Toxoplasma gondii RH Ankara suşu ile intraperitoneal olarak enfekte edilen farelerde sistemik doku tutulumunun değerlendirilmesi

Abstract

Amaç: Bu çalışmada, intraperitoneal yolla Toxoplasma gondii RH Ankara suşu inoküle edilen farelerde oluşan akut tokzoplazmozisde sistemik doku tutulumunun patolojik olarak değerlendirilmesi amaçlanmıştır. Yöntem: Mus musculus varyete albino cinsi 42 adet fare, herhangi bir inokülasyonun yapılmadığı kontrol grubu, intraperitoneal inokülasyondan sonra sakrifiye edilmeyen ve intraperitoneal inokülasyondan ikinci gün ve dördüncü gün sakrifiye edilenler olmak üzere dört gruba ayrılmıştır. Farelerin intraperitoneal inokülasyonunda T. gondii RH Ankara suşunun farelerdeki 48-50 saatlik intraperitoneal pasajları kullanılmış ve her bir fareye 5x104 takizoit verilmiştir. Sakrifiye edilen hayvanlar makroskobik olarak incelendikten sonra alınan doku örnekleri %10 formalinde tespit edilmiştir. Parafin bloklara alınan dokulardan hazırlanan preparatlar hematoksilen-eozin ile boyanarak histopatolojik olarak incelenmiştir. Bulgular: Karaciğer, dalak, ince bağırsaklar, böbreklerin parankimi ve serozasında nekrozlar ve çok sayıda serbest takizoit grupları gözlenmiştir. Akciğerin alveolar kapiller lümenlerinde de takizoitler saptanmıştır. Beyin ve beyincikte ise herhangi anlamlı bir patolojik değişiklik izlenmemiştir. Dokulardaki nekroz ve Toxoplasma gondii takizoidlerinin yoğunluğunun, inokülasyon süresi ve temas yüzeyi ile doğru orantılı olduğu gözlenmiştir. Herhangi bir müdahalede bulunulmayan enfekte farelerin tamamı altıncı günün sonunda ölmüştür. Bu çalışmada T. gondii RH Ankara suşunun farelerde virulan Tip I gruba bağlı RH suşlarındakine benzer patolojik bulgular oluşturduğu belirlenmiştir. Sonuç: Fare inokülasyonu tanı amacıyla kullanılabileceği gibi aşı, ilaç ve patogeneze yönelik çalışmalarda da bu modelden yararlanılabilir

Keywords

• Toxoplasma gondii, patogenez, fare

Evaluation of systemic tissue involvement in mice following intraperitoneal inoculation of Toxoplasma gondii RH Ankara strain

Abstract

Objective: This study aimed to pathologically evaluate systemic tissue involvement in acute toxoplasmosis in mice inoculated with Toxoplasma gondii RH Ankara strain. Method: Forty-two Mus musculus albino mice were divided into four groups with the control group receiving no inoculation, the group consisting of mice that were not sacrificed after intraperitoneal inoculation and the groups consisting of mice euthanasia were performed on day runner up or day fourth following intraperitoneal inoculation. A 48-50-hour intraperitoneal passage of T. gondii RH Ankara strain in mice was used for inoculation and each mouse received 5x104 tachyzoites. The mice sacrificed by euthanasia were macroscopically examined and then collected tissue samples fixed in 10% formalin. The preparations set by from the tissues blocked in paraffin were stained with hematoxylin-eosin and histopathological examination was performed. Results: Necrosis in the serosa of the intestine and parenchyma of the liver, spleen, kidneys, pancreases and omentum and many free groups of tachyzoites were observed. Tachyzoites were identified in alveolar capillary lumens in the lungs. No pathological finding was observed in the cerebrum and cerebellum but few tachyzoites. It was observed that tissue necrosis and the density of Toxoplasma gondii tachyzoites were directly proportional to the duration of post inoculation. All infected mice that received no intervention died on day sixth. In this study in mice, it was found that T. gondii RH Ankara strain induced the same pathological findings as the RH strain belonging to the virulent Type I group. Conclusion: Mice inoculation can be used for diagnosis and this model may also be used in studies investigating vaccination, drugs and pathogenesis.

