OBJECTIVE To measure expression of microRNAs (miRNAs) in plasma and in extracellular vesicles (EVs) derived from plasma for dogs with glioma and dogs with other brain diseases. SAMPLE Plasma samples from 11 dogs with glioma and 19 control dogs with various other brain diseases. PROCEDURES EVs were isolated from plasma samples by means of ultracentrifugation. Expression of 4 candidate reference miRNAs (let-7a, miR-16, miR-26a, and miR-103) and 4 candidate target miRNAs (miR-15b, miR-21, miR-155, and miR-342-3p) was quantified with reverse transcription PCR assays. Three software programs were used to select the most suitable reference miRNAs from among the 4 candidate reference miRNAs. Expression of the 4 target miRNAs was then calculated relative to expression of the reference genes in plasma and EVs, and relative expression was compared between dogs with glioma and control dogs with other brain diseases. RESULTS The most suitable reference miRNAs were miR-16 for plasma and let-7a for EVs. Relative expression of miR-15b in plasma and in EVs was significantly higher in dogs with glioma than in control dogs. Relative expression of miR-342-3p in EVs was significantly higher in dogs with glioma than in control dogs. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that miR-15b and miR-342-3p have potential as noninvasive biomarkers for differentiating glioma from other intracranial diseases in dogs. However, more extensive analysis of expression in specific glioma subtypes and grades, compared with expression in more defined control populations, will be necessary to assess their clinical relevance.

OBJECTIVE To determine whether cell-free DNA (cfDNA) was detectable in CSF samples from dogs, whether CSF sample volume impacted CSF cfDNA concentration measurement, and whether CSF cfDNA concentration was associated with CNS disease category or CSF RBC count (RBCC), nucleated cell count (NCC), or protein concentration, which could aid in the diagnosis of neurologic diseases in dogs. SAMPLE 80 CSF samples collected from dogs with (n = 60) and without (20) clinical neurologic disease between February 2017 and May 2018. PROCEDURES Results for CSF RBCC, NCC, protein concentration, and cfDNA concentration were compared across CSF groups established on the basis of whether they were obtained from dogs with (case groups) or without (control group) clinical signs of neurologic disease In addition, 5 paired CSF samples representing large (3.0-mL) and small (0.5-mL) volumes, were used to evaluate whether sample volume impacted measurement of CSF cfDNA concentration. RESULTS cfDNA was detected in 76 of the 80 (95%) CSF samples used to evaluate parameters across disease categories and in all 5 of the paired samples used to evaluate whether sample volume impacted cfDNA quantification. There were no substantial differences in cfDNA concentrations identified between groups (on the basis of disease category or sample volume), and the CSF cfDNA concentration did not meaningfully correlate with CSF RBCC, NCC, or protein concentration. CONCLUSIONS AND CLINICAL RELEVANCE Although results indicated that the CSF cfDNA concentration could not be used to differentiate between categories of neurologic disease in dogs of the the present study, further investigation is warranted regarding the use of CSF analysis, including sequencing specific cfDNA mutations, for diagnosing and monitoring neurologic disease in dogs.

OBJECTIVE To evaluate effects of topical ophthalmic administration of diclofenac on intraocular pressure (IOP) when applied at 4 frequencies to eyes of Beagles. ANIMALS 8 ophthalmologically normal Beagles. PROCEDURES The study involved four 5-day experimental periods each separated by a 16-day washout period. During each period, 1 drop of 0.1% diclofenac sodium ophthalmic solution was administered to the right eye at 4 treatment frequencies (1, 2, 3, or 4 times/d); 1 drop of eyewash was administered to the left eye as a control treatment. A complete ophthalmic examination was performed on days 0 (day before first treatment) and 5 of each experimental period. Gonioscopy was performed on day 0 of the first period. The IOPs were measured at 7 am and 7 pm on days 1 through 5. RESULTS No abnormalities were detected during neuro-ophthalmic and ophthalmic examinations on day 0 of each experimental period. No adverse reactions to administration of diclofenac or eyewash were observed at any time point. No abnormalities were detected during ophthalmic examinations performed on day 5, and IOPs remained < 25 mm Hg in all 4 periods. No significant differences were identified between the treated and control eyes or among the 4 treatment frequencies. CONCLUSIONS AND CLINICAL RELEVANCE Topical ophthalmic administration of diclofenac up to 4 times/d in dogs with no ophthalmic abnormalities did not significantly increase the IOP. Additional research is needed to evaluate the effect of topical ophthalmic administration of diclofenac on IOP in dogs with anterior uveitis.

