OBJECTIVE To identify the genetic cause for congenital photosensitivity and hyperbilirubinemia (CPH) in Southdown sheep. ANIMALS 73 Southdown sheep from a CPH research flock and 48 sheep of various breeds from commercial flocks without CPH. PROCEDURES Whole-genome sequencing was performed for a phenotypically normal Southdown sheep heterozygous for CPH. Heterozygous variants within Slco1b3 coding exons were identified, and exons that contained candidate mutations were amplified by PCR assay methods for Sanger sequencing. Blood samples from the other 72 Southdown sheep of the CPH research flock were used to determine plasma direct and indirect bilirubin concentrations. Southdown sheep with a plasma total bilirubin concentration < 0.3 mg/dL were classified as controls, and those with a total bilirubin concentration ≥ 0.3 mg/dL and signs of photosensitivity were classified as mutants. Sanger sequencing was used to determine the Slco1b3 genotype for all sheep. Genotypes were compared between mutants and controls of the CPH research flock and among all sheep. Protein homology was measured across 8 species to detect evolutionary conservation of Slco1b. RESULTS A nonsynonymous mutation at ovine Chr3:193,691,195, which generated a glycine-to-arginine amino acid change within the predicted Slco1b3 protein, was significantly associated with hyperbilirubinemia and predicted to be deleterious. That amino acid was conserved across 7 other mammalian species. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested a nonsynonymous mutation in Slco1b3 causes CPH in Southdown sheep. This disease appears to be similar to Rotor syndrome in humans. Sheep with CPH might be useful animals for Rotor syndrome research.

The objective of this study was to observe the outcomes of adding an antimicrobial treatment to a conventional treatment regime in horses with severe equine asthma in a clinical setting. Eleven client-owned horses with a history consistent with severe equine asthma, increased respiratory effort and nostril flaring, ≥ 20% neutrophils on bronchoalveolar lavage (BAL), and a positive tracheal wash (TW) bacterial culture were treated with environmental management, corticosteroids, and bronchodilators. Six horses were also treated with an antimicrobial (principal group), while the other 5 were administered saline as a placebo (control group). Treatment with antimicrobials significantly improved the post-treatment clinical score of the principal group compared with the pre-treatment score, whereas no significant difference occurred in the control group. The principal group also had significantly less neutrophil myeloperoxidase (MPO) activity post-treatment than pre-treatment, with a median difference of -0.39 units/[protein] in the principal group and a median difference of -0.21 units/[protein] in the controls. There was no difference in MPO activity pre- versus post-treatment in the control group. No differences were noted in the intra-group comparisons of pre- versus post-treatment BAL neutrophil counts, mucus scores, and concentrations of interleukin-8 (IL-8) or tumor necrosis factor-alpha (TNF-α) in bronchoalveolar lavage fluid (BALF) in either group. There were no differences found in the inter-group comparisons of the principal versus controls for each of the pre- and post-treatment time periods for BAL neutrophil count, mucus score, clinical scores, MPO activity, and IL-8 or TNF-α concentrations. The role of airway bacteria in horses with severe equine asthma requires further investigation as antimicrobial therapy improved post-treatment clinical scores and decreased MPO activity in the group of horses studied, but did not affect other measures of airway inflammation.

OBJECTIVE To compare time to achieve vascular access (TTVA) between an ultrasound-guided technique (UST) and landmark-based technique (LMT) for central venous catheter (CVC) placement in healthy anesthetized dogs. ANIMALS 39 purpose-bred hounds. PROCEDURES Anesthetized dogs that were hemodynamically stable following completion of a terminal surgical exercise were enrolled in the study during 2 phases, with a 45-day intermission between phases. For each dog, a UST and LMT were used for CVC placement via each external jugular vein by 2 operators (criticalist and resident). The TTVA and number of venipuncture attempts and catheter redirections were recorded for each catheterization. Placement of the CVC was confirmed by contrast fluoroscopy. After euthanasia, a gross dissection was performed during which a hematoma score was assigned to the catheter insertion site. For each phase, nonlinear least squares estimation was used for learning curve analysis of the UST. RESULTS Median TTVA, number of venipuncture attempts and catheter redirections, and hematoma score did not differ significantly between the 2 operators for either technique. Median TTVA for the UST (45 seconds) was significantly longer than that for the LMT (7 seconds). Learning curve analysis indicated that 8 and 7 UST catheterizations were required to achieve performance stability in phases 1 and 2, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that the UST was comparable to the LMT for CVC placement in healthy dogs. The extra time required to perform the UST was not clinically relevant. Additional studies evaluating the UST for CVC placement in clinically ill dogs are warranted.

