Effects of dipotassium EDTA and lithium heparin on hematological indexes CAS NO.25102-12-9

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Blood analysis is a very important and powerful diagnostic tool in animal health and welfare control. It is usually carried out in higher vertebrates, and its reference value has been established, but fish hematology still needs further research. Many internal and environmental factors have a far-reaching impact on the hematological value of fish, making it difficult to determine the reference value. In addition, due to the short coagulation time, fish blood usually needs to add anticoagulants. The selection of anticoagulants is very important to obtain reliable blood test values. In this study, the effects of two common anticoagulants, K ＾ EDTA (1.8 mg / ml) and heparin lithium (18 IU / ml), on the hematological values of cultured spotted trout brown trout were investigated.

The results of basic hematological analyses, such as compacted cell volume (PCV), hemoglobin concentration (Hgb), red blood cell count (RBC), mean red blood cell hemoglobin (MCH), mean red blood cell volume (MCV), mean red blood cell hemoglobin concentration (MCHC), and white blood cell count (WBC), were compared between K 2 EDTA and heparin lithium. Statistically significant differences were observed in PCV, MCV and MCHC, while HGB, RBC, MCH and WBC did not show this difference. These results suggest that heparin lithium provides more reliable results because the red blood cells in the samples treated with K ＾ 2 ＾ EDTA tend to swell.

The salts of ethylenediamine tetraacetic acid (EDTA) are recommended by the International Committee for hematological Standardization (ICSH) and the Institute of clinical and laboratory standards (CLSI) as the preferred anticoagulant for blood cell counting. EDTA acts as a chelating agent for Ca ＾ 2 + and Mg ＾ 2 + ions. It combines calcium and is essential for the enzymatic reaction in the coagulation cascade to produce anticoagulant effect with minimal effect on cell morphology. EDTA can most reliably preserve the cells on the stained blood membrane. It is often used in routine hematological tests of humans, mammals and other vertebrates. It has also achieved some success in fish. However, EDTA salt has some disadvantages. In some cases, they can cause blood hemolysis, especially in specimens anesthetized with unbuffered tetracaine methanesulfonate (MS 222).

Heparin salt is mainly used in clinical biochemistry. It prevents the conversion of prothrombin to thrombin (factor II), thereby inhibiting the conversion of fibrinogen to fibrin. According to the few reports of hemolysis in EDTA preserved blood samples of some reptiles and birds, it is recommended to use heparin in the blood cell count of animals with red blood cells. For this reason, many authors use heparin as the preferred anticoagulant in fish hematology. However, heparin does not completely prevent leukocyte and platelet aggregation, which leads to sample coagulation and blue background staining of Wright stained blood smears. In addition, it is more expensive than other anticoagulants.

Some researchers have observed significant differences in HGB and RBC obtained from EDTA and heparin salt. Heparin lithium is more suitable for hematological evaluation of spawning of cultured trout than ethylenediamine tetraacetate. Heparin does not affect the size of red blood cells, thus minimizing the possibility of hemolysis. Compared with heparin, blood smears made from blood treated with EDTA are of higher quality. Based on the current research, we suggest that k2edta can be carefully used for the determination of blood cell count and HGB. In order to better understand the effect of anticoagulants on hematological parameters, further research is needed.