Controlling Aedes aegypti populations through traditional methods is increasingly difficult due to the development of insecticide resistance and their use of cryptic breeding habitats. The sterile insect technique has emerged as an effective tool for integrated vector management. Still, its success depends on the ability to mass-rear large numbers of high-quality mosquitoes. Choosing an appropriate larval diet is crucial for scalable mass-rearing, as it directly influences mosquito development, survival, and overall production efficiency. This study compared the effects of 3 larval diets: 1) TetraMin® Tropical Flakes, 2) Ziegler® Tropical Pro-Start45 Meal, and 3) bovine liver powder delivered in cellulose capsules, on the growth and performance of male Ae. aegypti under simulated mass-rearing conditions. Ziegler-reared mosquitoes had significantly larger pupal and adult sizes than those reared on Tetramin or liver powder. The Tetramin diet produced smaller adults, but with longevity comparable to that of the Ziegler diet, whereas the liver powder diet resulted in mosquitoes of similar size to those of the Tetramin diet, but with reduced longevity. While all 3 diets demonstrated viability for mass-rearing, their suitability depends on program-specific goals and constraints. The liver powder diet offered a good balance of biological performance and operational efficiency, but at a substantially higher economic cost. With further optimization of feeding regimens, the Ziegler diet shows the most significant potential to deliver high biological quality at the lowest price, making it a strong candidate for scalable mass-rearing programs.

Climate change is driving the rapid range expansion of Aedes aegypti into temperate regions, presenting novel seasonal cues that can affect seasonal plasticity. Seasonal plasticity in Ae. aegypti is well studied in eggs but remains understudied in adults. Here, we investigated the effects of photoperiod and temperature on thermal plasticity, measured by the ability to cold acclimate, and other dormancy-related traits in female Ae. aegypti adults. Cold acclimation was done under a short-day (SD) and long-day (LD) photoperiod. LD photoperiod was introduced at three life stages: parental generation, egg state and adult acclimation, to determine if timing of LD photoperiod introduction affected thermal plasticity. LD photoperiod had a significant influence on both cold tolerance metrics assessed, chill coma onset temperature and survival after a cold stress. Introduction of LD photoperiod reduced thermal plasticity, with the magnitude of change in plasticity depending on when LD was introduced. At warm temperatures, LD photoperiod modified the rhythmic expression profile of timeless, one of the circadian clock genes analysed in this study, but not the other, period. In contrast, cold acclimation abolished the cyclical expression of both clock genes. Cold acclimation under both photoperiods suppressed blood-feeding behaviour, which resumed upon warming, but cold acclimation did not change relative transcript abundance of dormancy-implicated genes. This study presents the first evidence of an adult photosensitive seasonal phenotype in Ae. aegypti, with improved cold tolerance and reproductive quiescence. These findings contribute to a deeper understanding of how environmental cues may facilitate the continued expansion of this important disease vector into temperate environments.

Variable thermal environments may have both detrimental and beneficial effects. For example, extreme temperatures may challenge homeostasis and inflict tissue damage, but may also induce acclimation that improves stress resilience. Hormetic models provide a framework to understand dosage-dependent, contrasting beneficial and detrimental effects from physiological and ecological perspectives. We used a hormetic framework and associated quantitative models to investigate how a range of relatively cold, pre-exposure temperatures influence survival and fertility following cold shock at a more extreme cold temperature in the fruit fly Drosophila melanogaster. Cold pre-exposure can induce a protective rapid cold hardening (RCH) response, fail to stimulate a response, or cause direct cold injury. We found a plateau-shaped relationship between pre-exposure temperature and female survival resilience, where survival following a cold shock remained high across a range of temperatures, with sharp transitions at higher and lower temperatures. Bayesian fitting of a bi-logistic model highlights these transitions at temperature thresholds that govern processes mediating both acclimation and cold injury. In contrast to survival, female fertility resilience exhibited a muted response to pre-exposure temperature in the presence and absence of post-stress mating opportunities. Overall, a range of pre-exposure temperatures allowed low but successful reproduction following cold shock. High survival but low fertility resilience is consistent with (1) differential impacts of cold on somatic and reproductive tissues and (2) a growing body of literature suggesting that the thermal sensitivity of fertility may be more limiting than survival for population persistence in variable and changing climates.

