Glycogen is the most effective energy reserve for metabolism in aquatic shellfish, and also contributes to the flavor and quality of oyster. Jinjiang oyster Crassostrea ariakensis is an economically and ecologically important species in China. In this study, RNA sequencing (RNA-seq) and assay for transposase-accessible chromatin using sequencing (ATAC-seq) were performed respectively to explore the gene expression and dynamic changes of chromatin accessibility among the oysters with different glycogen contents. A total of 9483 differentially expressed genes (DEGs) and 7215 significantly differential chromatin accessibility genes (DCAGs) were obtained, with the intersection of DEGs and DCAGs reaching 2600. Many of those genes were enriched in the pathways related to glycogen metabolism such as "Glycogen metabolic process" and "Starch and sucrose metabolism". In addition, a total of 526 SNP loci associated with glycogen content obtained by the genome wide association study (GWAS) corresponded to 241 genes, 63 of which were also DEGs and DCAGs as revealed above. This study will enrich basic research data and provide insights into the molecular mechanisms underlying the regulation of glycogen metabolism in oyster.
Widespread species that live in diversified environments have large population sizes and a high capacity for environmental tolerance and thus for range expansion. In contrast, narrow-ranged species are confined in restricted geographical areas and ecologically adjusted to narrow environmental conditions and consequently may have limited capacity to expand into a novel environment. However, it is not clear how genomic mechanisms underlie the differentiation in the closely related species that differ in their distribution ranges. The Niviventer niviventer species complex (NNSC), a group of the most abundant wild rats in Southeast Asia and China, provides an ideal system to investigate these questions, as this group includes widespread and narrow-ranged species that are phylogenetically closely related. In the present study, we combined ecological niche models with phylogenetic analysis suggesting that sister species cannot be both widespread and dominant in the same geographical region. Heterozygosity, linkage disequilibrium decay and Tajima's D analysis suggested that widespread species have higher genetic diversity than narrow-ranged species. Moreover, scanning ‘genomic islands of speciation’ revealed 13 genes in highly divergent regions that were shared by two widespread species, which distinguished them from the narrow-ranged species. In addition, functional annotation results suggested that these genes are involved in nervous system development and regulation. Adaptive evolution of these genes likely plays an important role in the speciation of these widespread species.
Monitoring the prevalence of the antimicrobial resistance gene (ARG) is critical to address the global crisis of antibiotic resistant bacterial infections. However, the characterization of ARG and microbiome structure, as well as the indicators for routine ARG monitoring in pig farms, are still lacking under the variation in antimicrobial exposure between countries/regions. Hence, metagenomics and Random Forest machine learning algorithm were used to decipher the ARG profiles, microbiome, and ARG contamination indicators in pig manure under different antimicrobial pressures between China and Europe. The results showed that Chinese pigs exposed to high level antimicrobials had higher total and plasmid-mediated ARG abundances than European pigs (P < 0.05). ANT(6)-Ib, APH(3')-IIIa, and tet(40) were the shared core ARGs between Chinese and European pigs. The core ARG identified in pigs existed a linkage between corresponding country/regions pigs and humans. Moreover, Lactobacillus and Prevotella were the dominant phyla in the core microbiome of Chinese and European pigs, respectively. Forty ARG markers and 43 biomarkers were found to differentiate Chinese and European pig manure samples with 100% and 98.7% accuracy, respectively. We identified indicators to assess the ARG contamination in Chinese and European pigs with high accuracy (r = 0.72 ~ 0.88). Escherichia flexneri in Chinese and European pigs carried numerous ARGs, ranging from 21 to 37. This study emphasized the importance of global collaboration in reducing antimicrobial resistance (AMR) risk and provided indicators for evaluating the risk of ARG contamination in pig farms.
