Ecological Entomology

To implement adequate nutritional landscapes to bumble bees and inform conservation efforts, we need a better understanding of how different pollen sources impact bumble reproductive performances and phenology. We assessed the impact of multiple native pollen on bees reproductive success by feeding 40 wild-caught Bombus impatiens and B. griseocollis queens with either commercially wildflower pollen mix commonly used to breed Bombus queens (control group) or 75% red maple pollen (Acer rubrum) and 25% pollen mix (RM group). In a second experiment, we monitored the reproductive success of 45 microcolonies of workers, harvested from B. impatiens colonies. Microcolonies were fed either: 1) pollen mix (N=15), 2) pollen mix and hawthorn (Crataegus canadensis, N=15), or 3) pollen mix and sumac (Rhus typhina, N=15). In experiment 1, RM significantly increased reproductive outputs for both species. RM-fed founding queens were twice more likely to produce workers, and produced them significantly earlier, though total number of workers did not vary between groups. However, RM-fed founding queens produced significantly more brood cells, males and gynes than control colonies, and produced them significantly earlier. In experiment 2, we found an interaction between initial colony diet and current microcolony diet. Overall, workers harvested from RM-fed colonies produced significantly more brood cells and males than workers harvested from control colonies. However, when fed hawthorn pollen during experiment 2, workers from control colonies performed as well as workers from RM-fed colonies. This suggests carry-over effects from early pollen sources provided to colonies on workers, which may be circumstantially offset by pollen sources with specific nutrient profile. Using detailed nutritional information of each pollen (including data on 32 nutrients), we provide nutritional requirements of pollen to support bumble bee reproduction, especially regarding vitamin B3 and vitamin C contents. This study will help inform land managers and captive breeding programs on the ideal pollen sources for bumble bees to ensure reproductively successful colonies.

Riparian environments are characterised by a high diversity of arthropod species, linking the aquatic with the terrestrial ecosystem. One of the dominant arthropod predators in this ecotone, carabid beetles are of particular interest as they may also act as facultative scavengers, feeding on carrion deposited along the shoreline. To test whether carabids feed on carrion, we examined the feeding preferences of the abundant riparian carabid Bembidion elongatum in a laboratory feeding trial, offering freshly killed and 24 hours post mortem Drosophila melanogaster. We subsequently assessed the detection probability of ribosomal prey RNA and DNA in predator gut contents using Drosophila-specific RT-PCR and PCR assays, and quantified nucleotide abundance by quantitative real-time PCR at 0, 3, 6, and 12 hours post-feeding. In the feeding experiment, B. elongatum showed a significant preference for fresh over carrion prey. Following consumption, the quantities of prey DNA and RNA in the predators gut declined over a 12-hour post-feeding period. However, no differences were detected in prey DNA or RNA quantities between individuals fed fresh prey and those fed carrion. Only immediately after consumption was the DNA:RNA ratio significantly lower in individuals fed fresh prey compared to those fed carrion while this difference was not observed at later time points. Overall, our results indicate that ingested ribosomal prey RNA in predators is present in high quantities, and that the DNA:RNA ratio is not a suitable indicator for distinguishing between consumption of carrion and fresh prey.

Understanding how closely related species coexist in highly seasonal and unpredictable environments is central to studies of ecological differentiation and niche partitioning. We investigated the feeding ecology of sympatric populations of Microcebus murinus and M. griseorufus within a contact zone in Andohahela National Park, southeastern Madagascar, across dry and wet seasons. Using a combination of direct behavioral observations (1,611 feeding records), fecal sample analyses (n = 56), and vegetation phenology surveys, we quantified dietary composition, seasonal shifts in resource use, and habitat-related variation. Seasonal changes in diet were pronounced, with dry-season feeding dominated by exudates and wet-season diets incorporating greater proportions of fruit and flowers, closely tracking phenological patterns at both sites. Diets of both species were dominated by plant resources, but consistent interspecific differences in dietary strategy were evident. Although both species consumed comparable proportions of insect prey, M. murinus showed pronounced wet-season increases in the use of high-sugar, carbohydrate-rich floral resources (15.9%) and hemipteran-associated honeydew (30.6%). In contrast, M. griseorufus relied more consistently on predictable exudates throughout the year. Fecal analyses supported observational data but revealed differences in the detectability of dietary components, with increased representation of invertebrates in the wet season and seeds in the dry season. These results indicate substantial dietary overlap but consistent differences in resource use, suggesting that coexistence is facilitated by fine-scale trophic differentiation within a broadly shared omnivorous niche. Such subtle but persistent differences in feeding strategy likely reduce competitive overlap and enable continued sympatry in a climatically variable and resource-limited system.

