Evaluation of a chromogenic assay to measure the factor Xa inhibitory activity of unfractionated heparin in canine plasma

BACKGROUND: Unfractionated heparin (UFH) has a complex pharmacologic profile that necessitates patient monitoring to prevent inadequate anticoagulation or overdosage and hemorrhage. Factor Xa inhibitory assays (to measure anti-Xa activity) are used to adjust UFH dosage and define safe and effective regimens for specific thrombotic disorders in humans. OBJECTIVE: In this study, the accuracy, linearity, and clinical utility of a chromogenic assay were assessed for monitoring UFH anti-Xa activity in canine plasma samples. METHODS: A commercial assay (Rotachrom Heparin, Diagnostica Stago, Parsippany, NJ, USA) was used to measure anti-Xa activity in canine plasma samples spiked with different concentrations of UFH. Background absorbance and assay linearity were compared for canine and human plasmas. Percentage recovery of UFH anti-Xa activity and intra- and interassay imprecisions were investigated by multiple measurements of canine plasma to which known amounts of UFH were added. The spiked plasma samples also were used to determine the heparin sensitivity of an activated partial thromboplastin time (aPTT) test. RESULTS: Canine plasma samples were assayed at a higher dilution than were human plasma samples (3:8 versus 4:8) to eliminate higher background anti-Xa activity in canine plasma. Using this modification, the recovery of anti-Xa activity in canine plasma was linear (R2 > .9) at concentrations of 0 - 0.75 U/mL UFH. Intra- and interassay imprecisions for plasma samples containing 0.5 U/mL UFH were <10%, whereas samples containing 0.25 U/mL UFH had imprecisions of 13% and 24%, respectively. The anti-Xa activity range of 0.5 - 0.75 U/mL caused prolongation of aPTTs to 1.5 - 2.5 times the assay mean. CONCLUSION: Plasma anti-Xa activity of dogs treated with UFH can be accurately monitored using this commercially available chromogenic assay.     

Pharmacokinetics and pharmacodynamics of a therapeutic dose of unfractionated heparin (200 U/kg) administered subcutaneously or intravenously to healthy dogs

BACKGROUND: Heparin treatment has been recommended for dogs in hypercoagulable states such as disseminated intravascular coagulation, however, potential benefits have to be balanced against the bleeding risk if overdosage occurs. A better understanding of the pharmacology of heparin and tests to monitor heparin therapy in dogs may help prevent therapeutic hazards. OBJECTIVES: The purpose of this study was to evaluate the effects of 200 U/kg of sodium unfractionated heparin (UFH) on coagulation times in dogs after intravenous (IV) and subcutaneous (SC) administration and to compare these effects with plasma heparin concentrations assessed by its antifactor Xa (aXa) activity. METHODS: 200 U/kg of UFH were administered IV and SC to 5 healthy adult Beagle dogs with a washout period of at least 3 days. Activated partial thromboplastin time (APTT), prothrombin time (PT), and plasma aXa activity were determined in serial blood samples. RESULTS: After IV injection, PT remained unchanged except for a slight increase in 1 dog; APTT was not measurable (>60 seconds) for 45-90 minutes, and then decreased gradually to baseline values between 150 and 240 minutes. High plasma heparin concentrations were observed (maximal concentration = 4.64 +/-1.4 aXa U/mL) and decreased according to a slightly concave-convex pattern on a semilogarithmic curve, but returned to baseline slightly more slowly (t240-t300 minutes) than did APTT. After SC administration, APTT was moderately prolonged (by a ratio of 1.55 +/-0.28 APTT t0, range 1.35-2.01) between 1 and 4 hours after administration. Plasma aXa activity reached a maximum of 0.56 +/-0.20 aXa U/mL (range 0.42-0.9 U/mL) after 132 +/-26.8 minutes; this lasted for 102 +/-26.8 minutes. Prolongation of APTTs of 120-160% corresponded to plasma heparin concentrations of 0.3-0.7 aXa U/mL. CONCLUSIONS: As in humans, the pharmacokinetics of UFH in dogs was nonlinear. Administration of 200 U/kg of UFH SC in healthy dogs resulted in sustained plasma heparin concentrations in accordance with human recommendations for thrombosis treatment or prevention, without excessively increased bleeding risks. In these conditions, APTT can be used as a surrogate to assess plasma heparin concentrations. These findings need to be confirmed in diseased animals.     

Anticoagulant effects of repeated subcutaneous injections of high doses of unfractionated heparin in healthy dogs.

