Quarantine and Pre-Shipment. Quarantine and Phytosanitary Treatments. Treatments. Treatments. Quarantine and Pre-Shipment. Methyl Bromide Fumigation

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Quarantine and Phytosanitary s Beth Mitcham Department of Plant Sciences University of California USA Quarantine and Pre-Shipment s Required to prevent spread of economically important insects and diseases Should be scientifically based Not designed to achieve zero risk Generally cause some reduction in product quality from treatment Quarantine and Pre-Shipment s Pests may be internal or external Economic pests on the host or hitchhikiking Can be multiple pests and multiple life stages of each pest to be controlled Methyl Bromide Fumigation Primary fumigant used General biocide Has been inexpensive and easy to use Cost has increased greatly Worker safety issues Phasing out for all uses exempt quarantine Alternatives are under development Current & Potential Methods for Postharvest Insect Control Methyl bromide fumigation Other fumigants Surfactants Heat treatment hot water hot air radio frequency Cold treatment Irradiation Silcone-based Surfactants to Control Surface Pests on Cherries

Issues for California Cherries Variety of insects and mites found on surface of sweet cherries Generally not significant for production Create issues for trade Methyl bromide fumigation for Australian market 5% of fumigated shipments retreated due to finds of live mites Costly Negative effect on fruit quality Potential Surface Arthropods on Fruit Tydeid mites Two-spotted spider mite Western flower thrips Grape mealybug Oblique-banded leaf roller LBAM (potential new pest in Calif.) Scale insects Issues Effort to prove non-host status of sweet cherry for codling moth in Japan Without methyl bromide fumigation, surface pests could become a greater problem for trade Great interest in a reliable method to remove or kill surface pests on sweet cherries Tydeid Mite Two-spotted Spider Mite Western Flower Thrips

Grape Mealybug Obliquebanded Leafroller Mortality of Grape Mealybug Crawlers after with Silwet L77 Mortality of Arthropods after with Silwet L77 % Mortality # Treated Western flower thrips Pacific spider mite Cotton aphid Control 9 control 7 H 7 H 6 8.% 6.7.% 9 96 9.5%.5% 99 99 98 % 95.5% 97 99 Mitcham et al, Mitcham et al, Mortality of eggs of various arthropods after treatment with Silwet L77 control H.% Grape mealybug.7.6. Pacific spider mite 8.5.7 99. Omnivorous leafroller. 5..9 Cherry Project 5 Test fruit tolerance to Silwet L77 and other polydimethyl siloxane surfactants Test removal of and mortality of remaining arthropod pests.5%.6..5%. 99.5.7 Mitcham et al,

Fruit Quality & Pest Tests Evaluation Times Simulated air shipment days at 5C plus 5 hours at C reps of 5 cherries Solution and fruit at C Swirl minute Blot on absorbant towels Store in cherry zip bags Air and Sea Shipment Simulated Sea shipment days at C plus 5 hours at C.5.5 Air Sea Pitting Score.5 Untreated control.5 control H O...5.5.5.5.5 control Defoamer Silwet L-77 SQ Type A Type B 6 SG =slight; = moderate; =severe (%) Fruit quality after treatment and storage % Silwett L-77 Artificial Infestation Two spotted spider mites Removal Mortality Artificial Infestation Cherries gently removed from solution Solution filtered with low suction Count eggs and other life stages removed and remaining on cherries

Water reservoir Perforated plate Water tank Test bed Mesh Pump Y junction Overflow 7 L /second Metal brackets Garden hose Plastic stand Mean % Removal Mite Removal Following with Surfactants 9 8 7 6 5 % mites removed % eggs removed Test bed side and top Shower H O only...5.5.5.5 control Defoamer Silwet L-77 SQ 6 SG Type A Type B (%) Mite Mortality Following with Surfactants 9 Cherry Stem Browning and Surface Pitting after and Simulated Sea Shipment Mean % Mortality 8 7 6 5 Untreated control Water only control.% Agrinse 7 Stem Browning. a. a. a minute Pitting. a. a.9 a Stem Browning.5 cd.5 cd - 5 minute Pitting.5 a.6 a -.6% Agrinse 7. a.7 a.9 a.5 a H O only...5.5.5.5 control Defoamer Silwet L-77 SQ 6 SG Type A Type B.9% Agrinse 7 ppm chlorine. a. a.7 a. a.6 bc.7 ab.6 a.5 a (%) Rating scale: (none), (slight), (moderate), (severe) Survivorship of Two-spotted Spider Mites and Western Flower Thrips after a 5 Minute Shower Removal of Two-spotted Spider Mite Eggs after a or 5 Minute Shower Two-spotted spider mite Mean # survivors Mean # remaining Western flower thrips Mean # survivors Mean # remaining *Untreated control Mean # Eggs Remaining minute 5 minute 76 76 *Untreated control Water only control.% Agrinse 7 7 a 9 a 76. a. a 6. 5.7 Water only control.% Agrinse 7.6% Agrinse 7.9% Agrinse 7 a a -. a 5 a - a 6.7 a.9% Agrinse 7 a 6. a.7 5

