Efficiency of a nematode (Efficiency of a nematode (SteinernemaSteinernema carpocapsaecarpocapsae) ) and its symbiotic bacterium (and its symbiotic bacterium (Xenorhabdus Xenorhabdus

nematophilanematophila) at eliminating the termite ) at eliminating the termite CoptotermesCoptotermes curvignathuscurvignathus that infests Para rubber that infests Para rubber

ChakraphanChakraphan HiranwrongweraHiranwrongweraPatcharaponPatcharapon AdisettakulAdisettakulSupathepSupathep TansirichaiyaTansirichaiya

ThailandThailand Agricultural country

Para rubber

Problem from termiteProblem from termite

Chemical controlChemical control Biological controlBiological control

Para RubberPara Rubber Coptotermes curvignathus

Damage from

Coptotermes curvignathus to Para rubbers’ root

Biological controlBiological control

VirusVirusBacteriaBacteriaFungiFungi

NematodeNematode

TemporaryTemporary PermanentPermanent

For the life cycle of nematode , there is only one stage to eliminate the insects: Infective

Juvenile (IJ).

The most efficiency of eliminating insects is The most efficiency of eliminating insects is Steinernema carpocapsae.Steinernema carpocapsae.

There is, in nematode intestine,

symbiosis bacteria Xenorhabdus nematophila.

This bacteria produces some substances that can

eliminate insects.

Developing methods to eliminateDeveloping methods to eliminateCoptotermes curvignathusCoptotermes curvignathus by using nematode by using nematode

To study efficiency of To study efficiency of nematode and bacterianematode and bacteria

from the from the entomopathogenic entomopathogenic

nematode to eliminate nematode to eliminate termite in para rubbertermite in para rubber

Para rubber

Research the information

Preparation

Inject suspension on termite

PreparationPreparation

Termite Coptotermes curvignathus

Nematode Steinernema carpocapsae

Bacteria Xenorhabdus nematophila

Suspension of nematode and bacteria

Survey the rubber plantationSurvey the rubber plantation to findto findthe termitesthe termites Coptotermes curvignathusCoptotermes curvignathus

Collect the sample of termitesCollect the sample of termites

Feed the termites Feed the termites to adapt themselvesto adapt themselves

Divide 30 termites to each plate

Preparation of termite Coptotermes curvignathus

Spray the nematodesSpray the nematodes toto infect maggotsinfect maggots

Wait until the nematodesWait until the nematodesseparate from maggotsseparate from maggots

Keep in Keep in the sponges at 5 cthe sponges at 5 c

Preparation of nematode Steinernema carpocapsae

Adjust the concentrateAdjust the concentrate

Count the number Count the number of the nematodesof the nematodes

Squeeze the sponges which have the nematode Squeeze the sponges which have the nematode insideinside in 0.4% formaldehydein 0.4% formaldehyde

Preparation of nematode Steinernema carpocapsae

Culture the bacteria in NBTA Culture the bacteria in NBTA by using liquid from maggotsby using liquid from maggots

AfterAfter 24 hours move bacteria into YS-broth24 hours move bacteria into YS-broth

Wait until Wait until the maggotsthe maggots diedie

Spray the nematodesSpray the nematodes SteinernemaSteinernema carpocapsaecarpocapsaetoto infect maggotsinfect maggots

ShakeShake forfor 24 hours24 hours

Preparation of bacteria Xenorhabdus Nematophila

Centrifuge bacteriaCentrifuge bacteriain the Appendrop in the Appendrop

Remove a half of mediaRemove a half of media

mix it by vortexmix it by vortex

Put nematodes into Put nematodes into AppendropAppendrop

Mix itMix it

Preparation of suspension of nematode and bacteria

ExperimentExperiment Treatment 1, water

Treatment 2, YS-broth

Treatment 3, formaldehyde

Treatment 5, nematode Steinernema carpocapsae

Treatment 4, bacteria Xenorhabdus Nematophila

Treatment 6, suspension of nematode and bacteria

PreparePrepare

Water

YS-broth

Formaldehyde

NematodeBacteria

Nematode and bacteria

Experiment

Control groupsControl groups Experimental groupsExperimental groups

PipettesPipettes

WaterYS-broth

Formaldehyde

NematodeBacteria

Nematode and bacteria

- ---- Termite- ---- Termite Coptotermes curvignathusCoptotermes curvignathus ---- ----- -

Experiment

AfterAfter 2424 hourshours

Observed and recordedObserved and recorded

Experiment

0

5

10

15

20

25

30

35

1 2 3 4 5 6 7 8 9Treatment

Num

ber

of M

orta

lity

Nematode Bacteria

Mix Control

ys broth formaline

0102030405060708090100

Nematode Bacteria Mix Water YS-broth Formaldehyde

Condition

Percentage o

f mor

tality

( %

)

0102030405060708090100

Nematode Bacteria Mix Water YS-broth Formaldehyde

Condition

Percentage o

f mor

tality

( %

)

The nematode has the most efficiencyThe nematode has the most efficiencyin eradicating termites in para rubberin eradicating termites in para rubber

Andrew, H.H. Wong and J. Kenneth Grace. (2004). Laboratory Evaluation of the Formosan Subterranean Termite Resistance of Borate-treated Rubberwood Chipboard. University Malaysia Sarawak Faculty of Resource Science and Technology, Malaysia

Blackburn, M., E. Golubeva, D. Bowen, and R. H. ffrench-Constant. (1998). A novel insecticidal toxin from Photorhabdus luminescens, Toxin complex a (Tca), and its histopathological effects on the midgut of Manduca sexta. Applied and Environmental Microbiology. 64:3036-3041.

Daborn, P. J., N. Waterfield, C. P. Silva, C. P. Y. Au, S. Sharma, and R. H. ffrench-Constant. (2002). A single Photorhabdus gene, makes caterpillars floppy (mcf), allows Escherichia coli to persist within and kill insects. Proceedings of the National Academy of Sciences. 99:10742-10747.

Hayat B. , Abdus S. K., Abid F. , Alam Z. and Amanullah K. (2005). Toxic effects of palpoluck Polygonum hydropepper L. and Bhang Cannabis sativa L. plants extracts against termites Heterotermes indicola (Wasmann) and Coptotermes heimi (Wasmann) (Isoptera:Rhinotermitidae). Songklanakarin J. Sci. Technol., 27(4): 705-710

Finally, this project is accredited to the supports and assistances from – The Thailand Research Fund – Plant Protection Research and Development

Office , Department of Agriculture ,Thailand– Mahidol Wittayanusorn School, Thailand