OBJECTIVE: To evaluate s.c. administration of unfractionated heparin (UFH) in accordance with a dosing regimen for high-dose treatment in dogs. ANIMALS: 10 healthy adult Beagles. PROCEDURES: Two groups of dogs (5 dogs/group) were given 6 injections of heparin (500 units of UFH/kg of body weight, s.c.) at intervals of 8 (experiment 1) and 12 (experiment 2) hours. Blood samples were collected before and 4 hours after heparin injections to determine amidolytic heparin activity, activated partial thromboplastin time (APTT), thrombin time, antithrombin activity, platelet count, and Hct. RESULTS: For experiments 1 and 2, mean +/- SD heparin activities before (experiment 1, 1.32 +/- 0.20 U/ml; experiment 2, 0.69 +/- 0.174 U/ml) and 4 hours after the last heparin injection (experiment 1, 1.71 +/- 0.30 U/ml; experiment 2, 1.10 +/- 0.30 U/ml) were higher than values calculated for the regimen used in experiment 1. Results of the investigated thrombin time test system with low thrombin activity were frequently beyond the measurement range, even with UFH activities > or = 0.6 U/ml. Moreover, a severe decrease of antithrombin activity became evident during both experiments (eg, in experiment 2 from 95.6 +/- 4.8 to 59.2 +/- 6.6%). In each treatment group, 2 dogs developed hematomas. CONCLUSIONS AND CLINICAL RELEVANCE: Calculations of the course of heparin activity after a single injection do not result in a reliable dosing regimen for high-dose heparin treatment in dogs. High-dose treatment must be monitored for each dog. Thrombin time measured with low thrombin activity is unsuitable for this purpose.     

The effect of unfractionated heparin on thrombelastographic analysis in healthy dogs

Objective – To determine the effect of single and multiple doses of SQ heparin (200 U/kg) on the thrombelastogram of healthy dogs. Design – Prospective study. Setting – University research facility. Animals – Six random‐source female dogs. Interventions – Baseline parameters, including a CBC with platelet count, prothrombin time, activated partial thromboplastin time (aPTT), and antithrombin were performed. Thrombelastography (TEG) and aPTT were performed hourly for 12 hours after unfractionated heparin dosing (200 U/kg, SQ). Anti‐Xa activity was assayed at 0, 3, 6, and 8 hours. Heparin was then administered every 8 hours for 3 days. The sampling protocol on Day 4 was identical to Day 1. Measurements and Main Results – On Day 1, percentage change from baseline for TEG parameter R, as well as absolute values of K, angle, and maximum amplitude (MA) were evaluated. Statistically significant (P<0.01) prolongation of the R time and a decrease in angle and MA was seen in all dogs by hour 3. R and MA were unmeasurable for most dogs between 3 and 5 hours. All TEG tracings returned to baseline by 12 hours. Day 4 TEG tracings mimicked those on Day 1. Only 1 dog achieved aPTT values outside the reference interval on both days. Anti‐Xa activity levels increased on Day 4 but not on Day 1. Based on post hoc in vitro analysis, prolongation of R time occurred at plasma heparin levels as low as 0.075 U/mL, well below the lower limit of detection of the anti‐Xa activity level assay. Conclusions – Administration of SQ heparin results in progressive changes in the TEG tracing, with maximal change occurring 3–5 hours after dosing. The extensive prolongation of the R time also indicates that TEG may be too sensitive and limits its utility as a monitoring tool for unfractionated heparin therapy.     

Effect of unfractionated and low-molecular-weight heparin on platelet aggregation and in vitro bleeding time in dogs

The aim of the present study was to examine the influence of unfractionated heparin (UFH) and low molecular weight heparin (LMWH) on the function of canine thrombocytes. This was performed by adding different concentrations of UFH or LMWH to platelet rich plasma (PRP) or blood of healthy dogs: 0.2, 0.5, 1, 2, 5, 10, 20, and 50 I.U./ml UFH or Anti-FXaU/ml LMWH, respectively (aggregation induced by thrombin additionally: 0.025, 0.05, and 0.1 I.U./ml UFH or Anti-FXaU/ml LMWH.) Platelet aggregation induced by ADP, collagen or thrombin with the BORN method (n = 11) as well as the in vitro bleeding time using the analyzer PFA-100 (n = 5) were examined. Additionally, a specific test assay for ADP induced platelet aggregation was performed which enabled an individual adjustment of the aggregation maximum at 30-40% in the control measurements (n = 6). The most prominent effect was noted in the platelet aggregation induced by thrombin. The aggregation maximum of the platelet aggregation induced by 1 I.U./ml thrombin (final concentration) was significantly lower in all of the testet UFH concentrations > or = 0.025 I.U./ml UFH in comparison to control measurements. If the aggregation was induced by 10 I.U./ml thrombin a significant reduction of the aggregation maximum was restricted to UFH concentrations > or = 0.5 I.U./ml. The addition of LMWH to canine PRP resulted in a distinct decrease (p < 0.01) of the maximum aggregation induced by 1 I.U./ml thrombin in concentrations > or = 0.2 Anti-FXaU/ml LMWH. A slight decrease of the maximum aggregation induced by collagen was only found for UFH at activities > or = 20 I.U./ml. No significant systematic influence could be demonstrated for LMWH on the aggregation induced by collagen as well as for LMWH and UFH on the platelet aggregation induced by ADP. Capillary in vitro bleeding time (closure time) was prolonged only after adding high concentrations of UFH (> or = 10 I.U./ml) and LMWH (> or = 50 Anti-FXaU/ml) to the sample material. The results document the unimportant influence of therapeutic levels of UFH and LMWH on platelet function in dogs. Therefore, the remarkable inhibition of the aggregation induced by thrombin reflects mainly the antithrombin effect.     

