Original Article

Treatment of Murine Toxoplasmosis with Oral and Parenteral Artemether and Following by Detection of B1 Gene by Quantitative Real-Time PCR (qPCR) for Evaluating Parasite Density

Abstract

Background: Toxoplasma gondii is an obligate intracellular parasite that can infect humans and animals. As the choice drug have shown side effects, development a new drug with low toxicity will be necessary.

Methods: BALB/c mice were infected with tachyzoiets of T. gondii. After treatment by oral and parenteral artemether (250 µg/mice) and sulfadiazine (50 µg/mice), we evaluated the rates of survival in treated and control mice. The fold change of B1 gene (target gene) expression in liver and brain of mice treated with parenteral artemether (i.p.), oral artemether (via gavage) and sulfadiazine, were detected by using the Real-Time quantitative PCR.

Results: Both treatment with sulfadiazine and artemether showed significant prolongation in time to death of the infected mice compared to the control group. Median survival days for parenteral artemether, oral artemether, sulfadiazine and control group were 8, 11, 12 and 6 d respectively. Expression of B1 gene in liver and brain of mice after treatment with artemether and sulfadiazine were reduced in comparison to housekeeping gene (β-tubulin gene). The fold change (comparing to control group) for parenteral artemether, oral artemether, sulfadiazine is 0.034, 0.027 and 0.111 for liver and 0.220, 0.425 and 0.366 for brain respectively.

Conclusion: Artemether is effective to control the tachyzoites of T. godii in vivo conditions and oral treatment is more effective than parenteral treatment. Due to its low cytotoxicity and its high effective action against the tachyzoietes of T. godii in susceptible animals.

