Circulating monocytes and tissue macrophages were suggested to be susceptible to avian reovirus (ARV) infection. To determine if ARV infects and replicates in mononuclear phagocytes (KUL01-positive cells), we infected 3-day-old specific-pathogen-free chickens with ARV strain 2408 by inoculation of the left footpad. The left footpads and spleens were collected for analysis at 1.5 and 2.5 d after inoculation. Replication of ARV in the footpad and spleen was demonstrated by detection of the viral protein σNS using immunohistochemical testing and viral S1 RNA expression by real-time quantitative polymerase chain reaction (qPCR). Furthermore, immunofluorescent double-staining assay of cytocentrifuged cells and cryosections of the footpad and spleen for the viral protein σNS and the surface marker recognized by monoclonal antibody (MAb) KUL01 indicated that KUL01-positive cells costained with MAb H1E1, which recognizes ARV protein σNS. In addition, more ARV S1 RNA was measured by qPCR in the KUL01-positive cell samples prepared from the footpad or spleen 1.5 d after inoculation compared with non-KUL01-positive cell samples. The amounts of ARV S1 RNA in the spleen were significantly lower (P < 0.05) than the amounts in the footpad 1.5 d after inoculation. The results suggest that ARV infects mononuclear phagocytes and then replicates within these cells before migrating to the spleen, where it infects and replicates in KUL01-positive cells.

Atopic dermatitis is a common allergic skin disease in dogs. The aim of this study was to examine the possibility of a correlation between biophysical skin variables: skin hydration (SH), skin pH, and erythema intensity measured in 10 different body regions and both total Canine Atopic Dermatitis Extent and Severity Index (CADESI-03) and CADESI measured in a given region (CADESI L). The study was conducted using 33 dogs with atopic dermatitis. The assessment of the biophysical variables was done in 10 body regions: the lumbar region, right axillary fossa, right inguinal region, ventral abdominal region, right lateral thorax region, internal surface of the auricle, interdigital region of right forelimb, cheek, bridge of nose, and lateral site of antebrachum. Positive correlations were found between SH and CADESI L for the following regions: the inguinal region (r = 0.73) and the interdigital region (r = 0.82), as well as between total CADESI and SH on digital region (r = 0.52). Also, positive correlations were reported for skin pH and CADESI L in the lumbar region (r = 0.57), the right lateral thorax region (r = 0.40), and the lateral antebrachum (r = 0.35). Positive correlations were found in the interdigital region between erythema intensity and the total CADESI-03 (r = 0.60) as well as the CADESI L (r = 0.7). The results obtained suggest that it may be possible to use skin hydration, pH, and erythema intensity to assess the severity of skin lesion but positive correlation was only found in < 13.3% of possible correlations and usage of these measures in dogs is limited.

Multicomponent nutraceuticals are becoming increasingly popular treatments or adjunctive therapies for osteoarthritis in veterinary medicine despite lack of evidence of efficacy for many products. The objective of this study was to evaluate the anti-inflammatory and antioxidant activities of a commercially available C-phycocyanin-based nutraceutical and select constituent ingredients in an in-vitro model of canine osteoarthritis. Normal canine articular chondrocytes were used in an in-vitro model of osteoarthritis. Inflammatory conditions were induced using interleukin-1β. The nutraceutical preparation as a whole, its individual constituents, as well as carprofen were evaluated at concentrations of 0 to 250 μg/mL for reduction of the following inflammatory mediators and indicators of catabolism of the extracellular matrix: prostaglandin E2 (PGE2), tumor necrosis factor-α (TFN-α), interleukin-6 (IL-6), metalloproteinase-3 (MMP-3), nitric oxide, and sulfated glycosaminoglycans (sGAGs). Validated, commercially available assay kits were used for quantitation of inflammatory mediators. The antioxidant capacities, as well as cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), and lipoxygenase (LOX) inhibitory activities of the whole nutraceutical preparation and select constituents, were also assessed using validated commercially available assay kits. The antioxidant capacity of the nutraceutical and constituents was concentration-dependent. The nutraceutical and constituents appear to display anti-inflammatory activity primarily through the inhibition of COX-2. The nutraceutical displayed similar strength to carprofen in reducing TNF-α, IL-6, MMP-3, nitric oxide, and sGAGs at select concentration ranges. The C-phycocyanin (CPC)-based nutraceutical and constituents may be able to mediate 3 primary pathogenic mechanisms of osteoarthritis: inflammation, chondral degeneration, and oxidative stress in vitro. The nutraceutical may be clinically useful in veterinary medicine and its efficacy should be further investigated in vivo.

