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Inhibition of Metarhizium anisopliae infection of Protaetia brevitarsis seluensis larvae using several effective microorganisms

  • Kwak, Kyu-Won (National Institute of Agricultural Science, Department of Agricultural Biology) ;
  • Kwon, Soon Woo (National Institute of Agricultural Science, Department of Agricultural Biology) ;
  • Nam, Sung-Hee (National Institute of Agricultural Science, Department of Agricultural Biology) ;
  • Park, Kwan-Ho (National Institute of Agricultural Science, Department of Agricultural Biology) ;
  • Kim, Eun-Sun (National Institute of Agricultural Science, Department of Agricultural Biology) ;
  • Lee, Hee-Sam (National Institute of Agricultural Science, Department of Agricultural Biology) ;
  • Choi, Ji-Young (National Institute of Agricultural Science, Department of Agricultural Biology) ;
  • Han, Myung-Sae (Kyungpook National University, Department of Bio-fibers and Materials Science)
  • Received : 2017.09.04
  • Accepted : 2018.02.01
  • Published : 2018.03.31

Abstract

The purpose of this study was to determine the best method for minimizing the occurrence of Metarhizium anisopliae infection of Protaetia brevitarsis seluensis during mass breeding on agricultural farms. There is a high demand for the use of P. b. seluensis larvae in animal feed and as food for humans. However, mass breeding results in the entomopathogenic fungal (usually M. anisopliae) infection of P. b. seluensis. A mixture of microorganisms (Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae) delayed fungal infection by M. anisopliae, which infected fewer P. b. seluensis when the microorganism mixture was added to sawdust as feed for P. b. seluensis. When sawdust with the effective microorganisms (EM) was given to P. b. seluensis for 30 d, their mortality rate was approximately 35 % less than that of the control group, which was fed sawdust without the EM. In addition, the growth of M. anisopliae on agar media spread with each bacterium as inhibited by up to 80 % more than those spread with 4 % sodium hypochlorite, which is a harmless fungal inhibitor generally used in agricultural farms for disinfection.

Keywords

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Fig. 1. Metarhizium anisopliae infection of Protaetia brevitarsissleuensis at a farm in Korea.P. b. seluensis that died from M. anisopliae infection at P. b. seulensisfarms in Wanju and Cheongju, Korea, show dark green fungicovering their whole bodies.

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Fig. 2. Comparison of Protaetia brevitarsis seluensis mortality betweenEM treatment and control groups in Metarhizium anisopliae sawdust.The mortality rate of the third instar P. b. seluensis larvaeunder both the sealed and ventiliated conditions was 100%after 30 days, but the mortality rate of the EM group was 23.3%.(Con. Ven.: Control ventilation condition, Con. Seal.: Control sealedcondition, A. seal.: Effective microorganism composite, sealed condition)(t-test: *** P < 0.001)

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Fig. 3. Measurement of the growth inhibition zone and rate (%)against Metarhizium anisopliae for each EM. (A) In Sabourauddextrose agar medium, 200 μL of triple distilled water as a negativecontrol and 4 % sodium hypochlorite as a positive control wereevenly smeared using a sterilized glass rod before seeding M.anisopliae. A sample of 200 μL of Lactobacillus plantarum, Bacillussubtilis, and Saccharomyces cerevisiae were spread onto individual

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Fig. 4. Measurement of mortality rate when EM was injected intoProtaetia brevitarsis seluensis by hemolymphatic injection.Mortality rate was examined when each EM was injected into third

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Fig. 5. SEM of Metarhizium anisopliae treated the specificconcentrations of lactic acid The morphology of M. anisopliaeon SEM appeared not to be conidia but rather mycelia, andfungal sexual reproduction decreased. The size of sporesincreased and the mycelium became denser when treated lacticacid concentration to M. anisopliae was lower. This indicatedfungal conidial production was affected by specifc concentrationof lactic acid.

Acknowledgement

Supported by : National Institute of Agricultural Science

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