Seasonality and alternative floral resources affect reproductive success of the alfalfa leafcutting bee, Megachile rotundata.

Background: Managed populations of the alfalfa leafcutting bee (ALCB), Megachile rotundata (F.), are often not sustainable. In addition to numerous mortality factors that contribute to this, the dense bee populations used to maximize alfalfa pollination quickly deplete floral resources available to bees later in the summer. Providing alternative floral resources as alfalfa declines may help to improve ALCB reproduction. Methods: We examined the relationship between floral resource availability and ALCB reproduction and offspring condition via (1) a field study using alfalfa plots with and without late-blooming wildflower strips to supply food beyond alfalfa bloom, and (2) a field-cage study in which we provided bees with alfalfa, wildflowers, or both as food resources. Results: In the field study, bee cell production closely followed alfalfa floral density with an initial peak followed by large declines prior to wildflower bloom. Few bees visited wildflower strips, whose presence or absence was not associated with any measure of bee reproduction. However, we found that female offspring from cells provisioned earlier in the season, when alfalfa predominated as a source of provisions, eclosed with greater body sizes and proportion body lipids relative to total body mass. For bees restricted to cages, the proportion of offspring that survived to adults was highest on pure alfalfa diets. Adding wildflowers to cages with alfalfa did not affect adult offspring production or female offspring body size and lipid content. Furthermore, although similar numbers of adults were produced on wildflowers alone as with alfalfa alone, females eclosed with smaller body sizes and lower proportion body lipids on wildflowers despite the higher protein content we estimated for wildflower pollen. We found no evidence that adding the late-season wildflower species that we chose to plant enhanced ALCB offspring numbers. Our results highlight the importance of considering multiple measures of reproductive success, including offspring body size and lipid stores, when designing and evaluating floral resource management strategies for agroecosystems.

Bumble bee diet breadth increases with local abundance and phenophase duration, not intraspecific variation in body size.

Patterns of abundance across space and time, and intraspecific variation in body size, are two species attributes known to influence diet breadth and the structure of interaction networks. Yet, the relative influence of these attributes on diet breadth is often assumed to be equal among taxonomic groups, and the relationship between intraspecific variation in body size on interaction patterns is frequently neglected. We observed bee-flower interactions in multiple locations across Montana, USA, for two growing seasons and measured spatial and temporal patterns of abundance, along with interspecific and intraspecific variation in body size for prevalent species. We predicted that the association between spatial and temporal patterns of abundance and intraspecific variation in body size, and diet breadth, would be stronger for bumble bee compared to non-bumble bee species, because species with flexible diets and long activity periods can interact with more food items. Bumble bees had higher local abundance, occurred in many local communities, more intraspecific variation in body size, and longer phenophases compared to non-bumble bee species, but only local abundance and phenophase duration had a stronger positive association with the diet breadth of bumble bee compared to non-bumble bee species. Communities with a higher proportion of bumble bees also had higher intraspecific variation in body size at the network-level, and network-level intraspecific variation in body size was positively correlated with diet generalization. Our findings highlight that the association between species attributes and diet breadth changes depending on the taxonomic group, with implications for the structure of interaction networks.

Roadside habitat: Boon or bane for pollinating insects?

Pollinators, which provide vital services to wild ecosystems and agricultural crops, are facing global declines and habitat loss. As undeveloped land becomes increasingly scarce, much focus has been directed recently to roadsides as potential target zones for providing floral resources to pollinators. Roadsides, however, are risky places for pollinators, with threats from vehicle collisions, toxic pollutants, mowing, herbicides, and more. Although these threats have been investigated, most studies have yet to quantify the costs and benefits of roadsides to pollinators and, therefore, do not address whether the costs outweigh the benefits for pollinator populations using roadside habitats. In this article, we address how, when, and under what conditions roadside habitats may benefit or harm pollinators, reviewing existing knowledge and recommending practical questions that managers and policymakers should consider when planning pollinator-focused roadside management.

