Cytocidal effects of Bacillus thuringiensis crystal protein against Leishmaniatropica

Subject Areas : Medical Microbiology

Naghmeh Feridoni ¹ , Elham Moazamian ² , Manochehr Rasouli ³

1 - M.Sc., Department of Microbiology, Faculty of Science, Science and Research branch, Islamic Azad University, Fars, Iran.
2 - Assistant Professor, Department of Microbiology, Faculty of Science, Agriculture and Modern Technology, Shiraz branch, Islamic Azad University, Shiraz, Iran.
3 - Assistant Professor, Medical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

Received: 2017-06-27 Accepted : 2017-08-23 Published : 2018-08-23

Keywords: Bacillus thuringiensis, Promastigote, Leishmania tropica, Crystal protein,

Abstract :

Background & Objectives: Leishmaniasis is an infectious disease that is caused by an obligate intracellular parasite which belongs to genus Leishmania, order Kinetoplastida. Using bio-microbial control agents as one of the alternative therapies for leishmaniasis infections has been developed. The aim of this study was to assess Bacillus thuringiensis crystal protein effects against Leishmania tropica parasite.Materials & Methods: In this cross-sectional descriptive study, 47 B. thuringiensis isolates from soil samples in different regions of Fars province were collected based on the phenotypic characteristic, crystal toxin staining, and molecular identification. After enzyme digestion of toxin isolates using Proteinase K, their cytocidal effects on L. tropica were analyzed. The isolates with the most toxicity effect were identified by PCR.Results: Our results identified one isolate with the most cytocidal effect on L. tropica. The sb35 isolate crystalline toxin (Cry1) was the most effective toxin on Leishmania, showing the highest cytocidal effect (about 40%) at the concentration of 10-1 mg/ml. Cytopathic effects of promastigote treatment with crystallized toxins were observed as shrinking and inflammation.Conclusion: Considering the results obtained in this study, using crystallized proteins of B. thuringiensis can be considered as an appropriate candidate for treating leishmaniasis by performing complementary studies.

References:

1. Khan NH, Messenger LA, Wahid S, Sutherland CJ. Phylogenetic position
   of Leishmania isolates from Khyber Pakhtunkhwa province of Pakistan. Exp Parasitol. 2016;
   24(167): 61-66.
   2. Sarkari B, Ahmadpour NB, Motazedian MH, Mirjalali H, Akhoundi M, Mohebali M, Hajjaran
   H. Inter- and intraspecific variations of Leishmania strains isolated from patients with
   cutaneous and visceral leishmaniases in Fars province, south of Iran. Iran J Med Sci. 2016; 41
   (3): 209-216.
   
