Objective: To determine whether a novel optimized plasmid carrying the porcine growth hormone–releasing hormone (GHRH) wild-type cDNA administered at a lower dose was as effective at eliciting physiologic responses as a commercial GHRH plasmid approved for use in Australia. Animals: 134 gilts. Procedures: Estrus was synchronized and gilts were bred. Pregnant gilts were assigned to 2 treatment groups (40 gilts/group) or 1 untreated control group (24 gilts). Gilts in one of the treatment groups received the commercial GHRH plasmid, whereas gilts in the other treatment group received a novel optimized GHRH plasmid; both plasmids were administered IM in the right hind limb, which was followed by electroporation. Sow and litter performance were monitored for the 3 gestations after treatment. Results: A significant increase in insulin-like growth factor-I concentrations, decrease in perinatal mortality rate, increase in the number of pigs born alive, and increase in the weight and number of pigs weaned were detected for both groups receiving the GHRH-expressing plasmids, compared with values for the control group. Additionally, there was a significant decrease in sow attrition in GHRH-treated females, compared with attrition in the control group, during the 3 gestations after treatment. Conclusions and Clinical Relevance: Both of the GHRH plasmids provided significant benefits for sow performance and baby pig survivability for pregnant and lactating sows and their offspring during the 3 gestations after treatment, compared with results for untreated control gilts. Use of a novel optimized plasmid reduced the effective plasmid dose in these large mammals.

Objective: To determine the extent to which a hydroxyapatite coating promotes pin stability in the third metacarpal bone during transfixation casting in horses. Animals: 14 adult horses. Procedures: 7 horses each were assigned to either an uncoated or hydroxyapatite-coated pin group. Three transcortical pins were placed in the third metacarpal bone of each horse and incorporated into a cast for 8 weeks. Insertion and extraction torque were measured, and torque reduction was calculated. Radiography was performed at 0, 4, and 8 weeks. Lameness evaluation was performed at 2, 4, 6, and 8 weeks. Bacteriologic culture of pins and pin holes was performed at pin removal. Results: All horses used casts without major complication throughout the study. Insertion torque was higher in uncoated pins. There was no effect of group on extraction torque. Hydroxyapatite-coated pins had lower torque reduction. Five of 15 hydroxyapatite-coated pins maintained or increased stability, whereas all uncoated pins loosened. Pin hole radiolucency, lameness grades, and positive bacteriologic culture rates were not different between groups. Conclusions and Clinical Relevance: Hydroxyapatite coating increased pin stability within the third metacarpal bone of horses during 8 weeks of transfixation casting but did not improve pin performance on clinical assessments. Clinical use of hydroxyapatite-coated transfixation pins may result in greater pin stability; however, further research is necessary to improve the consistency of pin osteointegration and elucidate whether clinical benefits will ultimately result from this approach in horses.

Objective: To determine whether preanalytic and analytic factors affect evaluation of the urinary protein-to-creatinine (UPC) ratio in dogs. Sample: 50 canine urine samples. Procedures: The UPC ratio was measured to assess the intra-assay imprecision (20 measurements within a single session), the influence of predilution (1:10, 1:20, and 1:100) for urine creatinine concentration measurement, and the effect of storage at room temperature (approx 20°C), 4°C, and −20°C. Results: The coefficient of variation at room temperature determined with the 1:20 predilution was < 10.0%, with the highest coefficients of variation found in samples with a low protein concentration or low urine specific gravity. This variability could result in misclassification of samples with UPC ratios close to the thresholds defined by the International Renal Interest Society to classify dogs as nonproteinuric (0.2), borderline proteinuric (0.21 to 0.50), or proteinuric (> 0.51). A proportional bias was found in samples prediluted 1:10, compared with samples prediluted 1:20 or 1:100. At room temperature, the UPC ratio did not significantly increase after 2 and 4 hours. After 12 hours at room temperature and at 4°C, the UPC ratio significantly increased. The UPC ratio did not significantly change during 3 months of storage at −20°C. Conclusions and Clinical Relevance: The intra-assay precision of the UPC ratio was sufficiently low to avoid misclassification of samples, except for values close to 0.2 or 0.5. The optimal predilution ratio for urine creatinine concentration measurement was 1:20. A 1:100 predilution is recommended in samples with a urine specific gravity > 1.030. The UPC ratio must be measured as soon as samples are collected. Alternatively, samples should be immediately frozen to increase their stability and minimize the risk of misclassification of proteinuria.

