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Objectives: To evaluate the prevalence of bacterial presence in free-catch urine samples preceded by either a standardised prepped ("clean-catch") protocol versus unprepped (non-cleaned) voiding., Materials and Methods: The study was a single-centre prospective single-blinded randomised controlled trial. Urine samples were obtained from 100 client-owned dogs presenting for routine evaluation. Dogs were randomly assigned to either the prepped group (preputial or peri-vulvar area cleaned with sterile saline before collection) or the unprepped group (no preliminary cleansing) stratified by sex. Urinalysis and urine culture (blood and MacConkey agar) were performed on all samples. Significant bacterial presence on urine culture was defined as >10 4  colony forming units (CFU)/mL., Results: There were no statistically significant associations between prepped versus unprepped collection method or sex with a urinalysis positive for bacteriuria. However, on culture, significant bacterial growth was almost five times more likely to be associated with males relative to females (odds ratio 4.59, 95% confidence interval 1.61 to 13.10). The probability of finding a positive culture was not statistically associated with prep method (odds ratio 1.43, 95% confidence interval 0.50 to 4.08)., Clinical Significance: For the majority of dogs without clinical signs of urinary tract infection, free-catch urine collection does not result in significant bacteriuria found on analysis or culture. The presence of bacteria found in free-catch samples may be secondary to sample contamination or subclinical bacteriuria. Sample contamination or subclinical bacteriuria may be more prevalent in male dogs., (© 2024 British Small Animal Veterinary Association.)

To compare the sensitivity of conjunctival swab (CS) and conventional samples (blood, spleen, liver, lymphoid and cutaneous tissue) in the diagnosis of canine visceral leishmaniasis (CVL) by polymerase chain reaction (PCR), a systematic review and meta-analysis was carried out using PubMed, Science Direct, Scopus, Web of Science, VHL/BVS (Virtual Health Library), CAPES, and Scielo databases. Articles published from 2002 to 2022 were considered and the review was updated in Jul 2023. From the total of 371 identified studies, 8 met all the eligibility criteria and were included in this review. Data from 658 CVL-positive dogs and 2541 PCR results were considered. Using a random effect model, data on the sensitivity of the test was compared between intervention (CS samples) and comparison (all the other samples) groups. Overall, the use of CS in the PCR diagnosis of CVL produced 12% higher sensitivity (p=0.013) in the test than all the other samples in combination. The animals' clinical condition did not influence (p>0.142) this overall result. However, when CS was individually compared to each of the conventional samples, the consistent result was observed (p=0.012) only in the CS versus bone marrow comparison. Given their rapid acquisition, minimal invasiveness, and lower cost relative to conventional samples, CS samples present a promising alternative for the molecular diagnosis of CVL.

Objective: To compare bacteriologic culture results for superficial swab and tissue biopsy specimens obtained from dogs with open skin wounds., Animals: 52 client-owned dogs., Procedures: For each dog, 1 wound underwent routine preparation prior to collection of 2 specimens, 1 by superficial swab (Levine) technique and 1 by tissue biopsy. Specimens were processed for bacteriologic culture. Two observers determined whether any detected difference in culture results for the 2 types of specimen would have resulted in differing treatment plans., Results: Culture results of swab and tissue biopsy specimens were identical in 11/52 (21.2%) cases. Tissue biopsy specimen and swab cultures yielded positive results for 44 (84.6%) and 40 (76.9%) wounds, respectively. With regard to mean recovery rates of bacteria from wounds with positive culture results, both the biopsy specimens and swabs yielded 3.4 bacterial species/wound. All wounds for which swab cultures yielded no growth also had negative culture results for biopsy specimens. Biopsy specimen and swab culture results were in agreement with regard to the most common bacteria cultured. In 7/52 (13%) wounds, the observers would have treated the patient differently on the basis of the results of the 2 cultures., Conclusions and Clinical Relevance: Results indicated that culture of a swab collected by the Levine technique is an appropriate noninvasive alternative to culture of a tissue biopsy specimen. A negative result obtained from culture of a swab is likely to be reliable. Disagreement between the results of swab and tissue biopsy specimen cultures is likely of low clinical importance.

Background: Quantitative bacterial culture (QBC) is the gold standard for diagnosing canine urinary tract infection. Current guidelines recommend QBC within 24 h of urine collection and that unpreserved urine is refrigerated until culture. However, temperature-controlled transport is rarely feasible, indicating a need for alternative storage during transport of urine from primary veterinary practices to the microbiology laboratory. The objective was to investigate the effect of storage temperature and boric acid sponge-preservation on quantitative bacterial culture of canine urine., Results: Significant bacteriuria was detected in 72 out of 179 samples (40%) collected from 141 dogs. Overall accuracy was 94-98% for both storage conditions and time points. Non-inferiority (15% margin) to reference quantitative bacterial culture was evident for sensitivity, specificity and predictive values for both storage methods and time points, except for the negative predictive value for 48 h boric acid preservation (NPV: 89, 95% CI [79;95]). There was no significant difference between the sensitivity and specificity for either of the time-points (p-value = 0.07-1)., Conclusions: Boric acid sponge-preservation using Uriswab™ is a useful alternative to refrigeration of urine samples during transport. Reliable quantitative bacterial culture results can be obtained from canine urine up to 48 h after collection if urine is refrigerated, and for at least 24 h if urine is stored using a boric acid-containing urine transport system., (© 2021. The Author(s).)

Objectives: To determine if lyophilized canine-specific albumin (CSA) can be stored beyond the manufacturer recommended 6 hours post-reconstitution without significant concern for bacterial growth., Design: Laboratory research., Setting: Large private teaching hospital., Interventions: Six bottles of lyophilized CSA were retrieved from a medical grade (4°C) refrigerator and were reconstituted with 0.9% NaCl to a 5% solution. Time to complete dissolution of all bottles at room temperature without agitation was recorded in minutes. A 1.5-mL sample was collected from each bottle at the following time points: time 0 (at the time of complete dissolution), and then 6, 12, and 24 hours after removal from 4°C. Each aliquot was inoculated into a blood culture tube and was immediately submitted for aerobic and anaerobic bacterial culture. A positive and negative control were cultured at the 24-hour time point. At each time point, an additional 0.5-mL aliquot from each albumin bottle was collected, and these were combined to determine the average albumin concentration (g/L, g/dL) as measured by an in-house analyzer. The albumin was stored at 4°C between sample collection., Measurements and Main Results: The reconstitution of CSA and storage at 4°C for up to 24 hours did not result in anaerobic or aerobic bacterial growth in any of the 24 cultured samples. Storage did not have a clinically significant effect on the albumin concentration of the solution. The average albumin concentration of all samples was 42 g/L (4.2 g/dL). No growth was noted in the negative control, and the positive control grew 4 strains of bacteria., Conclusions: It appears safe to use lyophilized CSA up to 24 hours post-reconstitution without significant risk of bacterial growth or change in albumin concentration., (© Veterinary Emergency and Critical Care Society 2021.)

