To validate the use of a flow cytometric assay that uses 2‘,7‘-dichlorodihydrofluorescein diacetate (DCFH-DA) to measure reactive oxygen species in the erythrocytes of healthy dogs.
50 healthy adult dogs.
Erythrocytes were incubated with DCFH-DA or a vehicle control (dimethyl sulfoxide), then incubated with (stimulated) or without (unstimulated) hydrogen peroxide. The flow cytometric assay was evaluated for specificity with increasing concentrations of DCFH-DA and hydrogen peroxide, and a polynomial regression line was applied to determine optimal concentrations. For precision, samples were analyzed 5 consecutive times for determination of intra- and interassay variability. Stability of samples stored at 4°C for up to 48 hours after blood collection was determined with flow cytometric analysis. Coefficient of variation (CV) was considered acceptable at 20%. Baseline measurements were used to determine an expected range of median fluorescence intensity for unstimulated erythrocytes incubated with DCFH-DA.
Erythrocytes were successfully isolated, and stimulated samples demonstrated higher median fluorescence intensity, compared with unstimulated samples. The intra-assay CV was 11.9% and 8.9% and interassay CV was 11.9% and 9.1% for unstimulated and stimulated samples, respectively. Unstimulated samples were stable for up to 24 hours, whereas stimulated samples were stable for up to 48 hours.
CONCLUSIONS AND CLINICAL RELEVANCE
Flow cytometry for the measurement of reactive oxygen species in the erythrocytes of healthy dogs by use of DCFH-DA had acceptable specificity, precision, and stability. Flow cytometry is a promising technique for evaluating intraerythrocytic oxidative stress for healthy dogs.
To compare mineral types of naturally occurring uroliths in ferrets (Mustela putorius furo) from North America, Europe, and Asia and to identify potential risk factors associated with cystine urolithiasis in ferrets.
1,054 laboratory submission records of uroliths obtained from ferrets between January 1, 2010, and December 31, 2018.
For this cross-sectional study, the medical records databases at 4 diagnostic laboratories were searched for records of submissions of uroliths obtained from ferrets. Data collection included submission date; ferret sex, neuter status, and age; receiving laboratory and continent; and urolith mineral type. Regression analyses were performed to identify variables associated with cystine uroliths.
Of the 1,054 urolith submissions, 1,013 were from North America, with 92.6% (938/1,013; 95% CI, 90.8% to 94.1%) cystine uroliths, and 41 were from Europe and Asia, with only 26.8% (11/41; 95% CI, 15.7% to 41.9%) cystine uroliths. Median age was 2.0 years for ferrets with cystine urolithiasis versus 4.0 years for those with other types of uroliths. Submissions were more likely cystine uroliths for ferrets in North America versus Europe and Asia (adjusted OR [aOR], 59.5; 95% CI, 21.4 to 165.6), for ferrets that were younger (aOR, 0.67; 95% CI, 0.58 to 0.77), or for submissions in 2018 versus 2010 (aOR, 21.1; 95% CI, 5.1 to 87.9).
CONCLUSIONS AND CLINICAL RELEVANCE
Results indicated that the proportion of submissions that were cystine uroliths dramatically increased in North America between 2010 and 2018. There is an urgent need to determine underlying causes and mitigate cystine urolithiasis in ferrets.
Objective—To characterize and evaluate risk factors for suture-associated cystoliths in dogs and cats.
Design—Retrospective case-control study.
Animals—163 dogs and 13 cats with suture-associated cystoliths and 326 control dogs and 26 control cats with non–suture-associated cystoliths.
Procedures—Submissions to the Canadian Veterinary Urolith Centre received from 1999 to 2006 were reviewed. Case dogs and cats had cystoliths associated with visible suture or with hollow, cylindrical channels or suture knot impressions consistent with dissolved suture. Control dogs and cats had at least a single recurrent non–suture-associated cystolith submitted closest in time to the sample case. Associations among cystolith composition, recurrence times, sex, age, and breed were evaluated.
Results—Cases consisted of 92 dogs and 7 cats with visible suture and 71 dogs and 6 cats with dissolved suture. Suture-associated cystoliths represented 0.6% of canine cystoliths, 9.4% of recurrent canine cystoliths, 0.17% of feline cystoliths, and 4% of recurrent feline cystoliths. Sexually intact and neutered males were at increased odds of suture-associated cystoliths, relative to spayed female dogs. Shih Tzus, Lhasa Apsos, and Pomeranians were significantly predisposed to form suture-associated cystoliths. In dogs, compound suture-associated cystoliths were significantly more likely than other cystolith types (OR, 8.6). Dogs with suture-associated cystoliths had significantly shorter recurrence times than did control dogs.
Conclusions and Clinical Relevance—Suture remnants in the bladder have an important role in recurrent cystolithiasis in dogs. Identification of risk factors is important for avoiding recurrence of iatrogenic cystoliths.