Comparison of 5 Surgical Techniques for Partial Liver Lobectomy in the Dog for Intraoperative Blood Loss and Surgical Time

Objective: To compare surgical time and intraoperative blood loss for 5 partial liver lobectomy techniques in the dog. Study Design: Experimental in vivo study. Animals: Dogs (n=10). Methods: Five surgical techniques (SurgiTie^?; LigaSure^?; Ultracision^® Harmonic Scalpel [UAS]; Suction+Clip; Suction+thoracoabdominal stapler [TA]) for partial liver lobectomy in dogs were evaluated and compared for total surgical time and intraoperative blood loss. Body weight, activated clotting time (ACT), heart rate, and intraoperative blood pressure (BP) were recorded. Blood loss was determined by adding the weight of the blood soaked sponges during surgery (1 g=1 mL) to the amount of suctioned blood (mL). Surgical time (in seconds) was determined from the start of the lobectomy until cessation of bleeding from the stump. Mean surgical time and mean blood loss for each technique were compared using a Tukey's multiple comparison test. Results: No significant differences were found between dogs for weight, ACT, heart rate, and intraoperative BP. No complications were seen with the SurgiTie^? technique in 9 of 10 cases. There was no significant difference in surgical time between techniques however there was a significant difference for blood loss; the Suction+Clip method had significantly more blood loss than the other techniques. Conclusions: Skeletonization of the lobar vessels before individually clipping them (Suction+Clip) resulted in a higher blood loss than using Suction+TA, UAS, SurgiTie^? or the LigaSure^? device. The SurgiTie^? appears to be an acceptable method for partial liver lobectomy. Clinical Relevance: Although skeletonization and individually clipping the vessels had the highest blood loss, it still was <7.5% of total blood volume. All 5 techniques should be safe for clinical use in small to medium sized dogs up to 26 kg.     

Evaluation of three methods for measurement of hemoglobin and calculated hemoglobin variables with the ADVIA 120(R) and ADVIA 2120(R) systems in goats

The present study investigated 3 methods of hemoglobin (Hb) determination in goats using the ADVIA 120 and ADVIA 2120 systems. Ethylenediaminetetraacetic acid anticoagulated caprine blood samples (n = 40 goats) were subjected to Hb determination via the cyanmethemoglobin methods in both instruments and a novel, cyanide-free, colorimetric method with the ADVIA 2120. Statistical analysis of the data included a linear regression, Passing-Bablok regression, and Bland-Altman diagram. Colorimetric Hb results determined with both analyzers had excellent correlation (r = 0.98); however, a mean proportional bias of -19.1% was present in comparison to the reference method. There also was excellent agreement between cellular Hb concentrations when measured with both analyzers (r = 0.96), and the constant bias was close to zero. However, imprecision was higher compared to colorimetric methods. Excellent to fair agreement was evident for all calculated erythrocyte and Hb variables. Because of the excellent correlation between the ADVIA 120 and ADVIA 2120, the cyanide-free method of Hb determination could be used with caprine blood specimens; however, the proportional bias must be considered.     

Buccal mucosa bleeding times of healthy dogs and of dogs in various pathologic states, including thrombocytopenia, uremia, and von Willebrand's disease

The buccal mucosa bleeding time (BMBT; duration of hemorrhage from standardized cuts made with a spring-loaded disposable device in the mucosal surface of the upper lip) was used to evaluate the hemostatic competence of dogs. The mean (+/- SD) BMBT for 34 healthy dogs was 2.62 +/- 0.49 minutes. The BMBT of healthy dogs anesthetized with halothane or tranquilized with xylazine were not significantly different from the BMBT of healthy dogs evaluated without chemical restraint. The BMBT was significantly (P less than 0.01) prolonged 21 hours after aspirin (10 mg/kg of body weight) was administered orally to 10 healthy dogs; however, the mean aspirin-induced increase in BMBT was only 0.40 minutes. The BMBT of 28 of 30 dogs with various diseases not traditionally associated with hemostatic deficiencies were near or within the range of BMBT for healthy dogs; however, 2 dogs had BMBT of greater than 8 minutes. In contrast, BMBT were prolonged in most dogs with diseases known to induce deficient primary hemostasis; the 3 dogs with thrombocytopenia (less than or equal to 20,000 platelets/microliter), the 7 Doberman Pinschers with von Willebrand's disease (von Willebrand factor antigen; less than or equal to 18 U/dl), and 5 of the 6 dogs with severe azotemia (serum urea nitrogen; greater than or equal to 124 mg/dl) had prolonged BMBT. The BMBT of 16 dogs were determined immediately before they were subjected to various surgical procedures, and the severity of the hemorrhage encountered during these procedures was subjectively evaluated; the amount of hemorrhage from 12 of the 16 dogs was considered to be appropriate for the corresponding surgical procedures, but the remaining 4 dogs bled excessively during surgery.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of meloxicam on the haemostatic profile of dogs undergoing orthopaedic surgery

