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LARGE-SCALE PRODUCTION
OFBIO-CONTROLAGENTS
AND BANKABLE PROJECTS
FOR MASS PRODUCTION
Submitted to:
Dr. Subhash Chander Verma
Dr. Tanuja Banshtu
Submitted by:
Harmanjot Singh
H-2021-04-D
ASSIGNMENT
Bio-control Agent
◦ Biological control agents are living organisms-
parasites, predators, fungi, bacteria and viruses.
◦ Natural enemies of pests, which can intervene the life
cycle of insect pests.
◦ Capable of bringing down the targeted pest
population below economic threshold level (ETL).
◦ Minimize the crop damage.
Major bio agent available for commercial production
The rate of
breeding of
the species
Parasitoids Predator Entomopathogens
Trichogramma chilonis, (egg
parasitoids) - for tomato fruit borer
Cryptolaemus montrouzieri
(Austrtralian ladybird beetle) for
control of several species of mealy
bugs and soft scales
Virus: Nuclear Polyhedrosis Virus
(NPV) for major polyphagous pest
like Helicoverpa armigera and
Spodoptera litura
Trichogramma chilonis for brinjal
shoot and fruit borer, shoot borers of
cotton, sugarcane, rice
Chrysoperla spp. (green lacewing
bug) for the control of aphids, white
flies etc.
Bacteria: Bacillus thuringiences for
control of lepidopterous pests
Namatodes : for control of soil-borne
grubs, lepidopterans and some foliar
pests
Status of Biocontrol agent in India
◦ Sugarcane pyrilla has been successfully controlled in a number of states by the
introduction of its natural enemy Epiricania melanoleuca and Tetrastictus
pyrillae.
◦ Trichogramma, which feeds on the eggs of sugarcane borers, has been used
against the borers in the states of Tamil Nadu, Rajasthan, UP, Bihar, Punjab and
Haryana.
◦ The sugarcane scale insect has been controlled with the help of predatory
coccinellid beetles in UP, West Bengal, Gujarat and Karnataka.
Scope of Large -Scale Production of Biocontrol
Agents
◦ First private Insectary Biocontrol Research laboratory – Bangalore in 1981.
◦ 26 Central Integrated pest management centres in different states.
◦ According to NBAIR, 128 organisation producing bio agents.
 Private companies -80
 Agricultural universities-10
 ICAR Institutes- 8
 Breeding Laboratories-4
 Pest management centre-26
Objectives of Large scale Production of Bio-control
agents
◦ To establish the bankability of mass multiplication of various bio-agents.
◦ To serve as guidelines for extending financial assistance to entrepreneurs
who may be interested in large scale production unit.
◦ To promote setting up of more biocontrol production units.
◦ To disseminate widely the technology.
Bio agents to be used for production of Bioinputs
Group Bioagent Target Insect
1 Trichogramma spp.
(egg parasitoids)
Used for control of borer in
sugarcane, bollworm of cotton,
sorghum, stem borer
Chrysoperla carnea (Chrysopid
predetor)
Controls larval pests in pulses,
vegetables /fruits
Cryptolaemus montrouzieri
(Ladybird beetle)
To control mealy bugs
especially on fruits
Bio Agents to be used for production of Bioinputs
Group Bioagent Target Insects
2 NPV of Helocoverpa armigera and
Spodoptera litura
Used against boll worms in cotton and
pod borers.
Pheromone lures for Helicoverpa
armigera and Spodoptera litura
To trap reproductive males of gram
pod borer and tobacco caterpillar
Bio agents to be used for production
Cryptolaemus montrouzieri
◦ In the laboratory, the life cycle is completed in approximately 30 days.
◦ Both adults and grubs are predating almost all stages of the mealy bug. However the grubs are voracious
feeders. The coccinellid grub consumes a total of 900 to 1500 mealy bug eggs in its development.
◦ After 15 days of infestation of pumpkins with bugs they are exposed to a set of 100 beetles for 24 hrs.
◦ The beetle during the period of exposure feed on mealy bugs as well as deposits their eggs singly or in groups of 4-
12.
◦ The young grubs feed on eggs and small mealy bugs but as they grow they become voracious and feed on all stages of
mealy bugs.