Predicting plant-pollinator interactions: concepts, methods, and challenges.

Plant-pollinator interactions are ecologically and economically important, and, as a result, their prediction is a crucial theoretical and applied goal for ecologists. Although various analytical methods are available, we still have a limited ability to predict plant-pollinator interactions. The predictive ability of different plant-pollinator interaction models depends on the specific definitions used to conceptualize and quantify species attributes (e.g., morphological traits), sampling effects (e.g., detection probabilities), and data resolution and availability. Progress in the study of plant-pollinator interactions requires conceptual and methodological advances concerning the mechanisms and species attributes governing interactions as well as improved modeling approaches to predict interactions. Current methods to predict plant-pollinator interactions present ample opportunities for improvement and spark new horizons for basic and applied research.

Diversity and distribution of orchid bees (Hymenoptera: Apidae, Euglossini) in Belize.

Background: Orchid bees are abundant and widespread in the Neotropics, where males are important pollinators of orchids they visit to collect fragrant chemicals later used to court females. Assemblages of orchid bees have been intensively surveyed in parts of Central America, but less so in Belize, where we studied them during the late-wet and early-dry seasons of 2015-2020. Methods: Using bottle-traps baited with chemicals known to attract a variety of orchid bee species, we conducted surveys at sites varying in latitude, historical annual precipitation, elevation, and the presence of nearby agricultural activities. Each sample during each survey period consisted of the same number of traps and the same set of chemical baits, their positions randomized along transects. Results: In 86 samples, we collected 24 species in four genera: Euglossa (16 species), Eulaema (3), Eufriesea (3), and Exaerete (2). During our most extensive sampling (December 2016-February 2017), species diversity was not correlated with latitude, precipitation, or elevation; species richness was correlated only with precipitation (positively). However, a canonical correspondence analysis indicated that species composition of assemblages varied across all three environmental gradients, with species like Eufriesea concava, Euglossa imperialis, and Euglossa viridissima most common in the drier north, and Euglossa ignita, Euglossa purpurea, and Eulaema meriana more so in the wetter southeast. Other species, such as Euglossa tridentata and Eulaema cingulata, were common throughout the area sampled. Mean species diversity was higher at sites with agricultural activities than at sites separated from agricultural areas. A Chao1 analysis suggests that other species should yet be found at our sites, a conclusion supported by records from adjacent countries, as well as the fact that we often added new species with repeated surveys of the same sites up through early 2020, and with the use of alternative baits. Additional species may be especially likely if sampling occurs outside of the months/seasons that we have sampled so far.

Characterization Factors to Assess Land Use Impacts on Pollinator Abundance in Life Cycle Assessment.

While wild pollinators play a key role in global food production, their assessment is currently missing from the most commonly used environmental impact assessment method, Life Cycle Assessment (LCA). This is mainly due to constraints in data availability and compatibility with LCA inventories. To target this gap, relative pollinator abundance estimates were obtained with the use of a Delphi assessment, during which 25 experts, covering 16 nationalities and 45 countries of expertise, provided scores for low, typical, and high expected abundance associated with 24 land use categories. Based on these estimates, this study presents a set of globally generic characterization factors (CFs) that allows translating land use into relative impacts to wild pollinator abundance. The associated uncertainty of the CFs is presented along with an illustrative case to demonstrate the applicability in LCA studies. The CFs based on estimates that reached consensus during the Delphi assessment are recommended as readily applicable and allow key differences among land use types to be distinguished. The resulting CFs are proposed as the first step for incorporating pollinator impacts in LCA studies, exemplifying the use of expert elicitation methods as a useful tool to fill data gaps that constrain the characterization of key environmental impacts.

Mycorrhizae influence plant vegetative and floral traits and intraspecific trait variation.