Patterns of competition and cooperation in environment affect animal societies. In folivorous colobine primates, group size is predicted to increase in year-round food abundant rainforests due to less feeding competition. However, the observed pattern shows larger groups in seasonally leaf-deprived high-altitude/latitude montane ecosystems. This paradox is hypothesized to arise from cooperative benefits in heterogeneous environments. To test this, we first performed 6-year fieldwork on two neighboring groups of the golden snub-nosed monkey (Rhinopithecus roxellana), which is the northernmost distributed colobine primate. We found the group adjusted movement and selected high-quality habitats in response to fluctuating climate and spatial-temporally heterogeneous resources indicating a dynamic foraging strategy. In winter with cold temperatures and scarce resources, the large group inhabited food-rich habitats but didn't show significantly longer daily travel distances than its neighboring small group. We then compiled an eco-behavioral dataset of 52 colobine species to determine the evolutionary path. One path suggests that an increase in group size may have resulted from an enlargement of the home range in response to the cold and heterogeneous climates in high altitudes or latitudes. Hence, we introduce a multi-benefits framework to interpret large group formation integrating the environment heterogeneity. In cold, heterogeneous environments, even small groups require extra-large home ranges for dynamic survival needs. The spatiotemporal specificity of high-quality resources within the enlarged home range allows for frequent group encounters, which promotes social aggregation into larger groups by increasing the benefits of collective action and reproduction while constraining travel costs through a dynamic foraging strategy.
Parkinson's disease (PD) is a neurodegenerative condition that results in dyskinesia. This movement disorder is commonly influenced by oxidative stress, which suggests that antioxidant peptides may hold potential for PD treatment. In this study, a novel Cathelicidin peptide named Cath-KP (GCSGRFCNLFNNRRPGRLTLIHRPGGDKRTSTGLIYV) was identified from the skin of Kaloula pulchra frog. Circular dichroism and homology modeling analyses demonstrated that Cath-KP possesses a unique αββ structure, while in vitro experiments using ABTS and DPPH radical scavenging activities and FRAP analysis confirmed its antioxidant properties. Additionally, experiments utilizing the MPP+-induced dopamine neuron cell line and MPTP-induced PD mice found that Cath-KP can be internalized into cells and delivered to deep brain tissues, resulting in improved cell viability and the prevention of oxidative stress damage by promoting the expression of antioxidant enzymes and alleviating the accumulation of mitochondrial and intracellular ROS through activating Sirt1/Nrf2 pathway. FAK and p38 also play a part in the regulation of it. Ultimately, Cath-KP administration to MPTP-induced PD mice resulted in the restoration of the quantity of tyrosine hydroxylase (TH) - positive neurons and TH contents, and improved dyskinesia as observed in behavioral experiments. To our knowledge, this is the first Cathelicidin to demonstrate potent antioxidant and neuroprotective properties in PD models by targeting oxidative stress. These findings expand the known functions of Cathelicidins and hold promise for the development of therapeutic agents for PD.
The intestinal tract has critical roles in digesting and absorbing foods, expelling remaining wastes, and defensing against microorganisms. Profiling single cell transcriptome of intestinal track has greatly enriched the understandings on the cell types and their corresponding function heterogeneity that are important for intestine track development and disease. Although single cell transcriptome of the intestine tracts was extensively investigated in human and mice, the single cell gene expression profiles of pig caecum remained unexplored. Here, we performed single-cell RNA-seq on 45,572 cells from seven caecum samples of pigs at four developmental stages at day 30, 42, 150 and 730. We uncovered 12 major cell types and 38 subtypes, and characterized their featured genes, transcription factors and regulons, many of which were conserved in humans. We showed expansion of relative proportions of CD8+ T, GZMA (low) NKT, and decreasing of epithelium stem cells, Treg, RHEX+ T and plasmacytoid dendritic cells along the four developmental stages. We noted the upregulation of mitochondrial genes COX2 and ND2, as well as genes involved immune activation in multiple cell types after weaning. Cell-cell crosstalk analysis uncovered IBP6+ fibroblasts acted as the main signal senders at D30 while at other stages the IBP6- fibroblasts took over this role. T/NK cells interacted with epithelial cells and IBP6+ fibroblasts using GZMA-F2RL1/F2RL2 pairs only in D730 caecum. The present study provided important knowledge on cell type heterogeneity and function of pig caecum along different development stages.
