Ecology and Evolution

The coleoid cephalopods (octopus, cuttlefish, and squid) are emerging model organisms for neuroscience, development, and evolutionary biology. Determining their sex early in life is critical for population management and controlled experiments. Here, we present a protocol to non-invasively determine the sex of multiple cephalopod species as young as 3 hours post-hatching using a skin swab and quantitative PCR (qPCR). We describe steps for designing qPCR primers, swabbing live animals, extracting DNA, running the qPCR, and analyzing the results. For complete details on the use and execution of this protocol, please refer to Rubino et al.1 HighlightsO_LISwab live cephalopods as early as 3 hours post-hatching C_LIO_LIExtract DNA from cephalopod skin swabs C_LIO_LIPerform qPCR-based sex determination C_LIO_LIDesign and validate qPCR primers for new species C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=190 SRC="FIGDIR/small/715692v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@3aa68dorg.highwire.dtl.DTLVardef@8c7e61org.highwire.dtl.DTLVardef@1bd45d9org.highwire.dtl.DTLVardef@134cc4d_HPS_FORMAT_FIGEXP M_FIG C_FIG

Changing climates are reshaping animal populations, but our understanding of how demographic trends are shaped by individual responses to local environmental conditions is often limited to long-term studies with restricted spatial scales. Increasing evidence suggests that climatic extremes exert differential selection pressures across environments, often leading to nonstationary biological responses among populations. Participatory science (i.e. citizen science) observations can detect this variation at large geographic scales, but analyses of these data often lack insight into the individual-level responses that are required to explain the origins of such variation. Here we present a new research framework that uses long-term data to validate, then inform analyses of participatory science data to measure reproductive responses to environmental variation across large geographic scales. We use this approach to investigate how reproduction in a montane-adapted songbird, the mountain chickadee (Poecile gambeli), varies across elevations and latitudes in response to extreme scarcity and extreme accumulation of snow throughout the Sierra Nevada Mountains in North America. Chickadee reproduction in lower and higher elevation populations was often differentially impacted by drought and deluge snowfall extremes, but these relationships varied across latitudes. Reproductive performance in the northern Sierra Nevada was negatively affected by snow deluge conditions at high elevations, whereas snow drought conditions reduced reproductive output at low elevations. These relationships changed in the central Sierras where drought conditions negatively impacted both elevations, but deluge conditions improved reproductive performance at both low and high elevations. Reproduction in the southern Sierra Nevada was less affected by spring snow levels, likely due to the lower snow accumulation and earlier snowmelt in this region. These results emphasize the power of long-term studies to inform and interpret participatory science data in order to better understand how animal responses to environmental extremes vary across large geographic scales.

O_LIThe frequency and intensity of heat events is increasing across marine and terrestrial ecosystems. Within the same ecological community, the relative exposure and sensitivity to heat stress may vary considerably among interacting species, like predators and prey. This can be especially true for species that interact at the aquatic-terrestrial interface, as well as for interactions between primarily nocturnal and diurnal species, making it difficult to predict how such communities will respond to habitat warming. C_LIO_LIThermal limit metrics such as CTmax are often assumed to equate with ecological death because such temperatures impair behavioral activity and/or physiological functioning. Prey that are diurnally active can be more frequently exposed to temperatures that approach CTmax compared to their nocturnal predators, which may use thermal refuges during the day. Yet the impacts of daytime heat exposure on nighttime predation risk remain unknown. C_LIO_LIHere, we compared the thermal environment, performance, and heat tolerance between the predatory blue crab, Callinectus sapidus and one of its prey species, the mangrove periwinkle Littoraria anguilifera in a tropical mangrove ecosystem. We examined how exposing prey to heat stress at and below their CTmax affected their capacity to avoid predation in the field at night when predation risk is highest. C_LIO_LIWe found that acute exposure to temperatures near CTmax during the day increased the prey species susceptibility to predation during recovery at night. Although both interacting predator and prey have high thermal tolerance, prey are exposed to conditions that already reach CTmax, suggesting that current extremes in temperatures may already be influencing vulnerability to predation in this ecosystem. C_LIO_LIOur results suggest that differential exposure to sublethal heat stress in diurnal prey relative to their predator, along with the subsequent impact of these exposures on predation risk, will play a role in shaping these interacting as climate warms. C_LI

