OBJECTIVE To compare the efficacy of application of an alcohol-based antiseptic (80% ethyl alcohol) hand rub (ABAHR) with that of a 2% chlorhexidine gluconate scrub (CGS2) for immediate reduction of the bacterial population on the skin of dogs. ANIMALS 50 client-owned dogs with no evidence of skin disease. PROCEDURES On each dog, 2 areas of hair on the ventral aspect of the abdomen were clipped with a No. 40 blade and cleared of debris. A direct contact plate holding tryptic soy agar with polysorbate 80 and lecithin was gently pressed (for 2 seconds) on each skin site (preapplication sample). The CGS2 and ABAHR were each aseptically applied to 1 skin site on each dog. A direct contact plate was subsequently applied to each site in a similar manner (postapplication sample). All plates were cultured, and bacterial isolates were identified and quantified by the number of CFUs per plate. RESULTS Application of the CGS2 and ABAHR significantly decreased skin bacterial colony counts, compared with findings for preapplication samples. The number of CFUs per plate or postapplication percentage reduction in CFUs per plate did not differ between treatments. There were no adverse skin reactions associated with either application. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that applications of ABAHR and CGS2 were equally effective at immediately reducing the bacterial population on the skin of dogs, and there was no significant difference in percentage reduction in colony counts between the 2 applications.

OBJECTIVE To evaluate the microbial integrity of preservative-free cyclodextrin-based alfaxalone in a multiple-use system. SAMPLE 22 vials of preservative-free alfaxalone. PROCEDURES 2 storage conditions (room temperature, 22°C; refrigerated temperature, 4°C) and 3 handling techniques (closed system transfer device, nonclosed dispensing pin, and manufacturer-supplied vial stopper) comprised 6 treatment groups (3 replicates/group). An aliquot (0.5 mL) was withdrawn from each vial daily for 14 days. Samples were immediately inoculated into tryptic soy broth and incubated at 36°C for 24 hours; samples were subcultured onto 5% Columbia sheep blood agar and incubated for 48 hours. Isolated colonies were evaluated for identification. RESULTS There was no evidence of microbial contamination of vials stored for 7 days in refrigeration and handled with a protected port (closed system transfer device or nonclosed dispensing pin). CONCLUSIONS AND CLINICAL RELEVANCE The US FDA prohibits the use of alfaxalone beyond 6 hours after the vial stopper is broached (punctured), as mandated for a preservative-free injectable medication. Findings for the study reported here supported the use of alfaxalone for 7 days when refrigerated and handled with a single puncture of the stopper by use of a protected port (closed system transfer device or nonclosed dispensing pin). This would appear to be a practical alternative for an injectable anesthetic. It would minimize drug waste and the subsequent environmental impact for disposal of unused drug and allow standardization of storage and handling protocols for alfaxalone use in veterinary practices across the United States.

OBJECTIVE To evaluate gene transfer of recombinant adeno-associated viral (rAAV) vectors with AAV2 or AAV5 capsid and encoding hyaluronic acid (HA) synthase-2 (HAS2) into joints of healthy dogs. ANIMALS 22 purpose-bred Beagles. PROCEDURES Plasmid expression cassettes encoding canine HAS2 (cHAS2) were assessed in vitro for concentration and molecular size of secreted HA. Thereafter, rAAV2-cHAS2 vectors at 3 concentrations and rAAV5-cHAS2 vectors at 1 concentration were each administered intra-articularly into the left stifle joint of 5 dogs; 2 dogs received PBS solution instead. Synovial fluid HA concentration and serum and synovial fluid titers of neutralizing antibodies against AAV capsids were measured at various points. Dogs were euthanized 28 days after treatment, and cartilage and synovium samples were collected for vector DNA and mRNA quantification and histologic examination. RESULTS Cell transfection with plasmids encoding cHAS2 resulted in an increase in production and secretion of HA in vitro. In vivo, the rAAV5-cHAS2 vector yielded uniform genome transfer and cHAS2 expression in collected synovium and cartilage samples. In contrast, rAAV2-cHAS2 vectors were detected inconsistently in synovium and cartilage samples and failed to produce clear dose-related responses. Histologic examination revealed minimal synovial inflammation in joints injected with rAAV vectors. Neutralizing antibodies against AAV capsids were detected in serum and synovial fluid samples from all vector-treated dogs. CONCLUSIONS AND CLINICAL RELEVANCE rAAV5-mediated transfer of the gene for cHAS2 into healthy joints of dogs by intra-articular injection appeared safe and resulted in vector-derived cHAS2 production by synoviocytes and chondrocytes. Whether this treatment may increase HA production by synoviocytes and chondrocytes in osteoarthritic joints remains to be determined.

