This document discusses the production of biopesticides as an alternative to synthetic pesticides. It describes various types of biopesticides including those derived from microorganisms like Bacillus thuringiensis, fungi, viruses and nematodes. The document outlines the manufacturing process for biopesticides including using solid-state fermentation to produce fungal conidia. It also discusses some commonly used biopesticides like pyrethrum, neem and pheromones. The benefits of biopesticides over chemical pesticides are noted and future research opportunities around nano-enabled biopesticide formulations are proposed.

1. PRODUCTION OF BIOPESTICIDES
 SUSHMITA DHIR
(19MBT0009)

2. INTRODUCTION
• Pesticides that are naturally
produced are called biopesticides
and have been attracting interest
because they are an alternative to
synthetic pesticides for the
protection of plant crops.
• Recommended as potentially good
alternatives to synthetic pesticides
• Biopesticides may be derived from
animals (e.g. nematodes), plants
(Chrysanthemum, Neem) and
microorganisms (e.g. Bacillus
thuringiensis, Trichoderma,
nucleopolyhedrosis virus).
Fig 01 : Consumption of biopesticides
Fig 02 : Biopesticides Registered under
Insecticides Act, 1968
Source : Vachon, V., Laprade, R., & Schwartz, J.-L. (2012).
Current models of the mode of action of Bacillus
thuringiensis insecticidal crystal proteins: A critical
review. Journal of Invertebrate Pathology, 111(1), 1–12.

3. Table 01 : Biopesticide v/s Synthetic pesticide
BIOPESTICIDE SYNTHETIC PESTICIDE
• Typically designed to affect only the
target pest or groups of specific
organisms
• inherently less toxic than conventional
pesticides.
• Biopesticides often are effective in
very small quantities and often
decompose quickly, resulting in lower
exposures and largely avoiding the
pollution problems caused by
conventional pesticides.
• Biodegradability (natural or derived
from living organisms or their
metabolites)
• biopesticides can greatly reduce the
use of conventional pesticides, while
crop yields remain high.
• Do not exhibit specificity in their performance
hence, present toxicity to the pests and pathogens
contaminants of plant crops
• The increased exposure of humans to these
substances, may cause some diseases, including
Parkinson’s disease, type 2 diabetes, certain types
of cancers, endocrine disruption, neurotoxicity and
even obesity
• accumulate in the human body
• Pesticide residues can leach the subsoil and
contaminate groundwater
• continuous use of synthetic pesticides makes them
more resistant pests
• synthetic pesticides such as : organochlorines,
organophosphates, carbamates and
organophthaloids

4. Table 02 : Classifications of Biopesticides
Biochemical pesticides Microbial pesticides Plant-Incorporated-
Protectants (PIPs)
• Biochemical pesticides are
naturally occurring
substances that control pests
by non-toxic mechanisms.
• Biochemical pesticides
include substances that
interfere with mating, such
as insect sex pheromones, as
well as various scented plant
extracts that attract insect
pests to traps.
• Microbial pesticides consist
of a microorganism (e.g., a
bacterium, fungus, virus or
protozoan) as the active
ingredient.
• Microbial pesticides can
control many different kinds
of pests, although each
separate active ingredient is
relatively specific for its
target pest[s]. For example,
there are fungi that control
certain weeds and other
fungi that kill specific
insects.
• The most widely used
microbial pesticides are
subspecies and strains of
Bacillus thuringiensis, or Bt.
• Plant-Incorporated-
Protectants (PIPs) are
pesticidal substances that
plants produce from genetic
material that has been added
to the plant.
• For example, scientists can
take the gene for the Bt
pesticidal protein and
introduce the gene into the
plant's own genetic material.
• Then the plant, instead of
the Bt bacterium,
manufactures the substance
that destroys the pest. The
protein and its genetic
material, but not the plant
itself.

5. SOME OF THE IMPORTANT MICROBIAL
PESTICIDES
 Bacillus thuringiensis
• Spores and crystalline insecticidal
proteins of B. thuringiensis used to
control insect pests
• Applied as liquid sprays
• Highly specific,
environmentally friendly, with
little or no effect on humans,
wildlife, pollinators, and most
other beneficial insects, and are
used in organic farming;
• Control lepidopterous pests like
american bollworm in cotton
and stem borers in rice.
• When ingested by pest larvae,
Bt releases toxins which damage
the mid gut of the pest,
eventually killing it.
Fig 03 : Mode of action of Bacillus thuringiensis in pest control
Source : Rodríguez, P., Cerda, A., Font, X., Sánchez, A., & Artola, A. (2019).
Valorisation of biowaste digestate through solid state fermentation to produce
biopesticides from Bacillus thuringiensis. Waste Management, 93, 63–71.

