Cold treatment of Ceratitis capitata (Diptera: Tephritidae) in oranges using a larval endpoint

South Africa currently exports fresh citrus (Citrus spp.) fruit to Japan using an in-transit cold treatment protocol of 14 d or 12 d at temperatures <0 degrees C for treatment of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) in 'Clementine' mandarins (Citrus reticulata Blanco) and other citrus types, respectively. To reduce the risk of chilling injury with this treatment, research was conducted with temperatures >0 degrees C. Earlier South African research had shown that young (6-d-old) larvae were slightly more tolerant of cold treatment and that there were no significant differences between cold tolerance of these larvae in different citrus types [oranges, Citrus sinensis (L.) Osbeck; grapefruits, Citrus paradisi Macfad.; lemons, Citrus limon (L.) Burm.f.; and mandarins). Due to their ready availability, 'Valencia' oranges were used in this study. When 62,492 larvae in total were treated in three replicates at a mean temperature of 1.5 degrees C for 16 d, there were three larval survivors. The trial was therefore repeated with oranges using a 16-d period at a mean temperature of 1.0 degrees C and a mean of 1.4 degrees C for the hourly maximum probe readings. Three replicates were again conducted and the resultant mean mortality in the control was 8.1% of 21,801 larvae, whereas the cold treatment mortality was 100% of 71,756 larvae. This treatment at a mean temperature of 1 degree C exceeded the Japanese confidence level requirement and also exceeded the Probit-9 mortality level, but not at a confidence level of 95%. These data support the establishment of a treatment protocol of 16 d at temperatures <1.4 degrees C, commencing once all fruit pulp probes reach a temperature of 1 degree C or lower.     

Comparison of in vitro heat and cold tolerances of the new invasive species Bactrocera invadens (Diptera: Tephritidae) with three known tephritids

Bactrocera invadens Drew, Tsuruta & White (Diptera: Tephritidae) is spreading throughout central Africa attacking a variety of fruit; quarantines are placed on fruit from this region that are considered hosts. The only phytosanitary treatment that is commercially available is an ionizing irradiation treatment for all Tephritidae at 150 Gy. The development of other treatments, such as heat, cold, or fumigation, usually requires testing tens of thousands of insects at a dose that provides efficacy and may take several years. It may be possible to shorten the time required to develop treatments by comparing tolerance of a new quarantine pest to tolerances of pests with similar behaviors and modes of infestation for which treatment schedules are available. Cold and heat tolerance ofB. invadens was compared with tolerance of Anastrepha ludens (Loew), Bactrocera dorsalis (Hendel), and Ceratitis capitata (Wiedemann) in vitro. Third-instar B. invadens was no more cold tolerant than the other species when treated in diet at 0.94 +/- 0.65 degrees C and no more heat tolerant than C. capitata when immersed in vials in water at 44.7 +/- 0.1 degrees C. The data at 0.94 +/- 0.65 degrees C was used to include B. invadens in a USDA cold treatment schedule for citrus fruit from Africa so that trade would not be interrupted while protecting U.S. agriculture from this invasive pest.     

Hot-water immersion quarantine treatment against Mediterranean fruit fly and Oriental fruit fly (Diptera: Tephritidae) eggs and larvae in litchi and longan fruit exported from Hawaii

Immersion of litchi fruit in 49 degrees C water for 20 min followed by hydrocooling in ambient (24 +/- 4 degrees C) temperature water for 20 min was tested as a quarantine treatment against potential infestations of Mediterranean fruit fly, Ceratitis capitata (Wiedemann); and oriental fruit fly, Bactrocera dorsalis Hendel, eggs or larvae in Hawaiian litchi, Litchi chinensis Sonnerat. The 49 degrees C hot-water immersion of litchi provided probit 9 (99.9968% mortality with >95% confidence) quarantine security against eggs and first instars. There were no survivors from 15,000 each feeding and nonfeeding Mediterranean fruit fly or oriental fruit fly third instars immersed in a computer-controlled water bath that simulated the litchi seed-surface temperature profile during the 49 degrees C hot-water immersion treatment. Litchi served as the model for longan, Dimocarpus longan Lour., a closely related fruit that is smaller and also has commercial potential for Hawaii. Modified fruit infestation and holding techniques used to obtain adequate estimated treated populations from poor host fruit, such as litchi and longan, are described. Data from these experiments were used to obtain approval of a hot-water immersion quarantine treatment against fruit flies for litchi and longan exported from Hawaii to the U.S. mainland.     