OBJECTIVE To evaluate the diagnostic yield of dental radiography (Rad method) and 3 cone-beam CT (CBCT) methods for the identification of predefined anatomic landmarks in brachycephalic dogs. ANIMALS 19 client-owned brachycephalic dogs admitted for evaluation and treatment of dental disease. PROCEDURES 26 predefined anatomic landmarks were evaluated separately by use of the RAD method and 3 CBCT software modules (serial CBCT slices and custom cross sections, tridimensional rendering, and reconstructed panoramic views). A semiquantitative scoring system was used, and mean scores were calculated for each anatomic landmark and imaging method. The Friedman test was used to evaluate values for significant differences in diagnostic yield. For values that were significant, the Wilcoxon signed rank test was used with the Bonferroni-Holm multiple comparison adjustment to determine significant differences among each of the 6 possible pairs of diagnostic methods. RESULTS Differences of diagnostic yield among the Rad and 3 CBCT methods were significant for 19 of 26 anatomic landmarks. For these landmarks, Rad scores were significantly higher than scores for reconstructed panoramic views for 4 of 19 anatomic landmarks, but Rad scores were significantly lower than scores for reconstructed panoramic views for 8 anatomic landmarks, tridimensional rendering for 18 anatomic landmarks, and serial CBCT slices and custom cross sections for all 19 anatomic landmarks. CONCLUSIONS AND CLINICAL RELEVANCE CBCT methods were better suited than dental radiography for the identification of anatomic landmarks in brachycephalic dogs. Results of this study can serve as a basis for CBCT evaluation of dental disorders in brachycephalic dogs.

OBJECTIVE To assess and compare 2 injection techniques for conducting ocular anterior segment indocyanine green angiography (ASICGA) and sodium fluorescein (SF) angiography in horses. ANIMALS 3 healthy adult female horses (age range, 19 to 25 years). PROCEDURES Horses were sedated, jugular catheters were placed, and manual restraint was used to ensure proper positioning for the angiography procedure. Two injection techniques (IV and intra-arterial) were performed for each horse 1 week apart. Intravenous injections of 0.25% indocyanine green (ICG; 50 mg) and 10% SF (10 mg/kg) were administered via the jugular catheter. Intra-arterial injections of ICG (1 mg) and SF (1 mg/kg) were administered into the common carotid artery with ultrasound guidance. Angiography was performed by use of an adaptor system comprised of a modified digital single-lens reflex camera, camera adaptor, and lens. Imaging was performed at a rate of 3 images/s for 60 seconds immediately following ICG injection, then at 2, 3, 4, and 5 minutes after injection. The SF was injected 5 minutes thereafter. RESULTS ASICGA allowed visual identification of the arterial, capillary, and venous phases of angiography. Intra-arterial administration provided superior dye fluorescence, sharper contrast, and faster dye passage than IV administration. Visibility of the iris vasculature was limited with SF, and extravasation of SF was noted. No clinically important adverse events were detected. CONCLUSIONS AND CLINICAL RELEVANCE ASICGA images were obtainable with both injection techniques; however, visibility of the iris vasculature was better with intra-arterial administration of ICG. The ASICGA technique may serve as a viable ocular imaging modality for horses.

OBJECTIVE To characterize the MRI and histologic features of the supraspinatus tendon in nonlame dogs. ANIMALS 7 cadavers (14 shoulder joints) of nonlame 2-year-old sexually intact male Beagles. PROCEDURES Multiple MRI fluid-sensitive pulse sequences were obtained for both shoulder joints of each cadaver, and the thickness, volume, and signal intensity of each supraspinatus tendon were assessed. After MRI scanning was complete, the shoulder joints were processed for histologic examination. Tissue specimens were stained with various stains to determine tendon morphology and composition. Histologic and MRI findings were correlated and described. RESULTS All supraspinatus tendons had a trilaminar appearance on sagittal and transverse MRI images, which was characterized by a thick, hyperintense center layer (central substance) sandwiched between thin hypointense superficial and deep margins. The mean ± SD central substance-to-superficial margin and central substance-to-deep margin thickness ratios were 8.4 ± 1.2 and 9.0 ± 0.9, respectively; supraspinatus tendon-to-triceps brachii muscle signal intensity ratio was 1.3 ± 0.2; and tendon volume was 445 ± 20 mm3. The superficial and deep margins histologically resembled other tendons with highly ordered collagen fibers. The central substance was comprised of water-rich glycosaminoglycans interspersed among haphazardly arranged collagen bundles. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated histologically normal canine supraspinatus tendons have a trilaminar appearance on MRI images. In dogs, a diagnosis of supraspinatus tendinosis should not be based solely on the tendon having a hyperintense signal on MRI images; other MRI evidence of shoulder joint disease and diagnostic findings are necessary to support such a diagnosis.

