Evaluation of modified Wright-staining of urine sediment as a method for accurate detection of bacteriuria in dogs

OBJECTIVE: To compare the findings of light microscopic evaluation of routine unstained wet-mounted preparations and air-dried, modified Wright-stained preparations of urine sediment with results of quantitative aerobic bacteriologic culture of urine. DESIGN: Masked prospective study. SAMPLE POPULATION: 459 urine samples collected by cystocentesis from 441 dogs. PROCEDURE: Urinalyses and quantitative bacteriologic cultures of urine were performed. Unstained wet-mounted preparations and air-dried, modified Wright-stained urine sediment preparations were examined by light microscopy for the presence of bacteria. RESULTS: Compared with results of quantitative bacteriologic culture, routine unstained preparations and modified Wright-stained preparations had sensitivities of 82.4% and 93.2%, specificities of 76.4% and 99.0%, positive predictive values of 40.1% and 94.5%, negative predictive values of 95.8% and 98.7%, and test efficiencies of 77.3% and 98.0%, respectively. Compared with 74 samples that yielded growth on bacteriologic culture, the routine unstained method had concordance and misclassification rates of 39.2% and 60.8%, respectively, whereas the Wright-stained method had concordance and misclassification rates of 78.4% and 21.6%, respectively. Significant associations between each of occult blood in urine, pyuria, female sex, and lower urine specific gravity with bacteriuria detected by Wright-stained sediment examination and quantitative bacteriologic culture of urine were identified. CONCLUSIONS AND CLINICAL RELEVANCE: Examination of modified Wright-stained preparations of urine sediment appeared to be a rapid, cost effective method that significantly improved the sensitivity, specificity, positive predictive value, and test efficiency of light microscopic detection of bacteriuria, compared with that of the routine unstained method.     

Comparison of the Anvajo Vet Fluidlab 1 urine sediment analyzer to manual microscopy and Idexx SediVue analysis for analysis of urine samples from cats and dogs

The Vet Fluidlab 1 (Anvajo), a new urine sediment analyzer for use in veterinary medicine, uses holographic microscopy to detect urine sediment particles in uncentrifuged urine. We compared the performance of the Fluidlab to manual microscopy and Idexx SediVue analysis for the detection of RBC, WBC, epithelial cells (EC), struvite crystals (STR), all crystals (CRY), and casts (CST) in urine samples from cats and dogs. The performance of the Fluidlab for the detection of bacteria was compared to bacterial culture. We included 624 urine samples from feline (238; 38%) and canine (386; 62%) patients; 227 samples had been submitted for bacterial culturing. The sensitivity of the Fluidlab compared to manual microscopy was 92.1% for RBC, 90.1% for WBC, 87.5% for EC, 67.6% for STR, 53.9% for CRY, and 12.5% for CST. Specificity was >97% for STR and CST, 90.0% for CRY, 78.4% for WBC, 59.4% for EC, and 55.1% for RBC. Sensitivities and specificities of the Fluidlab for analytes compared to manual microscopy were found to be similar to those obtained by the Fluidlab compared to SediVue analysis. Miscellaneous materials (e.g., lipid droplets, sperm, cell detritus) seemed to be the main reason for the high false-positive rate in RBC and EC classification by the Fluidlab. Detection of bacteria by the Fluidlab compared to bacterial culture had a sensitivity of 89.8% and a specificity of 72.3%. The performance of the Fluidlab is acceptable for the detection of WBC and bacteria; sensitivity for the detection of CRY and CST, and specificity for the detection of RBC and EC, require improvement.     

Evaluation of urine specific gravity and urine sediment as risk factors for urinary tract infections in cats