◦ For facilitating the pupation of grubs dried guava leaves or pieces of papers are kept at the base of each of the cages.
◦ The first beetle from the cages starts emerging on 30th day of exposure to C. montrouzieri adults.
◦ The beetles are collected daily and kept in separate cages for about 10-15 days to facilitate completion of mating and
pre-oviposition.
◦ The beetles are also fed on diet containing agar powder (1 g), sugar (20 g), honey (40
ml) and water (100 ml).
◦ The adult diet is prepared by boiling sugar in 70 ml of water, adding 1 g agar,
diluting 40 ml honey in 30 ml of water and adding to the sugar and agar mixture
when it comes to boiling point.
◦ The hot liquid diet is kept on small white plastic cards in the form of droplets which
get solidified on cooling.
◦ Such cards containing adult diet can be fed not only to C. montrouzieri but also to
many other species of coccinellids.
◦ From each cage about 175 beetles are obtained. The emergence of the beetles is
completed within 10 days.
A. Eggs,
B. Grub,
C. Pupa,
D. Adult of Cryptolaemus
montrouzieri
A
B
C
D
Bio agents to be used for production
Trichogramma spp. (egg parasitoids)
◦ The eggs of Corceyra cephalonica treated with with UV rays for 45 minutes to prevent
hatching are glued to ‘Trichocards’ of 15 am × 10 cm.
◦ Apply gum of card and sprinkle the cleaned eggs uniformly in a single layer.
◦ Treat the eggs under UV lamp for 30 mins.
◦ Take the polythene bag, insert UV treated trichocards and nucleus card at 6:1 ratio (6 Corcerya
egg cards: 1 trichogramma card) and place 50 % honey solution in cotton soaked.
◦ Remove the trichocards after 24 hours .
◦ On 4th day, corcerya eggs changes to black in colour and it indicates the parasitization of eggs.
◦ At fourth or 5th day trichocards can be stored in refrigrator at 10⁰C upto 21 days.
Target Insect
Used for control of borer in sugarcane, bollworm of cotton, sorghum, stem borer
◦ Tricho card
Tricho card Trichogramma spp.
Bio agents to be used for production
Chrysoperla carnea (Chrysopid predetor)
◦ In mass production, the adults are fed on various types of diets.
◦ The larvae are either reared in plastic tubes or empty injection vials or in groups in large containers.
◦ The adults are collected daily and transferred to pneumatic glass troughs or G.I. round troughs (30
cm x 12 cm).
◦ Before allowing the adults, the rearing troughs are wrapped inside with brown sheet which act as
egg receiving card.
◦ About 250 adults (60% females) are allowed into each trough and covered with white nylon or
georgette cloth secured by rubber band.
◦ Besides an artificial protein rich diet is provided in semisolid paste form in three spots on the cloth
outside.
◦ This diet consists of one part of yeast, fructose, honey, Proteinex R and water in the ratio 1:1:1:1.
◦ The adults lay eggs on the brown sheet.
◦ The adults are collected daily and allowed into fresh rearing troughs with fresh food. From the old
troughs, the brown paper sheets along with Chrysoperla eggs are removed.
Eggs
Chrysoperla
Nymphs
Storage and Destalking of Eggs
The brown paper sheet kept inside the adult rearing troughs contain large number of eggs each laid on a
stalk or pedicel.
As such the sheets are stored at 10ºC in B.O.D. incubator or refrigerator for about 21 days.
When the eggs are required for culturing or for field release the egg sheets are kept at room temperature for
a day and the eggs during this period turn brown and hatch on 3 days later.
Mass Rearing
◦ It is done in Group Rearing round basins (28 cm dia) @ 250 larvae/basin, covered with khada cloth.
◦ The eggs of Corcyra cephalonica is given as feeding material for the larvae in the laboratory.
◦ For rearing 500 Chrysoperla larvae the total quantity of Corcyra eggs required is 22 cc @ 5 cc/feeding
for five feedings in alternate days.
◦ The Chrysoperla larvae pupate into round white coloured silken cocoons in 10 days.
◦ The cocoons are collected with fine brush and transferred into 1 lit plastic containers with wire mesh
window for emergence of adults.
◦ From the cocoons, pale green coloured adults with transparent lace like wings emerge in 9-10 days.
