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Licensed Unlicensed Requires Authentication Published by De Gruyter August 5, 2014

Influence of extracellular media’s ionic strength on the osmotic stability of Sahel goat erythrocytes

  • Nanacha Afifi Igbokwe and Ikechukwu Onyebuchi Igbokwe EMAIL logo

Abstract

Background: Heparinised blood was exposed to osmotic lysis in hypotonic buffered saline to evaluate erythrocyte membrane stability. When K3 EDTA blood was used, it added more to the ionic content of blood than heparin. The influence of suspending media’s ionic strength on the osmotic stability of Sahel goat erythrocytes was investigated by replacing the ionic saline with non-ionic saccharide (sucrose or glucose) and assessing the effect of using EDTA blood instead of heparinised blood.

Results: The erythrocyte osmotic fragility curve in saline was hyperbolic even when the ionic concentration was reduced by 50% with saccharides. Haemolysis was higher with EDTA than heparinised blood at saline concentrations of 90 and 150–180 mosmol/L. The fragility curve was sigmoidal and shifted to the left when saline was completely substituted with a saccharide. The non-ionic saccharides increased erythrocyte osmotic resistance linearly (r=0.88; p<0.02) from median to minimal hyposmolarities (150–300 mosmol/L) and reduced the osmolyte concentration at median fragility by 36%. No effect occurred at <30–120 mosmol/L and >90% fragility; and saccharide concentrations were almost non-lytic at comparable saline concentrations evoking <10% haemolysis. Fragilities were neither affected by period (30–60 min) of incubation nor the type of saccharide used.

Conclusions: In this study, the variation in osmotic stability of caprine erythrocytes was linked to ionic strength of the suspending extracellular media which seemed to exert an influence through transmembrane ion fluxes and regulatory volume changes in erythrocytes.


Corresponding author: Ikechukwu Onyebuchi Igbokwe, Strategic Animal Research Group, Department of Veterinary Pathology, University of Maiduguri, PO Box 8000, Maiduguri, Nigeria, Phone: +234 834811622, E-mail:

Conflict of interest statement

Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

References

1. Parpart AK, Lorenz PB, Parpart ER, Gregg JR, Chase AM. The osmotic resistance (fragility) of human red cells. J Clin Invest 1947;26:636–40.10.1172/JCI101847Search in Google Scholar

2. Inaba M, Messick JB. Erythrocyte membrane defects. In: Weiss DJ, Wardrop KJ, editors. Schalm’s veterinary hematology, 6th ed. Ames: Blackwell Publishing, 2010:187–95.Search in Google Scholar

3. Wright IG. Osmotic fragilty of erythrocytes in acute Babesis argentina and Babesia bigemina infections in splectomized Bos Taurus calves. Res Vet Sci 1973;15:299–305.10.1016/S0034-5288(18)33799-8Search in Google Scholar

4. Oyewale JO. Studies on the erythrocyte osmotic fragility of rats infected with Trypanosoma brucei. Anim Technol 1987;38:219–28.Search in Google Scholar

5. Mijares A, Vivas J, Abad C, Betancourt M, Piñero S, Proverbio F, et al. Trypanosoma evansi: Effect of experimental infection on the osmotic fragility, lipid peroxidation and calcium-ATPase activity of rat red blood cells. Exp Parasitol 2010;124:301–5.10.1016/j.exppara.2009.11.002Search in Google Scholar

6. Kobo PI, Ayo JO, Aluwong T, Zezi AU, Maikai V, Ambali SF. Flavonoid mixture ameliorates increase in erythrocyte osmotic fragility and malondialdehyde concentration induced by Trypanosoma brucei brucei-infection in Wistar rats. Res Vet Sci 2014;96:139–42.10.1016/j.rvsc.2013.10.005Search in Google Scholar

7. Silva IM, Hubsch C, Ysern-Caldentey M. Erythrocyte osmotic fragility and cation concentrations during experimentally induced bovine anaplasmosis. Comp Biochem Physiol Part A: Physiol 1989;94:455–9.10.1016/0300-9629(89)90120-5Search in Google Scholar

8. Jain NC. Essentials of veterinary hematology. Philadelphia: Lea and Febiger, 1989:203–6.Search in Google Scholar

9. Lessler MA, Walters MI. Erythrocyte osmotic fragility in the presence of lead or mercury. Exp Biol Med 1973;142:548–53.10.3181/00379727-142-37064Search in Google Scholar

10. Seeman P, Weinstein J. Erythrocyte membrane stabilization by tranquilizers and antihistamines. Biochem Pharmacol 1966;15:1737–52.10.1016/0006-2952(66)90081-5Search in Google Scholar

11. Seeman P, Kwant WO, Sauks T, Argent W. Membrane expansion of intact erythrocytes by anesthtics. Biochim Biophys Acta 1969;183:490–8.10.1016/0005-2736(69)90163-1Search in Google Scholar

