Ion-exchange microchromatography and thiobarbituric acid colorimetry for the measurement of canine glycated hemoglobins

Nonenzymatic glycation of hemoglobin is a slow, continuous, and irreversible process which takes place during the whole lifespan of the erythrocyte. When hemolytic diseases are ruled out, the levels of glycated hemoglobins reflect the time-averaged serum glucose concentration for the preceding weeks. Canine hemoglobin also binds physiologically to intraerythrocytic glucose to form a glycated fraction which provides information on the animal's long-term glycemic status. This study describes an overall evaluation of ion-exchange microchromatography and thiobarbituric acid (TBA) colorimetry for the measurement of canine glycated hemoglobins. The intra- and inter-assay coefficients of variation (CVs) found were less than 5% in normal and diabetic canine samples, and both assays proved linear over the analytical range tested, which was wide enough to include the expected clinical values. Under our laboratory's conditions, the reference range for HbA(1) was 5.82 +/- 0.62% and for HbA(1)c was 2.35 -/+ 0.47%. Sample stability was lower using the ion-exchange procedure, with increases in HbA(1) observed after 4 days in whole blood and hemolysates stored at room temperature, after 12 days in whole blood stored at 4 degrees C, and after 7 days in hemolysates stored at 4 degrees C and -20 degrees C. In the case of TBA colorimetry, whole blood was stable for at least 21 days at room temperature and at 4 degrees C, and hemolysates were stable for 18 days at room temperature, at least 21 days at 4 degrees C, and up to 3 months at -20 degrees C.     

Stability of canine factor VIII: coagulant activity in vitro.

A pilot study was undertaken to assess the stability of canine factor VIII:coagulant (FVIII:C) activity over three days, under various storage conditions (plasma at 4, 20 and 37 degrees C, whole blood at 4 and 20 degrees C). Blood collected from normal and hemophiliac dogs was used. Both plasma and whole blood samples appeared to be stable for up to 48 h at 4 and 20 degrees C. A subsequent study evaluated FVIII:C stability at 4 and 20 degrees C when stored as whole blood only. Samples were tested at 0, 24 and 48 h after collection. At 4 degree C there was a significant decline at 24 h (p less than 0.05), from 110% to 97% (mean values). Although the mean value was further decreased at 48 h (89%) this was not significant (p greater than 0.05). No significant change in FVIII:C activity was observed in whole blood stored at 20 degrees C for 24 or 48 h (110% and 107% respectively). These results suggest that canine whole blood samples collected into sodium citrate stored at 20 degrees C are adequate for routine FVIII:C assay for up to 48 h after collection.     

Effect of storage on some constituents of camel serum

SUMMARY Effects of storage at room temperature (23–25°C) and refrigeration (4–5°C) on various biochemical constituents of camel serum were investigated. Albumin, globulin, calcium, phosphorus, cholesterol, aspartate aminotransferase (AST), alkaline phosphatase (AP) and gamma glutamyltransferase (GGT) did not change over 9 days when stored at 4–5°C. At 4–5°C, creatinine, iron and glucose in camel sera remained stable for 6 days; total protein for 7 days; and blood urea nitrogen for 8 days. Decreased activities in creatine kinase (CK), lactate dehydrogenase (LDH) and alanine aminotransferase (ALT) were apparent after 1, 6 and 7 days, respectively. At room temperature, total protein, albumin, globulin, calcium and phosphorus were stable throughout the 9 days. Changes in glucose and iron occurred after 3 days. Stability at room temperature for LDH was 1 day; AST, 3 days; GGT and ALT, 6 days; and AP, 8 days. CK activity had already declined by 4 hours and by 9 days, only 34% activity remained.     

