The papers presented in this issue are focused on developing and validating procedures to improve the overall quality of sterile fruit flies for use in area-wide integrated pest management (AW-IPM) programs with a sterile insect technique (SIT) component. The group was coordinated and partially funded by the Joint FAO/IAEA Programme of Nuclear Techniques in Food and Agriculture, International Atomic Energy Agency, Vienna, Austria, under a five-year Coordinated Research Project (CRP) on “Quality Assurance in Mass-Reared and Released Fruit Flies for Use in SIT Programmes”. Participants in the CRP from 16 countries came from both basic and applied fields of expertise to ensure that appropriate and relevant procedures were developed. A variety of studies was undertaken to develop protocols to assess strain compatibility and to improve colonization procedures and strain management. Specific studies addressed issues related to insect nutrition, irradiation protocols, field dispersal and survival, field cage behavior assessments, and enhancement of mating competitiveness. The main objective was to increase the efficiency of operational fruit fly programs using sterile insects and to reduce their cost. Many of the protocols developed or improved during the CRP will be incorporated into the international quality control manual for sterile tephritid fruit flies, standardizing key components of the production, sterilization, shipment, handling, and release of sterile insects.

The success of Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) control programs integrating the sterile insect technique (SIT) is based on the capacity of released the sterile males to compete in the field for mates. The Islands of Madeira are composed of 2 populated islands (Madeira and Porto Santo) where the medfly is present. To evaluate the compatibility and sexual performance of sterile flies we conducted a series of field cage tests. At same time, the process of laboratory domestication was evaluated. 3 wild populations, one semi-wild strain, and 1 mass reared strain were evaluated: the wild populations of (1) Madeira Island (north coast), (2) Madeira Island (south coast), and (3) Porto Santo Island; (4) the semi-wild population after 7 to 10 generations of domestication in the laboratory (respectively, for first and second experiment); and (5) the genetic sexing strain in use at Madeira medfly facility (VIENNA 7mix2000). Field cage experiments showed that populations of all origins are mostly compatible. There were no significant differences among wild populations in sexual competitiveness. Semi-wild and mass-reared males performed significantly poorer in both experiments than wild males in achieving matings with wild females. The study indicates that there is no significant isolation among strains tested, although mating performance is reduced in mass-reared and semi-wild flies after 7 to 10 generations in the laboratory.

This study documents differences in the courtship behavior of wild strains of Ceratitis capitata (Wiedemann) from Madeira (Portugal), Hawaii (U.S.A.), Costa Rica, and Patagonia (Argentina). Some traits showed large variations and others substantial overlaps. The angle at which the male faced toward the female at the moment of transition from continuous wing vibration and intermittent buzzing changed very little during the course of courtship in all strains, but males from Madeira tended to face more directly toward the female than other males. Females tended to look more, and more directly, toward the males as courtship progressed in all strains. The distance between male and female tended to decrease as courtship proceeded in all strains, but the distances at which males initiated continuous vibration, intermittent buzzing, and jumped onto the female were relatively less variable between strains, except for the strain from Costa Rica. Flies of Madeira courted for longer and the male moved his head and buzzed his wings longer than the other strains.

The mass-reared colony of Anastrepha ludens (Loew) currently used in Mexico for suppression of the Mexican fruit fly has been in use for over 10 years. Sterile flies are released into a wide range of environmental conditions as part of an integrated area-wide approach to suppress diverse populations of this pest in the Mexican Republic. This paper assesses the performance of the sterile flies interacting with wild populations from the different environments. We investigated the sexual compatibility and competitiveness of the sterile flies when competing with wild populations from 6 representatives Mexican states: Nuevo León, Tamaulipas, Sinaloa, Nayarit, Michoacán, and Chiapas. Results show that the males of the wild populations differed in the time to the onset and peak of sexual activity. Nevertheless, the index of sexual isolation (ISI) reflected sexual compatibility between the populations and the mass-reared strain, indicating that the sterile individuals mate satisfactorily with the wild populations from the 6 states. The male relative performance index (MRPI) showed that the sterile male is as effective in copulating as the wild males. The female relative performance index (FRPI) reflected a general tendency for wild females to copulate in greater proportion than the sterile females, except for the strains from Tamaulipas and Chiapas. In general, the lower participation of the sterile females in copulation increases the possibilities of sterile males to mate with wild females. The relative sterility index (RSI) showed that the acceptance by wild females of the sterile males (25-55%) was similar to that of wild males. Females of the Chiapas strain showed the lowest acceptance of sterile males. Finally, the results obtained in the Fried test (which measures induced sterility in eggs) showed a competitiveness coefficient ranging from 0.2 to 0.5. This suggests that sterile males successfully compete and are compatible with flies from different geographic origins.

