OBJECTIVE To validate primer sets for use in reverse transcription quantitative PCR assays to measure gene expression of cytosolic phospholipase A2 (cPLA2) and microsomal prostaglandin E2 synthase 1 (mPGES1) in equine mononuclear cells and determine the effects of firocoxib, a selective cyclooxygenase 2 (COX-2) inhibitor, on COX-2, cPLA2, and mPGES1 gene expression following incubation of mononuclear cells with lipopolysaccharide (LPS). ANIMALS 8 healthy adult horses. PROCEDURES Peripheral blood mononuclear cells were isolated by density gradient centrifugation and incubated at 37°C with medium alone, firocoxib (100 ng/mL), LPS (1 ng/mL or 1 μg/mL), or combinations of firocoxib and both LPS concentrations. After 4 hours, supernatants were collected and tested for prostaglandin E2 (PGE2) concentration with an enzyme inhibition assay, and gene expression in cell lysates was measured with PCR assays. RESULTS Primer pairs for cPLA2 and mPGES1 yielded single products on dissociation curve analyses, with mean assay efficiencies of 102% and 100%, respectively. Incubation with firocoxib and LPS significantly decreased PGE2 supernatant concentrations and significantly reduced COX-2 and mPGES1 gene expression, compared with values following incubation with LPS alone. CONCLUSIONS AND CLINICAL RELEVANCE Primer sets for mPGES1 and cPLA2 gene expression in equine mononuclear cells were successfully validated. Firocoxib significantly decreased LPS-induced COX-2 and mPGES1 expression, suggesting that it may be useful in the control of diseases in which expression of these genes is upregulated.

OBJECTIVE To establish a simplified single-blood-sample method (SBSM) involving iodixanol to estimate glomerular filtration rate (GFR) in dogs and compare data provided by that procedure with data provided by a conventional multiple-blood-sample method (MBSM) involving inulin. ANIMALS 26 healthy dogs and 36 dogs with naturally occurring renal disease. PROCEDURES Dogs were used in various preliminary experiments to establish protocols for the SBSM and the MBSM of GFR estimation. To evaluate the relationship between GFRs obtained by the SBSM and the MBSM each involving iodixanol, iodixanol (40 mg of I/kg) was administered IV to 26 healthy dogs and 36 dogs with renal disease; blood sample collection was performed before and at 60, 90, and 120 minutes after the injection. To evaluate the relationship between GFRs obtained by the SBSM involving iodixanol and the MBSM involving inulin, iodixanol (40 mg of I/kg) and inulin (50 mg/kg) were coadministered IV to 22 healthy dogs and 3 dogs with renal disease, followed by blood sample collection 30, 60, 90, and 120 minutes later. Serum iodixanol and inulin concentrations were separately determined by reverse-phase high-performance liquid chromatography. RESULTS Findings revealed a correlation (r = 0.99) between GFR estimated by the SBSM and MBSM each involving iodixanol. Likewise, GFR estimated by the SBSM involving iodixanol was correlated (r = 0.89) with that estimated by the MBSM involving inulin. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that the SBSM involving iodixanol can be applied to estimate GFR in dogs, instead of use of an MBSM.

Porcine diarrhea outbreaks caused by porcine epidemic diarrhea virus (PEDV) has occurred in China with significant losses of piglets since 2010. In this study, the complete S and ORF3 genes of 15 field PEDV isolates in mid-eastern China from 2011 to 2013 were detected and compared with other reference strains. Based on S gene, all of the PEDV strains could be assigned to 3 genogroups. Only 1 isolate, JS120103, belonged to genogroup 1 and showed a close relationship with previous Chinese strains DX and JS-2004-2, European strain CV777, and Korean strain DR13. The other 14 isolates belonged to genogroup 3 and showed a close relationship with other Chinese strains isolated after 2010. The S genes of those isolates were 9 nucleotides longer in length than JS120103 and the other reference strains in genogroup 1, with 15 bp insertion and 6 bp deletion. Homology analyses revealed that all of the Chinese field isolates, except JS120103, are 97.6% to 100% (95.8% to 100%) identical in nucleotide (deduced amino acid) sequence to each other. Meanwhile, based on the ORF3 gene, all of the PEDV isolates could be separated into 3 genogroups. Eleven of the 15 field isolates in this study belonged to genogroup 3 and were 95.8% to 100% identical in nucleotide sequence or 95.6% to 100% in deduced amino acid sequence to each other. Our results indicate that the variant PEDV strain spread wildly in mid-eastern China. This will be useful to take into consideration in the control and prevention of this disease.

