Journal of Economic Entomology

Gastropods are a highly diverse and often overlooked taxonomic group of significant ecological and economic importance. Some terrestrial gastropods are critical pests of commercial agriculture and home gardens worldwide. Malacopathogenic nematodes offer an effective biological control method of managing pest slugs and snails as a natural enemy. Pellioditis (syn. Phasmarhabditis) hermaphrodita and Pellioditis (syn. Phasmarhabditis) californica are two species of biocontrol nematodes that have been commercialized, sold as Nemaslug(R) and Nemaslug(R) 2.0 respectively on three continents. Although there is interest in bringing Nemaslug(R) products to the US, they are currently not permitted due to limited knowledge on their North American distribution and effects on non-target and native species. In this study, we investigated the impact of P. hermaphrodita and P. californica on Ariolimax columbianus across two slug-host life stages, in laboratory infectivity assays. The objectives were to 1. determine whether P. hermaphrodita and P. californica nematodes impact survival of A. columbianus, and 2. evaluate whether there are differential effects on survival in juvenile and adult life stages of A. columbianus, in laboratory infectivity trials. We found that P. hermaphrodita caused significant mortality in A. columbianus with 100% mortality observed in both juvenile and adult slug hosts. The P. californica treatment had significant effects on the juvenile A. columbianus group only, with 80% mortality. By contrast, only 16% of unexposed control juveniles and 4% of control adult slugs died during the experiment. These results indicate that P. hermaphrodita and P. californica are lethal to the native, non-target Pacific banana slug (A. columbianus) under laboratory conditions, with mortality differing between juvenile and adult host life stages. Given the ecological importance of A. columbianus, these findings raise concerns for potential non-target effects of P. californica and P. hermaphrodita on terrestrial gastropod communities and emphasize the need for testing biocontrol agents against multiple life stages.

Plant-gall wasp systems provide unique models for studying multitrophic interactions and unique developmental trajectories, yet standardized laboratory protocols for maintaining wild rose hosts (Rosa spp.) and sustaining gall inducers (Diplolepis spp.) are lacking. We developed and tested a method for growing and maintaining translocated individuals of Rosa canina, R. rubiginosa, R. spinosissima, R. gallica, R. tomentosa, and R. pendulina under laboratory conditions over three consecutive years (2023-2026). The goal was to have a constant supply of plant host material for reliably producing galls of D. rosae and D. mayri for experimental use. The protocol integrates soil and substrate composition, photoperiod and humidity regimes, pruning, dormancy management, and controlled exposure to gall-inducing wasps. More than 75% of rose individuals survived the full 3-year period, with consistent annual gall induction across some of the species. This work represents the first reproducible laboratory method for long-term maintenance of wild rose hosts and controlled gall induction by Diplolepis species, while also providing a transferable framework for maintaining perennial woody hosts and experimentally manipulating specialized plant-insect interactions under laboratory conditions, thereby providing a platform for ecological, physiological, and evolutionary studies on these interactions.

Chia (Salvia hispanica L.) is an emerging crop in Bangladesh valued for its medicinal properties and economic significance. In March 2024, target spot-like symptoms were observed in an experimental chia field (24.75{degrees} N, 90.50{degrees} E) at Bangladesh Agricultural University in Mymensingh, Bangladesh with disease incidence ranging from 23% to 47% across approximately 0.25 ha. Initially appearing as brick-red spots, these symptoms developed into target-shaped concentric rings, affecting leaves, stems, and inflorescences. A total of 24 fungal isolates were recovered from infected tissue; two representative isolates (BGECh-3 and BGECh-4) were randomly selected for details characterization. Pathogen identity was established through morphological traits, multilocus phylogenetic analysis of internal transcribed spacer (ITS) and elongation factor 1-alpha (EF-1) genes sequence, and pathogenicity confirmation through Kochs postulates, collectively identifying the causal agent as Corynespora cassiicola. The isolates demonstrated a broad host range, successfully infecting brinjal, chili, bottle gourd, country bean, tomato, and soybean. In vitro fungicide sensitivity assays with seven commercial fungicides showed that both isolates were highly sensitive to Goldzim (50% carbendazim), which completely inhibited mycelial growth at 10 {micro}g mL-{superscript 1}. Conza (10% Hexaconazole) and Amister top (18.2% azoxystrobin + 11.4% difenoconazole) reduced growth by up to 85% and 67%, respectively at equal concentration. Other fungicides showed comparatively lower efficacy even at higher concentrations. This study represents the first report of target spot disease of chia caused by C. cassiicola in Bangladesh and provides insights for effective disease management strategies.