12. Bilto YY. Rheological action of aspirin on human erythrocytes. Clin Hemorrheol Microcirc 1990;20:159–65.Search in Google Scholar

13. Ambali SF, Ayo JO, Ojo SA, Esievo KA. Ameliorative effect of vitamin C on chlorpyrifos-induced increased erythrocyte fragility in Wistar rats. Hum Exp Toxicol 2010;30:19–24.10.1177/0960327110368415Search in Google Scholar PubMed

14. Perk K, Frei YF, Herz A. Osmotic fragility of red blood cells of young and mature domestic and laboratory animals. Am J Vet Res 1964;25:1241–8.Search in Google Scholar

15. Bogin E, Massry SG, Levi J, Djaldeti M, Bristol G, Smith J. Effect of parathyroid hormone on osmotic fragility of human erythrocytes. J Cin Invest 1982;69:1017–25.10.1172/JCI110505Search in Google Scholar PubMed PubMed Central

16. Oyewale JO, Durotoye LA. Osmotic fragility of erythrocytes of two breeds of domestic fowl in the warm humid tropics. Lab Anim 1998;22:250–4.Search in Google Scholar

17. Oladele SB, Ayo JO, Ogundipe SO, Esievo KA. Seasonal and species variations in erythrocytes osmotic fragility of indigenous poultry species in Zaria, Northern Guinea Savannah zone of Nigeria. Poult Sci 2003;56:1842–6.Search in Google Scholar

18. Penha-Silva N, Firmino CB, Reis FG, Huss JC, Souza TM, Freitas MV, et al (2007) Influence of age on the stability of human erythrocyte membranes. Mech Ageing Dev 2007;128:444–9.10.1016/j.mad.2007.06.007Search in Google Scholar

19. Yucel R, Ozdemir S, Dariyel N, Toplan S, Akyolku MC, Yigit G. Erythrocyte osmotic fragility and lipid peroxidation in experimental hyperthyroidism. Endocrine 2009;36:498–502.10.1007/s12020-009-9251-6Search in Google Scholar

20. Oyewale JO. Osmotic fragility of erythrocytes of West African Dwarf sheep and goats: effects of temperature and pH. Br Vet J 1991;147:163–70.10.1016/0007-1935(91)90107-XSearch in Google Scholar

21. Oyewale JO. Effect of storage of blood on the osmotic fragility of mammalian erythrocytes. J Vet Med A 1993;40:258–64.10.1111/j.1439-0442.1993.tb00626.xSearch in Google Scholar

22. Oyewale JO, Okewumi TO, Olayemi FO. Haematological changes in West African Dwarf goats following haemorrhage. J Vet Med A 1997;44:619–24.10.1111/j.1439-0442.1997.tb01148.xSearch in Google Scholar

23. Kafka M, Yermiahu T. The effect of EDTA as an anticoagulant on the osmotic fragility of erythrocytes. Clin Lab Haematol 1998;20:213–6.10.1046/j.1365-2257.1998.00014.xSearch in Google Scholar

24. Mafuvadze B, Erlwanger KH. The effect of EDTA, heparin and storage on the erythrocyte osmotic fragility, plasma osmolality and haematocrit of adult ostriches (Struthio camelus). Vet arhiv 2007;77:427–34.Search in Google Scholar

25. Fourie FR. Effects of anticoagulants on the hematocrit, osmolarity and pH of avian blood. Bull Anim Hlth Prod Afr 1997;51:204–14.Search in Google Scholar

26. Mafuvadze B, Nyanungo M, Saina H, Gorejena B, Mashayamombe T, Erlwanger KH. Deprivation of drinking water for up to 48 h does not affect the osmotic fragility of erythrocytes from captive helmeted guinea fowl (Numida meleagris). Int J Poult Sci 2008;7:59–63.10.3923/ijps.2008.59.63Search in Google Scholar

27. Viscor G, Palomeque J. Method of determing the osmotic fragility curves of erythrocytes in birds. Lab Anim 1982;16:48–50.10.1258/002367782780908797Search in Google Scholar

28. Jandl JH. Leaky red cells Blood 1965;26:367–82.10.1182/blood.V26.3.367.367Search in Google Scholar

29. Woodward CB, Zwaal RF. The lytic behaviour of pure phospholipases A2 and C towards osmotically swollen erythrocytes and resealed ghosts. Bichim Biophys Acta 1972;274:272–8.10.1016/0005-2736(72)90302-1Search in Google Scholar

30. Bowdler AJ, Chan TK. The time course of red cell lysis in hyptonic electrolyte solutions. J Physiol 1969;201:437–52.10.1113/jphysiol.1969.sp008765Search in Google Scholar