O_LIPhenological shifts are a major ecological consequence of climate change, yet studies often focus on single life stages meaning that the potential for carryover effects between life stages remains poorly understood. Failing to account for these effects may lead to inaccurate estimates of phenological shifts, with consequences for predicted synchrony among interacting species. This is especially relevant for temperate systems where climate warming is occurring unevenly across the year. C_LIO_LIHere, we investigated how temperature experienced the previous autumn and winter (during the pupal and egg stage) influences spring phenology in the winter moth (Operophtera brumata), a herbivorous insect with distinct life stages. Using 50 years of local climate data to create five experimental temperature regimes, we first quantified phenotypic plasticity in the duration and temporal variability of pupal and egg development. We then examined how timing of adult moth emergence affects timing of offspring hatching. C_LIO_LIWe found divergent effects of temperature on different life stages; pupal development time was shortest at intermediate temperatures while egg development time decreased linearly with increasing temperature. Furthermore, phenological shifts due to the conditions experienced by the mother were carried over to influence the phenology of her offspring. While this carryover effect was partially compensated during subsequent stages, compensation decreased under warming conditions. C_LIO_LIThese results refine our understanding of the sensitivity of the annual cycle of winter moth phenology to variation in temperature with potential implications for population dynamics and interspecific interactions. Overall, our findings highlight the need to consider the impacts of warming across multiple life stages so that carryover effects can be properly accounted for. Doing so will improve predictions of phenological shifts under future climates. C_LI

The temperate rainforests of the Pacific Northwest of North America harbor many endemic taxa whose evolutionary histories have been shaped by major climatic and geologic events. The enigmatic taildropper slugs (genus Prophysaon) are one example, notable for their ability to autonomize their tails to escape predators. Despite extensive work uncovering the evolutionary history of individual lineages, relationships among the nine recognized species of Prophysaon remain poorly understood due to insufficient molecular data. To address this, we collected transcriptomes for six of the nine currently accepted species of Prophysaon. Using these data, we were able to resolve species relationships, calling into question the existing subgeneric classification based on morphology. We also detected undescribed phenotypic diversity within the P. andersonii--P. foliolatum species complex, with molecular data supporting the distinctness of two phenotypically distinct populations from Washington. Finally, our transcriptomic data suggest a moderate role of introgression in shaping the evolutionary history of Prophysaon. Here, we synonymize the subgenus Mimetarion with nominotypical Prophysaon. Future work should further investigate whether the undescribed diversity detected here represents species level differentiation.

High and low tides occur twice a day (every [~]12.4 hours), with the largest tidal ranges during spring tides around new and full moons (every [~]14.765 days). While these lunar cycles are known to influence many animal phenotypes, particularly the reproduction of coastal animals, the genetic basis of lunar-related rhythms remains unclear. Since phenotypic variation is a valuable resource for elucidating such mechanisms, we examined geographic variation in the lunar-regulated mass spawning of the grass puffer (Takifugu alboplumbeus) along the Japanese coast. We found that western populations spawn during the first half of the spring tides, whereas eastern populations spawn during the second half. Furthermore, although spawning typically occurs a few hours before high tide, this timing is restricted to a specific time window that is earlier in the western populations than in the eastern ones. Behavioral analysis of larvae also revealed a shorter free-running circadian period ({tau}) in the western population than in the eastern ones. As differences in {tau} affect individual variation in the timing of physiological functions and behaviors, we hypothesized that differences in {tau} could account for the different time windows and consequently the observed difference in spawning days. Population genomics analysis identified proline-rich transmembrane protein 1-like (prrt1l) as a candidate gene. Expression of prrt1l was observed in the circadian pacemaker suprachiasmatic nucleus, and triple CRISPR F0 knockout of prrt1l shortened the free-running period in larvae. These findings suggest a potential mechanism underlying the geographic variation in lunar-synchronized spawning behavior. HighlightsO_LIThe geographic variation exists in the lunar-regulated spawning of the grass puffer, with differences in spawning dates and times between western and eastern Japan. C_LIO_LIThe free-running period of western populations is shorter than that of eastern populations, which is consistent with their earlier spawning timing. C_LIO_LIPopulation genomics analysis identified prrt1l as a candidate gene harboring population-specific missense mutations, the knockout of which shortens the free-running period. C_LI

