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Evaluation of Controlled Atmosphere Anoxia Treatments as a Potential Disinfestation Technique for Thrips and Spider Mites in Greenhouses. 1996 proposal

Evaluation of Controlled Atmosphere Anoxia Treatments

as a Potential Disinfection Technique

for Thrips and Spider Mites in Greenhouses

Daniel A. Potter

Executive Summary

Controlled atmosphere (CA) Technology is a standard technique for the post
harvest

handling of fruits, vegetables and stored grains and is one of the main
reasons that high quality

produce is available throughout the year in the U.S. CA technology is a
simple and

environmentally safe way to manage post harvest insect and disease problems
in cold storage.

The use of low oxygen (anoxia) and high carbon dioxide and nitrogen environments
simply

smother insects. Anoxic environments could be a practical, cost-effective
and safe way to kill all

life stages of insects and mites that infest greenhouse flower crops. Unfortunately,
nearly all

information on effects of anoxic environments on insects concerns pests
of fruits, vegetables and

grains in storage.

This proposal evaluates the potential for using CA anoxia treatments to
control thrips and

two-spotted spider mites on garden impatiens and New Guinea impatiens,
common and

relatively fragile bedding plants. Combinations of N2, CO2 and O2 concentrations
and treatment

times will be evaluated for their impact on all life stages of thrips and
mites at cold, cool and

warm temperatures. This will be followed by an evaluation of the tolerance
of impatiens to

critical thresholds required for elimination of pests. Finally, impatiens
will be infested with

thrips and mites to evaluate the overall efficacy of CA anoxia treatments
under simulated

production conditions. If successful, it is feasible that truckloads of
bedding plants can be

disinfected with CA as they travel to their destinations or simply overnight
before shipment.

Introduction & Literature Review

The production of flowering plants, usually in protected cultivation, is
an economically

important part of agriculture and the fastest growing sector of agriculture
in grower cash receipts

(Johnson and Johnson 1993). Insects and mites account for significant economic
losses and

large scale pesticide usage in flowering plant production.

Thrips are among the most injurious and difficult to control of the greenhouse
pests.

Thrips not only cause damage by feeding but can also vector the important
pathogen, impatiens

necrotic spot virus, INSV (Lewis 1973, Moorman 1994). Similarly, spider
mites are an

important part of the greenhouse pest complex. Spider mites cause damage
by piercing plant

cells and feeding on the contents and by producing webbing on the host
plant. Consequences of

spider mite feeding for host plants include reduced vigor in flowering,
growth and yield

(Tomczyk and Kropczynska 1995). The webbing itself can reduce the marketability
of any

floricultural crop (Van De Vrie 1985).

Cultural, biological and chemical methods are used to control greenhouse
pests but all

have disadvantages. Cultural control practices for thrips and spider mites
include exclusion by
MA

screening, beginning the production cycle with pest-free propagules, and
maintaining a pest-free

facility (Baker 1994). Predaceous mites and thrips, as well as fungal pathogens,
are sometimes

effective for biological control of spider mites and thrips (Van Lenteren
and Woets 1988,

Lindquist 1995). Chemical controls are generally effective but have the
practical disadvantage

of the restrictions specified in the Worker Protection Standard (WPS).
In addition to these

restrictions, chemical controls can create resistance problems (Van Lenteren
and Woets 1999,

Immaraju et al 1992, Brodsgaard 1994).

Controlled atmosphere (CA) or modified atmosphere technology has been used
by the

gram, fruit, and vegetable industries for disinfection of their products
for at least 20 years.

Factors that can influence the effectiveness of the modified atmosphere
conditions include:

atmospheric composition (percent of carbon dioxide, oxygen, and nitrogen);
relative humidity

(water loss); time; pest species; and pest life stage (Ke and Kader 1992,
Fleurat-Lessard 1990).