Objective-To measure the effects of tidal volume, ventilatory frequency, and oxygen insufflation flow on the fraction of inspired oxygen in cadaveric horse heads attached to a lung model. Sample-8 heads of equine cadavers. Procedures-Each cadaveric horse head was intubated with a nasotracheal tube that extended into the proximal portion of the trachea. Oxygen was delivered through an oxygen catheter contained within and extending to the tip of the nasotracheal tube. The trachea was connected to the lung model by use of a spiral-wound hose with a sampling adaptor. Eight treatment combinations involving 2 tidal volumes (5 and 8 L), 2 ventilatory frequencies (6 and 12 mechanical breathes/min), and 2 insufflation rates (10 and 15 L/min) were applied to each head. Hand-drawn inspired gas samples were collected and analyzed for oxygen concentrations. Results-The fraction of inspired oxygen (measured at mid trachea) ranged from 26.8% to 39.4%. Fraction of inspired oxygen was significantly higher with a smaller tidal volume, lower ventilatory frequency, and higher insufflation rate. Conclusions and Clinical Relevance-In the study model, measured fraction of inspired oxygen varied with ventilatory pattern as well as oxygen insufflation rate. Clinically, this information could be beneficial for interpretation of data regarding arterial blood gases and hemoglobin saturation and in making appropriate oxygen insufflation decisions for anesthetized horses that are breathing room air.

Objective: To determine reference ranges for serum cobalamin (Cbl), urine methylmalonic acid (uMMA), and plasma total homocysteine (tHcys) concentrations and to compare values for healthy control dogs with values for Border Collies (BCs), a breed in which hereditary cobalamin deficiency has been identified. Animals: 113 BCs, 35 healthy control dogs fed a typical diet, and 12 healthy dogs fed a bone and raw food diet exclusively. Procedures: Urine and blood samples were obtained from each dog and Cbl, uMMA, and tHcys concentrations were determined. Results: Reference ranges for Cbl (261 to 1,001 ng/L), uMMA (0 to 4.2 mmol/mol of creatinine), and tHcys (4.3 to 18.4 μmol/L) concentrations were determined. Four BCs had a Cbl concentration lower than the assay detection limit (150 ng/L); median uMMA and tHcys concentrations in these dogs were 4,064 mmol/mol of creatinine and 51.5 μmol/L, respectively. Clinical abnormalities included stunted growth, lethargy, anemia, and proteinuria. Abnormalities improved after administration of cobalamin. Of the 109 healthy BCs with Cbl and tHcys concentrations within reference ranges, 41 (37.6%) had a high uMMA concentration (range, 5 to 360 mmol/mol). Results for dogs fed raw food were similar to those for control dogs. Conclusions and Clinical Relevance: Hereditary cobalamin deficiency is a rare disease with various clinical signs. The finding of methylmalonic aciduria in healthy eucobalaminemic BCs and BCs with clinical signs of Cbl deficiency was surprising and indicated these dogs may have defects in intracellular processing of Cbl or intestinal Cbl malabsorption, respectively. Studies investigating Cbl absorption and metabolic pathways are warranted.

Objective: To determine pharmacokinetics after IV and oral administration of a single dose of tramadol hydrochloride to Hispaniolan Amazon parrots (Amazona ventralis). Animals: 9 healthy adult Hispaniolan Amazon parrots (3 males, 5 females, and 1 of unknown sex). Procedures: Tramadol (5 mg/kg, IV) was administered to the parrots. Blood samples were collected from −5 to 720 minutes after administration. After a 3-week washout period, tramadol (10 and 30 mg/kg) was orally administered to parrots. Blood samples were collected from −5 to 1,440 minutes after administration. Three formulations of oral suspension (crushed tablets in a commercially available suspension agent, crushed tablets in sterile water, and chemical-grade powder in sterile water) were evaluated. Plasma concentrations of tramadol and its major metabolites were measured via high-performance liquid chromatography. Results: Mean plasma tramadol concentrations were > 100 ng/mL for approximately 2 to 4 hours after IV administration of tramadol. Plasma concentrations after oral administration of tramadol at a dose of 10 mg/kg were < 40 ng/mL for the entire time period, but oral administration at a dose of 30 mg/kg resulted in mean plasma concentrations > 100 ng/mL for approximately 6 hours after administration. Oral administration of the suspension consisting of the chemical-grade powder resulted in higher plasma tramadol concentrations than concentrations obtained after oral administration of the other 2 formulations; however, concentrations differed significantly only at 120 and 240 minutes after administration. Conclusions and Clinical Relevance: Oral administration of tramadol at a dose of 30 mg/kg resulted in plasma concentrations (> 100 ng/mL) that have been associated with analgesia in Hispaniolan Amazon parrots.

Objective: To compare the axial stiffness, maximum axial displacement, and ring deformation during axial loading of single complete and incomplete circular (ring) external skeletal fixator constructs. Sample: 32 groups of single ring constructs (5 constructs/group). Procedures: Single ring constructs assembled with 2 divergent 1.6-mm-diameter Kirschner wires were used to stabilize a 60-mm-long segment of 16-mm-diameter acetyl resin rod. Construct variables included ring type (complete or incomplete), ring diameter (50, 66, 84, or 118 mm), and fixation wire tension (0, 30, 60, or 90 kg). Axial loading was performed with a materials testing system. Construct secant stiffness and maximum displacement were calculated from the load-displacement curves generated for each construct. Ring deformation was calculated by comparing ring diameter during and after construct loading to ring diameter prior to testing. Results: Complete ring constructs had greater axial stiffness than did the 66-, 84-, and 118-mm-diameter incomplete ring constructs. As fixation wire tension increased, construct stiffness increased in the 66-, 84-, and 118-mm-diameter incomplete ring constructs. Maximum axial displacement decreased with increasing fixation wire tension, and complete ring constructs allowed less displacement than did incomplete ring constructs. Incomplete rings were deformed by wire tensioning and construct loading. Conclusions and Clinical Relevance: Mechanical performance of the 66-, 84-, and 118-mm-diameter incomplete ring constructs improved when wire tension was applied, but these constructs were not as stiff as and allowed greater displacement than did complete ring constructs of comparable diameter. For clinical practice, tensioning the wires placed on 84- and 118-mm-diameter incomplete rings to 60 kg is recommended.

