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Evaluation of a broad range real-time polymerase chain reaction (RT-PCR) assay for the diagnosis of septic synovitis in horses Полный текст
2013
Elmas, Colette R. | Koenig, Judith B. | Bienzle, Dorothee | Cribb, Nicola C. | Cernicchiaro, Natalia | Coté, Nathalie M. | Weese, J Scott
Septic synovitis is a potentially debilitating and life-threatening disorder in horses. We hypothesized that a universal bacterial real-time PCR (RT-PCR) assay would have improved sensitivity and decreased turn-around time for detection of bacteria in synovial fluid (SF) samples. Forty-eight SF samples were collected from 36 horses that presented to two referral institutions with suspected septic synovitis. Universal RT-PCR, bacterial culture and SF analysis were performed on all samples, and an interpretation on the sample being septic or not was derived by three board certified specialists from the history, clinical assessment and SF characteristics. RT-PCR results were compared to a composite standard comprised of positive culture and interpretation by all three specialists of samples as “septic”. For 41 of 48 samples (85%), culture and RT-PCR results were concordant. Compared to the composite standard, 83% of samples were correctly classified by RT-PCR (turn-around time of approximately 4 hours). Relative sensitivity and specificity of RT-PCR were 87% and 72% respectively, and 56% and 86% for culture. Hence, universal RT-PCR was a rapid and highly sensitive test, which may accelerate diagnosis and improve outcome for horses with septic synovitis.
Показать больше [+] Меньше [-]Evaluation of the efficacy and safety of single administration of 4.7-mg deslorelin acetate implants on egg production and plasma sex hormones in Japanese quail (Coturnix coturnix japonica) Полный текст
2013
Petritz, Olivia A. | Sanchez-Migallon Guzman, David | Paul-Murphy, Joanne | Fecteau, Kellie | Mete, Asli | Kass, Philip H. | Hawkins, Michelle G.
Objective: To evaluate the effects of 4.7-mg deslorelin acetate implants on egg production and plasma concentrations of 17β-estradiol and androstenedione in Japanese quail (Coturnix coturnix japonica) over 180 days and assess safety of the implants in quail via gross and histologic examination. Animals: 20 female Japanese quail. Procedures: Following a 7-day period of consistent egg laying, quail were anesthetized and received a 4.7-mg deslorelin implant (treatment group; n = 10) or identical placebo implant (control group; 10) SC between the scapulae. Egg production was monitored daily. Plasma concentrations of 17β-estradiol and androstenedione were measured on days 0 (immediately prior to implant injection), 14, 29, 62, 90, 120, 150, and 180 via radioimmunoassay. Birds were weighed periodically and euthanized at day 180 for complete necropsy. Results: Egg production was significantly decreased in the treatment group, compared with the control group, from 2 to 12 weeks after implant injection. Egg production ceased in 6 of 10 quail in the treatment group (mean duration of cessation, 70 days). Plasma androstenedione and 17β-estradiol concentrations were significantly lower on day 29 in the treatment group than in the control group. Plama androstenedione and 17β-estradiol concentrations were significantly lower on day 29 in the treatment group then in the control group. Conclusions and Clinical Relevance: 4.7-mg deslorelin acetate implants reversibly decreased egg laying for approximately 70 days in most of the Japanese quail evaluated. Further studies evaluating implants containing different concentrations of the drug are needed in quail and other avian species.
Показать больше [+] Меньше [-]Gene biomarkers in peripheral white blood cells of horses with experimentally induced osteoarthritis Полный текст
2013
Kamm, J Lacy | Frisbie, David D. | McIlwraith, C Wayne | Orr, Kindra E.