PREMISE: Arbuscular mycorrhizal fungi (AMF) can strongly influence host plant vegetative growth, but less is known about AMF effects on other plant traits, the relative impacts of AMF on vegetative growth versus floral traits, or AMF-induced intraspecific variation in traits. METHODS: In an experimental greenhouse study, we inoculated seven species of wildflowers with six species of AMF in a factorial design. We assessed how the AMF-forb combinations influenced plant survival, vegetative biomass, and floral traits and whether AMF effects on floral traits were similar in magnitude and direction to effects on vegetative biomass. For one forb species, we investigated intraspecific plant trait variation within and across AMF treatments. RESULTS: AMF species varied from negative to positive in their effects on host plants. AMF often had inconsistent effects on vegetative biomass versus floral traits, and therefore, quantifying one or the other may provide a misleading representation of potential AMF effects. AMF treatments generated key variation in plant traits, especially floral traits, with potential consequences for plant-pollinator interactions. Given increased intraspecific trait variation in Linum lewisii plants across AMF species compared to uninoculated individuals or single AMF treatments, local AMF diversity and their host plant associations may scale up to influence community-wide patterns of trait variation and species interactions. CONCLUSIONS: These results have implications for predicting how aboveground communities are affected by belowground communities. Including AMF effects on not just host plant biomass but also functional traits and trait variation will deepen our understanding of community structure and function, including pollination.

The role of timing in intraspecific trait ecology.

Intraspecific trait variation has tremendous importance for species interactions and community composition. A major source of intraspecific trait variation is an organism's developmental stage; however, timing is rarely considered in studies of the ecological effects of intraspecific variation. Here, we examine the role of time in the ecology of intraspecific trait variation, focusing on plants and their interactions with other organisms. Trait variation due to differences in developmental timing has unique features and dynamics, distinguishing it from variation due to genes or the environment. When time is considered in studies of intraspecific trait ecology, the degree of variability in timing within a population becomes a key factor structuring trait-mediated ecological interactions and community processes.

Effects of Temperature and Wildflower Strips on Survival and Macronutrient Stores of the Alfalfa Leafcutting Bee (Hymenoptera: Megachilidae) Under Extended Cold Storage.

Megachile rotundata (F.) is an important pollinator of alfalfa in the United States. Enhancing landscapes with wildflowers is a primary strategy for conserving pollinators and may improve the sustainability of M. rotundata. Changing cold storage temperatures from a traditionally static thermal regime (STR) to a fluctuating thermal regime (FTR) improves overwintering success and extends M. rotundata's shelf life and pollination window. Whether floral resources enhance overwintering survival and/or interact with a thermal regime are unknown. We tested the combined effects of enhancing alfalfa fields with wildflowers and thermal regime on survival and macronutrient stores under extended cold storage (i.e., beyond one season). Megachile rotundata adults were released in alfalfa plots with and without wildflower strips. Completed nests were harvested in September and stored in STR. After a year, cells were randomly assigned to remain in STR for 6 months or in FTR for a year of extended cold storage; emergence rates were observed monthly. Macronutrient levels of emerged females were assessed. FTR improved M. rotundata survival but there was no measurable effect of wildflower strips on overwintering success or nutrient stores. Timing of nest establishment emerged as a key factor: offspring produced late in the season had lower winter survival and dry body mass. Sugars and glycogen stores increased under FTR but not STR. Trehalose levels were similar across treatments. Total lipid stores depleted faster under FTR. While wildflowers did not improve M. rotundata survival, our findings provide mechanistic insight into benefits and potential costs of FTR for this important pollinator.

The effect of natural disturbances on forest biodiversity: an ecological synthesis.

