses. Although the phosphoprotein (P) has several important functions in viral infection and pathogenicity, the genetic characterizations of the P of Korean RABV isolates have not yet been established. In the present study, we conducted genetic analyses of P genes of 24 RABV isolates circulating in the Republic of Korea (hereafter, Korea) from 2008 to 2011. This study revealed that the P genes of Korean RABVs are genetically similar to those of RABV strains of lyssavirus genotype I including V739 (dogs, Korea), NNV-RAB-H (humans, India), NeiMeng925 (raccoon dogs, China), and RU9.RD (raccoon dogs, Russia). Among Korean isolates, the RABV P genes showed low variability in the variable domains among Korean isolates; they had specific consensus sequences and amino acid substitutions capable of identifying geographic characteristics and retained specific sequences thought to be important for viral function. These results provide important genetic characteristics and epidemiological information pertaining to the P gene of the Korean RABV.
Afficher plus [+] Moins [-]An improved multiplex PCR for diagnosis and differentiation of Mycoplasma hyopneumoniae and Mycoplasma hyorhinis
2012
Barate, Abhijit K., Kangwon National University, Chuncheon, Republic of Korea | Lee, H.Y., Kangwon National University, Chuncheon, Republic of Korea | Jeong, H.W., Kangwon National University, Chuncheon, Republic of Korea | Truong, Lam Quang, Kangwon National University, Chuncheon, Republic of Korea | Joo, H.G., Jeju National University, Jeju, Republic of Korea | Hahn, T.W., Kangwon National University, Chuncheon, Republic of Korea
A multiplex PCR was developed for the simultaneous detection and differentiation of Mycoplasma (M.) hyopneumoniae and M. hyorhinis in clinical samples. Improved sensitivity is advantage of this technique over the previously reported multiplex assay. It was capable of detecting as little as 125 fg genomic DNA from M. hyopneumoniae and 62.5 fg genomic DNA from M. hyorhinis. Application of this multiplex PCR method to field isolates showed that M. hyopneumoniae and M. hyorhinis were present in 29% (107 of 370) of lung specimens and no mycoplasmas were detected in 56% (208 of 370) of the slaughtered pigs' lungs. At the farm level, M. hyopneumoniae and M. hyorhinis were detected in 34 of 36 (94.4%) randomly selected farms. We conclude that this assay would prove itself a value tool for monitoring these mycoplasmal infections and both M. hyopneumoniae and M. hyorhinis have been widely spread in swine herds of Korea.
Afficher plus [+] Moins [-]Effects of barley and barley bran contaminated with Fusarium spp. on the growth and feed efficiency of fattening and growing pigs
2012
Lee, W.S., Jeju National University, Jeju, Republic of Korea | Lee, H.J., National Institute of Animal Science, RDA, Cheonan, Republic of Korea | Ki, K.S., National Institute of Animal Science, RDA, Cheonan, Republic of Korea | Noh, H.G., Korea National College of Agriculture and Fisheries, Suwon, Republic of Korea | Kang, S.J., National Institute of Animal Science, RDA, Cheonan, Republic of Korea | Jung, Y.H., National Institute of Animal Science, RDA, Cheonan, Republic of Korea | Baek, K.S., National Institute of Animal Science, RDA, Cheonan, Republic of Korea | Hur, T.Y., National Institute of Animal Science, RDA, Cheonan, Republic of Korea
The present study was carried out to investigate the effect of barley and barley bran contaminated with Fusarium spp on growth performance and feed efficiency of fattening and growing pigs. In experiment 1, total 48 fattening Landrace pigs were used in a fattening trial for 71 days. Pigs weighing around 75 kg were allocated into different substitution groups containing 0, 10, 20 and 30% of barley contaminated Fusarium spp. In experiment 2, total 16 growing Landrace pigs were used in a growing trial for 45 days. Pigs weighing around 29.4 kg were allocated into different substitution groups containing 0, 5, 10 and 20% of barley bran contaminated Fusarium spp. Mycotoxin concentrations of barley and barley bran contaminated with 30% Fusarium spp were 0.452 and 1.049 ppm for deoxynivalenol, 8.125 and 17.646 ppm for nivalenol and 0.023 and 0.029 ppm for zearalenone, respectively. In experiment 1, no differences were found in weight gain and feed intake between control group (0%) and 10 or 20% substitution groups, but in 30% substitution group, weight gain and feed intake were significantly lower (p less than 0.05) than those in control group. After slaughtering, the extended haemorrhage of the fundus region in stomach was observed in 20 or 30% substitution groups. In experiment 2, weight gain and feed intake were not significantly different among treatment groups. After slaughtering of experimental pigs, the extended haemorrhage of the fundus region in stomach was observed in pigs fed diet with 20% substitution group. These results suggest that the feeding of diet with contaminated highly levels of Fusarium spp was negative effect on growth and feed efficiency in growing and fattening pig.