Keywords

• Toxoplasma gondii, pathogenesis, mice

References

1. 1. Sukthana Y, Waree P, Pongponratn E, Chaisri U, Riganti M. Pathologic study of acute toxoplasmosis in experimental animals. Southeast Asian J Trop Med Public Health, 2003; 34(1): 16-21.
2. 2. Ozkan AT, Celebi B, Babür C, Lucio-Forster A, Bowman DD, Lindsay DS. Investigation of antiToxoplasma gondii antibodies in cats of the Ankara region of Turkey using the Sabin-Feldman dye test and an indirect fluorescent antibody test. J Parasitol, 2008; 94(4): 817-20.
3. 3. Dubey JP. Toxoplasma gondii in medical microbiology Baron S, ed. 4th edition. Galveston (TX): University of Texas Medical Branch at Galveston, 1996.
4. 4. Buffalano W, Beghetto E, Del Pezzo M, Spadoni A, Cristina M, Petersen E, et al. Use of recombinant antigens for early postnatal diagnosis of congenital toxoplasmosis. J Clin Microbiol, 2005; 43: 5916-24.
5. 5. Haholu A, Yıldırım S, Koru O, Ardıç N, Baloglu H. Toksoplazma gondii lenfadenitinin sitolojik tanısı: Olgu sunumu. Türk Patoloji Dergisi, 2006; 22(1): 57-60.
6. 6. Piergili Fioretti D. Problems and limitations of conventional and innovative methods for the diagnosis of toxoplasmosis in humans and animals. Parasitologia, 2004; 46(1-2): 177-81.
7. 7. Döskaya M, Degirmenci A, Ciçek C, Ak M, Korkmaz M, Gürüz Y, et al. Behaviour of Toxoplasma gondii RH Ankara strain tachyzoites during continuous production in various cell lines. Parasitology, 2006; 132(Pt 3): 315-9.
8. 8. Djurković-Djaković O, Nikolić A, Bobić B, Klun I, Aleksić A. Stage conversion of Toxoplasma gondii RH parasites in mice by treatment with atovaquone and pyrolidine dithiocarbamate. Microbes Infect, 2005; 7(1): 49-54.

9. 9. Araujo FG, Remington JS. Effect of clindamycin on acute and chronic toxoplasmosis in mice. Antimicrob Agents Chemother, 1974; 5(6): 647-51.
10. 10. Delibas SB, Ertabaklar H, Ertug S. Evaluation of antigenic variations between two virulent Toxoplasma strains. J Medical Microbiol, 2006; 55: 1333-5.
11. 11. Zenner L, Foulet A, Caudrelier Y, Darcy F, Gosselin B, Capron A, et al. Infection with Toxoplasma gondii RH and prugniaud strains in mice, rats and nude rats: kinetics of infection in blood and tissues related to pathology in acute and chronic infection. Pathol Res Pract, 1999; 195(7): 475-85.
12. 12. Sibley LD, Mordue D, Howe DK. Experimental approaches to understanding virulence in toxoplasmosis. Immunobiology, 1999; 201(2): 210-24.
13. 13. Ekmen H, Altıntaş K. Bir köpekten Toxoplasma izolmanı. Turk Hij Tec Biol Derg, 1973; 33: 17-20.
14. 14. Hazıroglu R, Altıntas K, Atasever A, Gulbahar MY, Kul O, Tunca R. Pathological and immunohistochemical studies in rabbits experimentally infected with Toxoplasma gondii. Turk J Vet Anim Sci, 2003; 27:285-93.
15. 15. Yaman K, Aral Akarsu G, Güngör C, Ataoğlu H. Toxoplasma gondii’nin çeşitli suşlarının proteinlerinin karakterizasyonu. Turk Parazitol Derg, 2011; 35: 133-6.
16. 16. Yaman K. Toxoplasma gondii antijenlerinin karakterizasyonu. Yüksek Lisans Tezi, Ankara Üniversitesi Sağlık Bilimleri Enstitüsü, 2007.
17. 17. Elwell MR, Frenkel JK. Acute toxoplasmosis in hamsters and mice: measurement of pathogenicity by fever and weight loss. Am J Vet Res, 1984; 45(12): 2663-7.
18. 18. Csóka R, Császár S. The lymphoid system of Toxoplasma infected mice. Acta Microbiol Hung, 1983; 30(1): 43-6.
19. 19. Djurković-Djaković O, Klun I, Khan A, Nikolić A, Knezević-Usaj S, Bobić B, et al. A human origin type II strain of Toxoplasma gondii causing severe encephalitis in mice. Microbes Infect, 2006; 8(8): 2206-12.
20. 20. Eissa MH, Antonious SN, Salama MM, Fikry AA, Morsy TA. Histopathological studies of acute, chronic and congenital infections of toxoplasmosis in mice. J Egypt Soc Parasitol, 1990; 20(2): 805-16.
21. 21. Degerli K, Kilimcioglu AA, Kurt O, Tamay AT, Ozbilgin A. Efficacy of azithromycin in a murine toxoplasmosis model, employing a Toxoplasma gondii strain from Turkey. Acta Trop, 2003; 88: 45–50.
22. 22. Waree P, Ferguson DJ, Pongponratn E, Chaisri U, Sukthana Y. Immunohistochemical study of acute and chronic toxoplasmosis in experimentally infected mice. Southeast Asian J Trop Med Public Health, 2007; 38(2): 223-31.
23. 23. De Champs C, Pelloux H, Dechelotte P, Giraud JC, Bally N, Ambroise-Thomas P. Toxoplasma gondii infection in rats by the RH strain: inoculum and age effects. Parasite, 1998; 5(3): 215-8.
24. 24. Armstrong JA, Fulton JD. Observations on the pathology of toxoplasmosis in the cotton rat. Br J Exp Pathol, 1959; 40(3): 225-31.