The precise targeted integration of large exogenous genes into chicken sex chromosomes is of particular interest for sex-linked trait manipulation, sex-control breeding, and the development of avian bioreactor models. However, efficient targeted integration at sex chromosome loci remains technically challenging, and optimized editing strategies for these loci are still lacking. To improve targeted genome editing at two previously identified chicken sex chromosome safe-harbor loci, EE0.6 and NC_006127.4, this study systematically evaluated and optimized key parameter affecting editing efficiency. First, we evaluated the effects of different sgRNA combinations on targeted knockout efficiency, establishing the advantage of a dual-sgRNA/Cas9 architecture, which achieved knockout efficiencies of 86.67% and 75.00% at the EE0.6 and NC_006127.4 loci, respectively. We next introduced the Cas9 nickase (Cas9n) system, which has previously been reported to exhibit improved editing specificity, and evaluated its performance at both target loci. Quantitative analysis showed that the dual-sgRNA/Cas9 targeting system successfully mediated the precise targeted integration of a 1.1-kb SV40-mCherry reporter cassette, reaching 100% (28/28) at the EE0.6 locus and 80.00% (20/25) at the NC_006127.4 locus. Based on this result, this study further investigated the effects of donor homology arm (HA) lengths (200 bp, 600 bp, and, 1000 bp) and vector topologies (circular and linearized) on targeted knock-in efficiency. The results revealed that in the circular donor system, the optimal HA lengths for the EE0.6 and NC_006127.4 loci were 200 bp (50.4% ± 4.4%) and 600 bp (30.1% ± 1.2%), respectively. However, upon the introduction of linearized donors with free ends, the knock-in efficiency of the exogenous target fragment was significantly enhanced, and its HA length preference underwent a significant reversal. The optimal HA length for EE0.6 was extended to 600 bp (78.9% ± 1.0%), whereas that for NC_006127.4 was shortened to 200 bp (49.9% ± 0.4%). In summary, this study established an efficient targeted integration strategy for chicken sex chromosome loci. The optimized system provides a foundation for future applications in sex-linked breeding and avian bioreactor development.

Tannic acid (TA) is a hydrolysable plant secondary metabolite known to influence multiple physiological processes in animals; however, its role in regulating brown adipose tissue (BAT) thermogenesis remains poorly understood. Notably, the overwinter food caches of Brandt’s voles predominantly consist of Artemisia species, which are rich in TA. This study aimed to determine whether TA contributes to cold tolerance in Brandt’s voles by activating BAT thermogenesis. Adult male voles were administered TA, after which the masses of BAT and inguinal white adipose tissue (iWAT) were measured, and temperature changes in BAT, the body surface, and the rectum were recorded following exposure to − 20 °C. In addition, transcriptomic analyses of BAT were performed, and the expression and protein levels of key thermogenic markers were assessed. The results showed that TA reduced iWAT mass while exerting minimal effects on BAT mass. TA-treated voles exhibited significantly elevated temperatures in BAT, the body surface, and the rectum after cold exposure. Histological analyses revealed that TA treatment reduced adipocyte area in iWAT while increasing the number of nuclei in brown adipocytes in BAT. In BAT, differentially expressed genes (DEGs) in voles receiving a low TA dose were significantly enriched in pathways related to fat digestion and absorption and peroxisome proliferator-activated receptor (PPAR) signaling. In contrast, DEGs in voles administered a high TA dose were predominantly associated with brown adipocyte differentiation and the upregulation of cold-induced thermogenesis. Moreover, TA administration increased the expression of FFAR4 and UCP1, as well as the protein levels of PGC-1α, PPARγ, and UCP1 following cold exposure. Collectively, these findings demonstrate that TA enhances cold tolerance in Brandt’s voles by promoting thermogenic gene expression and stimulating brown adipocyte differentiation in BAT, providing novel insights into the role of plant secondary metabolites in mammalian cold adaptation and herbivore–plant interactions.