Text AbstractIn this study, we characterized the genetic structure and reconstructed the demographic history of cactus wrens (Campylorhynchus brunneicapillus), an endemic species of desert regions of North America, that shows a clear phenotypic and genotypic variation. We evaluated the effects of historical climate change on the structure and population dynamics of desert species using genomic data through genotyping by sequencing (GBS) and applied a population structure analysis (FST and ADMIXTURE), revealing two genetically differentiated groups: one continental and another peninsular in Baja California. Subsequently, we implemented the MSMC2 coalescent model on data divided into autosomal regions and the Z sex chromosome to estimate changes in effective population size (Ne) through evolutionary time. Additionally, we developed ecological niche models (ENMs) projected to the Last Glacial Maximum (LGM), Last Interglacial (LIG), Present times, and Future (2060 - 2080). Results indicate that both populations maintained moderated Nes before the LGM, experienced severe bottlenecks (Ne [~] 102-103), followed by a sustained expansion. However, recovery was limited to the Z chromosome of the peninsular population. These findings reveal how glaciations and interglacials shaped the evolutionary history of desert species and provide genomic evidence of the splitting of C. affinis from C. brunneicapillus. Article summaryThis research examines how climate changes shaped genetic diversity of cactus wrens across North American warm deserts. Using coalescent methods, researchers tracked effective population size changes over 100,000 years, using ecological niche modeling they predicted habitat suitability across climate periods. Results showed that continental and peninsular populations experienced bottlenecks during the Last Glacial Maximum, followed by demographic recovery on warm periods. However, the sex chromosome (Z) revealed male-biased demographic patterns in peninsular populations. Future projections indicated habitat suitability reductions for peninsular populations, highlighting conservation concerns. These findings demonstrate that past climate shaped genetic diversity of cactus wrens.

Parasite infections in wildlife vary across time and space due to interactions among host biology, ecological processes, and climatic variability. Under ongoing climate change, understanding how temperature, precipitation, or humidity influences parasite dynamics is important for predicting shifts in infection patterns and host-parasite interactions. Here, we examine how variation in climatic conditions is associated with helminth infections in a free-living rodent, the bank vole (Myodes glareolus), across 17 years and multiple spatial scales. Using zero-inflated generalised linear models, we quantified the effects of climatic variables on individual parasite burden. Climatic conditions (temperature and humidity or precipitation) affected helminth infections across all analysed scales, though the strength and direction of these effects differed among parasite species and between temporal and spatial scales. In the temporal dataset, parasite load was associated with seasonal variation in weather conditions, whereas in the spatial datasets, infection levels were linked to yearly average climatic variables. The differences reflect species-specific parasites life histories and transmission strategies. Our findings highlight the importance of analysing individual parasite species rather than overall parasite load or aggregated infection indices when assessing the impacts of climatic variation on host-parasite dynamics.

Community science data are increasingly recognized as important resources for biodiversity research, in part because of the spatial and temporal resolution that they afford. While these data are useful for applications such as describing occurrence patterns, tracking movement of migratory animals, and recording phenological events, they can also be probed for "second-order" purposes, such as documenting species interactions. Here, I present a dataset of more than 35,000 annotated interactions between monarch butterfly (Danaus plexippus) larvae and their associated host plants from the community science platform iNaturalist. I document more than 70 unique species of milkweed hosts (Apocynaceae: Asclepiadoideae) used by monarch larvae, including a number of previously undocumented interactions. Monarchs show strong seasonal turnover in the species of host plants used across the migratory cycle, highlighting the importance of early season hosts like Asclepias viridis and A. asperula in eastern North America and A. californica and A. cordifolia in the west. I also demonstrate that non-native horticultural milkweed species have increased the spatial extent of monarch breeding during winter (November - February) by more than 60%, a pattern previously suggested from observational data but not formally quantified until now. To my knowledge, this represents the largest analysis to date of species interactions using unstructured community science data and highlights the value of platforms like iNaturalist for conducting fundamental research in ecology and conservation.