Objective: To use microarray analysis to identify genes that are differentially expressed in horses with experimentally induced osteoarthritis. Animals: 24 horses. Procedures: During arthroscopic surgery, a fragment was created in the distal aspect of the radiocarpal bone in 1 forelimb of each horse to induce osteoarthritis. At day 14 after osteoarthritis induction, horses began exercise on a treadmill. Blood and synovial fluid samples were collected before and after surgery. At day 70, horses were euthanized and tissues were harvested for RNA analysis. An equine-specific microarray was used to measure RNA expression in peripheral WBCs. These data were compared with mRNA expression (determined via PCR assay) in WBCs, cartilage, and synovium as well as 2 protein biomarkers of cartilage matrix turnover in serum and synovial fluid. Results: A metalloproteinase domain-like protein decysin-1 (ADAMDEC1), glucose-regulated protein (GRP) 94, hematopoietic cell signal transducer (HCST), Unc-93 homolog A (hUNC-93A), and ribonucleotide reductase M2 polypeptide (RRM2) were significantly differentially regulated in WBCs of horses with osteoarthritis, compared with values prior to induction of osteoarthritis. There was correlation between the gene expression profile in WBCs, cartilage, and synovium and the cartilage turnover proteins. Gene expression of ADAMDEC1, hUNC-93A, and RRM2 in WBCs were correlated when measured via microarray analysis and PCR assay. Conclusions and Clinical Relevance: Expression of ADAMDEC1, GRP94, HCST, hUNC-93A, and RRM2 was differentially regulated in peripheral WBCs obtained from horses with experimentally induced osteoarthritis. Gene expression of ADAMDEC1, hUNC-93A, and RRM2 in peripheral WBCs has the potential for use as a diagnostic aid for osteoarthritis in horses.
Показать больше [+] Меньше [-]Evaluation of tissue Doppler imaging for regional quantification of radial left ventricular wall motion in healthy horses Полный текст
2013
Decloedt, Annelies | Verheyen, Tinne | Sys, Stanislas | Clercq, Dominique de | Loon, Gunther van
Objective: To compare the feasibility and repeatability of tissue Doppler imaging (TDI) for quantification of radial left ventricular (LV) velocity and deformation from different imaging planes and to correlate cardiac event timing data obtained by TDI to M-mode and pulsed-wave Doppler-derived time intervals in horses. Animals: 10 healthy adult horses. Procedures: Repeated echocardiography was performed by 2 observers from right and left parasternal short-axis views at papillary muscle and chordal levels. The TDI measurements of systolic and diastolic velocity, strain rate, strain peak values, and timing were performed in 8 LV wall segments (LV free wall and interventricular septum from right parasternal views; left and right region of LV wall from left parasternal views). The inter- and intraobserver within- and between-day variability and measurement variability were assessed. The correlation between TDI-based measurements and M-mode and pulsed-wave Doppler-based time measurements was calculated. Results: TDI measurements of velocity, strain rate, and strain were feasible in each horse, although deformation could often not be measured in the LV free wall. Systolic and diastolic time intervals could be determined with low to moderate variability, whereas peak amplitude variability ranged from low to high. The TDI-based time measurements were significantly correlated to M-mode and pulsed-wave Doppler measurements. Conclusions and Clinical Relevance: TDI measurements of radial LV velocity and deformation were feasible with low to moderate variability in 8 LV segments. These measurements can be used for evaluating LV function in further clinical studies.
Показать больше [+] Меньше [-]Effect of hydroxyethyl starch 130/0.4 and 200/0.5 solutions on canine platelet function in vitro Полный текст
2013
McBride, Duana | Hosgood, Giselle L. | Mansfield, Caroline S. | Smart, Lisa
Objective-To determine whether dilution of blood samples from healthy dogs with 2 hydroxyethyl starch (HES) solutions, HES 130/0.4 and HES 200/0.5, would result in platelet dysfunction as measured by closure time (Ct) beyond a dilutional effect. Sample-Citrated blood samples from 10 healthy dogs with a Ct within reference limits (52 to 86 seconds). Procedures-Blood samples were diluted 1:9 and 1:3 with 6% HES 130/0.4 and 10% HES 200/0.5 solutions and saline (0.9% NaCl) solution. Dilutions at 1:9 and 1:3 mimicked 10 mL/kg and 30 mL/kg doses, respectively, ignoring in vivo redistribution. Closure time was measured with a platelet function analyzer and compared among dilutions. Results-A dilutional effect on Ct was evident for the 1:3 dilution, compared with the 1:9 dilution, but only HES 200/0.5 increased the Ct beyond the dilutional effect at the 1:3 dilution, to a median Ct of 125 seconds (interquartile range, 117.5 to 139.5 seconds). No effect of HES or dilution on Ct was identified at the 1:9 dilution. Conclusions and Clinical Relevance-1:3 dilution of blood samples from healthy dogs with HES 200/0.5 but not HES 130/0.4 significantly increased Ct beyond the dilutional effect, suggesting that IV administration of HES 200/0.5 in dogs might cause platelet dysfunction.