Bio agents to be used for production
NPV of H. armigera
◦ The NPV of H. armigera is propagated in early fifth instar larvae.
◦ The virus is multiplied in a facility away from the host culture laboratory.
◦ The dose of the inoculum used is 5 x 105 polyhedral occlusion bodies (POB) in 10 ml suspension.
◦ The virus is applied on to the semisynthetic diet (lacking formaldehyde) dispensed previously in 5 ml glass vials.
◦ A blunt end polished glass rod (6 mm) is used to distribute the suspension containing the virus uniformly over the diet
surface.
◦ Early fifth instar stage of larvae are released singly into the glass vials after inoculation and plugged with cotton and
incubated at a constant temperature of 25ºC in a laboratory incubator.
◦ When the larvae exhausted the feed, fresh untreated diet is provided.
◦ The larvae are observed for the development of virosis and the cadavers collected carefully from individual bottles
starting from fifth day.
◦ Approximately, 200 cadavers are collected per sterile cheese cup (300 ml) and the contents are frozen immediately.
◦ Depending upon need, cadavers are removed from the refrigerator and thawed very rapidly by agitation in water.
NPV Infected Larva
Bio agents to be used for production
Pheromone lures
Pheromone: A substance that is secreted by an organism to the outside and causes a specific
reaction in a receiving organism of the same species.
Some of the important female sex pheromones
Considerations for Design of pheromone Traps
S.No. Parameters Ideal Feature(s) Problem(s)
1. Lure Constant attraction Changing release rate, blend
and active space
2. Physical shape Noninhibitory
omnidirectional
Visual and physical
3. Colour Attractive or neutral Nontarget catch (bees)
4. Durability Long lived UV, rain
5. Trapping surface Constant retention rate Glue aging, evaporation
(liquid trapping well)
6. Cost Low cost Manufacturing volume,
complex designs
Basic requirement for establishment of the production
unit
◦ Land
◦ Building and civil works
◦ Plant and Machinery
◦ Raw material
◦ Water
◦ Power
◦ Man power
Man Power
Particulars Group 1
(Predators and Parasitoids)
Group 2
(NPV)
Technical staff 1 3
Skilled labour 2 5
Semi skilled labour 3 10
Total 6 18
Production Unit Size
Group Biocontrol agents Capacity per year
1 Trichgramma cards 30000 cards
Chrysopid grub/larvae 6000
Cryptolaemus beetles 5000 beetles
2 Ha NPV & SINPV Ha NPV-18000 bottles &
SINPV- 12,000 bottles
Pheromone lures 1000 lures
Budget estimate for mass production of Corcyra and
Trichogrammatids
S.No. Particulars Amount
1 Equipments and material (Fixed
Cost)
12,15,000
2 Recurring cost (per annum) 2,10,000
Grand Total 14,25,000
Total Cost of Chrysoperla sp. cost
S.No Particulars Amount for two release in 50ha
(Rs)
1 Equipments and material (Fixed
Cost)
99500
2 Staff and salary 204000
3 Cost of chemicals and other
materials
84500
Total cost (for two release ) 3,88,000
Total Finance of the unit
S.No Particulars Amount (Rs)
1 Total project cost 1,813,000
2 Margin@ 25% 453,250
3 Bank Loan 1517000
Financial analysis
◦ The cash flow statement covering the Benefit Cost Ratio (BCR), Net Present Worth (NPW) and
Internal/financial rate of return (IRR/FRR) have been worked out for the project. Normally the
BCR should be greater than 1, NPW should be positive and IRR should be greater than 15%.
Financial parameters Estimated values
NPW ( in lakhs) 4.90
BCR 1.63
Average DSCR 5.14
Repayment period 7 year inclusive of 1 year grace period
Financial Assistance
◦ The projects on manufacturing biopesticide products would be
considered for refinance support by National Bank. Therefore,
all participating banks may consider financing this activity
subject to their technical feasibility, financial viability and
bankability.
Regulatory measures for quality control
The quality control of commercial bioagents must be strictly enforced by the
Government of India.
The Directorate of Plant Protection Quarantine and Storage, Department of
Agriculture and Cooperation, Ministry of Agriculture, GOI have issued
guidelines/data requirements for registration of bioagents in the country.