The buccal mucosal bleeding time (BMBT), prothrombin time (PT), activated partial thromboplastin time (APTT) and intraoperative bleeding score (IBS) of 38 dogs that underwent orthopaedic surgical procedures and received meloxicam orally and/or parenterally were measured. Fourteen of the dogs (group A) received a single subcutaneous dose of 0.2 mg/kg meloxicam at premedication, 18 dogs (group B) received 0.1 mg/kg meloxicam orally daily for five days followed by a single subcutaneous dose of 0.2 mg/kg meloxicam preoperatively, and six dogs (group C) received 0.5 ml of normal saline subcutaneously at premedication. No statistically significant differences among the groups were detected in relation to the mean (SD) values of BMBT, PT and IBS before and after the surgery, or in the values of APTT in group A. In group B there was a small but significant increase in APTT after the surgery, but all the measurements were within the normal range for dogs.     

Analgesic comparison of meloxicam or ketoprofen for orthopedic surgery in dogs

OBJECTIVE: To compare the safety and efficacy of 2 analgesic protocols (preoperative meloxicam or intraoperative ketoprofen administration) during the first 24 hours after orthopedic surgery in dogs. STUDY DESIGN: Double-blind, prospective randomized clinical trial. ANIMALS: Sixty client-owned dogs. METHODS: Dogs with surgical orthopedic disorders were randomly separated into 2 groups: 30 dogs were administered 0.2 mg/kg meloxicam intravenously (IV) immediately before induction and 30 dogs were administered 2 mg/kg ketoprofen IV, 30 minutes before the end of surgery. Pain was assessed with a visual analog scale (VAS) and a cumulative pain score (CPS) preoperatively and at 30 minutes, 1, 2, 3, 4, 6, 8, and 24 hours after extubation. Selected serum biochemical variables were measured before and 24 hours after surgery and, buccal mucosal bleeding time (BMBT) and whole blood clotting time (WBCT) were measured before and 8 hours after surgery. Dogs were anesthetized with propofol and maintained on halothane in oxygen. Any complications were documented for 7 days after surgery. Results were compared between the 2 groups for significant differences in VAS scores (2-sample t-test) and in CPS (Wilcoxon's 2-sample test). Moreover, results were analyzed for significant differences in area under the curve (AUC) for VAS (2-sample t-test) and CPS (Wilcoxon's 2-sample test) among groups. To assess the effects of treatments on biochemical and coagulation functions, pre- and postoperative mean values of BMBT and WBCT were compared within both treatment groups (paired t-tests) and between both groups (2-sample t-test). RESULTS: No significant differences in pain response or coagulation were found between meloxicam- and ketoprofen-treated dogs. In both groups, alkaline phosphatase and alanine aminotransferase concentrations were significantly increased compared with baseline. No serious complications occurred. CONCLUSIONS: Preoperative administration of meloxicam is a safe and effective method of controlling postoperative pain for up to 24 hours in dogs undergoing orthopedic surgery. CLINICAL RELEVANCE: Analgesia after administration of preoperative meloxicam was comparable with administration of ketoprofen at the end of the surgery.     