Many insect populations are experiencing thermal stress as a result of global change, making it imperative to investigate how their relationship with other organisms will be impacted by heat disturbances. Microbial symbionts, such as bacteria, have the potential to enhance or inhibit an insect's thermal tolerance. Solitary bee larvae host bacteria within their food stores ("pollen provisions"), which have been shown to benefit survival and development; however, it is unclear how heatwaves brought about by climate change will impact their relationships with these bacterial partners. In this study, we subjected blue orchard bee (Osmia lignaria) eggs and larvae to a 4-day heatwave (35 {degrees}C daytime:22 {degrees}C nighttime) or kept them at control temperatures (25 {degrees}C daytime:15 {degrees}C nighttime), then returned all bees to control temperatures for a 5-day recovery period. We assessed bacterial communities within pollen provisions and larval development stage pre-heatwave (Day 0), immediately post-heatwave (Day 4), and following the recovery period (Day 9). Bacterial community composition, diversity, and abundance were resilient to heat stress, but larval bees developed faster when subjected to a heatwave. This finding refutes the hypothesis that bacteria within pollen provisions modulate blue orchard bee responses to heat, suggesting instead that developmental effects could be more largely shaped by bee physiology or interactions with microorganisms other than bacteria.

Wildflower plantings are an important conservation measure for supporting wild bee diversity. They aim to enhance floral resources for nutrition, but do not explicitly consider that bees also require suitable nesting and overwintering resources. Wildflower plantings may provide nesting habitat for ground-nesting bees, but the effects of soil management, e.g. ploughing, on ground-nesting bees are hardly known. To study how ploughing and age of wildflower plantings affect ground-nesting bees, we sampled bees on ploughed and unploughed wildflower plantings aged 0-4 years. We used emergence traps to sample bees directly after emergence from the ground, allowing inference on nest numbers. We found that ploughing and wildflower planting age negatively affected overwintering bee nest numbers. Nesting peaked in the year of wildflower planting establishment and declined thereafter, indicating lower habitat suitability at later successional stages. Annual ploughing caused the greatest reduction in nest numbers (-72 %), providing evidence for an ecological trap.

Understanding the spatial distribution of species and their patterns of endemism is necessary for establishing effective conservation priorities. Despite the vital pollination services bees provide, Californias native bee distribution patterns remain largely unexplored relative to plants and butterflies. We analyzed bee species richness and endemism across California and their concordance with plant distributions. Richness was high across areas of the California Floristic Province, including the Sierra Nevada, San Francisco Bay Area and Central Coast, South Coast Ranges, and the Transverse and Peninsular ranges. Bee endemism was more localized, concentrated in the San Joaquin Valley, eastern Sierra Nevada and adjacent Great Basin, Sierra Nevada foothills, and California deserts. Because richness and endemism appear to operate at different spatial scales and likely respond to different environmental drivers, effective conservation strategies must address both. Additionally, conservation plans that incorporate both plant and bee diversity are needed to achieve more comprehensive biodiversity protection.

The expansion of global trade has increased the risk of unintentional introductions of woodboring insects outside their native ranges, which can cause considerable ecological and economic damage. Monitoring efforts for native and non-native species of woodborers rely heavily on semiochemical-baited traps. With the goal of improving detection efforts of longhorned beetles (Coleoptera: Cerambycidae) and their associates, we conducted field bioassays in loblolly pine dominated and bottomland hardwood forest ecosystems, which are representative of subtropical Louisiana. We tested the impact of attractant composition, trap design, and the application of a friction reducing compound on the capture of longhorned beetles. We compared the Synergy Multi-Trap System with and without Fluon, and the Synergy Funnel Trap II with Fluon, baited with multicomponent cerambycid lures and ethanol or ethanol alone. In total, we collected 5,303 beetles comprising 44 species. Traps baited with cerambycid lures and ethanol captured ~11X more individuals than the traps baited with ethanol. We identified novel attractants for 6 species of longhorned beetles. When Fluon was applied, both the Synergy Multi-Trap System and Funnel Trap II were equally effective at capturing cerambycids. Application of Fluon significantly increased the trap capture of the Synergy Multi-Trap System by ~4X compared to the same trap type without Fluon. Our results contribute to our understanding of the factors influencing trap captures and the chemical ecology of woodboring insects thereby helping the development of early detection and rapid response monitoring programs in subtropical regions of the United States and elsewhere.