The sterile insect technique (SIT) is increasingly used as an alternative or supplemental tool in the integrated mosquito management toolbox to protect human health worldwide. SIT programs targeting mosquitoes such as Aedes aegypti (L.) have generally used high-activity isotopes of 60Cobalt or 137Caesium to sterilize males, however, these gamma irradiators pose substantial security challenges and are becoming more difficult and expensive to obtain and maintain. One practical alternative is using commercially available low-energy X-ray irradiators. In this study, we compared the efficacy of a low-energy X-ray irradiator and a traditional gamma irradiator for achieving male sterility in both male pupae and adults of the mosquito Ae. aegypti Linn. We found that both irradiators performed equivalently with 99% sterility achieved at ~50 Gy when male pupae were irradiated and ~70 Gy when male adults were irradiated. There were no differences in lifespan or male mating competitiveness at the sterilizing dose between the 2 irradiation platforms. Taken together, our results suggest that with careful dosimetry X-ray irradiators can effectively be used as an alternative to gamma irradiators in SIT programs targeting Ae. aegypti.

Florida mosquito control districts are increasingly confronted with severe Culicoides biting midge problems in coastal areas. Yet, there is no clear guidance for integrating Culicoides management into mosquito-focused operations. This study describes population abundance and distribution trends for the biting midge Culicoides furens on a residential island in Cedar Key, Florida. We use multi-year adult trapping data to help develop strategies that may be used by mosquito control districts to target C. furens populations where they are nuisance pests. Trap data from 2005 and 2007 identified seasonal peaks, high spatial heterogeneity, and substantial year-to-year variation, with an 88.3% reduction in trap captures between 2005 and 2007. These findings provide a foundation for integrated Culicoides management strategies where legal mandates, emerging pathogen risks, and taxpayer-driven nuisance complaints may justify expanded Culicoides control activities by Florida’s Mosquito Control Districts.

Control of Aedes aegypti populations remains a major challenge due to multiple factors, including widespread development of insecticide resistance, daytime biting preference, and cryptic breeding and resting sites. Sterile Insect Technique (SIT) has emerged as a promising complementary tool within integrated vector management programs, demonstrating promising results. The mosquito control program in St. Johns County, Florida, has been developing an in-house SIT program following the phased conditional approach recommended by the World Health Organization and International Atomic Energy Agency. This study aimed to determine the minimum X-ray dose required to induce ≥99% sterility in adult male Ae. aegypti of the local strain while maintaining high sterile male quality. A dose-response experiment was conducted, followed by assessments of survival, longevity and mating competitiveness. An absorbed X-ray dose of 47.13 ± 2.09 Gy (± SD) achieved 99% sterility without significantly affecting the survival and longevity of adult males. Fried’s competitiveness index estimated for males irradiated at a target dose of 50 Gy (mean ± SD: 1.12 ± 0.12) indicated that the irradiated males were as competitive as unirradiated males. A 5:1 irradiated: unirradiated ratio achieved an 84% induced sterility, which was not significantly different at the 10:1 ratio. No significant differences in competitiveness were observed among 1:1, 5:1 and 10:1 ratios. Based on these results, a 50 Gy dose of X-rays was selected as the operational dose for adult males of the local Ae. aegypti strain.

Aedes aegypti, an aggressive nuisance biter and the primary vector for numerous arboviruses, such as chikungunya, dengue, and Zika, presents significant control challenges due to its ability to thrive in urban environments, escapes insecticide treatment by using cryptic resting and oviposition sites, and development of resistance to chemical mosquito control products being used routinely. From 2020 to 2022, the Lee County Mosquito Control District (LCMCD) employed the Sterile Insect Technique (SIT) to test a new approach in its integrated mosquito management toolbox, targeting the population suppression of Ae. aegypti on Captiva Island, Florida. Over 24.1 million sterile males were released across three phases, covering up to 142 hectares. The study demonstrated a population reduction of up to 79% in wild adults and a 59% decline in egg densities in the primary intervention area. While population control was successful, an influx of wild females from untreated zones posed ongoing challenges to complete suppression in areas close to non-intervention areas. This supported a need for an area-wide integrated pest management (AW-IPM) approach. These results underscore SIT's potential as a critical tool in integrated mosquito management strategies and emphasize practical application.