OBJECTIVE To measure changes in interleukin-8 (IL-8), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) concentrations in stored canine packed RBCs (PRBCs) over time and assess the effect of leukoreduction on these cytokine concentrations. ANIMALS 12 anesthetized healthy Greyhounds. PROCEDURES 1 unit of whole blood from each dog was processed into PRBCs. Half of each PRBCs unit was passed through a leukoreduction filter to produce a leukoreduced unit, and the remaining blood was kept as a nonleukoreduced unit. All units had a CBC performed on day 0 (day of collection) and were stored at 2° to 6°C. Samples were collected from leukoreduced and nonleukoreduced units on days 0, 10, 20, 30, and 37 and centrifuged; the supernatant was stored at −80°C until analysis. Canine TNF-α and IL-8 concentrations were assessed with a multiplexed genomic and proteomic biomarker analyzer, and canine IL-1β concentration was measured by ELISA. RESULTS Leukocyte counts were decreased by ≥ 99.9% in all leukoreduced units. Median TNF-α and IL-1β concentrations were not significantly different between leukoreduced and nonleukoreduced units and did not change significantly during storage; median IL-8 concentration was significantly higher in nonleukoreduced versus leukoreduced units on all days, and was greater at all time points after ≥ 10 days of storage than on day 0. Median IL-8 concentration in leukoreduced units did not increase during storage. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that leukoreduction was effective for the removal of leukocytes from canine PRBCs and prevented significant increases in IL-8 concentration during storage. Further studies are needed to evaluate whether leukoreduction reduces cytokine-associated complications of transfusion.

The aim of this study was to evaluate the antimicrobial activity and determine the minimum bactericidal concentration (MBC) of the essential oils derived from Origanum vulgare (oregano), Melaleuca alternifolia (tea tree), Cinnamomum cassia (cassia), and Thymus vulgaris (white thyme) against Salmonella Typhimurium, Salmonella Enteritidis, Escherichia coli, Staphylococcus aureus and Enterococcus faecalis. The study also investigated the ability of these different bacterial strains to develop adaptation after repetitive exposure to sub-lethal concentrations of these essential oils. The MBC of the essential oils studied was determined by disc diffusion and broth dilution methods. All essential oils showed antimicrobial effect against all bacterial strains. In general, the development of adaptation varied according to the bacterial strain and the essential oil (tea tree > white thyme > oregano). Therefore, it is important to use essential oils at efficient bactericidal doses in animal feed, food, and sanitizers, since bacteria can rapidly develop adaptation when exposed to sub-lethal concentrations of these oils.

During the United States Department of Agriculture (USDA) National Animal Health Monitoring System’s (NAHMS) 2007‐2008 beef study, producers from 24 states were offered the opportunity to evaluate their animals for internal parasites and for overall responses to treatment with anthelmintics. A lapse of 45 d was required between initial sampling and any previous treatments. Choice of anthelmintic (oral benzimidazoles, and both injectable and pour-on endectocides) was at the discretion of the producer so as not to alter the local control programs. Fresh fecal samples were collected from 20 animals, or from the entire group if less than 20, then randomly assigned to 1 of 3 participating laboratories for examination. Analyses consisted of double centrifugation flotation followed by enumeration of strongyle, Nematodirus, and Trichuris eggs (the presence of coccidian oocysts and tapeworm eggs was also noted). Where strongyle eggs per gram (epg) exceeded 30, aliquots from 2 to 6 animals were pooled for egg isolation and polymerase chain reaction (PCR) analysis for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. Results from 72 producers (19 States) indicated that fecal egg count reductions were < 90% in 1/3 of the operations. All operations exhibiting less than a 90% reduction had used pour-on macrocyclic lactones as the anthelmintic treatment. While some of these less than expected reductions could have been the result of improper drug application, PCR analyses of the parasite populations surviving treatment, coupled with follow-up studies at a limited number of sites, indicated that less than expected reductions were most likely due to anthelmintic resistance in Cooperia spp. and possibly Haemonchus spp.

The objective of this study was to develop a murine retinal/choroidal/scleral explant culture system to facilitate the intravitreous delivery of vectors. Posterior segment explants from adult mice of 2 different age groups (4 wk and 15 wk) were cultured in serum-free medium for variable time periods. Tissue viability was assessed by gross morphology, cell survival quantification, activated caspase-3 expression, and immunohistochemistry. To model ocular gene therapy, explants were exposed to varying transducing units of a lentiviral vector expressing the gene for green fluorescent protein for 48 h. Explant retinal cells remained viable for approximately 1 wk, although the ganglion cell layer developed apoptosis between 4 and 7 d. Following vector infusion into the posterior segment cups, viral transduction was noted in multiple retinal layers in both age groups. An age of donor mouse influence was noted and older mice did not transduce as well as younger mice. This explant offers an easily managed posterior segment ocular culture with minimum disturbance of the tissue, and may be useful for investigating methods of enhancing retinal gene therapy under controlled conditions.

