A maternal-effect Padi6 variant causes nuclear and cytoplasmic abnormalities in oocytes, as well as failure of epigenetic reprogramming and zygotic genome activation in embryos.

Maternal inactivation of genes encoding components of the subcortical maternal complex (SCMC) and its associated member, PADI6, generally results in early embryo lethality. In humans, SCMC gene variants were found in the healthy mothers of children affected by multilocus imprinting disturbances (MLID). However, how the SCMC controls the DNA methylation required to regulate imprinting remains poorly defined. We generated a mouse line carrying a Padi6 missense variant that was identified in a family with Beckwith-Wiedemann syndrome and MLID. If homozygous in female mice, this variant resulted in interruption of embryo development at the two-cell stage. Single-cell multiomic analyses demonstrated defective maturation of Padi6 mutant oocytes and incomplete DNA demethylation, down-regulation of zygotic genome activation (ZGA) genes, up-regulation of maternal decay genes, and developmental delay in two-cell embryos developing from Padi6 mutant oocytes but little effect on genomic imprinting. Western blotting and immunofluorescence analyses showed reduced levels of UHRF1 in oocytes and abnormal localization of DNMT1 and UHRF1 in both oocytes and zygotes. Treatment with 5-azacytidine reverted DNA hypermethylation but did not rescue the developmental arrest of mutant embryos. Taken together, this study demonstrates that PADI6 controls both nuclear and cytoplasmic oocyte processes that are necessary for preimplantation epigenetic reprogramming and ZGA.

A large net carbon loss attributed to anthropogenic and natural disturbances in the Amazon Arc of Deforestation.

The Amazon forest contains globally important carbon stocks, but in recent years, atmospheric measurements suggest that it has been releasing more carbon than it has absorbed because of deforestation and forest degradation. Accurately attributing the sources of carbon loss to forest degradation and natural disturbances remains a challenge because of the difficulty of classifying disturbances and simultaneously estimating carbon changes. We used a unique, randomized, repeated, very high-resolution airborne laser scanning survey to provide a direct, detailed, and high-resolution partitioning of aboveground carbon gains and losses in the Brazilian Arc of Deforestation. Our analysis revealed that disturbances directly attributed to human activity impacted 4.2% of the survey area while windthrows and other disturbances affected 2.7% and 14.7%, respectively. Extrapolating the lidar-based statistics to the study area (544,300 km2), we found that 24.1, 24.2, and 14.5 Tg C y-1 were lost through clearing, fires, and logging, respectively. The losses due to large windthrows (21.5 Tg C y-1) and other disturbances (50.3 Tg C y-1) were partially counterbalanced by forest growth (44.1 Tg C y-1). Our high-resolution estimates demonstrated a greater loss of carbon through forest degradation than through deforestation and a net loss of carbon of 90.5 ± 16.6 Tg C y-1 for the study region attributable to both anthropogenic and natural processes. This study highlights the role of forest degradation in the carbon balance for this critical region in the Earth system.

Macroscale controls determine the recovery of river ecosystem productivity following flood disturbances.

River ecosystem function depends on flow regimes that are increasingly modified by changes in climate, land use, water extraction, and flow regulation. Given the wide range of variation in flow regime modifications and autotrophic communities in rivers, it has been challenging to predict which rivers will be more resilient to flow disturbances. To better understand how river productivity is disturbed by and recovers from high-flow disturbance events, we used a continental-scale dataset of daily gross primary production time series from 143 rivers to estimate growth of autotrophic biomass and ecologically relevant flow disturbance thresholds using a modified population model. We compared biomass recovery rates across hydroclimatic gradients and catchment characteristics to evaluate macroscale controls on ecosystem recovery. Estimated biomass accrual (i.e., recovery) was fastest in wider rivers with less regulated flow regimes and more frequent instances of biomass removal during high flows. Although disturbance flow thresholds routinely fell below the estimated bankfull flood (i.e., the 2-y flood), a direct comparison of disturbance flows estimated by our biomass model and a geomorphic model revealed that biomass disturbance thresholds were usually greater than bed disturbance thresholds. We suggest that primary producers in rivers vary widely in their capacity to recover following flow disturbances, and multiple, interacting macroscale factors control productivity recovery rates, although river width had the strongest overall effect. Biomass disturbance flow thresholds varied as a function of geomorphology, highlighting the need for data such as bed slope and grain size to predict how river ecosystems will respond to changing flow regimes.

