Insect Science

Spodoptera frugiperda is one of the most important agronomical pests due to its migratory capacity and broad host range. Since it is resistant to several insecticides, novel control strategies are being explored to control it. In this way, Spodoptera frugiperda Multiple Nucleopolyhedrovirus, a natural pathogen, has been proposed for its biocontrol. In this work, we performed a small RNA-seq on uninfected larvae and larvae infected with SfMNPV to identify expressed miRNA, characterize them, and identify differentially expressed (DE) miRNA in the infected condition. We identified several known and putative novel miRNAs, some of which are encoded in multiple copies and may be expressed within miRNA clusters. We also found 13 DE miRNA, most of them previously reported, two of them are putative novel miRNAs identified in this work. We predicted miRNA targets and found that their putative biological role could be related with processes relevant to the infection such as proliferative and apoptotic pathways, cell cycle regulation, autophagy, DNA damage response (DDR), vesicle transport, cytoskeleton remodelling, JAK/STAT and Toll signaling pathway, and immune response activation, among others. Moreover, we observed that several of the putative targets were hub genes in a predicted protein - protein interaction network. Finally, we found DE miRNA putatively associated with the regulation of viral gene expression, suggesting they might have a role in modulating the infection. Our results contribute to better understanding the miRNA landscape in S. frugiperda, and their putative role upon SfMNPV infection.

Behavioral thermoregulation allows animals to reduce the physiological costs associated with variation in the thermal environment, yet little is known about how chimpanzees respond behaviorally to the cool and humid conditions of tropical montane forests. We examined how microclimatic conditions influenced resting posture, sun/shade use, substrate use, vertical position within the canopy, and day nest use in eastern chimpanzees (Pan troglodytes schweinfurthii) in Nyungwe National Park, Rwanda. From March 2024 to February 2025, we collected 5,661 individual behavioral records during 486 contact hours and paired behavioral observations with measurements of ambient temperature, water vapor pressure, wind speed, and rainfall. Bayesian mixed models showed that temperature was the most consistent climatic correlate of resting behavior. Under cooler conditions, chimpanzees were more likely to adopt heat conserving postures, particularly curled and huddled postures, and to use day nests. As temperature increased, chimpanzees increased their use of shade, shifted from arboreal to terrestrial resting, and used lower canopy strata more frequently. Higher wind speed was also associated with reduced use of the upper canopy, whereas rainfall, water vapor pressure, and fruit availability generally showed weaker or behavior-specific associations. These findings indicate that chimpanzees in a cool montane forest flexibly adjust posture, solar exposure, substrate use, vertical position, and day nest use in relation to local thermal conditions. Resting behaviour may therefore provide an important context for behavioral thermoregulation, extending our understanding of chimpanzee thermal ecology beyond the warmer lowland and savanna habitats that have received most previous attention.

1. Most species are neither discovered nor named, let alone included in analyses that require biological information such as trait measurements, images, ecological information and genome-scale data. Specimen-level DNA barcoding can help discover many of these species rapidly, but everything beyond discovery requires vouchers organized into putative species. Yet, existing barcoding workflows lack efficient techniques for voucher recovery, creating a post-barcoding bottleneck that limits the ability of converting barcoded specimens into biological knowledge. 2. Here we present a low-cost, open-source workflow consisting of two stages. The first safeguards barcoded specimens by separating them from DNA extracts and transferring them from microplates into ethanol-filled glass vials. The second converts the resulting voucher collection into a searchable physical resource by linking barcode-derived molecular Operational Taxonomic Unit (mOTU) assignments to vial positions and enabling specimens to be sorted into putative species either manually or automatically using a newly developed open-access robot (SORTER). 3. We evaluated the workflow using 2,024 insect specimens distributed across 21 96-well plates. For the first stage, DNA separation and specimen transfer required approximately 15 minutes per plate. For the second stage, MOTUmapper generated retrieval coordinates in a few seconds, after which the 2,024 vouchers belonging to the 452 putative species could be recovered manually in 5 days or with SORTER in 5 hours. Throughout both stages, specimen identities remained linked to barcode sequences, metadata and storage positions. 4. Vouchers are the Rosetta stones of biology because they connect different kinds of data to the same specimens. By safeguarding these vouchers and making them searchable, the workflow converts barcode projects from one-time molecular surveys into reusable resources for ecological and evolutionary research.

