1. Plant Pathology Research Institute, AARI, Faisalabad

2. White fly control by the use of entomopathogenic fungi
Muhammad Nasir*, Mumtaz Hussain*, Babar Iqbal*, Munir A. Shehzad * & Shaukat Ali **
* Plant Pathology Research Institute, AARI, Faisalabad,
** South China Agriculture University, Guangzhou, China
Corresponding
Plant Pathology Research Institute, AARI, Faisalabad

3. Whitefly More than 1200 species
Most damaging 1 to crop plants is Bemisia tabaci
Mostly found in tropical, subtropical & temperate zones
Infested many crops & was not a major pest prior to the mid 1980s
Since 1992, CLCuV disease transmitted by B. tabaci has decreased cotton yield significantly.
Being vector of CLCuV, all cotton varieties are heavily affected till to date
Estimated losses more over half billion dollars
due to this loss, growers forced to produce alternate crops
Insecticides give good control but not sustainable, many disadvantages)
Biological control (entomopathogens) – alternative approach; many advantages: e.g. envt. friendly, safe to NEs and users
Ex. of entomopathogens: Isaria, Beauveria, Lechanicillium etc.
Plant Pathology Research Institute, AARI, Faisalabad

4. Whitefly damage 3 types of damages caused by Bemisia tabaci:
Direct damage: (piercing & sucking sap from plant foliage)
Indirect damage: ( honey dew, sooty mold)
Virus Transmission: (transmit 111 viral diseases in different crops)
Plant Pathology Research Institute, AARI, Faisalabad

5. Ecological control of crop diseases & pests
Traditional agriculture Industrial agriculture
Industrialization
Global warming
Developed & developing Countries i.e. France, America, Australia, China
Disruption
maintain
Organic Agriculture
Natural Ecosystem
Ecological control of crop diseases & pests
Biological Control
Entomopathogenic fungi
Plant Pathology Research Institute, AARI, Faisalabad

6. Back ground
Plant Pathology Research Institute, AARI, Faisalabad

7. Many PESTS attack to crops…
Plant Pathology Research Institute, AARI, Faisalabad

8. Scientists invented or formulated different kinds of PESTICIDES…
Plant Pathology Research Institute, AARI, Faisalabad

9. and they were very effective…
Plant Pathology Research Institute, AARI, Faisalabad

10. Pesticides applied in different crops…
Plant Pathology Research Institute, AARI, Faisalabad

11. 1. Natural enemies (friendly insects) are
affected much by the pesticides…
Plant Pathology Research Institute, AARI, Faisalabad

12. 2. Toxic pesticide residues on the crop
that could even result to…
Plant Pathology Research Institute, AARI, Faisalabad

13. High risk for the health of the farmers, their families and animals…
Plant Pathology Research Institute, AARI, Faisalabad

14. 4. Non-target organisms including birds are also affected…
Plant Pathology Research Institute, AARI, Faisalabad

15. 5. Environmental pollution…
Plant Pathology Research Institute, AARI, Faisalabad

16. 6. Development of pests RESISTANCE…
Minor pests can become more serious than key pests
Plant Pathology Research Institute, AARI, Faisalabad

17. The “3R” problems of synthetic chemical pesticide (Resurgence of pest insects, Resistance of insects to insecticide, Residue of insecticide) .
Resurgence
Public health
Residue
Resistance
Plant Pathology Research Institute, AARI, Faisalabad

18. Effect of pesticides to the pests is lesser compared to its
toxic effect to the env’t., and non-target organisms…
Plant Pathology Research Institute, AARI, Faisalabad

19. “GLOBAL WARMING” = climate change…
Pesticides may have contributed a fraction
to the problem of…
“GLOBAL WARMING”
= climate change…
Plant Pathology Research Institute, AARI, Faisalabad

20. Look for other alternative approaches…
Plant Pathology Research Institute, AARI, Faisalabad

21. biological control agents…
Scientists & experts studied the use of
biological control agents…
Plant Pathology Research Institute, AARI, Faisalabad

22. agents… Taking care of the biological control
Plant Pathology Research Institute, AARI, Faisalabad

23. Biological control method is now
on the go…
Plant Pathology Research Institute, AARI, Faisalabad

24. Biological control
Biological control is the use of a living organism for pest management.
In the past 5 years bio control agents have increased from 1% to 3%.
Annual growth rate is more than10%.
Plant Pathology Research Institute, AARI, Faisalabad