OBJECTIVE To investigate values of spectral indices for use in predicting responses in dogs during determination of minimum alveolar concentration (MAC) of sevoflurane. ANIMALS 15 healthy German Shepherd Dogs. PROCEDURES Sevoflurane MAC was determined by use of tail clamping. Entropy indices consisting of response entropy and state entropy were recorded during MAC determination. Optimal cutoff points of response entropy and state entropy for use in predicting responses to tail clamping were analyzed with multiple logistic regression. RESULTS Sevoflurane MAC ranged from 1.8% to 2.6% (mean ± SD, 2.2 ± 0.3%). Response entropy and state entropy were significantly higher during positive responses to tail clamping (88 ± 2 and 76 ± 2, respectively) than during negative responses to tail clamping (63 ± 3 and 52 ± 3, respectively). The difference between the 2 entropy indices did not differ between positive (11 ± 1) and negative (13 ± 1) responses to tail clamping. Response entropy and state entropy served as independent predictors of a positive response, with areas under the curve for receiver operating characteristic curves 0.810 (95% confidence interval, 0.716 to 0.903) and 0.828 (95% confidence interval, 0.741 to 0.916), respectively. Optimal cutoff points to predict a positive response were 75 for response entropy and 65 for state entropy, which corresponded to mean ± SD ORs of 25.2 ± 15.6 and 14.9 ± 7.9, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Response entropy and state entropy were good predictors of responses to tail clamping elicited during determination of sevoflurane MAC in healthy dogs.

OBJECTIVE To determine effects of increasing plasma fentanyl concentrations on the minimum alveolar concentration (MAC) of isoflurane in rabbits. ANIMALS 6 adult female New Zealand White rabbits (Oryctolagus cuniculus). PROCEDURES Rabbits were anesthetized with isoflurane in oxygen; ventilation was controlled and body temperature maintained between 38.5° and 39.5°C. Fentanyl was administered IV by use of a computer-controlled infusion system to achieve 6 target plasma concentrations. Isoflurane MAC was determined in duplicate by use of the bracketing technique with a supramaximal electrical stimulus. Blood samples were collected for measurement of plasma fentanyl concentration at each MAC determination. The MAC values were analyzed with a repeated-measures ANOVA followed by Holm-Sidak pairwise comparisons. RESULTS Mean ± SD plasma fentanyl concentrations were 0 ± 0 ng/mL (baseline), 1.2 ± 0.1 ng/mL, 2.2 ± 0.3 ng/mL, 4.4 ± 0.4 ng/mL, 9.2 ± 0.4 ng/mL, 17.5 ± 2.6 ng/mL, and 36.8 ± 2.4 ng/mL. Corresponding mean values for isoflurane MAC were 1.92 ± 0.16%, 1.80 ± 0.16%, 1.60 ± 0.23%, 1.46 ± 0.22%, 1.12 ± 0.19%, 0.89 ± 0.14%, and 0.70 ± 0.15%, respectively. Isoflurane MAC for plasma fentanyl concentrations ≥ 2.2 ng/mL differed significantly from the baseline value. In 3 rabbits, excessive spontaneous movement prevented MAC determination at the highest plasma fentanyl concentration. CONCLUSIONS AND CLINICAL RELEVANCE Fentanyl reduced isoflurane MAC by approximately 60% in New Zealand White rabbits. Further studies will be needed to investigate the cardiorespiratory effects of isoflurane and fentanyl combinations in rabbits; however, fentanyl may prove to be a useful adjunct to inhalation anesthesia in this species.

OBJECTIVE To isolate and characterize endothelial colony-forming cells (ECFCs; a subtype of endothelial progenitor cells) from peripheral blood samples of horses. SAMPLE Jugular venous blood samples from 24 adult horses. PROCEDURES Blood samples were cultured in endothelial cell growth medium. Isolated ECFCs were characterized by use of functional assays of fluorescence-labeled acetylated low-density lipoprotein (DiI-Ac-LDL) uptake and vascular tubule formation in vitro. Expression of endothelial (CD34, CD105, vascular endothelial growth factor receptor 2, and von Willebrand factor) and hematopoietic (CD14) cell markers was assessed through indirect immunofluorescence assay and flow cytometry. The number of passages before senescence was determined through serial evaluation of DiI-Ac-LDL uptake, vascular tubule formation, and cell doubling rates. RESULTS Samples from 3 horses produced colonies at 12 ± 2.5 days with characteristic endothelial single layer cobblestone morphology and substantial outgrowth on expansion. Equine ECFCs formed vascular tubules in vitro and had uptake of DiI-Ac-LDL (74.9 ± 14.7% positive cells). Tubule formation and DiI-Ac-LDL uptake diminished by passage 5. Equine ECFCs tested positive for von Willebrand factor, vascular endothelial growth factor receptor 2, CD34, and CD105 with an immunofluorescence assay and for CD14 and CD105 via flow cytometry. CONCLUSIONS AND CLINICAL RELEVANCE ECFCs can be isolated from peripheral blood of horses and have characteristics similar to those described for other species. These cells may have potential therapeutic use in equine diseases associated with ischemia or delayed vascularization.