Показать больше [+] Меньше [-]Evaluation of neutrophil apoptosis in horses with acute abdominal disease Полный текст
2013
Krista, Kathryn M. | White, N. A. (Nathaniel A) | Barrett, Jennifer G. | Furr, Martin O. | Buechner-Maxwell, Virginia A.
Objective-To quantify peripheral blood neutrophil apoptosis in equine patients with acute abdominal disease (ie, colic) caused by strangulating or nonstrangulating intestinal lesions and compare these values with values for horses undergoing elective arthroscopic surgery. Animals-20 client-owned adult horses. Procedures-Peripheral blood was collected from horses immediately prior to and 24 hours after surgery for treatment of colic (n = 10) or elective arthroscopic surgery (10), and neutrophils were counted. Following isolation by means of a bilayer colloidal silica particle gradient and culture for 24 hours, the proportion of neutrophils in apoptosis was detected by flow cytometric evaluation of cells stained with annexin V and 7-aminoactinomycin D. Values were compared between the colic and arthroscopy groups; among horses with colic, values were further compared between horses with and without strangulating intestinal lesions. Results-Percentage recovery of neutrophils was significantly smaller in preoperative samples (median, 32.5%) and in all samples combined (35.5%) for the colic group, compared with the arthroscopy group (median, 66.5% and 58.0%, respectively). No significant differences in the percentages of apoptotic neutrophils were detected between these groups. Among horses with colic, those with strangulating intestinal lesions had a significantly lower proportion of circulating apoptotic neutrophils in postoperative samples (median, 18.0%) than did those with nonstrangulating lesions (66.3%). Conclusions and Clinical Relevance-The smaller proportion of apoptotic neutrophils in horses with intestinal strangulation suggested that the inflammatory response could be greater or prolonged, compared with that of horses with nonstrangulating intestinal lesions. Further investigations are needed to better understand the relationship between neutrophil apoptosis and inflammation during intestinal injury.
Показать больше [+] Меньше [-]Comparison of overground and treadmill-based gaits of dogs Полный текст
2013
Torres, Bryan T. | Moëns, Noel M.M. | Al-Nadaf, Sami | Reynolds, Lisa R. | Fu, Yang-Chieh | Budsberg, Steven C.
Objective-To compare overground and treadmill-based gaits of dogs. Animals -5 clinically normal adult mixed-breed dogs. Procedures-To obtain dynamic gait data, 30 retroreflective markers were affixed bilaterally to specific regions of the hind limbs and pelvis of each dog. For each dog, 3-D joint motion data (sagittal [flexion and extension], transverse [internal and external rotation], and frontal [abduction and adduction] planes of motion) for the hip, femorotibial, and tarsal joints were acquired during walking and trotting through a calibrated testing space overground or on a treadmill. Comparison of data was performed via generalized indicator function analysis and Fourier analysis. Results-Both overground and treadmill-based gaits produced similar waveforms in all planes of motion. Fourier analysis revealed no difference between overground and treadmill-based gaits in the sagittal plane of motion; however, small differences were detected between overground and treadmill-based gaits in the other 2 planes of motion. Additionally, femorotibial joint motion during walking did not differ among planes of motion. Generalized indicator function analysis was able to detect differences between overground and treadmill-based gait waveforms in all planes of motion for all joints during walking and trotting. Conclusions and Clinical Relevance-In dogs, overground and treadmill-based gaits produced similar waveform shapes. Of the 3 planes of motion evaluated, only sagittal plane kinematic gait data were unaffected by mode of ambulation as determined via Fourier analysis. Sagittal kinematic gait data collected from dogs during overground or treadmill-based ambulation were comparable. However, analysis methods may affect data comparisons.
