Principles for area-based biodiversity conservation.

Recent international agreements have strengthened and expanded commitments to protect and restore native habitats for biodiversity protection ("area-based biodiversity conservation"). Nevertheless, biodiversity conservation is hindered because how such commitments should be implemented has been strongly debated, which can lead to suboptimal habitat protection decisions. We argue that, despite the debates, there are three essential principles for area-based biodiversity conservation. These principles are related to habitat geographic coverage, amount, and connectivity. They emerge from evidence that, while large areas of nature are important and must be protected, conservation or restoration of multiple small habitat patches is also critical for global conservation, particularly in regions with high land use. We contend that the many area-based conservation initiatives expected in the coming decades should follow the principles we identify, regardless of ongoing debates. Considering the importance of biodiversity for maintenance of ecosystem services, we suggest that this would bring widespread societal benefits.

Automated acoustic detection of Geoffroy's spider monkey highlights tipping points of human disturbance.

As more land is altered by human activity and more species become at risk of extinction, it is essential that we understand the requirements for conserving threatened species across human-modified landscapes. Owing to their rarity and often sparse distributions, threatened species can be difficult to study and efficient methods to sample them across wide temporal and spatial scales have been lacking. Passive acoustic monitoring (PAM) is increasingly recognized as an efficient method for collecting data on vocal species; however, the development of automated species detectors required to analyse large amounts of acoustic data is not keeping pace. Here, we collected 35 805 h of acoustic data across 341 sites in a region over 1000 km2 to show that PAM, together with a newly developed automated detector, is able to successfully detect the endangered Geoffroy's spider monkey (Ateles geoffroyi), allowing us to show that Geoffroy's spider monkey was absent below a threshold of 80% forest cover and within 1 km of primary paved roads and occurred equally in old growth and secondary forests. We discuss how this methodology circumvents many of the existing issues in traditional sampling methods and can be highly successful in the study of vocally rare or threatened species. Our results provide tools and knowledge for setting targets and developing conservation strategies for the protection of Geoffroy's spider monkey.

The suggestion that landscapes should contain 40% of forest cover lacks evidence and is problematic.

A recent review suggests that forest cover needs to be restored or maintained on at least 40% of land area. In the absence of empirical evidence to support this threshold, we discuss how this suggestion is unhelpful and potentially dangerous. We advocate for regionally defined thresholds to inform conservation and restoration.

Support for the habitat amount hypothesis from a global synthesis of species density studies.

Decades of research suggest that species richness depends on spatial characteristics of habitat patches, especially their size and isolation. In contrast, the habitat amount hypothesis predicts that (1) species richness in plots of fixed size (species density) is more strongly and positively related to the amount of habitat around the plot than to patch size or isolation; (2) habitat amount better predicts species density than patch size and isolation combined, (3) there is no effect of habitat fragmentation per se on species density and (4) patch size and isolation effects do not become stronger with declining habitat amount. Data on eight taxonomic groups from 35 studies around the world support these predictions. Conserving species density requires minimising habitat loss, irrespective of the configuration of the patches in which that habitat is contained.

Edge effects on trophic cascades in tropical rainforests.

The cascading effects of biodiversity loss on ecosystem functioning of forests have become more apparent. However, how edge effects shape these processes has yet to be established. We assessed how edge effects alter arthropod populations and the strength of any resultant trophic cascades on herbivory rate in tropical forests of Brazil. We established 7 paired forest edge and interior sites. Each site had a vertebrate-exclosure, procedural (exclosure framework with open walls), and control plot (total 42 plots). Forest patches were surrounded by pasture. Understory arthropods and leaf damage were sampled every 4 weeks for 11 months. We used path analysis to determine the strength of trophic cascades in the interior and edge sites. In forest interior exclosures, abundance of predaceous and herbivorous arthropods increased by 326% and 180%, respectively, compared with control plots, and there were significant cascading effects on herbivory. Edge-dwelling invertebrates responded weakly to exclusion and there was no evidence of trophic cascade. Our results suggest that the vertebrate community at forest edges controls invertebrate densities to a lesser extent than it does in the interior. Edge areas can support vertebrate communities with a smaller contingent of insectivores. This allows arthropods to flourish and indirectly accounts for higher levels of plant damage at these sites. Increased herbivory rates may have important consequences for floristic community composition and primary productivity, as well as cascading effects on nutrient cycling. By interspersing natural forest patches with agroforests, instead of pasture, abiotic edge effects can be softened and prevented from penetrating deep into the forest. This would ensure a greater proportion of forest remains habitable for sensitive species and could help retain ecosystem functions in edge zones.