Monitoring unfractionated heparin therapy in dogs by measuring thrombin generation

Background: The calibrated automated thrombogram (CAT), an assay that permits measurement of thrombin generation in plasma, may be useful in studying hemostatic disorders and anticoagulant therapy in animals. Objectives: The aims of the study were to measure thrombin generation in healthy Beagle dogs and to evaluate the potential use of the CAT assay for monitoring therapy with unfractionated heparin (UFH). Methods: Individual platelet‐poor plasma samples and a plasma pool from 20 healthy adult Beagles were prepared. Serial UFH plasma dilutions were used to establish an in vitro heparin‐sensitivity curve. The pharmacodynamic effects of heparin in vivo were evaluated in Beagles using the CAT assay to measure thrombin generation with tissue factor at a concentration of 5 pM for initiation. Results: In healthy Beagles, the range of endogenous thrombin potential (ETP) was 238.7–414.0 nM/min (mean ± SD, 340.4 ± 63.1 nM/min). ETP intra‐assay and interassay variations were 7.1% and 12.9%, respectively. In vitro, a UFH concentration ≥0.4 U/mL resulted in total inhibition of thrombin generation. In vivo, the maximal effect of UFH on ETP was observed at 170 ± 36 minutes (range, 120–210 minutes) and resulted in a decrease in ETP of 38.5 ± 7.8% (range, 26.5–50.3%). In 210–420 minutes, ETP returned to baseline in 5 dogs. Conclusion: Our study demonstrates that thrombin generation can be measured in canine plasma and may be useful in assessing the degree of anticoagulation provided by UFH.     

Clinical Evaluation of a Novel Liquid Formulation of L-Thyroxine for Once Daily Treatment of Dogs with Hypothyroidism

Background: A liquid solution of levothyroxine (L-T4) is available for treatment of canine hypothyroidism.
Hypothesis: Once daily oral administration of a liquid L-T4 solution is effective and safe for controlling hypothyroidism in dogs.
Animals: Thirty-five dogs with naturally occurring hypothyroidism.
Methods: Dogs received L-T4 solution PO once daily at a starting dosage of 20 μg/kg body weight (BW). The dose was adjusted every 4 weeks, based on clinical signs and peak serum total T4 (tT4) concentrations. Target peak serum tT4 and thyroid stimulating hormone (TSH) concentrations, 4–6 hours posttreatment, were 35–95 nmol/L and < 0.68 ng/mL, respectively. Dogs were followed for up to 22 weeks after establishment of the maintenance dose.
Results: Clinical signs of hypothyroidism improved or resolved in 91% of dogs after 4 weeks of L-T4 treatment at 20 μg/kg once daily. The maintenance dose was established in 76, 94, and 100% of dogs after 4, 8, and 12 weeks of treatment, respectively. This was 20 μg L-T4/kg BW for 79% of the dogs, 30 μg/kg BW for 15%, and 10–15 μg/kg BW in the remaining 6%, once daily. Thereafter, median peak tT4 and TSH concentrations were 51 nmol/L and 0.18 ng/mL, respectively, and remained stable during the 22-week follow-up; clinical signs did not recur.
Conclusions and Clinical Importance: All of the hypothyroid dogs had rapid clinical and hormonal responses to supplementation with the PO-administered L-T4 solution. The starting dosage of 20 μg L-T4/kg BW once daily was suitable for 79% of dogs.     