Objective: To determine the usefulness of bacteriology swabs as a storage method of canine urine samples and the effect on quantitative bacterial culture., Materials and Methods: Two hundred fourteen canine urine samples were collected by cystocentesis. The reference aliquot was placed in a sterile tube and processed for quantitative bacterial culture within 6 hours. A bacteriology swab was then immersed in the urine for 5 seconds and returned to the charcoal Amies media container. The urine samples in the sterile tube and bacteriology swab were stored at room temperature for 48 hours and processed for quantitative bacterial culture., Results: Thirty-seven of the samples were positive on reference culture with a total of 42 bacterial isolates. Samples stored in sterile tube and bacteriology swab had identical sensitivity and specificity for detection of bacteriuria (94.7% and 100%, respectively) with very good agreement (κ = 0.92; 95% CI 0.81 to 1.00). Agreement between the bacterial species of the reference sample and the bacteriology swab was higher (κ = 0.85; 95% CI 0.71 to 0.99) than compared to the sterile tube (κ = 0.78; 95% CI 0.62 to 0.94), but the overlapping confidence intervals mean improved agreement cannot be inferred., Clinical Significance: Bacteriology swabs stored in Amies charcoal transport media should be considered an alternative method to preserve canine urine sample when immediate processing for quantitative bacterial culture is not possible. The sensitivity of culturing plain urine, stored for 48 hours in a sterile tube, for detection of bacteriuria, was higher than previously reported., (© 2020 British Small Animal Veterinary Association.)

Objective: To investigate the effects of various biological factors on total protein concentration (TPC) and serum albumin levels in canine tears., Animals Studied: 10 healthy beagles (5 female, 5 male) were used., Procedures: Experiments were conducted on separate days, collecting tears with either capillary tubes or Schirmer strips, as follows: (i) Tear collection at 3 hours intervals (from 6 am to 12 am); and (ii) Tear collection before and 20 minutes following topical histamine application (1, 10, 375 mg/mL) to induce mild, moderate, and severe conjunctivitis, respectively. TPC and serum albumin were measured with infrared spectroscopy and ELISA, respectively., Results: Tear film TPC and serum albumin ranged from 9.7-26.1 mg/mL and 6.4-1662.6 µg/mL, respectively. Protein levels did not differ significantly among time points (P ≥ .080). Median coefficient of variation (CV%) was lower with Schirmer strips compared to capillary tubes for both TPC (12% vs 15%, P = .020) and serum albumin (57% vs 78%, P = .232). TPC (P < .001), but not serum albumin was greater in male vs. female dogs. Serum albumin, but not TPC (P ≥ .099), increased significantly with each grade of conjunctivitis severity (P < .001), with no differences between collection devices (P ≥ .322); median increase was 106%, 1389%, and 2871% in eyes with mild, moderate, and severe conjunctivitis, respectively., Conclusions: There is no apparent diurnal variation in canine tear protein levels. Blood-tear barrier breakdown with conjunctivitis allows serum albumin to leak into the tear film at high concentrations. Schirmer strips compare well with capillary tubes for bioanalytical purposes in healthy and diseased eyes, and this collection method may offer improved reproducibility for protein quantification., (© 2020 American College of Veterinary Ophthalmologists.)

Piroplasm species were analyzed by molecular tools in total 31 blood samples from positive dogs, previously checked by stained slides, stored until DNA extraction between 2016 to 2018 in the laboratory Clinical Analyzes in Niterói, Rio de Janeiro. The piroplasms were identified by PCR, targeting the 18S rRNA gene and sequencing. From the total number of samples only 24 (77.4%) were positive and show adequate nucleotide sequences for interpretation with identity between 93%-100% with Babesia vogeli in compared to the sequences isolated of infected dogs from other states in Brazil deposited on GenBank. Most of dogs infected with B. vogeli had anemia (62.5%) and thrombocytopenia (95.8%). The findings of this study are compatible with previous reports in the literature and highlight B. vogeli as the most incriminated species in canine piroplasmosis in Brazil, and thrombocytopenia the hematological alteration most frequently identified in this infection. It is important to note that this is the first study involving the molecular characterization of piroplasms in the metropolitan region of Rio de Janeiro, based on PCR followed by sequencing.

The gold standard for diagnosis of colorectal masses is surgical biopsy; however, this is not always logistically or economically feasible. The authors present an alternative to established flexible and rigid endoscopic approaches when case limitations require such an approach. In seven dogs, after the identification of a mass on physical exam and computed tomographic evaluation, the colorectum was accessed using obturator-assisted prolapse to isolate discrete masses and perform shielded sampling via core needle biopsy. Histopathologic diagnosis was adequate for treatment planning in all dogs. No major complications were recorded 65-475 days after the procedure. This technique may be useful when traditional endoscopy and surgery for biopsy of colorectal masses is unavailable.

Background: Quantitative bacterial culture and susceptibility testing is the gold standard diagnostic for determining bacterial urinary tract infection. Transport of samples to external reference laboratories is common practice in veterinary medicine., Objective: To compare bacterial culture and susceptibility results from clinical urine samples when streak plate inoculation is performed immediately after sample collection versus after transport to a reference laboratory. To determine the clinical implications of discrepant culture results., Animals: One hundred and ninety-four canine and 45 feline urine samples that were submitted for urinalysis and urine culture and susceptibility testing., Methods: This was a prospective, cross-sectional study. Streak plate inoculations were performed on urine samples immediately after collection and also after transport to a reference laboratory. Samples were stored in plain sterile tubes and refrigerated up to 24 hours before transport. Culture results were compared, and discordant results were evaluated for clinical relevance. Signalment, comorbidities, lower urinary tract signs, and antimicrobial history were recorded., Results: Kappa coefficient for agreement between plating methods was 0.884. Twenty-two (71%) of 31 discrepant results were determined to have no clinical impact. Though 35% of clean midstream samples had discrepant culture results, only 8% of these had clinical impact. Conversely, 8.6% from cystocentesis were discrepant, but 41% of these had clinical impact., Conclusions and Clinical Importance: Provided urine samples are stored and transported appropriately, the immediate preplating of urine for culture and susceptibility testing is unnecessary in the majority of cases. Despite more discrepancies in plating methods for midstream samples, the minority were of clinical importance., (© 2020 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Flea identification is a significant issue because some species are considered as important vectors of several human pathogens that have emerged or re-emerged recently, such as Bartonella henselae (Rhizobiales: Bartonellaceae) and Rickettsia felis (Rickettsiales: Rickettsiaceae). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been evaluated in recent years for the identification of multicellular organisms, including arthropods. A preliminary study corroborated the usefulness of this technique for the rapid identification of fleas, creating a preliminary database containing the spectra of five species of flea. However, longterm flea preservation in ethanol did not appear to be an adequate method of storage in the context of specimen identification by MALDI-TOF MS profiling. The goal of the present work was to assess the performance of MALDI-TOF MS in the identification of seven flea species [Ctenocephalides felis (Siphonaptera: Pulicidae), Ctenocephalides canis, Pulex irritans (Siphonaptera: Pulicidae), Archaeopsylla erinacei (Siphonaptera: Pulicidae), Leptopsylla taschenbergi (Siphonaptera: Ceratophyllidae), Stenoponia tripectinata (Siphonaptera: Stenoponiidae) and Nosopsyllus fasciatus (Siphonaptera: Ceratophyllidae)] collected in the field and stored in ethanol for different periods of time. The results confirmed that MALDI-TOF MS can be used for the identification of wild fleas stored in ethanol. Furthermore, this technique was able to discriminate not only different flea genera, but also the two congeneric species C. felis and C. canis., (© 2018 The Royal Entomological Society.)

