1. Cendawan Entomopatogen

2. Insect pathogenic fungi
What are they?
Why study them?
There is good evidence that the Chinese and Romans were aware of insect fungi over 2000 years ago. Japanese seri culturists knew of insect diseases of silkworms over years ago
Insect Fungi have been studied seriously by Western science for about 300 years
Father Jose Torrubia produced this picture of dead insects developing into ‘trees’ following a visit to Cuba in 1749

3. SILKWORM CULTIVATION
Diseases of silkworms are were known to the Japanese over 1000 years ago
By the sixteenth and seventeenth century silkworm production in France and Italy was being ravaged by ‘muscardine’
‘muscardine’ derives from the French word for the sugar-coated almond sweets which the mummified insect larvae resemble

4. AGOSTINO BASSI Born September 25, 1773-Italy
In 1807 he began an investigation of the silkworm disease mal de segno (commonly known as ‘muscardine’), which was causing serious economic losses in Italy
– 10 years before Louis Pasteur, Bassi found disease-causing microorganism was contagious
Died February 8, 1856-Italy

5. Historical aspects Usage traced back ca. 2000 years.
Repoted during the Q’ing Dynasty
Reaumur 1726-Paris
Formally described by Berkeley as Sphaeria sinensis in 1843
Transferred to Cordyceps sinensis by Saccardo in 1870

6. Worm or plant ?
The Insect fungi
good ones
bad ones
EARLY WORK
SAW Beauveria bassiana
AS A BAD GUY
Why study them? Insects fungi as herbal medicines
WHERE, WITHIN THE FUNGAL KINGDOM ARE INSECT FUNGI FOUND?

7. ORDERS CONTAINING INVERTEBRAE FUNGI
Ascomycota
Hypocreales -ca spp.
Eurotiales -<5 spp.
Laboulbeniales -ca spp.
Dothideales -<5 spp.
Basidiomycota
Septobasidiales -ca. 200 spp.
Zygomycota
Zygomycetes -ca. 250 spp.
Trichomycetes -ca. 200 spp.

8. How to find insect fungus and it allies
Looking close to rivers high humidity
Searching the Leaf Litter
Cordyceps emerging from leaf litter
Insect fungi are also found under leaves

9. Not all insect are infected by fungi
The Invasion Process
How Fungi Get In
How they kill
How They Get Out
Not all insect are infected by fungi
Tse-tse fly
flea
Ticks

10. Coleoptera Diptera Hemiptera Spider Orthoptera lepidoptera Hymenoptera
Aphids
Hopper and cicada
White fly
True bug
Spider
Orthoptera
lepidoptera
Hymenoptera

11. The insect cuticule
The whole body surface puls the internal lining of the fore gut and the hind gut and trachea of all insect are line wih cuticule which provides a chemical and physical barrier to infection by micro-organism as well as skeletal support for muscle function

12. The insect cuticule Cuticular Proteins
The main componen of cuticle is chitin bound protein matrix
Apart from insect chitin is produced by other invertebrates and fungi
Cuticular Proteins
The Glue for the chitin
Types of cuticular protein include:
Anthropodin- untanned
Sclerotin-sclerotised
Resiin- rubber like

13. The invasion Process

14. The invasion Process

15. The invasion Process

16. Secondary conidium of Erynia conica with apresorium

17. Glycoprotein Haemolymph
Insect have an open circulatory system with a dorsal heart
Haemolyph function as both as blood and lymph
Blood transport nutrients/ hormones and deals with waste metabolites
Blood is greeenish since there is no haemoglobin
Tissue are bathed with blood
Glycoprotein
Present in haemolyph and other tissue
Function as
Storage molecule
Enzyme
Protection for constituent monosaccharides from enzyme attack

18. Haemolymph nutrients for fungal growth
Trehalose (non-reducing disaccharide of glucose) is the most dominant sugar in insect
Other trehalose using organism
Bacteria
Algae
Fungi
Nematodes
Annelids
Crustaceans
Trehalose (is synthesised from glocise or fructose
Important reserve –readily hydrolized to glucose
High concentration in haemolymph range from 8-60 mg/ml depending on
Species
Developmental stage
sex

19. How does host death occur
Starvation
The insect feeds but fungal takes
Physical disruption of internal tissue and external tissue (prior to death)
Intoxication
Fungal secondary metanolites- destruxin
Auto intoxication
Insect produce toxic metabolites

20. SEM of Aschersonia Enzyme going in- muscle coming out

21. Major Groups Containing Insect Fungi
Ascomycota
Hypocreales -ca sps
Eurotiales -ca. 5 spp
Laboulbeniales -ca spp
Dothideales -ca. 5 spp
Basidiomycota
Septobasidiales -ca. 200 spp
Zygomycota
Zygomycetes -ca spp
Trichomycetes -ca. 200 spp

22. CORDYCEPS SINENSIS (Berk) Sacc. Endosclerotium of Cordyceps
From Yaks and Emperors to the Populace: Now a popular herbal medicine
Fruiting body
Products of Cordyceps sinensis
Caterpilar
dong chong xia cao
‘winter worm, summer-grass’
Endosclerotium of Cordyceps

23. Distribution of Cordyceps sinensis Yearly Harvest (estimated
Tibet
Sichuan
Gansu
Hubei
Zhejiang
Shanxi
Guizou
Qinghai
Yunnan
No Bhutan
No Nepal
Yearly Harvest (estimated
5000 kg
US$ a kg
Each stroma ca. 0.3 g
Ca stromas