The effect of Hetastarch (670/0.75) in vivo on platelet closure time in the dog

Objective – To evaluate the effect of 6% hydroxyethyl starch (HES) solution in vivo, with an average molecular weight of 670 kDa and degree of substitution of 0.75, on canine platelet function.
Design – Prospective, controlled-experimental study.
Setting – University of California, Davis, Veterinary Medical Teaching Hospital.
Animals – Seven healthy employee-owned dogs.
Interventions – Seven dogs were included in the treatment group. Four of these dogs also served as the control group. Platelet closure time (CT) was measured using a platelet function analyzer and collagen/ADP cartridges. Dogs were given 20 mL/kg of either sodium chloride 0.9% (control group, n =4) or HES (treatment group, n =7) IV over 1 hour. CT was measured before the infusion, and at 1, 3, 5, and 24 hours after the start of the infusion.
Measurements and Main Results – There was a significant change over time from 0 to 24 hours ( P <0.001), a significant difference between groups across time ( P <0.001), and a significant group-by-time interaction ( P =0.007). At 3 hours, mean CT for the treatment group was 122.3±18.1 seconds, which was significantly different ( P <0.001) from the control group (71.0±3.5 s). At 5 hours, mean CT for the treatment group was 142.7±33.9 seconds, which was significantly different ( P =0.001) from the control group (75.0±8.6 s). Mean CT at 24 hours was within the reference interval for both the control and treatment group (66.0±2.9 and 81.8±11.9 s, respectively); however, CT in 3 individual dogs in the treatment group at this time point remained prolonged.
Conclusions – A clinically relevant dose of HES 670/0.75 prolongs CT in dogs for up to 24 hours. This may be due to platelet dysfunction in addition to the effects of hemodilution, and therefore, may increase the risk of bleeding.     

Serum Cardiac Troponin I Concentration in Dogs with Precapillary and Postcapillary Pulmonary Hypertension

Background: Pulmonary hypertension (PH) is a disease condition leading to right-sided cardiac hypertrophy and, eventually, right-sided heart failure. Cardiac troponin I (cTnI) is a circulating biomarker of cardiac damage.
Hypothesis: Myocardial damage can occur in dogs with precapillary and postcapillary PH.
Animals: One hundred and thirty-three dogs were examined: 26 healthy controls, 42 dogs with mitral valve disease (MVD) without PH, 48 dogs with pulmonary hypertension associated with mitral valve disease (PH-MVD), and 17 dogs with precapillary PH.
Methods: Prospective, observational study. Serum cTnI concentration was measured with a commercially available immunoassay and results were compared between groups.
Results: Median cTnI was 0.10 ng/mL (range 0.10–0.17 ng/mL) in healthy dogs. Compared with the healthy population, median serum cTnI concentration was increased in dogs with precapillary PH (0.25 ng/mL; range 0.10–1.9 ng/mL; P < .001) and in dogs with PH-MVD (0.21 ng/mL; range 0.10–2.10 ng/mL; P < .001). Median serum cTnI concentration of dogs with MVD (0.12 ng/mL; range 0.10–1.00 ng/mL) was not significantly different compared with control group and dogs with PH-MVD. In dogs with MVD and PH-MVD, only the subgroup with decompensated PH-MVD had significantly higher cTnI concentration compared with dogs with compensated MVD and PH-MVD. Serum cTnI concentration showed significant modest positive correlations with the calculated pulmonary artery systolic pressure in dogs with PH and some echocardiographic indices in dogs with MVD and PH-MVD.
Conclusions and Clinical Importance: Serum cTnI is high in dogs with either precapillary and postcapillary PH. Myocardial damage in dogs with postcapillary PH is likely the consequence of increased severity of MVD.     

Serum Serotonin Concentrations in Dogs with Degenerative Mitral Valve Disease

Background: Increased serotonin (5HT) signaling has been implicated in valvular disease of humans and animals, including canine degenerative mitral valve disease (DMVD). High circulating 5HT concentration is a potential source of increased signaling, and serum 5HT concentrations have not been previously reported in dogs with DMVD.
Hypothesis: Dogs with DMVD and small breed dogs predisposed to DMVD have higher serum 5HT concentrations than large breed controls.
Animals: Fifty dogs affected with DMVD, 34 dogs predisposed to DMVD but without cardiac murmur or echocardiographic evidence of DMVD, and 36 healthy large breed control dogs.
Methods: Prospective analysis. Serum 5HT concentration was measured by an ELISA test.
Results: Median serum 5HT concentration was significantly higher in dogs with DMVD and in dogs predisposed to DMVD as compared with controls (DMVD, 765.5 ng/mL [interquartile range, 561.3–944.4]; predisposed, 774.9 ng/mL [528.3–1,026]; control, 509.8 ng/mL [320.8–708.8]; P = .0001). Subgroup analysis of predisposed dogs indicated significantly higher serum 5HT concentrations in Cavalier King Charles Spaniel (CKCS) dogs than in other breeds (CKCS, 855.0 ng/mL [635.8–1,088]; non-CKCS, 554.2 ng/mL [380.6–648.4]; P = .0023). Age, platelet count, and platelet morphology were not correlated with 5HT concentration in any group.
Conclusions and Clinical Importance: Dogs with DMVD had significantly higher serum 5HT concentrations when compared with large breed control dogs. Healthy CKCS dogs had significantly higher serum 5HT concentrations than other healthy dogs predisposed to DMVD. Additional investigation into a possible role of 5HT in the pathogenesis of DMVD is warranted.     

