Biology subjects

Ferreira, A. C.

Publications and source records attributed to Ferreira, A. C..

17 recordsLinked to original sources

bioRxiv2025

Infanticide in a colonial cooperative bird is not associated with direct reproductive benefits

Infanticide, the killing of dependent young by adult conspecifics, is an extreme form of conflict that remains poorly understood. Previous studies suggest that it is associated with reproductive benefits, either by making breeders newly available for reproduction or by increasing offspring care and reducing competition for resources. In addition, infanticide may result from a reproductive trade-off, where parents kill their offspring if environmental conditions compromise their reproductive value. Here, we combine video, genetic and individual-level reproductive data collected over 12 years to describe infanticide in a highly social, cooperative bird, the sociable weaver Philetairus socius and to establish whether direct reproductive benefits or life-history trade-offs are possible motivators for this behaviour. We documented 50 cases of infanticide and identified 27 aggressors with known histories. These were mostly born outside the colonies where they committed infanticide (63%) and there was a bias towards younger individuals. Aggressors were usually not part of the victims breeding group (74%), but in two cases infanticide was committed by the breeding female, and in five by helpers. In most cases, aggressors were not reproducing at the time of the infanticide, were not recorded mating with any of the breeders or helpers from the attacked nest, nor occupied the nest from where the chicks were evicted. Only 13 of the known aggressors were confirmed as breeders after the attacks, and these mostly reproduced at the colony where the attacks were committed. Overall, we found no evidence for direct reproductive benefits of infanticide in sociable weavers. However, the younger age of aggressors, lack of genetic relatedness to the victims and future breeding at the colony suggest that competition between breeding groups, and hence indirect reproductive benefits, could provide motivation for infanticide in this population. Additional work over longer periods is needed to identify these possible indirect fitness benefits of infanticide.

animal behavior and cognition↗

bioRxiv2025

Youth-associated protein TIMP2 alters microglial state and function in the context of aging

There is little understanding of how aging serves as the strongest risk factor for several neurogenerative diseases. Specific neural cell types, such as microglia, undergo age-related maladaptive changes, including increased inflammation, impaired debris clearance, and cellular senescence, yet specific mediators that regulate these processes remain unclear. The aged brain is rejuvenated by youth-associated plasma factors, including tissue inhibitor of metalloproteinases 2 (TIMP2), which we have shown acts on the extracellular matrix (ECM) to regulate synaptic plasticity. Given emerging roles for microglia in these processes, we examined the impact of TIMP2 on microglial function. We show that TIMP2 deletion exacerbates microglial phenotypes associated with aging, including transcriptomic changes in cell activation, increased microgliosis, and increased levels of stress and inflammatory proteins measured in the brain extracellular space by in vivo microdialysis. Deleting specific cellular pools of TIMP2 in vivo increased microglial activation and altered myelin phagocytosis. Treating aged mice with TIMP2 reversed several phenotypes observed in our deletion models, resulting in decreased microglial activation, reduced proportions of proinflammatory microglia, and enhanced phagocytosis of physiological substrates. Our results identify TIMP2 as a key modulator of age-associated microglia dysfunction. Harnessing its activity may mitigate detrimental effects of age-associated insults on microglia function.

neuroscience↗

bioRxiv2025

Long-term evaluation of the association between dominance, bib size, sex and age

The association between dominance and phenotypic traits ( badges of status) has been recently questioned, since it has been found to show more variation across populations and time than expected. Using an eleven-year dataset, encompassing more than 1,800 individuals, we studied the association between the size of a melanin-based plumage trait (a black patch under the beak or bib), individual attributes (age and sex) and dominance. Our study model, the sociable weaver (Philetairus socius), is a small passerine, which lives year-round in colonies of varying size. Bib sizes were obtained from pictures and automatically pre-processed with an object segmentation deep learning network. We recorded 36,346 dominance-related interactions to estimate dominance hierarchies. We then investigated the effect of sex and age on bib size, and the overall and between-year association of bib size and age with dominance for each sex. We found that bib size increased with age within individuals of both sexes. Age was a strong and consistent predictor of dominance in males, but not in females. Nevertheless, dominance was not correlated with bib size in males and in females. Finally, these relationships did not strongly vary between years. Our results suggest that males and females may use different factors to regulate dominance interactions. Furthermore, our longitudinal dataset allowed us to show that bib size is not a badge of status in our population even though its growth encodes information on age. These results highlight the importance of considering multiple traits over time when studying morphological signalling of dominance hierarchy.