Mortality of Two-spotted Spider Mites Eggs after a 5 Minute Shower Untreated control Water only control.6% Agrinse 7.9% Agrinse 7 Mean % Mortality (SE). (.8) 8.5 (.). (.) 6.9 (.) Hot Water In Collaboration with Jim Hansen USDA ARS, Washington State Commercial of Mango Fruit for Quarantine Hot Water Bath System Temperature Profile during Heating of Cherries with 58 C Hot Water for Minutes Temperature ( F) 86 Water Fruit surface Fruit center 68.8.6. Time in hot water bath (min) Hot Water Bath s Temperature Time (min) C Temperature C Time (min) 6 8,,,, 6 5.5,,,, 8 5 5 6, 8, 9,,,, 6, 8,, 5, 6, 7, 8,,,,,, 5, 7, 8, 9 5,,,, 6, 8 56.5,,,, 6 58.5,.5,.75,,, 6

Effects of Hot Water on Attributes of Cherry Quality Fruit Firmness Color Change Soluble Solids Content Titratable Acidity No Significant Effects were Found on these attributes Maximum Time (min) in Hot Water Bath that Resulted in Acceptable Quality Ratings after Simulated Air Shipment Temperature C Berry Browning Stem Color Pitting Overall Acceptability 6 >6 >6 >6 8 > > 9 < 5 7 6 5 >9 7 7 5 > > > 5 >6 56 >6 58 > >/ / Cherries Treated with Hot Water at 5 C C after Simulated Air Shipment Comparison Between Cherries Treated on Harvest Day and One Day Delay Acceptability Index.6... 5ºC, 5min 5ºC, 9min Harvest day One day Delay Untreated Cherry Fruit Tolerance and Codling Moth Mortality Following Hot Water & Simulated Air Shipment 6 Time (min) 8 R =.99 R =.99 Cherry Insect Radio Frequency Heating 6 8 5 5 5 56 58 Temperature ( C) 7

Radio Frequency Heating Polarization of Molecules Polarity is Reversed 7 Million Times per Second Heats much faster than conventional methods Electromagnetic waves interact directly with material to generate heat Preferentially heat insects over commodity Commercially available; used for drying of baked products, textiles, and paper forms On-line systems --- --- H + H + H + H + O - O - +++ +++ RF Generator RF and Microwave Heating Conventional Heating Volumetric heating -- the electromagnetic waves directly couple with material to generate heat. radiation conduction RF or Microwaves moving fluid (air or water) convection h k h = 6 W/m C in still air, W/m C in moving water k ~.5 W/mC RF Heating FCC approved frequencies for ISM Applications:.56 MHz ( m), 7. MHz ( m), and.68 MHz (7. m) Temperature of in-shell walnut kernel when subjected to radio frequency and forced hot air treatments (air temperature, 5 o C; air velocity, m/s) 8

Major Insect Pests in Walnuts Codling moth: field & quarantine Indianmeal moth: storage & phytosanitary Navel orangeworm: : field & phytosanitary Red flour beetle: storage & phytosanitary Thermal Death Kinetics of Four Insect Pests CM = codling moth; IMM = Indianmeal moth; NOW = navel orangeworm; RFB = red flour beetle Source: Johnson et al.,. A 7. MHz, 5 kw RF unit for large-scale tests Diamond of California 5 Radio Frequency for Sweet Cherry Quarantine target: Surface temperature of 6 C C and minimum walnut kernel temperature of 5 C C for 5 min. Shipments to Japan and Australia Methyl bromide will still be allowed after 5 because of official quarantine Methyl bromide causes some stem browning 9