24. Medicinal uses of Cordyceps sinensis
Fatigue
Night Sweating
Male and Female hyposexualities, including impotence
Hyperglycemia
Hyperlipidemia
Asthenia after severe illness
Respiratory Diseases
Renal Disfunction and Renal failure
Arrthymias and other heart disease
Liver disease

25. Cordyceps 400+ species
Anamorph: ‘Acremonium’, Akanthomyces, Beauveria, Hirsutella, Hymenostilbe, Isaria, Metharizium, Nomuraea, Polycephalomyces and Verticillium
Host: Coleoptera, Diptera, Homoptera, Hymetoptera, Isoptera Lepidoptera, Neuroptera and Orthoptera

26. Temperature regime in May 2002
Night time -5 to -10C
Sunrise 9 (07.00 to 08.00): -3 to 2C
Late morning ( ): 12 to 22C
Late afternoon (14.00 to 17.00) 2 to 5C
Early evening (18.00 to 21.00): -5 to 0C

27. The Many Anamorphs of Cordyceps
BIOCONTROL
Is the science/art of intentionally enhancing the activity of a beneficial species to reduce the damaging effects of a pest spaces
Akanthomyces
Nomuraea
Beauveria
Paecilomyces
Desmidiospora
Paraisaria
Hirsutella
Pochonia
Hymenostilbe
Polycehalomyces
Isaria
Sorosporella
Lecanicillium
Syngliocladium
Microhilum
Tolypocladum
It is convenient to divide invertebrate pathogenic fungi into two broad categories
General, opportunistic pathogens:
Metarhizium
Beauveria
Fastidious pathogens
Most Cordyceps and the anamorphs

28. Stromatal form in Cordyceps
The Ascomata develop within or on a stroma
Within-immersed
On-superficial

29. Cordyceps Perithecia A. superficial B-D Pseudoimmersed
E. Ordinal immersed
F. Oblique immersed
Ascospore
G. Non disarticulating
H-J Disarticulating

30. Phytocordyceps - pseudoimmersed
Cordyceps on cicada ordinal imersed
Phytocordyceps - pseudoimmersed
Cordyceps nutans oblique immersed

31. The life cycle of Cordyceps militaris
Stromata
Conidiophore
Somatic hypa
Conidia
Asexual Cycle
Sexual Cycle
Lecanicillium
C. militaris
Infection
Germination of ascospore
Germination of conidia
ascus
ascospore

32. Cordyceps unilateralis ant gripping the midvein
Cordyceps khaoyainensis
Cordyceps iranginensis

33. Cordyceps robertsii Cordyceps sinensis New Zealand Maori aweto Asma
A source of charcoal as pigment for tamocco
antibiotics
Cordyceps sinensis
Cordycepic acid
D-mannitol
Cordycepin acid
3’-Deoxyadenosine
Cordyceps militaris
Ergosterine
Ergosterol
Glutamic acid
Fenilalanin
Viagra for women
Proline
Histidine
Valine
Oxivaline
Arginine
Viagra
Alanine
Vitamin B12
And Polysaccharides

34. CLAVICIPITACEOUS FUNGI COMMON IN AGRICULTURAL ECOSYSTEMS
Metarhizium anisopliae -wide host range
Beauveria baasiana –wide host range
Normuraea rileyi –exclusive to Lepidoptera
Hirsutella citriformis - exclusive to hoppers
Hirsutella thompsonii –exclusive to mites
With 400 taxa, not every insect pathogen has potential
Hymenostilbe ventricosa-versus cockroach

35. Hosts of Metarhizium in Thailand
Beauveria bassiana and Metarhizium anosopliae
Hosts of Metarhizium in Thailand
Coleoptera
Lepidoptera
Hemiptera
Leafhoppers
Planthoppers
Cicadas
Diptera
Orthoptera
Araneae

36. Hypocrella schizostachyi
Anamorph
Aschersonia
Host Homoptera
Aleyrodidae and Coccidae
No anamorph known in the nature
Pathogen can end up 1000x more massive then host
Hypocrella schizostachyi

37. Torrubiella Akanthomyces on Spider
Anamorph: Akanthomyces, Gibellula, Granulomanus “Paecylomyces”
Pseudogibellula Tilachlidium and Verticillium
Host: Diptera, Homoptera, Hymetoptera, Lepidoptera and Spiders
Torrubiella
Akanthomyces on Spider

38. Sterols Polycyclic Aromatics Terpenes
Fungal Secondary Metabolic
Epichloe typhina
Cordyceps sinensis
Lecanicillium lecanii
Tolypocladium koningii
Tolypocladium inflatum
Polycyclic Aromatics
Cordyceps unilateralis
Antimalarial naphthoquinones
Paecylomyces carneus
Anthrotoquinones
Cordyceps pseudomilitaris
Bioxanthracenes
Hypocrella bambusae
Hypocrelins
Terpenes
Epichloe typhina
Cephalosporium aphidicola
Cephalosporium caerulens
Paecylomyces inflatus
Isaria c.f javanica
Aschersonia aleyrodis

39. Cyclosporin-Cyclicundecapeptide
Destruxines
Cyclodepsipeptides
Beauveria bassiana
Beauvericine
Isaria tenuipes
Isaria fumosoroseus
Insecticidal activities
Antibacterial gram +
Antifungal
Metharizium anisophliaes
Aschersonia sp.
Insecticidal
Fitotoxic
Antiviral
Sitotoxic
Imunodepresan
Cyclosporin-Cyclicundecapeptide