Kinetic analysis of demethylation of 13C-aminopyrine in healthy dogs

OBJECTIVE: To describe the kinetics of demethylation of 13C-aminopyrine in healthy dogs for use in determining the most appropriate time for collection of blood samples for a 13C-aminopyrine demethylation blood test for evaluation of hepatic function. ANIMALS: 9 healthy dogs. PROCEDURES: A 2-mL baseline blood sample was collected into an evacuated heparinized tube, and 13C-aminopyrine was administered to each dog (2 mg/kg, IV). Additional 2-mL blood samples were collected 15, 30, 45, 60, 75, 90, 105, 120, 135, 150, 180, 240, 300, and 360 minutes after 13C-aminopyrine administration. The CO2 was extracted from blood samples by addition of a strong acid, and the percentage dose of 13CO2 (PCD) in the extracted gas was determined by fractional mass spectrometry. RESULTS: No dogs had gross evidence of adverse effects, and all had an increase in PCD after IV administration of 13C-aminopyrine. The PCD had the least variability among 5 variables used to evaluate hepatic demethylating capacity. Peak PCD was detected at 30 minutes in 1 dog, 45 minutes in 5 dogs, 60 minutes in 2 dogs, and 75 minutes in 1 dog. The mean PCD for the 9 dogs peaked at 45 minutes after 13C-aminopyrine administration. CONCLUSIONS AND CLINICAL RELEVANCE: PCD appears to be the preferable variable for evaluation of hepatic demethylating capacity. Intravenous administration of 13C-aminopyrine leads to a consistent increase in PCD. Mean PCD peaked 45 minutes after administration, suggesting that blood sample collection 45 minutes after 13C-aminopyrine administration may be appropriate for use in estimating hepatic demethylating capacity.     

Comparison of various doses of carbon 13-labeled aminopyrine for a carbon 13-labeled aminopyrine demethylation blood test in healthy dogs

OBJECTIVE: To determine an optimal dose of carbon 13 ((13)C)-labeled aminopyrine for use in a (13)C-aminopyrine demethylation blood test in healthy dogs. ANIMALS: 9 adult dogs. PROCEDURES: Food was withheld from each dog for 12 hours. A 2-mL baseline blood sample was obtained from each dog and placed into an evacuated tube containing sodium heparin. Carbon 13-labeled aminopyrine was administered IV at doses of 1, 2, 5, or 10 mg/kg. Additional blood samples (2 mL) were obtained and placed into evacuated tubes containing sodium heparin 30, 45, 60, and 75 minutes after (13)C-aminopyrine administration. Hydrochloric acid was used to extract CO(2) from blood samples. The extracted gas was analyzed by fractional mass spectrometry to determine the percentage dose of (13)C administered as (13)C-aminopyrine and recovered in extracted gas (PCD). RESULTS: Gross evidence of clinical adverse effects was not detected in any dog after administration of (13)C-aminopyrine. The mean coefficient of variation (CV) for PCD was significantly lower than the mean CV for the summation of PCD values up to a given sampling time (CUMPCD). Mean PCD values among the 4 doses for each sample time were not significantly different. Administration of (13)C-aminopyrine at a dose of 2 mg/kg resulted in the lowest interindividual variability. CONCLUSIONS AND CLINICAL RELEVANCE: The PCD is superior to CUMPCD for the quantification of aminopyrine demethylation. Administration of (13)C-(13)C-aminopyrine at a dose of 2 mg/kg is appropriate for use in the (13)C-aminopyrine demethylation blood test in healthy dogs.     

Use of a 13C-aminopyrine blood test: first clinical impressions

The purpose of this study was to collect initial data to determine the potential clinical usefulness of a ¹³C-aminopyrine demethylation blood test, and whether additional clinical investigation is warranted. Six dogs, initially suspected of having hepatic disease based on their history, physical examination, imaging studies, general laboratory parameters, or any combination of the above, were enrolled in the study. A baseline blood sample was collected, 2 mg/kg ¹³C-aminopyrine was administered intravenously, and another blood sample was collected 45 min afterwards. Carbon dioxide was extracted from the blood samples and analyzed using fractional mass spectrometry. Results from the ¹³C-aminopyrine demethylation blood test were compared to clinical data and histologic findings. Intravenous administration of ¹³C-aminopyrine leads to a decrease in the percent dose of ¹³C recovered from dogs with histologically confirmed liver disease. Based on our results, a full-scale investigation of the potential clinical usefulness of a ¹³C-aminopyrine demethylation blood test for assessment of hepatic function in dogs is warranted.     