animal behavior and cognition↗

bioRxiv2024

Foraging associations are related with helping interactions in a cooperatively breeding bird

Kin selection has been the main hypothesis explaining helping behaviour in cooperative breeders, with evidence being largely based on the observation that helpers tend to provide to related offspring. However, kin-biased help could conceal additional, mechanisms contributing to the maintenance of cooperation. Under pay-to-stay, group augmentation and partner choice hypotheses, a range of direct benefits can arise through helping. Here, we explored this potential mechanism by testing whether the social associations of breeding individuals were related with the help that they received from non-breeding individuals. We collected social associations from PIT-tagged sociable weavers, Philetairus socius, at RFID feeding stations, which allowed us to compare associations between breeders and either their helpers (mostly kin) or their other kin that did not help--before, during and after reproduction. Using correlative tests and data-driven simulations, we show that helpers have stronger foraging bonds with breeders than non-helping kin, and that these stronger bonds are present both prior and post breeding. Furthermore, helper-breeding female social affiliations were positively correlated with the amount of help provided. Our results suggest that direct benefits of social associations complement kin selection to determine helping decision, and that these in turn influence future social associations.

animal behavior and cognition↗

bioRxiv2024

From raw footage to behaviour: an automated pipeline for activity monitoring and behaviour identification in the wild

O_LIStudies of animal behaviour usually rely on direct observations or manual annotations of video recordings. However, such methods can be very time-consuming and error-prone, leading to sub-optimal sample sizes. Recent advances in deep-learning show great potential to overcome such limitations, nevertheless, most currently available behavioural recognition solutions remain focused on captivity settings. C_LIO_LIHere, we present a deployment-focused framework to guide researchers in building behavioural recognition systems from video data, using Long Short-Term Memory (LSTM) networks to classify behavioural sequences across consecutive frames. C_LIO_LILSTMs allowed to: 1) monitor nest activity by detecting the birds presence and simultaneously classifying the type of trajectory: i.e., nest-chamber entrance or exit; and 2) identify the behaviour performed: building, aggression or sanitation. Using our framework, we outperformed human annotators when jointly considering error and speed. Model performance improved with challenging training instances, and remained robust even with modest sample sizes. LSTM also outperformed YOLO ("You Only Look Once"), highlighting the critical role of temporal sequence information in behavioural analysis. C_LIO_LIWe demonstrate that our approach is replicable across three bird species and applicable to deployment videos, highlighting its value as a generalizable and transferable tool for long-term studies in the wild. C_LI DATA AVAILABILITYScripts, models, and data required to reproduce this work are available on Zenodo (DOIs: 10.5281/zenodo.18681623 and 10.5281/zenodo.18695178).

animal behavior and cognition↗

bioRxiv2024

A deep learning approach to detect and visualise sexual dimorphism in monomorphic species

Sex recognition is facilitated by dimorphism in some traits. However, humans often fail to find the traits that allow to distinguish between sexes in other species. Deep learning has the potential to surpass humans in identifying cryptic differences between sexes, but, so far, has rarely been used to assess sexual dimorphism. In this study, we evaluated (i) the ability of a fine-tuned classification neural network, EfficientNet, to find differences between sexes in a species that appears monomorphic to humans, the sociable weaver (Philetairus socius). We then assessed (ii) the benefits of Grad-CAM visualisation techniques to understand which parts of the individuals are used by the network to differentiate the sexes. We trained 10-folds cross-validation models on more than 4,500 pictures of the head from more than 1,300 individuals. Our results show that the network can predict sex of sociable weavers with an accuracy of 76%, which is considerably higher than humans performance, and that the model was similarly good at predicting females and males. When interpreting the probability of being classified to one sex, our results further reveal an effect of the interaction of sex with age on the confidence score of the models which shows that younger males are less masculine than older ones, and older females more masculine than younger ones. Finally, using Grad-CAM we found that the model mostly uses the beak region to predict the sex of individuals. Overall, this work shows that artificial intelligence has the potential to be a non-invasive sexing tool, surpassing human capabilities and aiding in pinpointing potential cryptic dimorphic body parts that have yet to be identified. In birds, half of the worlds species appear sexually monomorphic to humans, and re-evaluation of species dimorphism with this type of methods could deepen our understanding of the effect of selection on animal traits.