The maternal protein NLRP5 stabilizes UHRF1 in the cytoplasm: implication for the pathogenesis of multilocus imprinting disturbance.

We have recently discovered that the so-called subcortical maternal complex (SCMC) proteins composing of cytoplasmic lattices are destabilized in Uhrf1 knockout murine fully grown oocytes (FGOs). Here we report that human UHRF1 interacts with human NLRP5 and OOEP, which are core components of the SCMC. Moreover, NLRP5 and OOEP interact with DPPA3, which is an essential factor for exporting UHRF1 from the nucleus to the cytoplasm in oocytes. We identify that NLRP5, not OOEP, stabilizes UHRF1 protein in the cytoplasm utilizing specifically engineered cell lines mimicking UHRF1 status in oocytes and preimplantation embryos. Further, UHRF1 is destabilized both in the cytoplasm and nucleus of Nlrp5 knockout murine FGOs. Since pathogenic variants of the SCMC components frequently cause multilocus imprinting disturbance and UHRF1 is essential for maintaining CpG methylation of imprinting control regions during preimplantation development, our results suggest possible pathogenesis behind the disease, which has been a long-standing mystery.

Hurricane wind regimes for forests of North America.

Despite the ubiquity of tropical cyclones and their impacts on forests, little is known about how tropical cyclone regimes shape the ecology and evolution of tree species. We used a simple meteorological model (HURRECON) to estimate wind fields from hurricanes in the Western North Atlantic and Eastern North Pacific tropical cyclone basins from storms occurring between 1851 and 2022. We characterize how the intensity and frequency of hurricanes differ among geographically distinct hurricane regimes and define four hurricane regimes for North America (Continental, Inland, Coastal, and Fringe). Along this coastal-to-inland gradient, we found major differences in the frequency and intensity of hurricane wind regimes. The Fringe regime experiences category 1 winds relatively frequently [return period (RP) 25 y], whereas the Inland regime experiences category 1 winds very infrequently (RP ~3,000 y). We discuss how species traits related to tree windfirmness, such as mechanical properties and crown traits, may vary along hurricane regime gradients. Quantitative characterization of forest hurricane regimes provides a critical step for understanding the evolutionary and ecological role of hurricane regimes in wind-prone forests.

The hidden order of Turing patterns in arid and semi-arid vegetation ecosystems.

Vegetation Turing patterns play a critical role in the ecological functioning of arid and semi-arid ecosystems. However, the long-range spatial features of these patterns have been neglected compared to short-range features like patch shape and spatial wavelength. Drawing inspiration from hyperuniform structures in material science, we find that the arid and semi-arid vegetation Turing pattern exhibits long-range dispersion similar to hyperuniformity. As the degree of hyperuniformity of the vegetation Turing pattern increases, so does the water-use efficiency of the vegetation. This finding supports previous studies that suggest that Turing patterns represent a spatially optimized self-organization of ecosystems for water acquisition. The degree of hyperuniformity of Turing-type ecosystems exhibits significant critical slowing down near the tipping point, indicating that these ecosystems have non-negligible transient dynamical behavior. Reduced rainfall not only decreases the resilience of the steady state of the ecosystem but also slows down the rate of spatial optimization of water-use efficiency in long transient regimes. We propose that the degree of hyperuniformity indicates the spatial resilience of Turing-type ecosystems after strong, short-term disturbances. Spatially heterogeneous disturbances that reduce hyperuniformity lead to longer recovery times than spatially homogeneous disturbances that maintain hyperuniformity.

Warming-induced tree growth may help offset increasing disturbance across the Canadian boreal forest.

Large projected increases in forest disturbance pose a major threat to future wood fiber supply and carbon sequestration in the cold-limited, Canadian boreal forest ecosystem. Given the large sensitivity of tree growth to temperature, warming-induced increases in forest productivity have the potential to reduce these threats, but research efforts to date have yielded contradictory results attributed to limited data availability, methodological biases, and regional variability in forest dynamics. Here, we apply a machine learning algorithm to an unprecedented network of over 1 million tree growth records (1958 to 2018) from 20,089 permanent sample plots distributed across both Canada and the United States, spanning a 16.5 °C climatic gradient. Fitted models were then used to project the near-term (2050 s time period) growth of the six most abundant tree species in the Canadian boreal forest. Our results reveal a large, positive effect of increasing thermal energy on tree growth for most of the target species, leading to 20.5 to 22.7% projected gains in growth with climate change under RCP 4.5 and 8.5. The magnitude of these gains, which peak in the colder and wetter regions of the boreal forest, suggests that warming-induced growth increases should no longer be considered marginal but may in fact significantly offset some of the negative impacts of projected increases in drought and wildfire on wood supply and carbon sequestration and have major implications on ecological forecasts and the global economy.