Objective: To evaluate and compare bone modeling and remodeling in fractured and non-fractured central tarsal bones (CTBs) of racing Greyhounds. Sample: Paired cadaveric tarsi from 6 euthanized racing Greyhounds with right CTB fractures and 6 racing Greyhounds with other nontarsal injuries. Procedures: CTBs were dissected and fractured CTBs were reconstructed. Central tarsal bones were evaluated through standard and nonscreen high-detail radiography, computed tomography, and histologic examination. The bone mineral density (BMD) was calculated adjacent to fracture planes and as a gradient on sagittal computed tomographic images. Sagittal and transverse plane sections of bone were obtained and submitted for subjective histologic assessment. Linear mixed-effects models were used to compare findings. Results: Fractured right CTBs had greater BMD in the dorsal and midbody regions of the sagittal plane sections than did nonfractured CTBs. The BMD ratios from bone adjacent to the dorsal slab fracture planes were not different between fractured and nonfractured right CTBs. Conclusions and Clinical Relevance: Findings supported the existence of site-specific bone adaptation in CTBs of Greyhounds, with modeling and remodeling patterns that were unique to fractured right CTBs. The dorsal and midbody regions of fractured bones had greater BMD, and fractures occurred through these zones of increased BMD.

Objective: To determine the toxicokinetics of N-(methylsuccinimido)anthranoyllycoctonine–type low larkspur alkaloids in beef cattle. Animals: 5 Black Angus steers and 35 Swiss Webster mice. Procedures: Low larkspur (Delphinium andersonii) was collected, dried, ground, and administered to 5 steers via oral gavage to provide a dose of 12 mg of N-(methylsuccinimido)-anthranoyllycoctonine alkaloids/kg. Steers were housed in metabolism crates for 96 hours following larkspur administration; heart rate was monitored continuously, and blood samples were collected periodically for analysis of serum concentrations of 16-deacetylgeyerline, methyllycaconitine, geyerline, and nudicauline and assessment of kinetic parameters. The LD50 of a total alkaloid extract from D andersonii was determined in Swiss Webster mice. Results: The alkaloids were quickly absorbed, with a maximum serum concentration achieved within 18 hours after administration. Geyerline and nudicauline coeluted as 1 peak and were considered together for toxicokinetic analysis. Mean ± SD elimination half-life was 18.4 ± 4.4 hours, 15.6 ± 1.5 hours, and 16.5 ± 5.1 hours for 16-deacetylgeyerline, methyllycaconitine, and geyerline and nudicauline, respectively. There were significant differences in maximum serum concentration, amount absorbed, and distribution half-life among the 4 alkaloids. The mouse LD50 was 9.8 mg/kg. Conclusions and Clinical Relevance: Results suggested that clinical poisoning was likely to be most severe approximately 18 hours after exposure. Cattle should be closely monitored for at least 36 hours after initial exposure. Additionally, a withdrawal time of approximately 7 days would be required to clear > 99% of the toxic alkaloids from the serum of cattle that have ingested low larkspur.

Africa has the highest burden of infectious diseases in the world and yet the least capacity for its risk management. It has therefore become increasingly important to search for ‘fit-for- purpose’ approaches to infectious disease surveillance and thereby targeted disease control. The fact that the majority of human infectious diseases are originally of animal origin means we have to consider One Health (OH) approaches which require inter-sectoral collaboration for custom-made infectious disease surveillance in the endemic settings of Africa. A baseline survey was conducted to assess the current status and performance of human and animal health surveillance systems and subsequently a strategy towards OH surveillance system was developed. The strategy focused on assessing the combination of participatory epidemiological approaches and the deployment of mobile technologies to enhance the effectiveness of disease alerts and surveillance at the point of occurrence, which often lies in remote areas. We selected three study sites, namely the Ngorongoro, Kagera River basin and Zambezi River basin ecosystems. We have piloted and introduced the next-generation Android mobile phones running the EpiCollect application developed by Imperial College to aid geo-spatial and clinical data capture and transmission of this data from the field to the remote Information Technology (IT) servers at the research hubs for storage, analysis, feedback and reporting. We expect that the combination of participatory epidemiology and technology will significantly improve OH disease surveillance in southern Africa.