Estimation of quantitative proteinuria in the dog, using the urine protein-to-creatinine ratio from a random, voided sample

The correlation between 24-hour urine protein excretion and the protein-to-creatinine ratio (U-P/C) from random, voided urine specimens was assessed in 16 healthy Beagles (9 to 11 months old) and in 14 dogs with suspected renal proteinuria. Initially, a voided urine specimen was obtained from each dog, and the U-P/C was determined. An attempt was not made to standardize the time of collection of the voided urine. Subsequently, each dog was placed in a metabolism cage, and 24-hour urine specimens were collected for quantitative protein analysis. The Coomassie blue technique was used to measure urine protein. The correlation between the U-P/C and the 24-hour urine protein excretion (mg/kg/24 hr), evaluated by linear-regression analysis, was found to be significant (r = 0.975, P less than 0.01). These results substantiate previous findings and indicate that random, voided urine specimens may be used to compute the ratio and to accurately reflect 24-hour urinary protein loss in the dog.     

Creatinine in the dog: a review

Creatinine is the analyte most frequently measured in human and veterinary clinical chemistry laboratories as an indirect measure of glomerular filtration rate (GFR). Although creatinine metabolism and the difficulties of creatinine measurement have been reviewed in human medicine, similar reviews are lacking in veterinary medicine. The aim of this review is to summarize information and data about creatinine metabolism, measurement, and diagnostic significance in the dog. Plasma creatinine originates from the degradation of creatine and creatine phosphate, which are present mainly in muscle and in food. Creatinine is cleared by glomerular filtration with negligible renal secretion and extrarenal metabolism, and its clearance is a good estimate of GFR. Plasma and urine creatinine measurements are based on the nonspecific Jaffé reaction or specific enzymatic reactions; lack of assay accuracy precludes proper interlaboratory comparison of results. Preanalytical factors such as age and breed can have an impact on plasma creatinine (P-creatinine) concentration, while many intraindividual factors of variation have little effect. Dehydration and drugs mainly affect P-creatinine concentration in dogs by decreasing GFR. P-creatinine is increased in renal failure, whatever its cause, and correlates with a decrease in GFR according to a curvilinear relationship, such that P-creatinine is insensitive for detecting moderate decreases of GFR or for monitoring progression of GFR in dogs with severely reduced kidney function. Low sensitivity can be obviated by determining endogenous or exogenous clearance rates of creatinine. A technique for determining plasma clearance following IV bolus injection of exogenous creatinine and subsequent serial measurement of P-creatinine does not require urine collection and with additional studies may become an established technique for creatinine clearance in dogs.     

Effects of collection time and food consumption on the urine protein/creatinine ratio in the dog

Effects of collection time and food consumption on the variability of the urine protein/creatinine ratio were determined in 10 healthy dogs. In trial 1, dogs were fasted for 12 hours, and urine specimens were obtained by bladder catheterization every 2 hours over an 8-hour collection period during the day. After a 1-week rest, the dogs were entered into trial 2. Dogs were fed at least 60 kcal of a high protein meal/kg of body weight, and urine specimens were obtained every 2 hours over an 8-hour period during the day. Urine total protein and urine creatinine concentrations and the urine protein/creatinine ratio were determined for each urine specimen obtained. Friedman's 2-way analysis by ranks was used to determine the constancy of this ratio over the 4 periods in the 2 trials (fasted and fed). There was no significant variability (P greater than 0.05) in ratios over the 8-hour collection periods in the fasted or fed trial. Feeding did not significantly alter this ratio, because there was no significant difference (P greater than 0.05) in the urine protein/creatinine ratios of the dogs when they were fasted, compared with those of the dogs when they were fed. Seemingly, urine specimens obtained anytime during the day from dogs in both trials (fasted and fed) reflected the urine protein/creatinine ratio.     

Measurement of aldosterone in feline, canine and human urine

BACKGROUND: Systemic hypertension is an important problem in older cats associated with kidney disease and hypokalaemia, suggesting that excessive activity of the renin-angiotensin-aldosterone system might contribute to the hypertensive state. Fluctuations in plasma renin activity and plasma aldosterone concentrations complicate the interpretation of these assays. OBJECTIVES: The aim of this study was to determine whether measurement of urinary aldosterone excretion in cats aided the investigation of hypertension. METHODS: Urine concentrations of free (ethyl acetate extract) and 18-glucuronidated aldosterone (acid hydrolysis before extraction) were measured by radioimmunoassay in normal, normotensive and hypertensive azotaemic cats (n=11 per group). Urine samples from 11 healthy human volunteers and eight normal dogs were also analysed for comparison. Urinary aldosterone concentration was corrected for the urinary creatinine concentration. RESULTS: Cats excreted 7.3 times less free aldosterone than human beings, and no free aldosterone was detected in dog urine. Acid hydrolysis led to large increases in aldosterone recovery from both human beings and dog but not feline urine. No significant effect of hypertension or azotaemia on feline urinary aldosterone concentration was found. CLINICAL SIGNIFICANCE: Measurement of aldosterone in feline urine using the available methodology has limited or no utility in investigating feline hypertension.     