Effect of serum storage, anti-inflammatory oral doses of prednisone, and spontaneous hyperadrenocorticism on serum glutamate dehydrogenase activity in dogs

BACKGROUND: Glutamate dehydrogenase (GLDH) is a mitochondrial enzyme with highest activity in periacinar hepatocytes. It is reported to be a sensitive indicator of hepatic injury; however, results of studies regarding tissue specificity are contradictory. OBJECTIVES: The purpose of the study reported here was to examine the effect of 3 factors on serum GLDH activity in dogs: serum storage, anti-inflammatory oral doses of prednisone, and spontaneous hyperadrenocorticism (HAC). METHODS: Stability of enzyme activity was determined by comparing serum samples stored at approximately 20 degrees C, 4 degrees C, and 20 degrees C for 4, 24, 48, and 72 hours, 1 week, and 6 months. To determine whether orally administered prednisone affected GLDH activity, the median difference in serum GLDH activity was compared between 5 untreated control dogs and 8 dogs that had received a tapering oral dose of prednisone. Lastly, GLDH enzyme activity was compared between 17 dogs with HAC and 16 age-matched controls. RESULTS: GLDH activity remained stable for 48 hours, 1 week, and 6 months, in serum stored at approximately 20 degrees C, 4 degrees C, and 20 degrees C, respectively. The median change in GLDH activity was not significantly different between dogs receiving prednisone and controls; however, dogs with HAC had significantly higher values than those of age-matched controls. CONCLUSIONS: Serum samples should be maintained at 4 degrees C if analysis of GLDH activity will be delayed by >48 hours; serum stored at 20 degrees C yields reliable results for up to 6 months. Serum GLDH activity was not increased in most dogs receiving short-term, anti-inflammatory oral doses of prednisone, in contrast to its increased activity in dogs with HAC.     

The effect of storage conditions on samples for the evaluation of copper status in blesbok (Damaliscus pygargus phillipsi)

Investigaltions to determine the effect of sample storage on the concentration of copper in liver tissue and on the activity of erythrocyte superoxide dismutase were undertaken in preparation for a study of blesbok (Damaliscus pygargus phillipsi) that were suspected to be suffering from copper deficiency. Two liver samples were collected from each of 20 culled blesbok in a manner that simulated the collection of biopsies from the live animal. These samples were stored either in 10% formalin or frozen at -20 degrees C until analysed 4 1/2 months later. The effect of different methods of sample storage on superoxide dismutase activity was determined. Erythrocytes collected from 3 Jersey cows and 5 culled blesbok were washed and divided into 0.5 ml portions, stored at room temperature (approximately 20 degrees C), in a refrigerator (4 degrees C), frozen at -20 degrees C in a freezer, and in liquid nitrogen (-200 degrees C). An analysis of superoxide dismutase activity was undertaken using a commercial assay kit at intervals of 2-4 days until the levels of activity had fallen significantly. The copper concentration in formalin-preserved liver samples was significantly lower than that measured in frozen liver tissue apparently as a result of leaching. The activity of superoxide dismutase in cattle blood was unchanged for 4 days at room temperature but fell appreciably after 2 days at 4 degrees C and -20 degrees C. Enzyme activity remained unchanged for 200 days in erythrocytes stored in liquid nitrogen. Superoxide dismutase activity levels in healthy blesbok were considerably lower than those measured in Jersey cows and remained unaffected for up to 6 days in samples stored at 4 degrees C and 20 degrees C. The level of activity fell significantly thereafter. Samples stored in liquid nitrogen were unchanged after 40 days.     

Measurement of urinary glycosaminoglycans in dogs

OBJECTIVES: To measure urine concentrations of sulfated glycosaminoglycans (GAGs), determine optimal storage conditions for urine samples, establish a reference range, and determine whether there is correlation between 24-hour total urine GAG excretion and the GAG-to-creatinine ratio (GCR). ANIMALS: 14 healthy adult dogs. PROCEDURE: Single urine sample GAG concentrations and GCRs were measured in samples collected from 14 healthy dogs at the start of the 24-hour collection period. Twenty-four-hour total urine GAG excretions were determined from urine collected during a 24-hour period in the same 14 dogs. Total sulfated GAG concentrations were also measured in urine from these dogs after the urine had been stored at 4 degrees C and -20 degrees C for 1, 7, and 30 days. RESULTS: Urine GAG concentrations were not significantly different from baseline values after urine was stored at 4 degrees C for up to 1 day and -20 degrees C for up to 30 days. Neither single urine sample GAG concentration (R2, 0.422) nor GCR (R2, 0.084) was an adequate predictor of 24-hour total urine GAG excretion. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study provide data that can be used to establish a reference range for 24-hour total urine GAG excretion in dogs and adequate conditions for sample storage. Contrary to findings in humans, there was no significant linear correlation between 24-hour total urine GAG excretion and single urine sample GCR in dogs, limiting clinical use of the single urine sample test.     

