Automated Blood Cell Counting Systems: A Comparison of the Coulter S-Plus IV, Ortho ELT-8/DS, Ortho ELT-8/WS, Technicon H-1, and Sysmex E-5,000

Four automated blood cell counting systems were evaluated at the Michigan State University Veterinary Clinical Center (VCC) for their suitability for analyzing various animals' blood in a university teaching hospital laboratory. The instruments were compared to a Coulter Model S-Senior (Coulter Electronics)which had been used for 8 years, and was to be replaced. The instruments were a Coulter Model S-Plus IV, an Ortho ELT-8/ds (Ortho Diagnostics Systems), a Technicon H-1 (Technicon Instruments), and a Sysmex E-5,000 (Toa Medical Electronics). Additionally, an Ortho ELT-8/ws at a private laboratory (Cenvet) was compared to the Coulter S-Senior at the VCC. Based on these evaluations, only the Sysmex E-5,000 was considered unacceptable for the VCC laboratory. Certain advantages and disadvantages of the instruments are described in this article. The comparisons among instruments were not as consistent or repetitive as expected for a controlled experiment but did provide information likely useful to others considering using or purchasing an automated blood cell counting system for a veterinary laboratory.     

Qualitative and quantitative comparison of renal vs. hepatic ultrasonographic intensity in healthy dogs

A prospective, cross-sectional study was performed to qualitatively and quantitatively compare the echogenicity of the renal cortex relative to the liver in healthy dogs. Twenty-five normal adult dogs were examined ultrasonographically. Three standard B-mode images (8.0 MHz) and three tissue harmonic images of the cranial pole of the right kidney adjacent to the caudate lobe of the liver were obtained. Renal and hepatic echogenicities were qualitatively compared by two observers. Subsequently, regions of interest (ROIs) were drawn in the renal cortex and the adjacent liver parenchyma at equal depths on each image, using two different ROI geometries: deep adjacent half-annular ROIs centered at the focal zone and small superficial adjacent squares placed in the near field. Renal and hepatic mean pixel intensities were quantified and averaged for individual subjects. Qualitatively, the right renal cortex was more commonly hyperechoic to liver. Quantitatively, the renal cortical mean pixel intensity was significantly higher than that of liver using deep half-annular ROIs, but not superficial square ROIs, for both standard (P = 0.0007) and harmonic (P = 0.0107) tissue imaging. These findings suggest that the renal cortex can be slightly hyperechoic to adjacent liver. The framework within which the canine renal cortical parenchyma is routinely evaluated in abdominal ultrasonography should be reconsidered, and mild hyperechogenicity relative to the liver (at 8.0 MHz) interpreted as a normal finding.     

Serum bile acids: reference values in healthy dogs and comparison of two kit methods

Serum bile acid (SBA) reference intervals were established by use of a radioimmunoassay method for fasting dogs to be 0.2 to 4.3 micro mol/L (n = 60) and for 2 hour postprandial samples to be 0.6 to 24.2 micro mol/L (n = 37). The SBA reference intervals estimated using an enzymatic method were 0 to 8.6 micro mol/L for fasting (n = 26) and 0 to 29.8 micro mol/L for 2 hour postprandial samples (n = 36). The correlation between the two methods including samples from healthy dogs and clinical cases is good (n = 128, r = 0.82, p < 0.0001). The radioimmunoassay method is linear to 50 micro mol/L and the enzymatic method is linear to 100 micro mol/L, thus both methods require serum dilutions to be made in many cases of primary liver disease. The enzymatic method is less expensive and more convenient for use in a clinical laboratory but requires a greater sample volume (400 micro I) than the RIA method (50 micro I). Both methods have adequate precision and accuracy to be useful as diagnostic tests of liver function in dogs.     

Creatinine concentration in plasma from dog, rat, and mouse: a comparison of 3 different methods

