Immunohistochemistry has been used extensively to evaluate protein expression in clinical and research settings. However, immunohistochemistry is not always successful in veterinary medicine due to the lack of reliable antibody options, poor tissue preservation, labor-intensive staining, and antigen-retrieval optimization processes. RNAscope in-situ hybridization (ISH) is a powerful technology that uses a specific sequence probe to identify targeted mRNA. In this study, we demonstrate RNAscope ISH in 4 common canine malignancies, which are traditionally diagnosed by histopathology and immunohistochemistry. Probes were designed for commonly targeted mRNA markers of neoplastic tumors; these included c-kit in mast cell tumor, microphthalmia-associated transcription factor in malignant melanoma, ionized calcium-binding adapter molecule(-1) in histiocytic sarcoma, and alkaline phosphatase in osteosarcoma. A strong staining signal was obtained by these 4 targets in each canine malignancy. These results support the use of RNAscope ISH for definitive diagnosis in canine malignancies.

OBJECTIVE To evaluate the effects of 3 electrolyte solutions administered SC to experimentally dehydrated inland bearded dragons (Pogona vitticeps). ANIMALS 9 inland bearded dragons. PROCEDURES In a randomized, complete crossover study, experimental dehydration was induced by means of furosemide (10 mg/kg, SC, q 12 h for 4 doses), and then lactated Ringer solution, Plasma-Lyte A, or reptile Ringer solution (RRS; 1:1 mixture of 5% dextrose solution and isotonic crystalloid solution) was administered SC in a single 50-mL/kg dose in 3 treatments sessions separated by a minimum of 14 days. Food and water were withheld during treatment sessions. Plasma biochemical values, PCV, blood total solids and lactate concentrations, and plasma osmolarity were measured prior to (baseline) and 4 and 24 hours after fluid administration. RESULTS Administration of RRS resulted in severe hyperglycemia (mean ± SD plasma glucose concentration, 420 ± 62 mg/dL), compared with baseline values (190 ± 32 mg/dL), and this hyperglycemia persisted for at least 24 hours. It also resulted in significant reductions in plasma osmolarity and sodium and phosphorus concentrations, which were not observed after administration of the other 2 solutions. Administration of lactated Ringer solution caused no significant increase in blood lactate concentration. CONCLUSIONS AND CLINICAL RELEVANCE The changes in plasma glucose, sodium, and phosphorus concentrations and plasma osmolarity observed after SC administration of a single dose of RRS suggested this type of electrolyte solution should not be used for rehydration of bearded dragons. Rather, lactated Ringer solution or Plasma-Lyte A should be considered instead.

OBJECTIVE: To assess the analgesic efficacy of an IV constant rate infusion (CRI) of a morphine-lidocaine-ketamine (MLK) combination in calves undergoing umbilical herniorrhaphy. ANIMALS: 20 weaned Holstein calves with umbilical hernias. PROCEDURES: Calves were randomly assigned to receive a CRI of an MLK solution (0.11 mL/kg/h; morphine, 4.8 μg/kg/h; lidocaine, 2.1 mg/kg/h; and ketamine, 0.42 mg/kg/h) for 24 hours (MLK group) or 2 doses of flunixin meglumine (1.1 mg/kg, IV, q 24 h) and a CRI of saline (0.9% NaCl) solution (0.11 mL/kg/h) for 24 hours (control group). The assigned CRI was begun after anesthesia induction. A pain-scoring system and incisional algometry were used to assess pain, and blood samples were obtained to measure serum cortisol concentration at predetermined times for 120 hours after CRI initiation. RESULTS: Mean pain scores did not differ significantly between the MLK and control groups at any time. Mean algometry score for the MLK group was significantly greater (calves were less responsive to pressure) than that for the control group at 4 hours after CRI initiation. Mean cortisol concentration decreased over time for both groups and was significantly greater for the MLK group than the control group at 1, 4, and 18 hours after CRI initiation. CONCLUSIONS AND CLINICAL RELEVANCE: A CRI of MLK provided adequate postoperative analgesia to calves that underwent umbilical herniorrhaphy. However, the technical support required for CRI administration limits its use to hospital settings. Kinetic analyses of MLK infusions in cattle are necessary to establish optimal dosing protocols and withdrawal intervals.