We evaluated under semi-natural field cage conditions sexual compatibility and competitiveness of a laboratory strain (LAB) compared to a wild population (TUC) of Anastrepha fraterculus (Wiedemann). The LAB strain is produced under semi-mass rearing conditions at the Estación Experimental Agroindustrial Obispo Colombres facility (Tucumán, Argentina). Wild flies were obtained at Horco Molle (Tucumán, Argentina) from infested guava fruits. LAB pupae were irradiated (⁶⁰Co) 48 h before adult emergence. The tested doses were 0 (control), 40, 70, and 100 Gy. Twenty-five males and 25 females each of TUC and LAB were released into cages and mating pairs collected. Only 1 irradiation dose was considered at a time. Females were separated and allowed to lay eggs into artificial fruits to estimate induced sterility from the corresponding hatching rate. Copulation start time did not differ significantly between strains nor among irradiation treatments. Copulation duration showed highly significant differences among irradiation doses, but no differences between strains. The index of sexual isolation (ISI) and the relative sterility index (RSI) indices indicated that LAB and TUC are fully compatible, males from TUC and LAB did not differ in mating competitiveness, and irradiation within the range tested did not affect these indices. Non-irradiated LAB females exhibited higher mating propensity than TUC ones. However, a significant reduction in the female relative performance index (FRPI) index was observed with increasing irradiation dose. The analysis of induced sterility indicated that treatment with 40 Gy reduces male fertility from about 80% to 0.75%, and higher doses produce total sterility. In females, the 40 Gy dose reduces fertility to about 2% and higher doses prevent egg laying.

Mass rearing conditions affect the mating behavior of Mediterranean fruit flies (medflies) Ceratitis capitata (Wiedemann). We evaluated the effect of slight changes in the adult holding conditions of adult flies maintained for egg production on their mating performance. Colonization was initiated from wild flies collected as larvae from infested coffee berries (Coffea arabica L.). When pupae were close to adult emergence, they were randomly divided into 3 groups and the emerging adults were reared under the following conditions: (1) Metapa System (MS, control), consisting of 70 × 45 × 15 cm aluminum frame, mesh covered cages, with a density of 2,200 flies per cage and a 1:1 initial sex ratio; (2) Insert System (IS), with the same type of cage, and the same fly density and sex ratio as in the MS treatment, but containing twelve Plexiglas® pieces (23 × 8.5 cm) to provide additional horizontal surface areas inside the cage; and (3) Sex-ratio System (SS), same as IS, but in this case the initial male: female ratio was 4:1. Three d later, newly emerged females were introduced, so the ratio became 3:1 and on the 6th d another group of newly emerged females was added to provide a 2:1 final sex ratio, at which the final density reached 1,675 flies per cage. The eggs collected from each of the 3 treatments were reared independently following standard procedures and the adults were held under the same experimental conditions. This process was repeated for over 10 to 13 generations (1 year). The experiment was repeated 3 times in 3 consecutive years, starting each replicate with a new collection of wild flies. Life tables were constructed for each treatment at the parental, 3rd, 6th, and 9th generations. Standard quality control parameters (pupation at 24 h, pupal weight, adult emergence, and flight ability), were estimated for each treatment every third generation in the third year. For the last generation each year, mating competitiveness was evaluated in field cage tests with wild flies. As colonization progressed, life expectancy and fecundity rates increased in the 3 rearing systems. There was no significant difference in standard quality control parameters among the 3 rearing systems. Wild males always achieved more matings than any of the mass reared males. Mating competitiveness of males from the IS, although surprisingly not from the SS, was significantly greater than that of males from the MS. Our results indicate that these slight changes in the adult holding conditions can significantly reduce the harmful effects of mass rearing on the mating performance of sterile flies.

A mass-rearing facility for Mediterranean fruit fly Ceratitis capitata (Wiedemann) was commissioned in Stellenbosch in 1999 to produce sterile male fruit flies for a sterile insect technique (SIT) project in commercial fruit orchards and vineyards in the Western Cape province of South Africa. The mass-rearing procedure was largely based on systems developed by the FAO/IAEA Agriculture and Biotechnology Laboratory, Seibersdorf, Austria. A number of genetic sexing strains were used to produce only males for release. Initial cramped rearing and quality management conditions were alleviated in 2001 with the construction of a new adult rearing room and quality control laboratory. In 2002 a comprehensive Quality Management System was implemented, and in 2003 an improved genetic sexing strain, VIENNA 8, was supplied by the FAO/IAEA Laboratory in Seibersdorf. For most of the first 3 years the facility was unable to supply the required number of sterile male Mediterranean fruit flies for the SIT program without importing sterile male pupae from another facility. From mid-2002, after the quality management system was implemented, both production and quality improved but remained below optimum. After the introduction of the VIENNA 8 genetic sexing strain, and together with an improvement in the climate control equipment, production stability, and quality assurance parameters improved substantially. The critical factors influencing production and quality were an inadequate rearing infrastructure, problems with the quality of the larval diet, and the initial absence of a quality management system. The results highlight the importance of effective quality management, the value of a stable and productive genetic sexing strain, and the necessity for a sound funding base for the mass-rearing facility.

