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Mass production of Metarhizium anisopliae (Mets.) dust formulation using Jack fruit seed powder and
testing its pathogenecity against Corcyra cephalonica Stainton
Anitha S, Sam Manohar Das S, Thivya S
Department of Zoology, Scott Christian College (Autonomous), Nagercoil, Tamil Nadu, India.
Abstract
The major objective of this study is to mass produce Metarhizium anisopliae (Mets.) in a cheap substrate like jack fruit seed and to
ascertain spore quality by evluating the effect of their powder formulation against the life stages stored grain pest, Corcyra
cephalonica Stainton (Lepidoptera: Pyralidae). The production of conidiospores in jack fruit seed medium was high. The 16 days
old culture was fully covered with the conidiospores. The conidiospore production recorded after 16 days of incubation was 21.2x
104 spores/ ml and the viable count was 41.4x107 cfu/ ml. The dust formulation of M. anisopliae containing the adjuvants like
charcoal, kaolin, wheat flour and ash was prepared to contain 3, 6 or 9 percent dry fungal powder preparations of different
concentrations were very effective against all the larval instars of C. cephalonica. The mortality rate increased with the increasing
concentration of dust formulation and days of exposure. Third instar larvae were more susceptible to the fungal formulation than the
older ones. The culture media and adjuvants used for the preparation are ideal, since the cost of materials is very low and the
entomopathogenecity high.
Keywords: Conidiospores, Metarhizium anisopliae (Mets), Corcyra cephalonica Stainton, jack fruit seed, viable count
Table 1. Spore quality of M. anisopliae spores produced in the jack fruit seed powder medium after different days of incubation
Spore qulality
Days of
Sl. No Wet biomass Dry biomass (in Conidiospore production Viability of Conidiospore
incubation
(in gm/100ml) gm/100ml) (X104 spores/ml) (X107 cfu/ml)
1 6 7.49± 0.66 0.31±0.08 13.4±1.07 21.4±1.13 (2nd day)
2 8 12.7±1.09 0.52±0.03 18.9±1.09 35.4±2.14 (3rd day)
3 10 20.68±1.14 0.81±0.06 19.7±1.32 38.3±2.19(4th day)
4 12 22.7±1.11 0.97±0.07 20.4±1.33 39.1±1.98(5th day)
5 14 23.9±1.93 1.01±0.07 20.8±1.33 39.8±2.11(6th day)
6 16 25.2±1.96 1.15±0.09 21.2±1.29 41.4±2.14(7th day)
The viable count recorded after 7 days of incubation was rich in amino acids showed stimulatory effect on the growth and
41.4x107 cfu/ml. This high count is presumably due to the germination of conidiospores. Rath et al. [13] examined the
presence of high amount carbohydrates in the jack fruit seed. Li utilization of 49 carbohydrates by 134 isolates of M. anisopliae
and Holdom [12] examined the effects of a range of carbon and and concluded that carbohydrate utilization was a useful and
nitrogen sources and vitamins on colony formation, mycelial biologically relevant taxonomic criteria and tool for the
growth and sporulation of two isolates and concluded that separation of Metarhizium strains and other entomogenous
soluble starch was best among different carbon sources tested fungi.
for growth of M. anisopliae and also found that nitrogen sources
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Mass production of conidiospores of M. anisopliae - Jack Jack seeds that support the luxuriant growth of M. anisopliae,
fruit seed medium are cheap, compared to commercial synthetic culture media.
These cheaper substrates are convenient to handle for large
scale use because labour and energy requirements are low. The
formulation of M. anisopliae conidiospores with ash, charcoal,
chalk powder and wheat flour is found to be very effective in
the control of C. cephalonica larvae and the production cost was
very cheap when compared to other mycopesticides. This
preparation can be kept open in storage areas as ovitraps, so that
mated C.cephalonica females would be attracted to lay eggs
into the mixture resulting in the death of the emerging larvae.
Such ovitraps would considerably reduce the population of C.
cephalonica in ware houses where rice and wheat are stored.
4. Conclusion
Jack fruit seed powder is comparatively cheaper than the other
synthetic media available in the market. This cheap media and
low-tech mass production methods used to supply the fungal
pesticides at an affordable cost. The dry fungal powder
3 days old culture preparations of different concentrations were very effective
against all the larval instars of C. cephalonica.
References
1. Raimbault M. General and Microbiological aspects of solid
substrate fermentation. Electron, J Biotechnol. 1998;
1(3):1-15.
2. Keller S, Kessler P, Schweizer C. Distribution of insect
pathogenic soil fungi in Switzerland with special reference
to Beauveria brongniartii and Metarhizium anisopliae.