Effect of Experimental Hypothyroidism on Glomerular Filtration Rate and Plasma Creatinine Concentration in Dogs

Background: Hypothyroidism affects renal function in a manner opposite the effects of hyperthyroidism.
Objective: To evaluate the effects of experimentally induced hypothyroidism on glomerular filtration rate (GFR) and basal plasma creatinine concentration in dogs.
Animals: Sixteen anestrous, female dogs.
Methods: Hypothyroidism was induced by administration of ¹³¹I in 8 dogs, and 8 healthy euthyroid dogs acted as controls. Exogenous plasma creatinine clearance (an estimate of GFR) was measured in all dogs before (control period) and 43–50 weeks after induction of hypothyroidism (posttreatment period). Other pharmacokinetic parameters of creatinine were also determined.
Results: No significant difference was observed for basal plasma creatinine concentration and creatinine clearance between control and hypothyroid dogs in the control period. In the posttreatment period, mean ± SD creatinine clearance in the hypothyroid group (2.13 ± 0.48 mL/min/kg) was lower ( P < .001) than that of the control group (3.20 ± 0.42 mL/kg/min). Nevertheless, basal plasma creatinine concentrations were not significantly different between the hypothyroid and control groups (0.74 ± 0.18 versus 0.70 ± 0.08 mg/dL, respectively) because endogenous production of creatinine was decreased in hypothyroid dogs (22 ± 3 versus 32 ± 5 mg/kg/d, P =.001).
Conclusion and Clinical Importance: Hypothyroidism causes a substantial decrease in GFR without altering plasma creatinine concentrations, indicating that GFR evaluation is needed to identify renal dysfunction in such patients.     

Prothrombotic and Inflammatory Effects of Intravenous Administration of Human Immunoglobulin G in Dogs

Background: Intravenous administration of human immunoglobulin G (hIVIgG) has been suggested to potentiate thromboembolism in dogs, but supportive scientific reports are lacking.
Objectives: To determine if hIVIgG therapy promotes hypercoagulability and inflammation in dogs.
Animals: Twelve healthy Beagle dogs.
Methods: Prospective, experimental trial. An hIVIgG/saline solution was infused IV at 1 g/kg BW over 8 hours to 6 dogs, and physiological saline was infused to the other 6 dogs. Blood samples were drawn before, during, and after infusion for serial measurement of indicators of coagulation and inflammation. Data were analyzed by 2-way repeated measures analysis of variance.
Results: Dogs administered hIVIgG developed mildly decreased blood platelet concentrations without thrombocytopenia (median, 200 × 10³/μL; range, 150–302 × 10³/μL; P < .01), leukopenia (median, 3.5 × 10³/μL; range, 20–62 × 10³/μL; P < .001), and mildly increased plasma total protein concentrations (median, 6.3 g/dL; range, 5.6–6.7 g/dL; P < .001). Administration of hIVIgG was also associated with increases in fibrin/fibrinogen degradation products in all dogs (either 5 μg/mL or 10 μg/dL), thrombin-antithrombin III complexes (median, 7.2 ng/mL; range, 4.9–14.2 ng/mL; P < .001), and C-reactive protein concentrations (median, 2.5 mg/dL; range, 0.5–4.3 mg/dL; P < .01).
Conclusion and Clinical Importance: Administration of hIVIgG to dogs promotes hypercoagulability and an inflammatory state. This should be further evaluated and considered when using hIVIgG in dogs with IMHA or other prothrombotic conditions.     