T cells response in mammals requires synergism of the first signal and co-stimulatory signal. However, whether and how dual signaling regulates the T-cell response in early vertebrates remains unknown. In present study, we found that the teleost Nile tilapia Oreochromis niloticus encodes the key components of LAT signalosome: LAT, ITK, GRB2, VAV1, SLP-76, GADS, and PLC-γ1. These components are evolutionarily conserved, and CD3ε mAb-induced T-cell activation markedly increased their expression; while at least ITK, GRB2, and VAV1 could interact with LAT to form the signalosome. Downstream of the first signal, the NF-κB, MAPK/Erk, and PI3K-AKT pathways were activated upon CD3ε mAb stimulation. Furthermore, treatment of lymphocytes with CD28 mAbs triggered the AKT-mTORC1 pathway downstream of the co-stimulatory signal. A combined stimulation using CD3ε plus CD28 mAbs enhanced the phosphorylation of Erk1/2 and S6 and elevated the expression of NFAT1, c-Fos, IL-2, CD122, and CD44, signifying T-cell activation. Moreover, instead of the first signal or co-stimulatory signal alone, both dual signals are required for T-cell proliferation. In addition, full T-cell activation was accompanied by marked apoptosis and cytotoxic response. Therefore, our findings suggest that tilapia relies on dual signaling to maintain the optimal T-cell response, providing a novel perspective for understanding the evolution of the adaptive immune system.
Since the outbreak of COVID-19, many SARS-CoV-2-related coronaviruses (CoVs) have been discovered. Previous research identified a novel lineage of SARS-CoV-2-related CoVs in bat, such as RsYN04, which recognizes human angiotensin-converting enzyme 2 (ACE2) and poses a potential threat to humans. Here, we screened the binding of RsYN04 receptor-binding domain (RBD) to ACE2 orthologs from 52 animal species, and found that the virus shows a narrower ACE2-binding spectrum than SARS-CoV-2. However, the T484W mutation in the RBD can broaden the spectrum. We also evaluated 44 SARS-CoV-2 antibodies targeting seven epitope communities in RBD, together with the sera from COVID-19 convalescents and vaccinees, for their cross-reaction against RsYN04. Our results showed that all the antibodies, except for RBD-6 and RBD-7 classes, did not bind to RsYN04 RBD, indicating the substantial immune difference from SARS-CoV-2. Furthermore, the complex structure of the RsYN04 RBD with a cross-reactive antibody S43 in RBD-7 revealed a potently broad epitope for the development of therapeutics and vaccines. Our findings suggest RsYN04 and other viruses belonging to the same clade have the potential to infect several species including humans, highlighting the necessity for viral surveillance and development of broad anti-coronavirus countermeasures.
PINK1 is believed to be a mitochondrial kinase that phosphorylates Parkin and other proteins, playing a crucial role in mitophagy and protecting against neurodegeneration. Mutations in PINK1 and Parkin lead to loss of function and early onset of Parkinson's disease. However, there is a lack of strong in vivo evidence in rodent models to support the theory that loss of PINK1 affects mitophagy and induces neurodegeneration. Additionally, PINK1 knockout pigs have not been reported to exhibit neurodegeneration. Our recent work on non-human primates demonstrates that PINK1 is selectively expressed in primate brains, but not in rodent brains. To extend this finding to other species, such as pigs, we used multiple antibodies to examine the expression of PINK1 in pig tissues. Compared to monkey tissues, we were unable to obtain convincing data showing detectable PINK1 protein expression in pig tissues. Knocking down PINK1 in cultured pig cells did not result in altered phosphorylation of Parkin and BAD, as observed in cultured monkey cells. A comparison of monkey and pig striatum revealed that there were more PINK1-phosphorylated substrates in the monkey brain. Consistently, knocking out PINK1 in pigs did not lead to obvious changes in the phosphorylation of Parkin and BAD. These findings provide new evidence that PINK1 expression is specific to primates and underscores the importance of using non-human primates to investigate the function of PINK1 and the pathology related to PINK1 deficiency.