The general effect of controlled atmosphere conditions is to remove or
limit the oxygen source of

arthropod pests. Typically, as the oxygen concentration decreases, the
rate of kill increases (Ke

and Kader 1992, Fleurat-Lessard 1990). Nitrogen gas alone induces hyperactivity
followed by

immobilization as a result of the anoxia or hypoxia. Carbon dioxide, like
nitrogen, causes

immobilization but it also induces opening of the spiracles. Thus, one
of the primary effects of

carbon dioxide is the loss of water by insects exposed to anoxic conditions
which ultimately

leads to mortality. Carbon dioxide alone at concentrations of 35-60% gives
almost 100% control

of all life stages of most stored grain pests (Fleurat-Lessard 1990).

Mites are generally harder to kill than insects. Age, condition (active
or quiescent) and

feeding status of arthropods can also influence their tolerance to elevated
levels of carbon

dioxide, or hypercarbic conditions. Hypercarbic conditions are also linked
to physiological

disorders such as decreased fecundity or longevity as well as disrupted
metamorphosis (Fleurat-

Lessard 1990).

In addition to the effect of CA on pests, the effect on the crop plant
is also important

Anoxic conditions have been used to disinfect cutflowers during storage
and/or transport to

market (Zheng et al 1993). Most of the insecticidal CA research has been
done at temperatures

in the range of 0-12C (Aharoni et al 1981, Bond and Herne 1983, Zheng et
al. 1993), while

other work (Akluting et al. 1991, 1995) has been at temperature ranges
of 20-40′C. Cutflowers

and chrysanthemum cuttings can be treated at temperatures of 0-10′C, but
potted plants and

Potter and Anderson - Anoxia Effects r,9 Greenhouse Insects

some other propagules cannot tolerate temperatures that low without suffering
quality loss

(Kooten and Peppelenbos 1993, Zheng et al. 1993).

Presently, most CA research for controlling Western flower thrips and spider
mites is

being done on fruit and vegetable crops. Aharoni et al (1981) demonstrated
that high levels of

carbon dioxide (50-90%) or low levels of oxygen (<1%) were successful
for killing thrips on

strawberries. For 100% control, hypercarbic conditions (90%) were required
for 48 h. Zheng et

al (1993) tested longer storage periods and found that low carbon dioxide
(10%) killed more

effectively in 6 days at lower temperatures than decreased oxygen for 2-3
weeks. Spider mite

eggs on apples in cold storage were killed in 4 days at 12C with both a
75% carbon dioxide and

1% oxygen treatment and a 9% carbon dioxide and 0.5% oxygen treatment (Bond
and Herne

1983). Whiting and van den Heuval (1995) killed mite eggs with 113 h exposure
to 0.4%

oxygen and 20% carbon dioxide.

Objectives and Anticipated Benefits

This project involves the development of a simple and safe technique fbr
bedding plant

growers to remove thrips and mites from their plants. It is primarily directed
at the relatively

large cutting and plug producers who inadvertantly distribute insect pests,
especially thrips, to

the hundreds of small and medium size greenhouses across the U.S. Although
the project is

primarily directed at large growers, the potential benefit to small growers
is immeasurable.

From our extension experience in Kentucky, most insect problem seen by
smaller growers come

from some of the plant stock they have purchased. Large growers make many
efforts to prevent

the shipment of insect pests, but it is difficult to completely disinfect
plant material with

conventional insecticides because of incomplete spray coverage, presence
of relatively

nonsusceptible life stages (e.g., eggs) or pest resistance. If successful,
the proposed research

might be applied in the following scenario:

A plug grower collects plants for tomorrow’s shipment. Plants are boxed
in the

afternoon and placed into a sealed fumigation chamber in the shipping area.
A

slight vacuum is created in the chamber and a mixture of nitrogen and carbon

dioxide gases is released, sufficient to replace 99.9% of the oxygen in
the

chamber. After overnight treatment, the N2 and CO2 are released into the

atmosphere and the low oxygen conditions have safely killed all the life
stages of

all the insect and mite pests on the plants.