   3. Radosavljevic J, Naimov S. Toxicity of Bacillus thuringiensis Cry proteins against summer fruit
   tortrix (adoxophyes orana-fischer von rösslerstamm). J Invertebr Pathol. 2016; 14(138): 63-65.
   4. Sobel JD, Nyirjesy P, Brown W. Tinidazole therapy for metronidazole-resistant Vaginal
   trichomoniasis. Clin Infect Dis. 2001; 33: 1341-1346.
   5. zbi gin , arman , Kara uş , ar , oz , Kur , Ça uş , o a E, Gun uz C, an
   Gool T, Ozbel Y. Leishmaniasis in Turkey: Visceral and cutaneous leishmaniasis caused
   by Leishmania donovani in Turkey. Acta Trop. 2017; 173: 90-96.
   6. Ul Bari A. Chronology of cutaneous leishmaniasis: an overview of the history of the disease. J
   Prev Alzheimer's Dis. 2006; 16: 24-27.
   7. Lainson R. The neotropical leshmania species: a brief historical review of their discovery
   ecology and taxonomy. Rev Pan –Amaz Saude. 2010; 2: 13-32.
   8. Hanan A, El-Sadawy H, Abou El-Hag A, Janette M, Shaaban S, Hala A. In vitro activity of
   Bacillus thuringiensis (H14) 43 kDa crystal protein against Leishmania major. J Agric Environ
   Sci. 2008; 3(4): 583-589.
   9. Santana MA, Moccia CC, Gillis AE. Bacillus thuringiensis improved isolation methodology
   from soil samples. J Med Microbiol. 2008; 75: 357-358.
   10. Rampersad J. Ammons D. Bacillus thuringiensis isolation method utilizing a novel stain, low
   selection and high throughput produced atypical result. BMC Microbiol. 2005; 24(5): 52.
   11. Moazamian E, Bahador N, Rasouli M, Azarpira N. Ubiquity of parasporin producers in
   Bacillus thuringiensis natural population of Iran. Healthmed. 2012; 7(3): 971-975.
   12. Brasseur K, Auger P, Asselin E, Parent S, Cote JC, Sirois M. parasporin-2 from a new Bacillus
   thuringiensis 4r2 strain induces caspases activation and apoptosis in human cancer cells. PLoS
   One. 2015; 10(8): 135.
   13. Daneshbod Y, Oryan A, Davarmanesh M, Shirian S, Negahban S, Aledavood A, Davarpanah
   MA, Soleimanpoor H, Daneshbod K. Clinical, histopathologic, and cytologic diagnosis of
   mucosal leishmaniasis and literature review. Arch Pathol Lab Med. 2011; 135(4): 478-482.
   14. Biazar S, Moazamian E, Azarpira N. Cytocidal effects of Bacillus thuringiensis crystal protein
   on mice breast cancer cell line in in vitro condition. J Cell Tissue. 2017; 3: 243-250.
   15. Porcar M, Deleclusse A, Ibarra JE, Juarez-Perez V. Early transcription of Bacillus
   thuringiensis cry genes in strains active on lepidopteran species and the role of gene content on
   their expression. Antonie Van Leeuwenhoek. 2014; 105(6): 1007-1115.
   16. Okassov A, Nerseyan A, Kitada S, Ilin A. Parasporins as new natural anticancer agents.
   J Balkan Union Oncol. 2015; 20(1): 5-16.
   17. Armijos RX. Weigel MM. Calvopina M. Hidalgo A. Cevallos W. Correa J. Safety,
   immunogenecity, and efficacy of an autoclaved Leishmania amazonensis vaccine plus bcg
   adjuvant against new world cutaneous leishmaniasis. Vaccine. 2004; 22: 1320-1326.
   18. Moazamian E, Bahador N, Rasouli M, Azarpira N. Isolation, characterization and serotype
   classification of Bacillus thuringiensis from different soil samples of Fars province. J Microb
   World. 2010; 3: 24-30 [In Persian].
   
   19. Moazamian E, Bahador N, Rasouli M, Azarpira N, Kalani M. Investigation of cytocidal
   activity of Bacillus thuringiensis parasporal toxin on CCRF-CEM cell line. J Fasa
   Univ Med Sci. 2012; 2(4): 247-253.
   20. Poornima K, Selvanayagam P, Shenbagarathai R. Identification of native Bacillus
   thuringiensis strain from South India having specific cytocidal activity against cancer cells. J
   Appl Microbiol. 2010; 109: 348-354.
   21. Mizuki E, Park YS, Saitoh H, Yamashita S, Akao T, Higuchi K, Ohba M. Parasporin, a human
   leukemic cell-recognizing parasporal protein of Bacillus thuringiensis. Clin Diagn Lab
   Immunol. 2000; 7(4): 625- 634.
   22. Park HW, Bideshi DK, Federici BA. Molecular genetic manipulation of truncated Cry 1c
   protein synthesis in Bacillus thuringiensis to improve stability and yield. Appl
   Environ Microbiol. 2000; 66: 4449-4455.
   23. Kondo S, Mizuki E, Akao T, Ohba M. Antitrichomonal strains of Bacillus thuringiensis.
   Parasitol Res. 2002; 88: 1090-1092.
   24. Cappello M, Bungiro RD, Harrison LM. A purified Bacillus thuringiensis crystal protein with
   therapeutic activity against the hookworm parasite Ancylostoma ceylanicum. Proc Natl Acad
   Sci USA. 2006; 103: 15154-15159.
   25. Desjeux P. Leishmaniasis: current situation and new perspectives. Europ J Clin Microbiol
   Infect Dis. 2004; 27: 305-318