Various types of endoscopic biopsy forceps have been investigated in dogs. The Multibite (MB) are multiple-bite forceps that collect up to four tissue specimens in a single pass through the endoscope, reducing procedure time. The authors prospectively investigated its clinical utility by comparing procedure times and diagnostic quality of samples obtained with the MB to that of biopsies performed with a single-bite forceps (SB), in 21 dogs with gastrointestinal disorders. When comparing the depth, crush artefacts and diagnostic adequacy of the gastric and duodenal biopsies, there was no significant difference between the MB and SB forceps. The procedure time was significantly longer with the MB. There was no learning curve effect, and there were no reported adverse events. This study failed to demonstrate any significant clinical advantage associated with the use of the Multibite forceps over that of conventional disposable SB forceps., Competing Interests: Competing interests: None declared., (© British Veterinary Association 2018. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Canine otitis externa is a common disease. Cytological evaluation of otic exudate is a useful diagnostic test to direct and monitor treatment for otitis externa. One method of collecting ear cytological specimens utilizes a cotton-tipped swab inserted into the vertical ear canal. A proposed alternative method is to aspirate exudate from the deep horizontal canal using a rubber tube., Hypothesis/objectives: To compare cytological findings between two different collection techniques by evaluating the numbers of bacteria, Malassezia yeast and inflammatory cells., Animals: Thirty client-owned dogs with otitis externa., Methods and Materials: A prospective, randomized, blinded comparison study. Ear canals from each dog were sampled using cotton swab and rubber tube. Ear cytological preparations were evaluated independently by two investigators for polymorphonuclear leucocytes (PMN), monocytes/lymphocytes, macrophages, yeast, intracellular (IC) cocci, extracellular (EC) cocci, IC bacilli and EC bacilli., Results: A paired Student's t-test was used to compare the two techniques. The inter-investigator reliability for PMN, EC bacilli and yeast was good, and for EC cocci was moderate. There were significantly higher numbers of PMNs obtained by the tube method (P = 0.0024) than by the cotton swab method. There were no statistically significant differences between cotton swab and rubber tube methods for monocytes/lymphocytes (P = 0.7780), macrophages (P = 0.1751), EC cocci (P = 0.1262), EC bacilli (P = 0.1162), yeast (P = 0.5371), IC cocci (P = 0.6606) or IC bacilli (P = 0.6761). The technique was well tolerated., Conclusion: An alternative ear cytological collection technique was identified which enables sampling of the deep horizontal canal., (© 2018 ESVD and ACVD.)

OBJECTIVE To examine the effect of 24 hours of refrigeration on urine samples collected from dogs with signs of urinary tract infection (UTI). DESIGN Prospective cross-sectional study. ANIMALS 104 dogs with signs consistent with UTI that had a urine sample collected via cystocentesis as part of their diagnostic workup. PROCEDURES A 1-mL aliquot of each urine sample was refrigerated at 5°C for 24 hours in a plain glass tube, then processed for quantitative bacterial culture (QBC). A 0.5-mL aliquot was added to 3 mL of tryptic soy broth (TSB) and refrigerated at 5°C for 24 hours, then processed for QBC. The remaining portion was immediately processed for QBC, with results reported as numbers of bacterial colony-forming units (CFUs). Sensitivity of the QBC for detection of bacteria (and therefore UTI) was determined for sample refrigeration in the 2 conditions, compared with immediate processing (reference standard). RESULTS Bacterial growth was identified in 35.6% (n = 37), 33.7% (35), and 31.7% (33) of the immediately processed, refrigerated, and refrigerated-in-TSB urine samples, respectively. Sample refrigeration without TSB resulted in no significant difference in CFU counts relative to immediate processing; however, the sensitivity of this method was 95% (35/37). Sample refrigeration with TSB resulted in significantly lower CFU counts, and sensitivity was only 89% (33/37). CONCLUSIONS AND CLINICAL RELEVANCE Canine urine samples collected for bacterial culture should be immediately submitted for testing. Although CFU counts for refrigerated and immediately processed samples were statistically similar in this study, sample refrigeration in enrichment broth resulted in imperfect sensitivity for UTI detection and is not recommended.

Dry-mount fecal cytology (FC) is a component of the diagnostic evaluation of gastrointestinal diseases. There is limited information on the possible effect of the sampling method on the cytologic findings of healthy dogs or dogs admitted with diarrhea. We aimed to: (1) establish sampling method-specific expected values of selected cytologic parameters (isolated or clustered epithelial cells, neutrophils, lymphocytes, macrophages, spore-forming rods) in clinically healthy dogs; (2) investigate if the detection of cytologic abnormalities differs among methods in dogs admitted with diarrhea; and (3) investigate if there is any association between FC abnormalities and the anatomic origin (small- or large-bowel diarrhea) or the chronicity of diarrhea. Sampling with digital examination (DE), rectal scraping (RS), and rectal lavage (RL) was prospectively assessed in 37 healthy and 34 diarrheic dogs. The median numbers of isolated ( p = 0.000) or clustered ( p = 0.002) epithelial cells, and of lymphocytes ( p = 0.000), differed among the 3 methods in healthy dogs. In the diarrheic dogs, the RL method was the least sensitive in detecting neutrophils, and isolated or clustered epithelial cells. Cytologic abnormalities were not associated with the origin or the chronicity of diarrhea. Sampling methods differed in their sensitivity to detect abnormalities in FC; DE or RS may be of higher sensitivity compared to RL. Anatomic origin or chronicity of diarrhea do not seem to affect the detection of cytologic abnormalities.

Giardia infection is a common clinical problem in humans and pets. The diagnosis of giardiasis is challenging as hosts intermittently excrete protozoan cysts in their feces. In the present study, we comparatively evaluated two methods of serial fecal sampling in humans, dogs, and cats from Rio de Janeiro, Brazil. The Faust et al. technique was used to examine fecal specimens collected in triplicate from 133 patients (52 humans, 60 dogs, and 21 cats). Specimens from 74 patients were received from the group assigned to carry out sampling on consecutive days - 34 humans, 35 dogs, and 5 cats, and specimens from 59 patients were received from the group assigned to carry out sampling on non-consecutive, separate days - 18 human beings, 25 dogs, and 16 cats. G. duodenalis cysts were found in stools of 30 individuals. Multiple stool sampling resulted in an increase in the number of samples that were positive for Giardia in both groups. The authors therefore conclude that multiple stool sampling increases the sensitivity of the Faust et al . technique to detect G. duodenalis cysts in samples from humans, cats and dogs.

Cutaneous cytologic sampling techniques are used to detect bacteria, yeast, and inflammatory cells for diagnosis and therapeutic monitoring. Studies have examined slide evaluation techniques, ear swab cytology staining methods, and observer variations; few studies compare common clinical sampling techniques. The primary aim of this study was to measure detection of microorganisms and neutrophils by impression smear compared to acetate tape preparation; comparison of agreement between two acetate tape staining methods was a secondary aim. Thirty lesions consistent with superficial pyoderma were sampled via impression smear and acetate tape preparation. Acetate tape preparations were either stained with modified Romanowksy stain solutions two and three or solution three alone. Impression smears were stained in the standard manner. Bacteria, yeast, and neutrophils were evaluated using a semi-quantitative scale [0-4]. Quantities were aggregated and compared using Cohen's kappa to measure agreement between methods. When impression smears were compared to acetate tape, the lowest agreement occurred for neutrophils, with impression smears detecting more neutrophils. Comparison of acetate tape staining methods had the highest agreement for yeast detection. Sampling technique and staining method did not differ for detection of bacteria. Impression smears detected more neutrophils, and yeast detection appeared equivalent for acetate tape staining methods.