BACKGROUND: There are numerous methods for analyzing creatinine concentration in plasma, including the Jaffé alkaline picrate method in various modifications, enzymatic tests, and chromatographic methods. OBJECTIVE: The purpose of this study was to evaluate whether an enzymatic method could replace a Jaffé method for routine creatinine measurements in plasma from dogs, rats, and mice. The enzymatic method and a compensated Jaffé method were tested against a high-pressure liquid chromatography (HPLC) method, regarded as the gold standard for creatinine measurement. METHODS: Heparinized plasma samples were obtained from 20 beagle dogs, 20 Wistar rats, and 20 CD1-strain mice. The 2 test kits (Roche Diagnostics), Creatinine Jaffé Compensated and the enzymatic Creatinine Plus Version 2 reagent, were used on a Cobas Integra 400. The Jaffé compensated method used a calibration adjustment of 18 micromol/L to correct for the protein matrix in serum and plasma. The HPLC method was an isocratic method using a weak cation-exchange column following protein precipitation. RESULTS: Creatinine concentrations obtained using the enzymatic and the Jaffé methods differed significantly from the results obtained by the HPLC method. For dog plasma, mean values of 61.2, 61.8, and 67.8 micromol/L were obtained by the compensated Jaffé, enzymatic and HPLC methods, respectively. In the rat, respective mean values were 26.7, 21.9, and 23.0 micromol/L, and in the mouse, respective mean values were 14.2, 5.4, and 9.2 micromol/L. CONCLUSION: The enzymatic method can replace the Jaffé method for plasma creatinine determination in dogs, rats, and mice because results from the enzymatic method were closer to HPLC values than were those of the Jaffé method.     

Histological dermal changes caused by preparation and application procedures in percutaneous dose toxicity studies in dogs,rabbits and rats

We reevaluated histological slides of dorsal skin in control animals from past percutaneous dose toxicity studies using dogs, rabbits and rats to provide background data concerning histological changes related to preparation and application procedures and vehicles or embrocations of every variety. Acanthosis, dermal or perifollicular inflammatory cell infiltration in dogs; hyperkeratosis, acanthosis, dermal inflammatory cell infiltration or hemorrhage in rabbits; and acanthosis, dermal inflammatory cell infiltration, crust or foreign body granuloma in rats were present as procedure-related underlying histological changes in the control animals. Four mechanical acts, (1) rubbing with gauze to remove an administered substance for reapplication, (2) use of a taut bandage to avoid slipping from the application site, (3) peeling a patch off as a preparation procedure for reapplication, and (4) clipping or shaving, were considered to cause injury to the skin. The degree of influence of the various application procedures was found to be as follows: sham, lotion < cream < ointment and tape in dogs; untreated control, sham < lotion < tape and poultice in rabbits; and sham, sodium carboxymethylcellulose < olive oil and lotion < ointment and tape in rats. The degree of ointment influence on rabbits is equivocal.     

Relation between age, body weight, and medial retropharyngeal lymph node size in apparently healthy dogs.

The aims of this study were to determine the size of the medial retropharyngeal lymph nodes in apparently healthy dogs using ultrasonography and to investigate relationships between body weight (1.8-59 kg), age (1.0-15 years), and medial retropharyngeal lymph node sizes (width, height, and length). The sample population consisted of 100 apparently healthy, volunteered, adult dogs. The data were normally distributed, thus mean, SD, and Pearson's correlation were used. Repeatability of ultrasound measures was assessed as the percentage of differences between duplicate measures that were within 2 SDs of the differences: all measures were at least 93% repeatable (differences typically were < or = 0.25 cm and always < 1 cm). No difference between sexes was observed. The medial retropharyngeal lymph node increased in size with increased body weight (r = 0.46 to 0.59) and decreased in size with increased age (r = -0.30 to -0.50). Although statistically significant, the actual variation is not likely clinically important due to the small range of sizes, measurement error, and various combinations of age and body weight. Therefore, regardless of body weight or age, the average width is 1.0 cm, height is 0.5 cm, and length is 2.5 cm and maximum width is 2 cm, height is 1 cm, and length is 5 cm. Based on the maximal difference between duplicate measures (with some exception), any change > or = 0.4 cm with width or height, or > or = 1.0 cm in length, in a follow up measurement probably represents a true biological change rather than measurement error.     

Contrast‐enhanced voiding urosonography: A new,radiation‐free,alternative method for imaging of urinary bladder and urethra in healthy dogs

Contrast‐enhanced voiding urosonography (CE‐VUS) has been generally considered as a promising tool to diagnose vesicoureteral reflux and abnormalities in lower urinary tract in human patients, especially in children. The purpose of this prospective study is to evaluate the quality of images of the urinary bladder and urethra obtained by CE‐VUS using a second‐generation ultrasound contrast agent (SonoVue®) in healthy dogs and to investigate the safety profile of SonoVue® after intravesical administration. Eighty‐four CE‐VUS examinations with SonoVue® were successfully performed in both unsedated (39/84) and sedated (45/84) dogs. Contrast‐enhanced voiding urosonography examination of urinary bladder was technically successful in all (84/84) dogs. The image quality was not considered adequate in five (5/84) dogs including three dogs in whom layering of contrast media during filling phase was observed and two dogs with premature destruction of microbubbles. In these five dogs, the problem was readily recognized and corrected such that the procedure was still successfully undertaken. The assessment of the urethra during spontaneous micturition was successfully performed in all (84/84) dogs in whom voiding was elicited during the examination. No side effects were observed after intravesical application of SonoVue®. This study demonstrates that CE‐VUS is a feasible and valuable technique to evaluate low urinary tract morphology and function in dogs. Based on our review of the literature, there are no published reports about the use of this method in dogs.     

