Host availability affects the interaction between pupal parasitoid Coptera haywardi (Hymenoptera: Diiapridae) and larval-pupal parasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae).

The use of multiple species in biological control programmes is controversial when interactions among them are not fully understood. We determined the response of the pupal parasitoid Coptera haywardi (Oglobin) to different availability of Anastrepha ludens (Loew) pupae previously parasitized or not by larval-pupal Diachasmimorpha longicaudata (Ashmead). The two types of pupae were exposed at different ages and proportions to different numbers of C. haywardi females for 48 h. The performance of C. haywardi adults emerging from parasitized and unparasitized pupae was measured. Coptera haywardi prefers to attack unparasitized A. ludens pupae rather than pupae parasitized by D. longicaudata. However, when the availability of unparasitized pupae was low or the number of foraging females was high, C. haywardi competed against early immature stages of the D. longicaudata, or hyperparasitized, feeding directly on the advanced-immature developmental stages of the early acting species. Adults of C. haywardi emerging as hyperparasitoids were no different in size, fecundity and longevity from those emerging as primary parasitoids. Our data suggest that simultaneous use of these species in augmentative biological control projects may be feasible but should be carefully planned in order to avoid any detrimental effect of its interaction.

Phenotypic plasticity in clutch size regulation among populations of a potential invasive fruit fly from environments that vary in host heterogeneity and isolation.

Phenotypic plasticity is thought to evolve in response to environmental unpredictability and can shield genotypes from selection. However, selection can also act on plastic traits. Egg-laying behaviour, including clutch size regulation, is a plastic behavioural trait among tephritid fruit flies. We compared plasticity in clutch size regulation among females of Anastrepha ludens populations stemming from environments that differed in the degree of predictability in egg-laying opportunities. Clutch size regulation in response to hosts of different sizes was compared among flies from (a) a wild, highly isolated population, (b) a wild population that switches seasonally from a small wild host fruit that varies greatly in abundance to an abundant large-sized commercial host, and (c) a laboratory population. Flies from all three populations adjusted clutch number and size according to host size. However, flies from the heterogeneous wild environment were more plastic in adjusting clutch size than flies from agricultural settings that also laid fewer eggs; yet both populations were more plastic in adjusting clutch size in line with host size when compared with laboratory females. When wild and orchard females encountered the largest host, clutch size was extremely variable and egg regulation did not follow the same trend. Heterogeneity in host availability in space and time appears to be as important as seasonal variation in host size in maintaining plastic clutch size regulation behaviour. In stable environments, there was a clear reduction in the plasticity of these traits.

Antipredator behavior of the new mass-reared unisexual strain of the Mexican Fruit Fly.

Tephritid pests controlled through the sterile insect technique (SIT) are mass-reared and subsequently released in affected areas. Several quality parameters are currently used to test adults, but none take into account interactions with a predator. When sterile males are released in the field, they will need to avoid predators until they reach sexual maturity and survive long enough to mate with wild females. Spiders are one of the most common predators that flies may encounter in release sites. In this study, we evaluated the antipredator behavior of a mass-reared sterile unisexual strain ('Tapachula-7') of the Mexican fruit fly Anastrepha ludens (Diptera: Tephritidae) against their spider predators. We sampled spiders in citrus trees to determine which families could be more common. We established the baseline activity rates of sterile Tapachula-7 (Tap-7) flies in comparison with wild flies. We also tested the behavior of the fertile and sterile bisexual strain and wild flies against hunting spiders (Family Salticidae) and orb building spiders (Family Tetragnathidae). We recorded 18 spider families, with Salticidae being the most dominant. Tap-7 flies diminished their activity in comparison with wild males at 1800 h but showed similar activity levels earlier in the day. When exposed to orb-web spiders (Leucauge venusta), Tap-7, fertile and sterile males from the bisexual strain had similar rates of survival, but Tap-7 males showed lower survival than wild males. Against hunting spiders (Phidippus audax), wild males had higher probability of defensive wing displays, but there was no difference in spider attack rates. In general, sterile Tap -7 males performed as well as males from the bisexual strain, although they had lower survival than wild males. This could be due to either mass-rearing and/or irradiation effects. We recommend the use of the defensive wing display behavior as a quality parameter and propose a rapid and effective method to evaluate fly activity. The efficiency of SIT will be improved if released sterile males have the same antipredator repertoire as their wild counterparts.

The effect of irradiation and mass rearing on the anti-predator behaviour of the Mexican fruit fly, Anastrepha ludens (Diptera: Tephritidae).