While the ubiquity of parasites is well understood, the extent to which host demographic factors shape parasite prevalence patterns merits further investigation. Using 15 years of breeding data from blue tits (Cyanistes caeruleus) in southern Sweden, spanning a 26-year timeframe, we assessed the roles of host age, sex, and field site on the odds of infection with three avian malaria parasite genera: Haemoproteus, Plasmodium, and Leucocytozoon. Further, we also evaluated the effects of these demographics on the odds of triple-genus coinfections. We found first-year breeders have fewer infections with Haemoproteus and Plasmodium, and fewer triple infections compared to older age classes, suggesting that birds are accumulating infections over time. Leucocytozoon infections are more prevalent in males than in females, and this may be due to sex-specific differences in physiology. The prevalence of malaria parasites and their coinfections also vary between the three sampling sites, indicative of an effect of host breeding habitat, even at a relatively small spatial scale (neighboring sites were separated by <5km). Across all three field sites, prevalence is overall significantly increasing over time. For Haemoproteus, this increase is more pronounced in older birds compared to younger birds. Such temporal changes in age-related infection patterns would not have been apparent without long-term data thus highlighting the importance of long-term studies for informing our understanding of host demographic effects on parasite prevalence.

Animal-associated bacteria (microbiomes) can have important effects on host phenotypes and fitness. Microbiomes can also vary across individuals in ways that depend on host genotype and environment. Temperature is an especially important environmental factor that can affect the microbiome in a way that depends on host genotype and affects organismal fitness. Thermal stress, in particular, can have dramatic effects on animal microbiomes, including dysbiosis and immune dysregulation. However, most previous work on extreme temperature effects has focused on heat stress. To investigate how low temperatures affect the microbiome of a warm-adapted animal, we characterized the bacterial communities associated with house fly (Musca domestica) males raised at cool (18{degrees}C) and warm (29{degrees}C) temperatures. We sampled two distinct genotypes in these experimental flies, each of which is associated with a particular thermal environment (warm or cool). We contrasted our experimental results with the microbiomes we characterized in wild house flies from two collection sites with different large animals present. We found that temperature has a much stronger effect on the house fly microbiome than the host genotype in our experimental flies. Consistent with the strong environmental effects in our experiment, we found that wild house fly microbiomes differed between the two collection sites. Despite these environmental effects on the house fly microbiome, we did not detect evidence for dysbiosis associated with either cool or warm temperatures. We therefore conclude that the environment has more of an effect on the house fly microbiome than host genotype, but dysbiosis does not occur within the temperature range we considered.

Sarita Lake, British Columbia houses a distinctive population of threespine stickleback (Gastrosteus aculeatus L.) with a phenotype characterized by unusually large individuals relative to nearby conspecifics. We tested the hypothesis that members of this population are not isometrically larger but rather exhibit variation in allometric trajectories that reflect changes in developmental timing impacting the developmental-genetic architecture of the phenotype. We used 3D geometric morphometrics to characterize the size and shape of skulls, pectoral girdles and pelvic girdles from a sample of individuals from nearby freshwater and marine populations and compare them to a sample from Sarita Lake. We showed that individuals from the Sarita Lake population are larger in each body region compared to most other populations examined. Further, these individuals have dorsally expanded skulls and relatively robust pelvic armour. We also showed that the relationship between size and shape is differently structured among body regions and is heavily influenced by non-uniform sexually-mediated variation across populations sampled. Our results reflect complex underlying developmental trajectories, and we suggest that the large phenotype observed may be driven by fecundity selection on female size in combination with a limnetic trophic niche and relatively increased predation pressure in Sarita Lake.

