A longitudinal study of the blood and urine metabolome of Vipera berus envenomated dogs.

Envenomation of dogs by the common European adder (Vipera berus) is associated with high morbidity. The cytotoxic venom of Vipera berus contains enzymes with the potential to cause acute kidney injury, among other insults, however robust biomarkers for such effects are lacking. A prospective observational follow-up study of naturally envenomated dogs and controls was conducted to fill knowledge gaps regarding canine Vipera berus envenomation, attempt to identify novel biomarkers of envenomation and related kidney injury, and elucidate potential long-term effects. Blood and urine samples were analyzed with a global metabolomics approach using liquid chromatography-mass spectrometry, uncovering numerous features significantly different between cases and controls. After data processing and feature annotation, eight features in blood and 24 features in urine were investigated in order to elucidate their biological relevance. Several of these are associated with AKI, while some may also originate from disturbed fatty acid ß-oxidation and soft tissue damage. A metabolite found in both blood and a venom reference sample may represent identification of a venom component in case dogs. Our findings suggest that envenomated dogs treated according to current best practice are unlikely to suffer permanent injury.

Comparative genetic and pathogenic approaches of Escherichia coli isolated simultaneously from pyometra and urine of bitches.

Escherichia coli (E. coli) are widely related to pyometra and cystitis in dogs, and these infections can occur simultaneously. The goal of this study was to determine genetic and pathogenic insights of 14 E. coli isolated simultaneously from pyometra content and bladder urine of seven bitches. To achieve this, in silico and in vitro comparative analyses were conducted. Whole-genome comparisons demonstrated that E. coli isolated from pyometra and urine of the same animal were predominantly genetic extraintestinal E. coli clones belonging to the same Sequence Type and phylogroup. The E. coli clones identified in this study included ST372, ST457, ST12, ST127, ST646, and ST961. Five isolates (35.7%) belonged to the ST12 complex. Except for two E. coli, all other isolates belonged to the B2 Clermont phylogroup. Interestingly, some genomes of E. coli from urine carried more virulence genes than those E. coli from pyometra. Both pyometra and urine E. coli isolates demonstrated a strong affinity for adhering to HeLa and T24 cells, with a low affinity for invading them. However, certain isolates from urine exhibited a greater tendency to adhere to T24 cells in qualitative and quantitative assays compared to isolates from pyometra. In conclusion, this study revealed the high genomic similarity between pyometra and urine E. coli isolates, as well as the virulent capacity of both to colonize endometrial and urothelial cells. The findings of this study underscore the importance of concurrently managing both infections clinically and could potentially contribute to future resources for the prevention of cystitis and pyometra.

Urinary and household chemical exposures in pet dogs with urothelial cell carcinoma.

Urothelial cell carcinoma (UCC) has been linked to environmental chemical exposures in people, but these risk factors are not well understood in dogs with UCC. We hypothesised that household chemical exposures contribute to the risk of UCC in pet dogs. This prospective cross-sectional case-control study included 37 dogs with UCC and 37 unaffected breed-, sex-, and age-matched controls. Dog owners completed an environmental questionnaire and household samples were collected and analysed for arsenic (in tap water and room dust) and acrolein (in room air). Urine samples from UCC dogs, control dogs, and consenting owners were analysed for inorganic arsenic species, the acrolein metabolite 3-HPMA, and the phenoxy herbicide 2,4-D. Public data on chlorination byproducts (total trihalomethanes) in municipal drinking water were also compared between case and control households. Dogs with UCC were more likely to swim in a pool (15.2%) compared with control dogs (0%) (OR 1.69, 95% CI = 1.69-∞; p = .02). Dogs with UCC also had more than 4-fold higher reported municipal water concentrations of chlorination byproducts (median 28.0 ppb) compared with controls (median 6.9 ppb; p < .0001). Dust arsenic concentrations were unexpectedly lower in case households (median 0.277 ng/cm2) compared with control households (median 0.401 ng/cm2; p = .0002). Other outcomes were not significantly different between groups. These data suggest that dog owners, especially those of breeds known to be at higher risk for UCC, consider limiting access to swimming pools and installing water filtration units that remove total trihalomethanes.