Strongyloides stercoralis is a gastrointestinal parasitic nematode with a life cycle that includes free-living and parasitic forms. For both clinical (diagnostic) and environmental evaluation, it is important that we can detect Strongyloides spp. in both human and non-human fecal samples. Real-time PCR is the most feasible method for detecting the parasite in both clinical and environmental samples that have been preserved. However, one of the biggest challenges with PCR detection is DNA degradation during the postage time from rural and remote areas to the laboratory. This study included a laboratory assessment and field validation of DESS (dimethyl sulfoxide, disodium EDTA, and saturated NaCl) preservation of Strongyloides spp. DNA in fecal samples. The laboratory study investigated the capacity of 1:1 and 1:3 sample to DESS ratios to preserve Strongyloides ratti in spike canine feces. It was found that both ratios of DESS significantly prevented DNA degradation compared to the untreated sample. This method was then validated by applying it to the field-collected canine feces and detecting Strongyloides DNA using PCR. A total of 37 canine feces samples were collected and preserved in the 1:3 ratio (sample: DESS) and of these, 17 were positive for Strongyloides spp. The study shows that both 1:1 and 1:3 sample to DESS ratios were able to preserve the Strongyloides spp. DNA in canine feces samples stored at room temperature for up to 56 days. This DESS preservation method presents the most applicable and feasible method for the Strongyloides DNA preservation in field-collected feces., Competing Interests: The authors declare no conflicts of interest.

Histopathologic evaluation of liver biopsy specimens yields information that is not otherwise obtainable and is frequently essential for diagnosing hepatic disease. Percutaneous needle biopsy, laparoscopic biopsy, and surgical biopsy each have their own set of advantages and disadvantages. Care should be taken to ensure an adequate amount of tissue is collected for meaningful histologic evaluation. Because sampling error is a limitation of hepatic biopsy, multiple liver lobes should be biopsied. This article discusses the indications for liver biopsy, associated risks, advantages and disadvantages of different biopsy techniques, and strategies to get the most useful information possible out of this process., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Assessment of health status and the course of treatment of patients are often determined by results obtained from analysis of blood parameters. Errors in results can occur and cause inappropriate interpretation of laboratory data and, therefore, disease states. Electrolytes and minerals are particularly prone to spurious results; therefore, it is critical that factors influencing inappropriate resulting be recognized, and steps taken to minimize them., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background: Up to 25% of elderly humans have proteinuria, often associated with underlying lesions. Data concerning the presence of proteinuria in elderly dogs is scarce., Objectives: To describe the presence and persistence of proteinuria and to compare urinary protein : creatinine ratio (UPC) between free catch and cystocentesis urine samples in apparently healthy elderly dogs., Animals: Hundred apparently healthy elderly dogs., Methods: Prospective study. Owners of 100 elderly dogs were asked to collect 2 free catch urine samples. Dogs were considered healthy based on owner's perception and an age chart, based on ideal bodyweight, was used to define dogs as senior or geriatric. UPC of urine collected by free catch and cystocentesis were compared. Overt proteinuria and borderline proteinuria were defined as UPC >0.5 and between 0.2 and 0.5, respectively, if examination of sediment did not explain proteinuria. Proteinuria was considered persistent if present at both sampling times., Results: At baseline, 71 owners succeeded in collecting urine. Eleven percent of dogs had overt proteinuria, 14% were borderline proteinuric, and 75% nonproteinuric. Thirty-seven repeated urine samples, with a median time interval of 31 days (range 10-90), were available. Nineteen percent of dogs had a persistently increased UPC (>0.2), with persistent overt proteinuria present in 8%. A strong correlation (ρ = 0.88) was found between UPC of urine collected by free catch and cystocentesis., Conclusions and Clinical Importance: As 19% of study dogs had persistent proteinuria, our findings emphasize that measurement of proteinuria should be part of geriatric health screening. For UPC in dogs, free catch urine provides a good alternative to cystocentesis., (Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

The use of voided urine specimens for bacteriological culture in dogs is discouraged because contamination from external genitalia could lead to misinterpretation of laboratory results. Quantitative culturing and defining significant bacteriuria could increase the usefulness of voided specimens. However, limited evidence exists for the cut-offs currently recommended. The aim of this study was to evaluate the accuracy of current veterinary cut-off values for significant bacteriuria in voided canine urine. A secondary aim was to investigate if accuracy improved when applying qualitative criteria used in humans. Paired urine specimens were collected by both cystocentesis and voiding, and quantitative bacteriological cultures were performed within the same day. Cystocentesis was used as the reference standard with a cut-off for significant bacteriuria of ≥1000 colony forming units (CFU)/mL. Voided specimens were compared to cystocentesis using: (1) the veterinary cut-off of ≥100,000 CFU/mL; and (2) various cut-offs depending on qualitative criteria (sex, clinical signs and complicating factors), adapted from human guidelines. Ninety-four dogs with suspected urinary tract infection (UTI) were included for analysis. The veterinary cut-off yielded an accuracy of 94% with a sensitivity and specificity of 94% (95% confidence intervals [CI] 0.81, 0.99) and 94% (95% CI 0.86, 0.98), respectively. Applying the human guidelines did not improve overall accuracy (89%), and yielded a sensitivity and specificity of 97% (95% CI 0.86, 1.00) and 86% (95% CI 0.77, 0.92), respectively. The veterinary cut-off value of ≥100,000 CFU/mL for voided urine is appropriate for determining significant bacteriuria in the majority of dogs with suspected UTI if specimens are refrigerated and cultured on the day of collection., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: Limited evidence exists in the literature regarding whether a specific mount is preferable to use for processing endoscopically obtained gastrointestinal biopsy specimens., Hypothesis/objectives: To compare 3 methods of handling endoscopically obtained gastrointestinal biopsy specimens from collection to laboratory processing and to determine if any technique produced superior results., Animals: Twenty-three dogs and cats presented for gastrointestinal signs., Methods: Prospective study of dogs and cats presented with gastrointestinal signs to a veterinary teaching referral hospital which underwent upper gastrointestinal endoscopy. Biopsy specimens were taken from the stomach and duodenum and submitted to the laboratory using 3 techniques: mounted on a cucumber slice, mounted on a moisturized synthetic foam sponge, and floating free in formalin. The techniques were compared with regard to the specimens' width, orientation, presence of artifacts, and pathologist's confidence in diagnosis., Results: Twenty-three patients were included, with a total of 528 biopsies collected. Specimens on cucumber slice and on sponge were significantly wider (P < .001 and P = .001, respectively) compared to those floating free in formalin (mean width of 3.81 versus 3.31 and 2.52 mm, respectively). However, specimens on synthetic sponge had significantly fewer artifacts compared to those on cucumber slice (P = .05) and those floating free in formalin (P = .02). Confidence in the diagnosis also was superior with the sponge technique over floating free specimens (P = .002)., Conclusions and Clinical Importance: The use of mounted gastrointestinal biopsy specimens was superior over the use of specimens floating free in formalin. This technique improved the quality of the specimens and the pathologist's confidence in their histopathologic interpretation., (Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Objective: To assess the effect of various tissue apposition techniques on specimens after formalin fixation, and to assess canine cadaveric composite skin-muscle-fascia specimen changes after excision without and with addition of a tumor model., Study Design: Canine cadaveric study; unbalanced paired design., Animals: Fresh canine cadavers (n = 30) METHODS: Experiment 1: Paired 7 cm diameter, circular skin-muscle-fascia specimens were harvested bilaterally from three sites (thorax, abdomen, and thigh) on 10 cadavers. For each pair, the skin-to-fascia was sutured with a circumferential continuous pattern in one specimen and left unsutured in the other during excision. Specimen length and width were measured post-excision and post-fixation. Specimen rotation, translation, and distortion was measured post-fixation. Experiment 2: A subcutaneous tumor model was implanted at 2 sites (neck, thorax) on 20 cadavers and specimens were then excised with 2 cm lateral margins and a deep fascial plane. Three suturing techniques of the skin-to-fascia at both sites were performed during excision: unsutured versus circumferential continuous suture (10 pairs) or circumferential versus 4 interrupted quadrant sutures (10 pairs). Lateral margins were compared post-excision and post-fixation., Results: Experiment 1: There was significant reduction in the length and width of the fascia and skin post-excision, but no further reduction after fixation. The circumferential suture decreased rotation and translation of the tissue layers but did not significantly affect specimen shrinkage or distortion compared with the unsutured specimen. Experiment 2: All 4 lateral margins decreased significantly post-excision and post-fixation compared with the planned pre-excision margins. No difference was found between any of the 4 lateral margins for any skin-to-fascia suture technique., Conclusion: Canine skin-muscle-fascia specimens significantly decrease in size from gross pre-excision dimensions after removal. Suturing the specimen during surgery before excision will significantly improve alignment of tissue planes without creating distortion of the sample, but may alter the length, width, and lateral margins of the specimen., (© Copyright 2016 by The American College of Veterinary Surgeons.)