Efectos de Borde sobre las Cascadas Tróficas en los Bosques Tropicales Resumen Los efectos en cascada de la pérdida de la biodiversidad sobre el funcionamiento ecosistémico de los bosques se ha vuelto más evidente. A pesar de ésto, no se ha establecido cómo los efectos de borde moldean estos procesos. Evaluamos cómo los efectos de borde alteran las poblaciones de artrópodos y la fuerza de cualquier cascada trófica resultante sobre la tasa de herbivoría en los bosques tropicales de Brasil. Establecimos siete pares de sitios interiores y en el borde del bosque. Cada sitio tuvo un lote de encierro de vertebrados, uno procesal (un marco de trabajo de encierro con muros abiertos) y uno de control (total de 42 lotes). Los fragmentos de bosque estuvieron rodeados por potreros. Los artrópodos del sotobosque y el daño a las hojas fueron muestreados cada cuatro semanas durante once meses. Usamos el análisis de vía para determinar la fuerza de las cascadas tróficas en los sitios interiores y los del borde. En los encierros ubicados al interior del bosque la abundancia de artrópodos depredadores y herbívoros incrementó en un 326% y 180% respectivamente (comparada con los lotes de control) y hubo efectos relevantes de cascada sobre la herbivoría. Los invertebrados habitantes del borde respondieron débilmente al encierro y no hubo evidencia de la cascada trófica. Nuestros resultados sugieren que la comunidad de vertebrados en los bordes del bosque controla las densidades de invertebrados en un grado menor de lo que lo hace al interior del bosque. Las áreas del borde pueden mantener comunidades de vertebrados con un contingente menor de insectívoros. Ésto permite que los artrópodos florezcan y explica indirectamente los niveles más altos de daño a las plantas en estos sitios. El incremento de las tasas de herbivoría puede tener consecuencias importantes para la composición de la comunidad y para la productividad primaria, así como para los efectos cascada y el ciclo de nutrientes. Si intercalamos los fragmentos de bosque con agrobosques, en lugar de hacerlo con potreros, los efectos abióticos del borde pueden reducirse y se puede prevenir que penetren profundamente en el bosque. Esto aseguraría que una mayor proporción de bosque permanezca como habitable para las especies sensibles y podría ayudar a retener las funciones del ecosistema en las zonas de borde.

Mediation of area and edge effects in forest fragments by adjacent land use.

Habitat loss, fragmentation, and degradation have pervasive detrimental effects on tropical forest biodiversity, but the role of the surrounding land use (i.e., matrix) in determining the severity of these impacts remains poorly understood. We surveyed bird species across an interior-edge-matrix gradient to assess the effects of matrix type on biodiversity at 49 different sites with varying levels of landscape fragmentation in the Brazilian Atlantic Forest-a highly threatened biodiversity hotspot. Both area and edge effects were more pronounced in forest patches bordering pasture matrix, whereas patches bordering Eucalyptus plantation maintained compositionally similar bird communities between the edge and the interior and exhibited reduced effects of patch size. These results suggest the type of matrix in which forest fragments are situated can explain a substantial amount of the widely reported variability in biodiversity responses to forest loss and fragmentation.

Mediación de los Efectos de Área y de Borde sobre los Fragmentos de Bosque Causados por el Uso de Suelo Adyacente Resumen La pérdida del hábitat, la fragmentación y la degradación tienen efectos nocivos generalizados sobre la biodiversidad de los bosques tropicales. A pesar de esto, el papel del uso de suelo de los terrenos adyacentes (es decir, la matriz) en la determinación de la gravedad de estos impactos todavía está poco entendido. Censamos las especies de aves a lo largo de un gradiente de borde interno de matriz para evaluar los efectos del tipo de matriz sobre la biodiversidad en al menos 49 sitios con diferentes niveles de fragmentación del paisaje en el Bosque Atlántico Brasileño - un punto caliente de biodiversidad que se encuentra severamente amenazado. Tanto los efectos de área como los de borde estuvieron más pronunciados en los fragmentos de bosque que limitan con la matriz de pasturas, mientras que los fragmentos que limitan con plantaciones de Eucalyptus mantuvieron comunidades de aves similares en composición con aquellas entre el borde y el interior y mostraron efectos reducidos del tamaño de fragmento. Estos resultados sugieren que el tipo de matriz en el cual están situados los fragmentos de bosque puede explicar una cantidad sustancial de la ampliamente reportada variabilidad de respuestas a la pérdida del bosque y a la fragmentación.