Florfenicol urinary excretion and its potential for treating canine urinary tract infections.

The canine urinary excretion of florfenicol was evaluated to explore its potential for treating urinary tract infections. Nine healthy male intact purpose-bred Beagles and four healthy client-owned dogs each received a single oral dose of florfenicol 20 mg/kg (300 mg/mL parenteral solution) with food. All voluntary urinations were collected for 12 h. Although florfenicol is reportedly bitter tasting, 7/9 Beagles and 4/4 client-owned dogs completely ingested the florfenicol and were enrolled; salivation (n = 1) and headshaking (n = 3) were observed. The last measured urine florfenicol concentrations were variable: Beagles (0.23-3.19 mcg/mL), Pug (3.01 mcg/mL) English Setter (21.29 mcg/mL), Greyhound (32.68 mcg/mL), and Standard Poodle (13.00 mcg/mL). Urine half-life was similar for the Beagles and the Pug, 0.75-1.39 h, whereas the half-life was 1.70-1.82 h for the English Setter, Greyhound, and Standard Poodle. Larger breed dogs exceeded 8 mcg/mL florfenicol (wild-type cutoff) in their urine at 12 h, whereas the Beagles and Pug had <8 mcg/mL; it is unclear if this is an individual, breed, or size difference. These data suggest oral florfenicol may need to be administered q6-12h for canine urinary tract infections, but further data are needed (more enrolled dogs, multiple-dose regimens) before considering clinical trials or breed-specific differences.

Evaluation of a urine dipstick protein to urine specific gravity ratio for the detection of proteinuria in dogs and cats.

BACKGROUND: The utility of urine dipsticks for the quantification of proteinuria is limited because of the influence of urine specific gravity (USG). To circumvent the need for urine protein creatinine ratios (UPCR) some have proposed a calculated dipstick urine protein to USG ratio (DUR) for the detection of proteinuria. However, the performance of DUR has not been evaluated in veterinary patients. OBJECTIVES: Evaluate the correlation between DUR and UPCR, while also assessing the effect of urine characteristics on this relationship and evaluating the performance of DUR in detecting proteinuria. ANIMALS: Urine samples from 308 dogs and 70 cats. METHODS: Retrospective cohort study of urinalyses and UPCRs from dogs and cats collected between 2016 and 2021. RESULTS: Both canine and feline urine samples showed a positive moderate correlation between the UPCR and DUR. The correlation was not influenced by the presence of active urine sediment, glucosuria, or urine pH. In detecting canine urine samples with a UPCR >0.5, an optimal DUR of 1.4 had sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 89%, 83%, 96%, and 63%, respectively. In detecting feline urine samples with a UPCR >0.4, an optimal DUR of 2.1 had sensitivity, specificity, PPV, and NPV of 70%, 100%, 100%, and 75%, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Use of the DUR can be a relatively reliable method for identification of proteinuria. However, given its poor NPV, the DUR cannot be recommended for exclusion of proteinuric patients.

Point-of-care rapid immunoassay performed on voided urine, refrigerated up to 24 hours, accurately detects bacteriuria.