High fried food consumption impacts anxiety and depression due to lipid metabolism disturbance and neuroinflammation.

Western dietary patterns have been unfavorably linked with mental health. However, the long-term effects of habitual fried food consumption on anxiety and depression and underlying mechanisms remain unclear. Our population-based study with 140,728 people revealed that frequent fried food consumption, especially fried potato consumption, is strongly associated with 12% and 7% higher risk of anxiety and depression, respectively. The associations were more pronounced among male and younger consumers. Consistently, long-term exposure to acrylamide, a representative food processing contaminant in fried products, exacerbates scototaxis and thigmotaxis, and further impairs exploration ability and sociality of adult zebrafish, showing anxiety- and depressive-like behaviors. Moreover, treatment with acrylamide significantly down-regulates the gene expression of tjp2a related to the permeability of blood-brain barrier. Multiomics analysis showed that chronic exposure to acrylamide induces cerebral lipid metabolism disturbance and neuroinflammation. PPAR signaling pathway mediates acrylamide-induced lipid metabolism disorder in the brain of zebrafish. Especially, chronic exposure to acrylamide dysregulates sphingolipid and phospholipid metabolism, which plays important roles in the development of anxiety and depression symptoms. In addition, acrylamide promotes lipid peroxidation and oxidation stress, which participate in cerebral neuroinflammation. Acrylamide dramatically increases the markers of lipid peroxidation, including (±)5-HETE, 11(S)-HETE, 5-oxoETE, and up-regulates the expression of proinflammatory lipid mediators such as (±)12-HETE and 14(S)-HDHA, indicating elevated cerebral inflammatory status after chronic exposure to acrylamide. Together, these results both epidemiologically and mechanistically provide strong evidence to unravel the mechanism of acrylamide-triggered anxiety and depression, and highlight the significance of reducing fried food consumption for mental health.

Circadian disturbances by altering the light-dark cycle negatively affects hematopoietic function of bone marrow in mice.

Circadian rhythms in metabolically active tissues are crucial for maintaining physical health. Circadian disturbance (CD) can cause various health issues, such as metabolic abnormalities and immune and cognitive dysfunctions. However, studies on the role of CD in immune cell development and differentiation, as well as the rhythmic expression of the core clock genes and their altered expression under CD, remain unclear. Therefore, we exposed C57bl/6j mice to repeated reversed light-dark cycles for 90 days to research the effects of CD on bone marrow (BM) hematopoietic function. We also researched the effects of CD on endogenous circadian rhythms, temporally dependent expression in peripheral blood and myeloid leukocytes, environmental homeostasis within BM, and circadian oscillations of hematopoietic-extrinsic cues. Our results confirmed that when the light and dark cycles around mice were frequently reversed, the circadian rhythmic expression of the two main circadian rhythm markers, the hypothalamic clock gene, and serum melatonin, was disturbed, indicating that the body was in a state of endogenous CD. Furthermore, CD altered the temporally dependent expression of peripheral blood and BM leukocytes and destroyed environmental homeostasis within the BM as well as circadian oscillations of hematopoietic-extrinsic cues, which may negatively affect BM hematopoiesis in mice. Collectively, these results demonstrate that circadian rhythms are vital for maintaining health and suggest that the association between CD and hematopoietic dysfunction warrants further investigation.

A neural mass model for disturbance of alpha rhythm in the minimal hepatic encephalopathy.