Disturbances alter biodiversity via their specific characteristics, including severity and extent in the landscape, which act at different temporal and spatial scales. Biodiversity response to disturbance also depends on the community characteristics and habitat requirements of species. Untangling the mechanistic interplay of these factors has guided disturbance ecology for decades, generating mixed scientific evidence of biodiversity responses to disturbance. Understanding the impact of natural disturbances on biodiversity is increasingly important due to human-induced changes in natural disturbance regimes. In many areas, major natural forest disturbances, such as wildfires, windstorms, and insect outbreaks, are becoming more frequent, intense, severe, and widespread due to climate change and land-use change. Conversely, the suppression of natural disturbances threatens disturbance-dependent biota. Using a meta-analytic approach, we analysed a global data set (with most sampling concentrated in temperate and boreal secondary forests) of species assemblages of 26 taxonomic groups, including plants, animals, and fungi collected from forests affected by wildfires, windstorms, and insect outbreaks. The overall effect of natural disturbances on α-diversity did not differ significantly from zero, but some taxonomic groups responded positively to disturbance, while others tended to respond negatively. Disturbance was beneficial for taxonomic groups preferring conditions associated with open canopies (e.g. hymenopterans and hoverflies), whereas ground-dwelling groups and/or groups typically associated with shady conditions (e.g. epigeic lichens and mycorrhizal fungi) were more likely to be negatively impacted by disturbance. Across all taxonomic groups, the highest α-diversity in disturbed forest patches occurred under moderate disturbance severity, i.e. with approximately 55% of trees killed by disturbance. We further extended our meta-analysis by applying a unified diversity concept based on Hill numbers to estimate α-diversity changes in different taxonomic groups across a gradient of disturbance severity measured at the stand scale and incorporating other disturbance features. We found that disturbance severity negatively affected diversity for Hill number q = 0 but not for q = 1 and q = 2, indicating that diversity-disturbance relationships are shaped by species relative abundances. Our synthesis of α-diversity was extended by a synthesis of disturbance-induced change in species assemblages, and revealed that disturbance changes the ß-diversity of multiple taxonomic groups, including some groups that were not affected at the α-diversity level (birds and woody plants). Finally, we used mixed rarefaction/extrapolation to estimate biodiversity change as a function of the proportion of forests that were disturbed, i.e. the disturbance extent measured at the landscape scale. The comparison of intact and naturally disturbed forests revealed that both types of forests provide habitat for unique species assemblages, whereas species diversity in the mixture of disturbed and undisturbed forests peaked at intermediate values of disturbance extent in the simulated landscape. Hence, the relationship between α-diversity and disturbance severity in disturbed forest stands was strikingly similar to the relationship between species richness and disturbance extent in a landscape consisting of both disturbed and undisturbed forest habitats. This result suggests that both moderate disturbance severity and moderate disturbance extent support the highest levels of biodiversity in contemporary forest landscapes.

[Evolution of Emergency Department Characteristics in Child and Adolescent Psychiatry: A Retrospective Review over Two Decades]. / Entwicklung der Charakteristika von Notaufnahmen in der Kinder- und Jugendpsychiatrie.

Evolution of Emergency Department Characteristics in Child and Adolescent Psychiatry: A Retrospective Review over Two Decades Abstract. Objective: Emergency inpatient admissions to child and adolescent psychiatric hospitals because of a mental health crisis represent a substantial proportion of all inpatient admissions and have increased substantially over time. This study examines changes in the characteristics of this patient group at a university care clinic over two decades. Method: We evaluated the emergency admissions from 1996, 2002, 2008, and 2014 of the Child and Adolescent Psychiatry Clinic in Tübingen retrospectively using sociodemographic data, psychosocial circumstances, and diagnoses. Results: We evaluated a total of N = 403 emergency admissions. The emergency admissions in the periods mentioned increased by 405 %. Especially patients from families with separated parents and with multiple diagnoses increased over time. Conclusions: From 1996 to 2014, there was a significant increase in emergency admissions. The results also indicate that more complex disease situations and less favorable psychosocial conditions are occurring. The findings underscore the need to improve the clinical care of children and adolescents during acute mental health crises and work toward their prevention. There is also a need to focus broad societal discussion on improving overall mental health during childhood development. There is an urgent need for prospective studies to identify the factors leading to the increase in emergency admissions among children and adolescents.