Validation of the Sysmex XT‐2000iV hematology system for dogs,cats, and horses. I. Erythrocytes,platelets, and total leukocyte counts

Background: The Sysmex XT‐2000iV is a laser‐based, flow cytometric hematology system that has been introduced for use in large and referral veterinary laboratories. Objective: The purpose of this study was to validate the Sysmex XT‐2000iV for counting erythrocytes, reticulocytes, platelets, and total leukocytes in blood from ill dogs, cats, and horses. Methods: Blood samples from diseased animals (133 dogs, 65 cats, and 73 horses) were analyzed with the Sysmex XT‐2000iV and the CELL‐DYN 3500. Manual reticulocyte counts were done on an additional 98 canine and 14 feline samples and manual platelet counts were done on an additional 73 feline and 55 canine samples, and compared with automated Sysmex results. Results: Hemoglobin concentration, RBC counts, and total WBC counts on the Sysmex were highly correlated with those from the CELL‐DYN (r≥0.98). Systematic differences occurred for MCV and HCT. MCHC was poorly correlated in all species (r=0.33–0.67). The Sysmex impedance platelet count in dogs was highly correlated with both the impedance count from the CELL‐DYN (r=0.99) and the optical platelet count from the Sysmex (r=0.98). The Sysmex optical platelet count included large platelets, such that in samples from cats, the results agreed better with manual platelet counts than with impedance platelet counts on the Sysmex. Canine reticulocyte counts on the Sysmex correlated well (r=0.90) with manual reticulocyte counts. Feline reticulocyte counts on the Sysmex correlated well with aggregate (r=0.86) but not punctate (r=0.50) reticulocyte counts. Conclusion: The Sysmex XT‐2000iV performed as well as the CELL‐DYN on blood samples from dogs, cats, and horses with a variety of hematologic abnormalities. In addition, the Sysmex detected large platelets and provided accurate reticulocyte counts.     

Validation of the Sysmex XT‐2000iV hematology system for dogs,cats, and horses. II. Differential leukocyte counts

Background: The Sysmex XT‐2000iV is a laser‐based, flow cytometric hematology system that stains nucleic acids in leukocytes with a fluorescent dye. A 4‐part differential is obtained using side fluorescence light and laser side scatter. Objective: The purpose of this study was to validate the Sysmex XT‐2000iV for determining differential leukocyte counts in blood from ill dogs, cats, and horses. Methods: Blood samples from diseased animals (133 dogs, 65 cats, and 73 horses) were analyzed with the Sysmex XT‐2000iV (Auto‐diff) and the CELL‐DYN 3500. Manual differentials were obtained by counting 100 leukocytes in Wright‐stained blood smears. Results: Leukocyte populations in the Sysmex DIFF scattergram were usually well separated in equine samples, but were not as well separated in canine and feline samples. Correlation among the Sysmex XT‐2000iV, CELL‐DYN 3500, and manual counts was excellent for neutrophil counts (r ≥.97) and good for lymphocyte counts (r ≥.87) for all three species. Systematic differences between the 3 methods were seen for lymphocyte and monocyte counts. The Sysmex reported incomplete differential counts on 18% of feline, 13% of canine, and 3% of equine samples, often when a marked left shift (>10% bands) and/or toxic neutrophils were present. Eosinophils were readily identified in cytograms from all 3 species. Neither the Sysmex nor the CELL‐DYN detected basophils in the 7 dogs and 5 cats with basophilia. Conclusions: The Sysmex XT‐2000iV automated differential leukocyte count performed well with most samples from diseased dogs, cats, and horses. Basophils were not detected. Immature neutrophils or prominent toxic changes often induced errors in samples from cats and dogs.     