Fruit flies (Diptera: Tephritidae) are major pests worldwide. The sterile insect technique, where millions of flies are reared, sterilized by irradiation and then released, is one of the most successful and ecologically friendly methods of controlling populations of these pests. The mating behaviour of irradiated and non-irradiated flies has been compared in earlier studies, but there has been little attention paid to the anti-predator behaviour of mass-reared flies, especially with respect to wild flies. Tephritid flies perform a supination display to their jumping spider predators in order to deter attacks. In this study, we evaluated the possibility of using this display to determine the anti-predator capabilities of mass-reared irradiated, non-irradiated flies, and wild flies. We used an arena setup and observed bouts between jumping spiders (Phidippus audax Hentz) and male Mexican fruit flies (Anastrepha ludens Loew). We show that although all flies performed a supination display to their predator, wild flies were more likely to perform a display and were significantly more successful in avoiding attack than mass-reared flies. We suggest that this interaction can be used to develop a rapid realistic method of quality control in evaluating anti-predator abilities of mass-reared fruit flies.

Susceptibility of 15 mango (Sapindales: Anacardiaceae) cultivars to the attack by Anastrepha ludens and Anastrepha obliqua (Diptera: Tephritidae) and the role of underdeveloped fruit as pest reservoirs: management implications.

We evaluated the susceptibility of 15 mango cultivars to the attack of Anastrepha ludens (Loew) and Anastrepha obliqua (Macquart) (Diptera: Tephritidae), the main tephritid pests of this crop in Mexico. In a field experiment, bagged fruit-bearing branches were exposed to gravid females of both fly species. Infestation rates, developmental time, adult eclosion, and F1 adult longevity, fecundity, and fertility were recorded, ranking cultivars in terms of susceptibility to fly attack and development. We also compared the volatile profile in selected resistant and susceptible cultivars in search of possible correlations. In a second experiment, clutch size for A. ludens was determined in each cultivar. Infestation rates, developmental time, and F1 demographic parameters varied sharply among cultivars and between fly species for bagged fruit. Cultivars 'Vishi,' '74-82,' and 'Brooks' were most susceptible to A. ludens infestation while "Tommy,' 'Sensation,' and 'Ataulfo "niño"' (parthenocarpic fruit) were most susceptible to A. obliqua infestation. 'Edward,' 'Kent,' 'Brooks late,' 'Palmer, and 'Ataulfo' exhibited tolerance to attack of both fly species. Fruit of susceptible and resistant cultivars exhibited unique volatile profiles. Fly development and F1 adult demographic parameters varied significantly among cultivars. A. ludens females laid larger clutches in larger and harder fruit. We highlight the important role of Ataulfo "niño" as pest reservoir if fruit is left unharvested on trees. We discuss the possible use of highly resistant cultivars as trap crops or egg sinks.

The Sterile Insect Technique: Success and Perspectives in the Neotropics.

The sterile insect technique (SIT), an environmentally friendly means of control, is currently used against plant, animal, and human pests under the area-wide integrated pest management. It consists in the mass production, sterilization, and release of insects in an affected area where sterile males mate with wild females leading to no reproduction. Here, we review SIT in the Neotropics and focus on particular recent successful cases of eradication of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), as well as effective programs used against the Mexican fruit fly Anastrepha ludens (Loew), the New World screwworm fly Cochliomyia hominivorax (Coquerel)), and the Cactus moth Cactoblastis cactorum (Berg). We examine when SIT does not work and innovations that have made SIT more efficient and also highlight complimentary techniques that can be used in conjunction. We address potential candidate species that could be controlled through SIT, for example Philornis downsi Dodge & Aitken. Finally, we consider the impact of climate change in the context of the use of the SIT against these pests. Given the recent dramatic decline in insect biodiversity, investing in environmentally friendly means of pest control should be a priority. We conclude that SIT should be promoted in the region, and leadership and political will is needed for continued success of SIT in the Neotropics.

Effect of cold storage on larval and adult Anastrepha ludens (Diptera: Tephritidae) viability in commercially ripe, artificially infested Persea americana 'Hass'.

Commercially ripe 'Hass' avocados, Persea americana Mill, artificially exposed to wild Anastrepha ludens (Loew) (Diptera: Tephritidae) females 24 h after harvest were placed in a cold storage facility to determine the effect of low temperature on larval survival and adult viability. Fruit were left for 3, 6, 9, and 12 d in a cold room at 5 degrees C followed by a 20-25-d period at ambient temperature to allow for larval development and pupation. Hass avocados and grapefruit, Citrus paradisi Macfadyen, maintained at ambient temperature served as controls. Overall, only 0.23% of the Hass avocados and 19.30% of the grapefruit were infested. The number of infested fruit increased with decreasing exposure time to cold. Puparia from cold-treated Hass avocados were significantly smaller than those stemming from cold-treated grapefruit. Hass avocados exposed for 12 d to 5 degrees C yielded no puparia, and those exposed for 6 and 9 d yielded 22 and two puparia, respectively, but no adults. Although Hass avocados exposed to cold temperature for 3 d yielded adults that reached sexual maturity (N = 16), females laid inviable eggs. Grapefruit exposed to cold for 12 d yielded normal-sized puparia (but no adults), whereas those exposed over 9 d yielded females able to lay viable eggs. We conclude that exposing fruit to cold storage after packing and during transport represents an effective risk-mitigating procedure in the highly improbable event that a gravid A. ludens female might lay eggs in a commercially ripe Hass avocado that had been left unprotected in a packinghouse.