Before CA technology can be adapted for greenhouse pest management, the
effects of

low oxygen (anoxia) enviromnents on thrips and mites must be determined.
The proposed

research differs from previous work on post harvest treatments of fruits
and vegetables because

it deals with pests and temperatures relevant to greenhouse production
systems, and because it

considers effects of CA on plant growth and quality as well as on pests.
The first objective will

be to evaluate the impact of anoxia and hypercarbic and hypernitrogen environments
on survival

and reproductive performance of thrips and mites. The purpose is to establish
critical thresholds

for mortality of the various pest life stages in a series of different
gaseous environments.

Second, seed impatiens and New Guinea impatiens will be treated with the
same environments to

determine their compatibility with production of high-quality bedding plants.
Finally, infested

impatiens will receive the best combination of treatments to determine
the overall efficiency of

the system for disinfecting whole plants under realistic production conditions.

Materials and Methods

Insect preparation and rearing. Stock colonies of thrips and mites will
be reared on bean plants

in the insectary. Leaf discs will be cut with a cork borer and placed on
a moistened cotton pad in

petri dishes. Adults will be placed on the leaf disc for 24 hours to allow
oviposition. After that

time they will be removed allowing the eggs to remain. Each life stage
(eggs, larvae, nymphs,

adults) will be tested independently. Active life stages will be obtained
by allowing the eggs to

develop under suitable conditions in the growth chamber. Leaf discs with
eggs or active stages

of thrips or mites will be placed into treatment chambers as described
below.


Plantproduction.
Garden impatiens from seed and New Guinea impatiens
from cuttings will be

used in these experiments because both are very susceptible to thrips and
mites and both are

relatively fragile plants that may react negatively to CA environments.
Impatiens seed and

unrooted cuttings will be purchased from commercial sources. Seed germination
and cutting

development will occur in thrips-free double poly greenhouses equipped
with a Biotherm bench

heating system and a misting/watering boom (East Coast Designs, Inc.).
Plants used in

experiments will be moved to treatment chambers in the laboratory during
exposure or

disinfection treatments.

Treatment chambers. A constant flow system will be used for the
chambers. The gas mixture, at

controlled concentrations, will flow through the testing chambers for the
entire testing period at a

set flow rate in air exchanges per hour. This basic system has recently
given encouraging results

for control of whitefly on poinsettias (Han 1995). Our system was designed
with the assistance

of Dr. Richard Gates (Department of Agricultural and BioSystems Engineering)
and Dr. John

Loughrin (a volatiles expert in the Department of Entomology). The gases
will pass through a

series of filters, moisture traps, flow meters and regulators before flowing
into the chambers.

The gases move into a manifold, designed to ensure equal resistance and
distance from the gas

source, where they will mix and produce the desired test mixture needed.
The eighteen

individual chambers, Scienceware’ chemical-resistant vacuum desiccators
(10L volume), will

have a septum for sampling the atmosphere during each experiment. Samples
obtained through

the septa will be analyzed on a head space analyzer gas chromatograph.

CA environments. Temperature, time of exposure and the mixture of
gases are the important

variables for this research. Temperatures of 10, 20 and 30C will be used
while the treatment

chambers are maintained inside a Percival growth chamber. Exposure times
of 0, 12, 24 and 48

h will be used initially to determine overall effects of the CA treatments
and refined in later

experiments. The gases used in the experiments will be oxygen, nitrogen,
and carbon dioxide.

All gases will be obtained as bottled gas from a local source (Scott Gross,
Inc.; Lexington, KY.).

