Topically administered tissue plasminogen activator (tPA) was evaluated for its penetration into aqueous humor of clinically normal dogs. Two concentrations of tPA (5 mg/ml and 10 mg/ml) were evaluated in a single-dose study, and a concentration of 5 mg of tPA/ml was used for a multiple-dose study. The contralateral eye served as a nontreated control. Enzyme substrate analysis of aqueous humor was used to determine tPA activity. The activity of tPA in aqueous humor was significantly (P < 0.05) greater in treated eyes of all dogs, compared with that in control eyes. Significant differences in activity of tPA were not detected at different doses in treated eyes.

Seven mature dairy cows from 6 herds were obtained with history, clinical signs of disease, and laboratory findings suggestive of advanced paratuberculosis. A surgically implanted collection chamber was used to obtain peripheral tissue fluid. Blood, mammary gland flush fluid, and collection chamber flush fluid (CCFF) samples were obtained 6 times over a 2-week period from each cow. Mononuclear cell-rich portions of these fluids obtained by gradient centrifugation were submitted for bacteriologic culture of Mycobacterium paratuberculosis and for total and differential cell counts. Bacteriologic culture of feces for M paratuberculosis and complete necropsy performed on each cow at the conclusion of the study confirmed the diagnosis of paratuberculosis. Numbers of tissue macrophages obtained from CCFF samples were lower than expected. Mean (+/- SD) differential count of tissue macrophages collected from CCFF was 65.57 (+/- 23.39). Mean calculated tissue macrophages (total cell count X differential count) collected from CCFF samples was 623.1 (+/- 784.55) cells/microliter. Mycobacterium paratuberculosis was isolated from 1 of 42 (2.4%) collections of mononuclear cell-rich portions of plasma and from 2 of 42 (4.8%) CCFF samples. Mycobacterium paratuberculosis was not isolated from any collections of mammary gland flush fluid. The collection and processing techniques used in this study did not enhance detection of M paratuberculosis infection in cows with advanced paratuberculosis, beyond that of ileocecal lymph node biopsy or fecal culture.

In 6 anesthetized ponies, 3 segments of jejunum and 3 segments of small colon were isolated from the peritoneal cavity in plastic bags filled with Hanks' balanced salt solution. One jejunal and 1 small colon segment were subjected to venous strangulation obstruction for 3 hours (VSO-3), venous strangulation obstruction for 6 hours (VSO-6), or a 6-hour sham procedure to control for changes induced by isolation in a plastic bag. Additional segments of jejunum and colon that were not placed in bags served as controls for histologic examination and collagenase measurements. Samples of fluid surrounding the intestine were obtained for chemical analyses, nucleated cell count, aerobic and anaerobic bacteriologic culture, and measurement of collagenase activity. Full-thickness tissue samples were obtained for histologic examination and measurement of collagenase content. Bacteria did not cross the intestinal wall after 3 and 6 hours of VSO, despite severe mucosal lesions in these segments. At 6 hours, P(O2) was significantly less and P(CO2) was significantly (P < 0.05) greater in the fluid surrounding the VSO-6 jejunal segments, compared with the sham jejunal segments. The pH was significantly (P < 0.05) less in fluid surrounding VSO-6 small colon segments, compared with the sham colon segments at 6 hours. For jejunum and small colon, phosphate and lactate concentrations were significantly (P < 0.05) greater in VSO-6 fluid than in the corresponding sham fluids at 6 hours. Fibrin formed around all VSO segments, although fibrinogen was not detected in the surrounding fluid, indicating possible rapid conversion of fibrinogen to fibrin. Fluid collagenase activity increased significantly (P < 0.05) in all segments over 6 hours. The preparation used in this study was successful in measuring local changes on the serosal surface of intestine subjected to VSO and in isolating segments under study in a sterile environment.

Clinically normal dogs were evaluated in states of dehydration, euhydration, and after fluid administration to determine effects of hydration state on renal clearance values. Endogenous creatinine, exogenous creatinine, and [(14)C]inulin clearances, were determined to measure glomerular filtration rate (GFR); in some experiments p-aminohippurate clearance was determined to measure renal plasma flow. Dehydration caused significant (P < 0.05) decrease in clearance values, compared with euhydration, and clearance values during euhydration were significantly (P < 0.05) less than values obtained after a single gavage with water (30 ml/kg of body weight). Sustained administration of 3 fluid regimens was evaluated for effects on clearance values (treatment A = 30 ml of lactated Ringer's solution/kg/h; treatment B = 30 ml of water/kg by gavage hourly; treatment C = 10 ml of glucose:lactated Ringer's solution/ kg/h). All regimens of fluid therapy caused significant P < 0.05), progressive increases in GFR, but treatment C resulted in the most stable GFR values. Increases in clearance values were associated with positive fluid balance; the rate of fluid administration was greater than the rate of urine formation. Data from 285 GFR determinations on 85 dogs were evaluated retrospectively. For each determination, three 20-minute urine collections were made beginning 40 minutes after 30 mi of water/kg was given by gavage. Values between collections were significantly (P < 0.05) different, but varied by < 3%. Comparison of methods for measurement of GFR indicated that endogenous creatinine clearance and [14)C]inulin clearance were highly correlated (R(2) = 0.82), but mean clearance values were markedly different (mean +/- SEM, 28.70 +/- 0.01 and 37.07 +/- 1.29 ml/min, respectively). Exogenous creatinine clearance and [(14)C]inulin clearance were highly correlated (R(2) = 0.95), and mean values were 40.54 +/- 0.70 and 41.02 +/- 0.70 ml/min respectively. We conclude that: state of hydration has a marked effect on GFR; rate of fluid administration that exceeds rate of urine production results in progressive increases in GFR; a single water gavage of 30 ml/kg gives stable GFR values for three 20-minute collection periods, may avoid subclinical states of dehydration, and facilitates accurate urine collections; and endogenous creatinine clearance, as conducted in this study, does not accurately measure GFR.