Urban pollinator gardens can provide refugia and support diverse populations of native bees amid threats from habitat destruction, pesticides, and potential ecological pressures from the introduced honey bee (Apis mellifera (Linnaeus, 1748)). The University of California, Berkeley, maintained a native bee garden at the Oxford Tract research facility to study the biodiversity, phenology, and foraging habits of urban bees from 2003 to 2009. That garden was decommissioned, and a new garden was re-established in 2019. Using diversity observations from the early 2000s garden and non-lethal sampling techniques, we characterized plant-pollinator interactions between flowers and urban bees in the newer bee garden with a bipartite interaction network. Across 12 flower species, we observed two non-native pollinators, the honey bee (A. mellifera) and the alfalfa leafcutter (Megachile rotundata (Fabricius, 1793)), along with at least ten native bee species across three families (Apidae, Halictidae, Megachilidae). We found that, despite the garden being created for native bees, honey bees accounted for 84% of all pollination interactions. The most abundant native bees were sweat bees (Family: Halictidae). Generalist interactions dominated the network, as both honey and sweat bees foraged on most available flowers. Honey bees showed a significant positive correlation with floral abundance, visiting flowers with the highest number of inflorescences, whereas native bees did not show this preference. These results indicate that native bee garden stewardship could benefit from greater floral diversity, while avoiding the dominance of any single species with high floral abundance, thereby reducing the likelihood of direct competition with honey bees.

Background: The lifespan of insecticide-treated nets (ITNs) varies widely across settings, reflecting both intrinsic product characteristics and external factors related to use, care, and environment. While the resistance to damage (RD) score captures intrinsic product durability, there is no standardized metric to quantify contextual risks. This study presents a proof of concept for the Risk Index (RI), a composite measure of site-level risk factors for ITN physical durability and survival. Methods: We conducted a secondary analysis of durability monitoring data from 44 sites across 15 countries in sub-Saharan Africa, covering 14 ITN products. The RI was calculated as a weighted composite of 12 indicators spanning net handling, net care attitudes, and use environment. Associations between RI and median ITN survival were assessed using weighted linear regression and multivariable mixed-effects models adjusting for RD score, with country included as a random effect. Results: RI scores ranged from 25.1 to 83.7 across study sites. In bivariable analysis, a 10-point decrease in RI was associated with a 4.0-month increase in median ITN survival (95% CI: 1.7-6.3; p=0.001). In multivariable analysis adjusting for RD, this association remained significant but attenuated to 2.2 months (95% CI: 0.1-4.2; p=0.037). Independently, a 10-point increase in RD score was associated with a 3.5-month increase in survival (95% CI: 1.3-5.7; p=0.001). No interaction was observed between RI and RD. Predicted survival differed by approximately one year between the lowest- and highest-risk settings. Conclusion: The RI provides a standardized measure of contextual risk factors affecting ITN lifespan, independent of ITN product type. When used alongside a product's RD score, the RI enables improved interpretation of expected site-level variation in net performance. This combined framework offers a practical basis for incorporating behavioural and environmental risk into vector control planning and for tailoring ITN strategies to local conditions.

