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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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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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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Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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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

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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,
Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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LIFE CYCLE OF ENTOMOPATHOGENIC NEMATODES

Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

INFECTION MECHANISM OF ENTOMOPATHOGENIC NEMATODES IN INSECT


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Didik Sulistyanto Photo Collection


Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

INSECT MORTALITY WITHIN 24 HOURS


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Didik Sulistyanto Photo Collection


Prof. Dr. Ir. Didik Sulistyanto

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

Nematodes penetrated in anus of P. xylostella MAIN MENU

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

PATHOGENICITY OF EPNs SYMBIOTIC BACTERIA

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Bakteria

Nematodes

Prof. Dr. Ir. Didik Sulistyanto

Didik Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

MECHANISM OF PENETRATION OF EPNs IN INSECT PESTS


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Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

IDENTIFICATION TECHNIQUE OF ENTOMOPATHOGENIC NEMATODES


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

(b) Adult Male,


(c) Molekuler biology,

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

IDENTIFICATION ON INFECTIVE JUVENILE

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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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HEAD OF HETERORHABDITIS SPP.

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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IDENTIFICATION OF HETERORHABDITIS SPP. ADULT MALE

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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IDENTIFICATION OF STEINERNEMA SPP. ADULT MALE

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

HETERORHABDITIS SPP. ADULT MALE


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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

HETERORHABDITIS SPP. ADULT MALE


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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

STEINERNEMA SPP. ADULT MALE


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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

STEINERNEMA SPP. ADULT MALE


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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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
Prof. Dr. Ir. Didik Sulistyanto

- Heterorabhditis spp.
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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

STEINERNEMA SP.:

- S. carpocapsae
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- S. affinis

- S. glaseri
- S. bicornutum - S. feltiae - S. japanicum - Steinernema spp.
Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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,

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

SYMBIOTIC BACTERIA OF ENTOMOPATOGENIC NEMATODES


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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

SYMBIOTIC BACTERIA OF Xenorhabdus nematophilus


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Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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SYMBIOTIC BACTERIA 0F Photorhabdus luminenscens

Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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Didik Sulistyanto Photo Collection

LIVE
Prof. Dr. Ir. Didik Sulistyanto

Steinernema sp.

Heterorhabditis sp.
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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

ORBITAL SHAKER FOR PRODUCTION


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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

MASS PRODUCTION SCHEME


Strain Collection
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Mass Production

In Vivo Small Scale Media Agar


Prof. Dr. Ir. Didik Sulistyanto

In Vitro Medium Scale Large Scale Bio-reaktor Liquid Culture 30

INSECT

Media Bedding

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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,

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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In-Vivo

Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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.

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

MASS PRODUCTION OF ENTOMPATHOGENIC NEMATODES IN VIVO AND IN VITRO


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IN VIVO
Prof. Dr. Ir. Didik Sulistyanto

IN VITRO
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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

PRODUCTION IN VITRO (Sponge)


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Didik Sulistyanto Photo Collection


Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

MASS PRODUCTION IN FLASK

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Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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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

Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

MASS PRODUCTION EPNS IN BIOREACTOR IN VITRO

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Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

IN VITRO BIOREAKTOR

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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Bio-Fermentor For Mass Production of Entomopathogenic Nematodes in Liquid media volume 50.000 100.000 liter

Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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SIMPLE HARVEST TECHNIQUE OF ENTOMOPATHOGENI C NEMATODES

Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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,

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

ENTOMOPATHOGENIC NEMATODES PRODUCT AS BIOLOGICAL CONTROL OF PESTIN THE WORLD


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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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PRODUCT OF ENTOMOPATHOGENIC NEMATODES LOCAL ISOLATES FROM INDONESIA

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

ENTOMOPATHOGENIC NEMATODES PRODUCT FOR CONTROL HORTICULTURE INSECTS PEST


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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

ENTOMOPATHOGENIC NEMATODES PRODUCT FOR CONTROL WHITE GRUBS INSECTS PEST


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Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

ENTOMOPATHOGENIC NEMATODES PRODUCT FOR CONTROL COFFEE AND CACAO INSECTS PEST
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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

ENTOMPPATHOGENIC NEMATODES PRODUCT FOR CONTROL FLIES INSECTS PEST ON CAMPIGNON

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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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).

Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

Jenis-Jenis Hama Pada Tanaman Pertanian yang Dapat Dikendalikan dengan Nematoda Entomopatogen (Steinernema sp. & Heterorhabditis sp.)
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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

FIELD RESEARCH IN BROMO MOUNTAIN


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Didik Sulistyanto Photo Collection


Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

CABBAGE PROTECTED BY ENTOMOPATHOGENIC NEMATODES


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Cabbage Normal
Prof. Dr. Ir. Didik Sulistyanto

Cabbage destructed by Plutella xylostella and Crocidolomia binotalis


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Didik Sulistyanto Photo Collection

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

Plutella xylostella and Crocidolomia sp. Insect Pests Mortality 80 - 90% in the field control with EPNs
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Prof. Dr. Ir. Didik Sulistyanto

Didik Sulistyanto Photo Collection

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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INFECTION OF EPNs ON SEVERAL INSECT PESTS

P. xylostella

(a) TERINFEKSI (b) SEHAT

b
Prof. Dr. Ir. Didik Sulistyanto

a
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Didik Sulistyanto Photo Collection

NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

CABBAGE WITH FULL PESTICIDE

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Prof. Dr. Ir. Didik Sulistyanto

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NARESUAN UNIVERSITY-PHITSANULOK- 29 JULI 2010

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Khob Khun Khrab


Email:
sulis2303@yahoo.com www.didiksulis.blogspot.com
Prof. Dr. Ir. Didik Sulistyanto

HP : 08124982150
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