OBJECTIVE To determine the feasibility of sidestream dark field (SDF) video microscopy for the evaluation of the jejunal microvasculature of healthy dogs. ANIMALS 30 healthy sexually intact female shelter dogs anesthetized for ovariohysterectomy. PROCEDURES Preoperative physical and clinicopathologic assessments were performed to confirm health status. Then healthy dogs were anesthetized, and the abdomen was incised at the ventral midline for ovariohysterectomy and jejunal microvasculature evaluation. An SDF video microscope imaged the microvasculature of 2 sites of a portion of the jejunum, and recorded videos were analyzed with software capable of quantitating parameters of microvascular health. Macrovascular parameters (heart rate, respiratory rate, and hemoglobin oxygen saturation) were also recorded during anesthesia. RESULTS Quantified jejunal microvascular parameters included valid microvascular density (mean ± SD, 251.72 ± 97.10 μm/mm), RBC-filling percentage (66.96 ± 8.00%), RBC column width (7.11 ± 0.72 μm), and perfused boundary region (2.17 ± 0.42 μm). The perfused boundary region and RBC-filling percentage had a significant negative correlation. Strong to weak positive correlations were noted among the perfused boundary regions of small-, medium-, and large-sized microvessels. No significant correlations were identified between microvascular parameters and age, body weight, preoperative clinicopathologic results, or macrovascular parameters. CONCLUSIONS AND CLINICAL RELEVANCE Interrogation of the jejunal microvasculature of healthy dogs with SDF video microscopy was feasible. Results of this study indicated that SDF video microscopy is worth additional investigation, including interrogation of diseased small intestine in dogs.

OBJECTIVE To investigate the activities of gelatinases (matrix metalloproteinase [MMP]-2 and MMP-9) and serine proteases in the colorectal mucosa of Miniature Dachshunds (MDs) with inflammatory colorectal polyps (ICRPs). ANIMALS 15 MDs with ICRPs and 5 dogs with non–ICRP-related large bowel diarrhea (controls). PROCEDURES Zymographic methods were used to evaluate the activities of MMP-2, MMP-9, latent forms of MMP-2 and MMP-9 (pro–MMP-2 and pro–MMP-9), and serine proteases in inflamed and noninflamed tissue samples from MDs with ICRPs and in noninflamed tissue samples from control dogs. The associations of serine protease activities with MMP-2 or MMP-9 activity were also analyzed. RESULTS Activities of pro–MMP-2 and pro–MMP-9 were detected in most tissue samples, regardless of the tissue type, whereas activities of MMP-2 and MMP-9 were not detected in control tissue samples. In the inflamed tissue samples from MDs with ICRPs, the activities of MMP-2, pro–MMP-9, and MMP-9 were significantly higher than those in the noninflamed tissue samples from those dogs. Serine protease activities were significantly higher in the inflamed and noninflamed tissue samples from MDs with ICRP, compared with findings for control tissue samples. A weak correlation was detected between serine protease activities and MMP-9 activity. CONCLUSIONS AND CLINICAL RELEVANCE Study results suggested that gelatinase and serine protease activities are upregulated in the colorectal mucosa of MDs with ICRPs, possibly contributing to the pathogenesis of this disease through the functions of these enzymes in degradation of extracellular matrix and promotion of inflammatory cell migration and inflammatory responses.