Salmonella enterica serovar Typhimurium has a wide host range and is capable of causing infections ranging from severe gastroenteritis to systemic infection in humans. To determine if attenuated S. Typhimurium strains can serve as safe and effective oral vaccines to prevent typhoid fever, the biologic characteristics of crp and sipB deletion mutants were evaluated. Previous studies had found that the crp and sipB genes are related to Salmonella pathogenicity. In this study, cytotoxicity, protective efficacy, and immune responses of the host were analyzed. Our previous data had shown a significance decrease in virulence for the crp and sipB mutants compared with a wild-type strain. The current study confirmed this finding in HeLa cells and showed that the crp mutant was significantly less cytotoxic (P < 0.05) than the sipB mutant. Mice vaccinated with the crp mutant showed significantly better protection after challenge with the wild-type strain (P < 0.05) and significantly greater responses in serum IgG (P < 0.01) and secretory IgA (P < 0.05) compared with the mice vaccinated with the sipB mutant (P < 0.05). Our results indicate that the crp mutant has the potential to be a vaccine candidate and is safe in mice.

To obtain immunogenic conjugate antigens, adipic acid dihydrazide (ADH), as a bridge, and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimidehydrochloride (EDAC), as a coupling agent, were used to conjugate the purified fusion protein, clumping factor A-fibronectin binding protein ClfA A-FnBPA, and type 5 capsular polysaccharide (CP5). The conjugates were mixed with an adjuvant, and mice were immunized 3 times and challenged with Staphylococcus aureus 1 week later. Antibody titers were determined by indirect enzyme-linked immunosorbent assay (ELISA). At 14 days after the first immunization, antibodies against the purified protein and conjugate were detected; after 28 days, antibody levels increased; and a week after the third immunization, antibody levels continued to increase. However, the conjugate antibody titers were higher than those of the purified protein during the study, and no IgG antibodies against purified CP5 were detected during the entire experiment. The protection rate increased to 90% in the conjugate group, indicating that the conjugate imparts a relatively higher protective efficacy than the purified protein and purified CP5.

OBJECTIVE To investigate water intake and urine measures in healthy cats provided free-choice access to a nutrient-enriched water with (NWP) or without (NW) added poultry flavoring offered at 3 different volumes in addition to tap water (TW). ANIMALS 36 domestic shorthair cats. PROCEDURES Control group cats (n = 4) received dry food with TW ad libitum throughout the study. Cats of the NW and NWP groups (n = 16/group) received the same food with TW only (period 1; 7 days) followed by TW and the assigned treatment ad libitum at 1X, 1.5X, and 2X the volume of TW consumed in period 1 during periods 2 (17 days), 3 (10 days), and 4 (10 days), respectively. Liquid consumption, food intake, and total water intake (from all sources) were measured; urine collected over 48 hours in each period was measured, and urine specific gravity (USG) was determined. Data were analyzed with mixed-effects models. RESULTS TW and food calorie intake were similar among groups in period 1; TW consumption by control cats did not differ during the study. Liquid consumed by drinking increased 18%, 57%, and 96% for the NWP group in periods 2, 3, and 4, respectively, with increases of 25% and 44% for the NW group in periods 3 and 4, respectively, compared with period 1 values for the same groups. Increased urine output and decreased USG were significantly associated with period and treatment. CONCLUSIONS AND CLINICAL RELEVANCE Increasing the volumes of NW or NWP offered to healthy cats led to increased free liquid consumption and was associated with greater urine output and dilution as measured by USG. Studies are warranted to determine whether these treatments provide health benefits for cats in need of greater water consumption.

OBJECTIVE To evaluate cell-mediated and humoral immune responses of calves receiving 2 doses of a dual-adjuvanted vaccine containing inactivated bovine herpesvirus type 1 (BHV1) and bovine viral diarrhea virus types 1 (BVDV1) and 2 (BVDV2) before and after exposure to BHV1. ANIMALS 24 Holstein steers negative for anti-BHV1 antibodies and proliferative cell-mediated immune responses against BHV1 and BVDV. PROCEDURES Calves were randomly assigned to 3 groups. The vaccinated group (n = 10) received 2 doses of vaccine on days 0 and 21. Control (n = 10) and seeder (4) groups remained unvaccinated. Calves were commingled during the study except for the 3-day period (days 53 to 55) when seeders were inoculated with BHV1 (1.04 × 107 TCID50, IV) to serve as a source of virus for challenge (days 56 through 84). Rectal temperature and clinical illness scores were monitored, and blood and nasal specimens were obtained for determination of clinicopathologic and immunologic variables. RESULTS After BHV1 challenge, mean rectal temperature and clinical illness scores were lower for vaccinates than controls. In vaccinates, antibody titers against BHV1 and BVDV2, but not BVDV1, increased after challenge as did extracellular and intracellular interferon-γ expression, indicating a T helper 1 memory response. Additional results of cell marker expression were variable, with no significant increase or decrease associated with treatment. CONCLUSIONS AND CLINICAL RELEVANCE Calves administered 2 doses of a killed-virus vaccine developed cell-mediated and humoral immune responses to BHV1 and BVDV, which were protective against disease when those calves were subsequently exposed to BHV1.