The present study describes the serum biochemical alterations and histopathological abnormalities in the liver tissue of dogs with liver disease. A survey of hepatic lesions was conducted using ultrasound-guided percutaneous needle biopsies. The hematologic and biochemical changes in dogs with liver lesions were recorded. Chronic hepatitis was the most common liver histopathologic finding (37.9%). Other common findings included liver fibrosis (19.5%), vacuolar hepatopathy (10.3%), cholangiohepatitis (9.2%), hepatocellular carcinoma (4.6%), cholangitis (3.4%), cholangiocarcinoma (2.3%), and congestive hepatopathy (2.3%). Greater than 2-fold elevation of serum alanine aminotransferase (ALT) was a useful indicator, with a sensitivity of 40% to 65% for diagnosing all liver pathologies. Greater than 2-fold elevation of serum alkaline phosphatase (ALP) without significant elevation of ALT was useful for diagnosing liver diseases affected by inflammatory or regressive changes (sensitivity of 40% to 50%). Elevation of ALT, ALP, or a combination of ALT and ALP had a high sensitivity of up to 90% for identifying dogs with liver pathology. Hepatic injury and cholestasis enzymes should be interpreted together with patient history, clinical signs, and liver ultrasonographic appearance before performing a liver biopsy.

OBJECTIVE To determine whether jejunal serosal patches could securely seal large, open defects in duodenal segments harvested from canine cadavers and to compare intraluminal pressures at which leakage first occurred and maximal intraluminal pressures for repaired duodenal segments between 2 suture patterns. SAMPLE Duodenal and jejunal segments from 9 canine cadavers. PROCEDURES 20 constructs were created through repair of large, open duodenal defects with circumferential suturing of an intact jejunal segment (jejunal serosal patch). Constructs were randomly assigned to have the serosal patch anastomosed to the duodenal segment by a simple continuous or simple interrupted suture pattern. The pressure at which the first leakage was observed and the maximum pressure obtained during testing were recorded and compared between suture patterns. RESULTS Initial leakage pressure was significantly higher with the simple interrupted pattern (mean ± SD, 68.89 ± 5.62 mm Hg), compared with the simple continuous pattern (59.8 ± 20.03 mm Hg). Maximum intraluminal pressures did not significantly differ between the simple interrupted (91 ± 8.27 mm Hg) and simple continuous patterns (90.7 ± 16.91 mm Hg). All constructs, regardless of suture pattern, withstood supraphysiologic pressures. CONCLUSIONS AND CLINICAL RELEVANCE Jejunal serosal patches adequately sealed large, open duodenal defects and prevented leakage in these constructs. Constructs with simple continuous or simple interrupted suture patterns withstood physiologic and supraphysiologic intraluminal pressures, although constructs with a simple interrupted suture pattern initially leaked at higher pressures.

OBJECTIVE To establish a method for evaluation of the efficacy of a classical swine fever virus (CSFV) subunit vaccine in rabbits as determined via humoral immune responses to the virus. ANIMALS 40 specific pathogen–free rabbits. PROCEDURES Rabbits were randomly assigned to 4 groups (10 rabbits/group) for SC injection of 0.05, 0.1, and 0.2 mL of a CSFV subunit E2 vaccine (representing 1.15, 2.3, or 4.6 μg of E2 protein/dose, respectively) or saline (0.9% NaCl) solution. Blood samples were collected 21 days after vaccination for measurement of the antibody response against CSFV via ELISA and virus neutralization methods. On the same day, the CSFV Chinese (C) strain was injected into an ear vein. Vaccine efficacy was determined by monitoring of rabbits for pyrexia for 4 days and measurement of viral copies in spleen lysates at the end of the study. Reproducibility of the antibody response was tested with 2 other batches of the vaccine at the minimum immunization dose identified for the initially tested batch. RESULTS The E2 protein dose of the initially tested vaccine was positively correlated with the antibody response and protection rate in rabbits. The identified minimum immunization dose per rabbit was 0.1 mL, representing an E2 protein content of approximately 2.3 μg, and reproducibility of the antibody response to vaccination with the 2 other batches at this dose was good. CONCLUSIONS AND CLINICAL RELEVANCE A method was established in rabbits for evaluation of the efficacy of a CSFV subunit vaccine that could help in the optimization of later large-scale vaccine production and quality control processes as well as in the clinical application of the vaccine.