Development of a 13C-glycocholic acid blood test to assess bacterial metabolic activity of the small intestine in canines

The objectives of this study were to establish optimal doses of 13C-glycocolic acid (GCA) for use in a GCA blood test as a marker for canine small intestinal bacterial metabolic activity. Four doses of GCA were administered orally to 8 healthy dogs. Blood samples were collected at various time points up to 480 min. The percent dose/min of 13C administered as GCA (PCD) and cumulative PCD (CUMPCD) were determined by fractional mass spectrometry. No dog showed any clinically obvious side effects. Doses of 1 and 2 mg/kg of bodyweight (BW) led to a significant increase in PCD and CUMPCD (P < 0.001). The mean CUMPCD was significantly higher for the 1 mg/kg BW dose compared with the 2 and 4 mg/kg BW doses (P < 0.05). Administration of 1 mg/kg BW of 13C-glycocholic acid led to an increase in CUMPCD over baseline in gas extracted from blood samples and appears to be the best parameter to evaluate for future clinical studies.     

Kinetic analysis of 5 sugar probes in dog serum after orogastric administration

The objective of this study was to describe the kinetics of orally administered sugar probes in serum for the assessment of gastrointestinal permeability and intestinal absorptive capacity in dogs. Eight healthy dogs received lactulose (L), rhamnose (R), methylglucose (M), xylose (X), and sucrose (S) by orogastric intubation. Baseline blood samples and subsequently timed blood samples were taken during 24 hours. Sugars were analyzed by gas chromatography-mass spectrometry (GC-MS). Statistical analysis was performed using a Friedman test with Dunn’s multiple comparison post test and a Kruskal-Wallis test. Statistical significance was set at a P-value < 0.05. Sugars in serum were detected after orogastric administration. Concentrations of L and R were significantly different from the baseline from 90 to 240 and 60 to 300 min, respectively, and those of X, M, and S were different from 30 to 240 min post-dosing (P < 0.05 for all 5 probes). Maximum concentrations of L and R were obtained at 180 min, while X, M, and S reached their maximum concentrations at 90 min post-dosing. For all sugars, no statistically significant differences were found between concentrations at 90, 120, and 180 min or between the coefficients of variation (CV%) of those mean concentrations for these 3 time points. Based on these data, the collection of 2 blood samples, one taken at baseline and the other obtained between 90 and 180 minutes after dosing, might be sufficient for the determination of gastrointestinal permeability and mucosal absorptive capacity using these 5 sugar probes in canine serum.     

Effects of preemptive atropine administration on incidence of medetomidine-induced bradycardia in dogs

OBJECTIVE: To determine the cardiorespiratory effects of preemptive atropine administration in dogs sedated with medetomidine. DESIGN: Randomized crossover trial. ANIMALS: 12 healthy adult dogs. PROCEDURES: Dogs underwent 6 treatments. Each treatment consisted of administration of atropine (0.04 mg/kg [0.018 mg/lb] of body weight, IM) or saline solution (0.9% NaCl, 1 ml, IM) and administration of medetomidine (10, 20, or 40 microg/kg [4.5, 9.1, or 18.2 microg/lb], IM) 10 minutes later. Treatments were administered in random order, with a minimum of 1 week between treatments. Cardiorespiratory effects before and after atropine and medetomidine administration were assessed. Duration of lateral recumbency and quality of sedation and recovery were assessed. RESULTS: Bradycardia (heart rate < 60 beats/min) was seen in all dogs when saline solution was administered followed by medetomidine, and the dose of medetomidine was not associated with severity or frequency of bradycardia or second-degree heart block. However, a medetomidine dose-dependent increase in mean and diastolic blood pressures was observed, regardless of whether dogs received saline solution or atropine. Preemptive atropine administration effectively prevented bradycardia and second-degree heart block but induced pulsus alternans and hypertension. The protective effects of atropine against bradycardia lasted 50 minutes. Blood gas values were within reference limits during all treatments and were not significantly different from baseline values. Higher doses of medetomidine resulted in a longer duration of lateral recumbency. CONCLUSIONS AND CLINICAL RELEVANCE: Preemptive administration of atropine in dogs sedated with medetomidine effectively prevents bradycardia for 50 minutes but induces hypertension and pulsus alternans.     

