Insecticidal Virus for effective Codling Moth control.

1. Insecticidal virus
for control of
Codling Moth larvae

2. Active ingredient:
Codling moth granulosis virus
Nucleocapsid
Glycoprotein
envelope
Viral
DNA
Granulin
protein
• Cydia pomonella granulovirus (CpGV)
Occlusion
• Natural pathogen of codling moth body (OB)

3. • Virus is produced in mass-reared codling moth larvae
• Formulated as aqueous suspension concentrate
• Highly specific to codling moth
• No effect on beneficials, wildlife, livestock, or users
• Listed by OMRI and NOP for use in organic production
• Registration in all major apple & pear producing states

4. CpGV Products Registered with the
US EPA (2005)
Product: Cyd-X Carpovirusine ViroSoft CP4
Company Certis USA Sumitomo BioTepp
AI Conc. (OB* per liter) 3.3  1013 1  1013 4  1013
Rate (fl. oz. per acre) 1–6 13.7 3.2
Rate (OB* per acre) 1 – 6  1012 4  1012 4  1012
*OB = occlusion bodies

5. Biology & Mode of Action of CpGV
• Virus must be ingested by larvae to cause
infection
• Once in gut, virus penetrates cells and begins
to replicate
• LD50 is 1 or 2 virus particles per larva
(Cyd-X contains about One Trillion per ounce!)
Fat body of healthy larva
• Modes of transmission:
- Horizontal: Infected larva dies, releasing new
virus that can infect other larvae
− Vertical: Female that survives a sublethal
infection as a larva can be pass virus on to her
offspring
− Latent infection: Virus lies “dormant” in host,
Fat body infected with CpGV until stress causes outbreak (e.g. overwintering)

6. Symptoms of CpGV Infection (Laboratory)
Healthy CM larva Larva killed by CpGV

7. Symptoms of CpGV Infection (Field)
Failed entry Larva infected Larva probably infected as late
(“sting”) as early instar instar or with low dose

8. Control of Codling Moth Larvae on Delicious Apple (2003)
Investigator: Dr. Helmut Riedl, Oregon State University
Location: Hood River, OR
100
All sprays initiated 250 DD after biofix.
Cyd-X applied weekly, Entrust & Guthion 4 times in 400 gpa.
Four single tree replicates/treatment
CM Injuries per 100 fruit
80
Stings
60 Entries
40
20
0
Untreated CYD-X Entrust Guthion
control 3 oz 2 oz 2 lb
(+ 32 oz
CER-2003-081 NuFilm 17)

9. Infection route & application timing:
Codling • Virus must be ingested by larva to initiate infection.
moth egg
• Larva continues to feed until virus replicates enough
to kill it (can require several days).
• Neonate larvae wander after hatching to seek fruit
(usually within 1-2 days).
• Most don’t eat until they enter fruit (may drink water
droplets and sample foliage).
• Don’t ingest skin when making entry hole.
• Virus picked up on body and mouthparts will
contaminate feeding site & infect larva when it starts
to feed.
• Target hatching eggs and wandering larvae

10. Phenology of the Codling Moth
Source: L. Gut & J. Wise
Michigan State University Extension Bulletin E-154*
CYD-X application periods
Apr May Jun Jul Aug Sep Oct
adults
larvae
Bloom
egg
laying
hatch
0 250 1000 1300 2100
Degree-days (Base 50°F) after Biofix
* Available online at http://www.msue.msu.edu/epubs/pestpubs/E154/18-CodlingMoth.pdf

11. Environmental persistence
• CpGV can persist for years in soil, leaf litter, etc.
• Half-life is approx. 2 days in sunlight
• Longer persistence (7 days +) on shaded foliage
• Common recommendation: reapply after 7-8 sunny days
• Two sprays at ½ rate will give longer protection than 1 spray at full rate
• Spraying late afternoon or early evening better than morning

12. Residual Activity of CYD-X on Apples
Investigator: Dr. L. Lacey, USDA-ARS
Location: Yakima, WA, 2003
Five larvae confined on apples picked 1 to 14 days after application.
3
Untreated check
3 oz/Acre
Live larvae per fruit
6 oz/Acre
2
1
0
0 1 3 7 10 14
Days between application and bioassay of fruit

13. Usage Recommendations
• Application rate
− 1 to 6 fluid ounces per acre (label)
− 3 oz/acre is typical rate
• 2 - 4 applications per larval generation
• Time 1st application at 0-5% egg hatch (200 – 250 DD
after biofix)
• 2nd application 7-14 days later OR at peak egg hatch
• Subsequent applications at 7-14d intervals if needed
• Use sufficient water to attain thorough coverage

14. Usage Recommendations for CYD-X (cont’d)
• Natural UV protection
“Whole” virus product, not artificially “purified”
No chemical additives or residues
• UV protection can be enhanced to some extent
Pinolene-type stickers, Humic acid, Powdered skim milk, Molasses
• Feeding stimulant may help more than UV screen
Sugar or molasses (5 to 7 lb/Acre)
• Compatible with other orchard pest management tactics
Excellent fit with pheromone-based mating disruption
Tank mix with most insecticides/fungicides/acaricides
Do not mix low rates (< 4 oz/Acre) with copper fungicides
Keep pH between 5 and 8

15. Storage Stability of CYD-X
100 • Study conducted by Dr. L.
Lacey of USDA-ARS in
Yakima, WA
% Larval mortality
• Stored at constant
temperature.
• Samples pulled every 3 mo.
50 Storage temperature • Diluted to 2 ppm for bioassay
-4°F (-20°C) against 1st instar CM larvae.
36°F (2°C) • Equivalent to 1 fl. oz. in
77°F (25°C) 16,000 gal water, 40X more
95°F (35°C) dilute than lowest likely rate.
0 • No significant loss if stored at
0 3 6 9 12 or below room temperature
(25°C).
Storage time (months)

16. Storage of CYD-X:
“90 days below 90 degrees”
• No refrigeration required if used
within 3 months.
• Refrigerate any unused product
and use it first next season.
• Freezing won’t hurt the product.
• Certis keeps product refrigerated
until it is sold.

