Veterinary and Experimental Toxicology, July 2017 (1-4) Electrometric determination of erythrocyte and plasma cholinesterase activities in animals
F. K. Mohammad Ministry of Higher Education and Scientific Research, Baghdad, Iraq e-mail: fkmohammad@vetmedmosul.org
Chemicals: 7.5% aqueous solution of acetylthiocholine iodide, sodium barbital, potassium dihydrogen phosphate, sodium chloride, double distilled water. Barbital-Phosphate buffer: 1.237 g sodium barbital, 0.163 g potassium dihydrogen phosphate and 35.07 g sodium chloride/ L distilled water, pH 8.1. Equipment: Water bath, pH meter
1. Collect venous blood samples in heparinized test tubes. 2. Separate the plasma from erythrocytes by centrifugation. 3. To a 10-ml beaker add 3 ml distilled water, 0.2 ml plasma, erythrocytes or even whole blood. 4. Add 3 ml of barbital-phosphate buffer solution. 5. Measure the pH of the mixture (pH1). 6. Add 0.10 ml of 7.5% acetylthiocholine iodide to the mixture. 7. Incubated at 37 ยบC for 30 minutes. 8. Measure the pH of the mixture (pH2).
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Veterinary and Experimental Toxicology, July 2017 (1-4) 9. Calculated cholinesterase activity: Cholinesterase activity (∆pH/30 min) = (pH1 – pH2) - ∆ pH of blank Notes: The blank is without sample. The unit of activity is ∆ pH/30 min.
References and further reading 1. Mohammad FK, Al-Baggou BK, Naser AS and Fadel MA. In vitro inhibition of plasma and brain cholinesterases of growing chicks by chlorpyrifos and dichlorvos. Journal of Applied Animal Research 2014. 2. Mohammad FK, Mousa YJ, Al-Zubaidy MHI, Alias AS. Assessment of diphenhydramine effects against acute poisoning induced by the organophosphate insecticide dichlorvos in chicks. Human and Veterinary Medicine Bioflux 2012; 4: 6-13. 3. Al-Zubaidy MHI, Mousa YJ, Hasan MM, Mohammad FK. Acute toxicity of veterinary and agricultural formulations of organophosphates dichlorvos and diazinon in chicks. Archives of Industrial Hygiene and Toxicology 2011; 62: 317322. 4. Alias AS, Al-Zubaidy MHI, Mousa YJ, Mohammad FK. Plasma and whole brain cholinesterase activities in three wild bird species in Mosul, Iraq: in vitro inhibition by insecticides. Interdisciplinary Toxicology 2011; 4: 144-148. 5. Al-Baggou BK, Naser AS, Mohammad FK. Hydrogen peroxide potentiates organophosphate toxicosis in chicks. Human and Veterinary Medicine Bioflux 2011; 3(2):142-149. 6. Mohammad FK. Laboratory Guide in Toxicology. 2nd edition. Printed by Mizzou Media, University of Missouri, MO, USA., 2010. 7. Mohammad FK, Al-Badrany YM, Al-Jobory MM. Acute toxicity and cholinesterase inhibition in chicks dosed orally with organophosphate insecticides. Archives of Industrial Hygiene and Toxicology 2008; 59: 145-151. 8. Al-Badrany YMA Mohammad FK. Effects of acute and repeated oral exposure to the organophosphate insecticide chlorpyrifos on open-field activity in chicks. Toxicology Letters 2007; 174: 110-116.
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Veterinary and Experimental Toxicology, July 2017 (1-4)
9. Mohammad FK. Review of a practical electrometric method for determination of blood and tissue cholinesterase activities in animals. Vet Scan 2007; 2:1-12. 10. Mohammad FK, Alias AS, Faris GA-M, Al-Baggou BKh. Application of an electrometric method for measurement of blood cholinesterase activities in sheep, goats and cattle treated with organophosphate insecticides. Journal of Veterinary Medicine (A) 2007; 54: 140-143. 11. Mohammad FK, Al-Zubaidy MHI, Alias AS. Electrometric determination of erythrocyte, plasma and whole blood cholinesterase activities in sheep, goats and cattle and their in vitro inhibition by anticholinesterase insecticides. Journal of Pharmacology and Toxicology 2007; 2:131-141. 12. Mohammad FK, Al-Baggou' BK, Alias AS, Faris GA-M. Application of an electrometric method for measurement of in vitro inhibition of blood cholinesterases from sheep, goats and cattle by dichlorvos and carbaryl. Veterinarni Medicina 2006; 51: 45-50. 13. Alias AS, Mohammad FK. Electrometric measurement of plasma and tissue cholinesterase activities of four wild birds in Iraq. Journal of Biological Research 2005; 4: 197-202. 14. Mohammad FK, Faris GA-M, Alias AS, Al-Baggou' BKh. Blood cholinesterase activities in cattle, sheep and goats measured by a modified electrometric method. Journal of Animal and Veterinary Advances 2005; 4: 923-926. 15. Mohammad FK, Al-Baggou' B. Electrometric cholinesterase determination in poultry treated with dichlorvos and carbaryl. Online Journal of Veterinary Research 2005; 9: 1-5. 16. Mohammad FK, Faris GA-M, Shindala MK. Comparative antidotal effects of diphenhydramine and atropine against dichlorvos-induced acute toxicosis in rats. Veterinarski Arhiv 2002; 72: 19-28. 17. Al-Baggou' BKh, Mohammad FK. Antagonism of methomyl-induced toxicosis by diphenhydramine in rats. Environmental Toxicology and Pharmacology 1999; 7:119-125. 18. Mohammad FK, Faris GA-M, Al-Kassim NA. A modified electrometric method for measurement of erythrocyte acetylcholinesterase activity in sheep. Veterinary and Human Toxicology 1997; 39: 337-339.
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Veterinary and Experimental Toxicology, July 2017 (1-4)
19. Faris GA-M, Mohammad FK. Prevention and treatment of dichlorvos-induced toxicosis in mice by diphenhydramine. Veterinary and Human Toxicology 1997; 39: 22-25. 20. Mohammad FK, St Omer VEV. Modifications of Michel's electrometric method for rapid measurement of blood cholinesterase activity in animals: a minireview. Veterinary and Human Toxicology 1982; 24: 119-121.
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