The sterile insect technique (SIT) is a highly effective biologically-based method for the suppression of many insect pest populations. SIT efficacy could be improved by methods of male sterilization that avoid the use of irradiation that can result in diminished fitness and mating competitiveness. Alternative sterilization methods include conditional disruption of genes for male fertility, or using their sperm-specific promoters to drive the expression of genes for lethal effectors. Testing has begun for the testis-specific β2-tubulin gene, though additional male fertility genes are required for redundancy or replacement, and for species where the β2-tubulin isoform does not exist or is not testis-specific. Here we had the goal of identifying and characterizing the sequence and transcriptional expression of two genes in the caribfly, Anastrepha suspensa, that are cognates of D. melanogaster spermatocyte-specific male fertility genes. In Drosophila, wampa encodes a coiled-coil dynein subunit required for axonemal assembly essential to microtubule-based sperm motility, while Prosα6T is a proteasome subunit gene required for spermatid individualization and nuclear maturation. In A. suspensa a cognate to wampa exhibited testis-specific transcript expression, which was minimal in both male and female body tissue. A Prosα6T cognate was not apparent in A. suspensa, but its constitutive isoform, Prosα6, expresses in male testes, but also in male and female body tissue. Thus, for A. suspensa, wampa remains a strong candidate gene for male sterility strategies for SIT including a direct target for gene-editing knockout and use of its promoter for testis-specific toxicity or cell death in conditional expression systems.

Acalitus simplex is an erinose eriophyoid mite that feeds exclusively on plants in the genus Ruellia, which are frequently sold as ornamentals for landscape plantings. This study presents a comprehensive examination of A. simplex collected from Anguilla, Brazil, Dominican Republic, Florida, and Hawaii through sequencing of the mitochondrial cytochrome oxidase I gene (COI), coupled with morphological examination by several microscopy techniques. The collected A. simplex specimens were similar in morphology across geographic locations and populations clustered together in a phylogenetic analysis based on COI sequences. In addition to Ruellia simplex, new host plant associations are reported in Florida, including R. blechum, R. squarrosa and R. caroliniensis. We defined the different erineum stages, determined the number of days after infestation on an uninfested plant required for erinea development, and estimated population densities in each erineum stage. Four erineum stages were observed on R. simplex - hyaline (stage 1), white (stage 2), purple (stage 3) and beige (stage 4). At 25 ± 2 oC, 12:12/L: D and 50 ± 10

Nature has already solved many challenges that synthetic biology seeks to address. At the same time, high-throughput platforms and artificial intelligence-driven design tools are redefining the landscape of synthetic biology. By integrating evolutionary insights with new enabling technologies, we are poised to move beyond modifying existing traits to engineering entirely new cellular functions. This series of vignettes explores how phenotypic engineering, inspired by nature's most extreme adaptations, represents the next frontier in synthetic biology. We first make a case for the importance of continuing to explore nature by examining how foundational discoveries in molecular biology, motivated by curiosity to understand how life works rather than immediate application, laid the groundwork for modern applications in biology. Next, we discuss how evolution produced novel cell types with extraordinary properties and how leveraging these innovations can enable the creation of radical phenotypes beyond natural evolutionary constraints. Then, we explore the potential of artificial cells, constructed from the bottom up, as experimental platforms for studying genotype-phenotype relationships and testing evolutionary principles in real time. Finally, we argue that engineering radical phenotypes and emergent traits will require better investment in basic science, infrastructure, and graduate training to avoid bottlenecking innovation. History has shown that committing to basic science results in transformative solutions with far-reaching societal and economic benefits. By drawing inspiration from evolution and expanding synthetic biology beyond traditional model systems, we can push current boundaries to open new avenues for fundamental discovery and technological advances to stimulate the bioeconomy.