OBJECTIVE To evaluate the use of rebound and applanation tonometry for the measurement of intraocular pressure (IOP) and to assess diurnal variations in and the effect of topical anesthesia on the IOP of healthy inland bearded dragons (Pogona vitticeps). ANIMALS 56 bearded dragons from 4 months to 11 years old. PROCEDURES For each animal following an initial ophthalmic examination, 3 IOP measurements were obtained on each eye between 9 AM and 10 AM, 1 PM and 2 PM, and 5 PM and 7 PM by use of rebound and applanation tonometry. An additional measurement was obtained by rebound tonometry for each eye in the evening following the application of a topical anesthetic to evaluate changes in the tolerance of the animals to the tonometer. Descriptive data were generated, and the effects of sex, time of day, and topical anesthesia on IOP were evaluated. RESULTS Bearded dragons did not tolerate applanation tonometry even following topical anesthesia. Median daily IOP as determined by rebound tonometry was 6.16 mm Hg (95% confidence interval, 5.61 to 6.44 mm Hg). The IOP did not differ significantly between the right and left eyes. The IOP was highest in the morning, which indicated that the IOP in this species undergoes diurnal variations. Topical anesthesia did not significantly affect IOP, but it did improve the compliance for all subjects. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that rebound tonometry, but not applanation tonometry, was appropriate for measurement of IOP in bearded dragons. These findings provided preliminary guidelines for IOP measurement and ophthalmic evaluation in bearded dragons.

OBJECTIVE To determine the cardiopulmonary effects of progressively increasing infusion rates of dopamine hydrochloride and phenylephrine hydrochloride in healthy adult New Zealand White rabbits anesthetized with isoflurane. ANIMALS 6 New Zealand White rabbits. (Oryctolagus cuniculus). PROCEDURES Each rabbit was anesthetized on 2 occasions (≥ 2 weeks apart) with isoflurane in oxygen at 1.5 times the published isoflurane minimum alveolar concentration of 2.07%. Carotid artery and pulmonary artery catheters were placed. During each anesthetic episode, each rabbit received 5 progressively increasing doses of either dopamine (5, 10, 15, 20, or 30 μg/kg/min) or phenylephrine (0.125, 0.25, 0.5, 1.0, and 2.0 μg/kg/min). Blood gas and cardiopulmonary measurements were obtained after a 20-minute equilibration period prior to administration of the first drug dose (baseline) and after each subsequent dose administration. RESULTS Dopamine increased stroke index at the highest infusion rate of 30 μg/kg/min; however, cardiac output and mean arterial blood pressure remained unchanged from baseline values. Administration of phenylephrine at a rate of 2 μg/kg/min increased mean arterial blood pressure to 62 mm Hg from the baseline value of 45 mm Hg. This was a result of an increase in systemic vascular resistance with a concomitant decrease in heart rate and no change in cardiac output. Blood lactate concentration increased with time when rabbits received either treatment. CONCLUSIONS AND CLINICAL RELEVANCE Within the dose range of 5 to 30 μg/kg/min, dopamine was not an effective treatment for isoflurane-induced hypotension in rabbits and phenylephrine was only minimally effective at a dose of 2 μg/kg/min.

OBJECTIVE To evaluate whether a low-dosage regimen of prednisolone induces bone loss and whether administration of alendronate sodium prevents glucocorticoid-induced osteopenia in dogs by measuring trabecular bone mineral density (BMD) with quantitative CT. ANIMALS 8 healthy Beagles. PROCEDURES In 4 dogs, prednisolone was administered PO at a dosage of 2 mg/kg once daily for 2 weeks, 1 mg/kg once daily for 4 weeks, and 0.5 mg/kg once daily for 3 weeks. In the other 4 dogs, alendronate sodium (2 mg/kg, PO, q 24 h) was whether administered for 9 weeks in addition to the same dosage of prednisolone used in the prednisolone-treated dogs. Before (day 0 [baseline]) and 21, 42, 63, and 150 days after the start of treatment, BMD of the lumbar vertebrae was measured by quantitative CT. RESULTS BMD in the prednisolone treatment group decreased to 84.7% of the baseline value on day 42, increased to 87.9% on day 63, and recovered to 91.6% on day 150. In the prednisolone-alendronate treatment group, BMD decreased to 91% of the baseline value on day 21, increased to 93.8% on day 63, and then recovered to 96.7% on day 150. Bone mineral density in the prednisolone treatment group was generally lower, albeit not significantly, than that of the prednisolone-alendronate treatment group on each examination day. CONCLUSIONS AND CLINICAL RELEVANCE BMD temporarily decreased after low-dosage prednisolone administration; however, it gradually improved during tapering of the prednisolone dosage. These results have suggested that a low dosage of prednisolone can be used with little concern for development of osteopenia in dogs.