25. Biological control Classical biological control can be achieved by
Introduction of natural enemies from their native range into another part or country to control an exotic pest
Conservation of natural enemies
Mass rearing and release of natural enemies &
Production of biological pesticides – in broad sense biological control
Plant Pathology Research Institute, AARI, Faisalabad

26. Biological Control Advantages:
Safe to the natural enemies (friendly or beneficial insects)
2. Cheaper than synthetic pesticides
3. Non-toxic to the farmers, farm produce, and to
other living organisms
4. Does not pollute to the environment
5. Sustainable
6. No development of insect pest resistance
7. Maintain the ecological balance of nature
Plant Pathology Research Institute, AARI, Faisalabad

27. Biological control agents
Parasites / parasitoids
Predators
Entomopathogens
Plant Pathology Research Institute, AARI, Faisalabad

28. Entomopathogens
Entomopathogenic microbes are the most promising bio agents in the next decades. They are not only safe and cheap, & also are so fast in killing the targets.
Entomopathogenic fungi:
Insect pathogenic bacteria
Insect pathogenic nematodes
Insect pathogenic viruses
Plant Pathology Research Institute, AARI, Faisalabad

29. Entomopathogenic Fungi
Fungal diseases among insects are common and all insect orders are susceptible to fungal diseases.
About 750 species of entomopathogenic fungi are currently known.
10 species are being used or developed for insect control and the full potential of these fungi has not been approached.
Plant Pathology Research Institute, AARI, Faisalabad

30. Natural occurrence of entomopathogenic fungi
Entomopathogenic fungi are distributed in a wide range of habitats including
aquatic
forest,
agricultural land,
pasture,
desert, and
urban habitats
Soil is considered an excellent environmental shelter for entomopathogenic fungi
Infected insect pests
Plant Pathology Research Institute, AARI, Faisalabad

31. Natural occurrence of entomopathogenic fungi
Feeding pests:
Locusts, hairy caterpillars, Plutella spp.,
Locust/Metarhizium silkworm/Beauveria moth/Isaria
Plant Pathology Research Institute, AARI, Faisalabad

32. Natural occurrence of entomopathogenic fungi
Fruit or stem bores of crops
Helicoverpa, Spodoptera spp., Beetle grub, white grub / root grubs
Plant Pathology Research Institute, AARI, Faisalabad

33. Natural occurrence of entomopathogenic fungi
Sucking pests:
whitefly, aphid, stinkbug, Spider mites, scale insect
whitefly/nymph stinkbug cicada
Whitefly/adult Spider mite Citrus whitefly
Plant Pathology Research Institute, AARI, Faisalabad

34. Entomopathogenic Fungi
Entomopathogenic fungi are unique compared to other microbes causing diseases in insects because
They infect through the insect cuticle
Do not require to be ingested
Most reports of natural fungal infections of pest refer to
Metarhizium
Beauveria
Isaria
Paecilomyces
Lecanicillium
Aschersonia
Acremonium
Cladosporium
Aspergillus
Zoophthora
Fusarium
Conidiobolus
Plant Pathology Research Institute, AARI, Faisalabad

35. Entomopathogenic Fungi
Most of these fungi have proven to develop for microbial control applications, such as,
Beauveria bassiana
Metarhizium anisopliae
Metarhizium flavoviride
Lecanicillium muscarium
Aschersonia aleyrodis
Isaria fumosorosea
Isaria javanicus
Isaria farinosus
Paecilomyces lilacinus
Conidiobolus coronatus
Zoophthora radicans
Plant Pathology Research Institute, AARI, Faisalabad

36. Whitefly Pathogens:-
Owing to feeding mechanisms of whiteflies, only pathogenic microbes with the ability to penetrate the cuticle (e.g., fungi) have potential as microbial control agents.
More than 20 species of entomopathogenic fungi are known to infect whiteflies
Plant Pathology Research Institute, AARI, Faisalabad