BackgroundControl of Aedes aegypti, the primary vector of dengue and other Aedes-borne viruses, is challenged by insecticide resistance, limited efficacy of existing tools and the large and widespread epidemics. Targeted Indoor Residual Spraying (TIRS), a modification of traditional indoor residual spraying focused on Ae. aegypti resting sites, has demonstrated promising results, yet its indirect community-wide effects remain underexplored. Methodology/Principal FindingsWe conducted an entomological cluster-randomized controlled trial in Iquitos, Peru, to evaluate the direct and indirect entomological impacts of TIRS using pirimiphos-methyl. Thirty clusters were randomized to receive either TIRS (15 clusters, 898 structures) or standard Ministry of Health vector control activities (15 clusters, 1,018 structures). Aedes aegypti indoor densities were assessed in the 45 days pre-intervention and at four time points up to 255 days post-intervention using Prokopack aspiration. Generalized linear mixed models with a negative binomial link were used to estimate incidence rate ratios (IRRs) and calculate efficacy (1-IRR) for houses that received TIRS (direct effect) and untreated houses in TIRS clusters (indirect effect). Direct efficacy reached 96% at 15 days post-spraying and remained significant (40%) at 255 days post-spraying. Indirect efficacy reached 69% at 15 days and declined to 7% by 255 days post-spraying. Despite only 57% household-level TIRS coverage, both direct and indirect impacts on Ae. aegypti were significant during early post-intervention surveys, and after 8 months in TIRS clusters. Conclusions/SignificanceTIRS provided substantial and sustained reductions in indoor Ae. aegypti density, including measurable indirect effects in untreated homes within intervention clusters. These findings demonstrate the entomological value of TIRS even at moderate coverage levels and highlight its potential for both preventive and reactive vector control programs and should be considered for implementation by Ministries of Health in dengue-endemic urban settings as well as by the U.S. military when deployed to tropical or subtropical locations.

Avian collision rates are certain to rise as renewable energy industries roll out wind and solar farms to reduce fossil fuel impacts in biologically diverse areas of the world. Technological solutions are often sought to decrease mortality rates, but for developing nations automated shut downs are expensive, and alternatives required. A promising route is to increase blade visibility to birds using high contrast colours. Despite the success of the solid black-blade experiment in Norway only one other black-blade field-study in the Netherlands has explored this possibility, with no significant results. We tested the use of colour-patterned blades at a species-rich, 37-turbine, wind facility in Hopefield, South Africa. Two broad "signal red" stripes were applied to a single blade at four high-fatality turbines, in 2023 by Umoya Energy. Avian fatality rates were compared before and after painting using the Before-After-Control-Impact (BACI) approach. Seventy-five fatalities of 23 species of raptors, passerines and wetland species over 24 months were compared for the same 20 turbines after patterning with two sets of controls: (i) their four nearest neighbours (NN) and (ii) all 16 controls (AC). Over 32 months 25 fatalities were recorded, 23 occurred at the controls and only two at the patterned turbines. Testing with Bayesian Generalized Linear Models (BGLMs) revealed a median 83% reduction in fatalities at the patterned blades for both the NN turbines (credible intervals 14% - 98%) and the AC comparisons (30% - 97%). Bayes Factors (BF) revealed strong statistical support for NN (BF = 49.9) and AC comparisons (BF = 159). There was little evidence that birds avoiding patterned turbines increased fatalities at the neighbouring turbines as there was a small median 15% increase in fatality rates when NN controls were compared with other controls, and weak statistical support (BF = 0.15). Among 14 raptor species recorded on site, 10 species have suffered fatalities. Of seven individuals killed prior to treatment at the four patterned blades, only one was killed post-treatment suggesting blade patterning is equally effective at reducing raptor fatalities. Our results show that patterned blades had a high probability (83%) of reducing fatalities with strong statistical support despite the small samples. This supports the Norway experiment in a high diversity African setting, but with red patterns not a solid black design. The strong effect of red stripes may arise from both the high contrast it provides and the possible warning effect that red may elicit. We call for additional experiments to differentiate the effect of patterns and colours for the optimal design to reduce avian-turbine collisions.