Variable thermal environments may have both detrimental and beneficial effects. For example, extreme temperatures may challenge homeostasis and inflict tissue damage but may also induce acclimation that improves stress resilience. Hormetic models provide a framework to understand dosage-dependent, contrasting beneficial and detrimental effects from physiological and ecological perspectives. We used a hormetic framework and associated quantitative models to investigate how a range of relatively cold, pre-exposure temperatures influence survival and fertility following cold shock at a more extreme cold temperature in the fruit fly Drosophila melanogaster . Cold pre-exposure can induce a protective rapid cold hardening (RCH) response, fail to stimulate a response, or cause direct cold injury. We found a plateau-shaped relationship between pre-exposure temperature and female survival resilience, where survival following a cold shock remains high across a range of temperatures, with sharp transitions at higher and lower temperatures. Bayesian fitting of a bi-logistic model highlights these transitions at temperature thresholds that govern processes mediating both acclimation and cold injury. In contrast to survival, female fertility resilience exhibited a muted response to pre-exposure temperature in the presence and absence of post-stress mating opportunities. Overall, a range of pre-exposure temperatures allowed low but successful reproduction following cold shock. High survival but low fertility resilience is consistent with a) differential impacts of cold on somatic and reproductive tissues and b) a growing body of literature suggesting that the thermal sensitivity of fertility may be more limiting than survival for population persistence in variable and changing climates. Summary statement A hormetic model shows how sharp temperature thresholds govern beneficial rapid cold hardening and detrimental cold injury that have well-defined effects on survival, but only weakly affect fertility. ### Competing Interest Statement The authors have declared no competing interest. National Science Foundation, 2045263

Mayaro virus (MAYV) is an alphavirus endemic in many parts of Central and South America transmitted to humans by Aedes aegypti. Currently, there is no vaccine or treatment of Mayaro infection, and therefore it is essential to control transmission by reducing populations of Ae. aegypti. Unfortunately, Ae. aegypti are extremely difficult to control with traditional integrated vector management (IVM) because of factors such as growing resistance to a dwindling list of registered insecticides and cryptic immature and adult habitats. The sterile insect technique (SIT) by irradiation is gaining traction as a novel supplemental tool to IVM. The SIT is being used operationally to release large numbers of sterilized colony-reared male mosquitoes in an intervention area to overwhelm females in the natural population, eventually causing population decline because of high frequencies of unfertilized eggs. However, little is known about the effect of irradiation on vector competence for mosquito-borne viruses such as MAYV in females that may be accidentally reared, irradiated, and released alongside males. In this investigation, we exposed female Ae. aegypti pupae to radiation and evaluated vector competence after inoculation with MAYV. Infection and dissemination rates of irradiated (10 and 40 Gy) Ae. aegypti were higher than those of non-irradiated cohorts at 7 and 14 days after infection. Although these results indicate a need to maintain effective sex sorting prior to irradiation and release of Ae. aegypti, our results are consistent with several previous observations that vectorial capacity and vector competence are likely lower in irradiated than in nonirradiated females.

The highly invasive Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), is currently expanding its geographic distribution into cooler temperate areas of the Northern Hemisphere. In marginal conditions, the invasion potential of medfly depends in part on innate tolerance to the novel environmental conditions. Physiological tolerances are potentially influenced by interactions among multiple factors, such as organism age or reproductive maturity, sex, and mating status. Furthermore, the relationships between the above factors and tolerances may differ among geographically distinct populations. Here, the effects of age and mating status on thermal tolerance of three geographically distinct medfly populations along a latitudinal gradient ranging from Greece (Crete & Volos) to Croatia (Dubrovnik) were examined. The upper and lower critical thermal limits (scored as loss of neuromuscular function during controlled cooling or heating) of adult males and females (a) at 1-, 6-, 15-, and 35 days old and of (b) both mated and virgin flies were assessed. Results showed that estimates of lower and upper thermal limits (CTmin and CTmax) were both population- and age-dependent. In most age classes tested, CTmin values were lower for the adults obtained from Crete and higher for those from Dubrovnik. CTmax values were lower for the females from Dubrovnik compared to the females from any other population on day one after emergence but not on days 6, 15 and 35. Differences among populations were observed across different age classes both for cold and heat tolerance but mostly in CTmin estimates. Mating status had a little effect on cold and heat tolerance. Complex patterns of thermal limit variation within and among populations suggest a suite of factors determine population-level mortality from thermal extremes under field conditions in medfly. These results contribute towards understanding the invasion dynamics of medfly and its range expansion to northern, more temperate regions of Europe.

Insects have major impacts on forest ecosystems, from herbivory and soil-nutrient cycling to killing trees at a large scale. Forest insects from temperate, tropical, and subtropical regions have evolved strategies to respond to seasonality; for example, by entering diapause, to mitigate adversity and to synchronize lifecycles with favorable periods. Here, we show that distinct functional groups of forest insects; that is, canopy dwellers, trunk-associated species, and soil/litter-inhabiting insects, express a variety of diapause strategies, but do not show systematic differences in diapause strategy depending on functional group. Due to the overall similarities in diapause strategies, we can better estimate the impacts of anthropogenic change on forest insect populations and, consequently, on key ecosystems.

