Confronting environments with low relative humidity is one of the main challenges faced by insects with expanding distribution ranges. Anastrepha ludens (the Mexican fruit fly) has evolved to cope with the variable conditions encountered during its lifetime, which allows it to colonise a wide range of environments. However, our understanding of the mechanisms underpinning the ability of this species to confront environments with low relative humidity is incomplete. In this sense, omic approaches such as transcriptomics can be helpful for advancing our knowledge on how this species copes with desiccation stress. Considering this, in this study, we performed transcriptomic analyses to compare the molecular responses of laboratory-reared A. ludens exposed and unexposed to desiccation. Data from the transcriptome analyses indicated that the responses to desiccation are shared by both sexes. We identified the up-regulation of transcripts encoding proteins involved in lipid metabolism and membrane remodelling, as well as proteases and cuticular proteins. Our results provide a framework for understanding the response to desiccation stress in one of the most invasive fruit fly species in the world.

The Ceratitis FARQ species complex consists of four highly destructive agricultural pests of Africa, namely C. fasciventris, C. anonae, C. rosa, and C. quilicii. The members of the complex are considered very closely related and the species limits among them are rather obscure. Their economic significance and the need for developing biological methods for their control makes species identification within the complex an important issue, which has become clear that can only be addressed by multidisciplinary approaches. Chromosomes, both mitotic and polytene, can provide a useful tool for species characterization and phylogenetic inference among closely related dipteran species. In the current study, we present the mitotic karyotype and the polytene chromosomes of C. rosa and C. quilicii together with in situ hybridization data. We performed a comparative cytogenetic analysis among the above two species and C. fasciventris, the only other cytogenetically studied member of the FARQ complex, by comparing the mitotic complement and the banding pattern of the polytene chromosomes of each species to the others, as well as by studying the polytene chromosomes of hybrids between them. Our analysis revealed no detectable chromosomal rearrangements discriminating the three FARQ members studied, confirming their close phylogenetic relationships.

Methoprene, a juvenile hormone analog, is used to accelerate sexual maturation in males of species of economic importance in support to the sterile insect technique (SIT). In the SIT, mass-reared sterile males are released into the field and need to survive until they reach sexual maturation, find a wild female, mate with her and then induce female sexual refractoriness, so she will not remate with a wild counterpart. The use of methoprene shortens the time between release and copulation. However, in South American fruit flies, Anastrepha fraterculus, the ability of methoprene-treated males to inhibit female remating has been shown to be lower than wild males, when methoprene was applied by pupal immersion or topical application. Here we evaluated the possibility of incorporating methoprene into the male diet at different doses and the ability of those males to inhibit female remating, as well as the effect of methoprene on male reproductive organ size, due to the possible correlation between male accessory gland size and their content, and the role of male accessory gland proteins in female inhibition. We found that A. fraterculus males fed with methoprene in the adult protein diet at doses as high as 1% were less likely to inhibit female remating, however, at all other lower doses males had the same ability as untreated males to inhibit female remating. Males fed with methoprene had bigger male accessory glands and testes compared to methoprene-deprived males. We demonstrate that the incorporation of methoprene in adult male diets is possible in this species and potentially useful as a post-teneral, pre-release supplement at doses as low as 0.01%. Even at higher doses, the percentage of females remating after 48 h from the first copulation is sufficiently low in this species so as not compromise the efficiency of the SIT.

Agriculture is one of the major farming activities, representing 32% of the gross domestic product of Madagascar and 74.3% of the population is involved in this activity. Fruit flies of the Tephritidae family are considered as the most destructive pests for agriculture in the country, nevertheless, few data exist on host plants and distribution of those pests. In the present study, we address those questions by conducting a large survey between November 2016 and July 2018 across the six agroecological regions of Madagascar. Fruit and vegetable were sampled from 198 plant species (wild and cultivated) and represented 37,965 fruits from all regions of Madagascar. The infestation index ranged from 0.06 to 538.46 pupae/kg, the infestation percentage was up to 54.84% in some samples, 63 plant species were considered as host of Tephritidae. Twelve fruit fly species were identified, seven of which were previously described as endemic, five species could be considered as widespread (altitudinal gradients between 1 and 1634 m asl) and major pests in Madagascar: Ceratitis malgassa (23 plant species from 12 families), Neoceratitis cyanescens (16 plant species from one family), Bactrocera dorsalis (18 plant species from 12 families), Dacus demmerezi (ten plant species from one family), Dacus vertebratus (six species from one family). Those results are of importance for implementation of control measures.

