Exercise 3. Handling and Mass Production of Biological Control Agents

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MODULE 3
HANDLING AND MASS PRODUCTION OF BIOLOGICAL CONTROL
AGENTS
INTRODUCTION
The production of biological control agents may involve in vivo or in vitro
methods (solid or liquid fermentation). For a particular BCA such as
entomopathogenic nematodes or EPN, the simplest and cheap technology for
production is in vivo method, that involves the use of living insects, mostly larvae
of the greater wax moth (Galleria mellonella), rice moth (Corcyra cephalonica) or
mealworms (Tenebrio molitor) that are very susceptible to EPN infection, and their
bodies contain enough nutrients for EPN reproduction. However, this method is
labor and cost-effective only at a small scale, and it is therefore appropriate for
laboratory use or small-scale applications (Askary and Ahmad, 2017; Puza et al.,
2016; Shairo-Ilan et al., 2012). For fungal BCAs such as hyperparasites and egg
parasites, in vitro culture, which includes both the solid and liquid culture methods,
can be used to culture them monoxenically. It will also ensure consistency in quality
and achieve predictability of biopesticides. To culture them in a more subsistence
or small-scale production, many use organic substrates to mass multiply the
organisms.
OBJECTIVES
1. To determine the growth and development of BCA in organic

substrates.
2. To determine the substrates that will produce the most harvest for
BCA.
MATERIALS
Rice bran Disinfectant (Clorox, ethyl alcohol)

Corn grits Stove
Mushroom compost Autoclave
Saw dust Pentel Pen
Alcohol lamp Weighing scale
Inoculating loop Cotton
Match Isolation box
Pure culture of antagonist Incubation box
Erlenmeyer flask-250 ml
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METHODS
a. Preparation of substrate – weigh 200 g each of the substrate that will be used
and place them in a 250 ml Erlenmeyer flask.
b. of substrate – sterilize the substrate in an autoclave at 15 psi for 1 hour. Allow
the substrate to cool before inoculating the fungal antagonist.
c. Inoculation – inoculate a loopful of the antagonist in the sterilized substrate
following the aseptic procedure as you perform the inoculation process. In
the absence of laminar flow, an improvised isolation/inoculation chamber can
be used. This is to minimize possible contamination.
d. Incubation – after the inoculation process, incubate the set-up in a chamber
with light to allow the antagonist to grow. Start observing for growth of the
pathogen after three days and continue observing until the flask is filled with
spores and mycelia.
WATCH ME! TRICHODERMA SP. MASS PRODUCTION ON VAR IOUS
SUBSTRATES
https://drive.google.com/file/d/1Lh77OCMpjyi3sWmlD54WQaP79px22n1M/vi
ew?usp=sharing
WORKSHEET 3. HANDLING AND MASS PRODUCTION OF BIOLOGICAL CONTROL

AGENTS
ILLUSTRATION
Read and watch the methodology on how to mass produce biological control agent
Trichoderma sp. and draw an illustration of the step-by-step process of mass producing
it.
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This part has been modified from Module 3. Instead of you performing the experiment,
I’ve prepared my own set-up using available substrates. Write your observations and answer the
questions that follow. You may also create your own tables.
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OBSERVATIONS
a. Observe any growth of the antagonist, indicate by filling the space
with + or – sign
Substrates Week after incubation

3 DAI 7 DAI
Corn grits
Rice Bean
Weed
Johnson grass
LEARNING ASSESSMENT: Answer the following questions
1. Which of the substrate was first colonized by the antagonist?
2. Describe the growth of the antagonist in the different substrates

after three days? After seven days?
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3. Based on your observation, which of the four substrates is best to

grow the antagonist? Why?
WORKSHEET 3.1. RESEARCH ACTIVITY
Find one research article on the production of Biological Control Agents on

various substrates and write your own summary of which substrate worked best and
what factors drove the Biological Control Agent's growth. Write a single-paragraph
summary of the article consisting of 300 words minimum.
*Cite your reference

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1

1. To determine the growth and development of BCA in organic

Rice bran Disinfectant (Clorox, ethyl alcohol)

WORKSHEET 3. HANDLING AND MASS PRODUCTION OF BIOLOGICAL CONTROL

Substrates Week after incubation

LEARNING ASSESSMENT: Answer the following questions

1. Which of the substrate was first colonized by the antagonist?

2. Describe the growth of the antagonist in the different substrates

3. Based on your observation, which of the four substrates is best to

WORKSHEET 3.1. RESEARCH ACTIVITY

Find one research article on the production of Biological Control Agents on

*Cite your reference

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