17. CpGV Use Strategy:
Damage Prevention vs.
Population Reduction
• CpGV applied against 1st
generation larvae
• Virus did not eliminate
damage but resulted in
fewer entries
• High level of delayed
mortality (reduction in
subsequent generation) Corrugated
cardboard
Source: “Managing Codling Moth with Nylon net against birds
Granulovirus and Spinosad” by K. Granger, J. Figure: M. Baggiolini
Brunner, & M. Doerr (Washington State Univ.)
Poster presentation, 2003 Wash. State Hort.
Assoc. meeting, Wenatchee, WA
http://entomology.tfrec.wsu.edu/jfbhome/growerarticles/03-WSHAOrganicCMPoster-JFB.pdf

18. Transition to CpGV and Mating Disruption
(Courtesy of M. Andermatt, Andermatt BioControl)
12
damage larvae/tree
10
8
6
4
2
0
1995 1996 1997 1998 1999 2000 2001 Charmillot et
10.5. 27.5. Pasquier, 2002
Isomate-C Plus
attract and kill fenoxycarbe
12.6. 26.6.
MADEX (CpGV)
no attract and kill diflubenzuron
treatements 2.7. 2.8.
8 x 100 ml 3 x 100 ml 1 x 100 ml 1 x 100 ml
phosalone phosmet
28.7. 14.8. chlor-
-- 4 x 50 ml 6 x 50 ml 5 x 50 ml
diflubenzuron pyrifosmethyl

19. CYD-X Program in
1st GV use
Organic Apples (2003)
Source: Jay Brunner & Mike Doerr,
Washington State University
(http://entomology.tfrec.wsu.edu/
jfbhome/growerarticles/)

20. CYD-X Program in Organic Apples (2003)

21. Recommended CYD-X Program in
Conventional Orchard
• 2 to 4 applications against 1st larval generation
• Rotate/mix with other insecticide for more rapid knock-down if desired
– Example: Rimon (novaluron) as ovicide
• Switch to chemical insecticides for 2nd larval generation
• Tank-mix or rotate CYD-X for resistance management
• Population management
− Virus will reduce number of 2nd flight moths produced by 1st brood,
− Control of 2nd generation larvae will be easier
− Virus will shift injury from deep entries to stings
− Sting injury from 1st generation larvae is of lower economic impact
− Rely on chemical insecticides to prevent direct damage by 2nd generation
− Virus will reduce overwintering population

22. Control of Codling Moth Damage on “Spur Rome” Apples (2006)
Investigator: Dr. James Walgenbach, NC State University
Location: Dana, North Carolina
• Commercial orchard block (1.8 acres), 4 reps with 4-5 trees per plot.
• Initial CM application at 250 DD after biofix, except Rimon (100DD as ovicide).
• Airblast sprayer delivering 100 GPA.
Percent codling moth damage
6 September
Treatment* 20 July Stings Entries Live worms
Untreated control (no CM insecticides) 4.5 5.8 10.0 3.0
4 oz Assail 30SG (4X) alt. 16 oz Intrepid 2F (3X) 3.0 5.5 7.3 2.5
5 oz Assail 30SG (4X) alt. 16 oz Intrepid 2F (3X) 2.0 2.5 4.8 0.8
3 oz CYD-X (7X) alt. 5 oz SpinTor 2SC (4X) 1.5 3.3 3.3 1.0
2 lb Guthion 50WP (6X), 16 oz Intrepid 2F (1X) 2.0 2.0 3.3 1.0
20 oz Rimon 0.83EC (4X) alt. 6 oz Calypso 4SC (4X) 1.3 2.5 1.8 1.0
30 oz Rimon 0.83EC (4X) alt. 6 oz Calypso 4SC (4X) 0.3 3.0 0.8 0.5
IND-2006-039

23. Control of Codling Moth in Apples with Mating Disruption & CpGV (2006)
Investigator: Dr. Larry Hull, Penn State Univ. Fruit Research Center
Location: Arendtsville, Pennsylvania
0.5
• 30 acre commercial orchard, 2nd year under
mating disruption and CYD-X.
Isomate/CYD-X 0.5
5 larvae • Isomate CM/OFM (twin tube) at 200/acre.
0.4 Isomate/CYD-X 1.0
Conventional • CYD-X applied at 0.5 or 1.0 oz/acre by ARM (25
GPA/side), + 2 oz. pinolene sticker
% apples with frass
• 5 applications per CM brood (10 total)
0.3
• Conventional program: Rotational program of
Guthion, Imidan, Intrepid (ARM)
• Isomate/Cyd-X program included other
0.2 insecticides for other pests (Assail, Avaunt,
1 larva Provado, etc.)
• Lowest damage and worm populations in block
0.1 that received only 5 oz. total CYD-X
• Infestation decreased in block treated at 1 ounce
0 larvae despite 55% lower application rate than same
block in 2005 (10 oz vs. 22 oz total).
0.0
2-Jul 16-Jul 30-Jul 13-Aug 27-Aug 10-Sep 24-Sep
CER-2006-010