Types of Entomopathogenic Fungi like Verticillium lecanii Beauveria bassiana Nomuraea rileyi Metarrhizium anisopliae(detailed procedure of mass production in bio control lab)

1. DEPARTMENT
OF
AGRIL.ENTOMOLOGY

3. GENERAL INFORMATION OF THE
STUDENT
NAME OF THE STUDENT :
Solunke Balram D
REGISTRATION NO : AKN-108/08

4. MASS PRODUCTION OF
Metarhizium anisopliae
(Deuteromycota; Hyphomycetes)

5.  An entomopathogenic fungus is fungal species that
can act as a parasite of insects and kills or seriously
disables them. These fungi usually attach to the
external body surface of insects in the form of
microscopic bodies (usually asexual, mitosporic spores
also called conidia).

6.  Verticillium lecanii
 Beauveria bassiana
 Nomuraea rileyi
 Metarrhizium anisopliae

7.  Metarrhizium anisopliae is a widely distributed soil-
inhabiting fungus. The spore of M. anisopliae can be
formulated as dust and sprayable formulation. It is
used to control termites, mosquitoes, leafhopper,
beetles etc.

11. rhinoceros beetle
Oryctes rhinoceros

12. The rhinoceros
beetle, Oryctes
rhinoceros is one of
the serious and
important pests of
coconut and has a
wide distribution
and persistent
occurrence in all the
coconut growing
areas in the country.
rhinoceros
beetle

13. • The adult beetle cause severe damage to the growing palms
by feeding on the tender fronds and crowns and resulting in
stunting of the trees. The damage to the spathe results in the
loss of nuts. Young seedlings are sometimes killed outright.

14.  The chemical control measures
adopted against this pest are always
costly, tedious and have to be repeated
 Hence an easy method utilizing the
safe and specific fungus, Metarhizium
anisopliae for the management of this
pest is aimed at.

17. The bottles are inoculated with 1 ml suspension
containing spores of the fungus with the help of a
sterile injection syringe.
Before inserting the needle within the sterile
bottles for drawing spore suspension for
inoculation, the needle of the syringes and collar
region of the bottles are flamed (Bunsen burner).
Coconut water (40 ml) contained in 375 ml side
wise flat liquor bottles plugged with cotton plug
are sterilized in batches of 9-10 bottles in 12 liters
pressure cooker for 20 minutes at 15 psi.

19. The bottles are inoculated in a laminar flow chamber and
rested on flat surface for 2 days or till the surface of
medium is fully covered by the olive green sporulated
fungus.
The whole culture is crushed thoroughly in an ordinary
mixer and used in the field. From a single average size
coconut 5-6 bottles of cultures can be made.

20. The flasks are incubated at room temperature. The spores can be harvested in a
fortnight.
From a clean uncontaminated mother culture in slant loopful quantities of M.
anisopliae spores are transferred aseptically.
The flasks are allowed to cool and taken to laminar flow chamber for inoculation
The conical flasks are plugged with cotton and autoclaved for 20 min at 15 psi.
Carrot cut into small pieces (40 g) is washed in potable water and transferred to
conical flask (250 ml) and 15 ml of distilled water is added.