BACKGROUND: Varroa destructor is the major ectoparasite of honey bees and a vector of viral pathogens. Because pathogen transmission and exposure to contact-active acaricides depend on mite host contacts, understanding the factors governing host residence time is important for both disease epidemiology and pest management. We quantified host residence time under varying bee densities and host-type compositions. RESULTS: Mean residence time was 9.48 h across 312 host-residence events. Mites remained on individual hosts for only 2.36 h on Day 1 but approximately 11-14 h from Day 2 onward. A generalized linear mixed model showed a strong positive effect of day on residence time ({beta} = 0.341, SE = 0.044, P < 0.001), corresponding to an approximately 41% increase in residence time per day. Excluding Day 1 eliminated this effect (P = 0.16), indicating that the temporal pattern was driven primarily by the initial exposure period. Reconstructing Day 1 observations to an 8-hour schedule confirmed that this pattern was not an artifact of observation frequency. Neither host type nor bee density affected residence time, and mite occupancy of nurse bees matched host availability. CONCLUSION: Host residence time was governed primarily by initial exposure rather than host identity or moderate crowding. The results identify a previously undescribed exploratory phase immediately after mites enter a novel adult-bee population. Because shorter residence times imply more frequent host switching, these findings improve our understanding of pathogen transmission dynamics and may help explain variation in the performance of contact-based Varroa control strategies.

This study establishes a comprehensive genomic, transcriptomic, and proteomic foundation for the wild cochineal bug, Dactylopius opuntiae, a scale insect and agricultural pest of prickly pear cacti notable for its dense white waxy covering. Although fatty acyl reductases (FARs) are central to insect epicuticular wax biosynthesis, the genes underlying wax production in cochineal insects are largely unexplored due to a lack of genomic resources. We report a 359-Mb de novo genome assembly in which we identify 26 FAR genes. Through phylogenetic reconstruction of Coccoidea FAR enzymes we reveal lineage-specific expansions of tandemly arranged D. opuntiae FAR genes, and discuss these enzymes roles in the development of this insects epicuticular waxy coating.

Insects of the subfamily Triatominae, commonly known as kissing bugs, are obligate blood-feeding vectors of Trypanosoma cruzi, the causative agent of Chagas disease. Rhodnius prolixus is among the most epidemiologically important vectors in Latin America, whereas Triatoma rubida frequently invades homes and is a potential vector in the southern United States and northern Mexico. Triatomines likely evolved from predatory reduviid assassin bugs through a transition from feeding on arthropods associated with vertebrate hosts to feeding directly on vertebrate blood. To investigate the genomic basis of this ecological and dietary shift, we generated highly contiguous, near chromosome-level genome assemblies and structural gene annotations for R. prolixus and T. rubida. The new R. prolixus assembly improves scaffold N50 more than 40-fold over the current reference genome, from 1.1 to 43.9 Mb, while reducing assembly gaps by several orders of magnitude. Both assemblies exceed 97% BUSCO completeness. Comparative analyses with representative hemipteran genomes revealed expansions of gene families associated with chemosensation and metabolism, including detoxification, protein degradation, and digestion, together with signatures of positive selection in genes involved in digestive and sensory functions. These assemblies represent the most contiguous and complete genomic resources available for Triatominae and provide a robust foundation for investigating vector biology, host adaptation, and the evolutionary origins of blood feeding within Reduviidae. Interpretive summaryKissing bugs are insects that are known for feeding on blood. They can spread a disease called Chagas disease because they transmit a parasite called Trypanosoma cruzi. To understand how kissing bugs evolved and which genes facilitate blood feeding of vertebrates, a collaboration between scientists at USDA-ARS, University of Georgia, and University of South Bohemia sequenced the genome of two kissing bugs: Rhodnius prolixus and Triatoma rubida. By comparing the genes with those of other insects in the order Hemiptera, scientists discovered that kissing bugs have more genes involved with detecting environmental chemical stimuli and metabolism as well as positive selection for genes involved with digestion and sensory-related proteins. These genome assemblies will help scientists learn more about how these insects evolved, and this research is important for understanding insect feeding biology which can be used to develop methods to control the kissing bugs and the spread of Chagas disease.