Dams can significantly alter natural riverine systems, but their impact on movement across rivers for most terrestrial vertebrates is poorly known. The completion of Glen Canyon and Flaming Gorge dams in Arizona and Utah (southwestern United States) profoundly changed the Colorado and Green Rivers and have altered habitat for many species. The common side-blotched lizard (Uta stansburiana) offers an excellent opportunity to examine the effects of riverine impoundments on migration and gene flow in terrestrial biodiversity. To assess these effects, we collected tissue samples from 241 Uta stansburiana above and below Glen Canyon Dam and on both sides of the Colorado river at three separate study areas. We used eight microsatellite loci to estimate genetic exchange in the context of genetic diversity and structure. One study area below Flaming Gorge Dam and above Glen Canyon Dam has annual periods of warmer water temperatures and lower flows that are closer to pre-dam conditions, whereas two study areas below Glen Canyon Dam have cold water temperatures year-round, and less pronounced seasonal low flow episodes. We predicted that warmer water temperatures above Glen Canyon Dam would promote greater genetic exchange among populations than below the dam. However, we found evidence for low levels of genetic exchange between sites both above and below Glen Canyon Dam, and a moderate amount of exchange at a site below this dam where lizards could conceivably move from one side to the other. Our results imply that 1) the changes in water temperature and hydrology in dam-altered rivers are a barrier for this species even when the distance from the dam is great; and 2) genetic exchange may be dependent on river morphology. These results are relevant to other small vertebrates, particularly ectotherms, that occupy habitat proximal to a dammed river and has implications for the conservation management of impounded river systems.

Rapid changes to weather caused by climate change have a negative effect on much of the worlds animal populations and species. Some populations are more vulnerable than others to the effects of climate change, and individuals are particularly vulnerable during early development. Good embryonic development is important for vertebrate species because this can dictate their breeding success and survival rates, and disruptions to this phase can have far reaching fitness effects that can last into adulthood and beyond. We looked at the impact of weather (ambient temperature, rainfall and wind speed) on the embryonic development of thorn-tailed Rayadito (Aphrastura spinicauda) at two different latitudes in Patagonia, Chile. We measured the heart rate of embryos just before hatching using an egg buddy machine to determine embryonic development. Optimum development of nestlings is important for fledging, so it is essential that embryonic development is successful. We studied two populations. One is situated in a temperate rainforest on the northern border of Patagonia called Pucon which we studied in 2018 and 2019, with mild temperatures (12 degrees Celsius), high rainfall (636ml) and low wind speeds (6.3km/h). The other is in a sub-Antarctic old growth forest in southern Patagonia called Navarino island which we studied in 2018, 2019 and 2023, which is comparatively drier (138ml), colder (8.3 degrees Celsius) and has higher average wind speeds (16.6km/h). We found that embryonic development was better in the south compared to the north, indicated by higher embryo heart rates near hatching in the south. Development of embryos in the northern site was slower when conditions were cold and windy. Development of embryos in the southern site was unaffected by temperature, rainfall or wind speeds. In northern Patagonia, when minimum temperatures were low and wind speeds high during the period encompassing clutch completion, initiation of full incubation and during incubation, have a negative impact on embryonic development. In contrast, when Rayaditos in the southern population experience slow embryonic development, they extend the incubation period allowing embryos more time to develop before hatching. Our study shows that in the north of Patagonia embryonic development declines over years and that Rayaditos seem not to have adapted to dealing with climate change. On the other hand, in the south of Patagonia, embryonic development is unaffected by climatic factors, suggesting that Rayaditos are adapting to climate change through extending their incubation periods, allowing embryos to fully develop before hatching. It appears that Rayaditos in the northern population are not extending their incubation periods and are not adapting to the threats posed by climate change. To our knowledge, this is the first study of its kind to examine embryonic development in the field and to associate this to changing weather patterns whilst highlighting specific days on which development is influenced.