Widespread vulnerability of flowering plant seed production to pollinator declines.

Despite evidence of pollinator declines from many regions across the globe, the threat this poses to plant populations is not clear because plants can often produce seeds without animal pollinators. Here, we quantify pollinator contribution to seed production by comparing fertility in the presence versus the absence of pollinators for a global dataset of 1174 plant species. We estimate that, without pollinators, a third of flowering plant species would produce no seeds and half would suffer an 80% or more reduction in fertility. Pollinator contribution to plant reproduction is higher in plants with tree growth form, multiple reproductive episodes, more specialized pollination systems, and tropical distributions, making these groups especially vulnerable to reduced service from pollinators. These results suggest that, without mitigating efforts, pollinator declines have the potential to reduce reproduction for most plant species, increasing the risk of population declines.

Seeing through the static: the temporal dimension of plant-animal mutualistic interactions.

Most studies of plant-animal mutualistic networks have come from a temporally static perspective. This approach has revealed general patterns in network structure, but limits our ability to understand the ecological and evolutionary processes that shape these networks and to predict the consequences of natural and human-driven disturbance on species interactions. We review the growing literature on temporal dynamics of plant-animal mutualistic networks including pollination, seed dispersal and ant defence mutualisms. We then discuss potential mechanisms underlying such variation in interactions, ranging from behavioural and physiological processes at the finest temporal scales to ecological and evolutionary processes at the broadest. We find that at the finest temporal scales (days, weeks, months) mutualistic interactions are highly dynamic, with considerable variation in network structure. At intermediate scales (years, decades), networks still exhibit high levels of temporal variation, but such variation appears to influence network properties only weakly. At the broadest temporal scales (many decades, centuries and beyond), continued shifts in interactions appear to reshape network structure, leading to dramatic community changes, including loss of species and function. Our review highlights the importance of considering the temporal dimension for understanding the ecology and evolution of complex webs of mutualistic interactions.

Uncovering patterns of freshwater positive interactions using meta-analysis: Identifying the roles of common participants, invasive species and environmental context.

Positive interactions are sensitive to human activities, necessitating synthetic approaches to elucidate broad patterns and predict future changes if these interactions are altered or lost. General understanding of freshwater positive interactions has been far outpaced by knowledge of these important relationships in terrestrial and marine ecosystems. We conducted a global meta-analysis to evaluate the magnitude of positive interactions across freshwater habitats. In 340 studies, we found substantial positive effects, with facilitators increasing beneficiaries by, on average, 81% across all taxa and response variables. Mollusks in particular were commonly studied as both facilitators and beneficiaries. Amphibians were one group benefiting the most from positive interactions, yet few studies investigated amphibians. Invasive facilitators had stronger positive effects on beneficiaries than non-invasive facilitators. We compared positive effects between high- and low-stress conditions and found no difference in the magnitude of benefit in the subset of studies that manipulated stressors. Future areas of research include understudied facilitators and beneficiaries, the stress gradient hypothesis, patterns across space or time and the influence of declining taxa whose elimination would jeopardise fragile positive interaction networks. Freshwater positive interactions occur among a wide range of taxa, influence populations, communities and ecosystem processes and deserve further exploration.

Longitudinal monitoring of honey bee colonies reveals dynamic nature of virus abundance and indicates a negative impact of Lake Sinai virus 2 on colony health.