Honey bees can swim on the water surface toward dark regions, a behavior known as scototaxis that may facilitate escape from water. Although this behavior has been reported in both honey bees and solitary bees, variation among honey bee species remains poorly understood. We compared scototaxis during swimming in four honey bee species representing two nesting types: open-nesting (Apis florea and A. dorsata) and cavity-nesting (A. cerana and A. mellifera). Individual bees were released into a water-filled arena containing a dark sector, and their landing angles were recorded. All species exhibited significant orientation toward the dark sector. However, open-nesting species showed significantly stronger orientation than cavity-nesting species. No significant differences were detected between replicate colonies within species or between species within the same nesting type, whereas differences between nesting types were highly significant. Hierarchical clustering based on orientation strength placed Osmia, a solitary cavity-nesting bee from our previous study, in the same behavioral cluster as the two open-nesting Apis species rather than the cavity-nesting honey bees. We also measured swimming duration, distance, and velocity, but found no consistent differences between nesting types. These results demonstrate substantial interspecific variation in swimming scototaxis. The behavioral clustering is consistent with the hypothesis that strong scototaxis represents an ancestral trait that has been reduced in the derived A. cerana/A. mellifera lineage.

Japanese encephalitis virus (JEV) is an emerging mosquito-borne flavivirus with potential for geographic expansion, yet the risk of establishment in North America remains poorly characterized. We assessed vector competence of three North American Culex species (Cx. pipiens, Cx. quinquefasciatus, and Cx. tarsalis) for the JEV Nakayama strain, isolated from human brain in 1934 in Japan, across five constant temperatures (15, 20, 25, 30, and 33{degrees}C) at 4, 7, and 14 days post-feeding, quantifying infection, dissemination, and transmission rates. Vector competence was low but non-zero across all species. Cx. pipiens showed higher infection rates than the other species, whereas Cx. quinquefasciatus and Cx. tarsalis were minimally susceptible under these experimental conditions. Temperature had limited effects on infection and no detectable effects on dissemination or transmission. These findings suggest limited transmission potential of JEV Nakayama in North America, with Cx. pipiens as a relatively permissive vector.

BackgroundResistance of the arbovirus vector Aedes albopictus to pyrethroid insecticides is an emerging threat to vector control programs in Europe. Three knock-down resistance (Kdr) mutations affecting the voltage-gated sodium channels targeted by pyrethroids are known to confer resistance: V1016G, I1532T and F1534C. As these Kdr mutations are actively circulating in Europe, their monitoring is crucial for resistance surveillance programs. However, current Kdr genotyping methods are labor-intensive and costly, limiting their applicability for large-scale high-throughput surveillance. MethodologyNovel digital droplet PCR (ddPCR) TaqMan assays were developed, allowing the quantification of these Kdr mutations from pooled mosquito samples from field populations. The specificity of these assays was validated by using mosquitoes of known genotypes together with synthetic DNA constructs carrying haplotype combinations previously unseen in the field. The assays accuracy was assessed by comparing Kdr frequencies measured from pooled mosquitoes to those derived from individual genotypes. These assays were then used in a pilot surveillance study in mainland France, integrating deltamethrin bioassays, pooled Kdr mutation tracking and vector control interventions data. FindingsThe developed ddPCR assays demonstrated high accuracy and specificity, with matching Kdr mutation frequencies between pooled samples and individual genotyping. The pilot surveillance study confirmed the low prevalence of Kdr V1016G and I1532T mutations in most French populations, though some populations exhibited a moderate decrease in susceptibility to deltamethrin. Deltamethrin spraying intensity was weakly correlated with Kdr I1532T mutation frequency while no correlation was observed with deltamethrin susceptibility, suggesting that insecticide selective pressure from curative vector control activities is at most a minor driver of resistance. ConclusionThese novel ddPCR assays provide a simple, cost-effective and high-throughput method for quantifying the frequency of Kdr mutations in Ae. albopictus populations. Their implementation in routine country-wide surveillance programs will enhance the detection of emerging resistance, and inform vector control strategies, preventing arboviral disease transmission. Author summaryAedes albopictus, the Asian tiger mosquito, is an emerging global threat due to its ability to transmit the dengue, chikungunya and Zika viruses among others. Recurrent use of insecticides to prevent infections selects resistant mosquitoes. Kdr (knock-down resistance) mutations confer resistance to pyrethroid insecticides, like deltamethrin. In order to track the arrival of these mutations in a population, individual PCR tests are routinely used, which is very inefficient and costly. The digital droplet PCR tests presented in this study can instead be performed on pools of mosquitoes from the field, greatly increasing the output while reducing the costs. This makes them ideal for use in the surveillance of these mutations in field populations. We first confirmed that the tests are as efficient in pools as they are on single mosquitoes, then we applied these tests in a pilot study on French field mosquitoes to demonstrate their applicability for use in surveillance. Finally, we explored the link between deltamethrin sprayings, bioassay mortality and Kdr mutations frequency, which is non-existent. This means the current management of mosquito populations via insecticide use is sensible. We believe these tests have a place in streamlining the future surveillance programs of these mutations in the field.