Comparison of a method using the HemoCue near patient testing device with a standard method of haemorrhage estimation in dogs undergoing spinal surgery

Objective  To compare an estimate of blood loss obtained using measurements from the Hemocue photometer with a standard estimate in dogs undergoing spinal surgery.
Study design  Prospective clinical study.
Animals  Twenty-nine client-owned dogs.
Methods  During surgery, blood and all lavage fluids were collected in the suction container and on to swabs. To prevent blood clot formation in the suction container, 10 mL citrate-phosphate dextrose adenine (CPDA) was added. At the end of the procedure, the total volume in the suction container was recorded. It was shaken to ensure uniformity and a 5 mL sample tested with the HemoCue photometer. Blood loss in the suction container was calculated as follows: Blood in suction (mL) = volume in bottle (mL) × [suction haemoglobin (Hb) concentration (g dL⁻¹)/pre-operative Hb concentration (g dL⁻¹)]. This volume was added to the estimated volume of blood on the swabs (weight of soaked swabs minus that of dry swabs) to provide the Hemocue estimate of total blood loss. A standard haemorrhage estimate was performed using the volume of fluid in the suction container at the end of surgery in excess of the total volume of lavage fluid available, minus 10 mL CPDA. This volume was added to the estimated volume of blood on the swabs to provide the standard estimate of total blood loss. Data were analyzed with a paired t -test. Retrospective power calculations demonstrated an 80% power to detect a mean difference of 25 mL between the two methods with a level of significance of 0.05.
Results  There was no significant difference in calculated blood loss between the two methods ( p  = 0.8, mean difference: −2 mL, 95% CI: −20 to 16 mL).
Conclusions and clinical relevance  The HemoCue may be used to help estimate blood loss in dogs undergoing spinal surgery.     

The use of haemostatic gelatin sponges in veterinary surgery

Objectives : To describe the use of absorbable gelatin sponges as haemostatic implants in clinical veterinary surgical cases and to document any related postoperative complications. Methods : Practice databases were searched for the product names “Gelfoam” and “Spongostan”. Patient records were retrieved and data regarding patient signalment, surgical procedure, National Resource Council (NRC) wound classification, source of haemorrhage, pre‐ and postoperative body temperature, postoperative complications, time to discharge and details of any postoperative imaging were recorded and reviewed. Follow‐up information was obtained by repeat clinical examination or telephone interview with either the owner or referring veterinary surgeon. Cases with incomplete surgical records or those which were not recovered from anaesthesia were excluded from the analysis. Results : Fifty cases (44 dogs and 6 cats) satisfied the inclusion criteria. Satisfactory haemostasis was achieved in 49 cases with one case requiring reoperation during which a second gelatin sponge was used. There were no detected hypersensitivity responses or confirmed postoperative complications relating to the use of gelatin sponges during the follow‐up period (median 13 months). Clinical Significance : This is the first review of the use of gelatin sponges in clinical veterinary surgery and suggests that gelatin sponges are safe to use in cats and dogs.     

Fluid therapy and newer blood products.

A fluid therapy plan for a patient is developed prior to surgery and is designed to meet each patient's needs. The volume and type of fluid are dependent on the patient's physical status; the acid-base, fluid, and electrolyte status; the surgical procedure; and the expected losses occurring during the procedure. No one fluid regimen is ideal for all patients. All fluid regimens must be continually re-evaluated. A brief minor surgical procedure in a healthy surgical candidate requires little or no fluid administration. In cases of more extensive surgical procedures involving invasion of the abdomen or chest as well as in cases with trauma and major blood loss, much more volume and a specific balanced replacement fluid are required. Depending on the severity of the surgical case, administration rates of 5 to 15 mL/kg/h or greater of crystalloid may be required to maintain perfusion. These rates are merely guidelines, and resuscitation should continue until the desired end point is reached. Balanced replacement fluids may be used to replace blood loss at a ratio of 3:1 and are added to maintenance and replacement requirements. Blood loss of 20% to 25% of the calculated blood volume or hematocrit values less than 20% are indications for colloids or blood replacement at a ratio of 1:1. The optimal fluid therapy regimen for a patient may involve a combination of crystalloids as well as natural and synthetic colloids, using each type of fluid to obtain and maintain perfusion and oxygenation to the tissues.     

The levels of selenium and glutathione peroxidase activity in blood of sheep, cows and pigs.