Relationship between conservation biology and ecology shown through machine reading of 32,000 articles.

Conservation biology was founded on the idea that efforts to save nature depend on a scientific understanding of how it works. It sought to apply ecological principles to conservation problems. We investigated whether the relationship between these fields has changed over time through machine reading the full texts of 32,000 research articles published in 16 ecology and conservation biology journals. We examined changes in research topics in both fields and how the fields have evolved from 2000 to 2014. As conservation biology matured, its focus shifted from ecology to social and political aspects of conservation. The 2 fields diverged and now occupy distinct niches in modern science. We hypothesize this pattern resulted from increasing recognition that social, economic, and political factors are critical for successful conservation and possibly from rising skepticism about the relevance of contemporary ecological theory to practical conservation.

Relaciones entre la Biología de la Conservación y la Ecología Mostradas a través de la Lectura Mediante Máquina de 32,000 Artículos Resumen La biología de la conservación se fundó a partir de la idea de que los esfuerzos para salvar a la naturaleza dependen del entendimiento científico de cómo funciona. La biología de la conservación buscaba aplicar los principios ecológicos a los problemas de conservación. En este trabajo investigamos si la relación entre estos ámbitos ha cambiado con el tiempo al realizar una lectura mediante máquina de 32,000 textos completos de artículos de investigación publicados en 16 revistas sobre ecología y biología de la conservación. También examinamos los cambios en los temas de investigación en ambos ámbitos y cómo éstos han evolucionado desde el año 2000 hasta el 2014. Conforme ha madurado la biología de la conservación, su enfoque se ha movido de los aspectos ecológicos de la conservación a los aspectos políticos y sociales. La ecología y la biología de la conservación se han separado y ahora ocupan nichos distintos dentro de la ciencia moderna. Nuestra hipótesis considera que este patrón resultó de incrementar el reconocimiento de que los factores sociales, económicos y políticos son muy importantes para una conservación exitosa. Posiblemente el patrón también proviene del creciente escepticismo acerca de la relevancia que la teoría ecológica contemporánea tiene para la conservación en práctica.

ATLANTIC BIRDS: a data set of bird species from the Brazilian Atlantic Forest.

South America holds 30% of the world's avifauna, with the Atlantic Forest representing one of the richest regions of the Neotropics. Here we have compiled a data set on Brazilian Atlantic Forest bird occurrence (150,423) and abundance samples (N = 832 bird species; 33,119 bird individuals) using multiple methods, including qualitative surveys, mist nets, point counts, and line transects). We used four main sources of data: museum collections, on-line databases, literature sources, and unpublished reports. The data set comprises 4,122 localities and data from 1815 to 2017. Most studies were conducted in the Florestas de Interior (1,510 localities) and Serra do Mar (1,280 localities) biogeographic sub-regions. Considering the three main quantitative methods (mist net, point count, and line transect), we compiled abundance data for 745 species in 576 communities. In the data set, the most frequent species were Basileuterus culicivorus, Cyclaris gujanensis, and Conophaga lineata. There were 71 singletons, such as Lipaugus conditus and Calyptura cristata. We suggest that this small number of records reinforces the critical situation of these taxa in the Atlantic Forest. The information provided in this data set can be used for macroecological studies and to foster conservation strategies in this biodiversity hotspot. No copyright restrictions are associated with the data set. Please cite this Data Paper if data are used in publications and teaching events.

Trait-based indicators of bird species sensitivity to habitat loss are effective within but not across data sets.

Species' traits have been widely championed as the key to predicting which species are most threatened by habitat loss, yet previous work has failed to detect trends that are consistent enough to guide large-scale conservation and management. Here we explore whether traits and environmental variables predict species sensitivity to habitat loss across two data sets generated by independent avifaunal studies in the Atlantic Forest of Brazil, both of which detected a similar assemblage of species, and similar species-specific responses to habitat change, across an overlapping sample of sites. Specifically, we tested whether 25 distributional, climatic, ecological, behavioral, and morphological variables predict sensitivity to habitat loss among 196 bird species, both within and across studies, and when data were analysed as occurrence or abundance. We found that four to nine variables showed high explanatory power within a single study or data set, but none performed as strong predictors across all data sets. Our results demonstrate that the use of species traits to predict sensitivity to anthropogenic habitat loss can produce predictions that are species- and site-specific and not scalable to whole regions or biomes, and thus should be used with caution.