Traditionally, histological investigations of the epileptic brain are required to identify epileptogenic brain lesions, to evaluate the impact of seizure activity, to search for mechanisms of drug-resistance and to look for comorbidities. For many instances, however, neuropathological studies fail to add substantial data on patients with complete clinical work-up. This may be due to sparse training in epilepsy pathology and or due to lack of neuropathological guidelines for companion animals.The protocols introduced herein shall facilitate systematic sampling and processing of epileptic brains and therefore increase the efficacy, reliability and reproducibility of morphological studies in animals suffering from seizures.Brain dissection protocols of two neuropathological centres with research focus in epilepsy have been optimised with regards to their diagnostic yield and accuracy, their practicability and their feasibility concerning clinical research requirements.The recommended guidelines allow for easy, standardised and ubiquitous collection of brain regions, relevant for seizure generation. Tissues harvested the prescribed way will increase the diagnostic efficacy and provide reliable material for scientific investigations.

Objective: To describe a surgical technique for laparoscopic medial iliac lymph node (MILN) extirpation, and to describe the quality of biopsy specimens obtained., Design: Experimental study., Animals: Purpose-bred male hound-mix research dogs (n = 8)., Methods: Dogs were randomized to groups of left or right-sided laparoscopic surgical approaches. Three transperitoneal portals were established with the dogs in lateral recumbency, and ipsilateral MILN dissection was achieved under CO2 pneumoperitoneum using a vessel-sealing device., Results: MILN ipsilateral to the approach were successfully identified and removed laparoscopically in 8 dogs. Observed complications included mild to moderate hemorrhage that was controlled laparoscopically in 4 dogs, and tearing of the MILN capsule during retraction and dissection in 3 dogs. No other major complications occurred and all dogs recovered uneventfully. Areas of either minor peripheral (9/9) or central (4/9) pinch artifact affected a median percentage of 5% of surface area (range, 5-30%) of bisected lymph nodes., Conclusions: Laparoscopic MILN extirpation is feasible in dogs with normal MILN and may serve as a minimally invasive approach for excisional biopsy in the diagnostic staging of canine onocologic patients with normal-sized MILN. This lateral laparoscopic approach allows dissection of the ipsilateral MILN but precludes removal of the contralateral MILN. Minimal handling of the lymph node during dissection and removal is required to reduce the risk of capsular tear, or introduction of possible histologic artifact by tissue crush that may impact diagnosis., (© Copyright 2014 by The American College of Veterinary Surgeons.)

Background: C-reactive protein (CRP) is a major acute phase protein in both people and dogs., Objective: The objective of this study was to evaluate the precision and accuracy of 3 different point-of-care tests (POCT) for canine CRP (cCRP) in comparison to a standard ELISA test, and to assess storage stability., Material and Methods: Blood samples were collected from 125 dogs (23 healthy and 102 diseased). Serum cCRP measurements were performed using the TECOmedical AG, the EUROLyser Diagnostica, and the LifeAssays POCT. The TECOmedical AG ELISA validated for canine serum was used as a reference method. Statistical analyses included descriptive statistics, inter assay variance for each POCT, comparison of results from each POCT with the ELISA using Spearman's correlation and Bland-Altman plots, and a t-test to evaluate sample stability., Results: ELISA cCRP values ranged from 0.1-4.7 mg/L (median 1.3) in healthy dogs and from 0-282 mg/L (median 17.3) in diseased dogs. Spearman's correlation coefficient between ELISA and the respective POCT measurements for cCRP concentration was 0.89 for TECOmedical, 0.85 for EUROLyser, and 0.97 for LifeAssays. Bias calculated from Bland-Altman difference plots was 27.6% for TECOmedical, -14.2% for EUROLyser, and -15.7% for LifeAssays. Inter assay reliability was > 0.9 for all POCT. Total observed error of the EUROLyser was 28.2% and therefore met the acceptable total error of 29.6%., Conclusions: All POCTs were able to measure cCRP, but precision, accuracy, and correlation coefficients varied among the 3 systems. Therefore, serial measurements for monitoring of cCRP in dogs should always be performed using the same POCT system., (© 2015 American Society for Veterinary Clinical Pathology.)

Background: Previously, we evaluated a minimally invasive epidermal lipid sampling method called skin scrub, which achieved reproducible and comparable results to skin scraping. The present study aimed at investigating regional variations in canine epidermal lipid composition using the skin scrub technique and its suitability for collecting skin lipids in dogs suffering from certain skin diseases. Eight different body sites (5 highly and 3 lowly predisposed for atopic lesions) were sampled by skin scrub in 8 control dogs with normal skin. Additionally, lesional and non-lesional skin was sampled from 12 atopic dogs and 4 dogs with other skin diseases by skin scrub. Lipid fractions were separated by high performance thin layer chromatography and analysed densitometrically., Results: No significant differences in total lipid content were found among the body sites tested in the control dogs. However, the pinna, lip and caudal back contained significantly lower concentrations of ceramides, whereas the palmar metacarpus and the axillary region contained significantly higher amounts of ceramides and cholesterol than most other body sites. The amount of total lipids and ceramides including all ceramide classes were significantly lower in both lesional and non-lesional skin of atopic dogs compared to normal skin, with the reduction being more pronounced in lesional skin. The sampling by skin scrub was relatively painless and caused only slight erythema at the sampled areas but no oedema. Histological examinations of skin biopsies at 2 skin scrubbed areas revealed a potential lipid extraction from the transition zone between stratum corneum and granulosum., Conclusions: The present study revealed regional variations in the epidermal lipid and ceramide composition in dogs without skin abnormalities but no connection between lipid composition and predilection sites for canine atopic dermatitis lesions. The skin scrub technique proved to be a practicable sampling method for canine epidermal lipids, revealed satisfying results regarding alterations of skin lipid composition in canine atopic dermatitis and might be suitable for epidermal lipid investigations of further canine skin diseases. Although the ceramide composition should be unaffected by the deeper lipid sampling of skin scrub compared to other sampling methods, further studies are required to determine methodological differences.