The H9N2 strains of avian influenza viruses (AIVs) circulate worldwide in poultry and cause sporadic infection in humans. To better understand the evolution of these viruses while circulating in poultry, an H9N2 chicken isolate was passaged 19 times in chickens via aerosol inoculation. Whole-genome sequencing showed that the viruses from the initial stock and those after the 8th and 19th passages (P0, P8, and P19) all had the same monobasic cleavage site in the hemagglutinin (HA), typical for viruses of low pathogenicity. However, at position 226 of the HA protein the ratio of glutamine (which favors avian-type receptor binding) to leucine (which favors mammalian-type receptor binding) decreased from 54:46 in P0, to 87:13 in P8, and then 0:100 in P19. In chickens exposed to aerosols of P0, P8, or P19, replication of the viruses was similar and mainly limited to the respiratory tract. None of the infected chickens showed any clinical signs. Over the 19 passages the viruses maintained relatively stable infectivity but gradually lost lethality to chicken embryos. According to the hemagglutination inactivation assay, P8 was slightly and P19 significantly (P < 0.05) less thermostable than P0. Collectively, after 19 passages in chickens the H9N2 AIVs retained low pathogenicity with a positive selection of L226 in the HA. These findings suggest that H9N2 viruses might acquire mammalian specificity after asymptomatic circulation in avian species.

The purpose of this study was to determine a wide range of selected hematologic, venous blood gases, and plasma biochemistry analytes in common chameleons (Chamaeleo chamaeleon). Blood samples were collected from the ventral tail vein of 41 common chameleons to determine reference intervals for 30 different blood analytes. The calcium-to-phosphorus ratio, packed cell volume (PCV), refractometric total solids (TS), blood cell counts, and differentials were also determined. The microscopic evaluation of blood smears revealed inclusion bodies in monocytes in 7 of the samples. Females showed significantly higher values of plasma proteins and calcium and cholesterol concentrations and males showed significantly higher values of aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) plasma concentrations. Significant differences were found between similar analytes determined by different testing methodologies in the PCV/hematocrit, electrolytes (sodium, potassium), and plasma proteins [TS, total protein (TP) and albumin]. Blood analytes determined in this study can provide baseline data that may be useful when evaluating the health status of common chameleons, taking into consideration the potential effects of gender and the type of analyzer used.

OBJECTIVE To determine the in vitro effect of calcitriol on indicators of immune system function in endotoxin-primed blood samples from healthy dogs. SAMPLE Blood samples from 6 healthy adult dogs. PROCEDURES Leukocytes were primed by incubation of blood samples with lipopolysaccharide (LPS; endotoxin) or PBS solution (unprimed control group) for 1 hour. Following priming, blood samples were incubated with calcitriol (2 × 10(−7)M) or ethanol (control substance) for 24 hours. After sample incubation, LPS-stimulated leukocyte production of tumor necrosis factor (TNF) and interleukin-10 (IL10) was measured with a canine-specific multiplex assay, and apoptosis and toll-like receptor 4 (TLR4) expression were evaluated via flow cytometry. RESULTS LPS stimulation of unprimed leukocytes but not endotoxin-primed leukocytes resulted in a significant increase in TNF and IL10 production, confirming the presence of endotoxin tolerance in dogs in vitro. Endotoxin priming significantly increased neutrophil viability with no effect on lymphocyte viability or TLR4 expression by neutrophils and monocytes. Calcitriol exposure significantly decreased LPS-stimulated production of TNF by unprimed and endotoxin-primed leukocytes. Conversely, calcitriol exposure had no effect on IL10 production by unprimed leukocytes but did significantly increase IL10 production by endotoxin-primed leukocytes. Calcitriol had no significant effect on the degree of neutrophil or lymphocyte apoptosis, nor was neutrophil and monocyte TLR4 expression affected in unprimed or endotoxin-primed leukocytes. CONCLUSIONS AND CLINICAL RELEVANCE These data indicated that calcitriol induced an anti-inflammatory shift in unprimed and endotoxin-primed canine leukocytes in vitro, without compromising neutrophil and monocyte TLR4 expression or altering the viability of neutrophils and lymphocytes in canine blood samples.

OBJECTIVE To characterize the distribution and intensity of cyclooxygenase (COX)-2 expression in the eyes of cats with and without uveitis and to determine whether COX-2 expression is correlated with severity of inflammation. SAMPLES Archived ocular tissue specimens from 51 cats with and 10 cats without ocular disease. PROCEDURES Specimens from only 1 eye were evaluated for each cat. Specimens were stained with H&E stain or immunohistochemical stain for detection of COX-2 and reviewed. For each eye, the type, severity, and distribution of inflammation and the distribution and intensity of COX-2 expression were determined for the uvea and other ocular tissues. Correlation between COX-2 expression and inflammation severity was also assessed. RESULTS COX-2 was not expressed in any nondiseased eye. Of the 51 diseased eyes, 20 had histologic evidence of lymphocytic-plasmacytic uveitis, 13 had neutrophilic uveitis, 11 had diffuse iris melanoma with uveitis, and 7 had diffuse iris melanoma without uveitis. Of the 44 eyes with uveitis, COX-2 was detected in the uvea of 16, including 11 eyes with lymphocytic-plasmacytic uveitis, 4 with neutrophilic uveitis, and 1 with diffuse iris melanoma–induced uveitis. Inflammation was severe, moderate, or mild in 10, 5, and 1 of those eyes, respectively. Cyclooxygenase-2 was detected in the cornea of 21 eyes with uveitis and 1 eye with diffuse iris melanoma without uveitis. Uveitis severity was positively correlated with COX-2 expression in both the uvea and cornea. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that COX-2 is an inflammatory mediator in feline uveitis but not diffuse iris melanoma.