OBJECTIVE: To collect voided urine from dogs with clinical signs of lower UTI and determine the diagnostic performance of a commercially available rapid immunoassay (RIA) immediately after urine collection and after refrigeration at 4 and 24 hours. ANIMALS: 40 client-owned dogs. METHODS: Aerobic urine culture was performed on urine collected by cystocentesis. Urine samples were collected by voiding, and the RIA performed in triplicate within 30 minutes (time 0) and again in triplicate after 4 and 24 hours of refrigeration. Test precision and agreement between culture results and RIA results at each time point were determined, and factors possibly associated with false results investigated. RESULTS: 14 of 40 dogs (35%) had UTI verified by aerobic urine culture, and all had positive RIA. Three dogs had false positive RIA results. Sensitivity, specificity, positive predictive value, and negative predictive value of the RIA were 100, 88%, 82%, and 100%, respectively, and results were not different after 4 and 24 hours of refrigeration. Precision was excellent. CLINICAL RELEVANCE: This point-of-care RIA, performed on voided urine refrigerated up to 24 hours, rapidly and accurately identifies bacteriuria in dogs with lower urinary tract clinical signs, inexpensively.

The relationships between sediment findings and culture results and the presence of proteinuria in canine urine samples.

OBJECTIVES: To assess relationships between urine sediment and microbial culture findings and the presence of proteinuria in canine urine samples, and to assess the change in the percentage of proteinuric samples and urine protein-to-creatinine ratio when urine abnormalities resolve. MATERIALS AND METHODS: Canine urine samples collected via cystocentesis and submitted for culture and contemporaneous urinalysis (including urine protein-to-creatinine ratio) were retrospectively identified. Dogs receiving corticosteroids were excluded. Associations between haematuria (red blood cells>5/high-power field), pyuria (white blood cells>5/high-power field), presence of microorganisms on microscopy, active sediment, and positive culture and proteinuria (urine protein-to-creatinine ratio>0.5) were investigated. Patient characteristics were considered possible confounders. In dogs with repeat urinalysis, the associations between active sediment and positive culture resolution on proteinuria and urine protein-to-creatinine ratio were assessed. RESULTS: One hundred and ninety-two of 491 samples were proteinuric (39.1%). Age was positively associated with proteinuria. In the multivariable analysis corrected for age, active sediment was the only variable significantly associated with proteinuria (adjusted odds ratio: 2.12; 95% confidence interval: 1.44 to 3.11); however, only 49.8% of samples with active sediment were proteinuric. Neither resolution of active sediment nor positive culture were associated with reduced proportions of proteinuric samples (from 57.9% to 42.1% and from 40.0% to 25.0%, respectively) or significant reductions in urine protein-to-creatinine ratio (median change: -0.16 and -0.14, respectively). CLINICAL SIGNIFICANCE: Attributing proteinuria to urinalysis abnormalities or a positive urine culture in canine cystocentesis samples is not supported by our findings, and could result in alternative causes of proteinuria (e.g. renal proteinuria) being overlooked.

Endotoxic kidney injury in Beagle dogs assessed by serum creatinine and symmetric dimethylarginine, and urinary neutrophil gelatinase-associated lipocalin and clusterin.

Sepsis of Gram negative bacterial origin results in lipopolysaccharide-induced endotoxemia. This often leads to acute kidney injury (AKI) and its recognition remains a challenge and delays treatment. As renal damage occurs before a rise in serum creatinine is detected, new early biomarkers of kidney injury need to be explored. The aim of this study was to determine changes in serum parameters of renal function and urine biomarkers of renal injury. This was a descriptive study. Endotoxemia was induced intravenously in six anaesthetized Beagles (T1). To achieve normotension, dogs received fluids (T2), followed by a continuous infusion of noradrenaline and dexmedetomidine or 0.9% NaCl (T3). Ten minutes later, the dogs received fluids (T4) and noradrenaline and dexmedetomidine or 0.9% NaCl in a crossover manner (T5). At each timepoint, blood and urine were collected for serum creatinine, urea, symmetric dimethylarginine, urine protein/creatinine (UPC) ratio, urine neutrophil-gelatinase-associated lipocalin (U-NGAL), U-NGAL/creatinine ratio, urine clusterin (U-clusterin) and U-clusterin/creatinine ratio. Data were analyzed using a mixed-effect model taking into account time and stage of veterinary AKI (VAKI). Three of six dogs had a VAKI stage ≥1; one with anuria and elevated creatinine. Serum creatinine (P < 0.001), U-NGAL/creatinine ratio (P = 0.01) and U-clusterin/creatinine ratio increased over time (P < 0.01). The UPC ratio (mean (range) 0.68 (0.35-2.3) versus 0.39 (0.15-0.71) P < 0.01) and U-NGAL (3164 pg/mL (100-147,555) versus 100 (100-14,524), P = 0.01) were higher in VAKI stage ≥1 versus stage 0, respectively. Endotoxemia induced VAKI stage ≥1 in half of the dogs. Repeated measurement of selected parameters could detect AKI early.

