Prospect of The Entomopathogenic Nematodes As Biological Control For The Future in Indonesia and Asean Countries

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010
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PROSPECT OF THE ENTOMOPATHOGENIC NEMATODES AS BIOLOGICAL CONTROL FOR THE FUTURE IN ASEAN COUNTRIES
Prof.Dr. sc. agr. Ir. Didik Sulistyanto
Prof. Dr. Ir. Didik Sulistyanto
Indonesia 2013
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INTRODUCTION
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Entomopathogenic nematodes have emerged as excellent biological control agents for insect pests, Broad host range, safety to non-target organisms and the environment, Exemption from registration in many countries, ease of mass production and application, ability to search for pests, Rapid host mortality, potential to recycle in the environment and capability with most agricultural chemicals,
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Entomopathogenic nematodes as biological control agents for soil-dwelling stage of many insect pests, Progress EPNs available commercially for large scale application in plantation, horticulture, turf grass, rice plantation, fruit plantation, coffee, cacao, sugarcane, etc, Steinernema sp. and Heterorhabditis sp. isolat effective as biological control insect pests and ease to mass produce in vivo and in vitro culture,
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PENDAHULUAN
POTENTIAL OF ENTOMOPATHOGENIC NEMATODES
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Advantage:
1. Rapid host mortality 24-48 hour,
2. Broad host range, Safety to non-target organisms and environment,
3. Ease of mass production in vitro and in vivo,

4. Ease to application in the field, 5. Compatibility with most agricultural chemicals, 6. Available commercially for large-scale application in plantation and others,
ENTOMOPATHOGENIC NEMATODES
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- Size 500-800 um, - Baktery simbion: Xenorhabdus spp.- dari Steinernema spp. Photorhabdus sp.- dari Heterorhabditis spp. - Efektif on all pest like: -Pest (Lepidoptera, -Coleoptera, Diptera, dll) - Hoticulture, Rice, Fruit, Industrial Crop, Crop Plantation, - Isolated from any kind of soil
Didik Photo Collection
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LIFE CYCLE OF ENTOMOPATHOGENIC NEMATODES
INFECTION MECHANISM OF ENTOMOPATHOGENIC NEMATODES IN INSECT

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Didik Sulistyanto Photo Collection

INSECT MORTALITY WITHIN 24 HOURS

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Nematodes penetrated in anus of P. xylostella MAIN MENU
PATHOGENICITY OF EPNs SYMBIOTIC BACTERIA
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Bakteria
Nematodes
Didik Photo Collection
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MECHANISM OF PENETRATION OF EPNs IN INSECT PESTS

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IDENTIFICATION TECHNIQUE OF ENTOMOPATHOGENIC NEMATODES

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Identification technique with: (a) Infektif Juvenile (IJs),
(b) Adult Male,

(c) Molekuler biology,
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IDENTIFICATION ON INFECTIVE JUVENILE
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HEAD OF HETERORHABDITIS SPP.
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IDENTIFICATION OF HETERORHABDITIS SPP. ADULT MALE
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IDENTIFICATION OF STEINERNEMA SPP. ADULT MALE
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HETERORHABDITIS SPP. ADULT MALE

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HETERORHABDITIS SPP. ADULT MALE

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STEINERNEMA SPP. ADULT MALE

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STEINERNEMA SPP. ADULT MALE

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ENTOMOPATHOGENIC NEMATODES SPECIES
ENTOMOPATOGENIC 2 GENUS:
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(a) Heterorhabditis sp.- Photorhabdus sp.
(b) Steinernema sp.- Xenorhabdus sp.

HETERORHABDITIS SP.:
- H. indica
- H. bacteriophora - H. popilliae
- Heterorabhditis spp.
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STEINERNEMA SP.:
- S. carpocapsae
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- S. affinis
- S. glaseri
- S. bicornutum - S. feltiae - S. japanicum - Steinernema spp.
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SYMBIOTIC OF BACTERIA ENTOMOPATHOGENIC NEMATODES

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Symbiotic bacteria of entomopathogenic
nematodes live in Vesikel, Symbiotic bacteria have 2 phase: primer and secondary phase,
Very difficult to separate with Host,

Very sensitive with Ultra Violet 200 2000 cell of symbiotic bacteria per EPN,
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SYMBIOTIC BACTERIA OF ENTOMOPATOGENIC NEMATODES

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SYMBIOTIC BACTERIA OF Xenorhabdus nematophilus

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SYMBIOTIC BACTERIA 0F Photorhabdus luminenscens
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LIVE
Steinernema sp.
Heterorhabditis sp.
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ORBITAL SHAKER FOR PRODUCTION

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MASS PRODUCTION SCHEME

Strain Collection
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Mass Production
In Vivo Small Scale Media Agar

In Vitro Medium Scale Large Scale Bio-reaktor Liquid Culture 30
INSECT
Media Bedding
MASS PRODUCTION TECHNIQUE OF ENTOMOPATHOGENIC NEMATODES

(1) IN VIVO METHODS (INSECT CULTURE):
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- Tenebrio mollitor,
-Galleria mellonella, - with WHITE TRAP method,
- harvest after 7-10 days,

- on petridis with paper and water, - keep in room temperature and dark avoid the Ultra Violet,
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In-Vivo
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MASS PRODUCTION TECHNIQUE OF ENTOMOPATHOGENIC NEMATODES
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(2) IN VITRO METHODS

- use solid or liquid culture,
- harvest after 7-14 days and filter by 30 um mesh,

- keep in room temperature and dark avoid the Ultra Violet.
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MASS PRODUCTION OF ENTOMPATHOGENIC NEMATODES IN VIVO AND IN VITRO