Atlantic forest bird communities provide different but not fewer functions after habitat loss.

Habitat loss often reduces the number of species as well as functional diversity. Dramatic effects to species composition have also been shown, but changes to functional composition have so far been poorly documented, partly owing to a lack of appropriate indices. We here develop three new community indices (i.e. functional integrity, community integrity of ecological groups and community specialization) to investigate how habitat loss affects the diversity and composition of functional traits and species. We used data from more than 5000 individuals of 137 bird species captured in 57 sites in the Brazilian Atlantic Forest, a highly endangered biodiversity hotspot.Results indicate that habitat loss leads to a decrease in functional integrity while measures of functional diversity remain unchanged or are even positively affected. Changes to functional integrity were caused by (i) a decrease in the provisioning of some functions, and an increase in others; (ii) strong within-guild species turnover; and (iii) a replacement of specialists by generalists. Hence, communities from more deforested sites seem to provide different but not fewer functions. We show the importance of investigating changes to both diversity and composition of functional traits and species, as the effects of habitat loss on ecosystem functioning may be more complex than previously thought. Crucially, when only functional diversity is assessed, important changes to ecological functions may remain undetected and negative effects of habitat loss underestimated, thereby imperiling the application of effective conservation actions.

Propagating variational model uncertainty for bioacoustic call label smoothing.

Along with propagating the input toward making a prediction, Bayesian neural networks also propagate uncertainty. This has the potential to guide the training process by rejecting predictions of low confidence, and recent variational Bayesian methods can do so without Monte Carlo sampling of weights. Here, we apply sample-free methods for wildlife call detection on recordings made via passive acoustic monitoring equipment in the animals' natural habitats. We further propose uncertainty-aware label smoothing, where the smoothing probability is dependent on sample-free predictive uncertainty, in order to downweigh data samples that should contribute less to the loss value. We introduce a bioacoustic dataset recorded in Malaysian Borneo, containing overlapping calls from 30 species. On that dataset, our proposed method achieves an absolute percentage improvement of around 1.5 points on area under the receiver operating characteristic (AU-ROC), 13 points in F1, and 19.5 points in expected calibration error (ECE) compared to the point-estimate network baseline averaged across all target classes.

Testing and optimizing metabarcoding of iDNA from dung beetles to sample mammals in the hyperdiverse Neotropics.

Over the past few years, insects have been used as samplers of vertebrate diversity by assessing the ingested-derived DNA (iDNA), and dung beetles have been shown to be a good mammal sampler given their broad feeding preference, wide distribution and easy sampling. Here, we tested and optimized the use of iDNA from dung beetles to assess the mammal community by evaluating if some biological and methodological aspects affect the use of dung beetles as mammal species samplers. We collected 403 dung beetles from 60 pitfall traps. iDNA from each dung beetle was sequenced by metabarcoding using two mini-barcodes (12SrRNA and 16SrRNA). We assessed whether dung beetles with different traits related to feeding, nesting and body size differed in the number of mammal species found in their iDNA. We also tested differences among four killing solutions in preserving the iDNA and compared the effectiveness of each mini barcode to recover mammals. We identified a total of 50 mammal OTUs (operational taxonomic unit), including terrestrial and arboreal species from 10 different orders. We found that at least one mammal-matching sequence was obtained from 70% of the dung beetle specimens. The number of mammal OTUs obtained did not vary with dung beetle traits as well as between the killing solutions. The 16SrRNA mini-barcode recovered a higher number of mammal OTUs than 12SrRNA, although both sets were partly non-overlapping. Thus, the complete mammal diversity may not be achieved by using only one of them. This study refines the methodology for routine assessment of tropical mammal communities via dung beetle 'samplers' and its universal applicability independently of the species traits of local beetle communities.

Climate-driven variation in dispersal ability predicts responses to forest fragmentation in birds.