zoology↗

bioRxiv2022

Neuronal TIMP2 regulates hippocampus-dependent plasticity and extracellular matrix complexity

The functional output of the hippocampus, a brain region subserving memory processes, depends on highly orchestrated cellular and molecular processes that regulate synaptic plasticity throughout life. The structural requirements of such plasticity and molecular processes involved in this regulation are poorly understood. Specific molecules, including tissue inhibitor of metalloproteinases-2 (TIMP2) have been implicated in processes of plasticity in the hippocampus, a role that decreases with brain aging as expression is lost. Here, we report that TIMP2 is highly expressed by neurons within the hippocampus and its loss drives changes in cellular programs related to adult neurogenesis and dendritic spine turnover with corresponding impairments in hippocampus-dependent memory. Consistent with the accumulation of ECM in the hippocampus we observe with aging, we find that TIMP2 acts to reduce accumulation of extracellular matrix (ECM) around synapses in the hippocampus. Moreover, its removal results in hindrance of newborn neuron migration through a denser ECM network. A novel conditional TIMP2 KO mouse reveals that neuronal TIMP2 regulates adult neurogenesis, accumulation of ECM, and ultimately hippocampus-dependent memory. Our results define a mechanism whereby hippocampus- dependent function is regulated by TIMP2 and its interactions with the ECM to regulate diverse processes associated with synaptic plasticity.

neuroscience↗

bioRxiv2022

Macrophages undergo necroptosis during severe influenza A infection and contribute to virus-associated cytokine storm

Influenza A virus (IAV) causes a major public health concern, because it is one of the leading causes of respiratory tract infections and hospitalization. Severe influenza has been associated with the cytokine storm, and IAV productive infection cell death in airway epithelial cells may contribute to the exacerbation of this proinflammatory event. On the other hand, IAV replication in macrophages is non-permissive and whether this immune cell may contribute to severe influenza physiopathology requires more details. Here, we investigated IAV-induced macrophage death, along with potential therapeutic intervention. We found that IAV or simply its surface glycoprotein hemagglutinin (HA) triggers necroptosis in human and murine macrophages in a Toll-like receptor-4 (TLR4) and TNF-dependent manner. Anti-TNF treatment, with the clinically approved drug etanercept, prevented the engagement of the necroptotic loop and mice mortality. impaired IAV-induced pro-inflammatory cytokine storm and lung injury. Our results implicate macrophage necroptosis with severe influenza in experimental models and potentially repurpose a clinically available therapy. Author SummaryVarious fates of cell death have an integral role in the influenza A virus (IAV) pathogenesis and lung/respiratory dysfunction. IAV physiopathology is not restricted to airway epithelial cells, where this virus actively replicated. Macrophages should support both viral clearance and priming of adaptative immune response in patients that adequately control influenza. However, during severe IAV infection, macrophages - which are unable to support a permissive viral replication - undergo disruptive cell death and contribute to the exacerbated production of proinflammatory cytokines/chemokines. We characterized this process by showing that IAV or just its surface glycoprotein hemagglutinin (HA) trigger necroptosis, a disruptive and TNF-dependent cells death. Since TNF is a hallmark of pro-inflammatory cell death, we blocked this mediator with a repurposed biomedicine etanercept, which prevented the severe IAV infection in the experimental model. The present work improves the knowledge of influenza pathophysiology by highlighting the importance of macrophage cell death during severe infection.

immunology↗

bioRxiv2021

Atazanavir is a competitive inhibitor of SARS-CoV-2 Mpro, impairing variants replication in vitro and in vivo

Atazanavir (ATV) has already been considered as a potential repurposing drug to 2019 coronavirus disease (COVID-19), however, there are controversial reports on its mechanism of action and effectiveness as anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Through the pre-clinical chain of experiments: enzymatic, molecular docking, cell-based, and in vivo assays, it is demonstrated here that both SARS-CoV-2 B.1 lineage and variant of concern gamma are susceptible to this antiretroviral. Enzymatic assays and molecular docking calculations showed that SARS-CoV-2 main protease (Mpro) was inhibited by ATV, with Morrisons inhibitory constant (Ki) 1.5-fold higher than boceprevir (GC376, a positive control). ATV was a competitive inhibition, increasing the Mpros Michaelis-Menten (Km) more than 6-fold. Cell-based assays indicated that SARS-CoV-2 gamma is more susceptible to ATV than its predecessor strain B.1. Using oral administration of ATV in mice to reach plasmatic exposure similar to humans, transgenic mice expression in human angiotensin converting enzyme 2 (K18-hACE2) were partially protected against lethal challenge with SARS-CoV-2 gamma. Moreover, less cell death and inflammation were observed in the lung from infected and treated mice. Our studies may contribute to a better comprehension of the Mpro/ATV interaction, which could pave the way to the development of specific inhibitors of this viral protease.