Effects of methyl bromide concentration, fumigation time, and fumigation temperature on Mediterranean and oriental fruit fly (Diptera: Tephritidae) egg and larval survival

The effects of methyl bromide (MB) concentration (16, 32,48, or 64 g/m3), fumigation temperature (15, 20, 25, or 30 degrees C), and fumigation time interactions on the survival of Mediterranean fruit fly, Ceratitis capitata (Wiedemann), and oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), eggs and first and third instars were recorded. Increasing the fumigation temperature from 15 to 20 degrees C or from 20 to 25 degrees C resulted in a significant reduction in fumigation time required for equivalent egg and larval mortalities at all studied MB concentrations; no further reductions in fumigation time resulted from increasing the temperature from 25 to 30 degrees C. Conversely, increasing temperature and time allowed for a reduction in MB concentration to obtain equivalent mortality. Thus, the optimum fumigation temperature for Mediterranean and oriental fruit fly eggs and larvae was 25 degrees C. Reducing MB concentrations required for phytosanitary fumigations would save time and expense, and reduce the amount of MB released into the atmosphere during aeration. Mediterranean fruit fly was as or more tolerant to MB than oriental fruit fly in MB tolerance for eggs and first instars. The egg stage was generally more tolerant to MB regardless of concentration. However, Mediterranean fruit fly eggs showed similar tolerance to first instars at 25 degrees C for the three highest concentrations and to third instars at 25 and 30 degrees C for the highest concentration, with no significant difference between them. Therefore, eggs alone can be used to obtain MB fumigation efficacy and quarantine security data at fumigation temperatures between 15 and 30 degrees C for Mediterranean and oriental fruit fly.     

Evaluation of the efficacy of the methyl bromide fumigation schedule against Mexican fruit fly (Diptera: Tephritidae) in citrus fruit

Methyl bromide fumigation is widely used as a phytosanitary treatment. Mexican fruit fly, Anastrepha ludens (Loew) (Diptera: Tephritidae), is a quarantine pest of several fruit, including citrus (Citrus spp.), exported from Texas, Mexico, and Central America. Recently, live larvae have been found with supposedly correctly fumigated citrus fruit. This research investigates the efficacy of the previously approved U.S. Department of Agriculture-Animal and Plant Health Inspection Service treatment schedule: 40 g/m3 methyl bromide at 21-29.4 degrees C for 2 h. Tolerance ofA. ludens to methyl bromide in descending order when fumigated in grapefruit (Citrus X paradisi Macfad.) is third instar > second instar > first instar > egg. Two infestation techniques were compared: insertion into fruit of third instars reared in diet and oviposition by adult A. ludens into fruit and development to the third instar. Inserted larvae were statistically more likely to survive fumigation than oviposited larvae. When fruit were held at ambient temperature, 0.23 +/- 0.12% of larvae were still observed to be moving 4 d postfumigation. Temperatures between 21.9 and 27.2 degrees C were positively related to efficacy measured as larvae moving 24 h after fumigation, pupariation, and adult emergence. Coating grapefruit with Pearl Lustr 2-3 h before fumigation did not significantly affect the proportion of third instars moving 24 h after fumigation, pupariating, or emerging as adults. In conclusion, fumigation with 40 g/m3 methyl bromide for 2 h at fruit temperatures >26.7 degrees C is not found to be inefficacious for A. ludens. Although a few larvae may be found moving >24 h postfumigation, they do not pupariate.     