Afficher plus [+] Moins [-]Effects of a urolith prevention diet on urine compositions of glycosaminoglycans, Tamm-Horsfall glycoprotein, and nephrocalcin in cats with calcium oxalate urolithiasis
2012
Lulich, Jody P. | Osborne, Carl A. | Carvalho, Mauricio | Nakagawa, Yasushi
Objective: To evaluate urine concentrations of glycosaminoglycans, Tamm-Horsfall glycoprotein, and nephrocalcin in cats fed a diet formulated to prevent calcium oxalate uroliths. Animals: 10 cats with calcium oxalate urolithiasis. Procedures: In a previous study conducted in accordance with a balanced crossover design, cats were sequentially fed 2 diets (the diet each cat was consuming prior to urolith detection and a diet formulated to prevent calcium oxalate uroliths). Each diet was fed for 8 weeks. At the end of each 8-week period, a 72-hour urine sample was collected. Concentrations of glycosaminoglycans, Tamm-Horsfall glycoprotein, and the 4 isoforms of nephrocalcin in urine samples collected during that previous study were measured in the study reported here. Results: Diet had no effect on the quantity of Tamm-Horsfall glycoprotein and nephrocalcin in urine. However, the urine concentration of glycosaminoglycans was significantly higher during consumption of the urolith prevention diet. Conclusions and Clinical Relevance: Feeding a urolith prevention diet increased the urine concentration of glycosaminoglycans, which are glycoprotein inhibitors of growth and aggregation of calcium oxalate crystals.
Afficher plus [+] Moins [-]Effects of gemcitabine and gemcitabine in combination with carboplatin on five canine transitional cell carcinoma cell lines
2012
Objective: To evaluate in vitro effects of gemcitabine alone and in combination with carboplatin on canine transitional cell carcinoma (TCC) cell lines. Sample: In vitro cultures of 5 canine TCC cell lines. Procedures: Cells were treated with gemcitabine, carboplatin, or a combination of both at various concentrations. Cell proliferation was assessed via a fluorescence-based microplate cell proliferation assay. Cell cycle was evaluated via propidium iodide staining, and apoptosis was assessed by measurement of caspase 3 and 7 enzymatic activity. Synergy between gemcitabine and carboplatin was quantified via combination index analyses. Results: Treatment of 5 canine TCC cell lines with gemcitabine or carboplatin decreased cell proliferation, increased apoptosis, and induced cell cycle arrest. Cell cycle arrest and apoptosis were markedly increased when cell lines were treated with both gemcitabine and carboplatin simultaneously or sequentially. Order of administration during sequential treatment did not consistently affect cell proliferation results in TCC cell lines. When TCC cell lines were treated with gemcitabine and carboplatin in combination at therapeutically relevant concentrations (gemcitabine concentration, < 10μM; carboplatin concentration, < 250μM), a significant decrease in cell proliferation was observed, compared with cell proliferation following treatment with gemcitabine or carboplatin alone. In combination, the effects of gemcitabine and carboplatin were synergistic in 3 of 5 cell lines and additive in the other 2. Conclusions and Clinical Relevance: Gemcitabine had antitumor effects on canine TCC cells in vitro, and the combination of gemcitabine and carboplatin had synergistic activity at biologically achievable concentrations.