The levels of selenium (Se) and glutathione peroxidase (GPX) in the blood of sheep, cows and pigs under farm conditions were examined. Sheep appear to form two distinct groups, namely high Se and GPX and low Se and GPX. The high group gave ranges of 133-249 ng/ml and 77-179 iu/g Hb for blood Se and GPX respectively, while the low group showed levels of 21-67 ng/ml and 2-20 iu/g Hb. Overall sheep blood showed a high correlation between Se and GPX (r = 0-92, P less than 0-001). Cow bloods formed one group, all having low Se and GPX levels except for a single outlier. Omitting this animal, the overall ranges were 9-72 ng/ml and 6-36 iu/g Hb for Se and GPX respectively. Blood Se and GPX activity were significantly correlated (r = 0-59, P less than 0-001). Pigs formed a single group also, with the difference that while their blood Se was high, the corresponding blood GPX activities were relatively low. Overall ranges were 93-193 ng/ml and 17-69 iu/g Hb for Se and GPX respectively. Correlation between blood Se level and GPX activity in this species was not significant (r = 0-27, P more than 0-1).     

Effects of DDAVP administrated subcutaneously in dogs with aspirin-induced platelet dysfunction and hemostatic impairment due to chronic liver diseases

To evaluate the hemostatic effects of desmopressin (DDAVP) in dogs with aspirin-induced platelet dysfunction and hemostatic impairment in chronic liver diseases, 3 microg/kg DDAVP was administrated subcutaneously. In aspirin-induced platelet dysfunction dogs (n=5), prolonged BMBT (buccal mucosal bleeding time) was shortened significantly after DDAVP injection (2.2 +/- 1.2 min, P<0.05). In dogs with chronic liver diseases (n=4), activated partial thromboplastin time (APTT) tended to shorten by 0.9 to 3.0 sec, and prolonged BMBT was shortened in two cases for 4.2 and 1.7 min after DDAVP injection. Therefore, the present results indicated that DDAVP shortened the prolonged BMBT in dogs with aspirin-induced platelet dysfunction and chronic liver disease. DDAVP might be helpful in hemostasis under invasive procedures such as biopsy or surgery for dogs with hemostatic impairment.     

Increased cartilage oligomeric matrix protein concentrations in equine digital flexor tendon sheath synovial fluid predicts intrathecal tendon damage

Objectives: To evaluate digital flexor tendon sheath (DFTS) synovial fluid cartilage oligomeric matrix protein (COMP) concentrations as a molecular marker for intrathecal pathology. Study Design: Case control study. Animals: Horses (n=46) with DFTS tenosynovitis; 23 fresh cadaver horses. Methods: DFTS synovial fluid samples were collected from clinical cases with noninfected DFTS tenosynovitis and from control DFTS. Clinical and surgical findings were recorded, and dissection of control limbs was performed to confirm the DFTS to be grossly normal. Synovial fluid COMP was quantified using a homologous competitive inhibition ELISA. Results: Abnormalities were identified tenoscopically: intrathecal tendon/ligament tearing was identified in 37 cases and 9 had other lesions. In control horses, synovial fluid COMP was higher in younger horses. Clinical cases with intrathecal tendon/ligament tearing had higher synovial fluid COMP than either clinical cases with other lesions, or controls. In horses ≥5 years old, the sensitivity and specificity of the assay was high for diagnosing intrathecal tendon/ligament tearing. Conclusions: COMP concentrations in DFTS synovial fluid were significantly greater than those in normal horses with noninfected tenosynovitis caused by intrathecal tendon/ligament tearing, but not by other lesions.     

Assessment of glycosaminoglycan concentration in equine synovial fluid as a marker of joint disease.

A modification of a colorimetric assay was used to determine synovial fluid total and individual sulphated-glycosaminoglycan concentration in various clinical presentations of joint disease in horses. Concentrations of synovial fluid and serum sulphated-glycosaminoglycan (GAG) were measured by the 1,9-dimethylmethylene blue (DMMB) dye assay in normal horses (n = 49), horses with acute (n = 26) or chronic (n = 27) joint disease (defined by clinical, radiographic, and clinicopathological parameters), and horses with cartilaginous lesions at diagnostic arthroscopy, but with normal radiographs and synovial fluid (n = 9). Horses with acute joint disease were subdivided into moderate acute (n = 21) and severe acute (n = 5) joint disease on the basis of synovial fluid analysis and clinical examination. Horses with chronic joint disease were subdivided into mild chronic (n = 9), moderate chronic (n = 10), and severe chronic (n = 8) joint disease on the basis of synovial fluid analysis, clinical examination, and radiographic findings. The concentrations of chondroitin sulphate (CS) and keratan sulphate (KS) were analyzed in each sample following sequential enzymatic digestion of the sample with chondroitinase or keratanase. In addition, the concentration of hyaluronate (HA) in each sample was determined by a colorimetric assay following digestion of the sample with microbial hyaluronidase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Analysis of feline,canine and equine hemograms using the QBC VetAutoread