Temperature thresholds and degree-day model for Marmara gulosa (Lepidoptera: Gracillariidae)

The developmental thresholds for Marmara gulosa Guillén & Davis (Lepidoptera: Gracillariidae) were investigated in the laboratory by using 17, 21, 25, 29, and 33 degrees C. The lowest mortality occurred in cohorts exposed to 25 and 29 degrees C. Other temperatures caused >10% mortality primarily in egg and first and second instar sap-feeding larvae. Linear regression analysis approximated the lower developmental threshold at 12.2 degrees C. High mortality and slow developmental rate at 33 degrees C indicate the upper developmental threshold is near this temperature. The degree-day (DD) model indicated that a generation requires an accumulation of 322 DD for development from egg to adult emergence. Average daily temperatures in the San Joaquin Valley could produce up to seven generations of M. gulosa per year. Field studies documented two, five, and three overlapping generations of M. gulosa in walnuts (Juglans regia L.; Juglandaceae), pummelos (Citrus maxima (Burm.) Merr.; Rutaceae), and oranges (Citrus sinensis (L.) Osbeck; Rutaceae), for a total of seven observed peelminer generations. Degree-day units between generations averaged 375 DD for larvae infesting walnut twigs; however, availability of green wood probably affected timing of infestations. Degree-day units between larval generations averaged 322 for pummelos and 309 for oranges, confirming the laboratory estimation. First infestation of citrus occurred in June in pummelo fruit and August in orange fruit when fruit neared 60 mm in diameter. Fruit size and degree-day units could be used as management tools to more precisely time insecticide treatments to target the egg stage and prevent rind damage to citrus. Degree-day units also could be used to more precisely time natural enemy releases to target larval instars that are preferred for oviposition.     

Quarantine strategies for olive fruit fly (Diptera: Tephritidae): low-temperature storage, brine, and host relations

A dose-response relationship was not observed in olive fruit fly, Bactrocera oleae (Gmelin), larvae exposed to acetic acid concentrations (0-2.5%) used in commercial brine solutions to cure olives. Immersion in a 1% acetic acid brine solution impeded emergence of the immature stages. A 1-wk exposure of olives infested with olive fruit fly larvae to low-temperature storage as a postharvest treatment at 0-1 degree C resulted in 8% survival of the population, and exposures of 2 through 5 wk further reduced pupal and adult emergence to <1.0%. One- to 2-wk exposures at 2-3 degrees C resulted in a significant decrease in survival from 20 to 3%, respectively, and longer durations of 3-5 wk reduced survival to <1.0%. Mean daily fruit pulp temperatures in olives in the top, middle, and bottom of plastic bins stored at 2-3 degrees C decreased by 5-8 degrees C from the first to the second day. Lowest temperatures were observed in the top, and highest temperatures were observed in the middle layer of fruit, which attained a mean temperature of 3.8 degrees C on day 5. Laboratory choice tests showed that olive fruit fly oviposited at a higher rate in late season Mission olives that were green than in fruit that were in the red blush maturity stage in tests with 1- and 3-4-d exposure periods, and an increase in duration of exposure was related to an increase in the total number of ovipositional sites. Higher percentages of olive fruit fly third instars, pupae, and adults were reared from green fruit than from fruit in the red blush stage after a 1-d exposure to oviposition. Manzanillo olives were more attractive for oviposition by olive fruit fly than Mission olives, and significantly more third instars, pupae, and adults developed in Manzanillo fruit than in Mission fruit in the red blush stage. These differences were related to the better quality and higher flesh content of the Manzanillo versus Mission olives used in the tests.     

Pre- and postharvest effects of lufenuron on Epiphyas postvittana (Lepidoptera: Tortricidae)