A rapid microassay for measuring the luminol-dependent chemiluminescent response in canine whole blood

A microassay for the luminol-dependent chemiluminescence (CL) response in canine whole blood was developed to measure indirectly the oxidative metabolism of peripheral blood leukocytes. Fifty microliters of blood were mixed with 705 microliters of Hank's balanced salt solution containing 25 mM Hepes and 1.3 x 10(-4) M luminol. This mixture was allowed to equilibrate for 5 min after which 60 microliters of latex beads (0.801 microns diameter) were added as a stimulant, and the CL response was monitored continuously for 5 min at 37 degrees C using a luminometer. The whole blood CL response was significantly correlated (r = 0.784, P less than 0.01, n = 14) with the number of neutrophils in the peripheral blood. Further, the whole blood CL response was abolished by the depletion of neutrophils after passing the blood through an adherence column and by the addition of sodium azide. The relative chemiluminescent light unit (RCLU) was a reliable marker for comparing each peak value in different samples. The coefficient of variation (CV) of repetitive samples was 9.87%, and the CV of 14 normal dogs was 15.7%. This method is useful and applicable for screening the CL response in canine whole blood.     

Changes in Plasma Progesterone Levels During Storage of Heparinized Whole Blood from Cow,Horse, Dog and Pig

Progesterone concentrations in heparinized plasma harvested immediately after blood collection were compared with levels obtained after storage of the corresponding whole blood for 2 h, 4 h, 6 h, 1 day, 2 days and 5 days at room temperature and in a refrigerator. The blood was taken during the luteal phase from 4 dogs, 4 horses, 4 pigs and 8 cows. For 4 cows the storage time was extended to 9 and 20 days. No significant effect of whole blood storage time on plasma progesterone concentrations could be shown for dogs or pigs. For the horse a slight but significant decrease was demonstrated when the blood was kept at room temperature. For the cow, however, a dramatic decrease was observed even when the blood was stored in the refrigerator. Following incubation of cow’s blood at room temperature, progesterone levels were close to zero after 1–2 days. By further extending the storage period, a reappearance of assayable progesterone could be elicited. For all species it was found that the storage of plasma at room temperature for 5–9 days did not change the progesterone concentrations.     

Artifactual changes in PCV, hemoglobin concentration, and cell counts in bovine, caprine, and porcine blood stored at room and refrigerator temperatures

BACKGROUND: Blood samples collected from farm animals for hematology testing may not reach the laboratory or be examined immediately upon collection, and in some cases may need to be transported for hours before reaching a laboratory. OBJECTIVE: The objective of this study was to investigate the artifactual changes that may occur in PCV, hemoglobin (Hgb) concentration, and cell counts in bovine, caprine, and porcine blood samples stored at room (30 degrees C) or refrigerator (5 degrees C) temperature. METHODS: Baseline values for PCV, Hgb concentration, and RBC and WBC counts were determined immediately after blood collection from 36 cattle, 32 goats, and 48 pigs using manual techniques. Blood samples were split into 2 aliquots and stored at 30 degrees C or 5 degrees C. Hematologic analyses were carried out at specified intervals during 120 hours of storage. Results were analyzed by repeated measure ANOVA; results at different temperatures were compared by paired t-tests. RESULTS: Compared to baseline values, there were no significant changes in Hgb concentration, RBC count, or WBC count in samples from cattle; in Hgb concentration and RBC count in samples from goats; and in Hgb concentration and WBC count in samples from pigs throughout the 120 hours of storage at both 30 degrees C and 5 degrees C. Significant changes (P <.05) from baseline occurred in PCV after 14 hours of storage at 30 degrees C and after 19 hours of storage at 5 degrees C in cattle and goats; and after 10 hours of storage at 30 degrees C and 14 hours of storage at 5 degrees C in pigs. Significant changes also were observed in Hgb concentration at 96 hours at 30 degrees C and 5 degrees C, and in RBC counts at 48 hours at 30 degrees C and 96 hours at 5 degrees C in porcine samples; and in total WBC counts at 120 hours at 30 degrees C and 5 degrees C in caprine samples. Artifactual changes were more pronounced in the samples stored at 30 degrees C. CONCLUSIONS: At both 30 degrees C and 5 degrees C, blood samples from cattle and goats can be stored for up to 12 hours, while blood samples from pigs can be stored for up to 8 hours without any significant changes in PCV. Blood samples from all 3 species can be stored for more than 24 hours without significant changes in Hgb concentration, RBC count, and total WBC count.     

