Comparison of urine fibrinogen and interleukin-6 concentrations between healthy dogs and dogs with risk factors for enterococcal bacteriuria.

OBJECTIVE: To compare urine concentrations of fibrinogen (uFIB) and interleukin-6 (uIL-6) between dogs with risk factors for enterococcal bacteriuria and healthy dogs. SAMPLE: Banked urine samples with negative aerobic culture results from 8 dogs with urolithiasis, 9 dogs with anatomic abnormalities of the lower portion of the urinary tract (LUT), 10 dogs with LUT neoplasia, and 21 healthy control dogs. PROCEDURES: Urine creatinine concentration (uCrea) was determined by an automated biochemical analyzer, and uFIB and uIL-6 were determined by dog-specific ELISAs. The uFIB:uCrea and uIL-6:uCrea ratios were calculated for each sample to normalize intersample differences in urine concentration and were compared among the 4 experimental groups. RESULTS: Median uFIB:uCrea ratios for dogs with urolithiasis (0.72; interquartile [25th to 75 percentile] range [IQR], 0.46 to 3.48) and LUT neoplasia (6.16; IQR, 3.89 to 12.75), but not for dogs with LUT anatomic abnormalities (0.48; IQR, 0.27 to 0.69), were significantly greater than that for control dogs (0.17; IQR, 0.07 to 0.39). Median uIL-6: uCrea ratios for dogs with urolithiasis (0.48; IQR, 0.18 to 1.61), LUT anatomic abnormalities (0.25; IQR, 0.17 to 0.33), and LUT neoplasia (0.25; IQR, 0.12 to 1.01) were significantly greater than that for control dogs (0.08; IQR, 0.06 to 0.11). CONCLUSIONS AND CLINICAL RELEVANCE: The uFIB and uIL-6 in dogs with risk factors for enterococcal bacteriuria were generally greater than corresponding values in control dogs. Further investigation is necessary to determine the role of fibrinogen in enterococcal colonization of the urinary tract of dogs.

Comparison of commercial manual extraction kits for RNA isolation from canine whole blood.

High quantities of quality RNA are necessary for many veterinary laboratory tests. Several commercial kits are available for RNA isolation from human whole blood; their resultant RNA yield and purity have not been reported for canine whole blood, to our knowledge. We assessed the performance of 4 RNA extraction kits (RiboPure, TRIzol, RNeasy Protect animal blood, and QIAamp RNA blood mini). Whole blood from a healthy dog was stored in the manufacturer-recommended RNA stabilizing buffer as directed. RNA isolation, including DNase treatment, was performed using each kit's manufacturer's protocol. Resultant RNA yield and purity were evaluated using spectrophotometric absorbance, capillary electrophoresis and electropherogram analysis, and a reverse-transcription real-time PCR (RT-rtPCR) assay. The RNeasy Protect animal blood kit extracted the highest, and RiboPure the lowest, concentration of nucleic acid. RNA integrity numbers classified extracted RNA as good quality or better for all kits except RNeasy Protect. All kits had evidence of genomic DNA contamination as assessed by RT-rtPCR. Overall, QIAamp RNA blood mini kit and TRIzol optimized both RNA yield and purity from canine whole blood. These kits extracted high quantities of good quality RNA as evidenced by high RNA integrity numbers and minimal contamination with proteins and solvents.