One of the early markers of minimal hepatic encephalopathy (MHE) is the disruption of alpha rhythm observed in electroencephalogram (EEG) signals. However, the underlying mechanisms responsible for this occurrence remain poorly understood. To address this gap, we develop a novel biophysical model MHE-AWD-NCM, encompassing the communication dynamics between a cortical neuron population (CNP) and an astrocyte population (AP), aimed at investigating the relationship between alpha wave disturbance (AWD) and mechanistical principles, specifically concerning astrocyte-neuronal communication in the context of MHE. In addition, we introduce the concepts of peak power density and peak frequency within the alpha band as quantitative measures of AWD. Our model faithfully reproduces the characteristic EEG phenomenology during MHE and shows how impairments of communication between CNP and AP could promote AWD. The results suggest that the disruptions in feedback neurotransmission from AP to CNP, along with the inhibition of GABA uptake by AP from the extracellular space, contribute to the observed AWD. Moreover, we found that the variation of external excitatory stimuli on CNP may play a key role in AWD in MHE. Finally, the sensitivity analysis is also performed to assess the relative significance of above factors in influencing AWD. Our findings align with the physiological observations and provide a more comprehensive understanding of the complex interplay of astrocyte-neuronal communication that underlies the AWD observed in MHE, which potentially may help to explore the targeted therapeutic interventions for the early stage of hepatic encephalopathy.

Behavioral responses to predatory sounds predict sensitivity of cetaceans to anthropogenic noise within a soundscape of fear.

SignificanceAcoustic signals travel efficiently in the marine environment, allowing soniferous predators and prey to eavesdrop on each other. Our results with four cetacean species indicate that they use acoustic information to assess predation risk and have evolved mechanisms to reduce predation risk by ceasing foraging. Species that more readily gave up foraging in response to predatory sounds of killer whales also decreased foraging more during 1- to 4-kHz sonar exposures, indicating that species exhibiting costly antipredator responses also have stronger behavioral reactions to anthropogenic noise. This advance in our understanding of the drivers of disturbance helps us to predict what species and habitats are likely to be most severely impacted by underwater noise pollution in oceans undergoing increasing anthropogenic activities.

Animal soundscapes reveal key markers of Amazon forest degradation from fire and logging.

Safeguarding tropical forest biodiversity requires solutions for monitoring ecosystem structure over time. In the Amazon, logging and fire reduce forest carbon stocks and alter habitat, but the long-term consequences for wildlife remain unclear, especially for lesser-known taxa. Here, we combined multiday acoustic surveys, airborne lidar, and satellite time series covering logged and burned forests (n = 39) in the southern Brazilian Amazon to identify acoustic markers of forest degradation. Our findings contradict expectations from the Acoustic Niche Hypothesis that animal communities in more degraded habitats occupy fewer "acoustic niches" defined by time and frequency. Instead, we found that aboveground biomass was not a consistent proxy for acoustic biodiversity due to the divergent patterns of "acoustic space occupancy" between logged and burned forests. Ecosystem soundscapes highlighted a stark, and sustained reorganization in acoustic community assembly after multiple fires; animal communication networks were quieter, more homogenous, and less acoustically integrated in forests burned multiple times than in logged or once-burned forests. These findings demonstrate strong biodiversity cobenefits from protecting burned Amazon forests from recurrent fire. By contrast, soundscape changes after logging were subtle and more consistent with acoustic community recovery than reassembly. In both logged and burned forests, insects were the dominant acoustic markers of degradation, particularly during midday and nighttime hours, which are not typically sampled by traditional biodiversity field surveys. The acoustic fingerprints of degradation history were conserved across replicate recording locations, indicating that soundscapes may offer a robust, taxonomically inclusive solution for digitally tracking changes in acoustic community composition over time.

Post-disturbance reorganization of forest ecosystems in a changing world.

Forest ecosystems are strongly impacted by continuing climate change and increasing disturbance activity, but how forest dynamics will respond remains highly uncertain. Here, we argue that a short time window after disturbance (i.e., a discrete event that disrupts prevailing ecosystem structure and composition and releases resources) is pivotal for future forest development. Trees that establish during this reorganization phase can shape forest structure and composition for centuries, providing operational early indications of forest change. While forest change has been fruitfully studied through a lens of resilience, profound ecological changes can be masked by a resilience versus regime shift dichotomy. We present a framework for characterizing the full spectrum of change after disturbance, analyzing forest reorganization along dimensions of forest structure (number, size, and spatial arrangement of trees) and composition (identity and diversity of tree species). We propose four major pathways through which forest cover can persist but reorganize following disturbance: resilience (no change in structure and composition), restructuring (structure changes but composition does not), reassembly (composition changes but structure does not), and replacement (structure and composition both change). Regime shifts occur when vegetation structure and composition are altered so profoundly that the emerging trajectory leads to nonforest. We identify fundamental processes underpinning forest reorganization which, if disrupted, deflect ecosystems away from resilience. To understand and predict forest reorganization, assessing these processes and the traits modulating them is crucial. A new wave of experiments, measurements, and models emphasizing the reorganization phase will further the capacity to anticipate future forest dynamics.