Background: Limited availability of diagnostic cytopathologic material may preclude additional diagnostic techniques. Tissue transfer allows for preparation of additional slides from a single original slide. Information pertaining to the application of the tissue transfer technique in veterinary cytopathology is lacking., Objectives: The objectives were to evaluate the application of the tissue transfer technique on Quick Dip-stained veterinary cytologic smears and to assess if a selection of histochemical and immunocytochemical stains, and PCR analyses could be performed on transferred material., Methods: Archived Quick Dip-stained canine lymph node aspirate smears from previously diagnosed lymphoma cases were utilized to validate and optimize the tissue transfer technique. In this technique, diagnostic material is lifted from the original stained slide, is divided and transferred to multiple new slides. Histochemical stains such as Gram, periodic acid Schiff, Congo red, and Ziehl-Neelson, immunohistochemistry for CD3 and PAX5, and PCR for cryptococcal and mycobacterial organisms were selectively performed on transferred material., Results: The tissue transfer technique was simple, and transferred Quick Dip-stained material retained cellular morphology. Histochemical and immunohistochemical stains, and PCR analysis yielded reliable results when performed on the additional smears produced by this technique., Conclusions: The tissue transfer technique was simple and easy to perform on previously Quick Dip-stained cytology smears. Cellular detail was preserved and multiple additional ancillary diagnostic techniques were facilitated, such as histochemical and immunohistochemical stains, and PCR analysis., (© 2014 American Society for Veterinary Clinical Pathology and European Society for Veterinary Clinical Pathology.)

Background: Changes in canine hematology measurements may occur when analyses are delayed due to shipment of specimens to a laboratory., Objective: The purpose of this study was to report changes in hematologic variables in healthy and diseased canine blood measured with a Sysmex XT-2000iV during storage at room temperature for 24 and 48 hours., Methods: EDTA-K3 blood samples from 42 healthy and diseased dogs were measured on a Sysmex XT-2000iV analyzer within one hour of sampling, and after storage for 24 and 48 hours at room temperature in the dark., Results: Storage caused little or no change in RBC count, HGB concentration and MCH, while there was a moderate increase in HCT, MCV and reticulocytes count, and a moderate decrease in MCHC. Decreased platelet counts by impedance (PLT-I) and optical (PLT-O) measurements were associated with increased mean platelet volume (MPV), platelet-large cell ratio (P-LCR) and platelet distribution width (PDW), including a right shift in the platelet histogram and a dispersion of the platelet dot plot on the scattergram. The total and differential WBC count remained stable except for decreased monocyte counts. In the scatterplots, monocytes shifted into the lymphocyte population after 24 hours, and neutrophil population shifted to the right appearing in the eosinophil gate at 48 hours of storage. The disease status had only a small effect on storage-induced changes, and observed changes had no consequences for clinical decisions., Conclusions: Blood storage at room temperature was accompanied by moderate variations in some hematologic variables, awareness of which helps in avoiding misinterpretations., (© 2014 American Society for Veterinary Clinical Pathology and European Society for Veterinary Clinical Pathology.)

Cerebrospinal fluid (CSF) is a potential source for disease-specific biomarkers that may assist in the staging and determining the prognosis of neurodegenerative conditions in animals. However, the validity of such putative biomarkers may be influenced by pre-analytical variables, including the procedures adopted to collect and store the CSF. This study assessed the effect of three handling practices on the stability of a panel of CSF proteins: clusterin (also known as apolipoprotein J), haptoglobin, cystatin C, and transthyretin (TTR). The three handling procedures for canine CSF were mimicked in the laboratory as follows: (1) storage in a refrigerator overnight (4 °C for 18 h); (2) carrying a sample in the pocket of a clinician (37 °C for 4h); and (3) mailing a sample to a remote laboratory for analysis (room temp for 48 h). The impact of these three scenarios on the concentrations of the selected proteins was assessed using Western blotting and compared to an aliquot of CSF that had been kept frozen. The level of clusterin was significantly reduced following 48 h at room temperature (P<0.05), while the concentration of the dimeric form of TTR increased following this handling procedure and also when held at 37 °C for 4h. A reducing agent prevented this increase at 37 °C. In conclusion, exposing CSF samples to various environmental conditions can significantly alter their protein content, a factor that must be considered in studies assessing potential biomarkers in canine CSF., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

For cases requiring both a bone marrow aspirate and core biopsy, using the same needle and site (i.e., a combined technique) can decrease time, expense, and discomfort compared with the independent (direct) collection of each specimen. The benefits of the combined approach should not be achieved at the expense of specimen quality. In this study, core bone marrow samples obtained from the proximal humerus of 26 dogs by a combined technique immediately posteuthanasia were compared with core samples obtained directly from the opposite humerus. Both core samples from each dog were compared for relative overall quality. Biopsies were unsuccessful in four dogs using the combined technique and in one dog by the direct technique. Marrow length was shorter and hemorrhage artifact was more common using the combined technique. There were no differences in cellularity, megakaryocyte count, the myeloid/erythroid ratio (M/E ratio), iron stores, or diagnostic quality. Direct core biopsy remains the ideal technique; however, the degree of compromise was small in the samples obtained from the combined technique. For clinical patients, the advantages of the combined technique must continue to be weighed against potential loss of diagnostic sensitivity.

Adapter-modified Ussing chambers have been used for assessment of endoscopically obtained intestinal biopsies in humans. The aim of this study was to evaluate the feasibility of an adapter-modified Ussing chamber for assessment of intestinal transport physiology in endoscopically-obtained colonic biopsies from cats and dogs. Fifteen colonic biopsies from four cats and 13 colonic biopsies from four dogs were transferred into a modified Ussing chamber and sequentially exposed to several compounds. Baseline mean±SD conductance was measured. Changes of short circuit current (ΔIsc) were observed after exposure to glucose (number of feline biopsies that responded=0/number of canine biopsies that responded=4), phloridzin (n=0/n=7), histamine (n=5/n=12), serotonin (n=7/n=12), prostaglandin (n=5/n=7), forskolin (n=7/n=7), and ouabain (n=9/n=7). The adapter-modified Ussing chamber studied here enables investigation of transport physiology of endoscopically-obtained colonic biopsies from companion animals. However, we observed a large variability of results, suggesting that clinical use of this method is limited., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Objective: To determine whether preanalytic and analytic factors affect evaluation of the urinary protein-to-creatinine (UPC) ratio in dogs., Sample: 50 canine urine samples., Procedures: The UPC ratio was measured to assess the intra-assay imprecision (20 measurements within a single session), the influence of predilution (1:10, 1:20, and 1:100) for urine creatinine concentration measurement, and the effect of storage at room temperature (approx 20°C), 4°C, and -20°C., Results: The coefficient of variation at room temperature determined with the 1:20 predilution was < 10.0%, with the highest coefficients of variation found in samples with a low protein concentration or low urine specific gravity. This variability could result in misclassification of samples with UPC ratios close to the thresholds defined by the International Renal Interest Society to classify dogs as nonproteinuric (0.2), borderline proteinuric (0.21 to 0.50), or proteinuric (> 0.51). A proportional bias was found in samples prediluted 1:10, compared with samples prediluted 1:20 or 1:100. At room temperature, the UPC ratio did not significantly increase after 2 and 4 hours. After 12 hours at room temperature and at 4°C, the UPC ratio significantly increased. The UPC ratio did not significantly change during 3 months of storage at -20°C., Conclusions and Clinical Relevance: The intra-assay precision of the UPC ratio was sufficiently low to avoid misclassification of samples, except for values close to 0.2 or 0.5. The optimal predilution ratio for urine creatinine concentration measurement was 1:20. A 1:100 predilution is recommended in samples with a urine specific gravity > 1.030. The UPC ratio must be measured as soon as samples are collected. Alternatively, samples should be immediately frozen to increase their stability and minimize the risk of misclassification of proteinuria.

