Montero GT, Hurtado de Mendoza AJ

Language: Spanish
References: 13
Page: 21-28
PDF size: 402.69 Kb.

ABSTRACT

Introduction: experimental methods are valuable tools for studying human biological processes. It is necessary to find the proper method of euthanasia to be used for obtaining the lowest possible variations of tissue.
Objective: to identify the presence of histopathological lesions related to euthanasia method, in the search of suitable one for the study of multiple organ failure.
Methods: BALB/c, female mice weighing 20 ± 2 g, aged 12 weeks. They were included into three groups corresponding to the methods of traction and cervical dislocation, immersion in liquid nitrogen for 7 s and over anesthesia with pentobarbital 60 mg/kg. Autopsy was performed immediately on the spot, and fragments of various organs were obtained, fixed in 10% formalin. These fragments were processed for inclusion in paraffin, sectioned and stained with hematoxylin/eosin.
Results: lesions found in the lung between focal and partial were important aspects. Cerebral edema occurred in brain dislocation and over anesthesia groups. Immersion in liquid nitrogen for 7 s afforded instantaneous death nod obtaining fragment for histopathological studies with minimum alterations.
Conclusions: the histopathological examination of the various methods of euthanasia showed that immersion in liquid nitrogen had lower postmortem changes, making its use suitable in studies which require assessing histopathological changes from slight changes in different organs lesions. It is therefore advisable for studies of systemic inflammatory response in experimental models.

REFERENCES

1. Cotran RS, Kumar V, Collins. Patología Estructural y Funcional de Robbins. Madrid: Mc Graw-Hill; Interamericana; 2005. p. 5.

2. Fiette L, Slaoui M. Necropsy and sampling procedures in rodents. Methods Mol Biol. 2011;691:39-67.

3. Hurtado de Mendoza J, Zorrilla JF. Elaboración de un modelo experimental de quemaduras. Experiencias preliminares. Rev Cubana Med Milit. 1982;1(2):79-89.

4. Roch-Arveillerm, Fontagne J, Abbouyi EL, Ralchvarg D, Giroud JP. Effect of an immunomodulating agent, RU 41740, on polymorphonuclear responsiveness after burn injury. Inflammation. 1991;15(6):137-41.

5. Nakanishi T, Nishi Y, Sato EF, Ishii M, Hamada T, Inove M. Thermal injury induces Thymocyte apoptosis in the rat. J Trauma. 1998;44(1):143-8.

6. Ramos N, Lorente JA. Modelos experimentales de sepsis. Revista Electrónica de Medicina Intensiva. Artículo nº C34; 2005;5(5). Disponible en: http://remi.uninet.edu/2005/05/REMIC34.pdf

7. Esmon CT. Why do animal models of sepsis (sometimes) fail to mimic human sepsis. Crit Care Med. 2004;32:219-22.

8. Dewals BG, Vanderplasschen A. Malignant catarrhal fever induced by Alcelaphine herpesvirus 1 is characterized by an expansion of activated CD3+CD8+CD4- T cells expressing a cytotoxic phenotype in both lymphoid and non-lymphoid tissues. Vet Res. 2011;42(1):95.

9. Montero T, Hurtado de Mendoza J. Preguntas y respuestas sobre el daño múltiple de órganos. Reflexiones acerca de la temática. Rev Cubana Med Mil. 2010;39(1)44-56.

10. Couto M. Laboratory guidelines for animal care. Methods Mol Biol. 2011;770:579-99.

11. Ohno Y. Importance of research on alternatives to animal experiments in pharmaceutical sciences. Yakugaku Zasshi. 2008;128(5):735-40.

12. Howell RL, Donegan CL, Pinkert CA. Mouse embryo yield and viability after euthanasia by CO2 inhalation or cervical dislocation. Comp Med. 2003;53(5):510-3.

13. Sáenz Medina J, Asuero de Lis MS, Correa Gorospe C, Cuevas B, Gómez Dos Santos V, et al. Modelos experimentales para la investigación y el entrenamiento en trasplante renal. Actas Urol Esp. 2008;32(1):83-90.