Background: It has been suggested that diseases that promote isosthenuria predispose to urinary tract infections because of a lack of the common bacteriostatic properties present in concentrated urine. Objectives: The purpose of this study was to assess the clinicopathologic risk factors for positive urine culture outcome in cats with chronic kidney disease (CKD), diabetes mellitus (DM), uncontrolled hyperthyroidism (HT), or lower urinary tract disease (LUTD). Methods: For this retrospective study, medical records of all cats in which a urinalysis and aerobic bacterial urine culture were performed between January 1995 and December 2002 were reviewed. Signalment, body weight, and clinicopathologic data were recorded. Based on the medical records, cats were diagnosed with CKD, DM, HT, or LUTD. Prevalence odds ratios and 95% confidence intervals were calculated using logistic regression. Multivariate models were created for each variable of interest while controlling for the confounding effect of disease group. Results: Six hundred fourteen cats met the criteria for inclusion in the study. Overall, positive urine cultures were identified in 16.9% of cats with CKD, 13.2% of cats with DM, 21.7% of cats with HT, and 4.9% of cats with clinical signs of LUTD. Decreasing urine specific gravity was not associated with positive urine culture when controlled for disease but pyuria, bacteriuria, and hematuria were all associated with positive urine culture outcome. Persians, females, increasing age, and decreasing body weight were all associated with positive urine culture outcome. Conclusions: Performing a urine culture sample based solely on the presence of isosthenuria does not seem warranted. Further studies are warranted to help identify host predisposing factors for urinary bacterial colonization in cats with these diseases.     

A clinical approach to multidrug-resistant urinary tract infection and subclinical bacteriuria in dogs and cats

ABSTRACT

Multidrug-resistant bacteria are increasingly isolated from the urinary tract of pets, particularly those that suffer from concurrent conditions, have been hospitalised, or were treated with antimicrobials in the recent past. Many of the multidrug-resistant bacteria encountered are resistant to all commonly used oral antibiotics. This poses both a therapeutic dilemma in the individual pet and a threat to public health. This article begins with an overview of multidrug resistance in organisms that are commonly isolated from the urinary tract of pets. This is followed by a proposed clinical approach to managing multidrug-resistant urinary bacteria, which summarises current knowledge regarding appropriate sampling and analysis, reviews the current guidelines regarding appropriate antimicrobial use and discusses treatment options that might be considered. The article highlights several shortcomings of the current knowledge to be considered when planning future clinical research and developing policies.     

Evaluation of modified Wright-staining of dried urinary sediment as a method for accurate detection of bacteriuria in cats

Background: Urinary sediment examination and quantitative urinary culture results are frequently discordant. Objectives: The aims of this study were to compare accuracy of light microscopic examination of wet‐mounted unstained (wet‐unstained) and air‐dried modified Wright‐stained (dry‐stained) sedimented preparations of urine with results of quantitative aerobic bacterial culture for detection and characterization of bacteriuria in cats. In addition, the presence of pyuria detected by urinalysis and potential risk factors were assessed. Methods: A blinded prospective study was conducted on 472 urinary samples collected from 410 cats by cystocentesis. The age and sex of each cat were recorded. Complete urinalyses were performed and included quantification of WBCs. Quantity and morphology of bacteria in each specimen were determined by light microscopic examination of wet‐unstained (performed by certified medical technologists) and dry‐stained (performed by a veterinary clinical pathologist) sedimented preparations of urine and compared with results of quantitative bacterial cultures. Results: Of 472 urinary specimens, 29 were positive for bacteriuria by culture and considered true positives and 443 were considered true negatives. Compared with these results, examination of wet‐unstained and dry‐stained urines had sensitivities of 75.9% and 82.8%, specificities of 56.7% and 98.7%, and test efficiencies of 57.8% and 97.7%, respectively. Positive likelihood ratios were 1.8 and 63.7 and negative likelihood ratios were 0.42 and 0.17 for wet‐unstained and dry‐stained examinations, respectively. Compared with 29 culture‐positive samples, the wet‐unstained method had morphologic concordance and misclassification rates of 37.9% and 62.1%, respectively, whereas the dry‐stained method had morphologic concordance and misclassification rates of 65.5% and 34.5%, respectively. Only 34% of samples with bacteriuria had pyuria. Frequency of bacteriuria was not significantly different based on age and sex of the cats, but there was a tendency for increased frequency in female cats and in cats >10 years old. Conclusions: Staining dried urinary sediment with a modified Wright‐stain significantly improved sensitivity, specificity, and test efficiency of microscopic detection and classification of bacteriuria compared with the wet‐unstained method. Pyuria should not be a criterion for determining the presence or absence of bacteriuria.     