The spread of arboviruses like yellow fever, dengue, chikungunya, and Zika, transmitted by the invasive mosquito Aedes aegypti has led to the development of many strategies to suppress mosquito populations. Given the rapid development of resistance to common chemical larvicides and adulticides in some Ae. aegypti populations, as well as the ever-shrinking chemical options for mosquito control, there is a pressing need for new tools and deployment of those innovative tools as a component of integrative mosquito management programs. Prior to the adoption of any mosquito population intervention, be it conventional or innovative, understanding the baseline population is essential to evaluate the efficacy of the control measure. The Lee County Mosquito Control District in Florida has collected a three-year-long period of baseline entomological surveillance data collection for Ae. aegypti on Captiva and Sanibel Islands as foundational information prior to implementation of a new integrative mosquito management approach. We identified 18 mosquito species and described their population dynamics during the rainy and dry seasons. The two islands had no significant differences in species richness, diversity, dominance, or evenness overall. Yet, there were clear differences between the high rain season and low rain season in the Shannon diversity index, Simpson dominance index, and Pielou species evenness index within each site. Our data suggest that any innovative intervention should begin before mid to late April when the mosquito population is at its lowest and certainly before populations build up to their summer peak between June and September. These data also show the spatial distribution of Ae. aegypti is dynamic in space and time, identifying hotspots of mosquito abundance to focus on for future interventions. Overall, our study emphasizes the importance of entomological data collection to understand the population dynamics of Ae. aegypti mosquitoes, including the impact of environmental factors such as temperature and precipitation.

During the winter, animals face limited food availability. Many animals enter dormancy to reduce their winter energy expenditure. Most insects spend the winter in diapause, a state of programmed dormancy. It is often assumed that diapausing insects need nutrient stores to fuel their many months of basal metabolism and must grow heavier than their non-diapause-programmed counterparts. However, the extent to which food limitation affects body weight during overwintering preparation as well as the likelihood and duration of diapause remains unclear. We limited the duration of the feeding period and thus the total quantity of food available to diapause-destined larvae of the pupal-diapausing flesh fly, Sarcophaga crassipalpis, to test how food limitation affects body weight in the context of diapause programming. We also tested the extent to which food deprivation and body weight affect the likelihood and duration of diapause. We hypothesized that diapause-destined larvae grow more quickly and pupariate at a heavier body weight than non-diapause larvae. We also hypothesized that body weight is more dramatically reduced by food limitations when a larva is programmed for diapause. Finally, we hypothesized that larvae with lighter body weight (i.e., food limited) are less likely to enter pupal diapause and also stay in diapause for a shorter duration than heavier, well-fed, individuals. Contrary to our hypotheses that diapausing insects are heavier than their non-diapausing counterparts, we found diapausing pupae weighed less than non-diapausing pupae, especially when larvae received limited food. We found light pupae did not abort their diapause program. In both diapausing and non-diapausing pupae, body weight was positively correlated with simulated winter survival. However, above a weight threshold, body weight no longer affected simulated winter survival in diapausing pupae. Contrary to our predictions and the general consensus in much of the diapause literature, we also found that lighter pupae stayed in diapause longer than heavier pupae. Overall, our results challenge the precept that body weight and diapause are positively associated. The relationship between body weight and diapause is complex and may be affected by the availability of food before and after winter, the availability of high-quality overwintering sites, and the life history of a particular insect. Our findings challenge the positive association between body weight and diapause (dormancy) phenotypes in the model flesh fly, Sarcophaga crassipalpis. Nutrient limitation alters the relationship between body weight and diapause phenotypes, suggesting that the relationships between body-weight and phenotypes need to be considered within ecological contexts.image