Comparison of three techniques for ovarian pedicle hemostasis during laparoscopic-assisted ovariohysterectomy

Objective— To describe the safety, surgical time, and complications associated with 3 techniques for achieving hemostasis during laparoscopic-assisted ovariohysterectomy (LAOVH).
Study Design— Prospective, randomized clinical trial.
Animals— Female dogs ( n =30).
Methods— Dogs were randomly assigned to 1 of 3 methods for achieving ovarian pedicle hemostasis during LAOVH: extracorporeal modified Roeder knot application (suture group), metal clip application using a multifire 10 mm laparoscopic clip applier (clip group), or use of a novel 5 mm bipolar vessel-sealing device (vessel-sealing group). In all dogs a 3 median portal technique was used.
Results— Controlling for the dogs' bodyweights, there was a significant association between surgical time and which method for hemostasis was used. This association was different when comparing the first 5 procedures using each method to the second 5. For a 20 kg dog, the surgical time (95% CI) for the first 5 procedures was 80 (69–91), 68 (57–79), and 33 (21–45) minutes for the suture, clip, and vessel-sealing groups, respectively. For the second 5 procedures surgical time was 71 (60–81), 50 (39–60), and 40 (29–51) minutes. Pedicle hemorrhage occurred in all dogs in the clip group, 3 dogs in the suture group, and none of the dogs in the vessel-sealing group although in all cases was considered hemodynamically inconsequential. All dogs recovered uneventfully.
Conclusions— All methods of hemostasis were safe for pedicle sectioning. A learning curve exists for clip and suture methods.
Clinical Relevance— Use of a vessel-sealing device significantly shortens surgical time and provides excellent hemostasis during LAOVH.     

A prospective, randomized comparison of Oxyglobin (HB-200) and packed red blood cell transfusion for canine babesiosis

Objective – To establish the efficacy of Oxyglobin (HB-200) in canine babesiosis and compare it to standard therapy, packed red blood cell transfusion (pRBCT) with respect to improvements in specific parameters of blood gas, acid-base, blood pressure, and subjective evaluations.
Design – Prospective, randomized, clinical trial.
Setting – Onderstepoort Veterinary Academic Hospital.
Animals – Twelve dogs (8–25 kg) naturally infected with Babesia rossi and a hematocrit of 0.1–0.2 L/L (10–20%).
Interventions – Treatment groups were randomized to receive either 20 mL/kg of Oxyglobin or pRBCT over 4 hours via a central venous catheter. Transfusions were followed by lactated Ringer's solution infusion. Rectal temperature, femoral arterial and mixed venous blood sampling, oscillometric blood pressure, and subjective assessment of patient status (habitus), and appetite were performed at time points 0, 1, 4, 8, 24, 48, and 72 hours.
Main Results – Dogs presented with a hypoalbuminemic alkalosis; hyperchloremic, dilutional acidosis; normotensive tachycardia; pyrexia; depression; and anorexia. Both treatments produced similar results, with the exception of significant differences in pH (4 h); PCO₂ (4 h); hemoglobin (8 h, 24 h); mean arterial pressure (48 h); albumin (4 h, 8 h); habitus (8 h, 48 h); and appetite (24 h). Arterial O₂ content was higher for pRBCT than Oxyglobin at 72 hours, but central venous PO₂ did not differ between groups or over time and was consistently subnormal.
Conclusions – Oxyglobin provides similar overall improvements to pRBCT in dogs with anemia from babesiosis, with respect to blood gas, acid-base and blood pressure, although patients receiving packed cells tended to have more rapid normalization of habitus and appetite.     

Pharmacokinetics of an unfractionated heparin (Liquemin) in the dog after intravenous and subcutaneous application based on heparin activity

In this study pharmacokinetic data for the unfractionated heparin Liquemin were obtained after intravenous and subcutaneous application. Each dosage was examined in 5 healthy, adult Beagle dogs. After intravenous application of 25, 50 and 100 I.U./kg body weight heparin plasma activity of 0.65 +/- 0.15 I.U./ml (mean +/- s), 0.91 +/- 0.10 I.U./ml and 1.94 +/- 0.22 I.U./ml was measured. Subcutaneous applications of 250, 500 or 750 I.U./kg revealed maximum plasma heparin activities of 0.25 +/- 0.10, 0.60 +/- 0.15 and 1.29 +/- 0.24 I.U./ml. The maximum heparin activity in the plasma was observed after 3.8 +/- 1.1 (250 und 500 I.E./kg) or 4.0 +/- 1.0 hours (750 I.E./kg), respectively. Intravenously applicated heparin has a short terminal half-life time (t50) between 22 and 44 minutes. The t50 after subcutaneous application of heparin was distinctly longer. After 250, 500 or 750 I.U./kg the t50 was 3.7 +/- 2.4, 3.5 +/- 1.2 or 5.3 +/- 2.4 hours. Corresponding to this result a lower total clearance (Cltot) was found with increasing doses. Especially the Cltot after subcutaneous injection decreased from 2.08 +/- 0.73 ml/min/kg (250 I.E./kg) to 0.83 +/- 0.27 ml/min/kg (750 I.E./kg). The volume of distribution of heparin corresponded approximately to the plasma volume. The total bioavailability of subcutaneously administered UFH was 53-100% depending on the dosage.     