Adolescent stress not only exerts enduring effects on individual behavior and physiology, but also shapes offspring phenotypes through mechanisms of transgenerational inheritance. However, the transgenerational effects of predation risk, an important ecological factor, remain poorly characterized in wild, social rodents. Using male Brandt's voles (Lasiopodomys brandtii) as a model, this study systematically investigated the transgenerational effect of adolescent exposure to cat urine, rabbit urine (as a non-predator stimulus), and distilled water (as a control) for 60 min daily over 18 consecutive days on the behavioral and physiological traits of future offspring. Our data showed that while male paternal adolescent cat odor (CO) exposure did not significantly alter overall parental investment by either parent, it induced significant phenotypic changes in offspring. These included an increased female ratio at weaning and reduced post-weaning weight gain. Furthermore, adolescent offspring exhibited decreased locomotor activity in the open field test, while adult offspring displayed heightened vigilant rearing and reduced head-out behavior when confronted with CO exposure. These behavioral and developmental alterations were accompanied by elevated serum levels of adrenocorticotropic hormone and corticosterone, suggesting enhanced basal activity of the hypothalamic-pituitary-adrenal axis. These findings demonstrate that male paternal experience with predation risk during adolescence can transgenerationally regulate the sex ratio, growth and development, stress response, and antipredator strategies of future offspring, independent of alterations in parental investment in wild, social rodents. This study elucidates the unique role of the paternal lineage in the transgenerational inheritance of early adversity and provides further experimental evidence for understanding how environmental stress drives adaptive phenotypic transmission across generations.

Sex determination is a crucial process in animal development, regulated by complex genetic networks. In avian species, Doublesex and mab-3 related transcription factor 1 (DMRT1) plays a vital role in gonadal development and sex determination. To unravel the function of DMRT1 in chicken sex determination, establishing an inducible DMRT1 knockout model is essential. In this study, we constructed an inducible DMRT1 knockout system and verified its efficiency and effects on related genes and physiological indicators. To achieve precise genomic ablation, we screened multiple sgRNAs targeting the DMRT1 locus and integrated the optimal sequence into a doxycycline-responsive (Tet-on) CRISPR/Cas9 architecture. For in vitro experiments, vectors were delivered via cell transfection and induced with 20 µg mL-1 doxycycline (DOX), achieving an 80% knockout efficiency. Following the administration of polyethylenimine (PEI)-encapsulated plasmids into chicken embryos, we successfully implemented the inducible system in vivo. Quantitative analysis confirmed a mosaic knockout of DMRT1 with an observed efficiency reaching 45%. Following targeted disruption, we evaluated sex-related gene and protein expression alterations via qRT-PCR and Western blot (WB). Furthermore, ELISA was performed to measure testosterone levels in male embryonic gonads across multiple developmental stages (E4.5 to E18.5). qRT-PCR analysis showed that after induction, female-related genes (CYP19A1, FOXL2, ESR1) were significantly upregulated, and male-related genes (DMRT1, SOX9, AMH) were significantly downregulated. WB results revealed increased protein expression levels of CYP19A1 and FOXL2, and decreased protein expression of SOX9 post-induction. ELISA confirmed that testosterone levels in the gonads of induced male embryos were significantly reduced compared to normal and non-induced males. The study successfully established an inducible DMRT1 knockout system in chickens. This system effectively regulates the expression of sex-related genes and reduces testosterone levels in male embryos, providing theoretical and technical support for breeding novel sex-controlled breeding materials.

Coprophagy, the ingestion of feces, is widespread among vertebrates and plays an important role in hindgut-fermenting herbivores by facilitating nutrient recycling and maintaining gut microbial stability. However, most studies have focused on cecal or fecal microbiota, with limited attention to microbial communities at other body sites. Here, we used Brandt's vole, a species exhibiting frequent coprophagic behavior, to examine how coprophagy prevention affects hindgut fermentation and microbial communities across multiple sites, including the tongue, lung, stomach, and cecum. Coprophagy prevention increased food intake but reduced body weight gain and significantly decreased cecal acetate, propionate, and butyrate concentrations. At the microbial level, coprophagy prevention increased the Simpson index of the tongue microbiota and induced modest compositional shifts in the tongue, lung, and stomach microbiota. In contrast, the cecal microbiota exhibited greater compositional and predicted functional changes. Notably, coprophagy prevention reduced the Bray-Curtis distance between tongue and cecal microbiota without altering the number of shared amplicon sequence variants. Together, these findings indicate that coprophagy not only promotes efficient nutrient recycling through hindgut fermentation but may also influence microbiota composition across behaviorally interconnected digestive niches.