Formation of monoethylglycinexylidide (MEGX) in clinically healthy dogs

Liver function tests help to investigate actual liver function. In dogs, only a few tests are available. We evaluated the formation of monoethylglycinexylidide (MEGX) in clinically healthy dogs to assess the usefulness of this liver function test in dogs. Twenty-five healthy dogs were used in this study. The MEGX test was done according to human protocols. The results of our study showed that dogs synthesize MEGX after the administration of lidocaine. There was no age dependence of this test in dogs and no significant difference between measurements obtained at 15 and 30 min after administration of lidocaine. Female dogs had significantly (P < 0.05) higher concentrations of MEGX 15 min after administration. The reference interval for dogs after 15 min is 34 to 79 μg/L and after 30 min 39 to 89 μg/L. In conclusion, the MEGX test may be an additional liver function test in dogs.     

Acid–Base Changes in Canine Neonates Following Normal Birth or Dystocia

There are limited data concerning blood gas parameters in neonatal dogs. Knowledge of the normal physiology may facilitate effective therapeutic intervention and potentially reduce neonatal mortality. This study examined acid–base parameters in pups born at normal parturition (n = 27) compared with those born after obstetrical assistance or caesarean operation (n = 13) and those born following oxytocin (OXY) administration for treatment of uterine inertia (n = 11). Pups were subjected to an objective scoring method of neonatal health adapted from use in humans (the Apgar score) at birth and again at 5 and 60 min after birth. Venous blood samples were collected at 5 and 60 min after birth for evaluation of blood gas parameters. At birth, all pups had low Apgar scores and a mixed acidosis. The base excess was lowest for pups delivered after OXY administration. The Apgar score improved for all pups after 5 min of birth and there was an improvement in carbon dioxide tension, base excess and venous blood pH at 1 h, although in all pups a metabolic acidosis persisted. These data provide an important insight into neonatal physiology and the variability of blood gas parameters in pups born at normal and abnormal parturition and provide the basis for clinical decision making following dystocia.     

Evaluation of splanchnic oxygen extraction ratio as an index of portal vein pressure in dogs

OBJECTIVE: To determine changes in splanchnic oxygen extraction ratio during experimentally induced portal hypertension in dogs. ANIMALS: 6 clinically normal dogs. PROCEDURE: Standard midline laparotomy and median sternotomy were performed in anesthetized dogs. Baseline measurements of arterial blood pressure, aortic blood flow, portal vein blood flow, and portal vein pressure were acquired, and arterial, venous, and portal vein blood samples were obtained to determine systemic and splanchnic oxygen extraction ratios. The portal vein was gradually occluded until a pressure of 18 cm of H2O was reached; this pressure was maintained for 30 minutes, and measurements and collection of blood samples were repeated. RESULTS: Portal vein blood flow decreased significantly from 457 +/- 136 ml/min before to 266 +/- 83 ml/min after induction of portal hypertension. Oxygen content in the portal vein significantly decreased from 12.3 +/- 1.85 to 8.2 +/- 2.31%, and splanchnic oxygen extraction ratio significantly increased from 15.8 +/- 6.2 to 37.4 +/- 10.9% during portal hypertension. There was a significant inverse correlation between portal vein blood flow and splanchnic oxygen extraction ratio at baseline and during portal hypertension. CONCLUSIONS AND CLINICAL RELEVANCE: An increase in splanchnic oxygen extraction ratio is evident with partial attenuation of the portal vein and the concurrent decrease in portal vein blood flow. Correlation of oxygen extraction ratio with portal vein blood flow may be a more important indicator for determination of an endpoint to prevent congestion and ischemia of the gastrointestinal tract and pancreas during ligation of portosystemic shunts.     

