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March 1997

Diagnosis of Bleeding Disorders

Determining the cause in a patient presented for excessive bleeding can be a diagnostic challenge. In addition to a thorough physical examination, a recent and prior history or any relevant family history should be obtained. If the animal has been neutered or undergone some other type of surgery without evidence of abnormal bleeding, this points towards a recent acquired cause of hemorrhage rather than a congenital or hereditary problem. Breed experience [e.g. Doberman pinschers with their high prevalence of von Willebrand’s disease (vWD)] is also important. Use of drugs that impair hemostasis, potential toxin exposure such as to rodenticides, or vaccinations within the previous 30 days may be inciting causes. Pre- existing liver or kidney disease could play a role as well. Asymptomatic carriers of bleeding diseases such as vWD can express the trait if some other precipitating stress event or exposure occurs. Animals of mostly white coat color or dilutional pigmentation may be at increased risk to bleed excessively when hemostatically challenged because of their relative platelet dysfunction.

The character of bleeding presents valuable diagnostic clues. Petechial or ecchymotic hemorrhages are typical of thrombocytopenia; mucosal surface bleeding and excessive hemorrhage or oozing from surgical sites is seen in platelet defects (quantitative and/or qualitative) and vWD; whereas, large surface hematomas and protracted bleeding characterizes congenital disorders like the hemophilias and severe acquired problems such as rodenticide toxicosis and disseminated intravascular coagulation (DIC).

Diagnostic evaluation begins with a basic coagulation profile (platelet count, PT and APTT, fibrinogen level and test for FDP) to which a vWD assay can be added as needed. The bleeding time may also be a valuable tool and can be measured by the mucosal (buccal) method with a template device or by transecting a toe nail (see below). The buccal bleeding time

primarily measures platelet function and can therefore be normal in patients with unstable primary hemostatic plugs or defects in fibrin formation. The toe nail method offers a more complete assessment of hemostasis, although it can be difficult to standardize.

When significant thrombocytopenia is present (platelet counts < 50,000/pl), the laboratory can assess platelet size and volume. Predominantly small platelets (MPV < 5.0 fl) are characteristic of immune-mediated platelet destruction, whereas mostly large platelets (MPV > 9.2 fl) signify active thrombopoiesis and tend to be more adhesive. Please indicate on your Test Request Form or by phone request that you wish to have platelet size evaluated, if the platelet count is very low.

Toe Nail (Transection) Bleeding Timea clinical assessment of bleeding potential in dogs and cats.

With the animal in lateral recumbency, either awake or anesthetized, cut one or more toe nails too short with a sharp guillotine-type toe nail clipper. Make a clean transection of the nail just into the quick (becomes reproducible with practice). Let the nail bleed freely undisturbed and time until bleeding stops. Normal bleeding times are up to 5 minutes in dogs and 2 1/2-3 minutes in cats. A prolonged bleeding time or rebleeding once it has stopped is considered abnormal. Bleeding times measured by this technique are sensitive to defects in vascular contraction, platelet function and coagulation. This test can be useful for:

  • pre-surgical assessment of bleeding potential;
  • evaluation of response to therapy in bleeding animals;
  • determination of degree of bleeding risk (e.g. slight prolongation of 1-3 minutes may be manageable conservatively without transfusion; moderate to severe prolongation of 3 or more minutes usually requires treatment before surgery or to alleviate clinical signs; bleeding time endpoints over 10 minutes are significantly prolonged and require transfusional therapy).

Lip (Mucosal) Bleeding Time – a clinical assessment of bleeding potential in horses and farm animals.

With the animal standing quietly, make a cut about 1.5 cm long on the inside surface of the lower lip. Use a fresh scalpel blade wrapped with adhesive tape to expose about 3 mm of the blade tip. Hold the blade between thumb and forefinger at the level of the tape to control the depth of incision. Incise with steady pressure. Let bleed freely and time until bleeding stops. Normal values are up to 8 or 9 minutes. Prolonged bleeding or rebleeding once it has stopped is abnormal. Bleeding times measured with this template-type technique are primarily sensitive to defects of platelet function (quantitative and/or qualitative).

The basic coagulation profile evaluates intrinsic (APTT) and extrinsic (PT) coagulation activity and the presence of DIC (fibrinogen and FDP tests). It will not diagnose cases of vWD. Coagulation laboratories have historically used the thrombin time test to screen for the presence of fibrinogen-fibrin degradation products. However, the thrombin reagents used today are insensitive to the presence of FDP based on a thorough evaluation recently performed at Antech. These reagents are useful, however, for determining fibrinogen concentration based on the thrombin clottability technique. Consequently, Antech will no longer offer the thrombin time test but will replace it with a fibrinogen activity assay and FDP measurements (D-dimer test).

Special coagulation factor assays can be performed on a send-out basis, if the basic coagulation profile results and clinical or family history so warrant. Please contact the laboratory for specific instructions to prepare and submit these samples, as they need to be sent frozen with dry ice. The most common problems encountered with samples submitted for coagulation evaluation are as follows:

  • Clotted or hemolyzed specimens. Clots of any size and moderate to significant hemolysis will invalidate coagulation and vWD tests.
  • Wrong anticoagulant – EDTA (LTT) or heparin (GTT) instead of citrate (BTT).
  • No anticoagulant – clotted serum tube or FDP tube used instead of BTT.
  • Improperly filled Vacutainer tube. Sample tubes filled to only half or less are overdiluted with citrate and will likely produce invalid (prolonged) results.
  • Incomplete or missing patient descriptors (e.g., species, age, sex, breed, neutered or intact).
  • Specimens held too long prior to pick up or mailed- in with more than a 24-hour receiving delay can produce erroneous coagulation results. In such cases we prefer that BTT samples be spun down soon after collection, and the plasma removed and placed into an empty plastic or silicone-coated glass tube prior to submission. A LTT sample is still required for the CBC/platelet count portion of this profile.

The most important points in obtaining valid specimens for coagulation assays are a clean venipuncture with rapid blood flow into the collection syringe or Vacutainer tube, and the correct volume of blood to fill the tube.

A recent clinical case illustrates the value of coagulation testing. A one-year-old spayed female cat was admitted to an emergency clinic for evaluation of severe anemia (PCV 7%). She was FIA positive for hemobartonellosis. A routine coagulation profile revealed: PT 14.0 sec (normal range 9-12 sec).

APTT > 60 sec (18-22 sec)
Thrombin time 5.5 sec (5-9 sec)
Platelet count 33,000/pl (200-500,000/pl)
Fibrinogen < 100 mg/dl (100-400 mg/dl)

Three weeks later after treatment for the hemobartonella, followup testing showed PCV had risen to 17%, 16 nucleated RBC/100 WBC, FIA was negative but the coagulation profile remained abnormal:

PT 10.1 sec. APTT > 60 sec.
Thrombin time 6.6 sec.
Platelet count 130,000/pl
Fibrinogen 319 mg/dl

Because there was no overt bleeding tendency observed in this patient and it was a cat with persistently long APTT, the presumptive diagnosis of the coagulopathy was Hageman trait (factor XII deficiency). This condition is relatively common in cats, and is usually an incidental finding not associated with excessive bleeding unless some other hemostatic defect coexists (e.g., thrombocytopenia). Citrated plasma was submitted in dry ice to the Comparative Coagulation Section at Cornell University’s Diagnostic Laboratory and revealed a factor XII level of 1.5% (normal range 60-150%). This confirmed the diagnosis as Hageman trait.

 
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