OBJECTIVE To assess the in vitro effects of doxorubicin and tetrathiomolybdate (TM) on cells from a canine hemangiosarcoma cell line. SAMPLE Cultured cells from the canine hemangiosarcoma–derived cell line DEN-HSA. PROCEDURES Cells were treated with TM (0 to 1.5μM), doxorubicin (0 to 5μM), or both with or without 24 hours of pretreatment with ascorbic acid (750μM). Degree of cellular cytotoxicity was measured with a colorimetric assay. Long-term growth inhibition was assessed with a 10-day colony-formation assay. Induction of apoptosis was quantitated by fluorometric assessment of caspase-3 and −7 activation. Formation of reactive oxygen species (ROS) was also detected fluorometrically. RESULTS Exposure of cells to the combination of TM and doxorubicin resulted in a greater decrease in proliferation and clonogenic survival rates than exposure to each drug alone. This treatment combination increased ROS formation and apoptosis to a greater extent than did doxorubicin or TM alone. Ascorbic acid inhibited both TM-induced ROS formation and apoptosis. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that the enhancement in cytotoxic effects observed with DEN-HSA cell exposure to the combination of doxorubicin and TM was achieved through an increase in ROS production. These findings provide a rationale for a clinical trial of this treatment combination in dogs with hemangiosarcoma.

OBJECTIVE To assess effects of speed on kinematic variables measured by use of extremity-mounted inertial measurement units (IMUs) in nonlame horses performing controlled exercise on a treadmill. ANIMALS 10 nonlame horses. PROCEDURES 6 IMUs were attached at predetermined locations on 10 nonlame Franches Montagnes horses. Data were collected in triplicate during trotting at 3.33 and 3.88 m/s on a high-speed treadmill. Thirty-three selected kinematic variables were analyzed. Repeated-measures ANOVA was used to assess the effect of speed. RESULTS Significant differences between the 2 speeds were detected for most temporal (11/14) and spatial (12/19) variables. The observed spatial and temporal changes would translate into a gait for the higher speed characterized by increased stride length, protraction and retraction, flexion and extension, mediolateral movement of the tibia, and symmetry, but with similar temporal variables and a reduction in stride duration. However, even though the tibia coronal range of motion was significantly different between speeds, the high degree of variability raised concerns about whether these changes were clinically relevant. For some variables, the lower trotting speed apparently was associated with more variability than was the higher trotting speed. CONCLUSIONS AND CLINICAL RELEVANCE At a higher trotting speed, horses moved in the same manner (eg, the temporal events investigated occurred at the same relative time within the stride). However, from a spatial perspective, horses moved with greater action of the segments evaluated. The detected changes in kinematic variables indicated that trotting speed should be controlled or kept constant during gait evaluation.

OBJECTIVE To compare bronchoalveolar lavage (BAL) accomplished by use of a bronchoscopic (B-BAL) and a nonbronchoscopic (NB-BAL) technique in healthy cats. ANIMALS 12 healthy cats. PROCEDURES Two BALs were performed in a randomized order 2 weeks apart in each cat. Cats were anesthetized, and a 2.9-mm fiberoptic bronchoscope (B-BAL) or 8F red rubber catheter (NB-BAL) was wedged in a bronchus. Two 5-mL aliquots of saline (0.9% NaCl) solution were infused into the left and right caudal lung fields and aspirated manually with a 20-mL syringe. Proportion of BAL fluid (BALF) retrieved, depth of wedging, and anesthetic complications were recorded. Total nucleated cell count, differential cell count, and semiquantitative scores of cytologic slide quality were determined for all BALF samples. Results were compared with ANOVAs and Wilcoxon signed rank tests. RESULTS Proportion of retrieved BALF and depth of wedging were significantly greater for B-BAL than NB-BAL. Differential cell counts and cytologic slide quality did not differ significantly between techniques. Complications included transient hemoglobin desaturation (24/24 [100%] BALs) and prolonged anesthetic recovery time (4/24 [17%] BALs). Anesthetic recovery scores did not differ significantly between techniques. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that NB-BAL was noninferior to B-BAL with regard to ease of performance, anesthetic variables, and cytologic slide quality for cats without clinical respiratory tract disease.