First-, third-, and fifth-instar Epiphyas postvittana (Walker) were exposed to a range of lufenuron concentrations (0-200 ppm) incorporated into synthetic diet and their subsequent development and mortality responses were determined. For all instars the greatest change in mortality response occurred over lufenuron concentrations < or = 3 ppm. However, third and fifth instars displayed an increase in mortality earlier than first instars, and were more sensitive to the lower lufenuron concentrations in this range. Only first and third instars subjected to < or = 2.5 ppm lufenuron survived the 26-d exposure trial. No larvae first exposed to lufenuron as first or third instars survived to pupation if ingesting concentrations of > or = 1 and > or = 3 ppm, respectively. Consumption of lower lufenuron concentrations by these larvae delayed pupation and resulted in pupal deformity. In contrast, fifth instars subjected to 100 ppm were capable of surviving the 26-d trial period and displayed a slower progressive reduction in survival to pupation with increase in lufenuron concentration. Also in contrast to more immature stages, fifth instars exposed to lufenuron developed more rapidly to pupation than larvae not exposed to the insect growth regulator (IGR), and all resulting pupae were normal. Third instars were exposed to sublethal lufenuron concentrations (0-3 ppm) for 4 d and the fourth-instar survivors subjected to a controlled atmosphere cold storage treatment (2% O2, 2% CO2, 0.6 degree C). Larvae ingesting diet containing 0.5 ppm (and to a lesser extent 1 ppm) lufenuron required longer exposure to the postharvest treatment to achieve > or = 95% mortality than larvae not ingesting the IGR. However, the analogous mortality response of larvae exposed to 3 ppm lufenuron was comparable to the control.     

Effect of cold treatment on survival and development of codling moth (Lepidoptera: Tortricidae) in cherry

'Bing' cherries, Prunus avium L., were obtained from an organic orchard and a conventional commercial orchard. The two groups were examined separately in replicated tests infested with each fruit initially infested with a first-instar codling moth, Cydia pomonella (L.). To simulate commercial postharvest holding conditions, the treatments were 0 (control), 1, 2, 4, 7, 10, and 14 d cold storage at 3.3 degrees C. The fruits were examined three or more times to determine larval survival, life stage, fruit condition, and fungal disease. Survival of first instars was affected only by cold storage durations of > or = 7 d. When infested with codling moth larvae, both organic and conventionally grown cherries quickly deteriorated from fungal diseases. The rate of moth development was estimated from the surviving larvae and was significantly different between organic and conventionally grown cherries for all instars except the second.     

Hot water immersion to ensure quarantine security for Cryptophlebia spp. (Lepidoptera: Tortricidae) in lychee and longan exported from Hawaii.

We determined whether immersion in 49 degrees C water for 20 min, a quarantine treatment developed for disinfestation of fruit flies in lychee, Litchi chinensis Sonn., and longan, Dimocarpus longan (Lourd.) Steud., exported from Hawaii, would also disinfest fruit of two species of Cryptophlebia. The pattern of tolerance to heat in Cryptophlebia illepida (Butler) was generally eggs < neonates < early instars = late instars < pupae. No C. illepida survived immersion for 16 or 20 min. Late fourth and fifth instars were determined to be the most tolerant stage that occurs in harvested fruit. Late instars of Cryptophlebia ombrodelta (Lower) were more tolerant of hot-water immersion than those of C. illepida, but no C. ombrodelta late instars survived immersion for 16 or 20 min. The hot water immersion quarantine treatment for fruit flies should effectively disinfest lychees and longans of any Cryptophlebia.     

Cold Storage of Coleomegilla maculata larvae

Short-term storage of second and third instars of the coccinellid Coleomegilla maculata lengi Timberlake can be achieved efficiently through utilization of temperatures below its thermal threshold of development. Survival, voracity and preimaginal development time were evaluated for larvae kept at 4 and 8°C for up to 5 weeks. Survival was close to 100% for the first two weeks of storage, but decreased drastically afterward and was 0% after 5 weeks. Storage at low temperatures did not diminish the voracity of larvae after storage. Development completely stopped during cold storage but resumed without significant effect after larvae were returned to 24°C. This study indicates for the first time that cold storage of C. maculata second and third instars is possible for a period of up to 2 weeks and provides a technique that could benefit biological control programs by increasing availability of beneficial insects.     