Use of urine albumin/creatinine ratio for estimation of proteinuria in cats and dogs

The clinical utility of the urine albumin/creatinine ratio (UAC) using a simplified analyzer for estimation of proteinuria was studied in cats and dogs. Measurement results for diluted feline and canine albumin standard solutions showed linearity. Although conversion formulas (y=1.28x+1.04 and y=1.67x+10.47 for cats and dogs, respectively) were necessary, urine albumin concentrations could be determined in both animals. In cats and dogs with proteinuria, the UAC changed parallel with the urine protein/ creatinine ratio (UPC), and the Log UAC and Log UPC were significantly correlated (r=0.803 (p<0.01) in cats, r=0.801 (p<0.01) in dogs). The UAC using an UAC analyzer could be used clinically as one of the basic in-hospital laboratory tests for estimation of proteinuria in cats and dogs.     

Occurrence of occult bacteriuria in healthy cats

Knowledge of the occurrence of bacteriuria in adult, healthy cats is scarce in the scientific literature. A study was designed to investigate the occurrence of bacteriuria in healthy cats without current or previous signs of lower urinary tract disease. The study included 108 cats, 53 males (49.5%) and 55 females (50.5%). The cats ranged in age between 7 months and 18 years, with a mean age of 4.4 years and a median age of 4.0 years. Urine was obtained by cystocentesis from all the cats, and was submitted for bacteriological analyses. Urine and urine sediment was cultured on separate blood agar plates for quantification and species identification by standard procedures. Detection of ≥10(3)colony forming units (cfu) per ml urine was defined as significant bacteriuria. Significant bacteriuria exceeding 10(5) cfu/ml was detected in one sample with a combination of Enterococcus species and Staphylococcus species. There was no bacterial growth in the urine samples from 107 cats (99.1%). Results from our study indicate that the prevalence of bacteriuria in clinically healthy, adult cats is low. Also, that contamination of samples is rare when urine is collected by cystocentesis.     

Prevalence of positive urine culture in the presence of inactive urine sediment in 1049 urine samples from dogs

BackgroundUrinalysis (UA) is often used to screen for bacterial cystitis, regardless of sediment results, and followed up by quantitative urine culture (UC) for definitive diagnosis.ObjectivesDetermine prevalence of positive UCs in dogs with inactive urine sediments on routine UA.AnimalsA total of 1049 urine samples with inactive urine sediments and UCs collected from dogs presented to a veterinary specialty hospital between January 2018 and February 2020.MethodsRetrospective study of dogs with an inactive urine sediment on routine UA and follow‐up UCs. Signalment, UA findings, proteinuria, and UC results were recorded. Associations among these findings were assessed using multivariate logistic regression carried out using a backward stepwise method.ResultsOverall prevalence of positive UC was 3.4% (95% confidence interval [CI], 2.4‐4.8). Escherichia coli was the most commonly isolated bacteria. Only naturally voided samples were associated with increased prevalence of positive culture when compared to collection by cystocentesis or a non‐specified method. No statistically significant association with culture positivity was found for urine protein‐to‐creatinine ratio, urine specific gravity, urine pH, breed, age, or sex.Conclusions and Clinical ImportanceBased on the low prevalence (3.4%) of positive culture in urine samples from dogs with inactive sediment on routine UA and the relatively high cost of UC and sensitivity, cost‐benefit analysis including clinical suspicion of lower urinary tract disease should inform testing decisions, rather than routinely performing cultures on urine samples without active sediments.     

Comparison of bronchoalveolar lavage cytospins and smears in dogs and cats

Differences in the cytological interpretation of bronchoalveolar lavage fluid (BALF) after cytospin preparation (CP) or manual smearing of pelleted cells preparation (MSP) were investigated in client-owned dogs and cats with inflammatory or infectious lower respiratory disease. Bronchoalveolar lavage fluid from healthy cats was also examined. With MSP, cell lysis was more frequently observed, and cellular distribution was more heterogeneous throughout the slide. When samples from healthy and diseased animals were considered together, a significantly greater percentage of neutrophils was seen on CP than on MSP slides (P<0.002). Cytospin preparations were considered of better quality in all individual comparisons. Cytospin preparation is advised in the evaluation of BALF with low total cell count. When only MSPs are evaluated, clinicians should be aware that differential neutrophil counts may underestimate the counts found on CP slides.     

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

BackgroundQuantitation of urine protein is important in dogs with chronic kidney disease. Various analyzers are used to measure urine protein-to-creatinine ratios (UPCR).ObjectivesThis 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.MethodsUrine 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.ResultsPassing 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).ConclusionsAlthough 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.