Biocontrol 2003; 48:307-319.
3. Wells HD, Bell DK, Jonorski CK. Efficacy of Trichoderma
harzianum as a biocontrol for Sclerotium rolfsii.
Phytopathology 1972; 62:442-447.
4. Kumar A, Anandaraj M, Srinivasan V, Veena S, Sarma
YK. Coconut water amended coir pith- A conductive
medium for mass multiplication of biocontrol agent
Trichoderma spp. spices and Aromatic plants: Challenges
16 days old culture and opportunities in the New Century. Centennial
conference on spices and aromatic plants, 2000, 267-273.
The three formulations (3, 6 and 9% w/w) of M. anisopliae were 5. Gandhikumar N, Raguchander T, Prabhakar K. Mass
tested against the third, fourth and fifth instar C. cephalonica. multiplication of biochemical agents. Annals of Plant
The mortality rate of C. cephalonica larvae increased with the Protection Sciences 2001; 9(1):140-142.
increasing concentration of M. anisopliae formulation and 6. Prakash MG, Gopal KV, Anandaraj M, Sarma YR.
prolonged time of exposure. In 9 percent concentration, Evaluation of substrates for mass multiplication of fungal
10±0.44 third instar larvae died after 9 days of exposure. In the biocontrol agents Trichoderma harzianum and T. virens.
fourth instar larvae, the corresponding mortality rate was Journal of Spices and Aromatic Crops. 1999; 8(2):207-210.
8±0.70 and fifth instar larva, 8±0.44. Which is about 20 percent 7. Lakshmi SM, Alagammai PL, Jayaraj K. Studies on mass
less compared to the third instar larvae (table 2). Batta [14] who culturing of the entomopathogen, Verticillium lecanii on
studied the control of rice weevil with various formulations of three grain media and its efficacy on Helicoverpa
M. anisopliae, found that the mixture of M. anisopliae armigera. In Ignachimuthu S. and Sen, S. (eds.) Microbials
conidiospores with charcoal and oven ash at a rate of 2% or 2.8 in Insect pest management, Oxford and IBH publishing
mg resulted in 73.3- 86.7% mortality of Sitophilus oryzae L. Pvt. Ltd., New Delhi., 2001, 23-27.
after 7 days, when treatments were gives before or after pest 8. Lopez-Liorca LV, Carbonell T. Use of almond mesocarp
infestation. The mortality caused by fungal isolates was for production of the entomopathogenic fungus,
confirmed by external sporulation of mycelia on the insect Verticillium lecanii. Can, J Microbiol. 1998; 44(9):886-
cadavers. 895.
The results also indicated that the third instar larvae were more 9. Farsi MJ, Askari H, Jahromi KT, Pakdel AK. Effect of
susceptible to the fungal formulation than the later instars. This important ecological factors on blastospore production of
may due to the incresed detoxicative metabolism in the mature Verticillium lecanii DAOM 198499 in liquid medium,
larvae. Jbilou et al. [15] reported that early instar larvae of Iranian J Agric. Sci. 2005; 36(1):109-119.
Tribolium castaneum are more prone to four medicinal plant 10. Derakhshan A, Rabindra RJ, Ramanujam B. Effect of
extracts than the final instar and adults. storage conditions of formulation on viability of
3
Verticillium lecanii (Zimmerman) viegas and its virulence
to Brevicoryne brassicae (L), Journal of Biological
Sciences. 2008; 8(2):498-501.
11. Anitha S. Mass production and preservation of
entomopathogenic fungus, Metarhizium anisopliae
(Metchnikoff) for biological control. Ph.D thesis.
Manonmaniam Sundaranar University, Tirunelveli, 2011.
12. Li DP, Holdom DG. Effects of pesticides on growth and
sporulation of Metarhizium anisopliae (Deuteromycotina:
Hyphomycetes), J Inver. Pathol. 1994; 63:209-211.
13. Rath AC, Carr CJ, Graham BR. Characterization of
Metarhizium anisopliae strains by carbohydrate utilization.
Journal of Invertebrate Pathology. 1995; 65:152-161.
14. Batta YA. Control of rice weevil (Sitophilus oryzae L)
(Coleoptrea: Curculionidae) with various formulations of
Metarhizium anisopliae, Journal of crop protection. 2004;
23(2):103-108.
15. Jbilou R, Ennabili A, Rachid FS. Insecticidal activity of
four medicinal plant extracts against Tribolium castaneum
(Herbst) (Coleoptera: Tenebrionidae), African Journal of
Biotechnology. 2006; 5(10):936-940.
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