Species sensitivity to forest fragmentation varies latitudinally, peaking in the tropics. A prominent explanation for this pattern is that historical landscape disturbance at higher latitudes has removed fragmentation-sensitive species or promoted the evolution of more resilient survivors. However, it is unclear whether this so-called extinction filter is the dominant driver of geographic variation in fragmentation sensitivity, particularly because climatic factors may also cause latitudinal gradients in dispersal ability, a key trait mediating sensitivity to habitat fragmentation. Here we combine field survey data with a morphological proxy for avian dispersal ability (hand-wing index) to assess responses to forest fragmentation in 1,034 bird species worldwide. We find that fragmentation sensitivity is strongly predicted by dispersal limitation and that other factors-latitude, body mass and historical disturbance events-have relatively limited explanatory power after accounting for species differences in dispersal. We also show that variation in dispersal ability is only weakly predicted by historical disturbance and more strongly associated with intra-annual temperature fluctuations (seasonality). Our results suggest that climatic factors play a dominant role in driving global variation in the impacts of forest fragmentation, emphasizing the need for more nuanced environmental policies that take into account local context and associated species traits.

Unraveling the drivers of community dissimilarity and species extinction in fragmented landscapes.

Communities in fragmented landscapes are often assumed to be structured by species extinction due to habitat loss, which has led to extensive use of the species-area relationship (SAR) in fragmentation studies. However, the use of the SAR presupposes that habitat loss leads species to extinction but does not allow for extinction to be offset by colonization of disturbed-habitat specialists. Moreover, the use of SAR assumes that species richness is a good proxy of community changes in fragmented landscapes. Here, we assessed how communities dwelling in fragmented landscapes are influenced by habitat loss at multiple scales; then we estimated the ability of models ruled by SAR and by species turnover in successfully predicting changes in community composition, and asked whether species richness is indeed an informative community metric. To address these issues, we used a data set consisting of 140 bird species sampled in 65 patches, from six landscapes with different proportions of forest cover in the Atlantic Forest of Brazil. We compared empirical patterns against simulations of over 8 million communities structured by different magnitudes of the power-law SAR and with species-specific rules to assign species to sites. Empirical results showed that, while bird community composition was strongly influenced by habitat loss at the patch and landscape scale, species richness remained largely unaffected. Modeling results revealed that the compositional changes observed in the Atlantic Forest bird metacommunity were only matched by models with either unrealistic magnitudes of the SAR or by models ruled by species turnover, akin to what would be observed along natural gradients. We show that, in the presence of such compositional turnover, species richness is poorly correlated with species extinction, and z values of the SAR strongly underestimate the effects of habitat loss. We suggest that the observed compositional changes are driven by each species reaching its individual extinction threshold: either a threshold of forest cover for species that disappear with habitat loss, or of matrix cover for species that benefit from habitat loss.

Associations of forest cover, fragment area, and connectivity with neotropical understory bird species richness and abundance.

Theoretical and empirical studies demonstrate that the total amount of forest and the size and connectivity of fragments have nonlinear effects on species survival. We tested how habitat amount and configuration affect understory bird species richness and abundance. We used mist nets (almost 34,000 net hours) to sample birds in 53 Atlantic Forest fragments in southeastern Brazil. Fragments were distributed among 3 10,800-ha landscapes. The remaining forest in these landscapes was below (10% forest cover), similar to (30%), and above (50%) the theoretical fragmentation threshold (approximately 30%) below which the effects of fragmentation should be intensified. Species-richness estimates were significantly higher (F= 3715, p = 0.00) where 50% of the forest remained, which suggests a species occurrence threshold of 30-50% forest, which is higher than usually occurs (<30%). Relations between forest cover and species richness differed depending on species sensitivity to forest conversion and fragmentation. For less sensitive species, species richness decreased as forest cover increased, whereas for highly sensitive species the opposite occurred. For sensitive species, species richness and the amount of forest cover were positively related, particularly when forest cover was 30-50%. Fragment size and connectivity were related to species richness and abundance in all landscapes, not just below the 30% threshold. Where 10% of the forest remained, fragment size was more related to species richness and abundance than connectivity. However, the relation between connectivity and species richness and abundance was stronger where 30% of the landscape was forested. Where 50% of the landscape was forested, fragment size and connectivity were both related to species richness and abundance. Our results demonstrated a rapid loss of species at relatively high levels of forest cover (30-50%). Highly sensitive species were 3-4 times more common above the 30-50% threshold than below it; however, our results do not support a unique fragmentation threshold.

The macroecology of landscape ecology.

