Teg5000 thromboelastograph hemostasis analyzer

TEG assays were adapted from Zeng et al. [32 (link)]. Briefly, TEG assays were performed on a TEG^®5000 Thromboelastograph Hemostasis Analyzer at 37 °C with TEG Analytical Software^® Version 4 (Haemonetics Corporation, Braintree, MA, USA) run in triplicate with one additional no-microplastic control cup per run. As noted, volumes of thrombin, fibrinogen, PBS, and microplastics were added to clear TEG cups (360 µL total). Sample preparation for TEG was carried out in the same manner as turbidity, keeping concentrations consistent across assays. TEG assays were allowed to run to completion. R-time, angle, maximum amplitude (TEG^max), and time to maximum amplitude (TEG^time) were recorded from the software.

A complete blood count was performed with EDTA-anticoagulated whole blood using the Sysmex XT2000 (Sysmex, Kobe, Japan). Samples were thoroughly inverted and then analyzed as recommended by the manufacturer.
Thromboelastography (TEG) was performed with recalcified citrate whole blood using the TEG 5000 Thromboelastograph Hemostasis Analyzer (Haemonetics GmbH, Boston, MA, USA). The cup was placed according to the manufacturer’s instructions and 20 µL of 0.2 M CaCl2 was pipetted in the cup. One milliliter of the citrate whole blood was transferred to a kaolin tube (Haemonetics GmbH) and mixed by gentle inversion 5 times. Immediately afterwards, 340 µL of the kaolin citrate whole blood mixture was pipetted to the CaCl2 solution in the cup and the thromboelastograph was started.
The coagulation parameters, prothrombin time (PT), activated partial thrombin time (aPTT) and thrombin time, were analyzed using the STA Compact Max3 (Asnières-sur-Seine, France) according to the manufacturer’s instructions.
Dog breeders/owners were contacted via phone and asked for conspicuities regarding bleeding tendencies and vision.