Blood samples form 120 consecutive clinical cases (40 cats, 40 dogs and 40 horses) were analyzed on the QBC VetAutoread analyzer and the results compared with those obtained by a Baker 9000 electronic resistance cell counter and a 100-cell manual differential leukocyte (WBC) count. Packed cell volume (PCV), hemoglobin (Hb) concentration, mean cell hemoglobin concentration (MCHC), and platelet, total WBC, granulocytes, and lymphocyte plus monocyte (L+M) counts were determined. Indistinct separation of red blood cell and granulocytes layers on the QBC VetAutoread was observed in samples from five cats (12.5%), two dogs (5%), and one horse. Significantly different (P=0.002) median values for the two methods were obtained for PCV, Hb concentration, MCHC and platelet count in cats; PCV, MCHC, WBC, count and granulocytes count in dogs; and PCV, Hb concentration, MCHC and WBC, granulocytes and platelet counts in horses. Results from the QBC VetAutoread should not be interpreted using reference ranges established using other equipment. Results were abnormal on a limited number of samples; however, when correlation coefficients were low, marked discrepancy existed between values within as well as outside of reference ranges. Spearman rank correlation coefficients were excellent (r=0.93) for PCV and Hb concentration in dogs, and Hb concentration and WBC count in horses. Correlation was good (r=0.80-0.92) for PCV and Hb concentration in cats, WBC count in dogs, and PCV, granulocytes count and platelet count in horses. For remaining parameters, correlation was fair to poor (r=0.79). Acceptable correlations (r>0.80) were achieved between the two test systems for all equine values except MCHC and L+M count, but only for PCV and HB concentration in feline and canine blood samples.     

Comparison of lipase activity in peritoneal fluid of dogs with different pathologies--a complementary diagnostic tool in acute pancreatitis?

A clinical diagnosis of acute pancreatitis is often difficult to obtain. Histopathology remains the gold standard, whereas clinical signs, diagnostic imaging and laboratory testing, even in combination, may be insufficient. In a prospective study, lipase activity in ascitic fluid of various aetiologies was determined in 44 dogs in order to investigate its performance in cases of acute pancreatitis. Data of simultaneously determined blood lipase activities were available in 27 dogs. Lipase activity was measured by a colorimetric assay. A complete peritoneal fluid analysis was performed. Dogs were divided into four groups, according to their final diagnosis: acute pancreatitis (A), abdominal trauma (B), abdominal neoplasia (C) and others (hepatic or cardiac diseases) (D). Dogs with acute pancreatitis had a significantly higher peritoneal lipase activity than those in other groups (P < or = 0.024), while no significant difference was found between the other groups (P > or = 0.734). Blood lipase activity as well as protein content and total cell count of the ascitic fluid did not show any significant difference between groups. Data show that determination of lipase activity in dogs that develop ascites may be useful in complementing the diagnosis of acute pancreatitis.     

Evaluation of a fluorometric method for the quantitative assay of fecal hemoglobin in the dog

Feces from 27 dogs were evaluated to establish baseline fecal hemoglobin (Hb) concentrations when the dogs were fed diets containing varying amounts of Hb. Fecal Hb concentration was measured, using a quantitative fluorometric assay that was based on the conversion of nonfluorescing Hb heme to fluorescing porphyrins. Mean fecal Hb concentration was 0.31 mg/g of feces when the dogs were fed a dry diet low in Hb. This corresponded to a daily fecal blood loss of 0.043 ml/kg of body weight. The fecal Hb concentration was directly related to the dietary Hb concentration. The average recovery of orally ingested blood was 41% in 4 dogs. This fluorometric assay quantitatively detected small amounts of gastrointestinal bleeding over a wide range of fecal Hb concentrations for which feces appeared normal. Results of this study establish dietary conditions necessary for quantitative evaluation of experimental and clinical gastrointestinal bleeding.     

Hemostatic defects associated with two infusion rates of dextran 70 in dogs.