Afficher plus [+] Moins [-]Evaluation of a commercially available radioimmunoassay and species-specific ELISAs for measurement of high concentrations of insulin in equine serum
2012
Objective: To evaluate a human radioimmunoassay (RIA) and equine and high-range porcine (hrp) species-specific ELISAs for the measurement of high serum insulin concentrations in ponies. Samples: Serum samples from 12 healthy nonobese ponies (7 clinically normal and 5 laminitis prone; 13 to 26 years of age; 11 mares and 1 gelding) before and after glucose, insulin, and dexamethasone administration. Procedures: Intra-and interassay repeatability, freeze-thaw stability, dilutional parallelism, and assay agreement were assessed. Results: Assay detection limits were as follows: RIA, < 389 μU/mL; equine ELISA, < 175 μU/mL; and hrp ELISA, 293 to 8,775 μU/mL. Mean ± SD intra- and interassay repeatability were respectively as follows: RIA, 6.5 ± 5.1 % and 74 ± 3.4%; equine ELISA, 10.6 ± 11.0% and 9.0 ± 4.6%; and hrp ELISA, 19.9 ± 172% and 173 ± 16.6%. Freezing and thawing affected measured concentrations. Dilutional parallelism in the RIA was only evident when insulin-depleted equine serum was used as a diluent (percentage recovery, 95.7 ± 274%); in the ELISAs, dilutional parallelism was observed when a zero calibrator was used. Agreement between RIA and equine ELISA results was good for samples containing concentrations < 175 μU of insulin/mL (bias, −18.5 ± 25.5 μU/mL; higher in RIA). At higher concentrations, assay agreement was poor between RIA and equine ELISA results (bias, −185.3 ± 98.7 μU/mL) and between RIA and hrp ELISA results (bias, 25.3 ± 183.0 μU/mL). Conclusions and Clinical Relevance: Agreement among results of the 3 assays was variable, and dilutional parallelism was only evident with the RIA when insulin-depleted equine serum was tested. Caution is recommended when evaluating high insulin concentrations measured with the RIA or ELISAs.
Afficher plus [+] Moins [-]Effects of oral prednisone administration with or without ultralow-dose acetylsalicylic acid on coagulation parameters in healthy dogs
2012
Objective: To determine the effects of oral prednisone administration with or without ultralow-dose acetylsalicylic acid on coagulation parameters in healthy dogs and to assess intraindividual variation in thromboelastography results. Animals: 14 healthy research dogs and 10 healthy client-owned dogs. Procedures: In a randomized controlled trial, research dogs underwent thromboelastography twice (3 days apart), and intraindividual variation in test results was calculated. Dogs were given prednisone (2 mg/kg/d, PO) plus acetylsalicylic acid (0.5 mg/kg/d, PO) or prednisone (2 mg/kg/d, PO) plus a placebo for 14 days, after which thromboelastography and other tests were repeated. Differences from preadministration (baseline) test results between and within groups were compared. In a separate trial, client-owned dogs also underwent thromboelastography twice 2 days apart to assess intraindividual variation in untreated dogs. Results: Intraindividual variation in thromboelastography results for research dogs was ≤ 10% for maximum amplitude (MA) and α angle. In the research dogs, MA and fibrinogen values significantly increased from baseline, whereas percentage lysis 30 minutes after attainment of the MA as well as antithrombin activity significantly decreased within each group. In the dogs that received prednisone plus a placebo, percentage lysis 60 minutes after attainment of the MA was significantly lower than at baseline. For all parameters for research dogs, there was no difference between groups for change from baseline. Intraindividual variation in findings for client-owned dogs was similar to the variation for research dogs. Conclusions and Clinical Relevance: Prednisone administration resulted in hypercoagulability in healthy dogs as indicated by an increase in MA and plasma fibrinogen concentration and a decrease in antithrombin activity. Concurrent ultralow-dose acetylsalicylic acid use had no effect on measured thromboelastography values. The high intraindividual variation in some thromboelastography parameters may preclude routine use of this technique in clinical practice.