Assessment of inter-observer agreement among experienced and inexperienced observers and an automated device for dipstick urinalysis in dogs and cats.

Standard visual urine dipstick analysis (UDA) is performed routinely in veterinary medicine; results can be influenced by both the operator and the method. We evaluated the agreement of results for canine and feline urine samples analyzed using a 10-patch dipstick (Multistix10SG; Siemens), both visually under double-anonymized conditions by students and a laboratory technician, and with an automated device (AD; Clinitek Status, Siemens). The mean concordance for semiquantitative urinalysis results between students and the technician and between students and the AD was fair (κ0.21-0.40) in dogs and cats; concordance was moderate between the technician and the AD (κ0.41-0.60) in dogs and good (κ0.61-0.80) in cats. For pH, the mean concordance between students and the technician and between the technician and the AD was good (ρ0.80-0.92) in dogs and cats; concordance was good between students and the AD (ρ0.80-0.92) in dogs and moderate (ρ0.59-0.79) in cats. Repeatability was higher (p < 0.001) for the technician and the AD than for a student. We found good agreement between UDA performed by an experienced operator and an AD in dogs and cats but found low reproducibility and low repeatability for urinalysis performed by an inexperienced operator.

Diagnostic cut-off values for the urinary corticoid:creatinine ratio for the diagnosis of canine Cushing's syndrome using an automated chemiluminescent assay.

BACKGROUND: Cushing's syndrome is one of the most common endocrinopathies in dogs. The preferred screening test for spontaneous Cushing's syndrome is the low-dose dexamethasone suppression test (LDDST). The diagnostic value of urinary cortisol:creatinine ratios (UCCR) is questionable. OBJECTIVES: The aim of this study was to determine diagnostic cut-off values for UCCR testing in comparison with LDDST as a clinical reference standard and to calculate the sensitivity and specificity. METHODS: Data from 2018 to 2020 were obtained retrospectively from a commercial laboratory. Both LDDST and UCCR were measured by automated chemiluminescent immunoassay (CLIA). The maximum interval between both tests was 14 days. The optimal cut-off value for UCCR testing was calculated by the Youden index. The sensitivity and specificity of these cut-off values for the UCCR test and LDDST were assessed by Bayesian latent class models (BLCMs). RESULTS: This study included 324 dogs with both UCCR test and LDDST results. The optimal UCCR cut-off value, calculated by the Youden index, was 47.4 × 10-6 . Any UCCR <40 × 10-6 was interpreted as a negative result, 40-60 × 10-6 as values in a gray zone, and >60 × 10-6 as positive. Using the cut-off of 60 × 10-6 , BLCM showed 91% (LDDST) and 86% (UCCR test) sensitivity and a specificity of 54% (LDDST) and 63% (UCCR test). CONCLUSIONS: Considering an 86% sensitivity and a 63% specificity, UCCR testing may be considered a first-line investigation to rule out Cushing's syndrome using CLIA analysis. Urine samples can be collected noninvasively at home by the owner, reducing the potential impact of stress.

Prevalence of signs of lower urinary tract disease and positive urine culture in dogs with diabetes mellitus: A retrospective study.

