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Hot Water Immersion to Ensure Quarantine Security for Cryptophlebia Spp

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Hot Water Immersion to Ensure Quarantine Security for Cryptophlebia Spp Exhibit 4b, #33 STORED-PRODUCT AND QUARANTINE ENTOMOLOGY Hot Water Immersion to Ensure Quarantine Security for Cryptophlebia spp. (Lepidoptera: Tortricidae) in Lychee and Longan Exported from Hawaii PETER A. FOLLETT AND SUZANNE S. SANXTER U.S. PaciÞc Basin Agricultural Research Center, USDAÐARS, P.O. Box 4459, Hilo, HI 96720 J. Econ. Entomol. 94(5): 1292Ð1295 (2001) ABSTRACT We determined whether immersion in 49ЊC water for 20 min, a quarantine treatment developed for disinfestation of fruit ßies in lychee, Litchi chinensis Sonn., and longan, Dimocarpus longan (Lourd.) Steud., exported from Hawaii, would also disinfest fruit of two species of Crypto- phlebia. The pattern of tolerance to heat in Cryptophlebia illepida (Butler) was generally eggs Ͻ neonates Ͻ early instars ϭ lateinstars Ͻ pupae. No C. illepida survived immersion for 16 or 20 min. Late fourth and Þfth instars were determined to be the most tolerant stage that occurs in harvested fruit. Lateinstars of Cryptophlebia ombrodelta (Lower) were more tolerant of hot-water immersion than thoseof C. illepida, but no C. ombrodelta late instars survived immersion for 16 or 20 min. The hot water immersion quarantine treatment for fruit ßies should effectively disinfest lychees and longans of any Cryptophlebia. KEY WORDS Cryptophlebia, quarantine pest, heat treatment, disinfestations HOT WATER IMMERSION at 49ЊC for 20 min is approved by four varieties of longan harvested from orchards in the U.S. Department of Agriculture, Animal Plant Hawaii over a two-year period, Cryptophlebia infes- Health Inspection Service, as a quarantine treatment tation rates were 1.5 and 0.14%, respectively (G. T. for HawaiiÕs tephritid fruit ßy pests attacking lychee, McQuate, USDA, Hilo, HI, personal communication). Litchi chinensis Sonn., and longan, Dimocarpus longan Current regulations for hot-water immersion quar- (Lour.) Steud. (Federal Register 1997). In addition to antine treatment of Hawaii-grown lychee stipulate fruit ßies, two species of Cryptophlebia arealso pests that exported fruits must be found free of Cryptophle- of quarantine concern in sapindaceous fruits (lychee, bia spp. (Federal Register 1997). Thus, the presence longan, and rambutan) exported from Hawaii. Cryp- of Cryptophlebia can prevent export of lychee from tophlebia illepida (Butler), the koa seedworm, is en- Hawaii. The quarantine treatment used for lychee has demic to Hawaii, and C. ombrodelta (Lower), the been proposed for longan also exported from Hawaii litchi fruit moth, is nativeto Australia. Cryptophlebia (P.A.F., unpublished data). If the accepted hot-water spp. are the most commonly intercepted insect pests immersion quarantine treatment for pest fruit ßies is in sapindaceous fruits from Hawaii carried in passen- also effective against C. illepida and C. ombrodelta, gerbaggageand arriving in themail on theU.S. main- these species could be added to the regulation, pre- land (USDAÐAPHISÐPPQ 1997). venting potential interruption of shipments because of Cryptophlebia spp. lay eggs singly on the fruit sur- the presence of these insects. The objectives of the face, and neonates bore through the skin and feed at studies with Cryptophlebia reported here were to ex- the skinÐpulp interface. Although many eggs may be amine the effectiveness of the hot-water immersion laid on thefruit surface,typically only onelarva is treatment against various stages of Cryptophlebia, and found feeding in a fruit. Cryptophlebia spp. arepoten- thereby determine if the approved quarantine treat- tially multivoltine in Hawaii (Jones et al. 1997). Lab- ment provides quarantine security against these pests. oratory tests and Þeld surveys in Hawaii indicate that lycheeandlongan arepoor hosts for Cryptophlebia Materials and Methods (McQuate et al. 2000; P.A.F., unpublished data). Ac- tual damage from larval feeding is often minimal, and Insect Rearing. A colony of C. illepida was started larval survival to pupation is rarewhenfruit areon the using larvae( Ϸ1,000) collected from macadamia nuts tree. However, when a neonate enters the fruit at the from an orchard in Pahala, HI. Because of difÞculties calyx or stem end, it may bore through woody tissue collecting C. ombrodelta in theÞeld, C. ombrodelta to the seed and feed there, improving the chance for pupae(108) wereobtainedfrom a laboratory colony survival to pupation. Successful larval development in maintained at the Maroochy Experiment Station in fruit also may improveoncefruit areharvested Queensland, Australia, to start a colony; the Maroochy (P.A.F., unpublished data). In a study of Ͼ36,000 ma- colony had been maintained in the laboratory for 3 yr ture fruits of six varieties of lychee and 9,700 fruit of and wild moths from infested macadamia were intro- October 2001 FOLLETT AND SANXTER:HOT WATER IMMERSION TO CONTROL Cryptophlebia 1293 caps had TeßonÐsilicone septa to permit insertion of