Comparison of unfractioned and low molecular weight heparin for prophylaxis of coagulopathies in 52 horses with colic: a randomised double-blind clinical trial

REASONS FOR PERFORMING STUDY: Unfractioned heparin (UFH) is widely used for prophylaxis of coagulation disorders, especially in colic-affected horses. However, it is accompanied by certain side effects. OBJECTIVES: To compare the efficacy and side effects of unfractioned and low molecular weight heparin (LMWH) in horses with colic. METHODS: The study was carried out as a randomised, double-blind, controlled clinical trial. Fifty-two horses with colic were treated subcutaneously with either UFH (heparin calcium, 150 iu/kg bwt initially, followed by 125 iu/kg bwt q. 12 h for 3 days and then 100 iu/kg bwt q. 12 h) or LMWH (dalteparin, 50 iu/kg bwt q. 24 h). All horses underwent daily physical examination including assessment of jugular veins, local reaction to heparin injections, haematological evaluation and coagulation profiles over up to 9 days. RESULTS: The type of heparin used did not affect the general behaviour and condition. There were significantly more jugular vein changes in horses treated with UFH. Packed cell volume decreased significantly within the first few days of UFH treatment, but did not change significantly in horses treated with LMWH. Activated partial thromboplastin time (aPTT) and thrombin time (TT) were prolonged in horses treated with UFH but not in those treated with LMWH. CONCLUSIONS: It was concluded that, in comparison to UFH, LMWH has markedly fewer side effects in horses. POTENTIAL RELEVANCE: Therefore, LMWH is recommended for prophylaxis of coagulation disorders in colic patients.     

Accuracy of an Adrenocorticotropic Hormone (ACTH) Immunoluminometric Assay for Differentiating ACTH-Dependent from ACTH-Independent Hyperadrenocorticism in Dogs

Background: Adrenocorticotropic hormone (ACTH) determination has been used for 30 years to distinguish ACTH-dependent hyperadrenocorticism (ADHAC) from ACTH-independent hyperadrenocorticism (AIHAC) in dogs. However, the few studies that have evaluated its diagnostic accuracy, based in the majority of cases on older assays, have been associated with systematic, but highly variable proportions of misclassified or unclassified cases.
Objective: The purpose of the present study is to evaluate the accuracy of a validated ACTH immunoluminometric assay (ILMA) for differentiating between ADHAC and AIHAC.
Animals: One hundred and nine dogs with hyperadrenocorticism were included: 91 with ADHAC and 18 with AIHAC.
Methods: Retrospective study. Dogs displaying feedback inhibition after the dexamethasone suppression test, adrenal symmetry, or both were considered to have ADHAC. AIHAC was demonstrated by adrenal tumor histology. For each group, ACTH determination by ILMA was reviewed.
Results: In the ADHAC group, plasma ACTH measurements ranged between 6 and 1250 pg/mL (median, 30 pg/mL). In the AIHAC group, all ACTH concentrations were below the lower quantification limit of the assay (<5 pg/mL). The 95% confidence interval was 85–100% for sensitivity and 97–100% for specificity in AIHAC diagnosis.
Conclusion and Clinical Importance: No overlap in ACTH concentrations was observed between dogs with ADHAC and dogs with AIHAC. The use of a new technique with high analytical sensitivity made it possible to use a low threshold (5 pg/mL), avoiding the misclassification of some ADHAC cases with low, but quantifiable concentrations of ACTH. The assessment of ACTH concentrations by ILMA is an accurate tool for differentiating between ADHAC and AIHAC.     