Population dynamics and their component vital rates may be driven by weather, climate teleconnections between sea and air (e.g. ENSO), or biotic interactions. These drivers operate directly or indirectly and on different temporal scales. We used a Bayesian structural equation model to characterize effects among weather, climate, and incidental intraguild predation (IGP) on the butterfly Euphydryas gillettii's vital rate of pre-diapause survival, using an 18 year dataset. IGP was a major determinant of pre-diapause survival, along with direct and indirect effects of weather and spring climate teleconnections. The direction of climate effects was reversed when mediated through IGP. Our analysis illustrates the need for sequential hypotheses to capture the cascading effects of abiotic factors via biotic interactions. Using sequential hypotheses addresses the debate on weather -- climate teleconnection roles by disentangling their contributions from one another. Finally, vital rates must be decomposed to component rates in order to detect their drivers.

The ecological and economic threat posed by invasive pests demands proactive mitigation. Species distribution models (SDMs) are widely used in efforts to predict where invasive species might spread after introduction, though such models face several limitations. Among these is the unrealistic assumption of niche uniformity throughout a species' range. This has led to interest in developing SDMs that explicitly account for local adaptation, though few methods have achieved this in a way that confidently separates local adaptation from population structure. Here we develop and implement a sequential SDM approach that incorporates experimentally verified associations between genotype, phenotype, and environment to forecast establishment risk in a major agricultural pest. We leverage genomic data from 738 individuals to characterize the geographic distribution of alleles at a major-effect locus for cold tolerance (tret1) in Helicoverpa armigera, an invasive crop pest of major economic concern in North America. We demonstrate that a recently detected North American population carries a cold-adapted tret1 allele, which has likely contributed to its persistence. We quantify the contribution of cold-adapted tret1 to the potential invasive range of H. armigera in North America under current and future climate scenarios. We find that cold-adapted tret1 may dramatically expand the potential range of H. armigera, and that potential future range expansion is likely to be driven primarily by cold-adapted individuals. Our results highlight the importance of accounting for intraspecific variation in invasive species risk assessments and management strategies.

Land-use change, such as the transformation of woody ecosystems into open pastures, acts as a strong ecological filter, favouring some species while excluding others according to differences in ecological niche breadth. Understanding how differences in niche breadth influence species responses under anthropogenic filters is crucial to anticipate their persistence or displacement. In this study, we quantified realized niche breadth in two sympatric ecosystem engineers, the Neotropical leaf-cutter ants Atta cephalotes and Atta laevigata, to test whether breadth differences are consistent with specialist and generalist ecological strategies. We characterized realized niche breadth across fine-scale environmental gradients by integrating hemispherical photography, microclimatic data, mound architecture, and edaphic profiles from 114 colonies across a regional transect in the Colombian Andes, alongside macroclimatic data from Copernicus. Principal Component Analysis (PCA) and PERMANOVA identified canopy openness and bushes- and tree-type vegetation density as the principal axes of interspecific niche partitioning. The observed differences in realized niche breadth were consistent with specialist and generalist ecological strategies. A. laevigata was predominantly associated with open-canopy areas, warmer micro- and macroclimatic conditions, and narrower edaphic dispersion. In contrast, A. cephalotes occupied a wider range of microhabitat conditions. This broader realized niche breadth is compatible with previous reports of A. cephalotes occurring in urban areas. Together, these findings suggest that niche breadth may influence how Neotropical leaf-cutter ants respond to habitat transformation, helping to understand the ecological consequences of land-use change.

Protected areas are vital to the recovery of endangered species. Of the 24 remaining endemic passerines in Hawaii, 16 species are endangered or critically endangered. Yet protected areas in the archipelago are changing as a result of climate change and biological invasions. For example, the non-native southern house mosquito (Culex quinquefasciatus), the primary vector of avian malaria (Plasmodium relictum), has been encroaching upward as higher elevations warm. How the ranges of endemic birds have also shifted their range upward in the Ka[u] Rainforest, the largest native forest on the Island of Hawaii, is not well understood. We used hierarchical distance sampling to characterize how population density has changed from 2002 to 2024 for eight endemic bird species in the Ka[u] Rainforest. For five species, the most parsimonious model included an interaction between year and a quadratic elevation effect. Species that were most common at lower elevations in 2002, e.g., Apapane (Himatione sanguinea), tended to be most common at mid-elevations by 2024. Species that were already most common at higher elevations, such as Iiwi (Drepanis coccinea) and Hawaii [A]kepa (Loxops coccineus), tended to decline in density. For example, Iiwi density at 1,530 m declined from 3.31 birds per ha (95% confidence interval [CI]: 2.67-4.11) to 1.34 birds per ha (95% CI: 1.05-1.71), and Hawaii [A]kepa were completely extirpated from elevations below 1,530 m by 2024. Our results demonstrate how the elevational ranges of endemic species have shifted in response to climate change, biological invasions, and habitat degradation. Translating these results into conservation measures may require a more thorough investigation of the causal mechanisms of these range shifts with finer scale habitat data, under the same hierarchical modeling framework.