OBJECTIVE To assess clotting times, coagulation factor activities, sterility, and thromboelastographic parameters of liquid plasma (LP), thawed fresh frozen plasma (FFP-T), and 2 novel formulations of freeze-dried plasma (FDP) stored refrigerated over 35 days. SAMPLE 6 units of canine LP and FFP-T from a commercial animal blood bank and 5 units each of 2 formulations of canine FDP. PROCEDURES Prothrombin time; activated partial thromboplastin time; activities of coagulation factors II, V, VII, VIII, IX, X, XI, and XII; and thromboelastographic parameters were determined for each product on days 0 (baseline), 3, 7, 14, 21, 28, and 35. For each day, a sample of each product was also submitted for aerobic bacterial culture. RESULTS Small changes in coagulation factor activities and mild increased time to initial clot formation in LP and FFP-T were noted over the 35-day storage period. Activities of factor VIII in FDP1 and factor XII in FDP2 were < 50% at baseline but varied throughout. Compared with FFP-T, time to initial clot formation was increased and clot strength was preserved or increased for the FDPs throughout the study. One FDP had decreased pH, compared with other products. No plasma product yielded bacterial growth. CONCLUSIONS AND CLINICAL RELEVANCE Liquid plasma and FFP-T would be reasonable to use when stored refrigerated for up to 35 days. Both FDP products showed variability in coagulation factor activities. Studies investigating the usefulness of these plasma products (FDPs) in dogs and the variable days of refrigerated storage (all products) are warranted.

OBJECTIVE To determine the effects of coadministration of naltrexone, a human opioid abuse deterrent, on the pharmacokinetics and pharmacodynamics of a methadone-fluconazole combination administered orally to dogs. Animals: 12 healthy Beagles. PROCEDURES Dogs (body weight, 10.7 to 13.9 kg) were randomly allocated to 2 groups in a parallel design study. All dogs received fluconazole (100 mg [7.19 to 9.35 mg/kg], PO). Twelve hours later (time 0), dogs were administered methadone (10 mg [0.72 to 0.93 mg/kg]) plus fluconazole (50 mg [3.62 to 4.22 mg/kg]; methadone-fluconazole) or methadone (10 mg [0.72 to 0.93 mg/kg]) plus fluconazole (50 mg [3.60 to 4.67 mg/kg]) and naltrexone (2.5 mg [0.18 to 0.23 mg/kg]; methadone-fluconazole-naltrexone), PO, in a gelatin capsule. Blood samples were collected for pharmacokinetic analysis, and rectal temperature and sedation were assessed to evaluate opioid effects at predetermined times up to 24 hours after treatment. RESULTS Most dogs had slight sedation during the 12 hours after drug administration; 1 dog/group had moderate sedation at 1 time point. Mean rectal temperatures decreased significantly from baseline (immediate pretreatment) values from 2 to ≥ 12 hours and 2 to ≥ 8 hours after methadone-fluconazole and methadone-fluconazole-naltrexone treatment, respectively. Geometric mean maximum observed concentration of methadone in plasma was 35.1 and 33.5 ng/mL and geometric mean terminal half-life was 7.92 and 7.09 hours after methadone-fluconazole and methadone-fluconazole-naltrexone treatment, respectively. Naltrexone was sporadically detected in 1 dog. The active naltrexone metabolite, β-naltrexol, was not detected. The inactive metabolite, naltrexone glucuronide, was detected in all dogs administered methadone-fluconazole-naltrexone. CONCLUSIONS AND CLINICAL RELEVANCE Opioid effects were detected after oral administration of methadone-fluconazole or methadone-fluconazole-naltrexone. Further studies assessing additional opioid effects, including antinociception, are needed.