The gut microbiome interacts with the host to maintain body homeostasisits and dysbiosis features in many diseases. The underlying mechanisms of gut microbe regulation of host behavior and brain functions remain unclear. This study aimed to elucidate the influence of gut microbiota on brain functions via post-translational modifications mechanisms under the presence of bacteria and sterile state without any stimulation. We performed a succinylome analysis of hippocampal proteins in GF and SPF mice and metagenomics of feces from SPF mice. And integrated these results with previously reported hippocampal acetylome and phosphorylome data from the same batch. Then followed by bioinformatics analyses. 584 succinylation sites on 455 proteins were identified, including up-regulation of 54 succinylation sites on 91 proteins and down-regulation of 99 sites on 51 proteins in GF compared to SPF. We constructed a panoramic map of gut microbiota-regulated succinylation, acetylation, and phosphorylation, and identified the “cross-talk” and relative independence between different types of post-translational modifications in modulating complicated intracellular pathways. A Pearson correlation analysis indicated that 13 taxa, mainly belonging to Bacteroidetes, correlated with the biological functions of post-translational modifications. Positive correlations between these taxa and succinylation and negative correlations with acetylation were identified in modulating the intracellular pathways. This is the first report of hippocampal physiological changes induced by the absence of gut microbiota and proteomic quantification of succinylation, phosphorylation and acetylation, which contributes to understanding the involvement of the gut microbiome in brain function and behavioral phenotypes.
Changes in protein abundance and in reversible protein phosphorylation (RPP) play important roles in regulating hypometabolism but have never been documented in overwintering frogs at high altitudes. To test the hypothesis that protein abundance and protein phosphorylation change in response to winter hibernation, we conducted a comprehensive and quantitative proteomic and phosphoproteomic analysis of the liver of the Xizang plateau frog, Nanorana parkeri, living on the Qinghai-Tibet Plateau. In total, 5170 proteins were quantified and 5695 phosphorylation sites on 1938 proteins were quantified, respectively. Proteomic results showed that a total of 674 (438 up, 236 down) differentially expressed proteins were screened in hibernating N. parkeri versus summer individuals. Functional enrichment analysis revealed that highly expressed proteins in winter were significantly enriched in immune-related signaling pathways, whereas low-expressed proteins were mainly involved in metabolic processes. A total of 4251 (4147 up, 104 down) modified sites belonging to 1638 (1555 up, 83 down) phosphoproteins showed a significant change in the liver. A large set of proteins are regulated during hibernation by RPP and these proteins are involved in various functions such as many metabolic enzymes, ion pumping, protein turnover, signal transduction, and alternative splicing. These changes contribute to enhancing protection, suppressing energy-consuming processes, and thus metabolic depression. Moreover, phosphofructokinase, glutamate dehydrogenase, and ATPase activities were all significantly lower in winter compared to summer. In conclusion, our present results support the hypothesis and demonstrate the importance of RPP as a regulatory mechanism when animals transition into a hypometabolic state.