BACKGROUND: No recent studies have evaluated the association between clinical signs of lower urinary tract disease (LUTD) and positive urine culture in dogs with diabetes mellitus. OBJECTIVE: Determine the prevalence of subclinical bacteriuria (ie, positive urine culture without signs of LUTD) in dogs with diabetes mellitus. ANIMALS: One hundred seven dogs with diabetes mellitus were evaluated at a university veterinary hospital. METHODS: Retrospective study evaluating diabetic dogs with a single sample paired urinalysis and urine culture. Relationship between the presence of signs of LUTD, pyuria, and bacteriuria and urine culture results were compared using Fisher exact testing. RESULTS: Fifteen dogs (14%) had a positive urine culture via cystocentesis or free catch, of which 8 (53%) had pyuria, and 4 (27%) had signs of LUTD. Of the 88 dogs (82%) without signs of LUTD, 11 (13%) had a positive culture. A significant association was found between a positive urine culture and pyuria (OR infinity; 95% CI 20.34-infinity, P < .00001) and bacteriuria (OR infinity; 95% CI 164.4-infinity, P < .00001). No association was found between urine culture results and signs of LUTD (OR 1.87; 95% CI 0.59-6.85, P = .46). CONCLUSION AND CLINICAL IMPORTANCE: Subclinical bacteriuria occurred in this cohort of dogs, and our findings reinforce the recommendation that urine cultures should not be routinely performed in diabetic dogs particularly if pyuria and bacteriuria are absent.

Urinary interleukin-6 is a potentially useful diagnostic and prognostic marker of acute kidney injury in dogs.

BACKGROUND: Interleukin-6 (IL6) is a pro-inflammatory cytokine implicated in the pathophysiology of urinary tract diseases. The objective of this study was to evaluate the diagnostic and prognostic utilities of urinary IL6 (uIL6) in dogs with acute kidney injury (AKI) and other urinary tract diseases. METHODS: Eighty client-owned dogs were included and divided into four groups: AKI, chronic kidney disease (CKD), urinary tract infection and healthy controls. Urine samples were analysed for uIL6 and normalised to urinary creatinine (uIL6/uCr). RESULTS: Dogs in the AKI group had higher uIL6/uCr compared with the control and CKD groups (p < 0.001 and 0.012, respectively). Receiver operator characteristic (ROC) curve analysis of uIL6/uCr as a diagnostic marker for AKI had an area under the curve (AUC) of 0.91 (95% confidence interval [CI], 0.81-1.0) with 82% sensitivity and 90% specificity (cutoff point 4.5 pg/mg) when including the AKI and control groups. ROC analysis including AKI compared with all other groups had an AUC of 0.77 (95% CI, 0.67-0.87) for the diagnosis of AKI with sensitivity and specificity of 71% and 78%, respectively (cutoff point 10.4 pg/mg). The 30-day mortality of the AKI group was 34%, and there was no difference in uIL6/uCr between survivors and non-survivors of AKI. CONCLUSIONS: uIL6/uCr is a potentially sensitive and specific diagnostic marker for AKI in dogs.

Establishment of a reference interval for urinary albumin-to-creatinine ratio in dogs.

BACKGROUND: Albuminuria is an important marker of renal damage and can precede proteinuria; thus, it can be a useful analyte in the early diagnosis of kidney diseases. Albuminuria has also been found in dogs with hypertension, inflammatory, infectious, and neoplastic diseases. OBJECTIVES: The aim of this study was to establish a reference interval (RI) for albuminuria in dogs. METHODS: One hundred sixty-four clinically healthy dogs were enrolled in the study. Urinary albumin was determined by the immunoturbidimetric method, and albumin excretion was expressed as the urinary albumin-to-creatinine (UAC) ratio. The RI for UAC was established. RESULTS: After exclusions, 124 dogs from 32 breeds remained. The median UAC of the study population was 3.0 mg/g (range: 0-48). The RI was defined as 0-19 mg/g (with a 90% CI for the upper limit of 13-28 mg/g). No significant difference was found between male and female dogs or between different age and body weight groups. The results of Sighthounds (n = 30) and Beagle dogs (n = 23) did not differ from the other breeds. CONCLUSION: The canine RI of UAC is similar but somewhat narrower than the human RI.