One of landscape ecology's main goals is to unveil how biodiversity is impacted by habitat transformation. However, the discipline suffers from significant context dependency in observed spatial and temporal trends, hindering progress towards understanding the mechanisms driving species declines and preventing the development of accurate estimates of future biodiversity change. Here, we discuss recent evidence that populations' and species' responses to habitat change at the landscape scale are modulated by factors and processes occurring at macroecological scales, such as historical disturbance rates, distance to geographic range edges, and climatic suitability. We suggest that placing landscape ecology studies in a macroecological lens will help to explain seemingly inconsistent results and will ultimately create better predictive models to help mitigate the biodiversity crisis.

The magnitude and extent of edge effects on vascular epiphytes across the Brazilian Atlantic Forest.

Edge effects are ubiquitous landscape processes influencing over 70% of forest cover worldwide. However, little is known about how edge effects influence the vertical stratification of communities in forest fragments. We combined a spatially implicit and a spatially explicit approach to quantify the magnitude and extent of edge effects on canopy and understorey epiphytic plants in the Brazilian Atlantic Forest. Within the human-modified landscape, species richness, species abundance and community composition remained practically unchanged along the interior-edge gradient, pointing to severe biotic homogenisation at all strata. This is because the extent of edge effects reached at least 500 m, potentially leaving just 0.24% of the studied landscape unaffected by edges. We extrapolated our findings to the entire Atlantic Forest and found that just 19.4% of the total existing area is likely unaffected by edge effects and provide suitable habitat conditions for forest-dependent epiphytes. Our results suggest that the resources provided by the current forest cover might be insufficient to support the future of epiphyte communities. Preserving large continuous 'intact' forests is probably the only effective conservation strategy for vascular epiphytes.

Disentangling the effects of sampling scale and size on the shape of species abundance distributions.

Many authors have tried to explain the shape of the species abundance distribution (SAD). Some of them have suggested that sampling spatial scale is an important factor shaping SADs. These suggestions, however, did not consider the indirect and well-known effect of sample size, which increases as samples are combined to generate SADs at larger spatial scales. Here, we separate the effects of sample size and sampling scale on the shape of the SAD for three groups of organisms (trees, beetles and birds) sampled in the Brazilian Atlantic Forest. We compared the observed SADs at different sampling scales with simulated SADs having the same richness, relative abundances but comparable sample sizes, to show that the main effect shaping SADs is sample size and not sampling spatial scale. The effect of scale was minor and deviations between observed and simulated SADs were present only for beetles. For trees, the match between observed and simulated SADs was improved at all spatial scales when we accounted for conspecific aggregation, which was even more important than the sampling scale effect. We build on these results to propose a conceptual framework where observed SADs are shaped by three main factors, in decreasing order of importance: sample size, conspecific aggregation and beta diversity. Therefore, studies comparing SADs across sites or scales should use sampling and/or statistical approaches capable of disentangling these three effects on the shape of SADs.

How should we store avian faecal samples for microbiota analyses? Comparing efficacy and cost-effectiveness.

Analyses of bacterial DNA in faecal samples are becoming ever more common, yet we still do not know much about bird microbiomes. These challenges partly lie in the unique chemical nature of their faeces, and in the choice of sample storage method, which affects DNA preservation and the resulting microbiome composition. However, there is little information available on how best to preserve avian faeces for microbial analyses. This study evaluates five widely used methods for preserving nucleic acids and inferring microbiota profiles, for their relative efficacy, cost, and practicality. We tested the five methods (in-situ bead-beating with a TerraLyzer instrument, silica-bead desiccation, ethanol, refrigeration and RNAlater buffer) on 50 fresh faecal samples collected from captive House sparrows (Passer domesticus). In line with other studies, we find that different storage methods lead to distinct bacterial profiles. Storage method had a large effect on community composition and the relative abundance of dominant phyla such as Firmicutes and Proteobacteria, with the most significant changes observed for refrigerated samples. Furthermore, differences in the abundance of aerobic or facultatively aerobic taxa, particularly in refrigerated samples and those stored in ethanol, puts limits on comparisons of bacterial communities across different storage methods. Finally, the methods that did not include in-situ bead-beating did not recover comparable levels of microbiota to the samples that were immediately processed and preserved using a TerraLyzer device. However, this method is also less practical and more expensive under field work circumstances. Our study is the most comprehensive analysis to date on how storage conditions affect subsequent molecular assays applied to avian faeces and provides guidance on cost and practicality of methods under field conditions.