The long-standing misconception that the Galapagos petrel (Pterodroma phaeopygia) and the Hawaiian petrel (Pterodroma sandwichensis) were conspecific masked the severe vulnerability of the Galapagos population. By the time its distinct status was recognized, the Galapagos petrel was already in marked decline, primarily due to invasive predators. Consequently, sustained rodent control programs have been implemented on Santa Cruz Island. An unintentional one-year failure in rodent control provided a rare quasi-experimental opportunity to quantify the demographic consequences of the invasive black rat predator. During this year, hatching success declined by [~]35% and breeding success by [~]40% relative to long-term means (66% and 62%, respectively), representing a substantial reproductive collapse. Fledging success exhibited a comparatively modest decline (from a long-term mean of 94% to 86% in 2017), suggesting stage-specific vulnerability. These results support the hypothesis that invasive black rats primarily affect early reproductive stages through egg predation and predation on small chicks, while older chicks surpass a critical size threshold that reduces susceptibility. Across the remaining managed years, reproductive metrics exhibited great stability, demonstrating the petrels resilience against other environmental or climatic stressors. Our findings provide robust empirical evidence that invasive rodent control is the dominant driver of reproductive success in this endangered seabird. The quasi-experimental failure underscored both the effectiveness and the necessity of continuous predator management, highlighting the severe and immediate consequences of even short-term lapses.

In many tropical areas, seasonal rainfall leads to distinct dry and wet seasons. Many butterflies developing under wet season conditions develop into adults with large ventral eyespots on the wing margins, whereas those developing under dry season conditions have smaller or no eyespots. In greener, wet season habitats, larger eyespots can divert predator attacks toward the wing margins, while reduced eyespot size improves camouflage in the dry leaf litter-dominated habitat during the dry season. However, the dry season is also characterised by higher desiccation stress than the wet season. We hypothesised that larvae developing under dry season conditions develop into adults with higher desiccation tolerance than those reared under wet season conditions. We tested this by rearing larvae of the butterfly Mycalesis mineus under simulated dry and wet season conditions and assaying the desiccation tolerance of the resulting adults. Butterflies reared in dry conditions survived longer under desiccation stress, lost lesser water during pupal-adult metamorphosis, and were heavier than those reared in wet conditions. We also tested the correlation between eyespot size and desiccation tolerance. A negative correlation between the traits would be expected if similar developmental pathways regulate them. Consistent with this expectation, individuals with smaller eyespots had higher desiccation tolerance. Our results demonstrate plasticity in desiccation tolerance, and suggest that predator avoidance and desiccation tolerance traits may share similar developmental pathways.

DNA metabarcoding is a central tool in biodiversity research and monitoring, producing detailed taxa lists with comparatively little time and effort. One of its limitations, however, is the lack of quantitative data on biomass or abundance. This limitation has two main reasons: 1) template copy number variation and 2) primer-induced amplification bias. Many metabarcoding markers are mitochondrial and mitochondrial copy numbers vary in animal tissues, potentially decoupling sequence counts from biomass. Additionally, primer mismatches can lead to taxon-specific amplification biases, for which PCR cycle calibration has been proposed as a solution. To mechanistically study both effects, we constructed mock communities of different arthropod species. We combined digital droplet PCR and COI metabarcoding to quantify relationships between biomass, mitochondrial copy number and metabarcoding reads. Mitochondrial DNA copy numbers per biomass varied strongly within and among the different taxa. Metabarcoding reads did not reflect input mitochondrial DNA copies without a correction. Attempts to correct for amplification bias via PCR cycle calibration failed as read proportions remained stable across cycles. We therefore mathematically derived an approach to estimate relative amplification bias and initial mitochondrial DNA copy numbers in a sample based on a non-exponential amplification bias model and demonstrate its applicability. Still, the detected high variation in mitochondrial copy numbers and derived prerequisites necessary to calculate amplification efficiencies and mitochondrial copy numbers limit the practical application. Our study highlights fundamental constraints of quantitative metabarcoding and underscores the need for additional methodological approaches for quantitative insights while delivering essential conceptual insights.