Surgery: Comparison of three closure methods and two absorbable suture materials for closure of jejunal enterotomy incisions in healthy dogs

Summary

The macroscopic and histological appearance of jejunal antimesenteric incisions approximated with two different absorbable suture materials (monofilament versus multifilament) and three closure techniques (appositional single layer, crushing single layer, and double layer) were compared in healthy dogs at 14 or 28 days, postoperatively. No significant differences between the two suture materials were observed for most of the macroscopic or histological variables. However, the monofilament suture material caused significantly more fibrous tissue reaction in the muscular layer of the jejunum than did the multifilament suture material. Of the three enterotomy closure techniques used in this study, the appositional single‐layer method proved to be the best. The double‐layer closure method caused a significant decrease in the incisional circumference, the relative circumference, and volume of the jejunum, and a significant increase in jejunal wall thickness. Our findings suggest that canine jejunal enterotomy incisions can be closed using an appositional suture pattern with relatively rapidly absorbable monofilament suture material. The use of double‐layer suture patterns for closure of jejunal enterotomy incisions should be avoided because the size of the intestinal lumen may be reduced.     

Clinical efficacy and cardiorespiratory effects of alfaxalone, or diazepam/fentanyl for induction of anaesthesia in dogs that are a poor anaesthetic risk

ObjectiveTo evaluate the clinical efficacy and cardiorespiratory effects of alfaxalone as an anaesthetic induction agent in dogs with moderate to severe systemic disease.Study designRandomized prospective clinical study.AnimalsForty dogs of physical status ASA III-V referred for various surgical procedures.MethodsDogs were pre-medicated with intramuscular methadone (0.2 mg kg^?1) and allocated randomly to one of two treatment groups for induction of anaesthesia: alfaxalone (ALF) 1–2 mg kg^?1 administered intravenously (IV) over 60 seconds or fentanyl 5 μg kg^?1 with diazepam 0.2 mg kg^?1± propofol 1–2 mg kg^?1 (FDP) IV to allow endotracheal intubation. Anaesthesia was maintained with isoflurane in oxygen and fentanyl infusion following both treatments. All dogs were mechanically ventilated to maintain normocapnia. Systolic blood pressure (SAP) was measured by Doppler ultrasound before and immediately after anaesthetic induction, but before isoflurane administration. Parameters recorded every 5 minutes throughout subsequent anaesthesia were heart and respiratory rates, end-tidal partial pressure of carbon dioxide and isoflurane, oxygen saturation of haemoglobin and invasive systolic, diastolic and mean arterial blood pressure. Quality of anaesthetic induction and recovery were recorded. Continuous variables were assessed for normality and analyzed with the Mann Whitney U test. Repeated measures were log transformed and analyzed with repeated measures anova (p < 0.05).ResultsTreatment groups were similar for continuous and categorical data. Anaesthetic induction quality was good following both treatments. Pre-induction and post-induction systolic blood pressure did not differ between treatments and there was no significant change after induction. The parameters measured throughout the subsequent anaesthetic procedures did not differ between treatments. Quality of recovery was very, quite or moderately smooth.Conclusions and clinical relevanceInduction of anaesthesia with alfaxalone resulted in similar cardiorespiratory effects when compared to the fentanyl-diazepam-propofol combination and is a clinically acceptable induction agent in sick dogs.     

Evaluation of tricuspid annular plane systolic excursion measured by two-dimensional echocardiography in healthy dogs: repeatability,reference intervals,and comparison with M-mode assessment

Introduction

We sought to determine the feasibility, measurement variability, and within-day repeatability of tricuspid annular plane systolic excursion (TAPSE) measured by two-dimensional echocardiography (2D TAPSE), generate reference intervals for 2D TAPSE, assess agreement and correlation between 2D TAPSE and the conventional TAPSE measured by M-mode echocardiography (MM TAPSE), and to assess the ability of 2D TAPSE to track a drug-induced decrease in right ventricular (RV) function compared with MM TAPSE.

Two techniques for supplementing interlocking nail repair of fractures of the humerus, femur, and tibia: results in 12 dogs and cats