THANK YOU

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Large Scale production of natural enemies.pptx

  • 1. LARGE-SCALE PRODUCTION OFBIO-CONTROLAGENTS AND BANKABLE PROJECTS FOR MASS PRODUCTION Submitted to: Dr. Subhash Chander Verma Dr. Tanuja Banshtu Submitted by: Harmanjot Singh H-2021-04-D ASSIGNMENT
  • 2. Bio-control Agent ◦ Biological control agents are living organisms- parasites, predators, fungi, bacteria and viruses. ◦ Natural enemies of pests, which can intervene the life cycle of insect pests. ◦ Capable of bringing down the targeted pest population below economic threshold level (ETL). ◦ Minimize the crop damage.
  • 3. Major bio agent available for commercial production The rate of breeding of the species Parasitoids Predator Entomopathogens Trichogramma chilonis, (egg parasitoids) - for tomato fruit borer Cryptolaemus montrouzieri (Austrtralian ladybird beetle) for control of several species of mealy bugs and soft scales Virus: Nuclear Polyhedrosis Virus (NPV) for major polyphagous pest like Helicoverpa armigera and Spodoptera litura Trichogramma chilonis for brinjal shoot and fruit borer, shoot borers of cotton, sugarcane, rice Chrysoperla spp. (green lacewing bug) for the control of aphids, white flies etc. Bacteria: Bacillus thuringiences for control of lepidopterous pests Namatodes : for control of soil-borne grubs, lepidopterans and some foliar pests
  • 4. Status of Biocontrol agent in India ◦ Sugarcane pyrilla has been successfully controlled in a number of states by the introduction of its natural enemy Epiricania melanoleuca and Tetrastictus pyrillae. ◦ Trichogramma, which feeds on the eggs of sugarcane borers, has been used against the borers in the states of Tamil Nadu, Rajasthan, UP, Bihar, Punjab and Haryana. ◦ The sugarcane scale insect has been controlled with the help of predatory coccinellid beetles in UP, West Bengal, Gujarat and Karnataka.
  • 5. Scope of Large -Scale Production of Biocontrol Agents ◦ First private Insectary Biocontrol Research laboratory – Bangalore in 1981. ◦ 26 Central Integrated pest management centres in different states. ◦ According to NBAIR, 128 organisation producing bio agents.  Private companies -80  Agricultural universities-10  ICAR Institutes- 8  Breeding Laboratories-4  Pest management centre-26
  • 6. Objectives of Large scale Production of Bio-control agents ◦ To establish the bankability of mass multiplication of various bio-agents. ◦ To serve as guidelines for extending financial assistance to entrepreneurs who may be interested in large scale production unit. ◦ To promote setting up of more biocontrol production units. ◦ To disseminate widely the technology.
  • 7. Bio agents to be used for production of Bioinputs Group Bioagent Target Insect 1 Trichogramma spp. (egg parasitoids) Used for control of borer in sugarcane, bollworm of cotton, sorghum, stem borer Chrysoperla carnea (Chrysopid predetor) Controls larval pests in pulses, vegetables /fruits Cryptolaemus montrouzieri (Ladybird beetle) To control mealy bugs especially on fruits
  • 8. Bio Agents to be used for production of Bioinputs Group Bioagent Target Insects 2 NPV of Helocoverpa armigera and Spodoptera litura Used against boll worms in cotton and pod borers. Pheromone lures for Helicoverpa armigera and Spodoptera litura To trap reproductive males of gram pod borer and tobacco caterpillar
  • 9. Bio agents to be used for production Cryptolaemus montrouzieri ◦ In the laboratory, the life cycle is completed in approximately 30 days. ◦ Both adults and grubs are predating almost all stages of the mealy bug. However the grubs are voracious feeders. The coccinellid grub consumes a total of 900 to 1500 mealy bug eggs in its development. ◦ After 15 days of infestation of pumpkins with bugs they are exposed to a set of 100 beetles for 24 hrs. ◦ The beetle during the period of exposure feed on mealy bugs as well as deposits their eggs singly or in groups of 4- 12. ◦ The young grubs feed on eggs and small mealy bugs but as they grow they become voracious and feed on all stages of mealy bugs. ◦ For facilitating the pupation of grubs dried guava leaves or pieces of papers are kept at the base of each of the cages. ◦ The first beetle from the cages starts emerging on 30th day of exposure to C. montrouzieri adults. ◦ The beetles are collected daily and kept in separate cages for about 10-15 days to facilitate completion of mating and pre-oviposition.