Effect of storage time on the urine protein: creatinine ratio in alkaline ovine urine

The urine protein:creatinine (UPC) ratio is considered the reference method to assess proteinuria. Its diagnostic value in ovine medicine needs further elucidation. In population monitoring and/or for research purposes, it is convenient to collect many samples simultaneously and store them for later analysis. However, analyte stability data are required to ensure reliable results. We used 15 of 90 urine samples collected from sheep to assess the effect of storage time on the UPC ratio. After centrifugation, the supernatant of each sample was divided into 6 aliquots. Urine protein and creatinine concentrations were determined immediately in one aliquot using the pyrogallol red and a modified Jaffè method, respectively. The other aliquots were stored at −18°C. Based on the absence of active sediment, alkaline urine pH, and UPC ratio ≥0.2, we included 15 samples in our study. The UPC ratio was determined in the stored aliquots 2, 7, 14, 21, and 60 d after collection. The data were analyzed with univariate ANOVA. No significant difference was observed in the urinary concentrations of protein, creatinine, and the UPC ratio (0.8 ± 0.84 in conventional units and 0.09 ± 0.095 in SI units) among different times (p > 0.05). The UPC ratio remained stable for 2 mo in ovine urine samples stored at −18°C.     

Effect of collection time and exercise restriction on the prediction of urine protein excretion, using urine protein/creatinine ratio in dogs

Nonproteinuric and proteinuric dogs were studied to determine whether the urine protein/creatinine ratio from a 24-hour urine sample could be used to predict urine protein excretion. Urine protein/creatinine ratios estimated from urine produced during daylight hours and from that produced during nighttime hours were compared to determine whether time of sample collection influenced the prediction of the urine protein excretion value. Urine protein/creatinine ratios in urine from male dogs were compared with those from female dogs to determine whether sex had an influence on the value. Hospitalized and nonhospitalized dogs were used to determine the effect of exercise restriction. The urine protein/creatinine ratio varied significantly between healthy and proteinuric dogs (P = 0.0001). It was not influenced by collection period or sex. Animals not confined to hospital cages had a significantly lower urine protein/creatinine ratio than did hospitalized animals confined to a cage (P = 0.003).     

Estimation of urine flow rate in mares

To determine the rate of urine flow and thus urinary excretion in the horse from untimed urine samples alone, the flow rate, creatinine concentration, osmolarity, and refractive index of 228 quantitatively collected urine samples were determined in 53 experiments on 12 healthy Thoroughbred mares. Forty samples were collected after water-induced diuresis; 11 samples were collected after furosemide-induced diuresis. Flow rates, which ranged from 1.2 to 84.5 ml/min, could be predicted from the urinary creatinine concentration. Correlation of urinary flow with urinary creatinine concentration accounted for 94% of the variability in the urinary flow rates. Phenylbutazone was administered before collection of 168 urine samples. Urine flow rates that were predicted from urinary creatinine concentration were used to estimate phenylbutazone excretion. Urine flow could be estimated without quantitative urine collection.     

Creatinine concentration in plasma from dog, rat, and mouse: a comparison of 3 different methods

BACKGROUND: There are numerous methods for analyzing creatinine concentration in plasma, including the Jaffé alkaline picrate method in various modifications, enzymatic tests, and chromatographic methods. OBJECTIVE: The purpose of this study was to evaluate whether an enzymatic method could replace a Jaffé method for routine creatinine measurements in plasma from dogs, rats, and mice. The enzymatic method and a compensated Jaffé method were tested against a high-pressure liquid chromatography (HPLC) method, regarded as the gold standard for creatinine measurement. METHODS: Heparinized plasma samples were obtained from 20 beagle dogs, 20 Wistar rats, and 20 CD1-strain mice. The 2 test kits (Roche Diagnostics), Creatinine Jaffé Compensated and the enzymatic Creatinine Plus Version 2 reagent, were used on a Cobas Integra 400. The Jaffé compensated method used a calibration adjustment of 18 micromol/L to correct for the protein matrix in serum and plasma. The HPLC method was an isocratic method using a weak cation-exchange column following protein precipitation. RESULTS: Creatinine concentrations obtained using the enzymatic and the Jaffé methods differed significantly from the results obtained by the HPLC method. For dog plasma, mean values of 61.2, 61.8, and 67.8 micromol/L were obtained by the compensated Jaffé, enzymatic and HPLC methods, respectively. In the rat, respective mean values were 26.7, 21.9, and 23.0 micromol/L, and in the mouse, respective mean values were 14.2, 5.4, and 9.2 micromol/L. CONCLUSION: The enzymatic method can replace the Jaffé method for plasma creatinine determination in dogs, rats, and mice because results from the enzymatic method were closer to HPLC values than were those of the Jaffé method.     