OBJECTIVE: To describe 2 devices for improving stabilization of inadequately stabilized interlocking nail (ILN) repairs of the humerus, tibia, and femur in dogs and cats. STUDY DESIGN: Prospective study. ANIMALS: Twelve client-owned dogs and cats. METHODS: Two devices to further stabilize ILN repair of inadequately stabilized diaphyseal fractures were developed. Device 1 was an axial extension for the ILN that was connected to a conventional type I external skeletal fixator (ESF) with a short connecting bar. Device 2 had hybrid ILN bolt/ESF pins that were used to lock the ILN and serve as the pins for a type I ESF. Devices were used at the initial surgery when the stability of ILN repair was considered inadequate based on palpable fracture segment movement, insufficient medullary canal filling of the ILN at the fracture site, or when the ILN was used in a buttress mode. Outcome was obtained by recheck examinations, radiography, and telephone interview. RESULTS: Device 1 was applicable to fractures of the humerus and femur, but was not used for fractures of the tibia because the ILN extension would have interfered with the stifle. No gross loosening of the ILN/ESF extension connection to the ILN occurred. Device 2 was easily placed and used in the humerus, femur, and tibia. Device 2 allowed removal of the ILN interlock to one or both main fracture segments non-invasively. Clinically, both devices added stability compared with ILN repair alone. Both devices facilitated controlled destabilization of the fracture repair as healing progressed. Complications of pin tract infection, and premature hybrid bolt/ESF pin loosening resulting in premature ESF removal each occurred in 1 patient. Four of 28 hybrid ILN/ESF pins were grossly loose at 4- or 6-week postoperative recheck examinations. Outcomes were excellent (9), good (1), fair (1), and poor (1). CONCLUSIONS: Inadequately stabilized ILN repair of fractures can be stabilized by use of either device, both of which also permit controlled destabilization of the repair during healing. Device 2 can be used when non-invasive removal of the ILN interlock is desired during healing. CLINICAL RELEVANCE: These 2 devices should be considered as alternative methods for stabilization of inadequately stabilized ILN repairs in dogs and cats, or when controlled destabilization of an ILN fracture repair is desired.     

Nippostrongylus brasiliensis infection in the rat: 2. A comparison of production of antibodies in bile and serum by baby rats,lactating rats and adult rats

Production of bile lgA and serum lgG antibody responses against whole worm antigens and worm metabolic product antigens occurred by 7 to 14 days after a single infection with $\text{N. brasiliensis}$ and was similar in adult rats, lactating rats and baby rats. Production of bile lgA antibodies against phosphorylcholine was greater in adult rats, 7 days after infection, than in baby rats or lactating rats. lgA antibody production by baby rats may be affected by immaturity of the intestinal lgA system, and diversion to the mammary gland of lgA-plasma cell precursors may occur in lactating rats. Such effects may be involved in the failure of baby rats and lactating rats to expel parasites by day 21 at which time in adult rats expulsion was complete.     

A comparison of cardiopulmonary function,recovery quality,and total dosages required for induction and total intravenous anesthesia with propofol versus a propofol‐ketamine combination in healthy Beagle dogs

ObjectiveTo compare cardiopulmonary function, recovery quality, and total dosages required for induction and 60 minutes of total intravenous anesthesia (TIVA) with propofol (P) or a 1:1 mg mL⁻¹ combination of propofol and ketamine (KP).Study designRandomized crossover study.AnimalsTen female Beagles weighing 9.4 ± 1.8 kg.MethodsDogs were randomized for administration of P or KP in a 1:1 mg mL⁻¹ ratio for induction and maintenance of TIVA. Baseline temperature, pulse, respiratory rate (f_R), noninvasive mean blood pressure (MAP), and hemoglobin oxygen saturation (SpO₂) were recorded. Dogs were intubated and spontaneously breathed room air. Heart rate (HR), f_R, MAP, SpO₂, end tidal carbon dioxide tension (Pe’CO₂), temperature, and salivation score were recorded every 5 minutes. Arterial blood gas analysis was performed at 10, 30, and 60 minutes, and after recovery. At 60 minutes the infusion was discontinued and total drug administered, time to extubation, and recovery score were recorded. The other treatment was performed 1 week later.ResultsKP required significantly less propofol for induction (4.0 ± 1.0 mg kg⁻¹ KP versus 5.3 ±1.1 mg kg⁻¹ P, p = 0.0285) and maintenance (0.3 ± 0.1 mg kg⁻¹ minute⁻¹ KP versus 0.6 ±0.1 mg kg⁻¹ minute⁻¹ P, p = 0.0018). Significantly higher HR occurred with KP. Both P and KP caused significantly lower MAP compared to baseline. MAP was significantly higher with KP at several time points. P had minimal effects on respiratory variables, while KP resulted in significant respiratory depression. There were no significant differences in salivation scores, time to extubation, or recovery scores.Conclusions and clinical relevanceTotal intravenous anesthesia in healthy dogs with ketamine and propofol in a 1:1 mg mL⁻¹ combination resulted in significant propofol dose reduction, higher HR, improved MAP, no difference in recovery quality, but more significant respiratory depression compared to propofol alone.