Many parasitoid species discriminate already parasitized hosts, thus avoiding larval competition. However, females incur in superparasitism under certain circumstances. Superparasitism is commonly observed in the artificial rearing of the parasitoid Diachasmimorpha longicaudata, yet host discrimination has been previously suggested in this species. Here, we addressed host discrimination in virgin D. longicaudata females in a comprehensive way by means of direct and indirect methods, using Ceratitis capitata and Anastrepha fraterculus which are major fruit fly pests in South America. Direct methods relied on the description of the foraging behaviour of females in arenas with parasitized and non-parasitized host larvae. In the indirect methods, healthy larvae were offered to single females and the egg distributions were compared to a random distribution. We found that D. longicaudata was able to recognize parasitized host from both host species, taking 24 h since a first parasitization for A. fraterculus and 48 h for C. capitata. Indirect methods showed females with different behaviours for both host species: complete discrimination, non-random (with superparasitism), and random distributions. A larger percentage of females reared and tested on A. fraterculus incurred in superparasitism, probably associated with higher fecundity. In sum, we found strong evidence of host discrimination in D. longicaudata, detecting behavioural variability associated with the host species, the time since the first parasitization and the fecundity of the females.

The family Tephritidae (order: Diptera), commonly known as fruit flies, comprises a widely distributed group of agricultural pests. The tephritid pests infest multiple species of fruits and vegetables, resulting in huge crop losses. Here, we summarize the composition and diversity of tephritid gut-associated bacteria communities and host intrinsic and environmental factors that influence the microbiome structures. Diverse members of Enterobacteriaceae, most commonly Klebsiella and Enterobacter bacteria, are prevalent in fruit flies guts. Roles played by gut bacteria in host nutrition, development, physiology and resistance to insecticides and pathogens are also addressed. This review provides an overview of fruit fly microbiome structure and points to diverse roles that it can play in fly physiology and survival. It also considers potential use of this knowledge for the control of economically important fruit flies, including the sterile insect technique and cue-lure baiting.

The gut of the agricultural pest Ceratitis capitata hosts a varied community of bacteria, mainly Enterobacteriaceae, that were implicated in several processes that increase the fitness of the insect. In this study, we investigated the antagonistic activity in vitro of Klebsiella oxytoca strains isolated in the 1990s from the alimentary tract of wild medflies collected from different varieties of fruit trees at diverse localities. Assays were carried out against reference strains (representative of Gram-negative and -positive bacterial species) of the American Type Culture Collection (ATCC). Eight Klebsiella, out of 11, expressed a killing activity against Escherichia coli ATCC 23739, and Enterobacter cloacae ATCC 13047; among the eight strains, at least one showed activity against Salmonella typhimurium ATCC 23853. Genomic DNA derived from all Klebsiella strains was then subjected to PCR amplification using specific primer pairs designed from each of the four bacteriocin (KlebB, C, D, CCL) sequences found so far in Klebsiella. KlebD primer pairs were the only to produce a single product for all strains expressing the killing phenotype in vitro. One of the amplicons was cloned and sequenced; the DNA sequence shows 93% identity with a plasmid-carried colicin-D gene of a strain of Klebsiella michiganensis, and 86% identity with the sequence encoding for the klebicin D activity protein in K. oxytoca. Our work provides the first evidence that dominant symbiotic bacteria associated with wild medfly populations express a killing phenotype that may mediate inter and intraspecies competition among bacterial populations in the insect gut in vivo.