The nitrogen will be Ultra High Pure (UHP) grade similar to that used in
other studies (Whiting

et al. 1991, Whiting and Heuvel 1995). Carbon dioxide purity will be better
than food grade due

to extra container cleaning during processing. Medical grade oxygen will
be used to provide

Potter and Henderson - Anoxia Effects on Greenhouse Insects

precise control of oxygen levels. Gas mixtures with 0 or 5% O2, 0, 5, 10,
15 or 20% CO2 and 80,

85, 90, 95 or 100% N2 will be used in treatments with different exposure
times and different

exposure temperatures. Each CA environment will be replicated six times.
Initially, separate

experiments will conducted an insects or mites on leaf discs and on non-infested
impatiens

plants to determine the range of effects of the CA environments. Mortality
of pests will be

evaluated by direct inspection. Following exposure, impatiens plants will
be grown under

normal production conditions and evaluated for growth effects and days
to flowering. Later

experiments will utilize impatiens plants infested with thrips and mites
to determine specific gas

mixtures, treatment temperatures and exposure time for the optimum control
of the thrips and

mites.

Literature Cited

Aharoni, Y., J.K. Stewart and D.G. Guadagni. 1981. Modified atmospheres
to control Western

Flower Thrips on harvested strawberries. J. Econ. Entomol. 74: 338-340.

Baker, J.R. 1994. Insect and mites. pp.257-271. In E.J. Holcomb[ed.] Bedding
Plants IV. Ball,

Batavia, Illinois.

Bond, EJ. and D.H.C. Heme. 1983. The potential of carbon dioxide and low-oxygen
atmospheres

for control of winter eggs of the European Red Mite, Panonychus ulmi (Acari:

Tatranychidae), on harvested apples. Proc. Entomol. Soc. Ont. 114:11-14.

Brodsgaard, H.F. 1994. Insecticide resistance in European and African strains
of Western flower

thrips (Thysanoptera:Thripidae) tested in a new residue-on-glass test.
J. Econ. Entomol.

87:1141-1146.

Fleurat-Lessard, F. 1990. Effects of modified atmospheres on insects and
mites infesting stored

products, pp.21-38. In M. Calderon and R. Barakai-Golan[eds.], Food Preservation
by

modified atmospheres. CRC Press, Boca Raton, FL.

Han, S. 1995. Personal telephone interview.

Immaraju, J.A., T.D. Paine, J.A. Bethke, K.L. Robb and J.P. Newman. 1992.
Western flower

thrips (Thysanoptera:Thripidae) resistance to insecticides in coastal California
greenhouses. J.

Econ. Entomol. 85:9-14.

Johnson, D.C. and Johnson, T.M. Financial Performance of U.S. Floriculture
and Environmental

Horticulture Farm Businesses, 1987-1991. U.S. Dep. Agric. Statistical Bull.
N2.

Ke, D. and A.A. Kader. 1992. Potential of controlled atmospheres for the
postharvest insect

disinfection of fruits and vegetables. Postharv. News Info. 3:31N-37N.

Kooten, O.V. and H. Peppelenbos. 1993. Predicting the potential to form
roots on Chrysanthemum

cuttings during storage in controlled atmospheres. pp.610-619. In Proceedings,
6th

International Controlled Atmosphere Research Conference, 15-17 June 1993.
Ithaca, NY.

Lewis T. 1973. Thrips: their biology, ecology and economic importance.
Academic, New York,

NY.

Lindquist, R.K. 1995. Where we stand on western flower thrips. GMPro -
Greenhouse Management

Potter and Anderson - Anoxia Effects on Greenhouse Insects

and Production. March pp. 84-88.

Moorman, G.W. 1994. Bedding plant diseases. pp.245-255. In E.J. Holcomb
[ed.]. Bedding Plants

IV. Ball, Batavia, Illinois.

Tomczyk, A. and D. Kropczynska. 1985. Effects on the host plant. pp.317-329.
In W. Helle and

M.W. Sabelis[eds.], Spider mites their biology, natural enemies and control.
vol 1A.

Elsevier, New York, NY.

Van De Vrie, M. 1985. Control of Tetranychidae in crops: Greenhouse ornamentals.
pp. 2773-283.

In W. Helle and M.W. Sabelis[eds.], Spider mites their biology, natural
enemies and control.

vol 1B. Elsevier, New York, NY.

Van Lenteren, J.C. and J. Woets. 1988. Biological and integrated pest control
in greenhouses.