Increasing instability of a rocky intertidal meta-ecosystem.

Climate change threatens to destabilize ecological communities, potentially moving them from persistently occupied "basins of attraction" to different states. Increasing variation in key ecological processes can signal impending state shifts in ecosystems. In a rocky intertidal meta-ecosystem consisting of three distinct regions spread across 260 km of the Oregon coast, we show that annually cleared sites are characterized by communities that exhibit signs of increasing destabilization (loss of resilience) over the past decade despite persistent community states. In all cases, recovery rates slowed and became more variable over time. The conditions underlying these shifts appear to be external to the system, with thermal disruptions (e.g., marine heat waves, El Niño-Southern Oscillation) and shifts in ocean currents (e.g., upwelling) being the likely proximate drivers. Although this iconic ecosystem has long appeared resistant to stress, the evidence suggests that subtle destabilization has occurred over at least the last decade.

Bace1 Deletion in the Adult Reverses Epileptiform Activity and Sleep-wake Disturbances in AD Mice.

Alzheimer's disease (AD) increases the risk for seizures and sleep disorders. We show here that germline deletion of ß-site amyloid precursor protein (APP) cleaving enzyme-1 (BACE1) in neurons, but not in astrocytes, increased epileptiform activity. However, Bace1 deletion at adult ages did not alter the normal EEG waveform, indicating less concern for BACE1 inhibition in patients. Moreover, we showed that deletion of Bace1 in the adult was able to reverse epileptiform activity in 5xFAD mice. Intriguingly, treating 5xFAD and APPNL-G-F/NL-G-F (APP KI) mice of either sex with one BACE1 inhibitor Lanabecestat (AZD3293) dramatically increased epileptiform spiking, likely resulting from an off-target effect. We also monitored sleep-wake pathologies in these mice and showed increased wakefulness, decreased non-rapid eye movement sleep, and rapid eye movement sleep in both 5xFAD and APP KI mice; BACE1 inhibition in the adult 5xFAD mice reversed plaque load and sleep disturbances, but this was not seen in APP KI mice. Further studies with and without BACE1 inhibitor treatment showed different levels of plaque-associated microgliosis and activated microglial proteins in 5xFAD mice compared with APP KI mice. Together, BACE1 inhibition should be developed to avoid off-target effect for achieving benefits in reducing epileptic activity and sleep disturbance in Alzheimer's patients.SIGNIFICANCE STATEMENT BACE1 is widely recognized as a therapeutic target for treating Alzheimer's disease patients. However, BACE1 inhibitors failed in clinical trials because of inability to show cognitive improvement in patients. Here we show that BACE1 inhibition actually reduces sleep disturbances and epileptic seizures; both are seen in AD patients. We further showed that one of clinically tested BACE1 inhibitors does have off-target effects, and development of safer BACE1 inhibitors will be beneficial to AD patients. Results from this study will provide useful guidance for additional drug development.

Frequency-dependent tolerance to aircraft disturbance drastically alters predicted impact on shorebirds.

Anthropogenic disturbance of wildlife is increasing globally. Generalizing impacts of disturbance to novel situations is challenging, as the tolerance of animals to human activities varies with disturbance frequency (e.g. due to habituation). Few studies have quantified frequency-dependent tolerance, let alone determined how it affects predictions of disturbance impacts when these are extrapolated over large areas. In a comparative study across a gradient of air traffic intensities, we show that birds nearly always fled (80%) if aircraft were rare, while birds rarely responded (7%) if traffic was frequent. When extrapolating site-specific responses to an entire region, accounting for frequency-dependent tolerance dramatically alters the predicted costs of disturbance: the disturbance map homogenizes with fewer hotspots. Quantifying frequency-dependent tolerance has proven challenging, but we propose that (i) ignoring it causes extrapolations of disturbance impacts from single sites to be unreliable, and (ii) it can reconcile published idiosyncratic species- or source-specific disturbance responses.

Disturbances can facilitate prior invasions more than subsequent invasions in microbial communities.