Heating Fresh Fruit with Radio Frequency Energy Requires use of saline solution to prevent burning at contact points Heats very rapidly maybe too rapidly Very low frequency non-thermal pulsed RF may be another option Fiberoptic Probes to Measure Temperatures in the RF Field Minimum Time-Temperature Temperature Combinations for Complete Kill of 5 th Instar NOW, IMM, and CM Time until % mortality (min) 6 Navel orangeworm 5 Codling moth Indianmeal moth 5 5 Temperature ( o F) Temperature Profile during Radiofrequency heating of cherries from RT to 58 C Temperature ( F) 86 Water Fruit surface Fruit flesh middle Fruit center 68.8.6.. RF heating time (min)

Heating Rate of Cherries and Water during Radio Frequency and Hot Water Heating Temperature ( F/min) 5 6 8 Water st min nd min rd min Surface Center RF heating Surface Center Hot water bath Temperature of Cherries and Water after min Heating with Radio Frequency or Hot Water Temperature ( F) 6 9 5 Water Surface Center RF Hot water Heating with RF was.5 C/Min. Faster than in Hot Water Advantages of Radio Frequency Heating Compared With Hot Water Shorter Time Stable and Higher Heating Rate Lower Total Heat Exposure on Fruit Surface where Damage Occurs Should Result in Better Fruit Tolerance Water out Incoming fruit 5 - C RF Heating System Hydro warmer Water heater 5 - F RF heater Water in 5-56 F Fruit to Cooler Temperature ( C) 6 5 Heating rate of cherries Water Fruit RF pretreatment hold drain hot water and add iced water Transfer to C water bath 6 8 Seconds

RF of Sweet Cherries Protocols for Infested Studies Infesting cherry: larva per cherry. Tray with infested cherries (covered with lids). Left overnight before RF treatments Infested fruit chosen for RF treatments. Left: Silk screen bag, cm x.5 cm Center: Cherries inside bag, /bag Right: Tightened bag ready for RF treatment. Larvae inside cherry, left overnight after RF treatment Larvae counted, collected in cups, and later checked for death

Mortality of Codling Moth Larvae after Radio Frequency Heating Effects of RF Heating on Cherry Stem Quality AIR Temp. C 8 5 5 Minutes 6 5 % Mortality.9 89.7 99. Stem Browning browning SEA 5 5.5 6 6 5 6.5.5 Minutes C 7 C 5 C 5.7 C 5. C 5. C Temperature Effect of RF Heating on Cherry Fruit Pitting Effect of RF Heating on Cherry Fruit Firmness AIR AIR Pitting SEA Firmness (g) SEA 6 6 5 6.5.5 Minutes 6 6 5 6.5.5 Minutes C 7 C 5 C 5.7 C 5. C 5. C Temperature C 7 C 5 C 5.7 C 5. C 5. C Temperature Continuous 5 kw RFtube system for treating fresh fruit Hot Water or RF Use very high quality cherries treated with GA Treat on the day of harvest Optimize all handling Air freight to market within a few days

Fruit on or off of the tree are living organisms! Development of Controlled Atmosphere High-Temperature s (CATTS) Lisa Neven and David Obenland USDA ARS, Washington & California Dozing daily Sleeping fruit. Napping nightly. Sleeping fruit. Swaying slightly. Snoozing soundly. Snoring slightly. Sleeping fruit. Fruit on the Tree Nectarine Temperatures on the tree While fruit are on the tree, they experience fluctuating temperatures. In response to these fluctuating temperatures, the fruit produce a number of stress response proteins. The fruit will produce these proteins even if the daily temperatures are not in the stress range every day. :: AM Temperature F 9 8 7 6 :: AM :: AM Time :: AM :: AM C C C C Cherries on the Tree in Tasmania Fruit after Harvest Degrees C 5 5 5 5 5 5 6 7 8 After harvest, the fruit will continue to make the stress response proteins for a number of days. These proteins will help the fruit tolerate heat treatments. -Dec -Dec -Dec 5-Dec 6-Dec 7-Dec 8-Dec 9-Dec -Dec -Dec -Jan Cherry core temperatures in Tasmania, Australia where daily high air temperatures are below C.