Bactrocera dorsalis (Hendel) and Ceratitis capitata (Wiedemann) are highly polyphagous fruit fly species and important pests of commercial fruit in regions of the world where they are present. In South Africa, B. dorsalis is now established in the north and northeastern parts of the country. B. dorsalis is currently absent in other parts of the country including the Western Cape Province which is an important area for the production of deciduous fruit. C. capitata is widespread in South Africa and is the dominant pest of deciduous fruit. The demographic parameters of B. dorsalis and C. capitata on four deciduous fruit types Prunus persica (L.) Batsch, Prunus domestica L., Malus domestica Borkh. and Pyrus communis L. were studied to aid in predicting the potential population establishment and growth of B. dorsalis in a deciduous fruit growing environment. All deciduous fruit types tested were suitable for population persistence of both B. dorsalis and C. capitata. Development was fastest and survival highest on nectarine for both species. B. dorsalis adults generally lived longer than those of C. capitata, irrespective of the fruit types that they developed from. B. dorsalis had a higher net reproductive rate (Ro) on all deciduous fruit tested compared to C. capitata. However, the intrinsic rate of population increase was estimated to be higher for C. capitata than for B. dorsalis on all fruit types tested primarily due to C. capitata's faster generation time. Provided abiotic conditions are optimal, B. dorsalis would be able to establish and grow in deciduous fruit growing areas.

This study aimed to evaluate the effects of toxic baits and attractants for fruit flies on the biology of its parasitoid Fopius arisanus. We tested two food-based attractants; hydrolysed corn protein (Biofruit® 3%) and sugarcane molasses (7%), their mixtures with spinosad and malathion-based insecticides, and a ready-to-use commercial bait (Success 0.02 CB®). Malathion-based lures were used as references for mortality (i.e., positive control), while negative control was honey. The formulations Biofruit® + malathion (T1), molasses + malathion (T2), and spinosad + molasses (T3) were toxic to F. arisanus, being classified as harmful (class 4). In addition, toxic baits composed of Biofruit + spinosad (T4) reduced parasitism by 97.99%, being rated as moderately harmful (class 3). Yet, Success 0.02 CB® (T7) was considered slightly toxic (class 2), causing a 64.55% reduction in parasitism. Regarding the biological parameters of F. arisanus, offspring number and parasitoid longevity were significantly reduced by using hydrolysed protein attractants when compared to the control (honey). However, sugarcane molasses improved parasitoid reproduction and longevity, as did the honey. Lastly, ingestion tests showed the major role of attractants in toxic-bait formulations against F. arisanus.

Parasitoids, released in augmentative biological control programmes, which display a rapid host-location capacity, have a higher likelihood of successfully controlling target pest species. By learning to associate sensory cues to a suitable oviposition site, might parasitoids used as biological control agents, locate hosts more rapidly, and perhaps increase the efficacity of e.g. Tephritidae fruit fly management. We studied associative learning of Fopius arisanus (Hymenoptera: Braconidae) and tested its range of learning in natural and conditional hosts and host fruits, i.e. Bactrocera dorsalis, Zeugodacus cucurbitae, Ceratitis capitata and Ceratitis cosyra (Diptera: Tephritidae) and on fruits (papaya, tomato, banana). Naïve female F. arisanus were compared with experienced wasps, which had been offered infested and non-infested fruit, and been allowed to oviposit. Preferences for olfactory cues from infested fruits were thereafter assessed in a two-choice olfactometer. Naïve and trained parasitoids preference differed in general and non-responders to infested fruits were higher among naïve parasitoids. The trained wasps preferred the fruit infested in the training more than the control fruit, for all combination, except when C. cosyra infested the fruits, hence avoidance behavioural response was observed towards the odour of the infested fruit. Fopius arisanus was capable of behaviourally respond to the learned information, e.g. associative odour learning was achieved, yet limited depending on interaction level, fruit fly and fruit combination. To create F. arisanus preference of an associated odour, it might hence be needed to ensure oviposition in perceived suitable host and host fruit, for the parasitoid learning to become favourable in a biological control setup.