Invasions are commonly found to benefit from disturbance events. However, the importance of the relative timing of the invasion and disturbance for invader success and impact on community composition remains uncertain. Here, we experimentally test this by invading a five-species bacterial community on eight separate occasions-four before a disturbance and four after. Invader success and impact on community composition was greatest when the invasion immediately followed the disturbance. However, the subsequent invasions had negligible success or impact. Pre-disturbance, invader success and impact was greatest when the invader was added just before the disturbance. Importantly, however, the first three pre-disturbance invasion events had significantly greater success than the last three post-disturbance invasions. Moreover, these findings were consistent across a range of propagule pressures. Overall, we demonstrate that timing is highly important for both the success and impact on community composition of an invader, with both being lower as time since disturbance progresses.

Arrive and wait: Inactive bacterial taxa contribute to perceived soil microbiome resilience after a multidecadal press disturbance.

Long-term (press) disturbances like the climate crisis and other anthropogenic pressures are fundamentally altering ecosystems and their functions. Many critical ecosystem functions, such as biogeochemical cycling, are facilitated by microbial communities. Understanding the functional consequences of microbiome responses to press disturbances requires ongoing observations of the active populations that contribute to functions. This study leverages a 7-year time series of a 60-year-old coal seam fire (Centralia, Pennsylvania, USA) to examine the resilience of soil bacterial microbiomes to a press disturbance. Using 16S rRNA and 16S rRNA gene amplicon sequencing, we assessed the interannual dynamics of the active subset and the 'whole' bacterial community. Contrary to our hypothesis, the whole communities demonstrated greater resilience than active subsets, suggesting that inactive members contributed to overall structural resilience. Thus, in addition to selection mechanisms of active populations, perceived microbiome resilience is also supported by mechanisms of dispersal, persistence, and revival from the local dormant pool.

Long-term grazing changed the spatial distributions of dominant species in typical steppe of Inner Mongolia.

Dominant species occupy a pivotal role in plant community, influencing the structure and function of the ecosystem. The spatial distributions of dominant species can react to the effect of different grazing intensities, thereby reflecting their tolerance and adaptive strategies toward grazing. In this study, geostatistical methods were mainly used to study the spatial distribution characteristics of Stipa krylovii Roshev. and Leymus chinensis (Trin.) Tzvel. species at two interval scales (quadrat size 5 m × 5 m, 10 m × 10 m) and two treatments (free grazing, FG, 1.66 sheep·ha- 1·a- 1; control, CK, 0 sheep·ha- 1·a- 1) in typical steppe of Inner Mongolia. A systematic sampling method was used in each 100 m × 100 m representative sample plots to obtain the height, coverage, and density of all species in the community. The results showed that grazing altered the concentrated distribution of S. krylovii and the spatial mosaic distribution pattern of S. krylovii and L. chinensis while having no effect on the spatial clumped distribution of L. chinensis. It also found that the spatial distributions of dominant species are primarily affected by structural factors, and random factors such as long-term grazing led to a transition of S. krylovii from a concentrated distribution to a small patchy random pattern should not be overlooked. Our findings suggest that long-term grazing alters the spatial distribution pattern of dominant species and that adaptive strategies may be the key for maintaining the dominant role of structural factors.

Body mass mediates spatio-temporal responses of mammals to human frequentation across Italian protected areas.

Protected area (PA) networks are a pivotal tool to fight biodiversity loss, yet they often need to balance the mission of nature conservation with the socio-economic need of giving opportunity for outdoor recreation. Recreation in natural areas is important for human health in an urbanized society, but can prompt behavioural modifications in wild animals. Rarely, however, have these responses being studied across multiple PAs and using standardized methods. We deployed a systematic camera trapping protocol at over 200 sites to sample medium and large mammals in four PAs within the European Natura 2000 network to assess their spatio-temporal responses to human frequentation, proximity to towns, amount of open habitat and topographical variables. By applying multi-species and single-species models for the number of diurnal, crepuscular and nocturnal detections and a multi-species model for nocturnality index, we estimated both species-specific- and meta-community-level effects, finding that increased nocturnality appeared the main strategy that the mammal meta-community used to cope with human disturbance. However, responses in the diurnal, crepuscular and nocturnal site use were mediated by species' body mass, with larger species exhibiting avoidance of humans and smaller species more opportunistic behaviours. Our results show the effectiveness of standardized sampling and provide insights for planning the expansion of PA networks as foreseen by the Kunming-Montreal biodiversity agreement.