Effects of sampling conditions on selected blood variables of rainbow trout, Salmo gairdneri Richardson

Abstract

The effects of two methods of specimen immobilization (MS 222 anaesthesia and stunning), two types of anticoagulant (EDTA and heparin), two storage temperature ranges (0-2" C and 22.-25" C) and four sample storage periods (0, 1, 3, and 24 h) on the haemoglobin, haematocrit, plasma and packed cell sodium, potassium and chloride ion concentrations and packed cell ATP levels of rainbow trout were examined. Stored samples exhibited increases in cell volume, net transfer of sodium and chloride from plasma into cells, net loss ofpotassium to plasma and rapid depletion of ATP. Room temperature conditions and prolonged storage exacerbated these changes. Use of EDTA, particularly in combination with MS 222, frequently led to haemolysis. Least change in most variables was observed in samples drawn from stunned specimens, treated with heparin and refrigerated before use or preparation for deep cold storage.

Na2EDTA anticoagulant impaired blood samples from the teleost Piaractus mesopotamicus

“…Walencik & Witeska (2007) observed higher fragility in common carp erythrocytes to hipotonic solution since the lesser concentration of Na 2 EDTA tests (0.01mg ml -1 ) and a positive relation of the osmotic fragility and the increase of the anticoagulant concentrations. Similar results were described in Blennius pholis (Mainwaring & Rowley 1985), Oncorhynchus mykiss (Korcock et al 1988), Oreochromis niloticus (Ekanem et al 2012), Pseudoplatystoma reticulatum x P. corruscans (Ishikawa et al 2010) and Colossoma macropomum (Pádua et al 2012).…”

“…The sampling of the blood of each fish was realized through veno-caudal puncture, using sterile syringes (3mL) and hypodermic needles (25x7mm), containing no anticoagulants, as recommended by Ishikawa et al (2010). Trying to not promote any hemolitic effects besides the possibly caused by the anticoagulants, we decided to use no anesthetic chemicals (Korcock et al 1988). The blood collected, circa of 0.5mL, was quickly placed in four polyethylene tubes (1.5mL): The first and second contained 15μL of NA 2 EDTA, at a concentration of 3% (0.3mg mL -1 of blood) and 10% (1mg mL -1 of blood), respectively.…”

Heparina e Na2EDTA como anticoagulantes para surubim híbrido (Pseudoplatystoma reticulatum x P. corruscans): eficácia e alterações hematológicas

“…Após a captura, os peixes foram contidos mecanicamente com pano umedecido e realizada venopunção dentro de poucos segundos, de forma a minimizar os efeitos do estresse agudo provocado pela captura, visto que a contenção química por fármacos anestésicos pode determinar a ocorrência de hemólise em peixes (KORCOCK et al, 1988). Procedeu-se à venopunção caudal, com auxílio de seringas descartáveis de 5mL e agulhas hipodérmicas 30x8mm desprovidas de anticoagulante, sendo puncionados 2,5mL por indivíduo.…”

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“…Erythrocyte swelling may be due to an increase in pCO 2 and acidification due to treatment with acidic EDTA salt (Smit et al, 1977). Erythrocytes swelling in EDTA treated samples was also observed by Blaxhall (1973) and Korcock et al (1988) in fish, and by Olsen et al (2001) in mammal blood. In contrast, a decrease in hematocrit values in mammal blood samples with EDTA compared to heparinized blood was reported (Morris et al, 2002), while EDTA did not affect Hct values in green iguana blood (Hanley et al, 2004).…”

“…Moreover, as fish blood tends to clot rapidly and clotting becomes faster when it is warm or is under stress condition related to experimental procedures (Korcock et al, 1988), the use of anticoagulants is necessary to obtain reliable results of blood analyses. The diagnostic techniques applied in fish hematology are usually adapted methods developed in human hematology.…”