The genus Rhagoletis (Diptera: Tephritidae) comprises more than 65 species distributed throughout Europe, Asia and America, including many species of high economic importance. Currently, there are three Rhagoletis species that infest fruits and nuts in Europe. The European cherry fruit fly, Rhagoletis cerasi (may have invaded Europe a long time ago from the Caucasian area of West Asia), and two invasive species (recently introduced from North America): the eastern American cherry fruit fly, R. cingulata, and the walnut husk fly, R. completa. The presence of different Rhagoletis species may enhance population dynamics and establish an unpredictable economic risk for several fruit and nut crops in Europe. Despite their excessive economic importance, little is known on population dynamics, genetics and symbiotic associations for making sound pest control decisions in terms of species-specific, environmental friendly pest control methods. To this end, the current paper (a) summarizes recently accumulated genetic and population data for the European Rhagoletis species and their association with the endosymbiont Wolbachia pipientis, and (b) explores the possibility of using the current knowledge for implementing the innovative biological control methods of sterile insect technique and incompatible insect technique.

Hypotheses about the worldwide colonization routes of the melon fly, Zeugodacus cucurbitae (Diptera: Tephritidae), are mainly based on sparse historical records. Here we aim at reconstructing the colonization history of the African continent based on an improved description of the population structure of Z. cucurbitae and approximate Bayesian analyses. Individuals of Z. cucurbitae were sampled in 17 localities from East, West and Central Africa and genotyped at 19 microsatellite markers. Bayesian analyses showed intracontinental population structuring with populations from Uganda diverging from those of Tanzania and populations from Burundi and Kenya showing traces of admixture with West African samples. Approximate Bayesian Computation provided support to the hypothesis of a single introduction Z. cucurbitae into East Africa and subsequent expansion to West Africa, each colonization event was followed by a bottleneck that promoted population divergence within Africa. Parameter estimates suggested that these events are roughly compatible with the historical records of Z. cucurbitae presence in sub-Saharan Africa (viz. 1936 in East Africa and 1999 in West Africa) and allow excluding alternative hypotheses on older or multiple introductions of Z. cucurbitae.

The sterile insect technique (SIT) is a key element for the integrated management of pest populations of the Mexican fruit fly, Anastrepha ludens, in Mexico. Its success depends on the survival of mass-reared sterile males and their ability to mate with wild females. However, colonization and mass-rearing conditions can adversely affect their ability to avoid predators. To test if colony management strategies could contribute to improve field survival abilities of mass-reared flies, we compared the survival of males exposed to the orb-weaver spider Argiope argentata. Males compared originated from three strains with different colonization strategies: (a) a colony started from field-collected wild flies (replacement), (b) a colony started by hybridizing wild males with mass-reared adapted females (hybrid) and (c) a colony started with mass-reared males selected on the basis of their survival ability and mating competitiveness in field cages (selected). Mass-reared males and wild males were used as controls. Males were exposed to spiders under laboratory cage conditions. Overall, wild males showed better survival ability than mass-reared males. Regarding the colonization approach, wild males survived better than a hybrid, replaced and selected males. We conclude that mass-rearing conditions have a strong negative effect on the ability of males to escape spiders. The colonization systems evaluated did not counter this effect. The lower survival of males from the selected colony suggests that the selection over one generation did not contribute to improve males’ predator avoidance and escape abilities and probably needs to be modified. Possible explanations for this and implications on colonization and colony management for SIT purpose are discussed.

Fruit flies in the genus Bactrocera are global, economically important pests of agricultural food crops. However, basic life history information about these pests, which is vital for designing more effective control methods, is currently lacking. Artificial diets can be used as a suitable replacement for natural host plants for rearing fruit flies under laboratory conditions, and this study reports on the two-sex life-table parameters of four Bactrocera species (Bactrocera correcta, Bactrocera dorsalis, Bactrocera cucurbitae, and Bactrocera tau) reared on a semi-artificial diet comprising corn flour, banana, sodium benzoate, yeast, sucrose, winding paper, hydrochloric acid and water. The results indicated that the larval development period of B. correcta (6.81 ± 0.65 days) was significantly longer than those of the other species. The fecundity of B. dorsalis (593.60 eggs female−1) was highest among the four species. There were no differences in intrinsic rate of increase (r) and finite rate of increase (λ) among the four species. The gross reproductive rate (GRR) and net reproductive rate (R0) of B. dorsalis were higher than those of the other species, and the mean generation time (T) of B. cucurbitae (42.08 ± 1.21 h) was longer than that of the other species. We conclude that the semi-artificial diet was most suitable for rearing B. dorsalis, due to its shorter development time and higher fecundity. These results will be useful for future studies of fruit fly management.