Показать больше [+] Меньше [-]Pharmacokinetics of meloxicam in rabbits after oral administration of single and multiple doses Полный текст
2013
Fredholm, Daniel V. | Carpenter, James W. | KuKanich, Butch | Kohles, Micah
Objective-To determine the pharmacokinetics of meloxicam (1 mg/kg) in rabbits after oral administration of single and multiple doses. Animals-6 healthy rabbits. Procedures-A single dose of meloxicam (1 mg/kg, PO) was administered to the rabbits. After a 10-day washout period, meloxicam (1 mg/kg, PO) was administered to rabbits every 24 hours for 5 days. Blood samples were obtained from rabbits at predetermined intervals during both treatment periods. Plasma meloxicam concentrations were determined, and noncompartmental pharmacokinetic analysis was performed. Results-The mean peak plasma concentration and area under the plasma concentration-versus-time curve extrapolated to infinity after administration of a single dose of meloxicam were 0.83 μg/mL and 10.37 h•μg/mL, respectively. After administration of meloxicam for 5 days, the mean peak plasma concentration was 1.33 μg/mL, and the area under the plasma concentration-versus-time curve from the time of administration of the last dose to 24 hours after that time was 18.79 h•μg/mL. For single- and multiple-dose meloxicam experiments, the mean time to maximum plasma concentration was 6.5 and 5.8 hours and the mean terminal half-life was 6.1 and 6.7 hours, respectively. Conclusions and Clinical Relevance-Plasma concentrations of meloxicam for rabbits in the present study were proportionally higher than those previously reported for rabbits receiving 0.2 mg of meloxicam/kg and were similar to those determined for animals of other species that received clinically effective doses. A dose of 1 mg/kg may be necessary to achieve clinically effective circulating concentrations of meloxicam in rabbits, although further studies are needed.
Показать больше [+] Меньше [-]Changes in leukocyte populations of cows with milk fever or displaced abomasum after calving Полный текст
2013
Ohtsuka, Hiromichi | Fukuda, Shigeo | Kudo, Katsunori | Tomioka, Michiko | Koiwa, Masateru | Kimura, Kayoko
Most of the metabolic diseases of dairy cows occur within the first 2 wk after calving, and cows with a metabolic disease are prone to infectious diseases. Although metabolic diseases are generally recognized as a risk factor for infectious diseases owing to the associated decrease in immune function, the difference in immune status between cows with milk fever (MF) or displaced abomasum (DA) during the lactation period has not been clarified. Therefore, the peripheral blood leukocyte populations in 38 multiparous Holstein cows from 1 herd were analyzed after calving. The cows were divided into 3 groups according to health: 21 cows that remained clinically healthy throughout the experimental period (control group), 9 cows that had MF on the day of calving, and 8 cows with an onset of DA within 4 wk after calving. The T- and B-cell numbers were lowest at week 0, and they increased gradually after calving. There was no significant difference between the 3 groups in the number of each subset of leukocytes on the day of calving, but the number of CD8+ T-cells was significantly lower in the MF and DA groups than in the control group at week 1. The numbers of CD4+, CD8+, and WC1+ T-cells tended to be lower in the DA group than in control group from weeks 4 to 12, a tendency not observed in the MF group. These data suggest that when cows have DA around the time of calving, their lymphocyte numbers remain lower until 12 wk after calving.
Показать больше [+] Меньше [-]Comparison of histological lesions in mink with acute hemorrhagic pneumonia associated with Pseudomonas aeruginosa or Escherichia coli Полный текст
2013
Salomonsen, Charlotte Mark | Boye, Mette | Høiby, Niels | Jensen, Trine H. | Hammer, Anne Sofie
Hemorrhagic pneumonia can be a major cause of mortality in farmed mink in the fall. In its classic form, hemorrhagic pneumonia is caused by the bacterium Pseudomonas aeruginosa. In recent years, however, outbreaks of this type of pneumonia that are associated with hemolytic Escherichia coli have also occurred in farmed mink. The purpose of this study was to compare histological lesions of acute hemorrhagic pneumonia associated with both P. aeruginosa and E. coli in mink, including a description of tissue distribution of pathogens, in an attempt to differentiate between the 2 disease entities based on histopathology. The study included material submitted for diagnostic investigation to the National Veterinary Institute in Denmark from 2006 to 2009. Altogether, 19 cases of hemorrhagic pneumonia with a pure lung culture of P. aeruginosa and 18 cases of hemorrhagic pneumonia with a pure lung culture of E. coli were examined. Formalin-fixed paraffin-embedded lung tissue obtained from the mink was examined by histology and fluorescence in-situ hybridization (FISH). It was possible to detect a slight histological difference between hemorrhagic pneumonia caused by P. aeruginosa and by E. coli, as P. aeruginosa was most often found surrounding blood vessels and lining the alveoli, while E. coli showed a more diffuse distribution in the lung tissue. Furthermore, P. aeruginosa often elicited a very hemorrhagic response in the lung, while infection with E. coli was associated with a higher frequency of alveolar edema and mild lymphoid cuffing in the lungs.
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