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A systems approach was developed as an alternative to a standalone quarantine disinfestation treatment for Thaumatotibia leucotreta in citrus fruit exported from South Africa. The systems approach consists of three measures: pre and postharvest controls and measurements, postpacking inspection, and postharvest exposure to low temperatures. Different cold treatment conditions with a range of efficacy levels can be used for this last measure. A series of trials reported here evaluated the efficacy of seven temperatures ranging from 0 to 5°C for durations from 14 d to 26 d. Mortality of the most cold-tolerant larval stages of T. leucotreta was determined. Temperatures of 0, 1, 2, and 3°C for 16, 19, 20, and 24 d respectively, induced 100% mortality of the tested populations. Probit 9 level treatment efficacy was achieved at 0 and 1°C for 16 and 19 d respectively. Mortalities higher than 90% were obtained with temperatures of 4, 4.5, and 5°C, after exposure for the longer durations. We demonstrated a significant difference in cold-induced insecticidal efficacy between 1, 2, 3, and 4°C. There was no significant difference in insecticidal efficacy between 4 and 4.5°C, but both of these temperatures were more efficacious than 5°C. The results of this study are valuable to support the use of cold treatment conditions with lower risk of fruit chilling injury in an effective systems approach, where the cold treatment efficacy can be augmented with other components of the systems approach.

Bactrocera dorsalis (Hendel) is a new fruit fly pest of some fruit types in the north and north eastern areas of South Africa. In order to determine whether existing cold disinfestation treatment schedules for an indigenous fruit fly pest: Ceratitis capitata (Wiedemann) would be effective for B. dorsalis, cold tolerances of four immature stages of the two species were compared. Studies were done in an artificial carrot-based larval diet. The developmental rates of the immature stages of the two species in the carrot-based larval diet were first determined at a constant temperature of 26°C. The developmental times for eggs and three larval stages were found to be similar for the two species. Incubation times of both species after egg inoculation were determined to be 0, 3, 4, and 6 d for obtaining egg, first larval, second larval, and third larval stages respectively for the cold treatment. At a test temperature of –0.6°C, mortality rates of C. capitata eggs, first instars, second instars, and third instars were lower than those of B. dorsalis. These results demonstrate that the current cold treatment schedules for disinfestation of C. capitata can be used as equally or more efficacious treatments for B. dorsalis.

A systems approach was previously developed for mitigating phytosanitary risk of Thaumatotibia leucotreta (Meyrick) in citrus fruit exported from South Africa, as an alternative to a standalone cold disinfestation treatment. The present study first tested the original systems approach by applying it on a semicommercial scale in 10 Nova mandarin orchards. Fruit were inspected at points in the production, packing, and simulated shipping process, to assess performance of the systems approach. Additional data were obtained from 17 Valencia orange orchards and six packinghouses. In the second part of this study, the systems approach was accordingly revised and improved, consisting of three measures: 1) preharvest controls and measurements and postpicking sampling, inspection, and packinghouse procedures; 2) postpacking sampling and inspection; and 3) shipping conditions. The model quantifying the effectiveness of the systems approach was improved by correcting errors in the original version, updating parameter values and adding a component that provides for comparison with the risk mitigation provided by a standalone disinfestation treatment. Consequently, the maximum potential proportion of fruit that may be infested with live T. leucotreta after application of the improved systems approach is no greater than the proportion of fruit that may be infested after application of a Probit 9 efficacy postharvest disinfestation treatment to fruit with a 2% pretreatment infestation. The probability of a mating pair surviving is also determined. The model enables a priori determination of the required threshold levels for any of the three measures, based on quantification of the other two measures.

Some of South Africa's export markets require postharvest cold treatment of citrus fruit for phytosanitary risk mitigation for Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae). An alternative to a standalone cold treatment may be a reduced intensity cold treatment as a step in a systems approach. For cold treatment trials, large numbers of larvae are required. Due to recent dramatic improvement of T. leucotreta control in the field, sufficient naturally infested citrus fruit are no longer available. Artificial infestation of fruit is not viable due to rapid decay of the fruit. Consequently, it is necessary to use laboratory-reared T. leucotreta larvae in artificial diet. In trials, field-collected larvae from the Eastern Cape were at least as cold-tolerant as those from other regions. Larvae in Navel oranges showed the median level of susceptibility in a range of citrus types evaluated at 6°C, and their use in trials was considered acceptable due to their greater natural susceptibility to T. leucotreta infestation. We demonstrated that larvae at high density in artificial diet were at least as cold-tolerant as larvae at lower densities. When exposed to 2°C for 18 d or longer, larvae in artificial diet as used in the trials were at least as cold-tolerant as larvae in fruit. Very few surviving larvae from fruit completed development, with no subsequent generation. Consequently, it is considered justifiable to conduct cold-treatment trials with laboratory-reared T. leucotreta larvae in artificial diet without risk of overestimating the effect of cold on feral larvae in citrus fruit. [corrected]

