Study of hemolytic activity of body extracts Ectomyelois ceratoniae (Lepidoptera: Pyralidae)

Subject Areas : Developmental biology of plants and animals , development and differentiation in microorganisms

isa jabaleh ¹ , majid kazzazi ² , ahmad asoodeh ³

1 - Department of Plant Protection, College of Agriculture, Bu–Ali Sina University, Hamedan, Iran.
2 - 1. Department of Plant Protection, College of Agriculture, Bu–Ali Sina University, Hamedan, Iran.
3 - Department of Chemistry and Biochemistry, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran.

Received: 2017-01-07 Accepted : 2017-03-07 Published : 2017-02-19

Keywords: blood, Ectomyelois ceratoniae, Hemolysis Assay,

Abstract :

Antimicrobial peptides (AMPs) have attracted attention as a novel class of antimicrobial agents because AMPs efficiently kill a wide range of species, including bacteria, fungi, and viruses, via a novel mechanism of action. One representative problem is the hemolytic activity of AMPs. Hemolysis is the premature destruction of erythrocytes. The hemolysis assay is used to determine the hemolytic effect of a test compound. Hemolytic activity should be investigated in the process of identifying antimicrobial peptides of insects and future therapeutic applications. In this study, for hemolytic activity of the body extracts larvae Ectomyelois ceratoniae (Zeller), 5 ml of fresh human, sheep and hen blood was added to a heparinized tube. The hemolytic assay was then carried out according to the methods reported in previous studies. The absorbance at 567 nm was then measured. Triton X-100 (0.1%), which yields 100% hemolysis, was used as a positive control. The results were compared with the control. Human and sheep red blood cells showed good tolerance while exposed to the body extracts larvae E. ceratoniae. The body extracts had no significant hemolytic effect in the concentration range (0.5–2 mg/ml) against human and sheep blood, but had a little hemolytic effect against hen blood. The results showed that hemolysis activity varies according to blood type. Also, the body extracts larvae E. ceratoniae did not have a hemolytic effect on red blood cells. As a result, it can be a good candidate as an antibiotic in complementary studies.

References:

[1] Asoodeh, A., Zardini, H.Z. and Chamani, J., (2012). Identification and characterization of two novel antimicrobial peptides, temporinRa and temporin‐Rb, from skin secretions of the marsh frog (Rana ridibunda). Journal of Peptide Science, 18(1), pp.10-16.

[2] Bulet, P., Stöcklin, R., & Menin, L. (2004). Anti‐microbial peptides: from invertebrates to vertebrates. Immunological reviews, 198 (1), 169-184.

[3] Chou, H. T., Kuo, T. Y., Chiang, J. C., Pei, M. J., Yang, W. T., Yu, H. C., ... & Chen, W. J. (2008). Design and synthesis of cationic antimicrobial peptides with improved activity and selectivity against Vibrio spp. International journal of antimicrobial agents, 32 (2), 130-138.

[4] Dathe, M., & Wieprecht, T. (1999). Structural features of helical antimicrobial peptides: their potential to modulate activity on model membranes and biological cells. Biochimica et Biophysica Acta (BBA)Biomembranes. 1462 (1), 71-87.

[5] Einhorn, T. A. (1998). The cell and molecular biology of fracture healing. Clinical orthopaedics and related research, 355, S7-S21.

[6] Hancock, R. E., & Sahl, H. G. (2006). Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nature biotechnology, 24 (12), 1551.

[7] Helmerhorst, E. J., Reijnders, I. M., van't Hof, W., Veerman, E. C., & Nieuw Amerongen, A. V. (1999). A critical comparison of the hemolytic and fungicidal activities of cationic antimicrobial peptides. FEBS letters, 449 (2-3), 105-110.

[8] Horie, T., Sugiyama, Y., Awazu, S., & Hanano, M. (1981). The correlation between drug binding to the human erythrocyte and its hemolytic activity. Journal of pharmacobiodynamics, 4 (2), 116-122.

[9] Jenssen, H., Hamill, P., & Hancock, R. E. (2006). Peptide antimicrobial agents. Clinical microbiology reviews, 19(3), 491-511.