Psyttalia cf. concolor (Hymenoptera: Braconidae) for biological control of olive fruit fly (Diptera: Tephritidae) in California

The larval parasitoid, Psyttalia cf. concolor (Szépligeti), reared on Mediterranean fruit fly, Ceratitis capitata (Weidemann), by the USDA-APHIS-PPQ, Guatemala City, Guatemala, was imported into California for biological control of olive fruit fly, Bactrocera oleae (Gmelin), in olives, Olea europaea L. Mean percentage parasitism of olive fruit fly third instars infesting fruit in field cages ranged from 7.0 in Grapevine to 59.7 in Santa Barbara and in free releases ranged from 0 in Grapevine to 10.6 in Santa Barbara after 4- to 6-d exposures. In the laboratory, more parasitoids developed to adults in olive fruit fly larvae that were 11-13 d old than in larvae 8-10 d old. Adult parasitoids lived significantly longer when provided with water than adults without water in environmental chambers at 5 degrees C, 85% RH; 15 degrees C, 65% RH; 25 degrees C, 25% RH; and 35 degrees C, 25% RH. Adult parasitoids lived for 48 d with honey for food and water and 32 d with food and sugar solution at 15 degrees C and 65% RH. Survival of adult parasitoids without food and water in greenhouse tests was approximately 4 d in a simulated coastal climate and 1 d in a simulated inland valley climate and was significantly increased by providing food and water. The parasitoid did not develop in the beneficial seedhead fly, Chaetorellia succinea (Costa), in yellow star thistle. The rate of parasitism of walnut husk fly, Rhagoletis completa Cresson, larvae in green walnut husks was 28.4% in laboratory no-choice tests. In choice tests, the rate of parasitism of walnut husk fly versus olive fruit fly larvae in olives was 11.5 and 24.2%, respectively.     

Overwintering,soil distribution and phenology of Childers canegrub,Antitrogus parvulus (Coleoptera: Scarabaeidae) in Queensland sugarcane

In this study, the question of whether Childers canegrub, Antitrogus parvulus (Britton) overwinters in the subsoil was addressed. Irrigated fields of sugarcane were sampled during a 2-year period near Bundaberg in southern Queensland. Antitrogus parvulus overwintered as second and third instars at each of three sites. During autumn and winter third instars of different allochronic (separated in age by 12 months) populations occurred together and could not be readily separated. Field-collected third instars were reared on ryegrass and separated into two age groups based on the date of pupation. Third instars in the first year of their life cycle (young third instars) remained at shallow depth (100-200 mm) and did not overwinter in the subsoil as once thought. Minimum temperatures during winter were 13-16 degrees C and did not prevent young third instars from feeding and gaining weight. Third instars in their second and final year moved downwards from late summer and pupated in the subsoil at 293-425 mm in spring. General phenology was as previously reported with first instar larvae occurring from January until April, second instars from January until November and third instar larvae throughout the year. Prepupae and pupae were found between October and December and adults occurred in soil during November and January. Batches of eggs occurred at a mean depth of 350 mm. First and second instars occurred predominantly at relatively shallow (100-200 mm) depths in the soil profile. All stages tended to be most common under rows of sugarcane rather than in the interrow.     

Mechanisms contributing to the competitive success of the invasive fruit fly Bactrocera invadens over the indigenous mango fruit fly, Ceratitis cosyra: the role of temperature and resource pre-emption

We investigated the influence of temperature and infestation sequence on interspecific competition between two fruit flies: an invasive ( Bactrocera invadens Drew, Tsuruta & White, ( B ) and a native ( Ceratitis cosyra Walker , C ) (both Diptera: Tephritidae) species. Mango fruits [ Mangifera indica L. (Anacardiaceae)] were co-infested with larvae at different constant temperatures (15, 20, 25, and 30 °C) and relative humidity of 50 ± 8%, using different infestation sequences at each temperature ( BC together; BC/CB 1, 2, and 3 days apart). There were significant effects of competition in most experimental treatments, resulting in reduced larval survival, pupal mass, and adult emergence for both species. At most of the infestation/temperature combinations, C. cosyra was clearly the inferior competitor. The only exception was at 20 °C when the outcome depended on the sequence of infestation: no C . cosyra survived when the sequence was BC , but more C . cosyra than B . invadens survived when it was CB . At 15 °C, all C. cosyra larvae died, while the development of B. invadens was prolonged and adult emergence reduced. We conclude that resource pre-emption and fluctuations in temperature in mango agroecosystems help to explain observed shifts in dominance between B. invadens and C. cosyra on mango in many parts of Africa. The small window of competitive superiority for C. cosyra at 20 °C and CB infestation sequence, together with other factors such as fecundity and alternative hosts, may allow for co-existence in some environments.     

