A modified paperfuge technique in evaluating anemia in phenylhydydrazine-induced ICR
Anemia is a common observation in several diseases in humans and animals. This condition is evaluated through the blood using parameters, including the packed cell volume (PCV). However, quick diagnosis might take longer, especially in resource limited areas, as blood samples still need to be processed in the laboratory. In this experimental study, a modified paperfuge technique was evaluated to assess anemia in an animal model. A total of 16 ICR mice were used. After animal acclimatization for ten days, persistent anemia (hemolytic) was induced in nine mice by administering phenylhydrazine every other day for three times. Only distilled water was used in the control mice. After seven days, blood was extracted from the mice. Complete blood count using a species specific hematology machine was performed. PCV was further evaluated using the centrifuge machine and the modified paperfuge technique. Results using the hematology machine showed that the phenylhydrazine-induced mice had lower red blood cell count (x̄= 4.26 x 10^6/µL, SD=0.93) and PCV (x̄=32.5, SD=7.5) than the red blood cell count (x̄=5.26 x 10^6/µL, SD=1.1) and PCV (x̄=36.5, SD=5.8) of the control mice. PCV readings from the three methods were strongly correlated with the RBC count. On the other hand, PCV readings from the paperfuge technique were also strongly and significantly correlated with the centrifuge (R=0.99, p=0.000) and hematology machine (R=0.89, p=0.000) results. Further analysis revealed that there was no significant difference observed between the PCV readings of the CBC machine, the centrifuge and the modified paperfuge (treatment group: p=0.39, control group: p=0.09). Results suggest that the modified paperfuge technique can be reliable in measuring PCV to evaluate anemia. Further studies are recommended to compare results in anemic animals and humans.
2. Bekkevold, C. M., Robertson, K. L., Reinhard, M. K., Battles, A. H., & Rowland, N. E. (2013). Dehydration parameters and standards for laboratory mice. Journal of the American Association for Laboratory Animal Science, 52(3), 233-239.
3. Bhamla, M. S., Benson, B., Chai, C., Katsikis, G., Johri, A., & Prakash, M. (2017). Hand-powered ultra-low cost paper centrifuge. Nature Biomedical Engineering, 1(1), 0009.
4. Bhamla, M. S., Benson, B., Chai, C., Katsikis, G., Johri, A., & Prakash, M. (2017). Paperfuge: An ultralow-cost, hand-powered paper-centrifuge inspired by the mechanics of a whirligig toy. Bulletin of the American Physical Society, 62.
5. Bhamla, M. S., Benson, B., Chai, C., Katsikis, G., Johri, A. & Prakash, M. (2016). Paperfuge: An ultra-low cost, hand-powered centrifuge inspired by the mechanics of a whirligig toy. bioRxiv, 072207. doi: https://doi.org/10.1101/072207
6. Davis, T. M., Krishna, S., Looareesuwan, S., Supanaranond, W., Pukrittayakamee, S., Attatamsoonthorn, K. & White, N. J. (1990). Erythrocyte sequestration and anemia in severe falciparum malaria. Analysis of acute changes in venous hematocrit using a simple mathematical model. Journal of Clinical Investigation, 86(3), 793-800.
7. Foltz, C. J., & Ullman-Cullere, M. (1999). Guidelines for assessing the health and condition of mice. Lab Animal, 28(4), 28-32
8. Gebretsadkan, T. K., Ambachew, G., & Birhaneselassie, H. (2015). The comparison between microhematocrit and automated methods for hematocrit determination. International Journal Blood Research and Disorders 2(1), 1-3.
9. Goodnough, L. T., Brecher, M. E., Kanter, M. H., & AuBuchon, J. P. (1999). Transfusion medicine—blood transfusion. New England Journal of Medicine, 340(6), 438-447.
10. Grimm, R. H., Neaton, J. D. & Ludwig, W. (1985). Prognostic importance of the white blood cell count for coronary, cancer, and all-cause mortality. Journal of the American Medical Association, 254(14), 1932-1937.
11. Hara, H., & Ogawa, M. (1976). Erythropoietic precursors in mice with phenylhydrazine-induced anemia. American Journal of Hematology, 1(4), 453-458.
12. Kularatne, S. A. M., Gawarammana, I. B., & Kumarasiri, P. R. V. (2005). Epidemiology, clinical features, laboratory investigations and early diagnosis of dengue fever in adults: A descriptive study in Sri Lanka. Southeast Asian Journal of Tropical Medicine and Public Health, 36(3), 686-692.
13. Myers, G. J. & Browne, J. (2007). Point of care hematocrit and hemoglobin in cardiac surgery: A review. Perfusion, 22(3), 179-183.
14. Paul, A., Calleja, L., Vilella, E., Martı́nez, R., Osada, J., & Joven, J. (1999). Reduced progression of atherosclerosis in apolipoprotein E-deficient mice with phenylhydrazine-induced anemia. Atherosclerosis, 147(1), 61-68.
15. Purves, W. K., Sadava, D. E., Orians, G. H., & Craig, H. H. (2004). Life: The Science of Biology (7th ed.). Sunderland, Mass: Sinauer Associates.
16. Rosman, N. P. (2003). Evaluation of the child who convulses with fever. Pediatric Drugs, 5(7), 457-461.
17. Spivak, J. L., Toretti, D., & Dickerman, H. W. (1973). Effect of phenylhydrazine-induced hemolytic anemia on nuclear RNA polymerase activity of the mouse spleen. Blood, 42(2), 257-266.
18. Stockham, S. L., & Scott, M. A. (2013). Fundamentals of veterinary clinical pathology. John Wiley & Sons.
19. Ullman-Culleré, M. H., & Foltz, C. J. (1999). Body condition scoring: a rapid and accurate method for assessing health status in mice. Comparative Medicine, 49(3), 319-323.
20. Uthman, E. (2009). Understanding Anemia. Univ. Press of Mississippi. ISBN 9781604737011.
21. Valentine, W. N., & Paglia, D. E. (1984). Erythrocyte enzymopathies, hemolytic anemia, and multisystem disease: an annotated review. Blood, 64(3), 583-591.
22. Ybañez, A. P., Ybañez, R. H. D., Villavelez, R. R., Malingin, H. P. F., Barrameda, D. N. M., Naquila, S. V., & Olimpos, S. M. B. (2016). Retrospective analyses of dogs found serologically positive for Ehrlichia canis in Cebu, Philippines from 2003 to 2014. Veterinary World, 9(1), 43-47.
Copyright (c) 2017 University of the Visayas
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.