Effects of a peripheral alpha2 adrenergic-receptor antagonist on the hemodynamic changes induced by medetomidine administration in conscious dogs

OBJECTIVE: To evaluate the effects of administration of a peripheral alpha(2)-adrenergic receptor antagonist (L-659,066), with and without concurrent administration of glycopyrrolate, on cardiopulmonary effects of medetomidine administration in dogs. ANIMALS: 6 healthy adult dogs. PROCEDURES: Dogs received saline (0.9% NaCl) solution (saline group), L-659,066 (group L), or L-659,066 with glycopyrrolate (group LG). These pretreatments were followed 10 minutes later by administration of medetomidine in a randomized crossover study. Hemodynamic measurements and arterial and mixed-venous blood samples for blood gas analysis were obtained prior to pretreatment, 5 minutes after pretreatment, and after medetomidine administration at intervals up to 60 minutes. RESULTS: After pretreatment in the L and LG groups, heart rate, cardiac index, and partial pressure of oxygen in mixed-venous blood (PvO2) values were higher than those in the saline group. After medetomidine administration, heart rate, cardiac index, and PvO2 were higher and systemic vascular resistance, mean arterial blood pressure, and central venous pressure were lower in the L and LG groups than in the saline group. When the L and LG groups were compared, heart rate was greater at 5 minutes after medetomidine administration, mean arterial blood pressure was greater at 5 and 15 minutes after medetomidine administration, and central venous pressure was lower during the 60-minute period after medetomidine administration in the LG group. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of L-659,066 prior to administration of medetomidine reduced medetomidine-induced cardiovascular changes in healthy dogs. No advantage was detected with concurrent administration of L-659,066 and glycopyrrolate.     

Effects of oral administration of a commercial activated charcoal suspension on serum osmolality and lactate concentration in the dog

The purpose of this investigation was to determine the effects of an activated charcoal (AC) suspension containing propylene glycol and glycerol on serum osmolality, osmolal gap, and lactate concentration in dogs. Six healthy adult dogs were administered 4 g/kg AC in a commercially available suspension that contained propylene glycol and glycerol as vehicles. Blood samples were taken before and 1, 4, 6, 8, 12, and 24 hours after the administration of the test suspension. Samples were analyzed for osmolality, blood gases, and concentrations of lactate, sodium, potassium, serum urea nitrogen, and glucose. Osmolal gaps were calculated for each time point. Mean serum osmolality, osmolal gap, and lactate concentration were significantly increased after suspension administration compared to baseline. Serum osmolality increased from 311 mOsm/kg at baseline to 353 mOsm/kg, osmolal gap increased from 5 to 52 mOsm/kg, and lactate concentration increased from 1.9 to 4.5 mmol/L after suspension administration (all P < .01). Three of the 6 dogs vomited between 1 and 3 hours after the administration of the test suspension, and 4 of 6 dogs were lethargic. All dogs drank frequently after AC administration. Commercial AC suspension administered at a clinically relevant dose increases serum osmolality, osmolal gap, and lactate concentration in dogs. These laboratory measures and the clinical signs of vomiting, lethargy, and increased frequency of drinking might complicate the diagnosis or monitoring of some intoxications (such as ethylene glycol) in dogs that have previously received AC suspension containing propylene glycol, glycerol, or both as vehicles.     

Acute response of urine pH following ammonium chloride administration to dogs

To test the acidifying ability of the distal portion of the nephrons in healthy dogs, 0.2 g of NH4Cl/kg of body weight was given PO. Samples for venous blood gas analysis and urine pH were taken hourly for 6 hours. Systemic acidemia developed, as evidenced by a statistically significant (P less than 0.05) decrease in blood pH 1 hour after NH4Cl administration. Four hours after administration, mean urine pH decreased to a low of 5.16 +/- 0.1 and was less than 5.5 3 hours after administration. Changes in urine pH 2 hours after administration were statistically significant (P less than 0.05). In human beings, NH4Cl loading is used to detect patients with distal renal tubular acidosis (defective hydrogen ion secretion by the distal nephrons) and normal acid/base values. Distal renal tubular acidosis is diagnosed if urine pH fails to decrease to less than 5.5 after NH4Cl administration. On the basis of the findings of this study, a similar value would be valid for dogs.     