Investigation of the urine cortisol to creatinine ratio for the diagnosis of hypoadrenocorticism in dogs.

OBJECTIVE: To evaluate the urine cortisol-to-creatinine ratio (UCCR) for the diagnosis of hypoadrenocorticism (HA) in dogs and to determine whether the method of urine cortisol measurement affects results. ANIMALS: 41 dogs with naturally occurring HA and 107 dogs with nonadrenal illness. PROCEDURES: Urine samples were prospectively collected from dogs undergoing testing for HA. Urine cortisol concentrations were measured at a veterinary diagnostic laboratory using either a radioimmunoassay (RIA) or a chemiluminescent immunoassay (CLIA). Receiver operating characteristic (ROC) curves were constructed to assess UCCR performance by both methods for HA diagnosis. Sensitivities, specificities, accuracies, and predictive values were calculated for various cutpoints. RESULTS: The areas under the ROC curves for UCCR diagnosis of HA were 0.99 (95% CI, 0.98 to 1.00) and 1.00 (95% CI, 1.00 to 1.00) when urine cortisol was determined by RIA and CLIA, respectively. An RIA UCCR of ≤ 2 was 97.2% sensitive, 93.6% specific, and 94.7% accurate for HA diagnosis, whereas a CLIA UCCR of ≤ 10 was 100% sensitive, specific, and accurate. An RIA UCCR > 4 and a CLIA UCCR of > 10 had negative predictive values of 100%. CLINICAL RELEVANCE: The UCCR was an accurate diagnostic test for HA in this study population, although equivocal results are possible. Case characteristics, method of cortisol measurement, and laboratory-specific cutpoints must be considered when interpreting results.

Effect of urine-specific gravity on performance of bacteriuria in predicting urine culture results.

OBJECTIVE: To determine the effect of urine-specific gravity (USG) on using microscopic evaluation of bacteriuria to predict urine culture results in dogs and cats. MATERIALS AND METHODS: We performed a retrospective medical record review of canine and feline patients that had a urinalysis and urine culture performed simultaneously. The sensitivity, specificity, positive predictive value and negative predictive value of microscopic bacteriuria for predicting urine culture results were calculated, stratified by USG. Multivariable regression was performed to test the effect of USG, pyuria, haematuria and species on the agreement between microscopic bacteriuria and culture results. RESULTS: A total of 481 dogs and 291 cats with paired urinalysis and urine culture results were included in the study. Microscopic bacteriuria had moderate sensitivity (76% in dogs, 64% in cats) and high specificity (97% in dogs, 96% in cats) for predicting urine culture bacterial growth. Samples with rod bacteria were more likely to have bacterial growth than those with cocci (OR=Infinity, 95% CI 4.8 - Infinity). As compared to isosthenuric+hyposthenuric samples (USG ≤1.012), agreement was lower in moderately concentrated (OR=0.44, 95% CI 0.19 to 0.91) samples. Absence of bacteriuria, pyuria and haematuria had a high negative predictive value for no bacterial growth (96%). CLINICAL SIGNIFICANCE: Microscopic bacteriuria has a high specificity in predicting urine culture results, regardless of USG. The finding that microscopic bacteriuria has better agreement with urine culture results in isosthenuric+hyposthenuric urine argues against reflex culture in these samples, especially if pyuria and haematuria are also absent. Urine microscopy can aid clinicians in determining the likelihood of urine culture growth.

Urinary tract infection by atypical uropathogens in dogs.