OBJECTIVE To determine the intraocular pressure (IOP) in healthy dromedary camels (Camelus dromedarius). ANIMALS 24 clinically normal dromedary camels. PROCEDURES For each camel, the IOP of both eyes was measured with applanation tonometry. Three measurements with < 5% variance were obtained for each eye on the same day of the week for 3 consecutive weeks. Mean IOP was calculated for each eye on each day for comparison purposes. RESULTS Mean ± SD IOPs for the right (31.1 ± 2.1 mm Hg) and left (30.8 ± 1.9 mm Hg) eyes of immature camels were significantly higher than those for the right (27.1 ± 1.2 mm Hg) and left (28.2 ± 1.2 mm Hg) eyes of mature camels. Intra-assay and interassay coefficients of variation (CVs) for IOP measurements of the right and left eyes did not differ significantly between immature and mature camels. Interassay CVs of IOP measurements for the right and left eyes ranged from 1.5% to 12.1% and 1.2% to 10.3%, respectively, for immature camels and from 1.2% to 17.2% and 1.7% to 18.8%, respectively, for mature camels. Intra-assay CVs of IOP measurements for the right and left eyes ranged from 1.5% to 10.6% and 1.9% to 9.6%, respectively, for immature camels and from 2.8% to 16.9% and 2.7% to 12.4%, respectively, for mature camels. Age was negatively correlated (r = −0.403) with IOP. CONCLUSIONS AND CLINICAL RELEVANCE Results provided a reference and might aid in the diagnosis of glaucoma and uveitis during complete ophthalmic examinations of dromedary camels.

Mannheimia haemolytica and Histophilus somni are frequently isolated from diseased cattle with bovine respiratory disease (BRD). They compromise animal lung function and the immune responses generated are not sufficient to limit infection. Identification of specific immunogenic antigens for vaccine development represents a great challenge. Immunogenic proteins were identified by immunoproteomic approach with sera from cattle immunized with a commercial cellular vaccine of M. haemolytica and H. somni. Proteins of M. haemolytica were identified as solute ABC transporter, iron-binding protein, and hypothetical protein of capsular biosynthesis. Histophilus somni proteins correspond to porin, amino acid ABC transporter, hypothetical outer membrane protein, cysteine synthase, and outer membrane protein P6. Although these antigens share strong similarities with other proteins from animal pathogens, the ABC system proteins have been associated with virulence and these proteins could be considered as potential vaccine candidates for BRD.

OBJECTIVE To evaluate pharmacokinetics, recovery times, and recovery quality in horses anesthetized with 1.2 times the minimum alveolar concentration of sevoflurane or desflurane. ANIMALS 6 healthy adult horses. PROCEDURES Anesthesia was maintained with sevoflurane or desflurane for 2 hours at 1.2 times the minimum alveolar concentration. Horses recovered without assistance. During recovery, end-tidal gas samples were collected until horses spontaneously moved. Anesthetic concentrations were measured by use of gas chromatography. After a 1-week washout period, horses were anesthetized with the other inhalation agent. Video recordings of anesthetic recovery were evaluated for recovery quality on the basis of a visual analogue scale by investigators who were unaware of the anesthetic administered. Anesthetic washout curves were fit to a 2-compartment kinetic model with multivariate nonlinear regression. Normally distributed interval data were analyzed by means of paired Student t tests; ordinal or nonnormally distributed data were analyzed by means of Wilcoxon signed rank tests. RESULTS Horses recovered from both anesthetics without major injuries. Results for subjective recovery evaluations did not differ between anesthetics. Area under the elimination curve was significantly smaller and time to standing recovery was significantly less for desflurane than for sevoflurane, although distribution and elimination constants did not differ significantly between anesthetics. CONCLUSIONS AND CLINICAL RELEVANCE Differences in area under elimination the curve between anesthetics indicated more rapid clearance for desflurane than for sevoflurane in horses, as predicted by anesthetic blood solubility differences in this species. More rapid elimination kinetics was associated with faster recovery times, but no association with improved subjective recovery quality was detected.

OBJECTIVE To measure incidence and estimate temporal and spatial dynamics of porcine reproductive and respiratory syndrome virus (PRRSV) infection in US sow herds. ANIMALS 371 sow herds in the United States from 14 production companies. PROCEDURES The exponentially weighted moving average was used to monitor incident PRRSV infections for onset of an epidemic. The spatial scan statistic was used to identify areas at significantly high risk of PRRS epidemics. A χ2 test was used to estimate whether there were significant differences in the quarterly and annual PRRS incidence among time periods, and a bivariable logistic regression model was used to estimate whether PRRSV infection during a given year increased the odds of that herd being infected in the following year. RESULTS During the 4-year period of this study, 29% (91/319; 2009 to 2010), 33% (106/325; 2010 to 2011), 38% (135/355; 2011 to 2012), and 32% (117/371; 2012 to 2013) of the herds reported new infections. Weekly incidence was low during spring and summer and high during fall and winter. The exponentially weighted moving average signaled the onset of a PRRSV epidemic during the middle 2 weeks of October each year. Disease incidence was spatially clustered. Infection in the previous year increased the odds of infection in 2010 to 2011 and 2011 to 2012. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated a striking repeatability in annual PRRSV temporal and spatial patterns across 4 years of data among herds from 14 production companies, which suggested that efforts to control PRRSV at a regional level should continue to be supported.