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MARILYN M. BALAIS
LEIRA JIM V. TANGIAN
MYRTLE ANNE FAYE N. VIRATA
ENTOMOPATHOGENIC NEMATODES
AGAINST Ostrinia furnacalis
(...
INTRODUCTION
Background of the Study
Insects play important parts in the
ecosystem
Entomopathogenic nematodes (EPNs)
as biopesticide ...
About A. Grisella (lesser wax moth)
Wax moth is a serious pest in
beehives and can cause
substantial losses of
combs, dam...
Larval stage
GROUP OF NEMATODES!!!
THE GOOD THE BAD THE UGLY
Entomopathogenic nematodes (EPN
Entomopathogenic nematodes
Entomopathogenic nematodes
Ecological indicators/Pollution
studies
Species Interaction studies
Biological Control Agents
Genetics studies
Molecular s...
Why BIOLOGICAL CONTROL
Agents?
NO TO EXTENSIVE
SYNTHETHIC
CHEMICAL
APPLICATION
 Entomopathogenic Nematodes (EPNs) in the genera Heterorhabditis
and Steinernema are used as biological control agent (BC...
Objectives of the Study
The study aims to apply isolated EPNs from
soil samples from PSHS – CMC grounds
against the Asian ...
Objectives of the Study
3. apply the extracted EPNs to the larva of the corn
borer;
4. note the mortality rate and qualita...
Significance of the Study
EPNs as biopesticide and growth control
of insects
Cheap and easy way to obtain EPNs
Introduc...
Life-cycle of EPNs in vivo and in vitro
Culturing of
EPNs
In the PHILIPPINES?
Recent studies are undergone in MSU-IIT
and now in PSHS-CMC
Corn borers are one of the major pests in
the corn production industry.
Eliminating pests from corn can make a
great imp...
Scope and Limitations of the Study
Usage of the lesser wax worms as bait for
the EPNs from the soil sample
Pathogenicity...
Scope and Limitations of the Study
Only one site is selected within the
PSHS-CMC campus
Collected EPNs will be used
agai...
MATERIALS AND METHODS
Materials and Methods Flow Chart
Wax Worms
Subculture
Pathogenicity
Test on Corn
Borers
Nematode
Subculture
Collection of
...
How to obtain Dauer Juveniles
from the Soil?
Ten-points Plotting
Ten points represented by circle with 1
meter distance in each point will be
made in the sampling area
Quartering Method
Nematode Baiting
 After soil collection from different areas, collect
subsamples and place in a small cup (100g soil only...
White Trap
Subculture of EPNs in
RINGER's solution
Apply
subcultured
EPNs
against the
Corn Borers
Application of the Isolated EPNs
against Corn Borer
 last stage of the corn borer larva was used and small
plastic cups w...
24-well plate
 The same
procedure will
be done for the
lesser wax
Gathering of
Data
 checked after 12, 24, 36, 48 and 72
hours to see if the corn borers were
dead
 Death of larva is caus...
Analysis of Data
 Corn Borer as bait
 The t-test computed the permutation t-test
p-value
 t-test had been acquired usin...
Results and Discussion
 Corn Borer as bait
 cadavers of the corn borer larva cup were then collected
and recorded on the...
Figure 1. Photo of EPNs seen under the
dissecting microscope (magnification 40X)
(taken on September 15, 2012, MSU - IIT).
Results and Discussion
 Pathogenicity test
 Abbot’s formula
 n = Insect population, T = treated, Co = control
 Other o...
Conclusions and Recommendations
 corn borer larva can be used as an alternative
EPN bait
 corn borers are effective bait...
Thank you for listening!
We are now ready for your
questions.
Entomopathogenic nematodes against ostrinia furnacalis (asian corn borer)
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Entomopathogenic nematodes against ostrinia furnacalis (asian corn borer)

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Entomopathogenic nematodes against ostrinia furnacalis (asian corn borer)