Bacterial urinary tract infection (UTI) is a common condition affecting dogs. Urine culture and antimicrobial susceptibility test, associated with the identification of underlying cause, are of primary importance in order to select a correct treatment, especially in presence of comorbidities. Two cases of immunecompromised dogs affected by urinary tract infection (UTI) have been described: the first, probably immunosuppressed due to old age, was in poor body condition, with severe odontolithiasis and periodontitis; the second was affected by chronic kidney disease in advanced stage. Urine cultures isolated two rare and atypical pathogens, Moellerella wisconsensis and Brevundimonas vesicularis, both showing sensitivity versus floroquinolones which were selected for the treatment. After a 4 weeks treatment, a second culture demonstrated the resolution of infection in both cases, in absence of clinical signs.To date neither of the two bacteria have been reported as cause of UTI in dog.

Serial serum creatinine, SDMA and urinary acute kidney injury biomarker measurements in dogs envenomated by the European adder (Vipera berus).

BACKGROUND: Acute kidney injury (AKI) is associated with high morbidity and mortality in dogs, but diagnosis may be impaired due the insensitivity of routine renal function biomarkers to detect earlier or milder forms of injury. Snake envenomation is one of several causes of AKI in dogs and humans. Dogs are commonly envenomated by the European adder (Vipera berus) between April and October each year, but few studies exist examining serial serum creatinine (sCr) and symmetric dimethylarginine (SDMA) measurements and AKI biomarkers in these dogs. Novel urinary biomarkers could improve clinical outcome by allowing earlier diagnosis of and intervention in AKI. The aim of this study was to assess the presence of AKI in dogs envenomated by V. berus at 12, 24 and 36 h after bite, as well as 14 days later, using sCr, SDMA and a panel of urinary AKI biomarkers normalised to urine creatinine (uCr), compared to a group of healthy control dogs. RESULTS: Thirty-five envenomated dogs and 35 control dogs were included. Serum creatinine did not exceed the upper reference limit at any time point in any dog after envenomation. Serum SDMA did not exceed 0.89 µmol/L in any dog. Compared to controls, urinary albumin/uCr, neutrophil gelatinase-associated lipocalin/uCr and monocyte chemotactic protein-1/uCr were significantly elevated 12 h (P <  0.0001, P <  0.0001, P = 0.01), 24 h (P <  0.001, P <  0.001, P = 0.002) and 36 h (P <  0.001, P <  0.001, P = 0.0008) after bite. Osteopontin/uCr was higher 24 and 36 h after bite (P < 0.0001), kidney injury molecule-1/uCr, interleukin-8/uCr and γ- glutamyl transferase/uCr were significantly higher 36 h after bite (P = 0.003, P = 0.0005, P = 0.001). Urinary cystatin C/uCr was not significantly different to controls at any timepoint. Biomarker/uCr ratios were not significantly different 14 days after envenomation compared to controls. CONCLUSION: Urinary biomarker/Cr ratios are indicative of mild transient, non-azotaemic AKI in dogs envenomated by V. berus.

Comparison of three types of analyzers for urine protein-to-creatinine ratios in dogs.

BACKGROUND: Quantitation of urine protein is important in dogs with chronic kidney disease. Various analyzers are used to measure urine protein-to-creatinine ratios (UPCR). OBJECTIVES: This study aimed to compare the UPCR obtained by three types of analyzers (automated wet chemistry analyzer, in-house dry chemistry analyzer, and dipstick reading device) and investigate whether the differences could affect clinical decision process. METHODS: Urine samples were collected from 115 dogs. UPCR values were obtained using three analyzers. Bland-Altman and Passing Bablok tests were used to analyze agreement between the UPCR values. Urine samples were classified as normal or proteinuria based on the UPCR values obtained by each analyzer and concordance in the classification evaluated with Cohen's kappa coefficient. RESULTS: Passing and Bablok regression showed that there were proportional as well as constant difference between UPCR values obtained by a dipstick reading device and those obtained by the other analyzers. The concordance in the classification of proteinuria was very high (κ = 0.82) between the automated wet chemistry analyzer and in-house dry chemistry analyzer, while the dipstick reading device showed moderate concordance with the automated wet chemistry analyzer (κ = 0.52) and in-house dry chemistry analyzer (κ = 0.53). CONCLUSIONS: Although the urine dipstick test is simple and a widely used point-of-care test, our results indicate that UPCR values obtained by the dipstick test are not appropriate for clinical use. Inter-instrumental variability may affect clinical decision process based on UPCR values and should be emphasized in veterinary practice.