  • 10. ◦ The beetles are also fed on diet containing agar powder (1 g), sugar (20 g), honey (40 ml) and water (100 ml). ◦ The adult diet is prepared by boiling sugar in 70 ml of water, adding 1 g agar, diluting 40 ml honey in 30 ml of water and adding to the sugar and agar mixture when it comes to boiling point. ◦ The hot liquid diet is kept on small white plastic cards in the form of droplets which get solidified on cooling. ◦ Such cards containing adult diet can be fed not only to C. montrouzieri but also to many other species of coccinellids. ◦ From each cage about 175 beetles are obtained. The emergence of the beetles is completed within 10 days. A. Eggs, B. Grub, C. Pupa, D. Adult of Cryptolaemus montrouzieri A B C D
  • 11. Bio agents to be used for production Trichogramma spp. (egg parasitoids) ◦ The eggs of Corceyra cephalonica treated with with UV rays for 45 minutes to prevent hatching are glued to ‘Trichocards’ of 15 am × 10 cm. ◦ Apply gum of card and sprinkle the cleaned eggs uniformly in a single layer. ◦ Treat the eggs under UV lamp for 30 mins. ◦ Take the polythene bag, insert UV treated trichocards and nucleus card at 6:1 ratio (6 Corcerya egg cards: 1 trichogramma card) and place 50 % honey solution in cotton soaked. ◦ Remove the trichocards after 24 hours . ◦ On 4th day, corcerya eggs changes to black in colour and it indicates the parasitization of eggs. ◦ At fourth or 5th day trichocards can be stored in refrigrator at 10⁰C upto 21 days.
  • 12. Target Insect Used for control of borer in sugarcane, bollworm of cotton, sorghum, stem borer ◦ Tricho card Tricho card Trichogramma spp.
  • 13. Bio agents to be used for production Chrysoperla carnea (Chrysopid predetor) ◦ In mass production, the adults are fed on various types of diets. ◦ The larvae are either reared in plastic tubes or empty injection vials or in groups in large containers. ◦ The adults are collected daily and transferred to pneumatic glass troughs or G.I. round troughs (30 cm x 12 cm). ◦ Before allowing the adults, the rearing troughs are wrapped inside with brown sheet which act as egg receiving card. ◦ About 250 adults (60% females) are allowed into each trough and covered with white nylon or georgette cloth secured by rubber band. ◦ Besides an artificial protein rich diet is provided in semisolid paste form in three spots on the cloth outside. ◦ This diet consists of one part of yeast, fructose, honey, Proteinex R and water in the ratio 1:1:1:1. ◦ The adults lay eggs on the brown sheet. ◦ The adults are collected daily and allowed into fresh rearing troughs with fresh food. From the old troughs, the brown paper sheets along with Chrysoperla eggs are removed. Eggs Chrysoperla Nymphs
  • 14. Storage and Destalking of Eggs The brown paper sheet kept inside the adult rearing troughs contain large number of eggs each laid on a stalk or pedicel. As such the sheets are stored at 10ºC in B.O.D. incubator or refrigerator for about 21 days. When the eggs are required for culturing or for field release the egg sheets are kept at room temperature for a day and the eggs during this period turn brown and hatch on 3 days later. Mass Rearing ◦ It is done in Group Rearing round basins (28 cm dia) @ 250 larvae/basin, covered with khada cloth. ◦ The eggs of Corcyra cephalonica is given as feeding material for the larvae in the laboratory. ◦ For rearing 500 Chrysoperla larvae the total quantity of Corcyra eggs required is 22 cc @ 5 cc/feeding for five feedings in alternate days. ◦ The Chrysoperla larvae pupate into round white coloured silken cocoons in 10 days. ◦ The cocoons are collected with fine brush and transferred into 1 lit plastic containers with wire mesh window for emergence of adults. ◦ From the cocoons, pale green coloured adults with transparent lace like wings emerge in 9-10 days.