6. AGROBACTERIUM RADIOBACTER
(AGROCIN)
• Agrobacterium radiobacter is
used to treat roots during
transplanting, that checks
crown gall.
• Crown gall is a disease in
peaches, grapevine, roses and
various plants caused by soil
borne pathogen Agrobacterium
tumefaciens.
• The effective strains of A.
radiobacter posses two
important features: They are
able to colonize host roots to a
higher population density.
They produce an antibiotic,
agrocin, that is toxic to A.
tumefaciens.
Fig 04 : Mode of action of Agrobacterium in a plant cell
Source : Hwang, H. H., Yu, M., & Lai, E. M. (2017). Agrobacterium-
mediated plant transformation: biology and applications. The arabidopsis
book, 15, e0186.

7. PLANT BIOPESTICIDES
• Plants that produce substances or
chemicals that have detrimental effect
on the pest organism
• Pyrethrum (Chrysanthemum) flowers
contain active pyrethrins extracted and
sold in the form of an oleoresin. This is
applied as a suspension in water or oil, or
as a powder. Pyrethrins attack the nervous
systems of all insects, and inhibit female
mosquitoes from biting and insect
repelling.
• Neem does not directly kill insects on the
crop. It acts as an anti-feedant, repellent,
and egg-laying deterrent, protecting the
crop from damage. The insects starve and
die within a few days. Neem also
suppresses the hatching of pest insects
from their eggs.
Fig 05 : Pyrethrum (Chrysanthemum)
Fig 06 : Neem

8. BIOCHEMICAL PESTICIDES
• They are naturally occurring substance
to control pest by non-toxic
mechanisms.
• Biochemical pesticides include
substances as insect sex pheromones,
that interfere with mating that attract
insect pest to traps.
• The synthetic attractants are used in
one of four ways:
• As a lure in traps used to monitor pest
populations
• As a lure in traps designes to trap out a
pest population
• As a broadcast signal intended to
disrupt insect mating
• As an attractant in a bait containing an
insecticide
Fig 07 : Rice Weevil (Sitophilus oryzae)
pheromone tra

9. PLANT-INCORPORATED-PROTECTANTS
(PIPS)
• Plant-incorporated
protectants are pesticidal
substances produced by plants
and the genetic material
necessary for the plant to
produce the substance
• For example, scientists can
take the gene for a specific Bt
pesticidal protein and
introduce the gene into the
plant's genetic material
• The new Bt cotton product
contains the dual genes Cry
IA(c) and Cry IF, transformed
with Agrobacterium
tumefaciens and incorporated
through back crossing Source : Wang, Y., Wang, J., Fu, X., (2019). Bacillus thuringiensis Cry1Da_7 and
Cry1B.868 Protein Interactions with Novel Receptors Allow Control of Resistant Fall
Armyworms, Spodoptera frugiperda (J.E. Smith). Applied and environmental
microbiology, 85(16), e00579-19.
Fig 08 : plant-incorporated-protectants action

10. MANUFACTURING PROCESS
Flow chart 01 : Schematic representation of Biopesticide manufacturing process

11.  RAW MATERIAL
•May be organic or inorganic compounds
•Different raw material for different pesticide
 REACTOR SYSTEM
•Chemical process takes place in the presence of chemicals such as oxidation, nitration, condensation, etc.
 FRACTIONATION SYSTEM
•Separation process in which certain quantity of a mixture (solid, liquid, solute, suspension or isotope) is
divided up in a number of smaller fractions in which composition change
•Recovery
 DRYER
•Removal of water or other solvent by evaporation from solid, semi-solid or liquid
•Final production step before selling or packaging products.
 SCRUBBERS
•To remove priority pollutants from pesticide product using scrubbing liquor
•Wastewater go to treatment plant
 PACKAGING
•Packed in dry and clean containers e.g., drums type depend on type of pesticide
•Capacity 10,25,50,100,200 litres.
•Temper-proof, closer to avoid leakage, sturdy
 FORMULATION
•Processing a pesticide into granules, liquid, dust and powder to improve its properties of storage,
handling, application, effectiveness, or safety.
•Dry mixing, grinding of solids, dissolving solids and blending

12. PRODUCTION OF CONIDIA BY THE FUNGUS METARHIZIUM
ANISOPLIAE USING SOLID-STATE FERMENTATION
• Solid-state fermentation (SSF)
is the preferred system to
produce conidia from
entomopathogenic fungi
mainly using trays of plastic
bags containing substrates
such as rice or other solid
agricultural wastes which
sometimes are supplemented
or combined in order to
achieve higher conidial yields
• Conidia, are related to
virulence against insect
• Conidia production of M.
Anisopliae under two different
techniques using SSF: plastic
bags and tubular bioreactors
Fig 09 : Solid-state fermentation and respirometric analysis
apparatus. ( a ) Air distributor, ( b ) Water bath, ( c ) Solid- state
culture bioreactors , ( d ) Air dryers, ( e ) Respirometer for
CO 2 , O 2 , and air fl ow rate measure and ( f ) Computer
Source : Loera-Corral, O., Porcayo-Loza, J., Montesinos-Matias, R., & Favela-Torres, E. (2016). Production of Conidia by the Fungus
Metarhizium anisopliae Using Solid-State Fermentation. Microbial-Based Biopesticides, 61–69.