Afficher plus [+] Moins [-]Ultrasonography and noncontrast computed tomography of medial retropharyngeal lymph nodes in healthy cats
2012
Objective: To determine various measurements of medial retropharyngeal lymph nodes (MRPLNs) in healthy cats via ultrasonography and CT. Animals: 45 cats (age range, 2 to 8 years). Procedures: Cats underwent CT of the head and ultrasonography of the cervical region. Various measurements of MRPLNs were obtained, and parenchymal heterogeneity, presence of a hilus, appearance of margins, and attenuation of MRPLNs were determined. Results: Data for 7 cats were excluded because they did not meet inclusion criteria; data for 38 cats were evaluated. Measurements of left and right MRPLNs were not significantly different. Mean length × rostral height × rostral width dimensions of MRPLNs were 20.7 × 12.4 × 3.7 mm and 20.7 × 13.1 × 4.7 mm in ultrasonographic and CT images, respectively. Maximum MRPLN dimensions were approximately 32 × 20 × 7 mm. Mean attenuation of MRPLNs was 40.2 Hounsfield units. Parenchyma of MRPLNs was mildly (via CT) to moderately (via ultrasonography) heterogeneous. A hilus was identified in 95% (via ultrasonography) and 24% or 92% (via CT [depending on criteria used to define a hilus]) of MPRLNs. Lymph node margins were smooth in CT images and mildly irregular in ultrasonographic images. A negative linear correlation was detected between age of cat and MRPLN volume. Conclusions and Clinical Relevance: MRPLNs in cats were easily imaged via ultrasonography and CT. Left and right MRPLNs were symmetric, and MRPLNs were larger in young adult cats versus old cats. Data were intended to serve as references for evaluation of MRPLNs in healthy cats.
Afficher plus [+] Moins [-]Viscoelastic pharmacodynamics after dalteparin administration to healthy dogs
2012
Objective: To evaluate the pharmacodynamic effects of dalteparin in dogs by means of viscoelastic coagulation monitoring with a thromboelastograph and a dynamic viscoelastic coagulometer. Animals: 6 healthy adult mixed-breed dogs. Procedures: Dalteparin (175 U/kg, SC, q 12 h) was administered for 4 days (days 1 through 4). Viscoelastic coagulation monitoring was performed hourly on the first and last days of treatment and included intermittent measurement of anti–activated coagulation factor X activity (AXA). Results: Dalteparin administration resulted in progressive hypocoagulability. On both day 1 and 4, activated clotting time and clot rate for the dynamic viscoelastic coagulometer differed significantly from baseline values, whereas the platelet function parameter did not change on day 1 but did on day 4. The R (reaction time), time from reaction time until the amplitude of the thromboelastography tracing is 20 mm, α-angle, and maximum amplitude differed from baseline values on days 1 and 4, although many thromboelastographic variables were not determined. The AXA was increased from baseline values at 3 and 6 hours after administration of the dalteparin injection on days 1 and 4, and all dogs had AXA values between 0.5 and 1.0 U/mL at 2 and 4 hours after administration. The AXA correlated well with activated clotting time (r = 0.761) and with R (r = 0.810), when values were available. Thromboelastography could not be used to distinguish AXA > 0.7 U/mL. Conclusions and Clinical Relevance: Viscoelastic coagulation monitoring with strong coagulation activators may be used to monitor treatment with dalteparin in healthy dogs.
Afficher plus [+] Moins [-]Simplified procedure for the estimation of glomerular filtration rate following intravenous administration of iodixanol in cats
2012
Objective: To compare the use of a single-sample method involving IV administration of iodixanol with a multisample method involving inulin for the estimation of glomerular filtration rate (GFR) in cats. Animals: 24 cats, including 15 healthy cats and 9 cats with naturally occurring renal diseases. Procedures: Each cat was coadministered iodixanol (a nonionic contrast medium; dose providing 40 mg of I/kg) and inulin (50 mg/kg), IV, and blood samples were collected 60, 90, and 120 minutes later. Serum iodixanol and inulin concentrations were determined by means of high-performance liquid chromatography and colorimetry, respectively. Serum urea nitrogen and creatinine concentrations were also measured. Results: Analysis of the data from healthy cats and cats with naturally occurring renal diseases revealed an excellent correlation between GFR values estimated by the multisample and single-sample methods with iodixanol. Likewise, GFR values estimated from the single-sample method with iodixanol were closely correlated with those calculated from the multisample method with inulin. Conclusions and Clinical Relevance: For estimation of GFR in cats, use of a single-sample method with iodixanol, instead of a multisample procedure, may be an expedient tool in both clinical and research settings because of its benefits to patient well-being as a result of reduced stress associated with blood sample collection.
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