Some of South Africa's citrus export markets require mandatory postharvest cold treatment of citrus fruit as a phytosanitary risk mitigation treatment for Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae). An alternative to this may be partial cold treatment as one of the final steps in a systems approach to mitigate phytosanitary risk. Consequently, the efficacy of such partial cold treatments was evaluated. It was first determined that a 2°C cold treatment was significantly more effective against fourth and fifth instars (the most cold-tolerant instars) than treatments at 3°C and 4°C for a duration of 18 d. Secondly, it was determined that 2°C for 18 d and 1°C for 16 d were similarly effective, but both treatments were significantly more effective than 1°C for 14 d. Mean mortality of fourth and fifth instars treated with 2°C for 18 d in seven replicates from four trials was 99.94%. Finally, it was determined that the inability of the majority of surviving larvae to develop to adulthood would further increase the efficacy of a 2°C for 18 d treatment to 99.96%. Inclusion of reproductive nonviability of survivors increased mortality to 99.99%.

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 temperatures0 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 temperatures0 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 temperatures1.4 degrees C, commencing once all fruit pulp probes reach a temperature of 1 degree C or lower.

A systems approach has been developed for mitigation of risk associated with Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), in citrus fruit exported from South Africa, as an alternative to a stand-alone cold treatment. This study was undertaken to assess compliance with inspection standards applicable to various steps within the systems approach and to determine its overall efficacy. Larval infestation of fruit was monitored weekly in fruit from 33 orchards, until the time of harvest, postpicking, and postpacking into export cartons. Significant positive regressions were recorded between infestation of fruit during the full monitoring period in the orchard and the last 4 wk before harvest, between the last 4 wk before harvest and on delivery to the packinghouse, and on delivery to the packinghouse and in the packed carton. There was an improvement in the level of compliance with each of these successive steps in the system, thus verifying that the grading and inspection thresholds were appropriately sensitive and confirmed the effectiveness of the system. The overall risk mitigation efficacy of the systems approach was calculated. The calculation included several known compounding under estimations of efficacy. Nonetheless, the proportion of fruit that could be infested with T. leucotreta after application of the systems approach was between P ≤ 5.328 × 10(-6) and P ≤ 8.380 × 10(-7), 6-38 times less than the proportion associated with the probit 9 (P ≤ 3.2 × 10(-5)) standard for a stand-alone cold treatment, being three survivors in 100,000 at the 95% confidence level.

To develop a cold disinfestation treatment for the fruit fly Bactrocera invadens Drew, Tsuruta & White (Diptera: Tephritidae) that is rapidly spreading across Africa, research was conducted in Nairobi, Kenya, using flies from a laboratory culture and 'Valencia' orange (Citrus sinensis L. Osbeck) as the host. The developmental rate of B. invadens in Valencia oranges was determined at 28 degrees C, and the third instar was found to be the least susceptible of the egg and larval life stages to cold treatment at 1.1 degrees C in oranges. When 22,449 B. invadens third instars were exposed in oranges to a cold treatment with an approximate midpoint of 1.1 +/- 0.5 degrees C, the results suggested that a period of 16 d would be worthwhile verifying on a larger scale in oranges. Results from the first replicate of 16,617 larvae showed no survivors, but the second replicate of 23,536 larvae had three survivors. Because a longer cold treatment based on a mean temperature of 1.1 degrees C would create logistical difficulties for some export markets, further replicates were conducted at an approximate midpoint of 0.5 degrees C and at mean hourly maximum of 0.9 +/- 0.5 degrees C, for 16 d. After three replicates, in which 65,752 B. invadens third instars in total were treated with no survivors, the Japanese requirement of 99.99% mortality at the 95% confidence level was surpassed. The following treatment protocol for B. invadens larvae in oranges can therefore be recommended: fruit pulp to be maintained at temperatures of 0.9 degrees C or lower for 16 consecutive days.