Dormancy can be defined as a state of suppressed development allowing insects to cope with adverse conditions and plant phenology. Among specialized herbivorous insects exploiting seasonal resources, diapause frequently evolves as a strategy to adjust to predictable plant seasonal cycles. To cope with acyclic and unpredictable climatic events, it has been found for some insects that a proportion of the population undergoes prolonged dormancy. We compared the response of three species in the Rhagoletis cingulata species group exploiting plants differing in fruiting phenology from environments varying in frequency and timing of acyclic climatic catastrophic events (frost during flowering and fruit set) and varying also in the time of the onset of the rainy season. Small proportions (<2%) of Central and Northeastern Mexico R. cingulata and Rhagoletis turpiniae, and a few associated adult parasitoids, emerged without becoming dormant. Pupae exposed to 5°C for different periods of time (0–16 weeks) became dormant and emerged as adults in proportions <30%, compared with other studied species in the group from the USA (up to 80%). Large proportions (19–55%) of exposed pupae entered prolonged dormancy (>10 months), and large proportions of pupae died without emerging as adults. The number of days elapsed from the end of artificial winter and adult eclosion was longer for R. cingulata exploiting late fruiting Prunus serotina in Northeastern Mexico than for flies recovered from earlier fruiting plants in the central Altiplano. Rhagoletis turpiniae and northeastern R. cingulata pupae suffered high proportions of parasitism. Large proportions of R. cingulata from central Mexico engaging in prolonged dormancy may be explained by the fact that flowering and fruit set for its host, P. serotina var capuli, driven by the timing of maximum precipitation, matches a period of highest probability of frost often resulting in large areas with fruitless trees at unpredictable time intervals. As a consequence of differences in host plant fruiting phenology, central and northeastern Mexican R. cingulata were found to be allochronically isolated. Prolonged dormancy may have resulted in escape from parasitism.

A survey was conducted at 88 sites in Montana in 1984 and 1985 to determine the distribution of Urophora affinis Frauenfeld and U. quadrifasciata (Meigen), two seed head flies released on spotted knapweed (Centaurea maculosa Lam. # CENMA) in North America. U. affinis, released in Montana during 1973 to 1977, was found at 40 of the 88 sites. The fly was concentrated within a short radius (about 5 to 8 km) of release sites, even at sites having well-established populations. U. quadrifasciata, released in British Columbia in 1972, but not in the United States, was found at 84 sites. The data suggest that the fly entered the state in the northwest corner and dispersed in a southeasterly direction. There was a strong positive relationship between mean number of galls per seed head and percent infestation for both fly species.

Olfaction is a key sense routing foraging behaviour in parasitoids. Preferences for food, mate and host stimuli can be innate in parasitic wasps. Alternatively, learning-mediated mechanisms play a crucial role. Females of the braconid parasitoid Psyttalia concolor exploit olfactory cues arising from tephritid hosts and related microhabitats. However, little is known on the olfactory stimuli routing males searching for mates. In this study, we focused on the attractiveness of Bactrocera oleae-induced olive volatiles towards P. concolor males. Furthermore, we evaluated learning occurrence in virgin males, when trained for selected unattractive volatile organic compounds (VOCs) associated with mate rewards. (E)-β-Ocimene, α-pinene and limonene attracted virgin males in Y-tube bioassays. Unattractive VOCs evoked positive chemotaxis after associative learning training. P. concolor males exposed to VOCs during a successful or unsuccessful mating, showed short-term preference for these VOCs (<1 h). However, memory consolidation was strictly dependent on reward value. Indeed, males experiencing a successful mating showed a fast consolidation into protein dependent long-term memory, appearing after 24 h. On the other hand, males experiencing a less valuable training experience (i.e. unsuccessful courtship), did not show consolidated memory after 24 h. Overall, our findings suggest that P. concolor virgin males may exploit VOCs from the host microhabitat to boost their mate searching activity, thus their reproductive success. However, since learning is a costly process, P. concolor males retained durable memories just in presence of a valuable reward, thus avoiding maladaptive behaviours.