pharmacology and toxicology↗

bioRxiv2021

Unlike Chloroquine, mefloquine inhibits SARS-CoV-2 infection in physiologically relevant cells and does not induce viral variants.

Repositioning of clinical approved drugs could represent the fastest way to identify therapeutic options during public health emergencies, the majority of drugs explored for repurposing as antivirals for 2019 coronavirus disease (COVID-19) have failed to demonstrate clinical benefit. Without specific antivirals, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic continues to cause major global mortality. Antimalarial drugs, such as chloroquine (CQ)/hydroxychloroquine (HCQ) and mefloquine have emerged as potential anti-SARS-CoV-2 antivirals. CQ/HCQ entered the Solidarity and RECOVERY clinical trials against COVID-19 and showed lack of efficacy. Importantly, mefloquine is not a 4-aminoquinoline like CQ and HCQ and has been previously repurposed for other respiratory diseases. Unlike the 4-aminoquinolines that accumulate in the high pH of intracellular lysosomes of the lung, the high respiratory tract penetration of mefloquine is driven by its high lipophilicity. While CQ and HCQ exhibit activity in Vero E6 cells, their activity is obviated in TMPRSS2-expressing cells, such as Calu-3 cells, which more accurately recapitulate in vivo entry mechanisms for SARS-CoV-2. Accordingly, here we report the anti-SARS-CoV-2 activity of mefloquine in Calu-3 type II pneumocytes and primary human monocytes. Mefloquine inhibited SARS-CoV-2 replication in Calu-3 cells with low cytotoxicity and EC50 and EC90 values of 1.2 and 5.3 {micro}M, respectively. In addition, mefloquine reduced up to 68% the SARS-CoV-2 RNA levels in infected monocytes, reducing viral-induced inflammation. Mefloquine blocked early steps of the SARS-CoV-2 replicative cycle and was less prone than CQ to induce drug-associated viral mutations and synergized with RNA polymerase inhibitor. The pharmacological parameters of mefloquine are consistent with its plasma exposure in humans and its tissue-to-plasma predicted coefficient points that this drug may accumulate in the lungs. These data indicate that mefloquine could represent an orally available clinically approved drug option against COVID-19 and should not be neglected on the basis of the failure of CQ and HCQ.

microbiology↗

bioRxiv2021

A suitable murine model for studying respiratory coronavirus infection and therapeutic countermeasures in BSL-2 laboratories

Several animal models are being used to explore important features of COVID-19, nevertheless none of them recapitulates all aspects of the disease in humans. The continuous refinement and development of other options of in vivo models are opportune, especially ones that are carried out at BSL-2 (Biosafety Level 2) laboratories. In this study, we investigated the suitability of the intranasal infection with the murine betacoronavirus MHV-3 to recapitulate multiple aspects of the pathogenesis of COVID-19 in C57BL/6J mice. We demonstrate that MHV-3 replicated in lungs 1 day after inoculation and triggered respiratory inflammation and dysfunction. This MHV-model of infection was further applied to highlight the critical role of TNF in cytokine-mediated coronavirus pathogenesis. Blocking TNF signaling by pharmacological and genetic strategies greatly increased the survival time and reduces lung injury of MHV-3-infected mice. In vitro studies showed that TNF blockage decreased SARS-CoV-2 replication in human epithelial lung cells and resulted in the lower release of IL-6 and IL-8 cytokines beyond TNF itself. Taken together, our results demonstrate that this model of MHV infection in mice is a useful BSL-2 screening platform for evaluating pathogenesis for human coronaviruses infections, such as COVID-19.