[10] Litman, G. W., Litman, R. T., & Henry, C. J. (1976). Analysis of lipophilic carcinogenmembrane interactions using a model human erythrocyte membrane system. Cancer research, 36 (2 Part 1), 438-444.

[11] Madigan, M. T., Martinko, J. M., & Parker, J. (1997). Brock biology of microorganisms (Vol. 11). Upper Saddle River, NJ: Prentice hall.

[12] Malmsten, M., Kasetty, G., Pasupuleti, M., Alenfall, J., & Schmidtchen, A. (2011). Highly selective end-tagged antimicrobial peptides derived from PRELP. PloS one, 6 (1), e16400.

[13] Matsuzaki, K., Sugishita, K., Fujii, N., & Miyajima, K. (1995). Molecular basis for membrane selectivity of an antimicrobial peptide, magainin 2. Biochemistry, 34 (10), 3423-3429.

[14] Memarpoor-Yazdi, M., Zare-Zardini, H. and Asoodeh, A., (2013). A novel antimicrobial peptide derived from the insect Paederus dermatitis. International Journal of Peptide Research and Therapeutics, 19 (2), pp.99108.

[15] Mohammed, Kh.A., Toson, M.A. & Hassanien, H.H.M. (2010). Effects of phytase supplementation on performance and egg quality of laying hens fed diets containing rice bran. Egyptian Journal of Poultry Science, 30, 649-659. 15.

[16] Mohsinzadeh, M., Nobakht, A. & Safamehr, A. R. (2013). Effects of different levels of crude protein and probiotic (Protexin) on performance and blood metabolites of laying hens. Animal Science Journal (Pajouhesh& Sazandegi), 103, 133-144.

[17] Oyedapo, O. O., & Famurewa, A. J. (1995). Antiprotease and membrane stabilizing activities of extracts of Fagara zanthoxyloides, Olax subscorpioides and Tetrapleura tetraptera. International Journal of Pharmacognosy, 33 (1), 65-69.

[18] Robertis FA, Robertis EMH (1995). Cell and molecular biology in: cell membrance Saunders London, pp. 239-245. 

[19] Rosenthal, A. Z., & Elowitz, M. B. (2012). Following evolution of bacterial antibiotic resistance in real time. Nature genetics, 44 (1), 11-13.

[20] Schaufuss, P. and Steller, U., (2003). Haemolytic activities of Trichophyton species. Medical mycology, 41 (6), pp.511516.

[21] Sessa, G., & Weissmann, G. (1968). Effects of four components of the polyene antibiotic, filipin, on phospholipid spherules (liposomes) and erythrocytes. Journal of Biological Chemistry, 243 (16), 4364-4371.

[22] Thomas, A., Kohler, M., Mester, J., Geyer, H., Schänzer, W., Petrou, M., & Thevis, M. (2010). Identification of the growthhormone‐releasing peptide‐2 (GHRP‐2) in a nutritional supplement. Drug testing and analysis, 2 (3), 144-148.

[23] Torrent, M., Andreu, D., Nogués, V. M., & Boix, E. (2011). Connecting peptide physicochemical and antimicrobial properties by a rational prediction model. PloS one, 6 (2), e16968.

[24] Vale, P. F., Fenton, A., & Brown, S. P. (2014). Limiting damage during infection: lessons from infection tolerance for novel therapeutics. PLoS biology, 12 (1), e1001769.

[25] Warner, R. L. (1988). Contributions to the biology and the management of the carob moth, Ectomyelois ceratoniae (Zeller) in ÔDeglet NoorÕ date gardens in the Coachella Valley of California. PhD dissertation, University of California, Riverside. CA.

[26] Wieprecht, T., Dathe, M., Beyermann, M., Krause, E., Maloy, W. L., MacDonald, D. L., & Bienert, M. (1997). Peptide hydrophobicity controls the activity and selectivity of magainin 2 amide in interaction with membranes. Biochemistry, 36 (20), 6124-6132.

[27] Zibaee, A., Bandani, A. Talaei, R. & Malagoli, D. (2011). Cellular immune reactions of the sunn pest, Eurygaster integriceps, to the entomopathogenic fungus, Beauveria bassiana and its secondary metabolites. Journal of Insect Science, 11: 138: 1-16.