The integration of the sterile insect technique (SIT) in the management of the South American fruit fly Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) is a promising alternative to chemically-based control in those areas where it is sympatric with Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) or other tephritid species for which the SIT is being used. Implementation of the SIT requires the development of a cost effective mass-rearing protocol. In this work, we present demographic and quality control parameters for the A. fraterculus strain reared at the Estación Experimental Agroindustrial Obispo Colombres, Tucumán, Argentina. Considering the rearing cage as the reproduction unit, we observed that fecundity is optimal during the first 3 weeks after the onset of oviposition. Fertility was constant during this period. During 2003 and 2004, some improvements were made to the existing rearing protocol, which resulted in increased larval viability, pupal weight, and adult emergence. Current weekly egg production is 1 million per week. These eggs are used to maintain the colony and to assess quality parameters. Finally, research needs leading to improved yields and fly quality are discussed.

Quality control procedures for Bactrocera philippinensis Drew & Hancock 1994 (Diptera: Tephritidae) used in sterile insect technique (SIT) programs were established in the mass rearing facility at the Philippine Nuclear Research Institute. Basic studies on pupal irradiation, holding/packaging systems, shipping procedures, longevity, sterility studies, and pupal eye color determination in relation to physiological development at different temperature regimes were investigated. These studies will provide baseline data for the development of quality control protocols for an expansion of B. philippinensis field programs with an SIT component in the future.

Sterile male insects, mass-reared and released as part of sterile insect technique (SIT) programs, must survive long enough in the field to mature sexually and compete effectively with wild males for wild females. An often reported problem in Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) SIT programs is that numbers of released sterile males decrease rapidly in the field for various reasons, including losses to different types of predators. This is a serious issue in view that most operational programs release sterile flies at an age when they are still immature. Previous field and field-cage tests have confirmed that flies of laboratory strains are less able to evade predators than wild flies. Such tests involve, however, considerable manipulation and observation of predators and are therefore not suitable for routine measurements of predator evasion. Here we describe a simple quality control method with aspirators to measure agility in medflies and show that this parameter is related to the capacity of flies to evade predators. Although further standardization of the test is necessary to allow more accurate inter-strain comparisons, results confirm the relevance of measuring predator evasion in mass-reared medfly strains. Besides being a measure of this sterile male quality parameter, the described method could be used for the systematic selection of strains with a higher capacity for predator evasion.

The pattern of gas-exchange (CO₂ emission) was investigated for developing Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) pupae incubated at different temperatures. This study was undertaken to explore the usefulness of gas-exchange systems in the determination of physiological age in developing pupae that are mass produced for sterile insect technique projects. The rate of CO₂ emission was measured in a closed flow-through system connected to commercial infrared gas analysis equipment. Metabolic activity (rate of CO₂ emission) was related to pupal eye-color, which is the current technique used to determine physiological age. Eye-color was characterized digitally with 3 variables (Hue, Saturation and Intensity), and color separated by discriminant analysis. The rate of CO₂ emission throughout pupal development followed a U-shape, with high levels of emission during pupariation, pupal transformation and final pharate adult stages. Temperature affected the development time of pupae, but not the basic CO₂ emission patterns during development. In all temperatures, rates of CO₂ emission 1 and 2 d before adult emergence were very similar. After mid larval-adult transition (e.g., phanerocephalic pupa), digital eye-color was significantly correlated with CO₂ emission. Results support the suggestion that gas-exchange should be explored further as a system to determine pupal physiological age in mass production of fruit flies.

Irradiation of pupae in sterile insect technique (SIT) projects is usually undertaken in hypoxic atmospheres, which have been shown to lessen the deleterious effects of irradiation on the quality of adult sterile flies. Although this is the accepted technology in most mass-rearing and sterilization facilities, to date no information has been generated on the actual levels of oxygen (O₂) in pupae-packing containers during irradiation. The present study utilized recently-developed technology to investigate the O₂ level inside bags in which pupae of Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) are packed prior to irradiation, the ability of pupae to create hypoxic environments in these bags, and the effect of O₂ atmospheres on the quality of irradiated males. Pupae, 1 d before adult emergence, were shown to deplete the O₂ level in sealed bags in approximately 1 h. The rate of O₂ consumption was dependent upon pupal age and incubation temperature. Incubation temperature did not significantly affect the quality of pupae or mating capacity of resultant adult males if pupae were irradiated under maximal hypoxic conditions inside packing bags. In contrast, mating competitiveness drastically decreased when pupae were irradiated under ambient O₂ conditions, with the packing bag open. There was no difference in the mating capacity of males when pupae were irradiated in sealed bags under either 10% or 2% O₂ levels, or under maximal hypoxia. Normal doses of fluorescent dye, applied to pupae to mark sterile flies, did not affect the ability of pupae to create hypoxic conditions inside packing bags, nor the quality control parameters of either pupae or adults. Current practices in mass-rearing facilities are discussed in the light of these results.

The sterile insect technique (SIT) is an environment-friendly pest control technique with application in the area-wide integrated control of key pests, including the suppression or elimination of introduced populations and the exclusion of new introductions. Reproductive sterility is normally induced by ionizing radiation, a convenient and consistent method that maintains a reasonable degree of competitiveness in the released insects. The cost and effectiveness of a control program integrating the SIT depend on the balance between sterility and competitiveness, but it appears that current operational programs with an SIT component are not achieving an appropriate balance. In this paper we discuss optimization of the sterilization process and present a simple model and procedure for determining the optimum dose.