37. Natural occurrence of entomopathogenic fungi on Bemisia populations
Fungus
Location
Source
Hyphomycetes
Aschersonia aleyrodis
Taiwan
Yen and Tsai (1969)
Aschersonia andropogonis
USA
Berger (1921)
Aschersonia cf. goldiana
Brazil
Lourencao et al. (1999)
Beauveria bassiana
Israel
Ben-Ze’ev et al. (1994)
Paecilomyces farinosus
Greece
India
Kirk et al. (1993)
Nene (1973)
Paecilomyces fumosoroseus
(Isaria fumosorosia)
Venezuela
Mexico
Cuba
Trinidad
Hawaii
Nepal
Pakistan
Indonesia
Philippines
Japan
Sosa-Gomez et al. (1997)
R. Hall (pers. communication)
Garza Gonzalez (1993)
Castineiras (1995)
Hall et al. (1994)
R. Humber (pers. communication)
Carruthers et al. (1993)
Balakrishnan and Nene (1980), Lacey et al. (1993)
Lacey et al. (1993)
Humber (1992)
T. Poprawski and R. Carruthers (pers. communication)
S. Kurogi (pers. communication)
Plant Pathology Research Institute, AARI, Faisalabad

38. Natural occurrence of entomopathogenic fungi on Bemisia populations
Fungus
Location
Source
Verticillium lecanii
(Lecanicillium muscarium)
Colombia
Venezuela
Mexico
Israel
Denmark
Spain
Japan
Drummond et al. (1987)
R. Hall (pers. communication)
Nier et al. (1991)
Ben-Ze’ev (1993)
R. Humber (pers. communication)
Lacey et al. (1993)
S. Kurogi (pers. communication)
Entomophthorales
Conidiobolus spp.
USA
Ben-Ze’ev (1993), Gindin and Ben-Ze’ev (1994)
R. Carruthers (pers.communication)
Entomophthora sp.
Zoophthora (Erynia) radicans
Chad
Silvie and Papierok (1991)
Ben-Ze’ev et al. (1988)
Unidentified species
Brazil
Sosa-G! omez et al. (1997)
S. Wraight (unpublished observation)
Plant Pathology Research Institute, AARI, Faisalabad

39. General Fungal infection Process
Plant Pathology Research Institute, AARI, Faisalabad

40. Methodology adopted
Isaria fumosorosea isolation was carried out from whitefly nymph on Sabouraud Dextrose Agar medium & purified & multiplied on PDA & incubated for 10 days at 26 0c
Conidia suspension: 1×103 to 1×107 conidia/ml
Before suspension preparation, viability is determined on PDA
Sprayed the prepared suspension
in cages within laboratory on whitefly nymphs & adults
Mortalities of B. tabaci was recorded after 6 &12 days of treatment
Infective mortalities, the cadavers was taken out and separately cultured at 260C and relative humidity >95% to encourage sporulation. Sporulation of Isaria fumosorosea was observed on cadavers, and considered as a result of infection by Isaria fumosorosea
Infected Whitefly adults & nymphs
Isaria fumosorosea
Plant Pathology Research Institute, AARI, Faisalabad

41. Most commonly observed fungal pathogens
Whitefly Pathogens:-
Most commonly observed fungal pathogens
Fungus
Whitefly infection
Isaria fumosorosea (Wize) Smith
All stages are effected by this fungus
Aschersonia aleyrodis Webber:
Only kills pupae and nymph
Lecanicillium muscarium (Zimmerman) Viegas
Mortality causes up to 96%
Beauveria bassiana (Balsamo) Vuillemin
Only kills Nymph & adults (75-80%)
Metarhizium anisopliae (Metsch.) Sorokin
causes mortality more than 75%
Conidiobolus coronatus
infecting only B. tabaci adults &
causes 95% mortality
Plant Pathology Research Institute, AARI, Faisalabad

42. Entomopathogenic Fungi
Isaria fumosorosea
Most important natural enemy of whiteflies throughout the worldwide
causes the sickness called “Yellow Muscardine”
Strong epizootic potential against Bemisia and Trialeurodes spp. in both greenhouse and open field environments.
grow extensively over the leaf surface under humid conditions is its characteristic that enhances its ability to spread rapidly through whitefly populations.
All stages of white flies are effected by this fungus.
causes mortality levels ranging from 68% to 94% against whitefly
This fungus has been used as a mycopesticide for whitefly management in the United States, Europe, China etc. both in greenhouse as well as in open environment.
Plant Pathology Research Institute, AARI, Faisalabad