Effect of Thyroxine Supplementation on Glomerular Filtration Rate in Hypothyroid Dogs

Background: Glomerular filtration rate (GFR) is decreased in humans with hypothyroidism, but information about kidney function in dogs with hypothyroidism is lacking.
Hypothesis: Hypothyroidism influences GFR in dogs. The objective of this study was to assess GFR in hypothyroid dogs before implementation of thyroxine supplementation and after re-establishing euthyroidism.
Animals: Fourteen hypothyroid dogs without abnormalities on renal ultrasound examination or urinalysis.
Methods: Blood pressure and GFR (measured by exogenous creatinine clearance) were measured before treatment (T0, n = 14) and at 1 month (T1, n = 14) and at 6 months (T6, n = 11) after beginning levothyroxine supplementation therapy (20 μg/kg/d, PO). The response to therapy was monitored at T1 by measuring serum total thyroxine and thyroid stimulating hormone concentrations. If needed, levothyroxine dosage was adjusted and reassessed after 1 month. Statistical analysis was performed using a general linear model. Results are expressed as mean ± standard deviation.
Results: At T0, the average age of dogs in the study group was 6.3 ± 1.4 years. Their average body weight decreased from 35 ± 18 kg at T0 to 27 ± 14 kg at T6 ( P < .05). All dogs remained normotensive throughout the study. GFR increased significantly with levothyroxine supplementation; the corresponding results were 1.6 ± 0.4 mL/min/kg at T0, 2.1 ± 0.4 at T1, and 2.0 ± 0.4 at T6 ( P < .01).
Conclusion: GFR was <2 mL/min/kg in untreated hypothyroid dogs. Re-establishment of a euthyroid state increased GFR significantly.     

Cardiac Troponin I Is Associated with Severity of Myxomatous Mitral Valve Disease, Age, and C-Reactive Protein in Dogs

Background: Concentrations of cardiac troponin I (cTnI) and C-reactive protein (CRP) might be associated with cardiac remodeling in dogs with myxomatous mitral valve disease (MMVD). Age- and sex-dependent variations in cTnI concentration have been described.
Objective: To investigate whether plasma concentrations of cTnI and CRP are associated with severity of MMVD, and investigate potential associations of dog characteristics on cTnI and CRP concentrations.
Animals: Eighty-one client-owned dogs with MMVD of varying severity.
Methods: Dogs were prospectively recruited for the study. Dogs were classified according to severity of MMVD. Plasma cTnI was analyzed by a high sensitivity cTnI assay with a lower limit of detection of 0.001 ng/mL, and plasma CRP was analyzed by a canine-specific CRP ELISA.
Results: Higher cTnI concentrations were detected in dogs with moderate (0.014 [interquartile range 0.008–0.029] ng/mL, P = .0011) and severe (0.043 [0.031–0.087] ng/mL, P < .0001) MMVD, compared with healthy dogs (0.001 [0.001–0.004] ng/mL). Dogs with severe MMVD also had higher cTnI concentrations than dogs with mild (0.003 [0.001–0.024] ng/mL, P < .0001) and moderate ( P = .0019) MMVD. There were significant associations of age, CRP, heart rate, and left ventricular end-diastolic diameter, on cTnI concentration C-reactive protein did not differ among severity groups, but was significantly associated with cTnI, breed, and systolic blood pressure on CRP concentration.
Conclusions and Clinical Importance: Analysis of cTnI concentration has potential to increase knowledge of overall cardiac remodeling in dogs with MMVD. However, effect of age on cTnI needs consideration when assessing cTnI.     

Concentrations of acute-phase proteins in dogs with steroid responsive meningitis-arteritis

Background: Measurement of concentrations of acute-phase proteins (APPs) is used as an aid in the diagnosis of a variety of diseases in animals.
Objective: To determine the concentration of APPs in dogs with steroid responsive meningitis-arteritis (SRMA) and other neurologic diseases.
Animals: One hundred and thirty-three dogs with neurologic diseases, 6 dogs with sepsis, and 8 healthy dogs were included in the study. Thirty-six dogs had SRMA (31 of which had monitoring), 14 dogs had other meningoencephalitides (ME), 32 had disk disease (IVDD/DLSS), 26 had tumors affecting the central nervous system (TCNS), and 25 had idiopathic epilepsy (IE).
Methods: Prospective, observational study: C-reactive protein (CRP), α₂-macroglobulin (AMG), and albumin concentrations were determined in the serum or plasma. CRP was also measured in the cerebrospinal fluid.
Results: Serum CRP was significantly higher in dogs with SRMA (     = 142 μg/mL ± 75) and sepsis (     = 114 μg/mL ± 67) in comparison with dogs with other neurologic diseases (     = 2.3–21 μg/mL; P < .001). There was no significant difference detected in AMG between groups. Serum albumin concentration was significantly lower ( P < .01) in dogs with SRMA (     = 3.2 g/dL ± 0.41) than in other groups (     = 3.6–3.9 g/dL). Serum CRP concentration of SRMA dogs correlated with alkaline phosphatase levels ( r = 0.515, P = .003).
Conclusions and Clinical Importance: CRP concentrations in serum are useful in diagnosis of dogs with SRMA. Serum CRP could be used as a monitoring parameter in treatment management of these dogs.