Herbivores significantly influence plant communities by modifying interspecies relationships, which in turn impacts ecosystem functioning. Although excluding herbivores is expected to enhance the dominance of perennial plants, few studies have consistently explored how plants adjust the growth-defense-reproduction trade-off in response to changes at different stages of herbivore exclusion. We conducted a controlled fence experiment in Inner Mongolia grassland to examine the effects of vole exclusion on the dominance and adaptation strategies of two perennial grasses. We found that twelve years of Brandt's vole grazing altered the plant community composition, but in the second year of the exclusion experiment, Leymus chinensis quickly regained its dominance. This rapid recovery was facilitated by L. chinensis strategically shifting its growth-defense-reproduction investment over time. In the early exclusion phase, L. chinensis quickly enhanced its competitive edge by prioritizing vegetative growth and clonal reproduction. As the exclusion period progressed, the species transitioned to seed-based dispersal to enhance population fitness. In contrast, the dominance of Cleistogenes squarrosa was largely influenced by interspecific interactions rather than intrinsic adaptive changes. These results reveal plants can dynamically adjust their resource investment strategies to optimize population fitness at different stages of vegetation recovery. This enhances our understanding of plant community dynamics and the establishment and maintenance of dominant species in grassland ecosystems.

Predator cues have profound impacts on the behavior and physiology of prey animals. However, the mechanisms underlying stress responses induced by chronic exposure to predator cues in mammals, particularly the role of the gut microbiota, remain insufficiently understood. This study investigated how gut microbiota contributes to behavioral and physiological responses in Brandt's voles (Lasiopodomys brandtii) under chronic predator odor exposure. Adult voles were repeatedly exposed to cat feces odor (CO) for 18 days (1 h/day), followed by behavioral tests to assess anti-predator and anxiety-like behaviors, hormonal measurements to evaluate basal hypothalamic-pituitary-adrenal (HPA) axis activity, and 16S rRNA sequencing to analyze gut microbiota composition. The results showed that repeated CO exposure increased anxiety-like behaviors and basal HPA axis activity in both sexes. However, anti-predator behaviors exhibited sex-specific responses: Males were habituated to repeated CO exposure by reducing freezing and alerting behaviors, whereas females consistently displayed concealing strategies, reflecting different adaptive strategies to prolonged predator threats. While CO exposure did not alter the α-diversity of gut microbiota, it significantly affected the β-diversity in females, including a reduction in the relative abundance of Treponema and Quinella. Cecal microbiota transplantation from female CO-exposed voles to naive recipients increased anxiety-like behaviors and basal HPA levels but did not alter anti-predator behaviors in the recipients. In contrast, male recipients showed no significant behavioral or physiological changes. These findings suggest that gut microbiota is involved in regulating anxiety-like behavior and HPA axis activity in female voles but has a limited regulating role in male voles.