In relation to the application of the sterile insect technique (SIT) for the South American fruit fly Anastrepha fraterculus (Wiedemann), we analyzed the effect on adult fertility of different doses of gamma irradiation and the age of pupae at the time of irradiation. In a first experiment, we applied doses of 50, 70, and 90 Gy to pupae at 24, 48, 72, and 96 h before adult emergence. In a second experiment we irradiated pupae 48 h before emergence with 20, 40, and 60 Gy and estimated male and female fertility and sperm transfer by irradiated males. The results indicated pupal age at irradiation does not significantly affect male fertility. If males irradiated with 60 Gy are crossed to non-irradiated females the fertility is about 1%. Females irradiated with 40 Gy did not lay eggs independently of the male to which they mated. No significant effects of radiation were observed with respect to the ability of males to transfer sperm. A dose of 70 Gy applied 48 h before adult emergence induces 100% sterility in both males and females.

The operational use of Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann), genetic sexing strains in Sterile Insect Technique applications can be maximized by developing methods for effective shipment of eggs. This would enable a central production facility to maintain the relevant mother stocks and large colonies to supply eggs to satellite centers that would mass produce only males for irradiation and release. In order to achieve this, the survival of medfly embryos of different ages was assessed after storage at 5, 10, 15, 20, and 25°C in water for different periods of time. Survival was affected by all 3 variables, i.e., embryo age, water temperature, and length of storage. Storage of embryos at any temperature for 120 h resulted in almost no survival. Controlling the age of the embryo at the time of the temperature treatment is crucial for the success of this procedure. Embryos collected between 0 to 12 h after oviposition and pre-incubated at 25°C for 12 h provide a suitable 72 h window for shipment when maintained between 10 to 15°C. Under these conditions, no significant reductions in survival during all the developmental stages were observed.

The present study has been conducted to assess the effect and interaction of various storage substrates and conditions on eggs of the Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann). Tests were carried out with the genetic sexing strain VIENNA 8/D53, a strain that carries a temperature sensitive lethal (tsl) mutation that allows the selective killing of female zygotes. This study identifies strategies to enhance the storage and transport conditions through assessment of effect on egg, pupal and adult survival in order to facilitate the establishment of satellite mass rearing facilities for the production of male medflies. Eggs were immersed in two different substrates and stored at different temperatures and for different time periods. Findings from this study suggest that egg storage periods, and to some extent, the storage substrates have significant effects on pupal and adult survival. For 72-h storage periods, the eggs preserved in agar solution at 10°C produced the most pupae. There was an inverse relationship between the concentration of dissolved oxygen in the substrate during storage and the quality and survival of the stored/transported eggs. Apparently low levels of dissolved oxygen reduce metabolic rates, allowing the storage period to be prolonged.

The sterile insect technique (SIT) is increasingly being used around the world to control Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), the Mediterranean fruit fly as part of an area-wide integrated approach. One option that may improve the effectiveness of the SIT, by increasing the sexual competitiveness of released sterile males, consists of feeding males protein during the post-teneral stage, a diet that increases sexual performance of wild males. We examine the effects of diet on the successive hurdles males must overcome in order to inseminate females, i.e., joining leks, copulating females, having their sperm stored and inhibition of female remating. In addition, we address the effects of diet on post-release foraging success, longevity, and the ability to withstand starvation. While protein feeding universally increases the sexual success of wild males, its effect on sterile males varies with strain, experimental settings, and environmental conditions. In some cases, treatments that resulted in the best sexual performance were significantly associated with increased vulnerability to starvation. However, no particular diet affected the ability of sterile males to find nutrients in the field when these where available. We suggest it may be better to release relatively short-lived flies that are highly competitive, rather than long-lived, sexually ineffective ones.

The rates of dispersal and survival of sterile mass-reared laboratory flies and sterile wild flies of Anastrepha ludens (Loew) and Anastrepha obliqua (Macquart) were estimated and compared with a regular rectangular array of 64 food-baited traps spaced 60 m between traps around the release point in Tapachula Chiapas, Mexico. The traps were scored every day during the first week, and then every 3 d over a 30-d period. For A. obliqua, the number of males recaptured was higher than that of females, while for A. ludens, females were recaptured more frequently than males. The recapture rate for the wild strains ranged from 0.6-24.8% for A. ludens and 1.3-16.2% for A. obliqua and the corresponding ranges for the mass-reared strains were 0.5-7.1% and 0.5-3.0% respectively. The life expectancy was 4.7 d for wild and 4.3 d for mass-reared A. obliqua males but 3 and 2 d, respectively, for wild and mass-reared A. ludens males. The net displacement of A. ludens and A. obliqua ranged approximately from 100-250 m and took place mostly on the first day. Wild A. ludens moved to the northwest from the release point while the mass-reared strain moved to the west. The A. obliqua wild flies moved to the west, while the mass-reared strain shifted to the southwest. We discuss the implications of our findings as to the spacing and frequency of sterile fly releases for the suppression of wild populations.