Thermal death kinetics of egg and third instar Mediterranean fruit fly (Diptera: Tephritidae)

Two developmental stages of Ceratitis capitata (Wiedemann), 24-h-old eggs and third instars, 8 d after oviposition, were subjected to thermal exposures in a heating block system, at various temperatures of 46, 48, 50, and 52 degrees C to determine the thermal death kinetics of the insects. At these temperatures, 100% mortality was achieved by exposure of 300 C. capitata larvae for 60, 15, 4, and 1 min, respectively. The 0.5 order kinetic model had the best fit to the survival ratio for all the treatment temperatures, hence it was used for the prediction of the lethal times. The thermal death time (TDT) curves showed that the third instars were more heat-resistant than eggs, especially at the two low temperatures (46 and 48 degrees C). Under temperature-time combinations that did not result in complete kill, the thermal mortality for eggs was also significantly higher than that for third instars. The activation energy values calculated from the TDT curves were 490.6 and 551.9 kJ/mol, respectively, for thermal death of eggs and third instars.     

Cold treatment for guava fruits infested with oriental fruit fly,Bactrocera dorsalis (Diptera: Tephritidae)

Guava (Psidium guajava Linnaeus) is an important horticulture crop in tropical and subtropical regions. However, guava transportation to the fruit market is regulated due to the risk of fruit fly infestations. The oriental fruit fly [Bactrocera dorsalis (Hendel), Tephritidae, Diptera], which damages numerous horticulture fruits, is the primary pest in guava. For fruit export, phytosanitary treatment is necessary to prevent the potential oriental fruit fly infestations through pest-infested fruits. In this study, cold treatment was administered to eliminate oriental fruit fly infestations in guava fruits. The optimal egg inoculation density was 200 eggs per fruit, and third instar larvae displayed the highest cold treatment tolerance. On assessing the efficacy of cold treatment, none of the third instar larvae in the fruits survived at a constant fruit core temperature of 0.5–1 °C for 11 days. A confirmatory test revealed that on maintaining a fruit core temperature of 0.5–1 °C for 12 days, 100% mortality was achieved among the fruit flies. In fruit quality assessment, guava fruits exposed to cold treatment still maintained their market value. Thus, cold treatment effectively disinfested guava fruits with possible fruit fly infestations, making it a viable quarantine treatment for guava fruits prior to their export.     

Sensitivity of wild and genetic sexing strain Ceratitis capitata to high-temperature disinfestation of Valencia oranges

Abstract:  The purpose of this research was to investigate post-harvest heat treatment of Valencia oranges as an effective disinfestation protocol (fast, no fruit damage) for the Mediterranean fruit fly Ceratitis capitata (Wiedemann). Forced high-temperature air was applied under the following conditions: (a) exposure to air temperature of 56°C, for fast increase of temperature in the interior of the fruit, (b) reduction in air temperature to 47°C when the fruit centre reached 47°C and (c) maintenance of fruits in the chamber for another 30 min. Relative humidity in the treatment chamber was kept between 50% and 65% during treatment. Forced air at 47°C applied for 30 min on eggs before hatch or late third instar larvae (the most heat-tolerant stages) resulted in complete kill. Egg and larval sensitivity to high temperature differed between a wild strain and a laboratory genetic-sexing strain based on white pupa mutation. In this strain males emerge from brown pupae and females from white pupae. In particular, mature eggs from the wild strain were significantly more temperature resistant than eggs from the laboratory strain. Exposure of Valencia oranges of a diameter of 7–7.5 cm to 56°C forced air for about 86 and 99 min was required to increase temperature to 47°C at 1.5 cm depth and the fruit centre respectively. Treated oranges showed no substantial peel or interior deterioration, or change in colour and taste when kept at 25°C and 50–60% RH for a period of up to 1 month following treatment. Treatment in 1% O₂ atmosphere, produced by flushing of CO₂ into the treatment chamber, resulted in about 1°C reduction in killing temperature and faster increase in temperature inside the fruit to a lethal level.     