We investigated changes in hemostatic function after infusion of 6% dextran 70 (high molecular weight dextran) at 2 rates. Six healthy dogs underwent 3 regimens: 20 ml of dextran/kg of body weight administered in 1 hour (trial A), 20 ml of dextran/kg administered in 30 minutes (trial B), and 0.9% sodium chloride solution as a control administered over 1 hour to achieve hemodilution equivalent to that for 20 ml of dextran/kg (trial C). Before and at 2, 4, 8, and 24 hours after the start of trials A and B, we measured PCV, total solids (TS) concentration, amount of von Willebrand factor antigen (vWf:Ag), factor VIII coagulant activity (VIII:C), prothrombin time, activated partial thromboplastin time (APTT), platelet retention in a glass bead column, and buccal mucosa bleeding time (BMBT). Values were not obtained at 8 and 24 hours for trial C. Saline-induced changes in hemostasis were significant (P less than 0.05) from baseline throughout the sample collection period. Significant differences (P less than 0.05) between trial A and control were observed for vWf:Ag, VIII:C, BMBT, APTT, TS, and PCV values at 2 hours, and for VIII:C at 4 hours. Significant differences (P less than 0.05) between trial B and control were observed for APTT, TS, and PCV values at 2 hours, and for vWf:Ag, VIII:C, BMBT, APTT, TS, and PCV values at 4 hours. During trials A and B, mean values of analytes infrequently deviated from reference intervals, and clinical signs of bleeding were not observed in any dog.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of desmopressin acetate administration on primary hemostasis in Doberman Pinschers with type-1 von Willebrand disease as assessed by a point-of-care instrument

OBJECTIVE: To evaluate primary hemostasis following administration of desmopressin acetate (DDAVP) to Doberman Pinschers with type-1 von Willebrand disease (vWD). ANIMALS: 16 nonanemic Doberman Pinschers with type-1 vWD. PROCEDURE: Closure time (CT), defined as time required for occlusion of an aperture by a platelet plug assessed within the point-of-care instrument, plasma von Willebrand factor (vWF) concentration, and buccal mucosal bleeding time (BMBT) were determined before and 1 hour after administration of DDAVP (1 microg/kg, SC). RESULTS: Baseline closure times measured with adenosine diphosphate ([ADP-CT], 108 to > 300 seconds; reference range, 52 to 86 seconds) and epinephrine ([EPI-CT], 285 to > 300 seconds; 97 to 225 seconds) as platelet agonists were prolonged in all dogs. Following DDAVP administration, ADP-CT (59 to 186 seconds) was significantly shortened from baseline, but there was no decrease in EPI-CT. Although mean plasma vWF concentration increased significantly after DDAVP administration, only 1 dog had an increase of > 35 U/dL. There was no correlation between increase in plasma vWF concentration and shortening of the ADP-CT. Baseline BMBT was prolonged in 12 of 14 dogs, with significant shortening of BMBT after DDAVP administration in 6 of 7 dogs. In vitro replacement of vWF-deficient plasma with plasma from an unaffected dog shortened the ADP-CT whereas in vitro addition of DDAVP had no effect. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of DDAVP to Doberman Pinschers with type-1 vWD resulted in improved hemostatic function, as assessed by the point-of-care instrument and shortening of BMBT, despite minimal increase in plasma vWF concentration.     

Assessment of fresh extended equine semen samples using spectrophotometry and resazurin