Genetic biocontrol approaches to insect pest management offer high species specificity and reduce insecticidal chemical pollution, but these approaches require the release of genetically modified organisms that must perform across a variety of environmental conditions. Because organisms often display phenotypic and gene expression variability when exposed to different environments, it is reasonable to expect that transgenic systems may be prone to environmentally mediated variation in expression and function. In this study, we examined the influence of two abiotic variables, temperature and nutrition, on the penetrance of an early embryonic Tet-off conditional lethality system in the vinegar fly, Drosophila melanogaster. We manipulated parental and offspring environments independently to determine the impact of life stage of exposure on transgene performance by estimating the probability of larval hatching and measuring transcript abundance of the transgenic system components. Our findings revealed that: 1) transgene performance has distinct norms of reaction to temperature and nutrition, 2) the effects of abiotic challenge are greatest when exposure occurs in the embryos expressing the transgene compared to parental exposure, and 3) stress exposures at the extreme limits of permissible conditions can dramatically decrease the penetrance of transgenic lethality. While variation in transcript abundance of the transgenic system was observed in some environments, these changes were not fully congruent with patterns of phenotypic penetrance, suggesting that the observed variation in lethality is likely driven by processes downstream of transcription.

Fat reserves, specifically the accumulation of triacylglycerols, are a major energy source and play a key role for life histories. Fat accumulation is a conserved metabolic pattern across most insects, yet in most parasitoid species adults do not gain fat mass, even when nutrients are readily available and provided ad libitum. This extraordinary physiological phenotype has evolved repeatedly in phylogenetically dispersed parasitoid species. This poses a conundrum because it could lead to significant constraints on energy allocation toward key adult functions such as survival and reproduction. Recent work on the underlying genetic and biochemical mechanisms has spurred a debate on fat accumulation versus fat production, because of incongruent interpretation of results obtained using different methodologies. This debate is in part due to semantics, highlighting the need for a synthetic perspective on fat accumulation that reconciles previous debates and provides new insights and terminology. In this paper, we propose updated, unambiguous terminology for future research in the field, including "fatty acid synthesis" and "lack of adult fat accumulation", and describe the distinct metabolic pathways involved in the complex process of lipogenesis. We then discuss the benefits and drawbacks of the main methods available to measure fatty acid synthesis and adult fat accumulation. Most importantly, gravimetric/colorimetric and isotope tracking methods give complementary information, provided that they are applied with appropriate controls and interpreted correctly. We also compiled a comprehensive list of fat accumulation studies performed during the last 25 years. We present avenues for future research that combine chemistry, ecology, and evolution into an integrative approach, which we think is needed to understand the dynamics of fat accumulation in parasitoids.

In North America, weather and host-plant abundance drive the population dynamics of the migratory pest Helicoverpa zea. The objectives of this study were to (i) estimate monthly abundance of H. zea moths in Bt cotton and peanut fields, (ii) document the effects of weather on H. zea trap catches, and (iii) determine larval hosts supporting H. zea populations from 2017 to 2019. Year-round trapping of H. zea moths was conducted in 16 commercial fields in two regions of the Florida Panhandle using delta traps. H. zea moth catches were associated with temperature, rainfall, and relative humidity. Larval hosts were determined by isotopic carbon analysis. Our results showed year-round H. zea flights in both regions across two years, with the highest and lowest moth catches occurring from July to September and November to March, respectively. There was no difference in catches between traps set on Bt cotton and peanut. In the Santa Rosa/Escambia counties, weather explained 59% of the variance in H. zea catches, with significant effects of temperature, relative humidity, and rainfall. In Jackson County, weather explained 38% of H. zea catches, with significant effects of temperature and relative humidity. Carbon isotopic data showed that feeding on C3 plants, including Bt cotton, occurred over most of the year, although feeding on C4 hosts, including Bt corn, occurred during the summer months. Hence overwintering and resident populations of H. zea in the Florida Panhandle may be continually exposed to Bt crops, increasing the risk for the evolution of resistance.

Climate change may alter phenology within populations with cascading consequences for community interactions and on-going evolutionary processes. Here, we measured the response to climate warming in two sympatric, recently diverged (~170 years) populations of Rhagoletis pomonella flies specialized on different host fruits (hawthorn and apple) and their parasitoid wasp communities. We tested whether warmer temperatures affect dormancy regulation and its consequences for synchrony across trophic levels and temporal isolation between divergent populations. Under warmer temperatures, both fly populations developed earlier. However, warming significantly increased the proportion of maladaptive pre-winter development in apple, but not hawthorn, flies. Parasitoid phenology was less affected, potentially generating ecological asynchrony. Observed shifts in fly phenology under warming may decrease temporal isolation, potentially limiting on-going divergence. Our findings of complex sensitivity of life-history timing to changing temperatures predict that coming decades may see multifaceted ecological and evolutionary changes in temporal specialist communities.