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IN VIVO
IN VITRO
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PRODUCTION IN VITRO (Sponge)

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MASS PRODUCTION IN FLASK
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MASS PRODUCTION TECHNIQUE OF EPNs

(a) In vivo (b) In vitro in media agar In vitro in solid media (d) Harvest technique
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MASS PRODUCTION EPNS IN BIOREACTOR IN VITRO
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IN VITRO BIOREAKTOR
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Bio-Fermentor For Mass Production of Entomopathogenic Nematodes in Liquid media volume 50.000 100.000 liter
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SIMPLE HARVEST TECHNIQUE OF ENTOMOPATHOGENI C NEMATODES
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FORMULATION TECHNIQUE OF ENTOMOPATHOGENIC NEMATODES

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(a) Sponge, (b) Mineral soil, (c) Mountain soil, (d) Dust, (e) Extract mineral,

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MASS PRODUCTION OF ENTOMOPATHOGENIC NEMATODES FOR FARMERS

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IN VIVO METHOD:
- Serangga di kumpulkan dan diletakkan dalam wadah yang berisi kertas serap air/tanah pasir, - Larva diinokulasi dengan NEP dan diusahakan jangan menggenang (kondisi lembab), - Setelah serangga mati (warna coklat/merah), disusun di wadah terbalik yang berada dalam wadah besar yang dilapisi kertas & berisi air, - Tutup dan biarkan selama 1- 2 minggu dan setelah itu dipanen untuk diaplikasikan,
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ENTOMOPATHOGENIC NEMATODES PRODUCT AS BIOLOGICAL CONTROL OF PESTIN THE WORLD

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PRODUCT OF ENTOMOPATHOGENIC NEMATODES LOCAL ISOLATES FROM INDONESIA
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ENTOMOPATHOGENIC NEMATODES PRODUCT FOR CONTROL HORTICULTURE INSECTS PEST

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ENTOMOPATHOGENIC NEMATODES PRODUCT FOR CONTROL WHITE GRUBS INSECTS PEST

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ENTOMOPATHOGENIC NEMATODES PRODUCT FOR CONTROL COFFEE AND CACAO INSECTS PEST
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ENTOMPPATHOGENIC NEMATODES PRODUCT FOR CONTROL FLIES INSECTS PEST ON CAMPIGNON
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PEMANFAATAN NEP SEBAGAI AGENSIA HAYATI HAMA

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Effektif mengendalikan hama dari ordo
Lepidoptera (kupu-kupu), Coleoptera (kumbang), Diptera (lalat), dan serangga hama lainnya, Aman terhadap musuh alami (parasitoid, predator dan musuh alami lainnya) Kompatibel diaplikasikan dengan pengendalian kimiawi dan hayati lainnya, Aman dimanfaatkan untuk tanaman pangan, sayuran, perkebunan dan jenis tanaman lainnya, Diaplikasikan pada sore hari untuk menghindari sinar matahari (UV).
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Jenis-Jenis Hama Pada Tanaman Pertanian yang Dapat Dikendalikan dengan Nematoda Entomopatogen (Steinernema sp. & Heterorhabditis sp.)
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FIELD RESEARCH IN BROMO MOUNTAIN

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CABBAGE PROTECTED BY ENTOMOPATHOGENIC NEMATODES

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Cabbage Normal
Cabbage destructed by Plutella xylostella and Crocidolomia binotalis

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Plutella xylostella and Crocidolomia sp. Insect Pests Mortality 80 - 90% in the field control with EPNs
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INFECTION OF EPNs ON SEVERAL INSECT PESTS
P. xylostella
(a) TERINFEKSI (b) SEHAT
b
a
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CABBAGE WITH FULL PESTICIDE
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Khob Khun Khrab

Email:
sulis2303@yahoo.com www.didiksulis.blogspot.com
HP : 08124982150
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Prospect of The Entomopathogenic Nematodes As Biological Control For The Future in Indonesia and Asean Countries

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

POTENTIAL OF ENTOMOPATHOGENIC NEMATODES

3. Ease of mass production in vitro and in vivo,

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

LIFE CYCLE OF ENTOMOPATHOGENIC NEMATODES

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

INFECTION MECHANISM OF ENTOMOPATHOGENIC NEMATODES IN INSECT

Didik Sulistyanto Photo Collection

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

INSECT MORTALITY WITHIN 24 HOURS

Didik Sulistyanto Photo Collection

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

Nematodes penetrated in anus of P. xylostella MAIN MENU

Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

PATHOGENICITY OF EPNs SYMBIOTIC BACTERIA

Prof. Dr. Ir. Didik Sulistyanto

Didik Photo Collection

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

MECHANISM OF PENETRATION OF EPNs IN INSECT PESTS

Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

IDENTIFICATION TECHNIQUE OF ENTOMOPATHOGENIC NEMATODES

Identification technique with: (a) Infektif Juvenile (IJs),

(b) Adult Male,

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

IDENTIFICATION ON INFECTIVE JUVENILE

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

HEAD OF HETERORHABDITIS SPP.

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

IDENTIFICATION OF HETERORHABDITIS SPP. ADULT MALE

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

IDENTIFICATION OF STEINERNEMA SPP. ADULT MALE

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

HETERORHABDITIS SPP. ADULT MALE

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

HETERORHABDITIS SPP. ADULT MALE

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

STEINERNEMA SPP. ADULT MALE

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

STEINERNEMA SPP. ADULT MALE

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

ENTOMOPATHOGENIC NEMATODES SPECIES