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Use of serum amyloid A in serum and synovial fluid to detect eradication of infection in experimental septic arthritis in horses
2020
Yoshimura, Seiji | Koziy, Roman V. | Dickinson, Ryan | Moshynskyy, Igor | McKenzie, Joscelyn A. | Simko, Elemir | Bracamonte, Jose L.
While serum amyloid A (SAA) has been investigated as a potential marker for septic arthritis in horses, no study has reported on whether SAA can be used to detect eradication of joint infection. Therefore, the objective of this study was to investigate whether the eradication of joint infection in experimentally induced septic arthritis in horses can be detected using serum and synovial fluid SAA. A total of 17 horses were randomly assigned to 3 groups. A middle carpal joint of each horse was injected with saline (control group, n = 3), lipopolysaccharide (LPS) (nonseptic synovitis group, n = 6), or Escherichia coli (septic arthritis group, n = 8) on day 0. Starting on day 1, horses underwent treatment for septic arthritis. Sequential samples of serum and synovial fluid were collected, and quantification of SAA was carried out. Concentrations of serum and synovial fluid SAA were compared among groups and time points. A concurrent study was conducted and determined that infection was eradicated on day 4 in this experimental model of septic arthritis. Concentrations of serum and synovial fluid SAA rapidly increased after inoculation of E. coli and were highest on day 3 and day 4, respectively. Thereafter, both serum and synovial fluid SAA decreased with eradication of joint infection, although they remained significantly increased from baseline until day 9 and day 10, respectively. Serum and synovial fluid SAA did not increase in the control or nonseptic synovitis group. These findings suggest that serial measurements rather than a single measurement of SAA are required to determine eradication of infection from septic arthritis in horses.
Show more [+] Less [-]Development and assessment of a novel ex vivo corneal culture technique involving an agarose-based dome scaffold for use as a model of in vivo corneal wound healing in dogs and rabbits
2020
Berkowski, William M. Jr | Gibson, Daniel J. | Craft, Serena L. | Whitley, Robert D. | Schultz, Gregory S. | Plummer, Caryn E.
OBJECTIVE To develop and assess a novel ex vivo corneal culture technique involving an agarose-based dome scaffold (ABDS) for use as a model of in vivo corneal wound healing in dogs and rabbits. SAMPLE Corneas from clinically normal dogs (paired corneas from 8 dogs and 8 single corneas) and rabbits (21 single corneas). PROCEDURES 8 single dog corneas (DCs), 1 DC from each pair, and 10 rabbit corneas (RCs) were wounded with an excimer laser; 1 DC from each pair and 11 RCs remained unwounded. Corneas were cultured for 21 days on ABDSs (8 pairs of DCs and all RCs) or on flat-topped scaffolds (8 single DCs). The surface area of corneal fluorescein retention was measured every 6 (DCs) or 12 (RCs) hours until full corneal epithelialization was detected. Changes in corneal clarity were evaluated at 0, 7, 14, and 21 days. RESULTS Median time to full epithelialization for wounded dog and rabbit corneas was 48 and 60 hours, respectively; among wounded DCs, time to full epithelization did not differ by scaffold type. After 21 days of culture on ABDSs, all DCs and RCs that epithelialized developed a circular, diffuse, cloud-like pattern of optical haze, whereas DCs cultured on flat-topped scaffolds developed a focal, crater-like region of optical haze. All corneas on the ABDSs maintained convex curvature throughout the study. CONCLUSIONS AND CLINICAL RELEVANCE Wounded ex vivo DCs and RCs cultured on ABDSs reliably epithelialized, formed optical haze (consistent with in vivo wound healing), and maintained convex curvature. This culture technique may be adaptable to other species.
Show more [+] Less [-]Correlation of Activity Data in Normal Dogs to Distance Traveled
2020
Eskander, Bishoy S. | Barbar, Megan | Evans, Richard B. | Enomoto, Mastaka | Lascelles, B. Duncan X. | Conzemius, Michael G.
