Insecticidal virus

for control of

Codling Moth larvae

Active ingredient:

Codling moth granulosis virus

• Cydia pomonella granulovirus (CpGV)

• Natural pathogen of codling mothOcclusion

body (OB)

Nucleocapsid

Granulin

protein

Glycoprotein

envelope

Viral

DNA

• 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

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

CpGV Products Registered with the

US EPA (2005)

*OB = occlusion bodies

• 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!)

• 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,

until stress causes outbreak (e.g. overwintering)

Biology & Mode of Action of CpGV

Fat body of healthy larva

Fat body infected with CpGV

Healthy CM larva Larva killed by CpGV

Symptoms of CpGV Infection (Laboratory)

Larva probably infected as late

instar or with low dose

Failed entry

(“sting”)

Larva infected

as early instar

Symptoms of CpGV Infection (Field)

Control of Codling Moth Larvae on Delicious Apple (2003)Investigator: Dr. Helmut Riedl, Oregon State University

Location: Hood River, OR

0

20

40

60

80

100

Untreated

control

CYD-X

3 oz

Entrust

2 oz

Guthion

2 lb

CM

Inju

ries p

er

100 f

ruit

Stings

Entries

All sprays initiated 250 DD after biofix.

Cyd-X applied weekly, Entrust & Guthion 4 times in 400 gpa.

Four single tree replicates/treatment

(+ 32 oz

NuFilm 17)CER-2003-081

• Virus must be ingested by larva to initiate infection.

• 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

Infection route & application timing:

Codling

moth egg

Phenology of the Codling MothSource: L. Gut & J. Wise

Michigan State University Extension Bulletin E-154*

* Available online at http://www.msue.msu.edu/epubs/pestpubs/E154/18-CodlingMoth.pdf

Apr May Jun Jul Aug Sep Oct

0 250 1000 1300 2100

Degree-days (Base 50°F) after Biofix

Blo

om

egg

layinghatch

larvaeadults

CYD-X application periods

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

0

1

2

3

0 1 3 7 10 14

Untreated check

3 oz/Acre

6 oz/Acre

Days between application and bioassay of fruit

Liv

e la

rvae p

er

fruit

Residual Activity of CYD-X on ApplesInvestigator: Dr. L. Lacey, USDA-ARS

Location: Yakima, WA, 2003

Five larvae confined on apples picked 1 to 14 days after application.

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

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

Storage Stability of CYD-X

• Study conducted by Dr. L.

Lacey of USDA-ARS in

Yakima, WA

• Stored at constant

temperature.

• Samples pulled every 3 mo.

• Diluted to 2 ppm for bioassay

against 1st instar CM larvae.

• Equivalent to 1 fl. oz. in

16,000 gal water, 40X more

dilute than lowest likely rate.

• No significant loss if stored at

or below room temperature

(25°C).

% L

arv

al m

ort

alit

y

0

50

100

0 3 6 9 12

Storage time (months)

Storage temperature

-4°F (-20°C)

36°F (2°C)

77°F (25°C)

95°F (35°C)

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.

Source: “Managing Codling Moth with

Granulovirus and Spinosad” by K. Granger, J.

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

CpGV Use Strategy:

Damage Prevention vs.

Population Reduction

• Virus did not eliminate

damage but resulted in

fewer entries

• High level of delayed

mortality (reduction in

subsequent generation)

• CpGV applied against 1st

generation larvae

Corrugated

cardboard

Nylon net against birds

Figure: M. Baggiolini

0

2

4

6

8

10

12

damage larvae/tree

Charmillot et Pasquier, 2002

Transition to CpGV and Mating Disruption(Courtesy of M. Andermatt, Andermatt BioControl)

1995 1996 1997 1998 1999 2000 2001

10.5.

attract and kill

27.5.

fenoxycarbe

no

12.6.

attract and kill

26.6.

diflubenzuron

treatements 2.7.

phosalone

2.8.

phosmet8 x 100 ml 3 x 100 ml 1 x 100 ml 1 x 100 ml

28.7.

diflubenzuron

14.8. chlor-

pyrifosmethyl-- 4 x 50 ml 6 x 50 ml 5 x 50 ml

Isomate-C Plus

MADEX (CpGV)

CYD-X Program in

Organic Apples (2003)1st GV use

Source: Jay Brunner & Mike Doerr,

Washington State University

(http://entomology.tfrec.wsu.edu/

jfbhome/growerarticles/)

CYD-X Program in Organic Apples (2003)

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

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

Treatment*

6 September

20 July

Percent codling moth damage

Control of Codling Moth Damage on “Spur Rome” Apples (2006)Investigator: Dr. James Walgenbach, NC State University

Location: Dana, North Carolina

IND-2006-039

• 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.

0.0

0.1

0.2

0.3

0.4

0.5

2-Jul 16-Jul 30-Jul 13-Aug 27-Aug 10-Sep 24-Sep

% a

pp

les w

ith

fra

ss

Isomate/CYD-X 0.5

Isomate/CYD-X 1.0

Conventional

5 larvae

1 larva

0 larvae

• 30 acre commercial orchard, 2nd year under

mating disruption and CYD-X.

• Isomate CM/OFM (twin tube) at 200/acre.

• CYD-X applied at 0.5 or 1.0 oz/acre by ARM (25

GPA/side), + 2 oz. pinolene sticker

• 5 applications per CM brood (10 total)

• Conventional program: Rotational program of

Guthion, Imidan, Intrepid (ARM)

• Isomate/Cyd-X program included other

insecticides for other pests (Assail, Avaunt,

Provado, etc.)

• Lowest damage and worm populations in block

that received only 5 oz. total CYD-X

• Infestation decreased in block treated at 1 ounce

despite 55% lower application rate than same

block in 2005 (10 oz vs. 22 oz total).

Control of Codling Moth in Apples with Mating Disruption & CpGV (2006)Investigator: Dr. Larry Hull, Penn State Univ. Fruit Research Center

Location: Arendtsville, Pennsylvania

CER-2006-010