microbiology↗

bioRxiv2020

Lipid droplets fuels SARS-CoV-2 replication and inflammatory response

Viruses are obligate intracellular parasites that make use of the host metabolic machineries to meet their biosynthetic needs, identifying the host pathways essential for the virus replication may lead to potential targets for therapeutic intervention. The mechanisms and pathways explored by SARS-CoV-2 to support its replication within host cells are not fully known. Lipid droplets (LD) are organelles with major functions in lipid metabolism and energy homeostasis, and have multiple roles in infections and inflammation. Here we described that monocytes from COVID-19 patients have an increased LD accumulation compared to SARS-CoV-2 negative donors. In vitro, SARS-CoV-2 infection modulates pathways of lipid synthesis and uptake, including CD36, SREBP-1, PPAR{gamma} and DGAT-1 in monocytes and triggered LD formation in different human cells. LDs were found in close apposition with SARS-CoV-2 proteins and double-stranded (ds)-RNA in infected cells. Pharmacological modulation of LD formation by inhibition of DGAT-1 with A922500 significantly inhibited SARS-CoV-2 replication as well as reduced production of pro-inflammatory mediators. Taken together, we demonstrate the essential role of lipid metabolic reprograming and LD formation in SARS-CoV-2 replication and pathogenesis, opening new opportunities for therapeutic strategies to COVID-19.

immunology↗

bioRxiv2020

The neuropeptides VIP and PACAP inhibit SARS-CoV-2 replication in monocytes and lung epithelial cells and decrease the production of proinflammatory cytokines in infected cells.

Infection by SARS-CoV-2 may elicit uncontrolled and damaging inflammatory responses. Thus, it is critical to identify compounds able to inhibit virus replication and thwart the inflammatory reaction. Here, we show that the plasma levels of the immunoregulatory neuropeptide VIP are elevated in patients with severe COVID-19, correlating with reduced inflammatory mediators and with survival on those patients. In vitro, VIP and PACAP, highly similar neuropeptides, decreased the SARS-CoV-2 genome replication in human monocytes and viral production in lung epithelial cells, also reducing cell death. Both neuropeptides inhibited the production of proinflammatory mediators in lung epithelial cells and in monocytes. VIP and PACAP prevented in monocytes the SARS-CoV-2-induced activation of NF-kB and SREBP1 and SREBP2, transcriptions factors involved in proinflammatory reactions and lipid metabolism, respectively. They also promoted CREB activation, a transcription factor with antiapoptotic activity and negative regulator of NF-kB. Specific inhibition of NF-kB and SREBP1/2 reproduced the anti-inflammatory, antiviral and cell death protection effects of VIP and PACAP. Our results support further clinical investigations of these neuropeptides against COVID-19.

microbiology↗

bioRxiv2020

The in vitro antiviral activity of the anti-hepatitis C virus (HCV) drugs daclatasvir and sofosbuvir against SARS-CoV-2

Current approaches of drugs repurposing against 2019 coronavirus disease (COVID-19) have not proven overwhelmingly successful and the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic continues to cause major global mortality. Daclatasvir (DCV) and sofosbuvir (SFV) are clinically approved against hepatitis C virus (HCV), with satisfactory safety profile. DCV and SFV target the HCV enzymes NS5A and NS5B, respectively. NS5A is endowed with pleotropic activities, which overlap with several proteins from SARS-CoV-2. HCV NS5B and SARS-CoV-2 nsp12 are RNA polymerases that share homology in the nucleotide uptake channel. We thus tested whether SARS-COV-2 would be susceptible these anti-HCV drugs. DCV consistently inhibited the production of infectious SARS-CoV-2 in Vero cells, in the hepatoma cell line (HuH-7) and in type II pneumocytes (Calu-3), with potencies of 0.8, 0.6 and 1.1 M, respectively. Although less potent than DCV, SFV and its nucleoside metabolite inhibited replication in Calu-3 cells. Moreover, SFV/DCV combination (1:0.15 ratio) inhibited SARS-CoV-2 with EC50 of 0.7:0.1 M in Calu-3 cells. SFV and DCV prevented virus-induced neuronal apoptosis and release of cytokine storm-related inflammatory mediators, respectively. Both drugs inhibited independent events during RNA synthesis and this was particularly the case for DCV, which also targeted secondary RNA structures in the SARS-CoV-2 genome. Concentrations required for partial DCV in vitro activity are achieved in plasma at Cmax after administration of the approved dose to humans. Doses higher than those approved may ultimately be required, but these data provide a basis to further explore these agents as COVID-19 antiviral candidates.