Hibernation is a critical phase in the annual cycle of boreal bats, especially near the northern limit of their range where winter conditions are severe and suitable hibernacula are limited. In Finland, anthropogenic structures, such as abandoned bunkers, commonly serve as hibernacula for bats. In this research, we investigate how the size characteristics of bunkers affect species composition and abundance of hibernating bats. The length of the bunker was the best predictor of the presence and abundance of bats. The longest sites ([~]25-26 m) had the highest probability to host all studied species, or group of species of bats, specifically Eptesicus nilssonii, Plecotus auritus, and Myotis bats. Eptesicus nilssonii appeared to be a generalist species, utilizing a variety of bunkers regardless of their length or temperature regime. Still, its number per bunker was relatively low. The highest numbers of Myotis bats were observed in the longest bunkers with stable temperature. The number of P. auritus was positively related to the bunker length; however, it was rare in the studied hibernacula. Different species showed different patterns in hibernacula space usage. Eptesicus nilssonii hibernated in any part of the bunker, while Myotis bats used the most remote from the entrances areas in small sites, and occupied the whole area of the longest bunkers.

Social wasps (family: Vespidae) are increasingly concerning invaders and have been subject to increased detections and a growing number of invasive populations in the last few decades. As established invasive populations are challenging to eradicate, preventing introductions and prioritizing early interventions are the most cost-effective management solutions to mitigate these effects. A current challenge to this approach is that species distribution data is limited for many social wasp species, hindering our ability to accurately predict novel habitats with high suitability. To address this gap, we used MAXENT to create species distribution models (SDM) for 299 species of social vespid. We identified existing invasive populations of social wasps and incorporated their current invasive ranges to improve the transferability of our models in predicting habitat suitability in new environments. Current range sizes and habitat suitability varied widely among species and genera. We identified new species of high invasive concern, particularly in the genus Vespa. We also identified previously unrecognized regions that may be at high risk of future invasion primarily in Central Africa and the Indo-Australian Archipelago. Combining current and suitable ranges, we calculated an "Invasion Risk Score" to compare the relative likelihood of each species establishing a new invasive population based upon habitat suitability. To assess invasion risk in the future, we projected habitat suitability under four Shared Socioeconomic Pathway (SSP) climate change scenarios. Under all scenarios, species faced significant changes in habitat suitability for current native ranges. Habitat suitability generally shrank and shifted towards the poles, leaving equatorial species at highest risk of habitat loss. Notably, Vespa was the only genus whose suitable habitat expanded under these climate scenarios. Our framework demonstrates how multi-species SDMs can be applied to risk management of invasive populations.

Heritable bacterial symbionts are pervasive in terrestrial arthropods, often imposing reproductive manipulations to promote their own spread within host populations. Co-infections are common, potentially allowing symbiont co-infectors to hitchhike through a host population. However, adverse thermal conditions can disrupt these communities, particularly when co-infectors vary in their thermal sensitivity. We used a multi-generation experiment to test whether warm (29 {degrees}C) conditions disrupted spread of heritable symbionts through uninfected populations of the spider, Mermessus fradeorum. We tested two common infection combinations: a single infection with a cytoplasmic incompatibility (CI) inducing Rickettsiella or a feminizing co-infection that included a feminizing Wolbachia, the same Rickettsiella, and up to three apparent hitchhikers (two additional Wolbachia strains and Tisiphia). We initiated replicate populations with 1/3 of one infection type and 2/3 uninfected spiders, evaluating population infection rate over 5 spider generations under different temperature regimes. Under cool (21{degrees}C) conditions, Wolbachia feminization drove co-infection to 88% and Rickettsiella CI drove single infection to 83% of host populations. Vertical transmission for all symbionts was high (97-99%) and hitchhiking symbionts also spread effectively. Under warm conditions, feminization and CI efficacy were reduced, and symbionts suffered variably reduced vertical transmission. Warm conditions ultimately destroyed the co-infecting symbiont consortium and impeded symbiont spread. On its own, though, Rickettsiella was still able to increase, despite reduced strength of CI. We hypothesize that contrasting tensions between feminizing spread of the symbiont consortium versus environmentally driven loss of function and transmission may explain observed patterns of mixed infections in field populations of this spider.