The purpose of this study was to determine if spectrophotometric assessment of resazurin dye in fresh extended equine semen samples was associated with spermatozoal parameters.This technique could be beneficial to veterinarians and horse producers for evaluating semen samples prior to artificially inseminating a mare. The reducible dye resazurin (blue color) is reduced via an oxidation-reduction reaction in the presence of metabolically active spermatozoa to resorufin (pink color), and upon further reduction to dihydroresorufin (colorless). Sixty semen samples were collected from six stallions (5 Quarter Horse and 1 Arabian) using a Missouri style artificial vagina. Sample aliquots were diluted using a 1:30 (semen: extender) ratio with a non-fat dry skim milk (NFDSM) glucose extender T. The diluted sample was then assessed microscopically at 250x to determine concentration, the number of motile, and progressively motile spermatozoa/mL. The remainder of the sample was diluted at a 1:1 (semen: extender) ratio prior to dye incubation and spectrophotometric analysis. The resazurin dye (50 μL from a 0.338 mM solution) was added to 4 (2 mL) aliquots of extended sample, thoroughly mixed, and incubated at 37°C. Butyl alcohol (4.8 mL) was added at five-minute increments (0,5, 10, and 15 minutes) to stop spermatozoal metabolism and draw the color out of the sample. Each aliquot was then vortexed prior to centrifugation at 700xg to extract the butanol color layer. Spectrophotometric absorbance values (615 nm) of the butanol color layer were recorded. Relationships between spectrophotometric absorbance values and spermatozoal parameters were assessed using correlation analyses on square root transformed data. At the 0 minute incubation time there were no associations between spermatozoal parameters and spectrophotometric absorbance values. However, at the five minute incubation time the spectrophotometric absorbance values were negatively correlated with concentration (r=−0.31; P=0.02), number of motile (r=−0.27; P=0.04) and progressively motile (r=−0.30; P=0.02) spermatozoa/mL. At the 10 minute incubation time negative correlations were observed between the spectrophotometric absorbance values and concentration (r=−0.48; P=0.0001), number of motile (r=−0.45; P=0.0004) and progressively motile (r=−0.46; P=0.0002) spermatozoa/mL. At the 15 minute incubation time negative correlations were also found between spectrophotometric absorbance values and concentration (r=−0.52; P=0.0001), number of motile (r=−0.50; P=0.0001) and progressively motile (r=−0.52; P=0.0001) spermatozoa/mL. Spectrophotometric absorbance values were associated with spermatozoal parameters at the 5, 10, and 15 minute incubation times.     

Evaluation of three methods for measurement of hemoglobin and calculated hemoglobin parameters with the ADVIA 2120 and ADVIA 120 in dogs, cats, and horses

BACKGROUND: Besides flow cytometric detection of cellular hemoglobin (HGB) concentration, the ADVIA 2120 uses a novel cyanide-free colorimetric method to determine extracellular total HGB concentration. In human samples, the results are equivalent to those of the cyanmethemoglobin method on the ADVIA 120. Cyanide-free HGB measurement has not been evaluated in animal samples. OBJECTIVES: The aim of this prospective study was to compare the 3 methods of HGB analysis on the ADVIA 2120 and ADVIA 120 in blood samples from dogs, cats, and horses. METHODS: Consecutive fresh K(3)EDTA blood samples from 119 dogs, 113 cats, and 151 horses submitted to the Central Laboratory, Department of Veterinary Clinical Sciences, Justus-Liebig University Giessen, were included. A CBC was performed on each sample using the ADVIA 2120 and ADVIA 120. Colorimetric and cellular HGB concentrations and all calculated variables based on HGB measurement were compared using linear regression, Passing Bablok regression, and Bland Altman plots, using the ADVIA 120 as the reference method. RESULTS: In samples from all species, an excellent correlation was found for colorimetric HGB results (r=0.99). HGB measured with the cyanide-free method was overestimated on the ADVIA 2120 compared with the cyanide-based method on the ADVIA 120, with a mean proportional bias of -21.0% (dog), -22.0% (cat), and -19.4% (horse). The correlation of cellular HGB concentration between analyzers was excellent (r=0.99); however, imprecision was higher than for colorimetric methods. Excellent to fair agreement was found for all calculated variables. CONCLUSION: The cyanide-free method of HGB determination is appropriate for use in blood samples from animals, provided the proportional bias is considered.     

Changes in body composition during weight loss in obese client-owned cats: loss of lean tissue mass correlates with overall percentage of weight lost

Obesity is one of the most common medical diseases in cats, but there remains little information on success of weight loss regimes in obese client-owned cats. No information currently exists on body composition changes during weight loss in clinical cases. Twelve obese client-owned cats undertook a weight loss programme incorporating a high-protein low fat diet. Body composition was quantified by dual-energy X-ray absorptiometry, before and after weight loss. Mean (+/-standard deviation) weight loss was 27+/-6.8% of starting weight, and mean rate of weight loss was 0.8+/-0.32% per week. Mean energy allocation during weight loss was 32+/-7.0 kcal/kg target weight. Mean composition of tissue lost was 86:13:1 (fat:lean:bone mineral). The proportion of lean tissue loss was positively associated with overall percentage of weight loss (simple linear regression, r(2)=44.2%, P=0.026). Conventional weight loss programmes produce safe weight loss, but lean tissue loss is an inevitable consequence in cats that lose significant proportions of their starting body weight.