Dormancy has been thoroughly studied for several species of economic importance in the genus Rhagoletis in temperate areas of North America and Europe. Much less is known on life history regulation for species inhabiting high-elevation areas in the subtropics at the southern extreme of their geographical range. Host plant phenology has been found to play a key role in generating allochronic isolation among sibling species and host races of Rhagoletis in the course of sympatric speciation, and has important implications for pest management. We compare the effect of winter length on survival to adult eclosion and dormancy duration among four species of Rhagoletis (three of them sympatric) exploiting hosts with different fruiting phenology in subtropical isolated highlands. Survival and duration of dormancy was found to be different among the four species. At 24°C, a very small proportion (<1%) of R. pomonella, R. turpiniae and R. zoqui completed development without becoming dormant, while in the case of R. solanophaga the majority of the population emerged after development within 40 days of pupation. Also, a large proportion of braconid parasitoids infesting Rhagoletis eggs and larvae emerged as adults without becoming dormant. Greatest survival after artificial winter was obtained for R. pomonella (50–60%) and R. zoqui (30%) after only four weeks at 5°C (a third of the time reported for studies on northern R. pomonella), while R. turpiniae, under identical environmental conditions experienced low adult emergence, and highest survival (11%) was recorded for flies exposed to 5°C during 10 and 12 weeks. For R. pomonella, there was a strong positive relationship between winter length and time to post-winter adult eclosion that was not observed for R. zoqui. In sum, for R. pomonella, mild winters in highland subtropical areas appear to select for flies better able to withstand longer periods of warm temperature before winter than flies exploiting late fruiting hosts and inhabiting northern latitudes. In the case of R. turpiniae and R. zoqui environmental cues such as fluctuations in humidity and/or different temperature thresholds (5°C) may play a more important role than winter length in life history regulation. Continuous host availability for R. solanophaga appears to have selected for non-diapausing flies. From an applied perspective our results are useful for handling flies in the laboratory to conduct research and suggest that non-diapausing strains of flies and parasitoids may be selected for SIT and innundative biological control programs.

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.

Anastepha ludens (Diptera: Tephritidae) is a highly polyphagous fruit fly that is able to develop in a wide range of hosts. Understanding the limits of this pest's host range could provide valuable information for pest management and plant breeding for pest resistance. Previous studies have shown that guavas (Psidium guajava (Myrtaceae) L.), are not attacked under natural conditions by A. ludens. To understand this phenomenon, guavas were exposed to natural infestation by A. ludens and to other fruit fly species that infest guavas in nature (Anastrepha striata Schiner, Anastepha fraterculus (Wiedemann), Anastepha obliqua (Macquart)). Once the susceptible phenological stage of guavas was determined, fruit infestation levels were compared between A. ludens and A. striata. Choice and non-choice tests were performed under field-cage conditions. Under field conditions, guavas were susceptible to A. striata and A. fraterculus attack all the way from when fruit was undeveloped to when fruit began to ripen. No infestation by A. ludens was recorded under natural conditions. Similar results were obtained when forced exposures were performed, indicating that unripe guavas were preferred by A. striata over ripe fruit, and that infestation rates were higher at early fruit maturity stages. Under forced oviposition conditions, A. ludens larvae were unable to develop in unripe guavas but did so in fully ripe fruit. However, A. ludens fitness parameters were dramatically affected, exhibiting reduced survival and reduced pupal weight compared to conspecifics that developed in a natural host, grapefruit. We confirm that P. guajava should not be treated as a natural host of this pestiferous species, and suggest that both behavioral aspects and the fact that larvae are unable to adequately develop in this fruit, indeed represent clear limits to A. ludens's broad host range.