Citrus yellow vein clearing virus (CYVCV) is the causal agent of an emerging disease representing a potentially high-impact threat for citrus production. Despite remaining outside Europe for decades, CYVCV has now expanded towards two important European citrus producers, Italy and, more recently, Spain. The presence of this virus in the EPPO region represents a current threat with unpredictable and potentially devastating consequences for European citriculture. Therefore, urgent protective measures need to be taken to prevent CYVCV spread and minimize its impact. Diagnostics is a key measure in the management of viral diseases, highlighting the need for harmonized methods suitable for reliable routine detection of the currently known CYVCV diversity. In this study, an inclusive, efficient and highly sensitive real-time RT-qPCR for the detection of CYVCV in plant material and transmission vectors has been developed and validated according to EPPO standards. Moreover, the validated method has been successfully adapted to both PCR digital platforms, that allow high-sensitive absolute quantitative detection, essential in the diagnostics at low viral concentrations; and PCR portable tools, that can be applied in a real diagnostic context for on-site detection. This versatility combines standard validated performance, absolute sensitive quantitation and real on-site detection. The study has also addressed sampling strategies to support reliable molecular diagnostic performance. Our results represent an improvement in the detection of CYVCV to be applied in epidemiological studies and different real diagnostic contexts for the containment of this important citrus pathogen.

Prosopis juliflora is an invasive alien plant species and a problematic weed that poses significant ecological and socio-economic challenges in Ethiopia, particularly in the Afar rangelands. The study explored the diversity and effects of insect herbivores communities feeding on the flowers and pods of P. juliflora to determine their role in limiting reproductive success across three selected ecological sites: Amibara, Gewanne, and Aysayita. A total of 118 adult insect specimens were collected between January and November 2021 using a sweep net and hand collection methods. Community structure, analysis via the Shannon Wiener diversity index, strongly influenced damage pattern. Amibara exhibited the highest insect diversity resulting in significant reproductive damage, including 5.98% of flower loss and 10.39% pods tunneling, primarily caused by Chrysomelidae and Pyralidae. Conversely, Gewanne was showed lower diversity, but higher sap-sucking (13.39 % shriveled pods; 5.11 % flower curling) were caused by Aphididae. Overall, 18.41 % of the pods, and 11.59 % of the flowers were exhibited insect related injury. These finding confirm that more internal seed predation and nutrient depletion were revealed significantly reduce viable seed production. The result was suggested that natural insect communities currently function as partial biological control agents. This indicates strong potential for developing integrated biological control strategies to manage P. juliflora invasion in Ethiopia rangelands.

Obtaining tomato plants with firm and intact fruit is one of the main goals in tomato breeding programs. Achieving these goals through conventional breeding is time-consuming and can lead to the loss of unwanted traits. In other hand, consumers are concerned about the presence of transgenic elements in plants acquired through RNA interference. The use of CRISPR/Cas9 technology has made it possible to overcome the above-mentioned shortcomings. In this study, the {beta}-D-N-acetylhexosaminidase ({beta}-hex) gene, which is involved in tomato fruit ripening, was knocked out using CRISPR/Cas9. In the resulting mutant plant genome, an indel mutation was found in exons 1 and 2 of the {beta}-hex gene. Plants with a mutation in their genome were observed to have increased fruit firmness and shelf life compared to control plants without affecting fruit quality.

IntroductionThe environmental impacts of pesticides have raised increasing concern, prompting the development of indicators to assess associated risks across ecosystems. Two main categories are generally distinguished: score-based indicators, which aggregate variables into scores, and risk-based indicators, grounded in the definition of risk as the product of hazard and exposure. Although more data-intensive and more complex to implement, risk-based indicators are recognized to better preserve proportionality with actual risk levels. ObjectivesThis study presents Phytorisque, a model based on the exposure-toxicity ratio to monitor risks associated with pesticide use in Walloon agriculture, from farm to regional scales, and to identify the most contributing active substances in support of risk-reduction policies MethodPhytorisque is a hybrid model that combines mechanistic, empirical, and statistical approaches, integrating quantities of active substances, their ecotoxicological characteristics, and their mobility, persistence, and bioaccumulation properties to generate indices specific to different environmental compartments. ResultsThe indices obtained enable comparison across substances, agricultural sectors, years, and management scenarios. The Phytorisque model provides an integrated assessment of risk across environmental compartments. It can monitor risk evolution over the years for policy impacts evaluation, diagnose the most problematic substances and prospect environmental risks associated with the use of chemical phytoproducts. ConclusionsPhytorisque provides an integrated risk assessment approach adapted to temporal monitoring, diagnosis, and forecasting. It is a relevant operational tool for supporting regional strategies aimed at reducing pesticide-related risks. The model is also transferable to other regions through the adaptation of parameters to local conditions and context.