Urinary clusterin and cystatin B as biomarkers of tubular injury in dogs following envenomation by the European adder.

Diagnosing acute kidney injury remains a challenge since the established renal biomarkers, serum creatinine (sCr) and symmetric dimethylarginine (SDMA) reflect glomerular function and not tubular injury. Sensitive tubular markers such as urinary clusterin (uClust) and cystatin B (uCysB) have been proposed to detect AKI at an earlier stage. Since envenomation by the European adder (Vipera berus berus) could serve as a spontaneous disease model of AKI we investigated these new biomarkers in affected dogs. Concentrations of uClust and uCysB as well as sCr and SDMA were analyzed retrospectively in stored samples from 26 dogs with snake envenomation and 13 healthy controls. Higher concentrations of uClust (P < 0.012) and uCysB (P < 0.001) were observed in the snake-envenomed group. Normalization of uClust and uCysB to urinary creatinine did not alter the results. No differences were observed in sCr and SDMA between the snake-envenomed group and the healthy control group. Spearman rank correlation analysis revealed a strong association of uClust with uCysB in the snake-envenomed dogs (r = 0.75 P < 0.001) but not in the healthy controls. The high percentage of snake-envenomed dogs with increased uClust and uCysB concentrations in the absence of increased sCr and SDMA suggests renal tubular injury in the affected dogs. Larger prospective case-controlled studies are warranted to evaluate the clinical utility and prognostic value of these biomarkers.

Urinary vanillylmandelic acid:creatinine ratio in dogs with pheochromocytoma.

Pheochromocytoma diagnosis in dogs is challenging because biochemical tests are not always available. In humans, urinary vanillylmandelic acid (VMA) is part of a pheochromocytoma biochemical diagnostic profile, whereas its diagnostic accuracy is currently unknown in dogs with pheochromocytoma. Prospectively, VMA was determined by HPLC and expressed as the ratio with respect to urinary creatinine (VMA:C). The diagnostic accuracy of the VMA:C ratio was evaluated by analyzing the receiver operating characteristic (ROC) curve in 10 healthy dogs, 8 dogs with pituitary-dependent hypercortisolism, 8 dogs with adrenal-dependent hypercortisolism, and 7 dogs with pheochromocytoma. The pheochromocytoma diagnosis was confirmed by histology and immunohistochemistry in all tumors. The VMA:C ratio was significantly higher in dogs with pheochromocytoma (158 [53.4 to 230.8] × 10-3) than in dogs with adrenal-dependent hypercortisolism (48.1 [24.3 to 144.9] × 10-3; P < 0.05), dogs with pituitary-dependent hypercortisolism (37.5 [32 to 47.1] × 10-3; P < 0.001), and healthy dogs (33.8 [13.3 to 87.9] × 10-3; P < 0.001). When using a VMA:C ratio >58.2 × 10-3 for pheochromocytoma diagnosis, a sensitivity of 85.7% and a specificity of 88.4% were obtained. Nevertheless, when using a cut-off ratio of 4 times the median VMA:C ratio determined in healthy dogs, there was no overlap (100% specificity). The area under the ROC curve indicated that the VMA:C ratio test could be used to discriminate between dogs with and without pheochromocytoma, what leads to the conclusion that it is useful for pheochromocytoma diagnosis in dogs.