Estimation of roaming dog populations in Timor Leste

The continued spread of rabies through the eastern islands of Indonesia poses a risk of rabies introduction to Timor Leste. To prepare for such an incursion and to undertake surveillance activities, the size and distribution of the roaming dog population needs to be estimated. We present the results of the first such surveys ever undertaken in Timor Leste.     

Inappropriate secretion of antidiuretic hormone in a dog.

Hyponatremia with simultaneous renal sodium loss was associated with the inappropriate secretion of antidiuretic hormone in a dog with heartworm disease. Antidiuresis caused expansion of extracellular fluid volume, which induced renal salt wasting and a negative sodium balance. The combination of water retention, salt wasting, and inactivation of intracellular solute contributes to the decrease in serum sodium concentration. Water intoxication due to hypotonicity of body gluids induced anorexia, depression, weakness, and incoordination.     

Green discolouration of urine following propofol infusion in a dog

A three‐year‐old, female neutered Weimaraner was presented with a history of neck pain and tetraparesis. MRI revealed an extradural mass at the level of C3 vertebra, which was thought to be a spinal abscess, and the dog was scheduled for surgical exploration the following morning. Overnight the dog developed an exaggerated ventilatory pattern, with paradoxical inward movement of the thorax on inspiration. Arterial blood gas analysis revealed respiratory acidosis and ventilator support was initiated to prevent excessive respiratory fatigue. During mechanical ventilation, anaesthesia was maintained using a propofol target‐controlled infusion system and, subsequently, the dog produced bright green urine in the urine collection system. Although previously documented in humans, this appears to be the first report of green urine in a dog following propofol use.     

Serum and urine inorganic fluoride concentrations and urine oxalate concentrations following methoxyflurane anesthesia in the dog.

Plasma fluoride, urine fluoride and urine oxalate concentrations were measured before administering an anesthetic to 8 dogs, and at 0, 3, 9, 24, 48, and 72 hours following 1.5 hours of anesthesia with 1% methoxyflurane. Plasma and urine osmolalities were measured and compared with fluoride and oxalate values. Fluoride concentration increased in both plasma and urine following anesthesia when compared with the preanesthetic concentrations. Maximum mean plasma inorganic fluoride was 106.71 mumoles per liter (+/- 25.44 SE) at 9 hours after exposure to methoxyflurane was completed. By 72 hours after exposure to methoxyflurane the plasma fluoride concentration was 23.47 microM/L (+/- 5.74 SE). Mean urine inorganic fluoride concentration was highest at 9 hours after exposure to methoxyflurane and reached 6047.03 microM/L (+/- 1378.46 SE) as compared to the mean preanesthetic base-line concentration of 542.68 microM/L (+/- 132.93 SE), and the 72 hour mean urine fluoride concentration which was 1593.78 microM/L (+/- 579.46 SE). Urine oxalate concentrations, when compared with urine osmolality (mg/mOsm), increased throughout the study. The 72-hour concentration after exposure to methoxyflurane was 2.5 times the preanesthetic (mg/mOsm) oxalate concentration. Plasma osmolality did not change markedly during the study. Urine osmolalities varied between animals and collection times, but a consistent pattern did not occur. Clinical and laboratory signs of renal dysfunction were not observed in any animal during the study.     

Comparison of the techniques of evaluation of urine dilution/concentration in the dog

The objective of this study was to evaluate the quality of the measurement of dog urine dilution/concentration by comparing osmolality with three methods of specific gravity (USG) measurement, i.e. weighing, refractometry and test strips. In unselected urine samples from 182 dogs there was a better agreement between osmolality and USG determination by refractometry (r = 0.92) than by weighing (r = 0.82) or by test strips (r = 0.27). There was an almost linear relationship between osmolality and USG: osmolality (mOsm/kg) = 36646(34318/38974) x (USGref - 1) + 25(-39/88); calculated osmolality differed from measured osmolality by more than 500 mOsm/kg in only 8 of 181 samples. There was a good agreement between USG determination by weighing and refractometry: USGref = 1.000(0.905/1.095) x USGweighing - 0.0004(-0.0019/0.0027), with a moderate bias. Only 12% of the differences between the two methods exceeded 0.010. Test strip assessment of USG was unreliable because of systematic underestimation and should not be used for dog urine. Refractometry is the best technique for routine evaluation of urine concentration/dilution when osmometry is not available.