Sarcoptic mange is a pruritic, contagious, ectoparasitic skin disease that affects mammals, including the domestic dog. The objective of this study was to evaluate and compare the efficacy of afoxolaner plus milbemycin oxime (NexGard Spectra) and afoxolaner alone (NexGard) as treatments for sarcoptic mange in naturally infested dogs. A total of 142 dogs naturally infested with Sarcoptes scabiei was evaluated. The dogs were diagnosed by microscopic examinations of skin scrapings. The dogs were divided into 2 groups: 96 dogs were treated with a combined dosage of 2.50 to 5.36 mg/kg body weight (BW) of afoxolaner and 0.50 to 1.07 mg/kg BW of milbemycin oxime and 46 dogs were treated with 2.50 mg/kg BW of afoxolaner alone. The presence or absence of pruritus and lesions were evaluated using an analogous scale on days 7, 14, 21, 28, and 56 after receiving the treatment. Data obtained were analyzed by Student's t-test (P ≤ 0.05). The single oral treatment of afoxolaner plus milbemycin oxime resulted in a significant reduction in pruritus of 87.4% at 28 d after treatment (P ≤ 0.05). Resolution of the lesions after treatment was variable, with a significant decrease (P ≤ 0.05) observed within the first 14 d, although this parameter continued to improve until the end of the study on day 28, when a decrease of 96% was observed. By the end of the study, a single dose of either the afoxolaner alone or the afoxolaner combined with milbemycin oxime was effective in significantly reducing the signs associated with sarcoptic mange during a 56-day evaluation period.

OBJECTIVE To determine the thermal antinociceptive effects of butorphanol tartrate and butorphanol tartrate in a sustained-release 25% poloxamer 407 (P407) gel formulation (But-P407) in parrots. ANIMALS 13 orange-winged Amazon parrots (Amazona amazonica). PROCEDURES First, butorphanol tartrate (5 mg/kg) or saline (0.9% NaCl) solution was administered IM to birds in a randomized complete crossover design. The temperature prompting a foot withdrawal response to a thermal stimulus (ie, the thermal threshold) was determined 30 minutes before (baseline) and at various points after treatment administration. Second, But-P407 (12.5 mg/kg) or P407 was administered SC in a similar crossover design. Thermal threshold was determined before and at various points after treatment administration. Third, But-P407 (12.5 mg/kg) or saline solution was administered SC and evaluated as in the second trial. Sedation was scored immediately before each time point in all 3 trials. RESULTS In the first trial, a significant increase in thermal threshold was noted 30 minutes after butorphanol tartrate (vs saline solution) administration. No sedation was noted. In the second and third trials, no significant difference was identified between results for But-P407 and those for either control treatment (saline solution or P407). Mild sedation was noted in the second trial following But-P407 administration. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested a small but significant thermal antinociceptive effect of butorphanol tartrate lasting between 30 minutes and 1.5 hours in orange-winged Amazon parrots. No antinociceptive effect of butorphanol tartrate was demonstrated when delivered in P407. Further research is needed to evaluate the potential analgesic effects of But-P407.

Recent studies have found that anemia and anisocytosis are precipitating factors for certain heart diseases in dogs. This study evaluated the prevalence and correlation of anemia and red blood cell distribution width (RDW) in dogs with heartworm disease (HWD). The study population consisted of 20 healthy control dogs and 86 dogs with HWD: 28 dogs with no clinical signs or pulmonary hypertension (Group 1), 42 dogs with mild clinical signs but no pulmonary hypertension (Group 2), and 16 dogs with severe clinical signs and pulmonary hypertension (Group 3). Along with echocardiographic interrogation of pulmonary hypertension, red blood cell (RBC) profiles were evaluated, including RDW. The total number of red blood cells (tRBCs), hematocrit (HCT), and hemoglobin (HGB) concentration was significantly lower in Group 3 dogs compared to control dogs (P < 0.05), while the RDW was significantly higher in Group 3 dogs than in control dogs (P < 0.05). The RDW was closely correlated to other RBC profiles and the presence of pulmonary hypertension (P < 0.05). The severity of tricuspid regurgitant gradient (TRG) was closely correlated with Hb and tRBC (P < 0.05), but not with the RDW and reticulocyte count. This study finding indicated that anemia and anisocytosis are common complications in dogs with severe clinical signs and pulmonary hypertension caused by heartworm disease (HWD). It would therefore be beneficial for clinicians to routinely check red blood cell (RBC) profiles, including RDW, in order to monitor the progression of heartworm disease in dogs.