Validation of a species-optimized enzyme-linked immunosorbent assay for determination of serum concentrations of insulin in dogs

Background: Measurement of canine serum insulin has relied on methods developed to measure human insulin. A species‐optimized test for measurement of serum insulin in dogs is now commercially available. Objective: The purpose of this study was to validate the canine ELISA for determination of serum insulin concentration in dogs. Methods: Precision was determined by evaluating intra‐ and interassay coefficient of variation (CV), and accuracy was determined by dilution and spike recovery studies. A method comparison study with samples from 34 clinically healthy dogs and 73 dogs examined for various illnesses and disorders (“patients”) was performed using the canine ELISA and an ELISA for human insulin. Biologic relevance of the canine assay was evaluated by measuring insulin in samples collected from 8 healthy dogs after administration of glucagon. A stability study was preformed with 6 samples stored at 20°C, 4–8°C, and ?20°C. Results: For the canine ELISA, intra‐ and interassay CVs were 4.3–7.8% and 4.4–7.7%, respectively. Mean recovery after dilution was 99% and recovery after spiking with porcine insulin was 116%. The canine and human ELISAs correlated well (r²=.94 for healthy dogs, r²=.88 for patient samples). After glucagon injection serum insulin concentrations increased significantly in 8 dogs. Insulin was stable for 30 days in 6 serum samples stored at ?20°C and in most samples for 8 days at 4–8°C. Insulin was stable for <3 days at room temperature (20°C). Conclusions: The new canine serum insulin ELISA had good precision and accuracy and correlated well with the previously used assay.     

Human-parathormone assay for use in dogs: validation, sample handling studies, and parathyroid function testing

Ten commercially available parathormone (PTH) assays were competitively validated, using dilutional parallelism, intra-assay and interassay coefficients of variation, and sensitivity and measured responses of 2 dogs to calcium and EDTA infusions. A 2-site immunoradiometric assay for intact human-PTH was superior to the others for estimating canine-PTH, met the criteria for validity, and was further investigated. A series of sample-handling studies was performed. Serum and plasma samples stored at 24 C lost 15% (n = 5; P less than 0.05) of PTH between 2 and 24 hours. This did not occur at 6 C. The mean PTH concentration of sera from blood samples clotted at 24 C was 6% (P less than 0.05) higher than equivalent EDTA samples. Serum samples stored at 6 and 37 C deteriorated 35% and 100% (n = 5; P less than 0.05), respectively, after 1 week, whereas samples stored at -20 and -70 C for 4 weeks did not deteriorate. There was no significant deterioration of PTH in samples frozen (-40 C) and thawed up to 7 times (n = 5). Parathyroid function testing was investigated by use of 2-hour infusions of disodium EDTA (25 mg/kg/h), 10-minute infusions of calcium gluconate (3 mg of elemental calcium/kg/10 min), and physiologic saline controls (n = 8). Renal function was monitored before and after EDTA infusion by exogenous creatinine clearance. Infusion of disodium EDTA increased mean PTH concentration from 67 (time 0) to 317 and 235 pg/ml at 90 and 180 minutes, respectively (P less than 0.001). Infusion of calcium gluconate decreased mean PTH concentration from 84 (time 0) to 14 and 12 pg/ml at 15 and 60 minutes, respectively (P less than 0.005). There were no observable side effects of the infusions in normal conscious dogs and no differences in exogenous creatinine clearance after EDTA infusion.     