Giraffes, unlike other large mammals, typically pose minimal risk to humans, their animals, and crops, so they are traditionally not involved in human-wildlife conflict. Tree crops, however, are expanding across Africa, resulting in crop raiding by giraffes and retaliatory snaring, poaching, and killing of giraffes in response. The dynamics of giraffe crop raiding, however, are poorly understood, making effective interventions difficult to implement. To better understand key factors for humans and giraffes that mediate crop raiding, we used a multi-method approach to estimate giraffe abundance and activity, understand farmers perceptions and decisions, and test countermeasures around Garissa Giraffe Sanctuary in eastern Kenya. We hypothesized that 1) giraffe farm invasion would occur in dry seasons, 2) farms growing mangoes would be more likely to be invaded, 3) reducing invasion with only physical barriers would be less effective than adding behavior-based countermeasures, 4) perceptions would match giraffe activity and 5) countermeasure adoption would be driven by cost. We found that invasion and crop raiding primarily occur during the dry season and are associated with mangoes. Farmers are using many countermeasures. Effective countermeasures target giraffe behavior combined with physical barriers. Countermeasures are most effective when negative associations with humans are reinforced. Floodlights and speakers that play predator calls both reduce invasion, but only if used consistently. Overall, farmers perceptions matched giraffe dynamics. Availability was the most important factor in farmers willingness to try a countermeasure. Our results suggest that conflict can be reduced and there is interest from farmers in doing so, but use of countermeasures should be consistently applied and supported by making necessary equipment and instructions available.

Background and AimsPlants deploy triterpenoid saponins as chemical defences against herbivores, yet it remains unclear whether insect digestion detoxifies these compounds or generates equally or more active metabolites. Because saponin bioactivity depends strongly on glycosylation patterns, we examined the fate and defensive activity of hederagenin-derived saponins during herbivory. MethodsLarvae of Plutella xylostella were fed leaf discs containing structurally defined hederagenin-derived saponins. Saponin composition in treated leaves and larval frass was analysed by LC- qTOF-ESI-MS/MS. Feeding assays were used to compare the antifeedant activity of mono- and bidesmosidic forms. Key ResultsLarvae selectively metabolized complex hederagenin-derived saponins into simpler forms, with cellobiosides converted into monoglucosides during digestion, resulting in a marked shift in saponin composition between ingested material and frass. Feeding assays showed that monodesmosidic saponins strongly deterrer feeding, whereas bidesmosidic saponins were largely inactive. The loss of activity in bidesmosidic saponins was not explained by differential metabolism, indicating that glycosylation patterns directly determine biological function. ConclusionsInsect herbivores selectively modify saponin structures through deglycosylation, thereby altering their defensive properties. Our findings demonstrate that glycosylation governs both saponin activity and metabolic fate, highlighting insect-driven turnover as a critical component of plant chemical defence during plant-herbivore interactions. Issue SectionOriginal article

The high efficiency of genome editing presents a challenge when modifying genes associated with viability, welfare, or fertility issues, as implementation of the technology frequently results in mosaic animals with bi-allelic mutations. Combining deactivated Cas9 (dCas9) with Cas9 has been proposed as a strategy to protect one of the two target alleles from editing. We piloted this strategy with 11 genes that are reported as homozygous lethal or associated with welfare issues. We showed that the viability of founders was significantly increased when using 80:20 or 90:10 dCas9:Cas9 ratios, whereas the 70:30 ratio did not yield an equivalent protective effect. The associated overall production rate of mutated founder per manipulated embryo was significantly higher for the 80:20 ratio. Concomitantly, an increased proportion of dCas9 was associated with a significant increase in retention of unedited target alleles but, importantly, did not hinder germline transmission. In addition, editing genes in a paralog cluster with a combination of dCas9 and Cas9 reduced unwanted off-target editing, illustrating a further potential applicability of this approach. This study defines the optimal ratio between dCas9 and Cas9 for strategies aimed at achieving mono-allelic mutations within mosaic founders and proposes a means to reduce the incidence of off-target effects in experiments with limited gRNA options.