The objective of this study was to explore the mathematical relationships between independent variables (patient morphometrics and treadmill speed) and dependent variables (accelerometer or pedometer output) when evaluating data from accelerometers and pedometers in dogs. Twenty dogs took part in 3 randomized activities, consisting of exercise on a treadmill at 1.0, 1.5, and 2.0 m/s for a total distance of 1 km at each speed. Dogs simultaneously wore both an accelerometer and a pedometer. Statistical analysis used multiple regression models to discover the relationships between independent and dependent variables. A formula was developed to predict the distance traveled by a dog based on its morphometrics and activity monitor output. Shoulder height had stronger correlations to accelerometer and pedometer outputs than other morphometric variables. As shoulder height increased, all accelerometer and pedometer outputs decreased. As treadmill speed increased, both accelerometer and pedometer step counts decreased, while accelerometer activity counts increased. According to a formula derived to predict the total distance traveled using patient shoulder height and accelerometer or pedometer output, pedometer steps were the most accurate predictor of distance traveled. Accelerometer steps were less accurate when using the same model. Accelerometer activity counts did not reveal a meaningful predictive formula. The results of this study indicate that patient morphometrics and treadmill speed (as a measure of intensity) influenced accelerometer and pedometer data. The pedometer data more precisely and accurately estimated the distance traveled based on step counts and patient shoulder height. In normal dogs, accelerometer and pedometer steps may reasonably estimate distance traveled.
Show more [+] Less [-]In vitro holding security of four friction knots of monofilament or multifilament suture used as a first throw for vascular ligation
2020
Tremolada, Giovanni | Hazenfield, Kurt M. | Smeak, Daniel D.
OBJECTIVE To evaluate holding security of 4 friction knots created with various monofilament and multifilament sutures in a vascular ligation model. SAMPLE 280 friction knot constructs. PROCEDURES 10 friction knots of 4 types (surgeon's throw, Miller knot, Ashley modification of the Miller knot, and strangle knot) created with 2-0 monofilament (polyglyconate, polydioxanone, poliglecaprone-25, and glycomer-631) and braided multifilament (silk, lactomer, and polyglactin-910) sutures were separately tied on a mock pedicle and pressure tested to the point of leakage. Linear regression analysis was performed to compare leakage pressures among suture materials (within friction knot type) and among knot types (within suture material). RESULTS Mean leakage pressure of surgeon's throws was significantly lower than that of all other knots tested, regardless of the suture material used. All the other knots had mean leakage pressures considered supraphysiological. Significant differences in mean leakage pressure were detected between various friction knots tied with the same type of suture and various suture types used to create a given knot. Variability in leakage pressure among knots other than the surgeon's throw was greatest for poliglecaprone-25 and lowest for polydioxanone. CONCLUSIONS AND CLINICAL RELEVANCE Most differences in knot security, although statistically significant, may not have been clinically relevant. However, results of these in vitro tests suggested the surgeon's throw should be avoided as a first throw for pedicle ligation and that poliglecaprone-25 may be more prone to friction knot slippage than the other suture materials evaluated.
Show more [+] Less [-]Novel use of an activity monitor to model jumping behaviors in cats
2020
Sharon, Kate P. | Thompson, Caryn M. | Lascelles, B. Duncan X. | Parrish, Rudolph S.
OBJECTIVE To develop methods to identify and characterize activity monitor (AM) data signatures for jumps performed by cats. ANIMALS 13 healthy, client-owned cats without evidence of osteoarthritis or degenerative joint disease. PROCEDURES Each cat was fitted with the same AM, individually placed in an observation room, then simultaneously recorded by 3 video cameras during the observation period (5 to 8 hours). Each cat was encouraged to jump up (JU), jump down (JD), and jump across (JA) during the observation period. Output from the AM was manually annotated for jumping events, each of which was characterized by functional data analysis yielding relevant coefficients. The coefficients were then used in linear discriminant analysis to differentiate recorded jumps as JUs, JDs, or JAs. To assess the model's ability to distinguish among the 3 jump types, a leave-one-out cross-validation method was used, and the misclassification error rate of the overall categorization of the model was calculated. RESULTS Of 731 jumping events, 29 were misclassified. Overall, the mean misclassification error rate per cat was 5.4% (range, 0% to 12.5%), conversely indicating a correct classification rate per cat of 94.6%. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that the model was successful in correctly identifying JUs, JDs, and JAs in healthy cats. With advancements in AM technology and data processing, there is potential for the model to be applied in clinical settings as a means to obtain objective outcome measures.
Show more [+] Less [-]Histologic assessment of ligament vascularity and synovitis in dogs with cranial cruciate ligament disease
2019
Kuroki, Keiichi | Harbin, Robert | Ikeda, Hitoshi | Bozynski, Chantelle C. | Leary, Emily | Cook, James L.