Factors affecting the survival of Streptococcus suis type 2

The survival of Streptococcus suis type 2 was assessed in experimentally inoculated faeces and dust stored at 0, 9 and 22 to 25 degrees C. The organism survived in faeces for 104 days at 0 degrees C, up to 10 days at 9 degrees C and up to eight days at 22 to 25 degrees C. It survived in dust for up to 54 days at 0 degrees C and up to 25 days at 9 degrees C but could not be isolated from dust stored at room temperature for 24 hours. The organism survived at 4 degrees C in nutrient medium for up to nine months but in distilled water for only one to two weeks. At 50 degrees C it survived in water or broth for up to two hours but at 60 degrees C it only survived for 10 minutes. The organism was rapidly inactivated by disinfectants and cleansers, commonly used on farms and in laboratories, at concentrations less than those recommended for use by the manufacturers.     

Detection of Taenia hydatigena copro-antigens by ELISA in dogs

A sandwich-ELISA was developed for the detection of soluble Taenia hydatigena antigens in fecal samples of dogs. Affinity-purified polyclonal catching antibodies and alkaline phosphatase-conjugated detecting antibodies were employed, which had been obtained from rabbits hyperimmunized with excretory/secretory antigens derived from in vitro maintained adult Taenia hydatigena. The assay allowed the detection of 800 ng T. hydatigena antigen g-1 of feces as a lower limit. Six helminth-free dogs were each infected with 10 T. hydatigena cysticerci isolated from Swiss sheep. After prepatent periods ranging from 57 to 71 days, the dogs started to excrete Taenia eggs and/or proglottids. The ELISA detected Taenia antigens in all six dogs during the prepatent period starting individually between Day 18 and 45 post-infection (p.i.). Anthelmintic treatment of three dogs at Day 95 p.i. resulted in elimination of the cestodes and within the 5 following days in the disappearance of Taenia antigens from feces. The specificity of the assay was evaluated by testing crude antigens derived from helminths or bacteria. Four Taenia species showed cross-reactivity at concentrations of 5 micrograms protein ml-1. Conversely, no cross-reactions occurred with various antigen batches derived from Echinococcus granulosus, E. multilocularis, Dipylidium caninum, Mesocestoides corti, Diphyllobothrium sp., Toxocara canis and bacterial antigens (Salmonella and Escherichia). Moreover, fecal samples from dogs naturally infected with T. canis (n: 13), hookworms (n: 2), Trichuris vulpis (n: 13) and of 10 dogs with mixed infections with these three nematode groups were tested, and results confirmed the high degree of specificity. The Taenia antigens detectable by this ELISA remained immunologically stable in native feces stored at +25 degrees, +4 degrees or at -20 degrees C for at least 5 days.     

Artifactual changes in canine blood following storage, detected using the ADVIA 120 hematology analyzer

BACKGROUND: Artifactual changes in blood may occur as a consequence of delayed analysis and may complicate interpretation of CBC data. OBJECTIVE: The aim of this study was to characterize artifactual changes in canine blood, due to storage, using the ADVIA 120 hematology analyzer. METHODS: Blood samples were collected into EDTA from 5 clinically healthy dogs. Within 1 hour after blood sample collection and at 12, 24, 36 and 48 hours after storage of the samples at either 4 degrees C or room temperature (approximately 24 degrees C), a CBC was done using the ADVIA 120 and multispecies software. A linear mixed model was used to statistically evaluate significant differences in values over time, compared with initial values. RESULTS: The HCT and MCV were increased significantly after 12 hours of collection at both 4 degrees C and 24 degrees C, and continued to increase through 48 hours. The MCHC initially decreased significantly at 12-24 hours and then continued to decrease through 48 hours at both temperatures. Changes in HCT, MCV, and MCHC were greater at 24 degrees C than at 4 degrees C at all time points. A significant increase in MPV and a decrease in mean platelet component concentration were observed at all time points at 24 degrees C. Samples stored at 24 degrees C for 48 hours had significantly higher percentages of normocytic-hypochromic RBCs, and macrocytic-normochromic RBCs, and lower platelet and total WBC counts. CONCLUSIONS: Delayed analysis of canine blood samples produces artifactual changes in CBC results, mainly in RBC morphology and platelet parameters, that are readily detected using the ADVIA 120. Refrigeration of specimens, even after 24 hours of storage at room temperature, is recommended to improve the accuracy of CBC results for canine blood samples.     