Objective: To identify the optimal method of submission of canine and feline urine for bacterial culture., Methods: Cystocentesis samples from 250 animals (200 dogs, 50 cats) suspected of having urinary tract infections were collected. The reference aliquot, without preservative, was processed on site within 2 hours. Two further aliquots (one without preservative, one with boric acid) were stored at room temperature for up to 7 hours and then posted by guaranteed next day delivery to a commercial laboratory for analysis., Results: Forty-seven of the samples were positive on culture in the reference test. There was no significant difference between reference test results and those of samples posted without preservative (P=0·39), but samples posted in boric acid were significantly less likely to give a positive result (P=0·01). Samples posted without preservative had a sensitivity of 82% and a specificity of 98%; for boric acid, sensitivity was 73% and specificity 99%., Clinical Significance: Postal urine samples should be submitted to the laboratory in a plain sterile tube., (© 2011 British Small Animal Veterinary Association.)

Intranasal tumors of dogs and cats pose a diagnostic and therapeutic challenge for small animal practitioners. Multiple nasal biopsy techniques have been described in the past. This report describes a simplified flushing technique to biopsy and debulk nasal tumors, which often also results in immediate clinical relief for the patient. Based on the results of this retrospective study, the authors recommend high-pressure saline hydropulsion as a minimally invasive diagnostic, and potentially therapeutic, technique for nasal tumors in dogs and cats.

There is little information in veterinary literature regarding the diagnostic accuracy of aspirate cytology for the diagnosis of canine osteosarcoma (OSA). The authors compared the diagnostic accuracy of a novel method of cytologic collection, termed core aspirate cytology (CA), with fine needle aspiration (FNA) and histopathology in 27 dogs with lytic and/or proliferative bone lesions. Alkaline phosphatase (ALP) staining was performed to confirm the diagnosis of OSA cytologically. OSA was accurately diagnosed in 85% and 95% of FNA and CA, respectively. ALP staining was 100% sensitive for the diagnosis of OSA. CA using a bone marrow biopsy needle allowed for penetration of cortical bone and aspirate cytology with a larger bore needle than FNA; however, there was no significant difference in diagnostic accuracy between techniques. Aspirate cytology with ALP staining was a safe, accurate, and minimally invasive diagnostic test for the evaluation of suspected OSA lesions in dogs.

Fine-needle aspirates and impression smears of cutaneous and subcutaneous lesions and lymph nodes are the most commonly submitted cytology samples from veterinary patients. Diagnostic cytology samples of these lesions are easily collected in patients without anesthesia or analgesia. Cytology can yield immediate results and may prevent the need for additional tests that use more invasive methods of sample collection. This article offers a brief review of how to collect and submit cytology samples and describes cytologic lesions that often are diagnosed in dogs and cats. When applicable, differences between disease progression in dogs and cats are described., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Urinary markers for renal dysfunction are gaining interest, but effects of sampling method, storage conditions, and urinary tract inflammation or infection on these markers are unclear. Therefore, the objectives of the current study were to determine the difference in urinary albumin (uALB), urinary retinol-binding protein (uRBP), and urinary N-acetyl-β-D-glucosaminidase (uNAG) concentrations in cystocentesis and voided samples and to investigate concentration changes after storage at -20°C and at -80°C. Effects of a protease inhibitor were also assessed in samples stored at -80°C for 12 months. In a pilot experiment, influence of in vitro hematuria, pyuria, and bacteriuria on the urinary markers was evaluated. A mixed model was used to calculate mean differences and 95% confidence intervals. Urinary ALB, uNAG, and uRBP concentrations were similar in voided and cystocentesis samples. After storage for 4 months at -20°C, uALB concentration was not affected, and uRBP concentration showed a mild and clinically irrelevant decrease, whereas uNAG activity was significantly lower compared with fresh samples. After storage for 12 months at -80°C, uALB and uRBP concentrations did not differ from fresh samples, but uNAG activity was severely decreased. Protease inhibitor addition did not preserve uNAG activity. Experimental hematuria, pyuria, and bacteriuria did not seem to affect urinary markers, although further research is needed.

The effect of 24- and 48-hour storage at room temperature on automated total nucleated cell count (TNCC), differential cell count (DCC) and cell morphology was assessed, and the effect of initial total protein concentration on canine and feline body cavity effusion samples (2 to 5 ml) was evaluated. At 24 and 48 hours, TNCC and absolute numbers of neutrophils, macrophages and small lymphocytes were significantly decreased and numbers of unrecognisable cells were significantly increased. Neoplastic cells and intracellular bacteria identified in fresh samples were missed at 24 and 48 hours. The initial total protein concentration was associated with an effect on percentage of unrecognisable cells and small lymphocytes over time. Change in TNCC over time would have resulted in misclassification of the effusion type in four of 47 samples.

Objective: To determine whether use of a closed urine collection system would decrease the incidence of nosocomial bacteriuria in hospitalized dogs, compared with use of an open urine collection system (used, sterile IV bags)., Design: Randomized controlled trial., Animals: 51 hospitalized dogs requiring indwelling urinary catheterization for >or= 24 hours., Procedures: Dogs were randomly assigned to an open or closed urine collection system group. A standardized protocol for catheter placement and maintenance was followed for all dogs. A baseline urine sample was collected via cystocentesis for aerobic bacterial culture, with additional urine samples obtained daily from the urine collection reservoir., Results: 27 dogs were assigned to the open urine collection system group, and 24 were assigned to the closed urine collection system group. The incidence of nosocomial bacteriuria in dogs with open urine collection systems (3/27 [11.1%]) was not significantly different from incidence in dogs with closed urine collection systems (2/24 [8.3%]). Median duration of catheterization was 2 days for dogs in both groups; the range was 1 to 7 days for dogs in the open group and 1 to 5 days for dogs in the closed group., Conclusions and Clinical Relevance: Results suggested that for dogs requiring short-term indwelling urinary catheterization, the type of urine collection system (open vs closed) was not associated with likelihood of developing nosocomial bacteriuria. Use of a strict protocol for urinary catheter placement and maintenance was likely key in the low incidence of nosocomial bacteriuria in the present study.

Objectives: To determine whether serum N-terminal B-type natriuretic peptide (NT-proBNP) concentration in normal dogs, and dogs with heart disease, is affected by freezing, or by sample ageing when stored at room temperature., Animals, Materials and Methods: Thirty six dogs with heart disease and ten normal dogs. Serum NT-proBNP was measured within 60 min of sample collection. Serum was also frozen at -20 degrees C and NT-proBNP measurement was repeated at 1, 24, 48, 72 and 96 h after thawing., Results: Median NT-proBNP increased significantly after freezing (p<0.005) and then progressively decreased at all time points after thawing (p<0.005)., Conclusions: Serum NT-proBNP concentration increases with freezing, and then rapidly decreases over time when stored at room temperature. Concentrations were sufficiently increased after freezing and decreased after 24h at room temperature to affect interpretation. The authors recommend that serum for NT-proBNP assay should be frozen within 1h of sampling and submitted frozen in cold packs., (Copyright 2010 Elsevier B.V. All rights reserved.)