OBJECTIVE: To assess the relationship between histologic degeneration of cranial cruciate ligaments (CCLs) and severity of synovitis and ligament vascularity. SAMPLE: CCL and synovium from 59 stifle joints (53 dogs). PROCEDURES: CCL and synovium specimens were obtained from stifle joints of juvenile (15 joints; 12 dogs) and adult (25 joints; 22 dogs) dogs with intact CCLs and dogs with CCL rupture (rCCL; 19 joints; 19 dogs). Vascular density and degenerative changes of the CCL core region and severity of synovitis were semiquantitatively evaluated. Relationships were analyzed by use of a random effects model to account for correlated specimens. RESULTS: Mean ± SD modified Bonar scores (scale, 0 to 9) of adults (4.85 ± 0.44) and dogs with rCCL (5.69 ± 0.49) were significantly higher than scores of juveniles (1.13 ± 0.55). Vascularity scores (scale, 0 to 3) were significantly higher for juveniles (3.00 ± 0.24) than for adults (1.53 ± 0.27) and dogs with rCCL (0.78 ± 0.23). Synovitis scores were not significantly different among groups. There was a significant negative relationship between modified Bonar scores and vascularity scores for juveniles and adults and for adults and dogs with rCCL when controlling for age, but there was not a significant relationship between modified Bonar scores and synovitis scores. There was a significant relationship between modified Bonar scores and body weight of adults. CONCLUSIONS AND CLINICAL RELEVANCE: Poor blood supply to the core region could be an important underlying condition for spontaneous degeneration of the CCL in at-risk dogs.
Show more [+] Less [-]Use of standard diagnostic techniques to determine eradication of infection in experimental equine septic arthritis
2019
Koziv, R. V. | Yoshimura, S. | Dickinson, R. | Rybicka, J. M. | Moshynskyy, I. | Ngeleka, M. | Bracamonte, J. L. | Simko, E.
Septic arthritis is an important disease in horses, necessitating aggressive and prolonged therapy. In order to guide therapy, reliable methods of detecting the eradication of infection are needed. Therefore, the objective of this study was to investigate detection of eradication of infection in an experimental model of equine septic arthritis using standard diagnostic techniques. For this purpose, 17 adult horses were assigned to 3 experimental groups. The middle carpal joint of each horse was injected with Escherichia coli (Septic group, n = 8), lipopolysaccharide (LPS) (LPS group, n = 6), or sterile saline (Control group, n = 3) at day 0. Contralateral joints were not injected. Standard therapy was applied to all joints except non-injected joints in the Control group at day 1. Sequential samples of synovial fluid (SF) were collected for bacterial culture using 3 culture media [Columbia blood agar (CBA), brain heart infusion broth (BHI), and Signal blood culture medium] and for cytological evaluation [percentage neutrophils (PN), total nucleated cell count (TNCC), and total protein (TP)]. Escherichia coli-specific polymerase chain reaction (PCR) was carried out to detect E. coli DNA in synovial fluid. Culture and PCR were positive for E. coli in all joints injected with E. coli at day 1 and 1 joint was positive on BHI at day 4. Based on the results of bacterial culture, PCR, and TNCC, the elimination of infection in our experimental model occurred by day 4 post-infection in 6 out of 7 cases. Total protein (TP) and PN remained elevated at clinical threshold used for diagnosis of septic arthritis until day 14. In our experimental model of E. coli-induced arthritis, we conclude that TP and PN may not be good indicators for detecting the eradication of bacterial infection caused by E. coli from infected and subsequently treated joints.
Show more [+] Less [-]Comparison of 3 vaccination strategies against porcine reproductive and respiratory syndrome virus, Mycoplasma hyopneumoniae, and porcine circovirus type 2 on a 3 pathogen challenge model
2018
Jeong, J. | Kang, I. | Kim, S. | Park, K. H. | Park, C. | Chae, C.