Development of the System Heat treatments have been used for disinfesting fresh commodities for thousands of years. Adding a controlled atmosphere to a heat treatment was first investigated in the late 9 s. However, the advent of chemical fumigants stifled any further research in this area. What CATTS Does First, the oxygen level is reduced to a level which will not support active metabolism in the insects. High carbon dioxide levels prevents the insect from down regulating metabolism and respiration. The addition of high temperature causes a metabolic stress in the insect, and large scale systems break down occurs. The addition of CA causes mortality about X as fast as heat alone. Respiratory Response to Heat Fifth instar codling moth CO production during a simulated heat treatment of apple Note characteristic peak followed by rapid decline in CO production. ul CO/mg/min ul CO/mg/min VAPOR 8 C (8 F) 5 5 5 5 5 5 5 TIME (min) MOIST 8 C 5 5 5 5 5 5 5 TIME (min) TEMPERATURE C (8 F) TEMPERATURE C Time CATTS in the Lab Temperature Atmosphere Controlled Atmosphere Temperature System Controls & Monitors: O, CO, Air Speed, Humidity, Dew Point, Air Temperatures, Heat Rate, Fruit Temperatures (surface & core) Commercial CATTS Chamber Placing temperature probes in apples. Technicians after placing a mixed load into the ton CATTS unit. 5

s developed against.. s developed so far Codling moth Oriental fruit moth Plum Curculio Apple Maggot Sweet Cherries Apples Pears, both winter and summer Peaches Nectarines Apricots (in progress) Western Cherry fruit fly Stage of Development Sweet Cherries: Two treatments have been developed for control of codling moth and western cherry fruit fly 5 min at 7 C (7 F) with % O, 5% CO. 5 min at 5 C ( F) with % O, 5% CO. Fruit quality better than MeBr fumigated. No commercial application has met with specifications. Control of bean thrips in navel oranges using ethyl formate fumigation for export to Australia Neven 5. J. Econ. Entomol. 98(): 79-75. Neven & Rehfield-Ray 6. J. Econ. Entomol. 99(): 658-66. Ethyl Formate Natural plant volatile essential oils of grasses, beer, rice, beef, and cheese (Desmarchelier et al., 999). volatile components of grapes and wine (Hiroyasu, 97). Has been widely used as a fumigant for pests associated with dried fruit Registered for use in Australia by BOC gases as Vapormate TM in formulation with CO Previously registered in US for control of insect pests in raisins, but registration expired in the 98 s Results from Table Grape Project Target Pest Life Stage Western Flower Thrips egg Pacific Spider Mite Grape Mealybug Melon Aphid nd instar prepupae adult deutonumph adult adult mixed stages % ethyl formate, hour fumigation at C Simpson et al., 7 Journal Economic Entomology LD 99.99.6.87..9.7.79.6 6

Preliminary Studies Bean Thrips Bean Thrips colony established at UC Davis Mortality studies Field run, unwaxed and unwashed navel oranges Infested with bean thrips Expose to ethyl formate upon arrival Fruit tolerance studies Field run unwaxed, unwashed navel oranges Expose to ethyl formate after one night at 5C and slight warming Laboratory System for Ethyl Formate Fumigation Method Courtesy Joseph Morse and Associates Inlet Port Sample Port Filter Paper Photo courtesy Joe Morse, UCR 7

Mortality of adult bean thrips in the navel of oranges following a -hour exposure to ethyl formate (% ethyl formate)..5.75.5 % Mortality.9a.b.b.b.b Pulp temperature 6 F, air temperature 75 F N 6 7 87 % Ethyl Formate..5.75.5.. Peel Damage Severity following Ethyl Formate Fumigation days days @ 5 C d 5 C + 7d C. (/9).7 (6/9). (6/9).6 (8/9) Damage Scale: = none, = very slight; = slight, = moderate, 5 = severe Damage Scale: = none, = very slight; = slight, = moderate, 5 = severe Fruit Quality after Fumigation with Ethyl Formate weeks storage at F and 7 days at 68F % Ethyl Formate Fruit Quality after Fumigation with Ethyl Formate weeks storage at F and 7 days at 68F % Ethyl Formate No Wax Waxed w/ carnauba No Wax Waxed w/ carnauba Source: Exeter TIs Source: Exeter TIs 8

Mortality of the California Red Scale (Aonidiella aurantii) weeks after a -hour Fumigation with Ethyl Formate EF concentration (%) Instar.75%.5%.% First 99.8 Second 98.8 Third 99.9 Mature third 99.5 Y. Aharoni, Y. Nitzan, and A. Copel. (987) Tropical Science (7) 55-57. 9