Influencing factors of ADP-induced, epinephrine-induced and ristomycin-induced platelet aggregation in dogs.

Functional defects of platelets are often studied by in-vitro aggregation tests with chemical compounds such as ADP, epinephrine and ristomycin (ristocetin). The aim of the present work was to investigate the effect of some diseases and that of nonsteroidal anti-inflammatory drug treatment on platelet aggregation in dogs. The examination had been carried out on 115 dogs by a Carat TX4 optical aggregometer (Entec GmbH, Ilmenau, Germany) first used in veterinary practice. The dogs were divided in three groups: healthy (control) dogs (n = 43), diseased dogs with normal haemostasis profile (n = 44), and dogs suffering from arthropathies with normal haemostasis profile treated with the nonsteroidal anti-inflammatory drugs ketoprofen or carprofen (n = 21). Following establishment of normal platelet aggregation curves in healthy dogs we found that some diseases such as diabetes mellitus, Cushing's disease, mastocytoma and lymphoma increased or decreased the aggregation maximum of platelets or caused changes in the feature of the aggregation curve. Carprofen treatment had no effect on platelet aggregation while ketoprofen decreased the aggregation maximum. These results showed that Carat TX4 aggregometer proved a useful instrument in studying platelet aggregation in platelet-rich plasma of dogs. For clinical pathologists it is important to know that the effects of some diseases and nonsteroidal anti-inflammatory drug treatments have to be taken into account when in-vitro platelet aggregation is evaluated. Based on our results and on those of other studies, we think standardization in aggregation methodology is highly recommended in veterinary laboratories.

Evaluation of the effect of ketoprofen and carprofen on platelet function in dogs studied by PFA-100 point-of-care analyser.

The effect of two nonsteroidal anti-inflammatory drugs (carprofen and ketoprofen) on platelet adhesion and aggregation functions was evaluated by the PFA-100 analyser (Dade-Behring, CA, U.S.A.) using its collagen-adenosine diphosphate (ADP) and collagen-epinephrine (EPI) cartridges. The function of platelets was evaluated in 55 healthy dogs, in 7 dogs treated with ketoprofen and in 31 dogs treated with carprofen in a therapeutic dose for minimum 5 days. The therapeutic doses of carprofen had no effect on the closure time of PFA-100 (which is the marker of platelet function) but ketoprofen caused a significant increase when using collagen-EPI stimulation The closure times for both the healthy (control) and the treated dogs using EPI cartridges were often longer than the upper default cut-off point (300 sec) of the device. The PFA-100 analyser with collagen-ADP cartridges could be a useful tool for veterinary applications including the evaluation of platelet aggregation in dogs treated with NSAIDs. The upper cut-off point of PFA-100 might be extended.

Morphological evaluation of canine platelets on Giemsa- and PAS-stained blood smears.

The morphology of canine platelets (changes in size, shape, staining characteristics, degree of activation and clump formation, distribution of granules, appearance of vacuoles on Giemsa-stained smears) was investigated in 20 healthy control and 181 diseased dogs. In the group of the sick dogs 84 animals suffered from disorders affecting directly the haematological parameters or the haematopoietic organs such as bleeding, thymic haemorrhage, haemolytic disorders, lymphoma, immune-mediated thrombocytopenia, and other 97 dogs were affected by other diseases (hepatopathy, nephropathy, hepatic, splenic or intestinal neoplasm, skin diseases, diabetes mellitus, Cushing's syndrome, sepsis). The alterations found in platelet morphology were not specific for any disorder. The most common platelet abnormalities were polychromasia and the presence of giant platelets. These changes occurred in a high number in disorders accompanied by bleeding or haemolysis. Anisocytosis was the most frequent finding in hepatic, splenic or intestinal neoplasms and in certain endocrinopathies. Microcytosis was observed in immune-mediated thrombocytopenia, hepatic neoplasms and endocrine disorders. Extreme platelet activation was common in haemolysis, hepatopathies, neoplastic diseases and sepsis. Vacuolisation was present in thymic haemorrhage, pancreatitis, diabetes mellitus and Cushing's syndrome. A new morphologic phenomenon, i.e. a ring-like formation of granules, was described in the cytoplasm of the platelets both in healthy and diseased animals. In addition, two forms of pathologic granulation were also described for the first time in Giemsa-stained blood smears: the pseudonuclear and the spot-like formation of granules, which were observed especially in disorders affecting the blood cells. The granulation and morphological characteristics of platelets on smears stained by periodic acid-Schiff reaction (PAS) were also studied. Three localisations of granulation were observed, such as peripheral, eccentric and diffuse. The ratio of PAS-positive and -negative platelets was evaluated in several diseases. Our findings support the diagnostic value of platelet evaluation by light microscopy and help clinicians/clinical pathologists to understand why morphologic changes of thrombocytes might be expected in several diseases.