Honey bees (Apis mellifera) are important pollinators of plants, including those that produce nut, fruit, and vegetable crops. Therefore, high annual losses of managed honey bee colonies in the United States and many other countries threaten global agriculture. Honey bee colony deaths have been associated with multiple abiotic and biotic factors, including pathogens, but the impact of virus infections on honey bee colony population size and survival are not well understood. To further investigate seasonal patterns of pathogen presence and abundance and the impact of viruses on honey bee colony health, commercially managed colonies involved in the 2016 California almond pollination event were monitored for one year. At each sample date, colony health and pathogen burden were assessed. Data from this 50-colony cohort study illustrate the dynamic nature of honey bee colony health and the temporal patterns of virus infection. Black queen cell virus, deformed wing virus, sacbrood virus, and the Lake Sinai viruses were the most readily detected viruses in honey bee samples obtained throughout the year. Analyses of virus prevalence and abundance revealed pathogen-specific trends including the overall increase in deformed wing virus abundance from summer to fall, while the levels of Lake Sinai virus 2 (LSV2) decreased over the same time period. Though virus prevalence and abundance varied in individual colonies, analyses of the overall trends reveal correlation with sample date. Total virus abundance increased from November 2015 (post-honey harvest) to the end of the almond pollination event in March 2016, which coincides with spring increase in colony population size. Peak total virus abundance occurred in late fall (August and October 2016), which correlated with the time period when the majority of colonies died. Honey bee colonies with larger populations harbored less LSV2 than weaker colonies with smaller populations, suggesting an inverse relationship between colony health and LSV2 abundance. Together, data from this and other longitudinal studies at the colony level are forming a better understanding of the impact of viruses on honey bee colony losses.

Land use and pollinator dependency drives global patterns of pollen limitation in the Anthropocene.

Land use change, by disrupting the co-evolved interactions between plants and their pollinators, could be causing plant reproduction to be limited by pollen supply. Using a phylogenetically controlled meta-analysis on over 2200 experimental studies and more than 1200 wild plants, we ask if land use intensification is causing plant reproduction to be pollen limited at global scales. Here we report that plants reliant on pollinators in urban settings are more pollen limited than similarly pollinator-reliant plants in other landscapes. Plants functionally specialized on bee pollinators are more pollen limited in natural than managed vegetation, but the reverse is true for plants pollinated exclusively by a non-bee functional group or those pollinated by multiple functional groups. Plants ecologically specialized on a single pollinator taxon were extremely pollen limited across land use types. These results suggest that while urbanization intensifies pollen limitation, ecologically and functionally specialized plants are at risk of pollen limitation across land use categories.

The indirect paths to cascading effects of extinctions in mutualistic networks.

Biodiversity loss is a hallmark of our times, but predicting its consequences is challenging. Ecological interactions form complex networks with multiple direct and indirect paths through which the impacts of an extinction may propagate. Here we show that accounting for these multiple paths connecting species is necessary to predict how extinctions affect the integrity of ecological networks. Using an approach initially developed for the study of information flow, we estimate indirect effects in plant-pollinator networks and find that even those species with several direct interactions may have much of their influence over others through long indirect paths. Next, we perform extinction simulations in those networks and show that although traditional connectivity metrics fail in the prediction of coextinction patterns, accounting for indirect interaction paths allows predicting species' vulnerability to the cascading effects of an extinction event. Embracing the structural complexity of ecological systems contributes towards a more predictive ecology, which is of paramount importance amid the current biodiversity crisis.

High-severity wildfire limits available floral pollen quality and bumble bee nutrition compared to mixed-severity burns.