The objective of this study was to compare clinical, microbiologic, immunologic, and pathologic parameters in pigs each concurrently administered porcine reproductive and respiratory syndrome virus (PRRSV), Mycoplasma hyopneumoniae, and porcine circovirus type 2 (PCV2) vaccine from 1 of 2 commercial sources at 21 days of age and challenged with field strains of each of the 3 pathogens. Pigs were challenged with PRRSV and M. hyopneumoniae at 42 days of age (-14 days post-challenge, dpc) followed by a challenge with PCV2 at 56 days of age (0 dpc). Significant differences were observed between vaccinated challenged and unvaccinated challenged groups in clinical (average daily gain and clinical signs), microbiologic (viremia and nasal shedding), immunologic (antibodies and interferon-γ secreting cells), and pathologic (lesions) outcomes. Significant differences were observed among the 3 vaccinated challenged groups in microbiologic (nasal shedding of M. hyopneumoniae and viremia of PCV2) and immunologic (M. hyopneumoniae- and PCV2-specific interferon-γ secreting cells) outcomes. The vaccination regimen for PRRSV vaccine, M. hyopneumoniae vaccine, and PCV2 vaccine is efficacious for controlling triple challenge with PRRSV, M. hyopneumoniae, and PCV2 from weaning to finishing period.
Show more [+] Less [-]Pharmacokinetics of tranexamic acid in healthy dogs and assessment of its antifibrinolytic properties in canine blood
2018
Osekavage, Katie E. | Brainard, Benjamin M. | Lane, Selena L. | Almoslem, Mohammed | Arnold, Robert D. | Koenig, Amie
OBJECTIVE To assess pharmacokinetics of tranexamic acid (TXA) in dogs and assess antifibrinolytic properties of TXA in canine blood by use of a thromboelastography-based in vitro model of hyperfibrinolysis. ANIMALS 6 healthy adult dogs. PROCEDURES Dogs received each of 4 TXA treatments (10 mg/kg, IV; 20 mg/kg, IV; approx 15 mg/kg, PO; and approx 20 mg/kg, PO) in a randomized crossover-design study. Blood samples were collected at baseline (time 0; immediately prior to drug administration) and predetermined time points afterward for pharmacokinetic analysis and pharmacodynamic (thromboelastography) analysis by use of an in vitro hyperfibrinolysis model. RESULTS Maximum amplitude (MA [representing maximum clot strength]) significantly increased from baseline at all time points for all treatments. The MA was lower at 360 minutes for the 10-mg/kg IV treatment than for other treatments. Percentage of clot lysis 30 minutes after MA was detected was significantly decreased from baseline at all time points for all treatments; at 360 minutes, this value was higher for the 10-mg/kg IV treatment than for other treatments and higher for the 20-mg/kg IV treatment than for the 20-mg/kg PO treatment. Maximum plasma TXA concentrations were dose dependent. At 20 mg/kg, IV, plasma TXA concentrations briefly exceeded concentrations suggested for complete inhibition of fibrinolysis. Oral drug administration resulted in a later peak antifibrinolytic effect than did IV administration. CONCLUSIONS AND CLINICAL RELEVANCE Administration of TXA improved clot strength and decreased fibrinolysis in blood samples from healthy dogs in an in vitro hyperfibrinolysis model. Further research is needed to determine clinical effects of TXA in dogs with hyperfibrinolysis.
Show more [+] Less [-]Evaluation of potential predictor variables for PCR assay diagnosis of Anaplasma phagocytophilum infection in equids in Northern California
2018
Langdon Fielding, C. | Rhodes, Diane M. | Howard, Elizabeth J. | Mayer, Jennifer R.
OBJECTIVE To identify clinical or clinicopathologic variables that can be used to predict a positive PCR assay result for Anaplasma phagocytophilum infection in equids. ANIMALS 162 equids. PROCEDURES Medical records were reviewed to identify equids that underwent testing for evidence of A phagocytophilum infection by PCR assay between June 1, 2007, and December 31, 2015. For each equid that tested positive (case equid), 2 time-matched equids that tested negative for the organism (control equids) were identified. Data collected included age, sex, breed, geographic location (residence at the time of testing), physical examination findings, and CBC and plasma biochemical analysis results. Potential predictor variables were analyzed by stepwise logistic regression followed by classification and regression tree analysis. Generalized additive models were used to evaluate identified predictors of a positive test result for A phagocytophilum. RESULTS Total lymphocyte count, plasma total bilirubin concentration, plasma sodium concentration, and geographic latitude were linear predictors of a positive PCR assay result for A phagocytophilum. Plasma creatine kinase activity was a nonlinear predictor of a positive result. CONCLUSIONS AND CLINICAL RELEVANCE Assessment of predictors identified in this study may help veterinarians identify equids that could benefit from early treatment for anaplasmosis while definitive test results are pending. This information may also help to prevent unnecessary administration of oxytetracycline to equids that are unlikely to test positive for the disease.
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