Both wild and released (sterile) Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) and wild Bactrocera papayae (Drew and Hancock) in Australia had patchy distributions and comparisons with predictions of the negative binomial model indicated that the degree of clumping was sometimes very high, particularly at low densities during eradication. An increase of mean recapture rate of sterile B. tryoni on either of 2 trap arrays was not accompanied by a reduction in its coefficient of variation and when recapture rates were high, the percentage of traps catching zero decreased only slightly with increase in recapture rate, indicating that it is not practicable to decrease the heterogeneity of dispersion of sterile flies by increasing the number released. There was often a mismatch between the dispersion patterns of the wild and sterile flies, and the implications of this for the efficiency of the sterile insect technique (SIT) were investigated with a simulation study with the observed degrees of mismatch obtained from the monitoring data and assuming the overall ratio of sterile to wild flies to be 100:1. The simulation indicated that mismatches could result in the imposed rate of increase of wild flies being up to 3.5 times higher than that intended (i.e., 0.35 instead of 0.1). The effect of a mismatch always reduces the efficiency of SIT. The reason for this asymmetry is discussed and a comparison made with host-parasitoid and other systems. A release strategy to counter this effect is suggested.

The South American fruit fly Anastrepha fraterculus (Wiedemann) is one of the most destructive fruit pests in this region, infesting major fruit crops. Implementation of the sterile insect technique (SIT) as part of an area-wide integrated approach against this species requires information on the survival of mass-reared and sterilized insects in the field and their ability to mate with wild females. The survival rates in field cages of both non-irradiated and irradiated laboratory flies were compared with that of wild flies. Both types of laboratory flies survived longer than their wild counterparts over the 8 days under the experimental conditions. The irradiation dose (70 Gy) did not affect survival of the laboratory reared flies. Our results improve the prospect of integrating the SIT into the control of A. fraterculus populations in Argentina.

Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) displays a lek mating system. Males form groups in which they simultaneously display signals (acoustical, visual, or chemical) to attract females with the purpose of mating. Females visit the lek and choose among signaling and courting males to mate. Scarce information is available in A. fraterculus about the main factors involved in female choice and the behavior of displaying males. This information could be important within the context of pest control programs with a sterile insect technique (SIT) component, because departures from normal sexual behavior caused by artificial rearing could affect males’ performance in the field. In this study we assessed A. fraterculus male behavior within the leks and analyzed the importance of behavioral and morphological traits on their copulatory success. The existence of preferred places for lek formation was evaluated in field cages with trees inside and analyzed by dividing the trees in sectors according to a 3-dimensional system. Males were individually weighed, marked, and observed every 15 min. Morphometric and behavioral characteristics of successful and unsuccessful males were compared. Most successful males grouped in a region of the tree characterized by the highest light intensity in the first 2 h of the morning. Results showed that pheromone calling activity is positively associated with copulatory success. Copulations were more frequent for males calling inside the lek, indicating that pheromone calling activity and presence in the lek are key factors for copulatory success. A positive association between copulatory success and eye length was found; some characteristics of the face were also associated with copula duration and latency.

Despite the interest in applying environmentally friendly control methods such as sterile insect technique (SIT) against Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae), information about its biology, taxonomy, and behavior is still insufficient. To increase this information, the present study aims to evaluate the performance of wild flies under field cage conditions through the study of sexual competitiveness among males (sexual selection). A wild population from Horco Molle, Tucumán, Argentina was sampled. Mature virgin males and females were released into outdoor field cages to compete for mating. Morphometric analyses were applied to determine the relationship between the multivariate phenotype and copulatory success. Successful and unsuccessful males were measured for 8 traits: head width (HW), face width (FW), eye length (EL), thorax length (THL), wing length (WL), wing width (WW), femur length (FL), and tibia length (TIL). Combinations of different multivariate statistical methods and graphical analyses were used to evaluate sexual selection on male phenotype. The results indicated that wing width and thorax length would be the most probable targets of sexual selection. They describe a nonlinear association between expected fitness and each of these 2 traits. This nonlinear relation suggests that observed selection could maintain the diversity related to body size.

Application of the sterile insect technique (SIT) as part of integrated area-wide programs to control the Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) require that the released males attract wild females and transfer sterile sperm. However, knowledge about male sexual performance after they are released is scarce. We conducted a study to evaluate male sexual performance in field cage tests, according to standard quality control procedures. Mass-reared 5-d-old sterile males from the genetic sexing strain VIENNA 7mix2000 were acclimated for 0, 1, and 3 d to outdoor conditions before competing with wild males for wild females. Although the proportion of mating (PM) in the test was satisfactory, the resulting relative sterility index (RSI) data showed no significant differences among the treatments. The data indicate that pre-conditioning males to outdoor conditions in Madeira did not confer an advantage in field cage sexual performance.