Comparison of methods for assay of fecal proteolytic activity

Fecal proteolytic activity (FPA) in ten normal dogs was readily detected either calorimetrically using azocasein substrate or by radial enzyme diffusion into agar gels containing casein substrate. Daily FPA ranged from 17 to 207 azocasein units/g (ACUIG) or 4 to 18 mm of casein hydrolysis, while mean 3-day FPA ranged from 58 to 10 1 ACUIG or 7 to 15 mm. Studies of proteolytic activity remaining after treatment of fecal extracts with a specific trypsin inhibitor indicated that trypsin accounted for 0% to 71% of proteolytic activity. Proteolytic activity decreased steadily in fecal specimens stored at room temperature or above, but there was only slight loss in activity during storage for up to 5 days at 4 degrees C. Proteolytic activity was unaffected by repeated freezing and thawing and samples could be stored for long periods at -2 degrees C without noticeable loss of activity. It is concluded that assays of FPA using either azoprotein substrate or radial enzyme diffusion into agar gels containing casein substrate allow evaluation of exocrine pancreatic function in dogs, provided that several samples are tested. These methods are suitable for application in a variety of species.     

Effects of hemolysis and storage on quantification of hormones in blood samples from dogs, cattle, and horses

Veterinary diagnostic endocrinology laboratories frequently receive hemolyzed plasma, serum, or blood samples for hormone analyses. However, except for the previously reported harm done by hemolysis to canine insulin, effects of hemolysis on quantification of other clinically important hormones are unknown. Therefore, these studies were designed to evaluate effects of hemolysis on radioimmunoassay of thyroxine, 3,5,3'-triiodothyronine, progesterone, testosterone, estradiol, cortisol, and insulin in equine, bovine, and canine plasma. In the first experiment, hormones were measured in plasma obtained from hemolyzed blood that had been stored for 18 hours. Blood samples were drawn from pregnant cows, male and diestrous female dogs, and male and pregnant female horses. Each sample was divided into 2 equal portions. One portion was ejected 4 times with a syringe through a 20-gauge (dogs, horses) or 22-gauge (cows) hypodermic needle to induce variable degrees of hemolysis. Two subsamples of the blood were taken before the first and after the first, second, and fourth ejections. One subsample of each pair was stored at 2 to 4 C and the other was stored at 20 to 22 C for 18 to 22 hours before plasma was recovered and stored at -20 C. The second portion of blood from each animal was centrifuged after collection; plasma was recovered and treated similarly as was blood. Concentrations of thyroxine in equine plasma, of 3,5,3'-triiodothyronine, estradiol, and testosterone in equine and canine plasma, and of cortisol in equine plasma were not affected by hemolysis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Viability and infectivity of Trichinella spiralis muscle larvae in frozen horse tissue

Many aspects of the biology and epidemiology of Trichinella infection in the horse are poorly understood, including survival of Trichinella spp in horse muscle. In this study, we have assessed the freeze tolerance of T. spiralis in horse meat stored at 5, -5, and -18 degrees C for 1 day to 24 weeks. Results demonstrate a steady reduction in the number of live ML recovered from the cold stored meat samples. On Day 1, recovery of live larvae had been reduced by 18.6%, 50.1%, and 37.2%, and by 4 weeks, recovery of larvae had been reduced by 65.4%, 66.5%, and 96.2% in samples stored at 5, -5, and -18 degrees C, respectively. Infectivity results (measured as reproductive capacity index (RCI)) from mice inoculated with larvae recovered from non-frozen meat samples at day 0 was 23.5. Following storage at -18 degrees C for one and two days, the RCIs were 2.09 and 0.99, respectively. Small numbers of infective larvae were still present in meat samples stored at -18 degrees C for 4 weeks. The RCI of ML recovered from meat samples stored at -5 degrees C was 14.99 and 6.36 at 2 weeks and 4 weeks respectively; the RCI of samples stored at 5 degrees C was 23.1 at 8 weeks, and fell rapidly thereafter (12 week RCI 1.33; 0 at 24 weeks). These data demonstrate that infective T. spiralis, a non-freeze tolerant species, can survive for at least 4 weeks in horse tissue frozen at -5 or -18 degrees C, and that the numbers of infective larvae decrease substantially by day 2 at -18 degrees C and by week 4 at -5 degrees C.     