Sex ratio is a key demographic parameter shaping population dynamics and evolutionary trajectories. In biocontrol agents, demographic bottlenecks during species introduction to a new habitat and subsequent mass rearing can elevate inbreeding, potentially biasing sex ratios through sex-specific mortality associated with inbreeding depression. Moreover, reproductive endosymbionts such as Wolbachia are known to manipulate host reproduction and further skew sex ratios. However, the relative contributions of these processes to sex-ratio variation remain poorly resolved. In this study, we evaluated the effects of cross-generational full-sibling inbreeding and Wolbachia infection on sex ratio and key life-history traits in the biocontrol beetle Zygogramma bicolorata using controlled laboratory crosses across three generations. Inbreeding did not significantly alter offspring sex ratio, which remained close to parity across generations, while pupal mortality increased in later generations, consistent with delayed expression of inbreeding depression. Adult body weight remained largely unaffected by inbreeding. Wolbachia infection was detected in a subset of females and was associated with a modest but significant increase in female-biased offspring production, although the effect was variable across lineages. Strain typing identified a single supergroup A Wolbachia, consistent with previous descriptions of the wBic strain from this species. These findings indicate that sex-ratio variation in introduced populations of Z. bicolorata is not driven by inbreeding alone but instead emerges from the interaction between demographic processes and symbiont-mediated effects, providing crucial insights for optimizing biocontrol programs where sex-ratio stability is essential for population establishment and persistence. SignificanceSex ratio is a key determinant of population growth and stability - the essential parameters determining success of biocontrol programs. Yet, the mechanisms shaping sex-ratio variation remain poorly resolved. Using controlled crosses in Zygogramma bicolorata, we show that short-term inbreeding does not directly alter sex allocation, despite inducing delayed fitness costs through increased pupal mortality. In contrast, Wolbachia infection contributes to female-biased offspring production, although with variable outcome across lineages. These findings demonstrate that sex-ratio variation in Z. bicolorata arises from the interaction of demographic processes and symbiont effects, rather than a single mechanism, with important implications for predicting the establishment, persistence, and efficacy of mass-reared biocontrol populations.

QuestionsEcological monitoring, repeated collection of ecological data, is essential to document how ecosystems respond to change. In grasslands, different vegetation monitoring protocols are used across disciplines, making it difficult to address multiple management objectives or research questions. We asked four questions about how three common vegetation monitoring protocols compare. (1) How do the protocols differ in how they collect data? (2) How do the protocols differ in their utility? (3) In what ways do vegetation measurements quantitatively differ across protocols? (4) What are each protocols strengths? LocationThis study was conducted on working ranches in the Southern Great Plains with vegetation consisting mainly of native forbs and grasses. MethodsWe implemented three protocols at each site: (1) the Rangeland Analysis Platform (RAP), (2) the Grassland Effectiveness Monitoring (GEM) protocol, and (3) a typical pollinator ecology survey protocol. We qualitatively compared each protocols utility and quantitatively compared cover measurements that each produced. ResultsAll three protocols displayed positive associations within cover categories, but differed in actual cover measurements. The RAP protocol, which uses remote sensing, measured the highest total vegetation cover. The GEM protocol, a line-point intercept method, had more capability to capture fine-scale cover patterns. The GEM protocol measured the most bare ground while the Pollinator protocol measured more forb coverage. ConclusionFine-scale methods like the GEM protocol are most appropriate to address objectives that require capturing small patterns that would otherwise be overlooked with methods like quadrats or remote sensing. Remote sensing is advantageous when monitoring large areas or inaccessible land, but may over-estimate cover. The Pollinator protocol is best equipped to address questions regarding flower abundance and richness. Similarities among protocols can facilitate synergy across disciplines for more effective monitoring. We emphasize the importance of denoting a clear scale and scope of monitoring objectives before selecting methods.