Background: Prior studies failed to detect significant association between hypoalbuminemia and small intestinal lesions., Hypothesis: Use of pictorial templates will enhance consistency of interpathologist interpretation and identification of intestinal lesions associated with hypoalbuminemia., Animals: Tissues from 62 dogs and 25 cats examined as clinical cases at 7 referral veterinary practices in 4 countries., Methods: Retrospective, observational study. Histopathology slides from sequential cases undergoing endoscopic biopsy were examined by 4 pathologists by pictorial templates. Changes for 9 microscopic features were recorded as normal, mild, moderate or severe, and 2- and 4-point scales were tested for consistency of interpretation. Logistic regression models determined odds ratios (OR) of histologic lesions being associated with hypoalbuminemia while kappa statistics determined agreement between pathologists on histologic lesions., Results: There was poor agreement (kappa = -0.013 to 0.3) between pathologists, and institution of origin of slides had effect (kappa = 1.0 for 3 of 4 lesions on slides from Institution 5) on agreement between pathologists on selected histologic features. Using 2 point as opposed to 4-point grading scale increased agreement between pathologists (maximum kappa = 0.69 using 4-point scale versus maximum kappa = 1.0 using 2-point scale). Significant association (P = .019- .04; 95% OR = 3.14-10.84) between lacteal dilation and hypoalbuminemia was found by 3 pathologists., Conclusions and Clinical Importance: Substantial inconsistency between pathologists remains despite use of pictorial template because of differences in slide processing. Distinguishing between mild and moderate lesions might be important source of the disagreement among pathologists.

Background: Cerebrospinal fluid (CSF) is potentially altered by iatrogenic blood contamination at the time of sampling due to the addition of blood-associated leukocytes and protein., Objectives: The objective of this study was to assess whether protein concentration, neutrophil percentage, and the presence of activated macrophages, reactive lymphocytes, or eosinophils in CSF samples with low total nucleated cell concentration (TNCC) are affected by blood contamination or associated with central nervous system (CNS) disease., Methods: Case records from the Royal Veterinary College Diagnostic Laboratory were searched retrospectively for dogs with CSF having < or =5 TNCC/microL. TNCC, RBC, and protein concentrations; neutrophil percentage; and the presence of activated macrophages, reactive lymphocytes, and eosinophils were recorded. Results of magnetic resonance imaging (MRI) also were recorded as a marker of CNS disease., Results: Of 906 cases evaluated, 106 (12%) had blood contamination (>500 RBCs/microL) in CSF. Protein concentration and neutrophil percentage were significantly higher and the presence of eosinophils was more likely in blood contaminated vs noncontaminated samples. Non-blood-contaminated samples with activated macrophages or reactive lymphocytes had higher protein concentrations and neutrophil percentages, and those with activated macrophages were more likely to have a positive finding on MRI., Conclusions: Protein concentration, neutrophil percentage, and the presence of eosinophils are significantly affected by blood contamination in canine CSF having low TNCC. Activated macrophages and reactive lymphocytes are not affected by blood contamination, however, and may be useful in identifying dogs with CNS abnormalities.

A liver biopsy is required to make the diagnosis of most non-vascular liver diseases. Liver biopsy samples can be obtained with several techniques. With proper training and adequate operator experience, the liver biopsy is a safe technique. It is important to evaluate blood coagulation and to use the biopsy in the context of evaluation of the liver and associated structures with ultrasonography, laparoscopy, or other imaging methods. The histological evaluation should be done using international standards as summarized by John Cullen elsewhere in this issue.

A paradigm shift is a fundamental change from a traditional model of thinking. This article presents four paradigm shifts in the diagnoses of urolithiasis, based on the experiences of the authors in the treatment of uroliths in dogs and cats. Case examples are provided to illustrate points regarding urine storage, ultrasonography, use of radiography to assure complete removal of uroliths, and the frequency of upper tract uroliths in cats.

Samples from 75 clinically ill dogs were utilised in the study. APTT and PT tests were performed immediately on fresh citrated plasma samples (Fresh). The remaining plasma was stored at -20 degrees C for less than 4 months (n=36 samples) or between 4 and 7 months (n=39 samples). In batches of five, frozen samples were thawed rapidly and APTT and PT tests were performed on the thawed samples immediately (0RT) and after storage at room temperature (23 degrees C, range: 22-25 degrees C) for 24h (24RT) and 48h (48RT). The median APTT value from the (0RT) samples was significantly longer than that obtained from fresh samples (15s vs. 13.2s) but the PT value was not statistically different (7.8s vs. 7.6s). The median APTT (15s) and PT (7.5s) results from the (24RT) samples were not statistically different to those from the (0RT) samples (APTT: 15s, PT: 7.6s) but both tests were significantly longer (APTT: 16.5s, PT: 9.2s) from the (48RT) samples. We concluded that long term batching and freezing of clinical samples at -20 degrees C is acceptable for measurement of PT but not of APTT. We demonstrated that APTT and PT results do not change following storage of samples at room temperature for 24h but storage for 48h may lead to statistically and clinically significant changes (values at least 25% higher than the high value of the laboratory's reference interval) in both clotting times.

Molecular screening of GM1 gangliosidosis in Shiba dogs was carried out in northern Japan using blood smear specimens after prolonged storage. Of 125 specimens obtained from 3 veterinary teaching hospitals for this screening, 68 specimens (54%) were adequate for direct amplification in a polymerase chain reaction (PCR)-based DNA test, and the percentage of adequacy was different at each hospital (34%, 73%, and 100%), suggesting that the amount of blood on the smear and the storage condition of specimens may affect adequacy. Of the 68 dogs examined, 2 dogs (2.9%) were heterozygous carriers for this disease and the other dogs were all genotypically normal. The results suggest blood smear specimens can be useful for PCR testing after prolonged storage provided specimens contain a generous amount of blood and have been adequately stored. The study also suggests that GM1 gangliosidosis may be widely prevalent in the Shiba dog population in northern Japan.

A 9-year-old spayed female German Shepherd dog with a history of orthopedic disease was presented to the North Carolina State University Veterinary Teaching Hospital for evaluation of recent, progressive, bilateral, hindlimb ataxia. Analysis of cisternal and lumbar cerebrospinal fluid (CSF) samples revealed normal total nucleated cell counts and a mild increase in protein concentration in the lumbar sample. In cytocentrifuged specimens of both CSF samples, aggregates of refractile, angular to irregular, pale blue to colorless, crystalline material were observed in the background. Some of the material appeared birefringent under polarized light. Differentials for the material included contrast agent, epidural anesthetics or other pharmacologic agents, or artifact introduced through sample processing, collection, or handling. Based on investigation of clinical and laboratory processes it was determined that tubes used to collect CSF in the hospital recently had been changed from additive-free glass tubes to silica-coated shatter-resistant plastic tubes (BD Vacutainer Plus serum tubes, silicone-coated, Becton Dickinson). A cytocentrifuged preparation of saline placed in a silica-coated tube contained crystalline material identical to that observed in the CSF samples; saline placed in an additive-free glass tube contained no material. In this case, we document the microscopic appearance of highly concentrated silica particles in cytocentrifuged preparations of CSF and underscore the importance of recognizing and identifying this artifact in cytologic preparations.