43. Entomopathogenic Fungi
Bemisia tabaci adults infected with Isaria
Nymph of infected with
Isaria fumosorosea
Plant Pathology Research Institute, AARI, Faisalabad

44. Entomopathogenic Fungi
Isaria fumosorosea
Isaria javanicus
Isaria farinosus
Paecilomyces lilacinus
Plant Pathology Research Institute, AARI, Faisalabad

45. Entomopathogenic Fungi
Lecanicillium muscarium
Major microbial biocontrol agent of
whiteflies and
aphids.
causes whitefly mortality from 89 to 96 %.
Commercialized worldwide as the biopesticides
Mycotal against whiteflies and thrips and as
Verticillin against whiteflies, aphids and mites.
Fungal growth on infected insect pest
Plant Pathology Research Institute, AARI, Faisalabad

46. Entomopathogenic Fungi
Aschersonia aleyrodis
Only kills pupae and nymph of whitefly
Causes whitefly mortality more than75%
fungus has been developed commercially by Koppert Biological Systems, Holland as a biopesticide
Cotton whitefly
citrus whitefly parasitized by Aschersonia aleyrodis
Nymph of Bemisia tabaci infested with
Aschersonia aleyrodis
Plant Pathology Research Institute, AARI, Faisalabad

47. Entomopathogenic Fungi
Beauveria bassiana
Parasitic on many insect pests including whiteflies
Also known as "Sugar Icing Fungus“
Causes whitefly mortality ranging from 75% to 80%
80% control of whitefly nymphs has achieved by application of this fungus in irrigated cotton in Imperial Valley of California
Nymph of Bemisia tabaci infested with Beauveria bassiana
Plant Pathology Research Institute, AARI, Faisalabad

48. Entomopathogenic Fungi
Beauveria bassiana
Target pests
Aphids
Whiteflies
Bugs
Mealybugs
Psyllids
Lygus bugs
Chinch bug
Grasshoppers
Thrips
Termites
Fire ants
Flies
Fungal gnats
Shoreflies
Colorado potato beetle
Mexican bean beetle
Japanese beetle
Boll weevil
Cereal leaf beetle
Bark beetles
Strawberry root weevil
Caterpillars
European corn borer
Codling moth
Douglas fir tussock moth
Silkworm
Mites
Plant Pathology Research Institute, AARI, Faisalabad

49. Entomopathogenic Fungi
Metarhizium
Metarhizium anisopliae, formerly known as Entomophthora anisopliae, is a fungus that grows naturally in soils throughout the world
Disease caused by this is called “green muscardine” disease because of the green colour of spores
Cause whitefly mortality more than 75%
Plant Pathology Research Institute, AARI, Faisalabad

50. Entomopathogenic Fungi Metarhizium anisopliae Metarhizium flavoviride
Target pests
Termites
Thrips
Mosquitoes
Locust
Diamondback moth
Mites
Chinch bug
Grasshoppers
Fire ants
Flies
Fungal gnats
Shoreflies
Whiteflies
Metarhizium anisopliae
Metarhizium flavoviride
Plant Pathology Research Institute, AARI, Faisalabad

51. Entomopathogenic Fungi Conidiobolus coronatus
Usually present in soil and on decomposing leaves.
And can cause approximately 95% mortality of B. tabaci adults
Plant Pathology Research Institute, AARI, Faisalabad

52. Entomopathogenic Fungi
Conidiobolus coronatus can cause diseases in animal s & human beings.
Human infections are usually restricted to the rhinofacial area
So care must be taken in dealing with this fungus as entomopathogenic fungus of whitefly
Plant Pathology Research Institute, AARI, Faisalabad

53. Mass production of Entomopathogenic fungi
Many biocompanies in Europe, Asia, America, Holland, Singapore, India etc. producing a series of fungal products based on
Metarhizium,
Beauveria,
Isaria (formerly Paecilomyces),
Paecilomyces lilicenus and
Lecanicillium (formerly Verticillium).
Plant Pathology Research Institute, AARI, Faisalabad