Evaluation of a hand-held lactate analyzer in dogs

A hand-held lactate test device and a blood gas auto analyzer were compared. The objective of the study was to evaluate the performance of the hand-held device in dogs in a clinical setting. Blood lactate levels were evaluated on 30 samples from healthy client-owned dogs and 48 samples from client-owned dogs with various diseases. A blood sample was collected from each healthy dog by either jugular or cephalic venipuncture and from each sick dog from the jugular, cephalic, or saphenous vein, or from an arterial catheter if applicable. One and a half milliliters of the blood sample was immediately transferred to a heparinized vacutainer tube. Enough blood was then drawn from the heparinized tube to allow split sample simultaneous analysis with both machines. Samples from the sick dogs represented a wide range of clinically relevant lactate values. Good agreement between lactate values from both devices was obtained in both sick and healthy dogs. Lactate values in the healthy group (< 2.9 mmol/L with the hand-held device, < 2.6 mmol/L with the blood gas analyzer) were similar to those previously reported (< 2.5 mmol/L). The results of this study support the use of the hand-held device in dogs in a clinical setting.     

Erythrocyte and plasma fatty acid patterns in dogs with atopic dermatitis and healthy dogs in the same household

Recent studies have indicated that dogs with canine atopic dermatitis (CAD) may have a disorder of fatty acid metabolism: possibly low or absent activity of delta6-desaturase or delta5-desaturase, or both. To clarify this possibility, we examined the erythrocyte and plasma fatty acid patterns of 8 dogs with CAD and their 8 healthy housemates. Atopic dermatitis was diagnosed according to the criteria proposed by Willemse; other causes of dermatitis were excluded clinically and by appropriate tests. Erythrocyte ghosts were prepared from blood samples. Membrane lipids were extracted and separated by thin-layer chromatography. From plasma and lipid fractions, fatty acid content was determined by gas chromatography. In erythrocytes, but not in plasma, we observed significant differences in the fatty acid pattern that suggested a reduction in the n6 fatty acid products of the delta6- and delta5-desaturases in dogs with atopic dermatitis when compared with healthy housemates.     

Comparison of plasma histamine levels after intravenous administration of hydromorphone and morphine in dogs

This study compared plasma histamine concentrations, behavioral and cardiovascular parameters following intravenous administration of hydromorphone and morphine in conscious dogs. Five adult female dogs received a 15-sec bolus injection of saline, hydromorphone (0.1 and 0.2 mg/kg) or morphine (0.5 and 1.0 mg/kg) randomly at weekly intervals. Blood samples were collected from the jugular vein before and at 1, 2, 5, 15, 30, 60 and 120 min after drug administration. Plasma histamine concentration, noninvasive oscillometric blood pressure, heart rate and rhythm were evaluated. Data were analyzed with repeated measures anova and Tukey-Kramer post hoc test with a 5% significance level. Median plasma histamine increased significantly only after the higher dose of morphine. Maximum plasma histamine measured was 0.8 ng/mL after saline and, after the lower and higher doses, respectively, 10.2 and 9.7 ng/mL for hydromorphone, and 440 and 589 ng/mL for morphine. One dog became hypotensive immediately after receiving the highest dose of morphine. Occasional ventricular premature contractions occurred in one dog after both opioids and dosages. No dogs vomited or defecated, but all salivated profusely with both opioids. Neuroexcitation occurred in four dogs following each opioid. In conclusion, intravenous hydromorphone induced minimal histamine release and was well tolerated by these conscious healthy dogs.     

Sonographic features of drug-induced splenic congestion

Drug-induced splenic congestion has been reported in dogs secondary to barbiturate administration. This research attempted to verify and quantify size changes associated with drug-induced splenic congestion in dogs. Transverse plane ultrasound images of the spleen in normal dogs were collected to determine the maximum diameter in the minimum dimension prior to, and 15 min after, administration of acepromazine, thiopental, or propofol. Significant splenic enlargement was seen after administration of acepromazine (P<0.01) and thiopental (P=0.02), but no enlargement was seen after administration of propofol. Significantly increased attenuation (P<0.01) and a trend of increased backscatter (P=0.09) were measured after administration of acepromazine. These results indicate that measurable splenomegaly occurs after acepromazine and thiopental administration. This represents the first report of a condition causing measurable diffuse increased attenuation in the spleen. Propofol does not cause measurable splenic enlargement under the conditions of this research.     

