Monday, June 11, 2007

Strong mosquitocidal Bacillus thuringiensis from Mt. Everest

Strong mosquitocidal Bacillus thuringiensis from Mt. Everest

Upendra Thapa Shreshtha1, Kiran Babu Tiwari1, Anjana Singh2, Vishwanath Prasad Agrawal1*

1Research Laboratory for Agricultural Biotechnology and Biochemistry (RLABB), Universal Science College, Maitidevi, Kathmandu, Nepal; 2Central Department of Microbiology, Tribhuvan University, Kirtipur, Nepal

*Address for Correspondence: Prof. Vishwanath Prasad Agrawal, RLABB, Tel: +977-1-4442775, E-mail: vpa@wlink.com.np

Microbial insecticides are especially valuable as their toxicity to non-target animals and humans is extremely low compared to other commonly used chemical insecticides. They are safer for both the pesticide users and consumers of pesticide treated crops (Neppl, 2000). The soil bacterium Bacillus thuringiensis fulfills the requisites of a microbiological control agent of agricultural pests and vectors that cause massive crop destruction (Ben-Dov et al, 1999). The main target pest of B. thuringiensis insecticides includes various Lepidoptera (butterfly), Diptera (flies and mosquitoes), Coleopteran (Beatle) and some strains of nematodes (Schnepf et al , 1998). The kurstaki HD1 strain of B. thuringiensis is shown to possess wide spread insecticidal properties (Dulmage, 1970).

To study B. thuringiensis population from high altitude, soil samples were collected from Sagarmatha National Park (SNP) and Phereche of Khumbu region situated in the base camp of Mt. Everest, and processed in Research Laboratory for Agricultural Biotechnology and Biochemistry (RLABB). B. thuringiensis strains were isolated from soil samples by acetate selection method (Travers et al, 1987, Shrestha et al. 2006) and identified by standard microbiological techniques including colonial and morphological characteristics, and biochemical tests (Bergey’s Manual, 1986).

Out of 86 δ-endotoxin positive isolates (total 109), 10 randomly selected ones were used for insect bioassay. Larvae, collected from the ditches in local area of Bode, Bhaktapur, Nepal, were reared in a jar containing 100 ml of sterilized water containing 0.3 ml of 5% Brewer's Yeast and 5 ml of B. thuringiensis stationary phase culture and allowed to stand for 3 days. Although all isolates tested were effective against the larvae, isolate S6 was the most effective of all (Fig. 1).

The isolates, obtained from the soil samples above 4000m altitudes where mosquitoes are not expected, may be novel. The S6 isolate showing potent insecticidal property tested against dipterans need to be studied further in larger trials so that it can have applicability to reduce the mosquitoes and different diseases caused by these vectors (Malaria, Filaria, Kalazar etc).

Acknowledgment : We thank CNR of Italy to support this work.


References:

Ben-Dov E, Wang Q, Zaritsky A, Manasherob R, Barak Z, Schneider B, Khamraev A, Baizhanov M, Glupov V, Margalith Y. Multiplex PCR screening to detect cry9 genes in Bacillus thuringiensis strains. Appl Environ Microbiol 1999; 65: 3714-6.

Bergey’s Manual of Systematic Bacteriology, Volume 2, 1986.

Dulmage HT. Production of spore-delta-endotoxin complex by variants of Bacillus thuringiensis in two fermentation media. J Invertebr Pathol 1970; 16: 385-9.

Neppl CC (2000). Managing Resistance to Bacillus thuringiensis Toxins. Environmental Studies University of Chicago.

Schnepf E, Crickmore N, Van Rie J, Lereclus D, Baum J, Feitelson J, Zeigler DR and Dean DH. Bacillus thuringiensis and its pesticidal crystal proteins. Microbiol Mol Biol Rev 1998; 62: 775-806.

Shrestha UT, Sahukhal GS, Pokhrel S, Tiwari KB, Singh A and Agrawal VP. Delta- endotoxin immuno cross-reactivity of Bacillus thuringiensis isolates collected from Khumbu base camp of Mount Everest region. J Food Sci Technol Nepal 2006; 2: 128-131.

Travers RS, Martin PA and Reichelderfer CF. Selective Process for Efficient Isolation of Soil Bacillus spp. Appl Environ Microbiol 1987; 53: 1263-6.

1 comment:

Anonymous said...

How do you justify such a strong mosquitocidal activity of B.t. in a cold climate like Mt. Everest ?