The results of previous studies that showed that exposing mass-reared male Mediterranean fruit flies Ceratitis capitata (Wiedemann) to ginger root oil (“aromatherapy”) increases the likelihood of mating with wild females were confirmed. The increased male success could be due to female responses to changes in male behavior or male pheromones. There were no significant differences in the types of courtship movements executed by males with and without aromatherapy. The durations of movements also did not differ when mass-reared males were paired with mass-reared females; however, when they were paired with wild females, there were a few, small differences. Previous studies indicated that the effectiveness of the male long-distance attractant pheromone is not affected by aromatherapy, but these studies did not consider pheromones released at close range during courtship, which behavioral analyses suggest may be different. We propose the following possible explanation for the different effects of aromatherapy with different females. Selection on males under mass rearing may have altered their close-range pheromones in ways that can be remedied by aromatherapy; and only wild females respond because the pheromonal responsiveness of mass-reared females has also changed. We propose observations that could test these ideas.

The preimaginal stages of T. peregrina are described. The egg displays a sculptured chorion, which is found only on those deposited externally. The immature larvae are characterized by their peculiarities in (a) a setose ring on the thoracic and abdominal segments, (b) an anal notch and (c) size and the sensory structures of the head capsule. The final instar is described and illustrated. Morphological structures of diagnostic value are discussed. The most salient character shown by the mature larva of this species lies in the epistoma, which is complete.

Recently re-named as a sub-species of Mellinus arvensis, Mellinus arvensis obscurus Handlirsch 1888 was investigated ecologically and morphologically in Nepal, in order to underline the most important differences with the well known M. arvensis arvensis. Mellinus arvensis obscurus females nested in clumped aggregations on inclined plains at high altitudes, both on sunny bare soil and on a shaded grassy one. Beginning of monsoon season probably interfered with wasp activity, and females performed few provisioning flights during the day. Prey consisted of a broad range of Diptera, except for one case of a spider. Many females were observed not provisioning a nest but floating on the nesting site, and many intraspecific interactions suggested a high degree of usurpation attempts. At least one species of flies and two of ants probably acted as natural enemies of the wasp. Morphological observations on females showed that the Nepal population shares more similarities (shape of tergite I, body punctation) with the European populations than with the closer Japanese population; melanization is strong, according to west-east and altitudinal cline. The mature larva of M. arvensis obscurus Handlirsch is described, illustrated, and compared with the other mature larva of the genus. The differences between both larvae mainly lie in the presence/absence, and number or differentiation of integumental structures. We conclude that morphological traits are more important than ecological and behavioral ones in distinguishing M. arvensis obscurus from M. arvensis arvensis.

Host range of Xylosandrus mutilatus (Blandford) in North America is reported here for the first time. Descriptive data such as number of attacks per host, size of stems at point of attacks, and height of attacks above ground are presented. Hosts observed in Mississippi were Acer rubrum L., Acer saccharum Marsh., Acer palmatum Thunb., Ostrya virginiana (Mill.) K. Koch., Cornus florida L., Fagus grandifolia Ehrh., Liquidamber styraciflua L., Carya spp., Liriodendron tulipifera L., Melia azedarach L., Pinus taeda L., Prunus serotina Ehrh., Prunus americana Marsh., Ulmus alata Michaux, and Vitus rotundifolia Michaux. Liquidamber styraciflua had significantly more successful attacks, significantly higher probability of attacks, and significantly higher number of adult beetles per host tree than did Carya spp., A. rubrum, and L. tulipifera. This information is relevant in determining the impact this exotic beetle may have in nurseries, urban areas, and other forestry systems where this beetle becomes established.

The salivary glands of the Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis Germar 1821, (syn. H. coagulata, Hemiptera: Cicadellidae) were collected and used to produce a cDNA library. Examination by BLASTX analyses identified 2 viral sequences, one a 610-base pair fragment and a second 839-base pair fragment, both of which had significant homology to viruses within the genus Phytoreovirus. Resequencing of the fragments confirmed sequence validities. These sequences were used for in silico protein translation and BLASTP analysis confirming the established homology. While the GWSS is the primary vector of Pierce’s disease of grapes, this is the first report that GWSS may be a vector of a phytoreoviruses. Phylogenetic and homology comparisons with BLASTX, BLASTP, and PAUP analyses indicated that the viral sequences isolated from GWSS were closely related to the viruses in the Family Reoviridae, Genus Phytoreovirus, specifically Rice Dwarf Phytoreovirus (RDV). RDV is the only plant reovirus that is not limited to the phloem. Phytoreoviruses are transmitted in a propagative manner by cicadellid leafhoppers (Hemiptera: Cicadellidae), which acquire and transmit them during feeding. Phytoreoviruses have been reported from Agallian, Agalliopsis, Nephotettix, and Recilia, genera of leafhoppers, with evidence for transovarial transmission. The GWSS, although considered to feed primarily from the xylem, ingests from other plant tissues, such as the phloem and mesophyll during probing similar to other leafhoppers. The feeding behavior and wide host range of the GWSS provides an overlapping condition for these two organisms, leafhopper and virus. GWSS will feed from grasses as a transitory host, and on herbaceous and woody plants as primary hosts, which may favor the acquisition and transmission of Phytoreovirus by this leafhopper. Monitoring for an increase of Phytoreovirus spread in graminaceous crops that are in proximity to vineyards or tree crop orchards, where GWSS occurs, such as in southern California, will provide a better understanding of the potential role of the GWSS as a disease vector in the spread of phytoreoviruses and other plant pathogens. The sequences have been deposited in NCBI database under the accession numbers (EF058280) for GWSS-V1, WHSg013C11 and (EF058281) for GWSS-V2, WHSg024H02.