Western flower thrips (WFTs) are critical vectors of tomato spotted wilt virus (TSWV), transmitting it via a circulative-propagative cycle. The insect-virus relationship is unusual in that only larvae can acquire the virus for transmission to plants to occur. During the larval stage, the virus circulates and replicates within many organs, reaching the salivary glands before the insect pupates, and remaining in infected organs when the insect becomes an adult. The virus continues to replicate in the salivary glands of adult insects, after which it is inoculated into plants via saliva during feeding. Understanding the interactions between TSWV and the WFT salivary glands is critical to furthering investigations of TSWV inoculation and efforts to block the spread of this devastating plant virus. Here, we document transcriptomic changes associated with TSWV infection of the salivary glands of adults (males and females) and second instar larvae. Gene sets enriched in adult male, female, and larval genes revealed a core set of genes associated with WFT salivary glands, as well as genes that differed between sexes and between adults and larvae. The transcriptome response to TSWV infection was higher in larvae (second instar in this study) than in adults, with nearly a 10x increase in differentially expressed genes. We hypothesize this occurred because larvae efficiently acquire the virus and the virus first enters the SGs at the L2 stage, whereas adult SGs are infected only if acquisition occurred in the larval stage. Thus, assessment of larvae detects responses to the early stages of infection, while assessment of adults detects responses to the later stages of infection. Similarly, functional changes in larval salivary glands were more diverse, with significant transcriptome differences associated with growth and development in this tissue during infection. Lastly, a comparative analysis of changes in a published SG proteome revealed a correlation between transcript and protein levels during infection, but little overlap between significant TSWV-responsive transcripts and proteins. These studies provide critical insight into the molecular changes associated with the first breach of the SGs in larvae by TSWV, revealing a markedly different transcriptomic response compared to that in adults.

Circadian clocks enable mosquitoes to anticipate recurring environmental variations and coordinate behaviors critical for survival and disease transmission, such as locomotion, reproduction, host-seeking, and blood-feeding, with times of day when performance is maximal. In Aedes aegypti, locomotor activity follows a robust diurnal rhythm shaped by endogenous circadian clocks and environmental cues, among which light has been shown to be the primary source of temporal information. While early studies established the role of light in regulating locomotor activity, behavior, oviposition and pupation, it remains unclear which features of a light cycle drive changes in circadian rhythms. This question is increasingly relevant as Ae. aegypti is frequently exposed to artificial and dynamic lighting conditions in urban environments. Here, we investigated how transient changes in light schedules influence circadian rhythms in locomotor activity by systematically manipulating the timing, duration, and direction of light exposure. Using a high-throughput assay, we tested over 1900 individuals, including wild-type and timeless knockout mutants, and showed that a single day of al tered lighting is sufficient to induce robust phase shifts, with no evidence of masking effects. A 6-hour light pulse was sufficient to re-entrain mosquitoes regardless of the timing of the pulse, and phase shifts were primarily driven by the offset time of the light pulse, indicating that light-offset acts as a major zeitgeber. Together, these findings challenge conventional assumptions about the timescale of circadian synchronization and highlight the remarkable plasticity of mosquito behavior in response to anthropogenic light. Eventually, these effects could explain the rapid adaptation of the species to urban environments and have potential consequences for disease transmission dynamics.