High-severity wildfires, which can homogenize floral communities, are becoming more common relative to historic mixed-severity fire regimes in the Northern Rockies of the U.S. High-severity wildfire could negatively affect bumble bees, which are typically diet generalists, if floral species of inadequate pollen quality dominate the landscape post-burn. High-severity wildfires often require more time to return to pre-burn vegetation composition, and thus, effects of high-severity burns may persist past initial impacts. We investigated how wildfire severity (mixed- vs. high-severity) and time-since-burn affected available floral pollen quality, corbicular pollen quality, and bumble bee nutrition using percent nitrogen as a proxy for pollen quality and bumble bee nutrition. We found that community-weighted mean floral pollen nitrogen, corbicular pollen nitrogen, and bumble bee nitrogen were greater on average by 0.82%N, 0.60%N, and 1.16%N, respectively, in mixed-severity burns. This pattern of enhanced floral pollen nitrogen in mixed-severity burns was likely driven by the floral community, as community-weighted mean floral pollen percent nitrogen explained 87.4% of deviance in floral community composition. Only bee percent nitrogen varied with time-since-burn, increasing by 0.33%N per year. If these patterns persist across systems, our findings suggest that although wildfire is an essential ecosystem process, there are negative early successional impacts of high-severity wildfires on bumble bees and potentially on other pollen-dependent organisms via reductions in available pollen quality and nutrition. This work examines a previously unexplored pathway for how disturbances can influence native bee success via altering the nutritional landscape of pollen.

Nesting success of wood-cavity-nesting bees declines with increasing time since wildfire.

Bees require distinct foraging and nesting resources to occur in close proximity. However, spatial and temporal patterns in the availability and quantity of these resources can be affected by disturbances like wildfire. The potential for spatial or temporal separation of foraging and nesting resources is of particular concern for solitary wood-cavity-nesting bees as they are central-place, short-distance foragers once they have established their nest. Often the importance of nesting resources for bees have been tested by sampling foraging bees as a proxy, and nesting bees have rarely been studied in a community context, particularly postdisturbance. We tested how wood-cavity-nesting bee species richness, nesting success, and nesting and floral resources varied across gradients of wildfire severity and time-since-burn. We sampled nesting bees via nesting boxes within four wildfires in southwest Montana, USA, using a space-for-time substitution chronosequence approach spanning 3-25 years postburn and including an unburned control. We found that bee nesting success and species richness declined with increasing time postburn, with a complete lack of successful bee nesting in unburned areas. Nesting and floral resources were highly variable across both burn severity and time-since-burn, yet generally did not have strong effects on nesting success. Our results together suggest that burned areas may provide important habitat for wood-cavity-nesting bees in this system. Given ongoing fire regime shifts as well as other threats facing wild bee communities, this work helps provide essential information necessary for the management and conservation of wood-cavity-nesting bees.

Revealing biases in the sampling of ecological interaction networks.

The structure of ecological interactions is commonly understood through analyses of interaction networks. However, these analyses may be sensitive to sampling biases with respect to both the interactors (the nodes of the network) and interactions (the links between nodes), because the detectability of species and their interactions is highly heterogeneous. These ecological and statistical issues directly affect ecologists' abilities to accurately construct ecological networks. However, statistical biases introduced by sampling are difficult to quantify in the absence of full knowledge of the underlying ecological network's structure. To explore properties of large-scale ecological networks, we developed the software EcoNetGen, which constructs and samples networks with predetermined topologies. These networks may represent a wide variety of communities that vary in size and types of ecological interactions. We sampled these networks with different mathematical sampling designs that correspond to methods used in field observations. The observed networks generated by each sampling process were then analyzed with respect to the number of components, size of components and other network metrics. We show that the sampling effort needed to estimate underlying network properties depends strongly both on the sampling design and on the underlying network topology. In particular, networks with random or scale-free modules require more complete sampling to reveal their structure, compared to networks whose modules are nested or bipartite. Overall, modules with nested structure were the easiest to detect, regardless of the sampling design used. Sampling a network starting with any species that had a high degree (e.g., abundant generalist species) was consistently found to be the most accurate strategy to estimate network structure. Because high-degree species tend to be generalists, abundant in natural communities relative to specialists, and connected to each other, sampling by degree may therefore be common but unintentional in empirical sampling of networks. Conversely, sampling according to module (representing different interaction types or taxa) results in a rather complete view of certain modules, but fails to provide a complete picture of the underlying network. To reduce biases introduced by sampling methods, we recommend that these findings be incorporated into field design considerations for projects aiming to characterize large species interaction networks.