The parasitoid Lipolexis oregmae Gahan (introduced as L. scutellaris Mackauer) was imported from Guam, evaluated in quarantine, mass reared, and released into citrus groves in Florida in a classical biological control program directed against the brown citrus aphid, Toxoptera citricida Kirkaldy. Releases of 20,200, 12,100, and 1,260 adults of L. oregmae were made throughout Florida during 2000, 2001, and 2002, respectively. To determine if L. oregmae had successfully established, surveys were conducted throughout the state beginning in the summer of 2001 and continuing through the summer of 2003. Parasitism during 2001 and 2002 was evaluated by holding brown citrus aphids in the laboratory until parasitoid adults emerged. Lipolexis oregmae was found in 10 sites in 7 counties and 4 sites in 3 counties with parasitism rates ranging from 0.7 to 3.3% in 2001 and 2002, respectively. Laboratory tests indicated that high rates of mortality occurred if field-collected parasitized aphids were held in plastic bags, so a molecular assay was used that allowed immature L. oregmae to be detected within aphid hosts immediately after collection. The molecular assay was used in 2003 with the brown citrus aphids and with other aphid species collected from citrus, weeds, and vegetables near former release sites; immatures of L. oregmae were detected in black citrus aphids, cowpea aphids, spirea aphids, and melon aphids, as well as in the brown citrus aphid, in 4 of 8 counties sampled, with parasitism ranging from 2.0 to 12.9%, indicating that L. oregmae is established and widely distributed. Samples taken in Polk County during Oct 2005 indicated that L. oregmae has persisted. The ability of L. oregmae to parasitize other aphid species on citrus, and aphids on other host plants, enhances the ability of L. oregmae to persist when brown citrus aphid populations are low.

Lipolexis oregmae Gahan was introduced into Florida in a classical biological control program directed against the brown citrus aphid, Toxoptera citricida (Kirkaldy), on citrus. Prior to evaluating distribution, host range, and potential nontarget effects of L. oregmae in Florida, we evaluated the role of other potential host aphids and host plants on mummy production and location. Under laboratory conditions, this parasitoid produced the most progeny on the target pest, the brown citrus aphid on citrus. This parasitoid, unlike the majority of aphidiids, did not produce mummies on any of the host plants tested when reared in black citrus aphid T. aurantii (Boyer de Fonscolombe) on grapefruit, spirea aphid Aphis spiraecola Patch on grapefruit and pittosporum, cowpea aphid A. craccivora Koch on grapefruit and cowpeas, or melon aphid A. gossypii Glover on grapefruit and cucumber. Thus, sampling for L. oregmae mummies of these host aphids and host plants must involve holding foliage in the laboratory until mummies are produced. This parasitoid requires high relative humidity to produce adults because no adults emerged when mummies were held in gelatin capsules, but high rates of emergence were observed when mummies were held on 1.5% agar plates. In addition, we compared the color of 6 aphid hosts and the color of mummies produced by L. oregmae when reared in them to determine if color of mummies could be used to identify L. oregmae. Mummy color varied between aphid hosts and tested host plants, and is not a useful tool for identifying L. oregmae for nontarget effects.

An open-field experiment was conducted to assess the suitability of the South American leaf feeding beetle Gratiana boliviana Spaeth for biological control of Solanum viarum Dunal in the USA. An open-field test with eggplant, Solanum melongena L., was conducted on the campus of the University of Buenos Aires, Argentina, and a S. viarum control plot was established 40 km from the campus. One hundred adult beetles were released in each plot at the beginning of the experiment during the vegetative stage of the plants, and forty additional beetles were released in the S. melongena plot at the flowering stage. All the plants in each plot were checked twice a week and the number of adults, immatures, and eggs recorded. Results showed almost a complete rejection of eggplant by G. boliviana. No noticeable feeding damage was ever recorded on eggplant. The experiment was ended when the eggplants started to senesce or were severely damaged by whiteflies and spider mites. The results of this open-field experiment corroborate previous quarantine/laboratory host-specificity tests indicating that a host range expansion of G. boliviana to include eggplant is highly unlikely. Gratiana boliviana was approved for field release in May 2003 in the USA. To date, no non-target effects have been observed either on eggplant or native species of Solanum.

Non-indigenous invasive mole crickets, Scapteriscus vicinus Scudder (Orthoptera: Gryllotalpidae) in Florida and S. didactylus (Latreille) (the “changa”) in Puerto Rico, are being managed with an entomopathogenic nematode, Steinernema scapterisci (Nguyen and Smart) (Rhabditida: Steinernematidae), and a parasitic wasp, Larra bicolor L. (Hymenoptera: Sphecidae). Pest mole cricket populations have declined by 95% in north central Florida since these specialist natural enemies were released and established in the 1980s. Commercial production of the nematode was initiated, nearly 70 billion were applied in 34 Florida counties, and their establishment, spread, and impact on mole crickets were monitored. The infected mole crickets dispersed the nematode rapidly, so that within 6 months these parasites were present in most of the insects trapped in experimental pastures. Three years later, mole cricket populations were reduced to acceptable levels and the bahiagrass had recovered. The nematode was released for the first time in Puerto Rico during 2001 and has persisted; the wasp was introduced in the late 1930s. The geographical distribution of the wasp is being expanded in Florida and Puerto Rico by planting plots of Spermacoce verticillata (L.), a wildflower indigenous to Puerto Rico and widely distributed in southern Florida. Pastures, sod farms, golf courses, landscapes, and vegetable farms in Florida and Puerto Rico are benefiting from biological control of invasive mole crickets.