54. Mycoinsecticides available for Bemisia tabaci control
Fungus
Product
Indication
Company
Country
Beauveria bassiana
BotaniGard
Whitefly (Greenhouse)
Emerald BioAgriculture Corporation
USA
Ago Biocontrol Beauveria
Whitefly (Greenhouse, field)
Ago Biocontrol
Colombia
Bea-Sin
Whitefly (Greenhouse, field)
Agrobiologicos del Noroeste S.A. de C.V.
Mexico
Boveril PM
Whitefly
(Greenhouse)
Itaforte BioProdutos
Brazil
Prabhabassiana
Foliar feeding pests, hairy caterpillars of vegetables & fruits plants and sucking pests
Prabhat Fertilizer & Chemical Works
India -
Mycosphere
Singapore
Paecilomyces fumosoroseus
(Isaria fumosorosia)
PreFeRal
Greenhouse, field
Thermo Trilogy/Biobest N.V.
Belgium
Pae-Sin
Greenhouse, field
Paecilomyces lilicenus
Prabhapaecilo.
Plant Pathology Research Institute, AARI, Faisalabad

55. Mycoinsecticides available for Bemisia tabaci control
Fungus
Product
Indication
Company
Country
Verticillium lecanii
(Lecanicillium muscarium)
Ago Biocontrol Verticillium
White fly (Greenhouse)
Ago Biocontrol
Mexico
Mycotal
Whitefly (Greenhouse, field)
Koppert Biological Systems
Holland
Prabha Verti -
Prabhat Fertilizer & Chemical Works
India
Metarhizium anisopliae
(Mycelial fragments & spore )
white grubs, beetle grubs caterpillars, semiloopers, cutworms, sucking pests like Pyrilla, WBPH, mealy bugs , whiteflies
Mycosphere
Singapore
Prabhameta
Fungi mixture:
Lecanicillium Lecanii, Beauveria bassiana and Metarhizium anisopliae.
Asean VBM Bio V
All soft body insects like aphids, mealy bugs, whiteflies, thrips, fruit bores of vegetable crop, white grub / root grubs and Mango hoppers.
Asean Agritechnologies
Plant Pathology Research Institute, AARI, Faisalabad

56. Different Formulation
Mycoinsecticides
Different Formulation
Dustable Powder
Wetable Powder
Emulsifible Concentrate
Suspension Concentrate; Flowable agent
Oil solution
Plant Pathology Research Institute, AARI, Faisalabad

57. Mycoinsecticides Features:
Formulated products, contain materials such as
oils,
humectants,
UV-protectants,
and nutrients
to stimulate germination and growth, have great potential to provide better, more consistent results.
For example,
oil improve adhesion and spreading of spores on the hydrophobic insect cuticle.
Spores may be carried by oil into microhabitats in the insect host or host plant where they are protect from wind, rain, solar radiation, and other environmental stress.
Conidia of fungi formulated in pure vegetable oil and adjuvant oils are claimed to be more resistant to UV radiation than unformulated conidia.
Plant Pathology Research Institute, AARI, Faisalabad

58. Use of entomopathogens:
To summarize…
Use of entomopathogens:
One of the important approaches in Biological management (Ecological management) of important pests in crops.
Maintain the ecological balance of nature
Important component in organic farming for sustainable agriculture
Special tool/weapon to minimize the indiscriminate use of synthetic pesticides
Thus, a great contribution to address the problem of environmental pollution hence, climate change
Plant Pathology Research Institute, AARI, Faisalabad

59. Biological control may give a significant
Global warming…?
Biological control may give a significant
contribution to solve the problem…
Plant Pathology Research Institute, AARI, Faisalabad

60. A great TO ALL OF US… CHALLENGE
Plant Pathology Research Institute, AARI, Faisalabad

61. MUST ACCEPT THE CHALLENGE…
Its duty of
scientists /
researchers
whose work is related to
Plant Pathology,
Agri. Entomology and
Agriculture.
MUST ACCEPT THE CHALLENGE…
Plant Pathology Research Institute, AARI, Faisalabad

62. WHEN? IF NOT TODAY… TOMMOROW ? NEXT WEEK ? NEXT MONTH ? NEXT YEAR ?
NEXT DECADE ?
Maybe its now the right time, we work hand in hand together …to
Plant Pathology Research Institute, AARI, Faisalabad

63. Save the earth before its too late…
Plant Pathology Research Institute, AARI, Faisalabad

64. Thanks for your attention!
Plant Pathology Research Institute, AARI, Faisalabad