Radar aeroecology is dedicated to making ecological inference about aerial wildlife from radar-derived information. While producing unique, large-scale datasets describing biological activity in the sky, radar methodologies are largely incapable of relating these to specific species and are thus taxonomically limited. I describe a computational method to increase taxonomic resolution in vertical looking radar data by dividing detected organisms into morphology and movement-based aerial morphotypes. Using the Birdscan MR1 radar target classifier, wing flapping frequency calculation and target size estimation, I demonstrate a nearly 8 fold increase in classification resolution of bird radar data from the Hula Valley Research station, Israel. Furthermore, by relating each species in the regions species pool to its relevant morphotype, I show that most of these newly separated classes are related to small numbers of species (1-10), providing realistic opurtunities to bridge the taxonomy gap in radar data. By using the morphotype approach, radar aeroecologists can start observing and discussing the concept of "Aerodiversity", analogues to widely used biodiversity, a fundamental measure in ecology and conservation sciences. By analitically adressing taxonomy in radar-aeroecology, practitioners will increase the impact and dissemintation of their work and contribute to a better, more complete understanding of the aerial habitat.

Computational workshops are common in evolutionary biology and are used to share discipline-specific tools and skills with researchers. Despite the perceived importance of these workshops, there is no common set of criteria for workshop success, and there are few peer-reviewed studies investigating the efficacy of workshops or assessing the value of particular instructional techniques in this context. Here, we focused on one key element of a successful workshop: its ability to increase participants motivation to use the methods and tools presented during the workshop. We analyzed the goals, perceptions, and future plans of research practitioners engaging in a workshop on phylogenetic methods of historical biogeography using pre- and post-workshop surveys. Overall, the workshop was successful at motivating participants, and survey responses provided insights into participants perceptions of different activities, including "participatory live coding". Apart from this case study, we aim to highlight the importance of developing a common set of workshop goals in collaboration with other workshop stakeholders and the need for specialized, validated tools for assessing the efficacy of computational workshops for researchers.

Understanding how the fear of predation acts as a driver of spatial distribution is fundamental to animal behaviour research, yet this relationship is not wholly understood in primates such as baboons. Olive baboons (Papio anubis) have evolved a diverse range of antipredator strategies that reduce, but do not eliminate, predation risk from the large carnivores they encounter across their broad geographic range. This raises a critical question: does the need to access essential resources outweigh the risk of predation when determining habitat selection? We addressed this question by examining the relative influence of three environmental factors and relative predator abundance on olive baboon occupancy patterns and detection probability in Serengeti National Park, Tanzania. Using data from 225 camera traps deployed by the Snapshot Safari program, we fitted three separate Bayesian occupancy models, each incorporating the same three environmental covariates (terrain ruggedness index, distance to nearest river, and Normalized Difference Vegetation Index, NDVI), together with the relative abundance of one of three principal predators (lion, leopard, or spotted hyena). This approach allowed us to assess whether environmental covariates associated with baboon occupancy remained consistent across different predator contexts. Baboon occupancy strongly increased with terrain ruggedness in all three models and consistently decreased with a greater distance to rivers. Vegetation greenness (NDVI) showed a positive association with baboon occupancy, though credible intervals narrowly overlapped zero. NDVI also showed a strong positive relationship with baboon detection probability. Associations between predator relative abundance and baboon occupancy varied between models: the relative abundance of lions and spotted hyenas showed no strong association with baboon occupancy, whereas the relative abundance of leopards was strongly correlated with baboon occupancy, consistent with shared habitat preferences. Our findings demonstrate that, independent of predator presence, olive baboon spatial distribution in the Serengeti is primarily and consistently associated with resource-related environmental features. This study expands our knowledge on the ecological factors that influence primate occupancy by showing that, for a behaviourally flexible species with diverse antipredator strategies, access to essential resources can outweigh spatial avoidance of predators in a multi-predator landscape.