Potential Use of Some Liquid Natural Media for the Production of Blastospores of Entomopathogenic Fungi Metarhizium anisopliae and Beauveria bassiana

Document Type : Research Paper


1 Ph.D. student, Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran

2 Professor, Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran

3 Professor, Department of Food Science and Technology, Faculty of Agricultural Engineering and Technology, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran

4 Research Scientist, Lethbridge Research Center, Lethbridge, Alberta, Canada


In the present study, the potential of natural ingredients such as extract of pug, carrot pulp, permeate and fish waste were investigated to produce blastospores of two fungi Beauveria bassiana and Metarhizium anisopliae. SDB and PDB culture media were considered as control. In the case of absence of the initial pH for culture media, the highest production of blastospores was recorded in SDB medium and permeate extract for B. bassiana and M. anisopliae respectively, in three days. In the second case, with the same initial pH (5.6) for culture media, the maximum production of blastospores was observed in the extract of fish waste (3 × 107 blastospore /ml medium) and in the carrot pulp extract (6 × 106 blastospore /ml medium) for B. bassiana and M. anisopliae respectively. In the next step, wheat bran and permeate (4% and 8%) extracts were added to selected medium fish waste and carrot pulp. Addition of extract of permeate (8%) had the greatest effect on increasing blastospor production. In the last step, mixed different salts were added and reduced blastospore production in fungus B. bassiana while increased in M. anisopliae compared to the control medium. Finally, two optimum media were introduced for the production of two entomopathogenic fungi.