Random mating among Anastrepha ludens (Diptera: Tephritidae) adults of geographically distant and ecologically distinct populations in Mexico.

The Mexican fruit fly Anastrepha ludens (Loew) (Diptera: Tephritidae) is a polyphagous pestiferous insect with a geographical range encompassing highly variable environmental conditions. Considering that cryptic species have been recently found among South American representatives of the same taxonomic group as A. ludens, we tested whether or not some populations of A. ludens have evolved assortative mating as an isolating mechanism that maintains intrapopulation genetic differences and behavioral adaptations to local conditions. Males and females stemming from widely separated locations with similar environmental conditions and males and females stemming from populations within individual-flight range, but collected in different hosts (a native and an exotic one), mated randomly amongst themselves when placed in a field cage. Despite the fact that sibling males and females from two distinct populations also mated randomly amongst themselves, siblings engaged in significantly longer copulations than non-siblings, indicating that perhaps adults discriminated mates with similar genetic compositions. Our results have important practical implications as A. ludens is the most devastating pest of citrus in Mexico and Central America, and large-scale releases of sterile flies are used to control it.

Application of feces extracts and synthetic analogues of the host marking pheromone of Anastrepha ludens significantly reduces fruit infestation by A. obliqua in tropical plum and mango backyard orchards.

We determined the efficacy of three potential oviposition deterrents in reducing fruit infestation by Anastrepha obliqua in tropical plum and mango orchards. These were: (1) Extracts of feces of Mexican fruit fly, Anastrepha ludens, known to contain the A. ludens host marking pheromone (HMP) and (2) two fully synthetic simplified analogues of the naturally occurring compound, which we have named desmethyl A. ludens HMP (DM-HMP) and Anastrephamide. Two applications of feces extracts 2 or 3 wk before fruit color break reduced A. obliqua infestation in plums by 94.1, 75.9, and 72% when measured 8, 14, and 25 d, respectively, after application. The natural A. ludens-HMP containing extract retained its effectiveness despite considerable rainfall (112.5 mm) and high A. obliqua populations. The synthetic desmethyl HMP derivative (DM-HMP) also reduced infestation in plums by 53.3 and 58.7% when measured, 18 and 26 d, respectively, after application. Finally, applications of Anastrephamide resulted in fruit loss cut by half and an 80% reduction in numbers of fly larvae per fruit. Our results confirm previous findings indicating that there is interspecific cross-recognition of the HMP in two of the most pestiferous Anastrepha species and open the door for the development of a highly selective, biorational Anastrepha management scheme.

Behavioral Responses and Pupa Development Patterns After Hypoxia or Anoxia in a Desiccation-Resistant Anastrepha ludens Strain.

The aim of this study was to experimentally test for adult cross-resistance to hypoxia or anoxia in a desiccation-resistant population of Anastrepha ludens Loew. We compared desiccation resistant flies with unselected (control) flies by measuring the effect of pre-emergence hypoxia on some fitness parameters (emergence, flight ability, copulation success, latency to copulation, copulation duration, ovary size). Anoxia effects were determined using eye color changes during pupa development and fly emergence after re-oxygenation. Both strains were negatively affected in all measured parameters when exposed to hypoxia for more than 48 h. However, after hypoxia, control flies showed, in general, shorter latency to mate and longer copula duration than desiccation-resistant flies. Anoxia-induced arrest of pupa development, whereas returning to normoxia conditions induced resumption of development. Anoxia period length (longer than 72 h) increased mortality to 100% in the control line, whereas the desiccation-resistant line survived even at 120 h of anoxia. Thus, pre-release hypoxia must not exceed 24 h in order to maintain insect quality independently of fly type.

When Less Is More: Sex Ratios for the Mass-Rearing of Anastrepha ludens (Diptera: Tephritidae).