Prevalence and characterization of urinary tract infections in dogs with surgically treated type 1 thoracolumbar intervertebral disc extrusion

OBJECTIVE: To determine the prevalence of urinary tract infections (UTI), factors that correlate positively with UTI, and whether identified UTI are most likely community- or hospital acquired in dogs with surgically treated type 1 thoracolumbar intervertebral disc (IVD) extrusions. STUDY DESIGN: Prospective cross-sectional clinical study. SAMPLE POPULATION: Dogs (n=92) that were surgically treated for a thoracolumbar extradural compressive spinal cord lesion that was consistent with type 1 IVD extrusion. METHODS: Dogs were evaluated for bacterial lower UTI when possible by cystocentesis and urine culture before surgery, and 48-72, 96-120 hours, and 7 days after surgery while hospitalized. Paraparesis, confirmation of thoracolumbar extruded nucleus pulposus, and informed owner consent were required for study inclusion. Urine specimens (n=297) were cultured and both objective and subjective clinical data were obtained. RESULTS: Prevalence of UTI in dogs with surgically treated type 1 thoracolumbar IVD extrusion was 27% (25 dogs). Temporal prevalence of UTI was 15% (13/89) before surgery, 12% (11/91) at 2-3 days, 16% (12/76) at 4-5 days, and 20% (8/41) at 7 days after surgery. Statistically significant factors affecting UTI prevalence included neurologic and urinary status, sex, administration of perioperative antibiotics, and amount of time body temperature was <35 degrees C during anesthesia. CONCLUSION: UTI are common in dogs with surgically treated type 1 thoracolumbar IVD extrusion. Females, dogs that cannot ambulate or voluntarily urinate, dogs not administered perioperative cefazolin, and dogs whose body temperature falls <35 degrees C during anesthesia have a higher incidence of UTI. CLINICAL RELEVANCE: All dogs with surgically treated type 1 thoracolumbar IVD extrusion should be monitored for the presence of UTI; however, close attention should be paid to females and dogs that cannot ambulate or voluntarily urinate.     

Stability of vitamin E in blood and plasma from cattle, sheep, and pigs.

The stability of alpha-tocopherol concentrations in sheep, cattle, and pig blood and plasma stored at different temperatures was examined. For all species, the vitamin was stable for at least 6 days in plasma stored at -20 C, 4 C, and 25 C and in blood stored at 4 C and 25 C. For sheep and cattle, the vitamin was stable for at least 6 days in plasma stored at 37 C, but it was unstable in blood from all species stored at 37 C and in pig plasma stored at 37 C.     

Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation.

Although Clostridium difficile is recognized as a cause of enterocolitis in horses and humans, there has been little work published regarding the lability of C. difficile and its toxins in feces. A significant decrease in recovery of C. difficile from inoculated equine fecal samples occurred during storage. Recovery after storage in air at 4 degrees C decreased from 76% (37/49) after 24 hours to 67% (33/49) at 48 hours and 29% (14/ 49) after 72 hours. In contrast to aerobic storage, 25 of 26 samples stored anaerobically at 4 degrees C yielded growth of C. difficile for 30 days, whereas the organism was only detected for 2.5 +/- 2.52 days (x +/- SD) in paired samples stored aerobically. The use of an anaerobic transport medium was effective in maintaining viability of C. difficile. These findings indicate that poor aerotolerance is the reason for the rapid decrease in culture yield. In contrast to C. difficile organisms stored aerobically at 4 degrees C, C. difficile toxins were considerably more stable and could be detected by enzyme-linked immunosorbent assay in both broth and inoculated fecal samples for at least 30 days. The poor survival of C. difficile but the stability of its toxins when feces are stored aerobically must be considered when submitting samples for diagnosis of C. difficile-associated enterocolitis in horses and when interpreting laboratory results.