OBJECTIVE To determine the pharmacokinetics of danofloxacin following IM administration of a single dose (10 mg/kg) in koi (Cyprinus carpio). ANIMALS 69 healthy adult koi housed in a 980-L flow-through-system tank. PROCEDURES 3 fish were kept as untreated controls, and the remaining 66 fish were assigned to 11 treatment groups with 6 fish/group. Fish in the treatment groups were given a single dose of danofloxacin (10 mg/kg) IM in the left epaxial musculature. Fifteen, 30, and 45 minutes and 1, 4, 12, 24, 72, 96, 120, and 144 hours after administration of danofloxacin, fish in each treatment group were euthanized, and blood samples and samples of liver, spleen, gill, anterior kidney, posterior kidney, skin and muscle, and scales were collected. Plasma and tissue drug concentrations were determined by liquid chromatography–tandem mass spectrometry, and noncompartmental pharmacokinetic analyses were performed. Tissues from the untreated control fish and fish euthanized 144 hours after danofloxacin administration were examined histologically. RESULTS Maximum plasma concentration (mean, 8,315.7 ng/mL) was reached approximately 45 minutes after danofloxacin administration; plasma elimination half-life was 15 hours. Danofloxacin was detected in all examined tissues from all 6 fish euthanized 15 minutes after drug administration and was detected in some tissues from 3 of the 6 fish euthanized 144 hours after drug administration. For all tissues, results of histologic examination were unremarkable. CONCLUSIONS AND CLINICAL RELEVANCE IM administration of a single dose (10 mg/kg) of danofloxacin in koi resulted in rapid absorption, with maximum plasma concentration reached approximately 45 minutes after drug administration; the drug could still be detected in some tissues 144 hours after administration.

OBJECTIVE To determine perioperative analgesia associated with oral administration of a novel methadone-fluconazole-naltrexone formulation in dogs undergoing routine ovariohysterectomy. ANIMALS 43 healthy female dogs. PROCEDURES Dogs were randomly assigned to receive the methadone-fluconazole-naltrexone formulation at 1 of 2 dosages (0.5 mg/kg, 2.5 mg/kg, and 0.125 mg/kg, respectively, or 1.0 mg/kg, 5.0 mg/kg, and 0.25 mg/kg, respectively, PO, q 12 h, starting the evening before surgery; n = 15 each) or methadone alone (0.5 mg/kg, SC, q 4 h starting the morning of surgery; 13). Dogs were sedated with acepromazine, and anesthesia was induced with propofol and maintained with isoflurane. A standard ovariohysterectomy was performed by experienced surgeons. Sedation and pain severity (determined with the Glasgow Composite Pain Scale-short form [GCPS-SF]) were scored for 48 hours after surgery. Rescue analgesia was to be provided if the GCPS-SF score was > 6. Dogs also received carprofen starting the day after surgery. RESULTS None of the dogs required rescue analgesia. The highest recorded GCPS-SF score was 4. A significant difference in GCPS-SF score among groups was identified at 6:30 am the day after surgery, but not at any other time. The most common adverse effect was perioperative vomiting, which occurred in 11 of the 43 dogs. CONCLUSIONS AND CLINICAL RELEVANCE Oral administration of a methadone-fluconazole-naltrexone formulation at either of 2 dosages every 12 hours (3 total doses) was as effective as SC administration of methadone alone every 4 hours (4 total doses) in dogs undergoing routine ovariohysterectomy. Incorporation of naltrexone in the novel formulation may provide a deterrent to human opioid abuse or misuse.

A study was conducted to investigate the seroprevalence and associated risk factors of Rift Valley fever (RVF) infection in cattle and some selected wildlife species at selected interface areas at the periphery of the Great Limpopo Transfrontier Conservation Area in Zimbabwe. Three study sites were selected based on the type of livestock-wildlife interface: porous livestock-wildlife interface (unrestricted); non-porous livestock-wildlife interface (restricted by fencing) and livestock-wildlife non-interface (totally absent contact or control). Sera were collected from cattle aged &#8805; 2 years representing both female and intact male. Sera were also collected from selected wild ungulates from Mabalauta (porous interface) and Chipinda Pools (non-interface) areas of the Gonarezhou National Park. Sera were tested for antibodies to Rift Valley fever virus (RVFV) using a competitive enzyme-linked immunosorbent assay (ELISA) test. AX2 test was used to assess differences between categories, and p < 0.05 was considered as significant. In cattle, the overall seroprevalence was 1.7% (17/1011) (95% confidence interval &#91;CI&#93;: 1.01-2.7). The porous interface recorded a seroprevalence of 2.3% (95% CI: 1.2-4.3), the non-porous interface recorded a prevalence of 1.8% (95% CI: 0.7-4.3) and the non-interface area recorded a seroprevalence of 0.4% (955 CI: 0.02-2.5), but the difference in seroprevalence according to site was not significant (p &gt; 0.05). All impala and kudu samples tested negative. The overall seroprevalence in buffaloes was 11.7% (95% CI: 6.6-19.5), and there was no significant (p = 0.38) difference between the sites (Mabalauta, 4.4% &#91;95% CI: 0.2-24&#93; vs. Chipinda, 13.6% &#91;95% CI: 7.6-23&#93;). The overall seroprevalence in buffaloes (11.7%, 13/111) was significantly (p < 0.0001) higher than in cattle (1.7%, 17/1011). The results established the presence of RVFV in cattle and selected wildlife and that sylvatic infections may be present in buffalo populations. Further studies are required to investigate if the virus is circulating between cattle and wildlife.