Annu. Rev. Entomol. 33: 239-269.

Whiting, D.C., S.P. Fester & J.H. Maindonald. 1991. Effect of oxygen,
carbon dioxide, and

temperature on the mortality responses of Epiphyas pottsvittanana (Lepidoptera:
Tortricidae). J.

Econ. Entomol. 84: 1544-1549.

Whiting, D.C. and J. van den Heuvel. 1995. Oxygen, carbon dioxide, and
temperature effects on

mortality responses of diapausing Terranychus urricae (Acari: Tetranychidae).
J. Econ-

Entomol. 88: 331-336.

Zheng, J., M.S. Reid, D. Ke, and M.I. Cantwell. 1993. Atmosphere modification
for postharvest

control on thrips and aphids on flowers and green leafy vegetables. pp.
394-401. In

Proceedings, 6th International Controlled Atmosphere Research Conference,
15-17 June 1993.

Ithaca, NY.

Budget requirements for controlled atmosphere anoxia research

Supplies

Pipe, fittings, regulators, flow meters, chambers for
3000.

control of 3 gases

N2, CO2 and O2 gases
500.

Use of greenhouse and growth chamber space
1500.

Plants, growing media, containers
2500.

Total
7500.

Personnel

Principal Investigator: Daniel A. Potter, Professor, Department
of Entomology, 17 yrs

research experience with urban landscape and greenhouse IPM, insect/plant
interactions,

more than 90 refereed articles. 14 book chapters. Education: B.S. degree,
Cornell

University, 1974; Ph.D, Ohio State University, 1978 (dissertation topic
dealt with biology of

the twospotted spider mite). Honors: Distinguished Achievement Award in
Urban

Entomology, Entomological Society of America, 1995; T.P. Cooper Award for
Outstanding

Research Program, Univ. KY College of Agriculture, 1989; about 30 invited
symposium

presentations worldwide.

Selected Publications Originating from Potter’s Laboratory:

Potter, D.A., D.L. Wrensch and D.G. Johnston. 1976. Aggression and mating
success in

male twospotted spider mites. Science 193: 160-161.

Potter, D.A. and R.G. Anderson. 1982. Resistance of ivy geraniums to the
twospotted

spider mite. J. Amer. Soc. Hort. Sci. 107-1089-1092.

Potter, D.A. and S.K. Braman. 1991. Ecology and management of turfgrass
insects.

Annual Review of Entomology 36: 383-406. (Invited)

Potter, D.A. and P.G. Spicer. 1993. Seasonal phenology, management and
host preferences

of the potato leafhopper on nursery-grown maples. J. Environ. Hort. II:
101-106.

Spicer, P.G., D. A. Potter & R.G. McNiel. 1995. Resistance of flowering
crabapple

cultivars to defoliation by the Japanese beetle. J. Econ. Entomol. 88:
979-985.

Loughrin, J.H., D.A. Potter & T.R. Hamilton-Kemp. 1995. Volatile compounds
induced by

herbivory act as aggregation kairomones for the Japanese beetle (Popillia
japonica Newman).

J. Chem. Ecol. 21: 1457-1467.

Principal Investigator: Robert G. Anderson, Professor of Extension Floriculture;
B.S. 1970,

Univ. of Minnesota; M.S. 1973, Univ. of Minnesota; PhD 1976, Univ. of Florida;
has

expertise in cut flower and bedding plant production and marketing.

Selected publications and presentations:

R.G. Anderson. Production of Field Grown Cutflowers. New Crops Symposium.
Indiana

Horticultural Congress, Indianapolis, Jan. 1996.

R.G. Anderson and J. Browne. Single stem cut flower production of corn
flower. Third

International New Crops Symposium, Indianapolis IN, Oct. 1995.

Chao, K., R.S. Gates, RG. Anderson. 1994. Neural-fuzzy inference system
for daily growth

of single stem roses. ASAE Paper No. 944015.