Over the past decade, the global rise in invasive species has accelerated at an unprecedented rate, intensifying threats to ecosystems, human health, and economies worldwide. Newly invasive taxa, such as Tropilaelaps mites, are of particular concern for apiculture and agroecosystems. Despite growing concern about the spread of Tropilaelaps mites and other arthropods, limited resources are available to assess their invasive potential. We characterized 118 invasive arthropod species using available literature to identify key biological and ecological traits associated with invasive potential. We developed predictive generalized linear mixed models (GLMMs) to determine the traits most important for predicting invasive potential (number of invaded regions), and the top-performing models were subsequently applied to Tropilaelaps mercedesae. Several traits were identified as significant predictors of invasiveness, including the degree of human association, resilience at small population sizes, diet breadth, maximum annual number of generations, altitude range, and the interaction between human association and temperature range. Notably, T. mercedesae was predicted to be capable of invading 160 regions, ranking it within the top 10% most invasive species among those evaluated (12th out of 119), ranked just below the cosmopolitan Varroa destructor mite. These findings position T. mercedesae as a high-risk, yet under-recognized, invasive threat. Collectively, this demonstrates the power of predictive trait-based modeling to inform invasion risk prior to widespread establishment and underscores the urgency of reallocating resources toward surveillance, research, and proactive management strategies rather than relying on costly, often ineffective post-establishment eradication.

Giant pangolin (Smutsia gigantea) is one of the least studied pangolin species worldwide, with no published hematological and biochemical data available. We report the first blood parameters from a rehabilitated adult male from Campo Maan National Park (southern Cameroon). Hematological and biochemical findings are described and discussed in relation to available data from other pangolin species. These preliminary results provide the first reference framework for this species and highlight their relevance for clinical assessment, health monitoring, and conservation management.

The sunflower sea star (Pycnopodia helianthoides) suffered a catastrophic population decline across its range from 2013 to 2017 due to the devastating Vibrio pectenicida FHCF-3 driven sea star wasting disease (SSWD) pandemic with minimal signs of population recovery. The functional extinction of this apex predator across substantial parts of its range has created a need to identify and track the remaining intact populations. Environmental DNA (eDNA) approaches provide a simple, cost-effective, and non-destructive method for monitoring occurrences, and in some cases abundances, of marine species, consistently outperforming visual occurrence monitoring efforts in sensitivity, speed, and cost. Here, we designed, developed, and validated a P. helianthoides-specific eDNA assay to identify refugia, using both quantitative and digital droplet PCR approaches. We first generated the most comprehensive sea star mitochondrial genome reference database to date (n=93 taxa, n= 15 novel). We then used unikseq and Geneious bioinformatics software to identify the unique nad5 gene region and design a highly specific hydrolysis probe-based PCR assay. We validated the performance of this assay through laboratory, mesocosm, and field testing, demonstrating a highly specific and sensitive assay. In a field application of the new assay across regions in British Columbia, Canada, we found a positive correlation between P. helianthoides eDNA concentrations and biomass density, especially when appropriately accounting for spatiotemporal integration scales (R2=0.67). The eDNA assay provides a rapid and scalable tool for monitoring the sunflower sea star which has been proposed for listing as threatened under the U.S. Endangered Species Act of 1973. Molecular tools like the one presented here enhance management and recovery efforts not only by identification and monitoring of remnant wild populations, but also by helping to assess population level response and recovery following reintroduction efforts.

Passive acoustic monitoring poses an immense potential to assess avian diversity in many habitats, including agricultural landscapes. At the same time, automated recorders generate large datasets which present a challenge for processing and effectively assessing biodiversity. Methods such as manual listening by experts, automated detection algorithms like BirdNET and calculating acoustic indices all present different trade-offs in assessment of biodiversity through passive acoustic monitoring. In the present study we recorded soundscapes in a low-intensity agricultural landscape in western Turkiye in all four seasons. Two expert ornithologists listened to a subset of these recordings identifying bird species from the recordings. We also ran the same sample of recordings on BirdNET to compare BirdNET detections with expert detections and calculated acoustic indices for each recording. The results showed that BirdNET detected more species than experts, although some may not be reliable detections. Two acoustic indices (bioacoustic index and acoustic complexity index) were correlated positively with number of species detected by experts and one (normalized difference soundscape index) with number of species detected by BirdNET but the correlations were modest. The results show that acoustic indices may have limited value in detecting biodiversity and automated detection algorithms may do a better job, although these may need to be trained with local data to improve detection and classification.