Short-chain dehydrogenases/reductases (SDRs) constitute a large enzyme superfamily involved in endogenous metabolism and xenobiotic biotransformation. In the parasitic nematode Haemonchus contortus, SDRs may catalyze the carbonyl reduction of a benzimidazole anthelmintic flubendazole (FLU), whose increased reduction is associated with FLU resistance. This study thus investigated the constitutive expression of SDRs and their inducibility by FLU in drug-susceptible and benzimidazole-resistant strains of H. contortus. The expression of 23 sdr genes was analyzed by quantitative PCR, while targeted proteomic assays enabled the quantification of 15 SDR proteins. In adult nematodes, pronounced sex-dependent differences were detected at both transcript and protein levels. Resistance-associated alterations were less pronounced and were observed predominantly in males, with SDR9, SDR12, SDR15, and SDR20 displaying increased protein abundances in the resistant strain. Exposure to FLU induced only minimal transcriptional responses in juvenile stages, whereas adult nematodes exhibited marked sex- and strain-specific responses in the expression of SDRs. The strongest transcriptional effects were detected in resistant males, while significant protein-level changes following FLU treatment were observed exclusively in adults of the drug-susceptible strain. Notably, SDR9 and SDR20 combined resistance-associated expression patterns with responsiveness to FLU exposure. Taking together, the first targeted proteomic characterization of SDRs in H. contortus revealed several SDR isozymes with constitutive overexpression in resistant nematodes and/or inducibility by FLU, suggesting a potential role in adaptation to anthelmintic exposure.

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.

In vertebrates, top-down visual attention is a cognitive process where internal goals modulate the tuning of peripheral sensory systems. This leads to increased perceived contrast to both goal-relevant objects and areas of the visual field that are attended. Such a system would also be beneficial to bees, enabling them to detect and recognise the most profitable flowers in their environment. We tested whether bumblebees possess a top-down attentional system resembling that seen in vertebrates. We trained two groups of bees to collect rewards under high contrast targets. To potentially induce a difference in attention while searching for the targets, one group received a higher concentration of sucrose rewards compared to the other. During tests, the targets were presented with a series of lower contrasts to measure the contrast sensitivity curves of the bees induced by the different learnt reward levels. We predicted a stronger effect of any attention-like process on contrast sensitivity in the high reward group. We also repeated this experiment with the neonicotinoid pesticide imidacloprid dissolved in the sucrose rewards to test whether this affects bee attention. Across all test contrasts, higher rewards significantly increased bee accuracy when locating targets, lowered contrast thresholds and reduced the latency to make first choices. Imidacloprid reduced bee accuracy but did not influence first choice latency. These results suggest that learnt floral rewards can influence bee behavioural contrast sensitivity in a manner resembling vertebrate top-down attention and that imidacloprid may modulate this through effects on their nervous system.

The vast majority of birds are diurnal and concentrate their vocal activity during daylight hours. However, some diurnal birds can also be vocally active at night, although the functions of this phenomenon remain poorly understood. We conducted playback experiments in the Warta Landscape Park (central Poland) to determine whether nocturnal singing by diurnal birds serves breeding-related functions by analysing responses to playback of songs from unfamiliar conspecific males. The sedge warbler (Acrocephalus schoenobaenus) was selected as the focal species because it exhibits relatively high levels of nocturnal song activity, while nine additional diurnal species detected near focal sedge warbler territories were included to explore whether responsiveness to nocturnal conspecific song extended across a broader taxonomic range. Playback experiments were conducted during the early and late stages of the breeding season and during the early and late parts of the nautical night. Out of 10 species tested, three responded vocally: sedge warbler, Savi s warbler (Locustella luscinioides), and common snipe (Gallinago gallinago). Sedge warblers did not modify song rate and song duration but increased flight activity after nocturnal playback. Savi s warblers and common snipes produced more vocalisations after playback than before in May, a pattern consistent with territorial defence function. General nocturnal vocal activity was higher at the beginning of the season, suggesting that birds motivation to establish territories and form pairs can extend into the night, providing additional benefits. Moreover, the probability of singing by the sedge warbler was higher in the latter part of the night. Our study demonstrates that nocturnal stimulation of foreign male playback of diurnal birds can elicit vocal responses from conspecifics, suggesting that nocturnal singing can occur in the absence of obvious artificial light pollution, but in the case of some species and environmental conditions, it may contribute to nocturnal social communication, especially in the early breeding season.