  • 15. Bio agents to be used for production NPV of H. armigera ◦ The NPV of H. armigera is propagated in early fifth instar larvae. ◦ The virus is multiplied in a facility away from the host culture laboratory. ◦ The dose of the inoculum used is 5 x 105 polyhedral occlusion bodies (POB) in 10 ml suspension. ◦ The virus is applied on to the semisynthetic diet (lacking formaldehyde) dispensed previously in 5 ml glass vials. ◦ A blunt end polished glass rod (6 mm) is used to distribute the suspension containing the virus uniformly over the diet surface. ◦ Early fifth instar stage of larvae are released singly into the glass vials after inoculation and plugged with cotton and incubated at a constant temperature of 25ºC in a laboratory incubator. ◦ When the larvae exhausted the feed, fresh untreated diet is provided. ◦ The larvae are observed for the development of virosis and the cadavers collected carefully from individual bottles starting from fifth day. ◦ Approximately, 200 cadavers are collected per sterile cheese cup (300 ml) and the contents are frozen immediately. ◦ Depending upon need, cadavers are removed from the refrigerator and thawed very rapidly by agitation in water.
  • 17. Bio agents to be used for production Pheromone lures Pheromone: A substance that is secreted by an organism to the outside and causes a specific reaction in a receiving organism of the same species. Some of the important female sex pheromones
  • 18. Considerations for Design of pheromone Traps S.No. Parameters Ideal Feature(s) Problem(s) 1. Lure Constant attraction Changing release rate, blend and active space 2. Physical shape Noninhibitory omnidirectional Visual and physical 3. Colour Attractive or neutral Nontarget catch (bees) 4. Durability Long lived UV, rain 5. Trapping surface Constant retention rate Glue aging, evaporation (liquid trapping well) 6. Cost Low cost Manufacturing volume, complex designs
  • 19. Basic requirement for establishment of the production unit ◦ Land ◦ Building and civil works ◦ Plant and Machinery ◦ Raw material ◦ Water ◦ Power ◦ Man power
  • 20. Man Power Particulars Group 1 (Predators and Parasitoids) Group 2 (NPV) Technical staff 1 3 Skilled labour 2 5 Semi skilled labour 3 10 Total 6 18
  • 21. Production Unit Size Group Biocontrol agents Capacity per year 1 Trichgramma cards 30000 cards Chrysopid grub/larvae 6000 Cryptolaemus beetles 5000 beetles 2 Ha NPV & SINPV Ha NPV-18000 bottles & SINPV- 12,000 bottles Pheromone lures 1000 lures
  • 22. Budget estimate for mass production of Corcyra and Trichogrammatids S.No. Particulars Amount 1 Equipments and material (Fixed Cost) 12,15,000 2 Recurring cost (per annum) 2,10,000 Grand Total 14,25,000
  • 23. Total Cost of Chrysoperla sp. cost S.No Particulars Amount for two release in 50ha (Rs) 1 Equipments and material (Fixed Cost) 99500 2 Staff and salary 204000 3 Cost of chemicals and other materials 84500 Total cost (for two release ) 3,88,000
  • 24. Total Finance of the unit S.No Particulars Amount (Rs) 1 Total project cost 1,813,000 2 Margin@ 25% 453,250 3 Bank Loan 1517000
  • 25. Financial analysis ◦ The cash flow statement covering the Benefit Cost Ratio (BCR), Net Present Worth (NPW) and Internal/financial rate of return (IRR/FRR) have been worked out for the project. Normally the BCR should be greater than 1, NPW should be positive and IRR should be greater than 15%. Financial parameters Estimated values NPW ( in lakhs) 4.90 BCR 1.63 Average DSCR 5.14 Repayment period 7 year inclusive of 1 year grace period
  • 26. Financial Assistance ◦ The projects on manufacturing biopesticide products would be considered for refinance support by National Bank. Therefore, all participating banks may consider financing this activity subject to their technical feasibility, financial viability and bankability.
  • 27. Regulatory measures for quality control The quality control of commercial bioagents must be strictly enforced by the Government of India. The Directorate of Plant Protection Quarantine and Storage, Department of Agriculture and Cooperation, Ministry of Agriculture, GOI have issued guidelines/data requirements for registration of bioagents in the country.