Taylor’s power law and Iwao’s patchiness regression were used to analyze spatial distribution of eggs of the Diaprepes root weevil, Diaprepes abbreviatus (L.), on silver buttonwood trees, Conocarpus erectus, during 1997 and 1998. Taylor’s power law and Iwao’s patchiness regression provided similar descriptions of variance-mean relationship for egg distribution within trees. Sample size requirements were determined. Information presented in this paper should help to improve accuracy and efficiency in sampling of the weevil eggs in the future.

Torpedograss, Panicum repens L., is an adventive, rhizomatous grass species that has become an invasive weed of terrestrial, wetland, and aquatic environments in tropical and subtropical regions worldwide. Until recently, strategies for controlling torpedograss in the USA have focused almost exclusively on mechanical and chemical methods, either alone or in combination, with varied results. A survey of the arthropods and nematodes currently associated with the plant in Lake Okeechobee, Florida, was conducted as part of a feasibility study to determine whether torpedograss is an appropriate target for a classical biological control program. Overall, approximately 4,000 arthropods and 400 nematode specimens were collected. Sweep, clipped vegetation, and soil core samples were dominated by representatives of the arthropod orders Hemiptera, Hymenoptera, Diptera, and Acari. Lesion nematodes of the genus Pratylenchus were commonly associated with the roots of torpedograss. None of the organisms collected were torpedograss specialists. Although classical biological control of torpedograss is feasible based on the extent of the infestation, economic losses, resistance to conventional controls, and the report of a potentially host specific natural enemy in India, the botanical position of this grass weed will require a formal risk assessment before proceeding with a classical biological control program.

The residual effects of several reduced-risk and conventional miticides were evaluated in strawberries (Fragaria × ananassa Duchesne) on the twospotted spider mite (TSSM), Tetranychus urticae Koch (Acari: Tetranychidae) and on 2 predatory mites, Neoseiulus californicus McGregor and Phytoseiulus persimilis Athias-Henriot (Acari: Phytoseiidae). Experiments were conducted in the laboratory and greenhouse. The greenhouse experiments also tested the direct effects of the miticides on TSSM. The efficacy of conventional and reduced-risk miticides was evaluated on strawberry leaf discs and on whole plants for control of TSSM. Furthermore, the residual effects of these miticides were evaluated on whole strawberry plants against selective predatory mites. For TSSM, 5 treatments were evaluated: a conventional miticide; fenbutatin-oxide (Vendex®) and 3 reduced-risk miticides; binfenazate (Acramite 50WP®), activated garlic extract (Repel®), sesame seed and castor oil (Wipeout®), and a water-treated control. For predatory mites, the residual effects of only Acramite® and Vendex® were evaluated. Acramite® was the most effective acaricide in reducing TSSM populations in both the laboratory and greenhouse experiments. Vendex® and Wipeout® were also effective in the laboratory, but did not cause significant reduction of TSSM in the greenhouse. Repel® was the least effective of the 4 pesticides evaluated. Neither Acramite® nor Vendex® had a significant effect on either predatory mite species. However, there appeared to be more predatory mites on the Vendex®-treated plants than on the Acramite®-treated plants. There were significantly more predatory mites of both species on the cue plants, which were inoculated with TSSM versus the non-cue plants, which were not inoculated.

Two trapping systems were compared in a study in Guatemala during the wet season, May through Dec 2001. Trap/lure combinations consisting of green or yellow-based plastic McPhail-like traps baited with a synthetic 2-component lure (putrescine and ammonium acetate) and 300 mL of propylene glycol antifreeze as a preservative were compared to the traditional glass McPhail baited with torula yeast/borax and 300 mL of water. Both systems captured several key Anastrepha species including Anastrepha ludens Loew, A. obliqua, Macquart, A. serpentina Weidemann, A. striata Schiner, A. distincta Greene, A. fraterculus Weidemann as well as Ceratitis capitata Weidemann. Additionally, 13 other Anastrepha spp. were captured with the synthetic lure. The plastic traps captured more key flies than the McPhail trap except for A. distincta where there were no significant differences between the yellow-based plastic trap and the McPhail trap and no significant differences between any trap and lure for trapping A. fraterculus. The synthetic lure lasted 10 weeks. The sex ratio was female-biased for almost all captured key species in both systems. Moreover, there were significant numbers of captured nontarget insects in all traps; however, the captured flies in those traps with the synthetic lure were not adversely affected by these insects. Propylene glycol-based antifreeze was a superior preservative when compared to borax/water.