Main Subjects

Asgari, H.,  Zamani, S.M. and Ashouri, A. 2009. Effect of Some Liquid and Solid Media on Sporulation of Beauveria bassiana. Iranian Journal of Plant Protection Science 39 (1): 31-43 (In Farsi).
AOAC (1990). Official methods of analysis, 15th Edition. Association of Official Analytical Chemists, Washington DC, USA. pp: 554,575,654.
Balakrishnan, S., Srikanth, J., Santhalakshmi, G., Hari, K. & Sankaranarayanan, C. (2011). Response of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae to molasses media fortified with supplements. Journal of Sugarcane Research, 1(2), 57-65.
Bena-Molaei, P., Talaei-Hassanloui, R., Askary, H. & Kharazi-Pakdel, A. (2009). Study on potential of some solid natural substances in production of Beauveria bassiana (Ascomycota: Cordycipitaceae) conidia. Journal of Entomological Society of Iran, 30 (2), 1-15. (In Persian).    
 Bena-Molaei, P., Talaei-Hassanloui, R. & Askary, H. (2011). Effect of culture substrates on virulence of Beauveria bassiana (Ascomycota: Cordycipitaceae) conidia against the browntail moth, Euproctis chrysorrhoea (Lep.: Lymantriidae). Journal of Biocontrol Science and Technology, 21(5), 619- 624.
Bigham, Z. and Talaei-Hassanloui, R. 2017. Effect of Some Liquid and Solid Media on Sporulation of Beauveria bassiana. Biological Control pf Pests and Plant Diseases 6 (1): 103-109 (In Farsi).
Fargues, J., Smits, N., Vidal, C., Vey, A., Vega, F., Mercadier, G. & Quimby, P. (2002). Effect of liquid culture media on morphology, growth, propagule production and pathogenic activity of the hyphomycete, Metarhizium flavoviride. Journal of Mycopathologia, 154, 127-138.                                                                                                      
Feng, M. G., Poprawski, T. J. & Khachatourians, G. G. (1994). Production, formulation and application of the entomopathogenic fungus Beauveria bassiana for insect control: current status. Journal of Biocontrol Science and Technology, 4, 3-34.                                                                                                                                                               
Fransisco, E. A., Mochi, D. A., Coreia, A. C. B. & Monteiro, A. C. (2006). Influence of culture media in viability test of conidia of entomopathogenic fungi. Journal of Ciencia Rural, 36, 1309-1312.
Ibrahim, Y. B., Lim, T. K., Tang, M. K. & Teng, H. M. (1993). Influence of temperature, pH and selected growth media on germination, growth and sporulation of Aschersonia placenta and Hypocrella raciborskii. Journal of Biocontrol Science and Technology, 3, 55-61.                                                                                                                      
Jackson, M. A. (1997). Optimizing nutritional conditions for the liquid culture production of effective fungal biological control agents. Journal of Industrial Microbiology & Biotechnology, 19, 180-187.
Kamp, A. M. & Bidochka, M. J. (2002). Conidium production by insect pathogenic fungi on commercially available agar. Journal of Letters in Applied Microbiology, 35, 74-77.    
Kamyab, A. (2001). A user guide to the animal nutrition. 218 pp. Hage- Shenas publication. 
Kassa, A., Brownbridge, M., Parker, B. L., Skinner, M., Gouli, V., Gouli, S., Gou, M., Lee, F. & Hata, T. (2008). Whey for mass production of Beauveria bassiana and Metarhizium anisopliae. Journal of Mycological Research, 112 (5), 583-591.                                                                                                                                                                            
Kim, J. S., Skinner, M. & Parker, B. L. (2010). Influence of whey permeate and millet as substrates on thermotolerance of Beauveria bassiana and Metarhizium anisopliae conidia during storage. Journal of Biocontrol Science and Technology, 20(8), 859-863.                                                                                                                           
Lacey, L. A., Kirk, A. A., Millar, L., Mercadier, G. & Vidal, C. (1999). Ovicidal and larvicidal activity of conidia and blastospores of Paecilomyces fumosoroseus (Deuteromycotina: hyphomycetes) against Bemisia argentifolii (Homoptera: Aleyrodidae) with a description of a bioassay system allowing prolonged survival of control insects. Journal of Biocontrol Science and Technology, 9, 9-18. 
Li, D. P. & Holdom, D. G. (1995). Effects of nutrients on colony formation, growth, and sporulation of Metarhizium anisopliae (Deuteromycotina: Hyphomycetes). Journal of Invertebrate Pathology, 65, 253–260.تعریف‌های Beauveria bassiana (Balsamo) Vuellemin and Metarhizium anisopliae (Metschnikoff) Sorokin are entomopathogenic fungi (EPF) that occur naturally in the environment and have a potential as biological control agents against many insect pests. Different solid substrates i.e such as grains, vegetable wastes, maize, bran, cotton seed, rice husk, wheat and liquid media such as coconut water were evaluated at variable moisture content and yeast extract concentration ffor mass production of two entomopathogenic fungi: Beauveria bassiana (Bals.) Vuellemin and Metarhizium anisopliae. In the present study rice is used as a best solid substrate for spore production and their viability but fungus also grows equally well on maize or other grains. This gives an overview of the mass production of fungal spores using diphasic liquid-solid fermentation i.e LUBILOSA system which is particularly well adapted to the production of EPF’s B. bassiana and M. anisoliae and provides explanations for the use of various techniques and procedures.                                                                                                      
Ottati-de-Lima, E. L., Batista Filho, A., Almeida, J. E., Gassen, M. H., Wenzel, I. M., Almeida, A. M. & Zapellini, L. O.  (2014). Liquid production of entomopathogenic fungi and ultraviolet radiation and temperature effects on produced propagules. Journal of Arquivos do Instituto Biológico, São Paulo, 81(4), 342- 350.                                                                                                                                                                                   
Powel, K. A. (1995). The production of chemicals by biological control agents. Journal of Pesticide Science, 44,395-397.
Prasad, C. S. & Pal, R. (2014). Mass production and economics of entomopathogenic fungus, Beauveria bassiana, Metarhizium anisopliae and Verticillium lecanii on agricultural and industrial waste. Scholars Journal of Agriculture and Veterinary Sciences, 1(1), 28-32.                                                                                                                                 
Rayati, D. J., Nyoman, I., Aryantha, P.  & Arbianto, P. (2001). The optimization of nutrient factors in spore production of Paecilomyces fumosoroseus (Wize) Brown & Smith with submerged – surface fermentation system.The Fifth Symposium on Agri-Bioche, March-11-2001. Tokyo, Japan.
Shelton, A. M., Vandenberg, J. D., Ramos, M. & Wilsey, W. T. (1998). Efficacy and persistence Beauveria bassiana and other fungi for control of diamondback moth on cabbage seedlings. Journal of Entomological Science, 33, 142-151.
Singh, R., Verma, K. D., Singh, J., Nigam, R., Singh, M. & Kumar, A. (2017). Multiplication of entomopathogenic fungus Beauberia bassiana (Balsamo) on solid and liquid media. Journal of Pharmacognosy and Phytochemistry,
 SP1: 345-351.  
Smith, R. J. & Grula, E. A.  (1981). Nutritional requirments for conidial germination and hyphal growth of Beauveria bassiana. Journal of Invertebrate Pathology, 37,222-230. 
Sujatha, K. Srilakshmi, C. & Lenin, E. A. (2016). Selection of a suitable medium for the mass production of some selected entamopathogenic fungus. International Journal of Recent Scientific Research, 7(7),12370-12372.                                                                
Thomas, K. C., Khachatourians, G. G. & Ingledew, W. M. (1987). Production and properties of Beauveria bassiana conidia cultivated in submerged culture. Journal of Canadian Microbiology, 33, 12- 20.