Geographical backgrounds and dispersal ability might have strong imprint on assemblage dissimilarity; however, these aspects have generally been overlooked in large-scale beta diversity studies. Here, we examined whether patterns and drivers of taxonomic beta diversity (TBD) and phylogenetic beta diversity (PBD) of breeding birds in China vary across 1) regions on both sides of the Hu Line, a line that demarcates China’s topographical, climatic, economic, and social patterns, and 2) species with different dispersal ability. TBD and PBD were calculated and partitioned into turnover and nestedness components using a moving window approach. Variables representing climate, habitat heterogeneity, and habitat quality were used to evaluate the effects of environmental filtering, whereas spatial distance was used to assess the influences of dispersal limitation. Variance partitioning analysis was used to assess the relative role of these variables. In general, TBD and PBD values were high in mountainous areas and environmental filtering largely determined TBD and PBD. However, different dominating environmental filters on both sides of the Hu Line led to divergent beta diversity patterns. Specifically, climate-driven species turnover and habitat heterogeneity-related species nestedness dominated the regions at east and west of the Hu Line, respectively. Additionally, bird species with stronger dispersal ability were more susceptible to environmental filtering resulting in more homogeneous assemblages. Our results indicated that regions with distinctive geographical backgrounds might present different ecological factors that lead to divergent assemblage dissimilarity patterns, and dispersal ability determines the response of assemblages to these ecological factors. Identifying a single universal explanation for the observed pattern without considering these aspects might lead to simplistic or incomplete conclusions. Therefore, it is essential to consider the combined effect of geographical background and dispersal ability for comprehensively understanding large-scale patterns of beta diversity and for planning conservation strategies.
Omega-3 polyunsaturated fatty acids (n-3 PUFAs), particularly docosahexaenoic acid (22:6n-3, DHA), have been shown to play a crucial role in reproduction and reproductive health of vertebrates, including humans. However, the underlying mechanism of this phenomenon remains largely unknown. In this study, we used two zebrafish genetic models, the elovl2-/- mutant as an endogenous DHA-deficient model and the fat1 (an omega-3 desaturase encoding gene) transgenic zebrafish as an endogenous DHA-rich model, to investigate the effects of DHA on oocyte maturation and oocyte quality. Our results show that the elovl2-/- mutant zebrafish had much lower fecundity and poorer oocyte quality than the wildtype controls, while the fat1 zebrafish had higher fecundity and better oocyte quality than wildtype controls. DHA deficiency in elovl2-/- embryos led to defects of egg activation, poor microtubule stability and reduced pregnenolone levels. Further study reveals that DHA promotes pregnenolone synthesis by promoting the transcription of cyp11a1, which encodes the cholesterol side-chain cleavage enzyme, thereby stabilizing microtubule assembly during oogenesis. In turn, the hypothalamic–pituitary–gonadal (HPG) axis is enhanced by DHA. In conclusion, using two unique genetic models, our study demonstrates the endogenously synthesized DHA promotes oocyte maturation and oocyte quality by promoting pregnenolone production via transcriptional regulation of cyp11a1.
The Recalcitrance of pathogens to traditional antibiotics has made treating and eradicating bacterial infections more difficult. In this regard, developing new antimicrobial agents to combat antibiotic-resistant strains has become a top priority. Antimicrobial peptides (AMPs), a ubiquitous class of naturally occurring compounds with broad-spectrum anti-pathogen activity, hold significant promise as one of the most effective solutions to the current antimicrobial resistance (AMR) crisis. Several AMPs have been identified and evaluated for their therapeutic application, with many already in the drug development pipeline. Their distinct properties, such as high target specificity, potency, and the ability to bypass microbial resistance mechanisms, make them a promising alternative to traditional antibiotics. Nonetheless, several challenges, such as high toxicity, lability to proteolytic degradation, low stability, poor pharmacokinetics, and high production costs, continue to hamper their clinical applicability. Therefore, much recent research has focused on optimizing antimicrobial peptides’ properties to improve their performance. By understanding the physiochemical properties of AMPs, such as amphipathicity, hydrophobicity, structural conformation, amino acid distribution, and composition, that correspond to their activity, researchers can design AMPs with desired and improved performance. In this review, we highlight some of the key strategies, rational design and de novo design, used to optimize the performance of AMPs. We also discuss how the evolution of predictive computational tools and technologies that utilize artificial intelligence and machine learning are increasingly being used to design and synthesize lead drug candidates with high efficacy.