The convergent lady beetle Hippodamia convergens is widespread in the Americas and considered an important beneficial insect due to its use in pest control. Early work on this species characterized the beetles as freeze-avoidant (freeze-intolerant), suggesting they survive low winter temperatures by physiologically preventing ice formation to temperatures as low as -15{degrees}C. Here, we show that H. convergens can be freeze-tolerant if ice formation occurs at relatively high temperatures. There was 100% survival following inoculative freezing at -0.5{degrees}C and exposure to -3{degrees}C for 20 hours, as well as freezing that spontaneously occurred in fed beetles exposed to -4{degrees}C for 4 hours. Males exposed to 0{degrees}C or -4{degrees}C for 4 hours had similar mating behaviours (latency to first mating, copulation duration) as control beetles exposed to 4{degrees}C, although sample sizes were too small to determine whether freezing itself had an effect on these behaviours. Several putative cryoprotectants were detected in fat body tissue of H. convergens: glycerol, proline, trehalose, and myo-inositol - in order of abundance. Exposure to -3{degrees}C for 20 hours, whether frozen or unfrozen, did not statistically affect cryoprotectant accumulation, although there was a trend towards increased glycerol following freezing. This study is the first to describe inoculative freeze tolerance in a lady beetle and establishes a baseline for future studies that examine the mechanisms underlying this freeze tolerance and the effects of freezing on reproductive behaviour. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=69 SRC="FIGDIR/small/744689v1_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@1c0382org.highwire.dtl.DTLVardef@12a5932org.highwire.dtl.DTLVardef@145f4adorg.highwire.dtl.DTLVardef@1c2cb22_HPS_FORMAT_FIGEXP M_FIG C_FIG O_LIHippodamia convergens can tolerate freezing at high subzero temperatures C_LIO_LIMating behaviour of males is normal after chilling or freezing C_LIO_LICryoprotectant accumulation may support overwintering survival C_LI

The marine biodiversity inhabiting rocky shores in the Colombian Pacific remains largely undocumented, primarily due to geographic isolation, logistical challenges, and socio-political constraints. To address the existing knowledge gap, we conducted an expedition to enhance baseline biodiversity knowledge in rocky shores by integrating multiple complementary approaches, including visual censuses, specimen collection with morphological identification, environmental DNA (eDNA) metabarcoding and DNA barcodes. eDNA samples were collected at four coastal sites adjacent to rocky substrates, along with biological specimens obtained from fourteen locations through SCUBA diving at depths ranging from 1 to 25 meters. Tissue samples were subjected to genomic DNA isolation, followed by the generation and validation of cytochrome c oxidase subunit I (COI) barcode sequences, which were subsequently corroborated through taxonomic assessment to ensure accurate species identification. eDNA metabarcoding analyses yielded over 7 million high-quality sequence reads. Although taxonomic resolution at the species level was constrained by the limited completeness of reference sequence databases, a total of 106 species and 83 families were successfully identified, predominantly within the classes Actinopteri, Chondrichthyes, and marine mammals. From the 769 specimens obtained we generated 871 sequences, including 414 validated COI barcodes representing 76 species across 64 families. The integration of DNA barcoding and eDNA approaches resulted in over 1,400 taxonomic detections spanning five phyla, with only six species shared between methodologies. Richness and diversity varied among sites, and revealed significant differences along the coastline between Jurado and Cupica Gulf. All sequences were deposited in BOLDsystems database under the CCBIO project and were visualized through OBIS and GBIF databases. These findings provide the first molecular-based baseline for rocky shore biodiversity in the Colombian Pacific, highlighting the value of integrative approaches for monitoring and conservation.

Catenulids are free-living flatworms, common in eutrophic freshwaters such as ponds, ditches, or peatbogs, with most of the diversity described from tropical regions to date. Although the majority of the species have been described from warmer climates, most molecular studies have been done on specimens from temperate zones in Europe. We addressed this gap by sampling for exotic species in localities available in a temperate climate. In this study, we investigated catenulid diversity in the greenhouses at the University of Warsaw Botanic Garden and recorded two species known only from tropical areas (Stenostomum paraguayense and Suomina evelinae) and one exotic species recorded previously from a greenhouse in Poland (Stenostomum corderoi). Additionally, in the latter species, we provide evidence for environmentally induced coloration of sensory pits, which has not been reported thus far. We placed the collected species on a phylogeny using barcoding of 18S, 28S, and COI genes and retrieved paraphyly of the family Catenulidae, with S. evelinae forming a sister group to the genus Paracatenula, and hence we propose a revision of its systematic position. In total, we recorded six species, including three with a wide cosmopolitan distribution (C. turgida, S. grande and S. tuberculosum), and provided sequences for five of them, three of which had no previous molecular records (S. paraguayense, S. evelinae and S. corderoi). Thus, we confirm that greenhouses represent an important source of exotic species for taxonomic work on microscopic invertebrates.

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.

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.