Field infestation, life history and demographic parameters of the fruit fly Bactrocera invadens (Diptera: Tephritidae) in Africa

Field infestation rates of an invasive fruit fly species, Bactrocera invadens Drew Tsuruta & White on mango was determined at different localities in Kenya. At most of the locations and especially at low elevations, B. invadens frequently shared the same fruit with the indigenous fruit fly species Ceratitis cosyra (Walker) but often occurred at higher numbers than C. cosyra. The level of infestation varied with location ranging from 3.0 to 97.2 flies per kg of fruit. There was a significant inverse relationship between numbers of flies per kg of fruit and elevation at which fruit was collected, suggesting that B. invadens is a predominantly lowland pest. On an artificial diet, development of B. invadens immatures lasted 25 days; egg incubation required 1.2 days, larval development 11.1 days and puparia-adult development 12.4 days. About 55% of eggs developed to the adult stage. Life expectancy at pupal eclosion was 75.1 days in females and 86.4 days in males. Average net fecundity and net fertility were 794.6 and 608.1 eggs, respectively, while average daily oviposition was 18.2 eggs. Daily population increase was 11% and mean generation time was 31 days. Results are discussed in relation to the biology and ecology of the insect and in the development of mass rearing and control measures for B. invadens.     

Irradiation disinfestation of apple maggot (Diptera: Tephritidae) in hypoxic and low-temperature storage

Apple maggot, Rhagoletis pomonella (Walsh), is a quarantine pest of apples, Malus domestica Borkhausen, and pears, Pyrus communis L., shipped from much of the United States and Canada. As such, these fruits shipped from infested areas to uninfested areas must undergo a quarantine disinfestation treatment. The objective of this research was to develop irradiation quarantine treatments against apple maggot considering that fruit hosts may be stored under hypoxic or cold conditions when they are irradiated. Hypoxia increased from 30.5 to 35.7 Gy (17%) the estimated dose to achieve 99% prevention of the full pupal stage from irradiated third instars in apples compared with ambient atmospheres. However, 50 Gy completely prevented the full pupa in 22,360 and 15,530 third instars, respectively, irradiated in apples in ambient and hypoxic atmospheres. There was no difference in development to the full pupal stage in apple maggot third instars held at 1 or 24 degrees C when irradiated with 20 Gy. Because the maximum dose measured when 50 Gy was sought was 57 Gy, the latter should be the dose recommended for quarantine disinfestation of host fruits of the apple maggot. Apples and pears tolerate much higher doses.     

Tetraploid Induction by Inhibiting Mitosis I with Heat Shock, Cold Shock, and Nocodazole in the Hard Clam Mercenaria mercenaria (Linnaeus, 1758)

Tetraploid induction by inhibiting mitosis I with heat shock (32, 35, and 38°C), cold shock (1, 4, and 7°C), and nocodazole (0.02 to 1.6 mg/L) was investigated in the hard clam Mercenaria mercenaria. All treatments were applied to fertilized eggs about 5 min before the first cell division at 22 to 23°C, and lasted for 10, 15, and 20 min. Three replicates were produced for each treatment with different parents. The ploidy of resultant larvae and juveniles was determined with flow cytometry. Heat shock of 35 and 38°C was effective in inhibiting mitosis I, producing 54% to 89% tetraploid larvae. Heat shock of 32°C accelerated embryonic development without inhibiting mitosis or producing tetraploids. In all heat-shock groups, the survival to D-stage larvae was lower than in controls, suggesting that heat-shock treatments and tetraploidy were detrimental to larval development. At the juvenile stage, survivors from heat-shock groups contained no tetraploids. Cold shocks suspended the first cell division during the treatment, but produced no tetraploids in the 4°C and 7°C treatment groups. Cold shock of 1°C produced 31% tetraploid larvae in one replicate, with none surviving to juvenile stage. Nocodazole inhibited mitosis I at concentrations of 0.04 mg/L or higher, but did not produce tetraploids. This study indicates that heat shock is most effective in inducing tetraploids through mitosis I inhibition, although none of the induced tetraploids survived to juvenile stage.