13. Fig 10 : Common reactors designs in SSC of entomopathogenic fungi with some variables
affecting conidial yields and quality which are also susceptible for optimisation
Source : Muñiz-Paredes, F., Miranda-Hernández, F., & Loera, O. (2017). Production of conidia by
entomopathogenic fungi: from inoculants to final quality tests. World Journal of Microbiology and
Biotechnology

14. Materials required
4.Conidia production in
tubular bioreactor
3. Conidia production in
plastic bag
2.Culture media
1.Organisms
Methodology
5. Evaluation of conidia quality
4. Conidia production in plastic bags and
tubular bioreactors
3. Inocula production
2. Strain conservation
1. Metarhizium Anisopliae propagation and
M.Anisopliae reactivation in tenebrio molitor

15. CONCLUSION
• An ecofriendly alternative to chemical pesticides is biopesticides, which encompasses
a broad array of microbial pesticides, biochemicals derived from micro-organisms and
other natural sources, and processes involving the genetic incorporation of DNA into
agricultural commodities that confer protection against pest damage
• Bacillus species are well known producers of antimicrobial compounds and as such
are of great interest in the fight against plant pathogens
• The manipulation of culture conditions in SSC leading to optimal conidial yields
could affect the quality required for outstanding abiotic factors, such as those found
after application in crop fields. In this context, some promising areas for research are
those related with the quality of the inoculants and the inclusion of sub-lethal stress
conditions to generate cross-protection, which also should be considered in the design
of improved bioreactors. The knowledge and advances achieved in these optimisation
procedures are relevant for better products in the strong market of mycopesticides
• The SSF system is useful for spores production of BCA’s microorganisms used as
biopesticides. Also, SSF facilitates development of formulations used in field crops,
will decrease process costs. Production costs of biopesticides by SSF are low because
of the use of natural substrates (mainly by-products), low aeration rate and bioreactors
that can be used once.

16. FUTURE RESEARCH
• In the present context of climate change, Bt is
the most promising biopesticide because it is
relatively more effective at high temperatures as
well as having extended shelf-life during storage
• Environmental safety concerns have resulted in
increased demand for Bt-based pesticides and
formulations
• Certain drawbacks that exist in conventional Bt
biopesticides have led to a search for newer
approaches to improve their efficacy
• In this new era, Bt in combination with
nanoscience in crop protection is an unexplored
area. Therefore, thrust should be given to the
development of nano-Bt formulations with
higher efficacy, efficient delivery, reduction in
dosage rate, a faster mode of action, and
increased field persistence
• Nanotechnology holds promise for further
improving the efficacy of Bt through particle
size reduction as well as delivery of Cry toxins.
Fig 09 : Application of nanotechnology in pesticide
delivery
Source : Mishra, S., Keswani, C., Abhilash, P. C., Fraceto, L.
F., & Singh, H. B. (2017). Integrated Approach of Agri-
nanotechnology: Challenges and Future Trends. Frontiers in
plant science, 8, 471.

17. REFERENCES
• Glare, T. R., Gwynn, R. L., & Moran-Diez, M. E. (2016). Development of
Biopesticides and Future Opportunities. Microbial-Based Biopesticides, 211–221.
• Shapiro-Ilan, D. I., Morales-Ramos, J. A., & Rojas, M. G. (2016). In Vivo
Production of Entomopathogenic Nematodes. Microbial-Based Biopesticides, 137–
158.
• Morán-Diez, M. E., & Glare, T. R. (2016). What are Microbial-based
Biopesticides? Microbial-Based Biopesticides, 1–10.
• Muñiz-Paredes, F., Miranda-Hernández, F., & Loera, O. (2017). Production of
conidia by entomopathogenic fungi: from inoculants to final quality tests. World
Journal of Microbiology and Biotechnology
• Loera-Corral, O., Porcayo-Loza, J., Montesinos-Matias, R., & Favela-Torres, E.
(2016). Production of Conidia by the Fungus Metarhizium anisopliae Using Solid-
State Fermentation. Microbial-Based Biopesticides, 61–69. doi:10.1007/978-1-
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Journal of Invertebrate Pathology, 111(1), 1–12.
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