  1. 1. MARILYN M. BALAIS LEIRA JIM V. TANGIAN MYRTLE ANNE FAYE N. VIRATA ENTOMOPATHOGENIC NEMATODES AGAINST Ostrinia furnacalis (ASIAN CORN BORER)
  2. 2. INTRODUCTION
  3. 3. Background of the Study Insects play important parts in the ecosystem Entomopathogenic nematodes (EPNs) as biopesticide to control the Corn borers Achroia grisella (Lesser Wax Moth) as alternative EPN bait
  4. 4. About A. Grisella (lesser wax moth) Wax moth is a serious pest in beehives and can cause substantial losses of combs, damage to beehive material and spoil beehive products. 10 to 13 mm long
  5. 5. Larval stage
  6. 6. GROUP OF NEMATODES!!! THE GOOD THE BAD THE UGLY Entomopathogenic nematodes (EPN
  7. 7. Entomopathogenic nematodes
  8. 8. Entomopathogenic nematodes
  9. 9. Ecological indicators/Pollution studies Species Interaction studies Biological Control Agents Genetics studies Molecular studies
  10. 10. Why BIOLOGICAL CONTROL Agents? NO TO EXTENSIVE SYNTHETHIC CHEMICAL APPLICATION
  11. 11.  Entomopathogenic Nematodes (EPNs) in the genera Heterorhabditis and Steinernema are used as biological control agent (BCA) for diverse insect pests. Family : Steinernematidae with 45 known Steinernema sp. Family : Heterorhabditidae with 11 known Heterorhabditis sp. The enemy of my enemy is my friend THE AMBUSHERS THE CRUISERS EPN pest
  12. 12. Objectives of the Study The study aims to apply isolated EPNs from soil samples from PSHS – CMC grounds against the Asian Corn Borer (Ostrinia furnacalis). Specifically, the study aims to: 1. extract the EPNs present from the wax moth larva; 2. identify the EPNs up to its species level;
  13. 13. Objectives of the Study 3. apply the extracted EPNs to the larva of the corn borer; 4. note the mortality rate and qualitative morphology descriptions of the effects of the EPNs on the Asian Corn Borer larvae;
  14. 14. Significance of the Study EPNs as biopesticide and growth control of insects Cheap and easy way to obtain EPNs Introduces a new and environmental – friendly way to eliminate plant pests
  15. 15. Life-cycle of EPNs in vivo and in vitro Culturing of EPNs
  16. 16. In the PHILIPPINES? Recent studies are undergone in MSU-IIT and now in PSHS-CMC
  17. 17. Corn borers are one of the major pests in the corn production industry. Eliminating pests from corn can make a great impact in the Philippine agriculture
  18. 18. Scope and Limitations of the Study Usage of the lesser wax worms as bait for the EPNs from the soil sample Pathogenicity test in the laboratory is limited to the counting of the number of dead Asian Corn Borer larvae, as well as on noting on the qualitative effects of the EPNs on the external morphology of the
  19. 19. Scope and Limitations of the Study Only one site is selected within the PSHS-CMC campus Collected EPNs will be used against the larvae of the corn borer in the laboratory
  20. 20. MATERIALS AND METHODS
  21. 21. Materials and Methods Flow Chart Wax Worms Subculture Pathogenicity Test on Corn Borers Nematode Subculture Collection of Soil Samples Quartering Method Nematode Baiting Artificial Media Preparation Analysis of Data
  22. 22. How to obtain Dauer Juveniles from the Soil?
  23. 23. Ten-points Plotting Ten points represented by circle with 1 meter distance in each point will be made in the sampling area
  24. 24. Quartering Method
  25. 25. Nematode Baiting  After soil collection from different areas, collect subsamples and place in a small cup (100g soil only)  Put at least 5 larval stage of A. grisella inside the cup, cover using the lid and put the sample under room temperature.  After 24 hours, examine the larvae in the soil, if morbid leave for another 24 hrs and if found dead after 3-7 days depending on the species, collect the cadaver  Rinse the cadaver using either distilled water  Transfer the cadaver to white trap
  26. 26. White Trap
  27. 27. Subculture of EPNs in RINGER's solution Apply subcultured EPNs against the Corn Borers
  28. 28. Application of the Isolated EPNs against Corn Borer  last stage of the corn borer larva was used and small plastic cups with filter paper were each filled with one larva  (1 larva/concentration times 3 replicates) times 3 trials  nematodes were counted by putting small drops of the solution on the Petri dish and then the droplets were examined under the spectroscope  add drop/s with 5, 10, 20, 30, 40, 50 EPNs) on each borer  filter paper in the cup is moistened
  29. 29. 24-well plate  The same procedure will be done for the lesser wax
  30. 30. Gathering of Data  checked after 12, 24, 36, 48 and 72 hours to see if the corn borers were dead  Death of larva is caused by successful invasion of EPNs if the cadaver shows discoloration and presence of EPNs when dissected  Note on the mortality rate per
  31. 31. Analysis of Data  Corn Borer as bait  The t-test computed the permutation t-test p-value  t-test had been acquired using (PAST) software  Pathogenicity Test  Probit analysis  Abbot formula
  32. 32. Results and Discussion  Corn Borer as bait  cadavers of the corn borer larva cup were then collected and recorded on the 3rd, 5th, and 7th day of the baiting procedure  Discoloration  Heterorhabditis group  no significant differences between the permutation t-test values since the results were more than 0.05  Pathogenicity test  LC50 of the pathogenicity test, derived by probit analysis  44% control mortality
  33. 33. Figure 1. Photo of EPNs seen under the dissecting microscope (magnification 40X) (taken on September 15, 2012, MSU - IIT).
  34. 34. Results and Discussion  Pathogenicity test  Abbot’s formula  n = Insect population, T = treated, Co = control  Other observations
  35. 35. Conclusions and Recommendations  corn borer larva can be used as an alternative EPN bait  corn borers are effective bait for Heterorhadbitis group  40 concentration/larva is best effective since it has the highest percent mortality from the LC50 as well as in the Abbot mortality after 12 and 72 hours  suggest that the set-up should be monitored regularly and new insect larva may be applied with the nematodes  recommended that there will more specific and
  36. 36. Thank you for listening! We are now ready for your questions.

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