LncRNAs (long non-coding RNAs) have been recognized as key modulators in mammalian immunity, and particular emphasis is placed on the mechanism of lncRNA-mediated ceRNA (competitive endogenous RNA) crosstalk. To date, investigations on lncRNAs in lower vertebrates still remain tentative. In the present study, we characterized the first immune-related lncRNA, termed pol-lnc78, in teleost fish Japanese flounder (Paralichthys olivaceus). We found that pol-lnc78 acts as a ceRNA for pol-miR-n199-3p to SARM (sterile alpha and armadillo motif-containing protein), the last discovered TIR adaptor. This intertwined ceRNA trinity regulates flounder antibacterial responses through TLR signaling pathway. Specifically, SARM as a negative regulator exacerbates bacterial infection through inhibiting the expression of inflammatory cytokines IL-1β and TNF-α. Pol-miR-n199-3p reduces SARM expression via specific interacting with the 3’UTR region, thereby promoting SARM-dependent inflammatory cytokines expression and protecting host against bacterial dissemination. Furthermore, pol-lnc78 sponges pol-miR-n199-3p to relieve the expression inhibition on SARM. In particular, during infection, the negative regulators pol-lnc78 and SARM are significantly downregulated, while pol-miR-n199-3p is significantly upregulated in expression, thus favoring host antibacterial defense. The results of this study provide new insights into the mechanism of fish immunity, and open up new horizons to a better understanding of ceRNA crosstalk in lower vertebrates.
Regulatory sequences and transposable elements (TEs) account for a large proportion of the genome sequences of species, while their roles in gene transcription, especially tissue-specific expression, remain largely unknown. Pigs are an excellent animal models for studying the biology of the genome sequences owing to their great diversity of wild and domesticated populations. Here, we integrated H3K27ac ChIP-seq, H3K4me3 ChIP-seq and RNA-seq data from 10 tissues of the same 7 fetuses and their consanguineously related 8 adult pigs to annotate the regulatory elements and TEs for their links with histone modifications and mRNA expression across varying tissues and development stages. The association analyses of mRNA expression with H3K27ac and H3K4me3 peak activity revealed that H3K27ac peaks showed stronger associations with gene expression than H3K4me3. We revealed that 1.45% of the TEs overlapped with H3K27ac or H3K4me3 peaks, of which the majority displayed tissue-specific activity. We identified a TE subfamily (LTR4C_SS) with binding motifs for SIX1 and SIX4 that showed specific enrichment in adult and fetal ovary H3K27ac peaks. We also revealed widespread expression of TEs as part of exons or promoters of genes from the RNA-seq data, including 4688 TE-containing transcripts that displayed development stage and tissue-specific expression. Notably, 1967 TE-containing transcripts were enriched in the testes. We highlighted that an LTR acting as a testis-specific alternative promoter in SRPK2 (a cell cycle-related protein kinase) in our pig data, MLT1F1, was also conserved in humans and mice, suggesting an ancient embedding of the TEs in testis-specific expressed genes or parallel evolution. Collectively, our work demonstrates that TEs are deeply embedded in the genome and exhibit important tissue-specific biological functions, particularly in the reproductive organs.
A total of 10 Alcyonacea corals were collected at depths ranging from 900 m to 1640 m by the manned submersible Shenhai Yongshi during two cruises in the South China Sea (SCS). Phylogenetic distance and average nucleotide identity (ANI) analyses of mitochondrial genomes combined with morphology examination and sclerite scanning showed that the collected samples could be assigned to four suborders - Calcaxonia, Holaxonia, Scleraxonia, and Stolonifera - which might represent 10 novel deep-sea species. The analyses of the dissimilarity of GC skew, phylogenetic distance, and ANI indicated that the evolution of Octocorallia mitochondrial sequences was slow. The nonsynonymous (Ka) and synonymous (Ks) substitution (Ka/Ks) ratios indicated that 14 protein-coding genes (PCGs) were undergoing purifying selection and that the selection pressures might be from specific deep-sea environments. The correlation analysis of median values of Ka/Ks ratio of five gene families and environmental factors showed that the genes encoding cytochrome b (cob) and DNA mismatch repair protein (mutS) might be driven by environmental factors to format deep-sea species. This study highlighted the slow evolution and adaptative mechanism of deep-sea corals in the deep ocean.