31. Winter WP. Mechanism of saponin-induced red cell hemolysis: Evidence for the involvement of aquaporin CHIP28. Blood 1994;84:(Suppl 1–10):[abstract 445].Search in Google Scholar

32. Roelofsen B, Zwaal RF, Comfurius P, Woodward CB, Van Deenen LL. Action of pure phospholipase A2 and phospholipase C on human erythrocytes and ghosts. Biochim Biophys Acta 1971;241:925–9.10.1016/0005-2736(71)90024-1Search in Google Scholar

33. Greene NM. Glucose permeability of human erythrocytes and effects of inhalation anesthetics, oxygen and carbon dioxide. Yale J Biol Med 1965;37:319–30.Search in Google Scholar

34. Guarner V, Alvarez-Buylla R. Erythrocyte and glucose homeostasis in rats. Diabetes 1989;38:410–5.10.2337/diab.38.4.410Search in Google Scholar PubMed

35. Travis SF, Morrison AD, Clements RS, Winegrad AI, Oski FA. Metabolic alterations in the human erythrocyte produced by increases in glucose concentration. The role of the polyol pathway. J Clin Invest 1971;50:2105–12.Search in Google Scholar

36. Traykov TT, Jain RK. Effect of glucose and galactose on red blood cell membrane deformability. Int J Microcirc Clin Exp 1987;6:35–44.Search in Google Scholar

37. Matsuzawa T, Ikarashi Y. Haemolysis of various mammalian erythrocytes in sodium chloride, glucose and phosphate-buffer solutions. Lab Anim 1979;13:329–31.10.1258/002367779780943297Search in Google Scholar PubMed

38. Lemos GS, Márquez-Bernardes LF, Arvelos LR, Paraíso LF, Penha-Silva N. Influence of glucose concentration on the membrane stability of human erythrocytes. Cell Biochem Biophys 2011;61:531–7.10.1007/s12013-011-9235-zSearch in Google Scholar PubMed

39. Schalm OW, Jain NC, Caroll EJ. Veterinary hematology, 3rd ed. Philadelphia: Lea and Fabiger, 1975.Search in Google Scholar

40. Glaji YA, Mani AU, Igbokwe IO. Relationship of faecal egg count with packed cell volume and anaemia in Sahel sheep and goats in semi-arid northeastern Nigeria. Comp Clin Pathol 2013. doi 10.1007/s00580-013-1762-0. [Epub ahead of print 30 May 2013].10.1007/s00580-013-1762-0Search in Google Scholar

41. Ochei J, Kolhatkar A. Medical laboratory science theory and practice. New Delhi: Tata McGraw-Hill Publishing Company Limited, 2007:322.Search in Google Scholar

42. Cossins AR, Gibson JS. Volume-sensitive transport system and volume homeostasis in vertebrate red blood cells. J Exp Biol 1997;200:343–52.10.1242/jeb.200.2.343Search in Google Scholar PubMed

43. Lang F, Busch GL, Ritter M, Voèlkl H, Waldegger S, Gulbins E, et al. Functional significance of cell volume regulatory mechanisms. Physiol Rev 1998;78:247–306.10.1152/physrev.1998.78.1.247Search in Google Scholar PubMed

44. LaCelle PL, Rothstein A. The passive permeability of red blood cell to cations. J Gen Physiol 1966;50:171–88.10.1085/jgp.50.1.171Search in Google Scholar PubMed PubMed Central

45. Cueff A, Seear R, Dyrda A, Bouyer G, Egée S, Esposito A, et al. Effect of elevated intracellular calcium on the osmotic fragility of human red blood cells. Cell Calcium 2010; 47:29–36.10.1016/j.ceca.2009.11.002Search in Google Scholar PubMed

46. Davson H. Studies on the permeability of erythrocytes. VI.The effect of reducing the salt content of the medium surrounding the cell. Biochem J 1939;33:389–401.10.1042/bj0330389Search in Google Scholar

47. Zeidler RB, Kim HD. Effects of low electrolyte media on salt loss and hemolysis of mammalian red blood cells. J Cell Physiol 1979;100:551–62.10.1002/jcp.1041000317Search in Google Scholar

48. Kregenow FM. The response of duck erythrocytes to nonhemolytic hypotonic media. Evidence of a volume-controlling mechanism. J Gen Physiol 1971;58:372–95.10.1085/jgp.58.4.372Search in Google Scholar

49. Russell JM. Sodium-potassium-chloride cotransport. Physiol Rev 2000;80:211–76.10.1152/physrev.2000.80.1.211Search in Google Scholar

50. Lowenstein LM. The effect of albumin on osmotic hemolysis. Exp Cell Res 1960;20:56–65.10.1016/0014-4827(60)90221-4Search in Google Scholar

Received: 2014-2-14
Accepted: 2014-6-4
Published Online: 2014-8-5
Published in Print: 2015-3-1

©2014 by De Gruyter

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