Anastrepha ludens (Loew) is one of the most important pests of citrus and mango crops in Mexico. A method used to control this pest is the sterile insect technique, which consists in the mass production, irradiation, and release of insects in affected areas. The production of insects begins with the establishment of colonies to produce eggs, which must be highly fertile to ensure an adequate production of larvae. However, female fecundity and fertility can be affected by adult density and sex ratio, thus an optimal sex ratio in mass-rearing cages must be used. The genetic sexing strain of A. ludens (Tapachula-7) allows the identification of the sex at the pupal stage, making it possible to establish rearing cages with different sex ratios. We determined if different sex ratios have an effect on egg production. Two sex ratios (4♀: 1♂ and 1♀: 1♂) were compared. Fecundity, fertility and survival at different ages were also determined. Higher fertility and fecundity per female were observed at a ratio of 4:1. However, females with higher fecundity had reduced survival probabilities. In conclusion, maintaining colonies with a lower proportion of males in cages ensures a greater fecundity and fertility. Further research is necessary to understand whether results can be attributed to lower male harassment in cages.

Effects of size, sex and teneral resources on the resistance to hydric stress in the tephritid fruit fly Anastrepha ludens.

Water availability is recognized as one of the most important factors in the distribution and activity of terrestrial organisms. In the case of insects, hydric stress imposes a major challenge for survival because of the small surface-area-to-volume ratio they exhibit. In general, stress resistance is expected to co-vary positively with size; however, this pattern can become obscured in insects that exhibit sexual size dimorphism, as sexes differ in size and/or shape and have dissimilar resource allocations. In the present study, we use an allometric-based approach to (i) assess the desiccation and starvation stress resistance of teneral Anastrepha ludens flies, (ii) disentangle the relationships between resistance, size and sex and (iii) examine the adult fly body differences in water and lipid contents before and after exposure to stress. After controlling for sexual size dimorphism, an allometric increase of resistance with overall size was observed for all stress-based treatments. The scaling exponents that define the proportion of increase resistance varied with size traits and with type and degree of hydric stress. In this allometric relationship, and also in the relationships between mass and wing length and between size and teneral resources, the sexes maintained similar scaling exponents but differed in the intercepts. Males were more resistant to stress than females; this competitive advantage is probably linked to greater amounts of teneral lipids and more water use during stress.

Oviposition deterrents for the Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae) from fly faeces extracts.

After oviposition, females of the Mediterranean fruit fly Ceratitis capitata Wiedemann deposit a host-marking pheromone on the fruit surface that deters oviposition by conspecifics. Methanolic extracts of fruit fly faeces elicit a similar deterrent effect. The results of laboratory and field experiments using raw methanolic extracts of C. capitata faeces as an oviposition deterrent are reported. Laboratory bioassays revealed a significant positive relationship between concentration of faeces and the inhibition of oviposition responses by C. capitata. Treatment of halves of coffee bushes with methanolic extracts containing 0.1, 1.0 and 10 mg faeces ml(-1) resulted in a significant reduction of infestation only at the highest concentration (P=0.03). Treatment of blocks of coffee bushes with an extract of 10 mg faeces ml(-1) resulted in an 84% reduction in infestation by C. capitata in sprayed plants and a 56% reduction in adjacent untreated coffee bushes surrounding treated plots, probably due to the deterrent effect of host-marking pheromone on fly oviposition. We conclude that faeces contain oviposition deterrent substances that effectively reduce fruit infestations by C. capitata, suggesting a clear potential for the use of this infochemical in integrated management programmes targeted at this pest.

Effects of age, diet, female density, and the host resource on egg load in Anastrepha ludens and Anastrepha obliqua (Diptera: Tephritidae).

Oocyte counts, used as a measure of egg load, were compared among three different age groups (15, 30 and 45 days) of two polyphagous species of tephritid fruit flies, Anastrepha ludens and Anastrepha obliqua, which were exposed to varying conditions of diet (sucrose vs sucrose and protein), availability of oviposition substrate (present vs absent), adult female density (1, 2 and 4 females/cage), and semiochemical context (presence vs absence of male pheromones and fruit volatiles). In both species, oocyte counts were higher in older females and for females fed sucrose and protein than for females fed sucrose only. The presence of artificial oviposition substrates influenced oocyte counts in A. obliqua, but not in A. ludens. Female density influenced oocyte counts in both species. Females maintained in groups had higher egg loads than isolated females. Finally, preliminary evidence suggests that semiochemical context influenced oocyte counts. Counts were highest for females in a room containing both fruit volatiles and male pheromone, lowest for females in a room containing neither volatiles nor pheromone, and intermediate for females in rooms containing either volatiles or pheromone but not both. Our results suggest that egg load is influenced by environmental factors in different ways in these two species. Egg load in A. obliqua, a species whose host fruits are highly ephemeral, is responsive to access to the host resource. By contrast, in A. ludens, a species infesting less ephemeral fruit, female density and age played a more important role than host stimuli. The role of ovarian maturation and oviposition in mediating these effects, as well as implications for mass rearing and pest management, are discussed.