Predicting species distributions under climate change typically relies on thermal limits for survival. However, recent evidence suggests that sublethal temperatures that reduce fertility can constrain distributions more strongly than temperatures causing mortality alone. Despite the importance of mosquitoes as vectors of human disease, thermal fertility limits remain largely uncharacterized in these insects. Here, we show that fertility in Aedes aegypti is highly sensitive to thermal stress and exhibits distinct sex-specific responses. Adult males were more vulnerable to both heat and cold exposure than females. At 38{degrees}C, survival remained high in both sexes despite significant reproductive impairment, indicating that sterility can occur independently of mortality. Similarly, exposure to 2{degrees}C induced male sterility whereas females maintained reproductive function. Morphological analyses of reproductive organs showed temperature-associated damage consistent with the observed fertility loss. Incorporating these fertility thresholds into ecological niche models generated different predictions of climatic suitability compared with models based solely on lethal temperature limits. Our findings demonstrate that sublethal effects on reproduction can substantially influence estimates of mosquito climatic suitability and highlight the importance of incorporating fertility based thermal limits into projections of vector distributions and disease risk under future climate change.

Animals often attempt to resolve the trade-off between foraging and predation by regulating their activity timings and patterns. Driven by individual energetic requirements and the local environment, onset of activity in a species varies across space and time. These differences are particularly enhanced on a latitudinal gradient, influenced by varying lengths of daylight, temperature and seasonality. For a nocturnal mammal like the bat, such patterns translate to shorter activity windows at higher latitudes, both on a nightly and a seasonal basis. This constructs the timing of activity onset as a crucial variable for a successful night of foraging. While roost exit timings and influence of local conditions on bat activity have been extensively studied across species inhabiting a variety of locations around the world, our understanding of bat emergence behaviour on a global scale remains limited. To investigate emergence patterns in bats across a latitudinal gradient, we conducted a meta-analysis spanning 91 species from across 13 Chiropteran families. We evaluated how timing and luminosity (i.e., sun altitude) at exit varied across latitudes and species. Results revealed that bats across the globe tended to emerge under conserved thresholds of brightness, a pattern that was facilitated by a temporal delay in roost exits at higher latitudes. By maintaining emergence within the confines of nautical twilight (sun altitude between 0{degrees} to -12{degrees}), bats across the phylogeny thus seemed to balance the dilemma of predation versus foraging. This pattern was mirrored across the two broad dietary types (frugivores and insectivores) as well as different social structures (maternal v. non-maternal colonies). Furthermore, our models indicated that while local conditions dictate final exit decisions, shared ancestry was potentially influencing the extent of plasticity seen in emergence patterns. Taken together, these results highlight how broad-scale latitudinal gradients of light can influence the foraging strategies in a globally distributed nocturnal mammal.

Assessing breeding phenology using singing intensity can potentially provide insights into a species' responses to climate or habitat change. Although male passerines are known to sing elaborate songs during the breeding season, females of several tropical passerines also sing. The Nilgiri Flycatcher (Eumyias albicaudatus) is an endemic dimorphic songbird found in the southern Western Ghats of India. Despite its limited distribution and its ecological significance as an indicator species of cloud forests, very little is known about its vocal behaviour and breeding ecology. To examine sex-specific differences in vocalisation patterns, we analysed 294 male and 102 female focal songs from two pairs of birds. We then used data from a year-long automated recorder placed near a breeding pair to study the species' singing phenology using the BirdNET Analyser. The two sexes broadly produced similar songs, although males' songs included more notes at a faster pace. The sex-specific detectors failed due to this high similarity among songs of the two sexes, but we were able to automate species-level detection (F1-score = 0.795 at a confidence threshold of 0.1). We found strong seasonality in vocal activity, with peaks in singing during the breeding season, and identified a diurnal peak at dawn. More data are required to parameterise differences in annual vocal activity between the sexes. These detection patterns provide a framework for a landscape-wide examination of Nilgiri Flycatcher phenology and occurrence. This study suggests that automatic detectors can be a reliable tool for assessing singing phenology in birds and provide insights into breeding periods.