microbiology↗

bioRxiv2019

Deep learning-based methods for individual recognition in small birds

O_LIIndividual identification is a crucial step to answer many questions in evolutionary biology and is mostly performed by marking animals with tags. Such methods are well established but often make data collection and analyses time consuming and consequently are not suited for collecting very large datasets. C_LIO_LIRecent technological and analytical advances, such as deep learning, can help overcome these limitations by automatizing data collection and analysis. Currently one of the bottlenecks preventing the application of deep learning for individual identification is the need of hundreds to thousands of labelled pictures required for training convolutional neural networks (CNNs). C_LIO_LIHere, we describe procedures that improve data collection and allow individual identification in captive and wild birds and we apply it to three small bird species, the sociable weaver Philetairus socius, the great tit Parus major and the zebra finch Taeniopygia guttata. C_LIO_LIFirst, we present an automated method that allows the collection of large samples of individually labelled images. Second, we describe how to train a CNN to identify individuals. Third, we illustrate the general applicability of CNN for individual identification in animal studies by showing that the trained CNN can predict the identity of birds from images collected in contexts that differ from the ones originally used to train the CNNs. Fourth, we present a potential solution to solve the issues of new incoming individuals. C_LIO_LIOverall our work demonstrates the feasibility of applying state-of-the-art deep learning tools for individual identification of birds, both in the lab and in the wild. These techniques are made possible by our approaches that allow efficient collection of training data. The ability to conduct individual identification of birds without requiring external markers that can be visually identified by human observers represents a major advance over current methods. C_LI

ecology↗

bioRxiv2019

How to make methodological decisions when inferring social networks

Constructing and analysing social networks data can be challenging. When designing new studies, researchers are confronted with having to make decisions about how data are collected and networks are constructed, and the answers are not always straightforward. The current lack of guidance on building a social network for a new study system might lead researchers to try several different methods, and risk generating false results arising from multiple hypotheses testing. We suggest an approach for making decisions when developing a network without jeopardising the validity of future hypothesis tests. We argue that choosing the best edge definition for a network can be made using a priori knowledge of the species, and testing hypotheses that are known and independent from those that the network will ultimately be used to evaluate. We illustrate this approach by conducting a pilot study with the aim of identifying how to construct a social network for colonies of cooperatively breeding sociable weavers. We first identified two ways of collecting data using different numbers of feeders and three ways to define associations among birds. We then identified which combination of data collection and association definition maximised (i) the assortment of individuals into breeding groups (birds that contribute towards the same nest and maintain cohesion when foraging), and (ii) socially differentiated relationships (more strong and weak relationships than expected by chance). Our approach highlights how existing knowledge about a system can be used to help navigate the myriad of methodological decisions about data collection and network inference.\n\nSIGNIFICANCE STATEMENTGeneral guidance on how to analyse social networks has been provided in recent papers. However less attention has been given to system-specific methodological decisions when designing new studies, specifically on how data are collected, and how edge weights are defined from the collected data. This lack of guidance can lead researchers into being less critical about their study design and making arbitrary decisions or trying several different methods driven by a given preferred hypothesis of interest without realising the consequences of such approaches. Here we show that pilot studies combined with a priori knowledge of the study species social behaviour can greatly facilitate making methodological decisions. Furthermore, we empirically show that different decisions, even if data are collected under the same context (e.g. foraging), can affect the quality of a network.

animal behavior and cognition↗

bioRxiv2019

Anthropogenic Soils Promote Biodiversity in Amazonian Rainforests

Amazonian Dark Earths (ADEs) are fertile anthropic soils found throughout Amazonia, resulting from long-term occupation by pre-Columbian societies. Although the chemistry of these soils is well known, their biodiversity, particularly soil invertebrate communities have been neglected. To address this, we characterised soil macroinvertebrate communities and their activities in ADEs, comparing them with adjacent reference soils under forests and agriculture, at nine archaeological sites. We found 667 morphospecies and a tenacious pre-Columbian biodiversity footprint, with 40% of species found exclusively in ADEs. Soil biological activity was higher in ADEs than in adjacent soils, and associated with higher biomass and richness of organisms known to engineer the ecosystem. We show that these habitats have unique species pools, but that contemporary land-use causes nutrient loss and threatens their diversity. Both past and present human activities alter biodiversity and its distribution in Amazonia, and further efforts are needed to recognize and preserve these ADEs and their biodiversity.

zoology↗