temperature probes. All hot-water immersion treat- ments were done in a 75-liter circulating bath heated by two electric heaters (immersion circulator model 73, PolyScience, Preston Industries, Niles, IL) to 49 Ϯ 0.2ЊC. Bath temperature was veriÞed before and after each run using a mercury thermometer with 0.1ЊC gradations. During treatment, temperature was mon- itored in eight vials at the surface and center of the diet,and at theinsidewall of theglass vial abovethe diet, at 30-s intervals using temperature probes and a data logger (model 5160-64-A, Omni International, Logan, UT) calibrated to a mercury thermometer. Fig. 1. Temperature proÞles of diet in vials and of lychee Vials with individual larvaeor pairs of pupaewere Њ fruit during hot water immersion at 49 C for 20 min followed placed in wire baskets (1 cm mesh) and submersed to by immersion in ambient temperature water for 20 min. the bottom of the tank for the desired treatment time. For larval treatment, baskets with vials were tapped on duced annually to maintain genetic variation. For both a tableto dislodgeinsectsfrom thewall of theglass vial species, adults were placed in 3.8-liter plastic jars with onto the diet surface before submersion. For egg treat- ventilated lids for mating and provided with a 5% ment, gravid females were conÞned to 29-ml plastic honey water solution through a wick. Eggs were laid cups overnight, and open cups with eggs were placed on theinsidesurfaceof thejar. Emergingneonates in wire cages and treated as above (i.e., eggs were in were transferred daily to 29-ml diet cups with labo- direct contact with the water.) Regardless of treat- ratory diet. The laboratory diet was modiÞed from ment time, the basket with vials or cups and insects Sinclair (1974) as described in Follett and Lower was transferred to a 75-liter ambient-temperature Ϸ Њ (2000). As larvae approached pupation, diet cups ( 22Ð24 C) cooling bath for 20 min after heat treat- (with larvae) with lids removed were placed in 3.8- ment. (Although not part of the ofÞcial treatment, a liter plastic tubs (Rubbermaid, Wooster, OH) with 20-min cooling bath is recommended after hot water 30 g of sand covering the bottoms to a depth of 0.25 cm. immersion of lychee fruits.) Larval mortality was mea- C. illepida larvaetypically leavethediettrays to pu- sured after 24 h by prodding larvae with a camels-hair patein thesand, whereas C. ombrodelta typically pu- brush for signs of any movement. Pupae were held pate in the diet. Emerging moths were cooled for after treatment in 29-ml plastic cups at 25ЊCfor2wk handling and transferred to the plastic mating jars. for adult emergence. Treated eggs attached to plastic Rearing conditions were 25 Ϯ 2ЊC and a photoperiod cups were held for 1 wk at 25ЊC for neonate emer- of 10:14 (L:D) h for the duration of the experiments. gence. For stage-speciÞc tests, larvae were placed into cate- The two-year survey mentioned earlier that exam- gories (L1, L2/3, and L4/5 corresponding to instars) ined the fruit pests in lychee and longan in Hawaii based on size and known developmental rates (un- showed mean fruit weights of 17.2 g (SD ϭ 4.8, n ϭ published data). 36,351) and 9.4 g (SD ϭ 2.3, n ϭ 9,700), respectively Hot Water Immersion Treatment. Cryptophlebia (G. T. McQuate, personal communication). In pre- are difÞcult to rear on fresh lychee and longan; there- liminary studies, the heating rate at the seed surface of fore, all heat treatments were applied to Cryptophlebia lycheefruitof typical size(17Ð19 g) rangedfrom 3.1 Ϫ in 29-ml plastic cups (eggs) or 25-ml glass vials con- to 3.3ЊC min 1 during theÞrst 7 min of thetreatment taining 10 g of artiÞcial diet (larvae and pupae). Direct (thetimeperiodwhentherateof heatingis thegreat- exposure to hot water of eggs attached to the inner est). Diet was heated so that the thermal proÞle at the surfaceof cups is similar to exposureonthesurfaceof surfaceof thedietin vials mimickedtheproÞlefor the fruit. The amount of diet required in vials was deter- seed surface of a typical lychee fruit (Fig. 1). The mined by comparing thermal diffusion proÞles of dif- heating rate at the diet surface in our tests averaged ferent amounts of diet with that of lychee fruit. Vial 3.3ЊC minϪ1 during the Þrst 7 min after immersion. The Table 1. Mean ؎ SEM percentage mortality of C. illepida after immersion in water at 49°C a Immersion Mortality time, min Eggs (1 day old) Eggs (3 d old) L1 L2/3 L4/5 Pupae 0 586 13.7 Ϯ 3.8 231 19.8 Ϯ 9.1 97 13.8 Ϯ 5.2 94 0.0 Ϯ 0.0 111 1.0 Ϯ 1.0 118 5.5 Ϯ 3.1 4 585 100.0 Ϯ 0.0 569 100.0 Ϯ 0.0 157 68.2 Ϯ 4.8 116 16.2 Ϯ 5.7 123 4.2 Ϯ 1.8 162 17.4 Ϯ 2.3 8 624 100.0 Ϯ 0.0 466 100.0 Ϯ 0.0 152 97.4 Ϯ 2.0 114 51.4 Ϯ 12.5 124 35.6 Ϯ 6.2 158 16.1 Ϯ 4.5 12 686 100.0 Ϯ 0.0 452 100.0 Ϯ 0.0 137 100.0 Ϯ 0.0 115 96.7 Ϯ 3.3 121 98.3 Ϯ 1.7 160 95.3 Ϯ 3.0 16 522 100.0 Ϯ 0.0 426 100.0 Ϯ 0.0 121 100.0 Ϯ 0.0 101 100.0 Ϯ 0.0 106 100.0 Ϯ 0.0 155 100.0 Ϯ 0.0 20 698 100.0 Ϯ 0.0 378 100.0 Ϯ 0.0 114 100.0 Ϯ 0.0 78 100.0 Ϯ 0.0 99 100.0 Ϯ 0.0 149 100.0 Ϯ 0.0 a Mortality of each life stage is preceded by the number of test subjects.
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