Wildlife monitoring in arid environments is complicated by substantial temporal and spatial variation in animal activity driven by unpredictable environmental conditions. For highly mobile species, movements across the landscape may further influence activity recorded at fixed monitoring locations, raising questions about whether short-term surveys adequately represent patterns of site use. We examined temporal variation in acoustic activity of the threatened Pilbara Leaf-nosed Bat (Rhinonicteris aurantia Pilbara form), a highly mobile, cave-roosting insectivorous bat inhabiting the arid Pilbara region of north-western Australia. Acoustic activity was monitored continuously at two permanent diurnal roosts for up to two years, comprising 1601 detector-nights across three detector locations. We tested relationships between nightly activity and environmental conditions, detector location and anthropogenic disturbance. Activity varied substantially through time and among detector locations. Moon illumination and cumulative rainfall were strongly associated with activity, but their interaction differed between roosts: activity was highest under low moon illumination and low cumulative rainfall at Chateau Cave, whereas activity at Daltons was highest under low moon illumination and higher cumulative rainfall. Activity also differed markedly between detector positions within Chateau Cave, and humidity, temperature and artificial light at night had additional location-specific effects. Other measures of anthropogenic disturbance, including cave entry, blasting and mining activity, had comparatively weak support. The contrasting environmental relationships among locations demonstrate that acoustic activity at permanent roosts is strongly context dependent. For a species capable of extensive movements among roosts, temporal variation in activity may reflect behavioural responses and redistribution of individuals across the landscape rather than rapid demographic change. We suggest that facultative nomadism may provide a useful hypothesis for understanding temporal variation in roost use by Pilbara Leaf-nosed Bat. More broadly, our results demonstrate that short-duration acoustic surveys of highly mobile species in arid environments may provide an incomplete representation of longer-term site use and should incorporate temporal replication and environmental context when used for conservation and impact assessment.

Animals reproduce across a spectrum from oviparity to viviparity. Internal gestation and live birth have inherent advantages despite their costs. However, comparative analyses of key drivers, such as resilience to dehydration, lack taxonomic breadth. In this study, we assessed whether live birth improves the ability to proliferate in environments with limited access to water. To do so, we used the viviparous cockroach, Diploptera punctata, as a model of viviparity to assess dehydration-induced damage during extended periods of water deprivation. During pregnancy, maternal survival did not decline during dry periods compared with non-pregnant individuals, suggesting that pregnancy has minimal impact on resistance to dehydration stress. When mothers are deprived of water for 2 weeks, they show an increase in osmolality after one week, while their embryos show no change in osmolality until after two weeks, at which point abortions occur. Periods of dehydration increased the duration of pregnancy, likely due to a slower production of in utero milk, but there was no change in the size of progeny. After birth, viviparous D. punctata newborns showed increased survival under dry conditions compared to two other ovoviviparous species, which are much smaller and dehydrate more quickly. The combined impact of increased dehydration resistance during pregnancy and during the first independent phase indicates a benefit of prolonged viviparity in Diploptera punctata. This study provides evidence that viviparity in animal systems could enable progeny survival during drought.

Heliococcus summervillei is an emerging invasive mealybug that causes severe dieback in grasses in pastures and turfgrass landscapes. It is widespread in Australia and has recently been detected across the Caribbean, Mexico, and the United States. Accurate identification of mealybugs is challenging due to cryptic morphology, overlapping diagnostic characters, and limited taxonomic expertise and literature, which makes molecular tools essential for regulatory diagnostics and management. We developed the first Cytochrome Oxidase I (COI) barcode for H. summervillei and used it to examine mitochondrial variation across available populations. COI sequences reveal approximately a 10.2% mitochondrial split between the Type A and Type B variants. Phylogenetic, haplotype network, and genetic distance analyses show that all invasive range populations share one haplotype associated with a recent invasion in the United States, Australia, Pakistan, and the Caribbean, whereas the Barbados lineage contains two closely related haplotypes that represent a historically stable mitochondrial variant. Together, these results establish the first COI reference library for H. summervillei, clarify mitochondrial lineage structure, and provide a practical barcode tool that enables rapid identification of invasive populations and supports timely regulatory and pest management responses. Recognizing mitochondrial variants also establishes a framework for resolving lineage-specific biological and management traits and strengthens reconstruction of introduction pathways central to regulatory decision-making and limiting further spread.