Production of biopesticides

definition, differences, advantages, production and future approaches

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. R ecommended 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 Methodology

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- 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. Travin , D. Y., Watson, Z. L., Metelev , M., Ward, F. R., Osterman , I. A., Khven , I. M., … Severinov , K. (2019). Structure of ribosome-bound azole-modified peptide phazolicin rationalizes its species-specific mode of bacterial translation inhibition. Nature Communications, 10(1).