1. Gautam B, Singh G, Wadhwa G, Farmer R, Singh S, et al. Metabolic pathway analy-sis and molecular docking analysis for identification of putative drug targets in Toxoplasma gondii: novel approach. Bioin-formation. 2012, 8(3):134-41.
2. Zhou P, Chen Z, Li HL, et al. Toxoplasma gondii infection in humans in China. Parasit Vectors. 2011; 4:165.
3. Barbosa BF, Gomes AO, Ferro EA, et al. Enrofloxacin is able to control Toxoplasma gondii infection in both in vitro and in vivo experimental models. Vet Parasitol.2012, 187(1-2):44-52 .
4. Xu X, Liu T, Zhang A, Huo X, Luo Q et al. Reactive oxygen species-triggered trophoblast apoptosis is initiated by endo-plasmic reticulum stress via activation of caspase-12, CHOP, and the JNK pathway in Toxoplasma gondii infection in mice. Infect Immun. 2012; 80(6):2121-32.
5. Lai BS, Witola WH, El Bissati K et al. Mo-lecular target validation, antimicrobial deliv-ery, and potential treatment of Toxoplasma gondii infections. Proc Natl Acad Sci USA. 2012; 109(35):14182-7.
6. Gautam B, Singh G, Singh S. Virtual screening of threonine synthase as a target for antimicrobial resistance in Toxoplasma gondii. Elixir Appl Biology. 2012; 48:9542–5.
7. Yang K, Krug EC, Marr JJ, Berens R. In-hibition of growth of Toxoplasma gondii by qinghaosu and derivatives. Antimicrob Agents Chemother. 1990; 34(10):1961-5.
8. Jones-Brando L, Angelo JD, Posner GH, Yolken RH. In vitro inhibition of Toxo-plasma gondii by four new derivatives of ar-temisinin. Antimicrob Agents Chemother. 2006; 50(12):4206-8.
9. Dunay IR, Chan WC, Haynes RK, Sibley LD. Artemisone and artemiside control acute and reactivated toxoplasmosis in a murine model. Antimicrob Agents Chemother. 2009; 53(10):4450-6.
10. 1D’Angelo JG, Bordón C, Posner GH, et al. Artemisinin derivatives inhibit Toxoplas-ma gondii in vitro at multiple steps in the lyt-ic cycle. J Antimicrob Chemother. 2009; 63(1):146-50.
11. Mesquita RT, Ziegler AP, Hiramoto RM, et al. Real-time quantitative PCR in cerebral toxoplasmosis diagnosis of Brazilian hu-man immunodeficiency virus-infected pa-tients. J Med Microbiol. 2010; 59(Pt 6):641-647.
12. Grigg ME, Boothroyd JC. Rapid identifi-cation of virulent type I strains of the pro-tozoan pathogen Toxoplasma gondii by PCR-restriction fragment length polymorphism analysis at the B1 gene. J Clin Microbiol. 2001; 39(1):398-400.
13. Buchbinder S, Blatz R, Rodloff AC; 2003 Comparison of real-time PCR detection methods for B1 and P30 genes of Toxo-plasma gondii; Diagn Microbiol Infect Dis. 2003, 45(4):269-71.
14. Mikaeiloo H, Ghaffarifar F, Dalimi A, Sharifi Z, Hassan ZM. Apoptotic activity and anti-Toxoplasma effects of artemether on the tachyzoites and experimental infect-ed Vero and J774 cell lines by Toxoplasma gondii. Indian J Pharmacol. 2016; 48(2):179-85.
15. de Oliveira T.C, Silva D A O, Rostkowsk C, et al. Toxoplasma gondii: Effects of Artemi-sia annua L. on susceptibility to infection in experimental models in vitro and in vivo. Exp Parasitol. 2009; 122(3):233-41.
16. Grujic J, Djurkovic-Djakovic O, Nikolic A, Klun I, Bobic B. Effectiveness of spiramy-cin in murine models of acute and chronic toxoplasmosis. Int J Antimicrob Agents. 2005; 25(3):226-30.
17. Ferreira RA, Oliveira AB, Ribeiro MF et al. Toxoplasma gondii: in vitro and in vivo activi-ties of the hydroxynaphthoquinone 2-hydroxy-3-(1-propen-3-phenyl)-1, 4-naphthoquinone alone or combined with sulfadiazine. Exp Parasitol. 2006; 113(2):125-9.
18. Costa JM, Darde ML, Assouline B, Vidaud M, Bretagnes S. Microsatellite in the β-tubulin gene of Toxoplasma gondii as a new genetic marker for use in direct screening of amniotic fluids. J Clin Micro-biol. 1997; 35(10):2542-5.
19. Schmittgen TD, Zakrajsek BA, Mills AG, Gorn V, Singer MJ, Reed MW. Quantita-tive reverse transcription-Polymerase Chain Reaction to study mRNA decay: comparison of endpoint and Real-Time methods. Anal Biochem. 2000; 285(2):194-204.
20. Nagamune K, Moreno SNJ, Sibley LD. Artemisinin-resistant mutants of Toxoplas-ma gondii have altered calcium homeostasis. Antimicrob Agents Chemother. 2007; 51(11):3816-23.
21. Montoya A, Miro G, Blanco MA, Fuentes I. Comparison of nested PCR and Real-Time PCR for the detection of Toxoplasma gondii in biological samples from naturally infected cats. Res Vet Sci. 2010; 89(2):212-3.
22. Sarciron NE, Sacharin C, Petavy AF, Pey-ron F. Effects of Artesunate, Dihydroar-temisinin, and an Artesunate-Dihydroartemisinin Combination Against Toxoplasma gondii. Am J Trop Med Hyg. 2000; 62(1):73-6.
23. Blum W, Pfaar U, Kuhnol J. Rapid charac-terization of artemether and its in vitro metabolites on incubation with bovine hemoglobin, rat blood and dog blood by capillary gas chromatography-chemical ionization mass spectrometry. J Chroma-togr B Biomed Sci Appl. 1998; 710(1-2):101-13.
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IssueVol 17 No 1 (2022) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v17i1.9016
Keywords
Toxoplasma gondii Artemether Oral Parenteral BALB/c mice Polymerase chain reac-tion
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How to Cite
1.
Mikaeiloo H, Ghaffarifar F, Dalimi A, Zuhair Hassan M, Sharifi Z. Treatment of Murine Toxoplasmosis with Oral and Parenteral Artemether and Following by Detection of B1 Gene by Quantitative Real-Time PCR (qPCR) for Evaluating Parasite Density. Iran J Parasitol. 2022;17(1):53-61.