Experimental acute toxicity of xylitol in dogs

The Cases of xylitol poisoning in dogs are increasing as a result of ingestion of xylitol-containing products. Eighteen adult, clinically normal Pekingese dogs were orally dosed with 1 or 4 g/kg xylitol in aqueous solution. Blood samples were collected before and after dosing. Plasma insulin concentrations of both treated groups rose sharply from 20 min after xylitol dosing, peaking at 40 min. Hypoglycemia followed the increase in insulin concentration, with blood glucose values started to decrease 30 min after dosing. Other plasma biochemistry changes associated with xylitol administration were increased alanine aminotransferase and aspartate aminotransferase activities, hypophosphatemia, hypokalemia, and hypercalcemia. Plasma sodium and chloride concentrations remained normal. This study established a biochemical basis for diagnosis and treatment of xylitol poisoning in dogs.     

Development of hypertension and effects of benazepril hydrochloride in a canine remnant kidney model of chronic renal failure

In order to determine whether hypertension would develop in dogs with chronic renal failure, we performed 7/8 renal ablation in 6 healthy dogs and compared pre- and post-ablation blood pressures determined by telemetry. One month after the renal ablation, blood urea nitrogen and creatinine were significantly increased (p<0.05), creatinine clearance was decreased (p<0.05), and blood pressure was increased significantly (p<0.05). Simultaneously, plasma renin activity, angiotensin I and II, and aldosterone were elevated significantly (p<0.05) compared with the values obtained from 11 healthy dogs with intact renal function. The dogs with induced renal failure and hypertension were administered an angiotensin-converting enzyme inhibitor, benazepril hydrochloride, once daily for 2 weeks at 2 mg/kg body weight, and changes in blood pressure and the renin-angiotensin-aldosterone (RAA) system were determined. During the administration of benazepril hydrochloride, blood pressure, angiotensin II and aldosterone decreased significantly (p<0.05) and, upon discontinuation of administration, increased to the pre-administration levels (p<0.05). Plasma renin activity and angiotensin I showed no significant changes throughout the administration study. These results provide experimental evidence that hypertension develops in dogs with chronic renal failure through mechanisms involving the RAA system and demonstrate that benazepril hydrochloride improves renal hypertension in dogs.     

Cardiopulmonary effects of epidurally administered xylazine in the horse

This study was designed to determine whether the epidural administration of an alpha2 agonist, xylazine, would produce measurable changes in arterial blood pressure, electrocardiographic (ECG) activity and arterial blood gas values in horses. Six horses were given each of four treatments: epidural xylazine, intravenous xylazine, epidural lidocaine and epidural saline. A carotid artery catheter was used to measure arterial blood pressure and to collect samples for blood gas analysis before treatment and at intervals post treatment. Heart rate, arterial pressures, ECG activity and respiratory rate were recorded at the same intervals. No significant changes were recorded between time intervals or between individual treatments. It was concluded that this method of xylazine administration to horses produced potent caudal analgesia without measurable cardiopulmonary effects.     

Effect of cimetidine on pharmacokinetics of orally administered cyclosporine in healthy dogs.

OBJECTIVE: To describe the pharmacokinetics of cyclosporine (CyA) in healthy dogs after oral administration alone or in combination with orally administered cimetidine. ANIMALS: 10 healthy adult Beagles. PROCEDURE: Dogs were randomly assigned to receive CyA alone or CyA in combination with cimetidine. After a washout period of 2 weeks, dogs then received the alternate treatment. The CyA plus cimetidine treatment required administration of cimetidine (15 mg/kg of body weight, PO, q 8 h) for 8 days and administration of CyA (5 mg/kg, PO, q 24 h) on days 6 through 8. The CyA treatment alone required administration of CyA (5 mg/kg, PO, q 24 h) for 3 days. On the third day of CyA administration during each treatment, blood samples were collected immediately before (time 0) and 0.5, 1, 1.5, 2, 2.5, 3, 5, 7, 9, 11, 13, 15, 21, and 24 hours after initiating CyA administration. RESULTS: Time until maximum CyA concentration was significantly longer for CyA in combination with cimetidine. Assessment of estimated pharmacokinetic variables revealed a significantly faster rate of change in the distribution phase for CyA in combination with cimetidine. Maximum CyA concentration differed significantly among dogs but did not differ significantly between treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of our data suggests that cimetidine may affect absorption of orally administered CyA, but overall, it does not affect the pharmacokinetics of CyA. There is considerable variability in the maximum concentration of CyA among dogs, and monitoring of blood concentrations of CyA during treatment is advised.