Non-alcoholic fatty liver disease (NAFLD) has been widely reported to have a potential association with LBP (Lipopolysaccharide-binding protein) mutation. However, the mechanisms, especially epigenetic mechanisms underpinning the association remain elusive. Herein, we constructed LBP-/- rats with NAFLD and performed integrative analysis of targeting-active enhancer H3K27ac chromatin immunoprecipitation coupled with high-throughput sequencing and transcriptomic sequencing to explore the potential epigenetic pathomechanisms of active enhancer in the exacerbation of NAFLD upon LBP deficiency. Intriguingly, we found that LBP-/- could reduce inflammatory response but markedly deteriorate HFD-induced NAFLD in rats, with abundant alterations of histone acetylome and regulatory transcriptome. In total, 1,128 differential enhancer-target genes significantly enriched in cholesterol metabolic process and fatty acid metabolic process were identified with | Cor (peak-gene correlation) | > 0.5 and a | log2 (fold change) | > 1.5 between WT and LBP-/- NAFLD rats. Notably, based on our integrative analysis method, we screened out TF C/EBPβ (CCAAT/enhancer-binding protein β) as a pivotal contributor to dysregulated histone acetylome H3K27ac and lipid metabolism gene SCD as a downstream effector to exacerbate NAFLD. Thus, this study not only broadens our understanding of the essential role of LBP in the pathogenesis of NAFLD from the perspective of epigenetics, but identifies key TF C/EBPβ and hub functional gene SCD as potential regulators that may serve as possible therapeutic targets.

Estimation of population size is a core of wildlife population ecology and conservation. Although GPS collars has long been considered as an effective and promising method on monitoring behavior of wildlife, few studies have used this technique to estimate wildlife population size. In this study, we used the GPS telemetry method for conducting population census. We deployed GPS collars on 32 rhesus macaques (Macaca mulatta) in 9 groups on a small island and collected 109,739 fixes from May 2021 to April 2022. We used fixes of all tracked individuals within a group to calculate home range for each group. The annual home range of 9 groups ranged from 13.9 ha to 55.4 ha. Home ranges had substantial overlap with neighboring groups, and the largest ratio of exclusively use occurred in March. We estimated that the study area had the capacity to accommodate 38.6 macaque groups within their respective home ranges and the population size was 957 ± 193 macaques on the island by multiplying the mean group size (24.8 ± 5.0). In addition, we compared the similarity of utilization distribution between each individual and their belonging group. The result indicated that using a single individual’s dataset could represent the whole group’s home range and utilization pattern on most occasions. Our method is labor saving, independent of experience and observational ability of surveyors, of high repeatability, and can generate a huge amount of fixes to explore other important ecological questions.

Quantification of behaviors in macaques provides crucial support for various scientific disciplines, including pharmacology, neuroscience, and ethology. Despite recent advancements in the analysis of macaque behavior, research on multi-label behavior detection in socially housed macaques, including consideration of interactions among them, remains scarce. Given the lack of relevant approaches and datasets, we developed the Behavior-Aware Relation Network (BARN) for multi-label behavior detection of socially housed macaques. Our approach models the relationship of behavioral similarity between macaques, guided by a behavior-aware module and novel behavior classifier, which is suitable for multi-label classification. We also constructed a behavior dataset of rhesus macaques using ordinary RGB cameras mounted outside their cages. The dataset included 65 913 labels for 19 behaviors and 60 367 proposals, including identities and locations of the macaques. Experimental results showed that BARN significantly improved the baseline SlowFast network and outperformed existing relation networks. In conclusion, we successfully achieved multi-label behavior detection of socially housed macaques with both economic efficiency and high accuracy.