The acute activation of serotonin 1 A (5-HT1A) receptors appears to disrupt maternal behavior in rats; however, the underlying neuroanatomical mechanisms remain poorly understood. We employed two approaches to investigate the role of 5-HT1A receptors in maternal behavior to address this knowledge gap. First, we used real-time polymerase chain reaction (PCR) to analyze 5-HT1A receptor mRNA expression in female rats at different reproductive stages. We identified stage- and region-specific expression patterns, including temporary increases in the nucleus accumbens (NAc), ventral tegmental area (VTA), and dorsal raphe nucleus (DRN), as well as a temporary decrease in the medial prefrontal cortex (mPFC), amygdala, hippocampus, and ventromedial hypothalamic nucleus (VMH) during the perinatal, early, and middle postpartum periods. These findings suggest that coordinated 5-HT1A activity across these brain regions is critical for normal maternal behavior. Second, we used c-Fos immunohistochemistry to elucidate the central mechanisms underlying the effects of the acute and repeated administration of 8-OH-DPAT (a 5-HT1A receptor agonist, 1.0 mg/kg, sc.) on maternal behavior. Acute 8-OH-DPAT administration disrupted maternal behaviors, including pup retrieval, pup licking, nest building and hovering over pups, while simultaneously increased c-Fos expression in the mPFC, ventral bed nucleus of the stria terminalis (vBNST), NAc shell, lateral septum (LS), and medial amygdala (MeA). Disruptions in pup retrieval, pup licking and nest building persisted following five days of repeated 8-OH-DPAT treatment, whereas hovering over pups showed substantial recovery, returning to near-normal levels. Concurrently, c-Fos expression increased in the vBNST but decreased in the mPFC, MeA, and DRN. These results suggest that acute and repeated 8-OH-DPAT administration disrupts maternal behavior via distinct presynaptic and postsynaptic 5-HT1A receptor mechanisms. This study highlights the complex regulatory role of 5-HT1A receptor activity in maternal care and provides insights into the neuroanatomical and neurochemical mechanisms underlying maternal behavior.

Prenatal maternal stress negatively impacts maternal mental health and mother-child interaction, potentially increasing the risk of developmental outcomes in offspring. While studies in humans and lab animals have established these associations, the underlying mechanisms in naturalistic settings, where stressors are dynamic and interspecies differences may emerge, remain poorly characterized. This study introduced a novel model system using wild Brandt’s voles (Lasiopodomys brandtii) to investigate the effects of maternal depression-like psychopathology on offspring mood, cognition, and brain development. This model involves the repeated exposure of pregnant voles to predator odors, a natural stressor, which induces a depression-like state from late pregnancy to the early postpartum period. This model integrates ecologically relevant stressors with neurobehavioral assessments in a wild-derived species, allowing for a mechanistic investigation in a biologically meaningful context. We found that while exposure to predator odor stress during pregnancy induced maternal depressive-like states, it did not alter the level of postnatal parent-offspring interaction. Offspring born to mothers exposed to predator odor during pregnancy exhibited increased anxiety- and depression-like behaviors, impaired spatial and social cognition, and reduced sociability compared to offspring of mothers exposed to distilled water. These offspring also showed reduced neurogenesis in the hippocampal dentate gyrus, along with decreased dendrite branching and spine density. Our findings suggest that the effects of depression-like states during pregnancy and postpartum in female Brandt’s voles on offspring brain and behavioral functions occur independently of parent-offspring interactions, with hippocampal structural and functional abnormalities potentially mediating behavioral deficits. Importantly, this work establishes Brandt’s voles as a new, ecologically valid animal model for studying gestational depression and its intergenerational outcomes, bridging the gap between laboratory rodent studies and natural behavioral contexts.

Aromatase, a crucial enzyme for estrogen synthesis, plays a vital role in gender determination and differentiation. This study aimed to establish an inducible knockout model of the chicken CYP19A1 gene, which encodes aromatase, to support gender control in chickens. We selected the most efficient sgRNA target site and constructed an inducible knockout model based on the Tet-on system. The knockout efficiency reached 80% with 20 μg/mL DOX induction in vitro. The encapsulation of the plasmid with PEI and injection into eggs achieved a knockout efficiency of 45% in ovo. qRT-PCR analysis revealed a significant downregulation of female-related genes (CYP19A1, FOXL2, ESR1) and upregulation of male-related genes (DMRT1, SOX9, AMH) in female chicken embryos after induction. Western blotting showed decreased protein expression of CYP19A1 and FOXL2, and increased SOX9 expression in female embryos post-DOX induction. Elisa detection further confirmed lower estradiol levels in the gonads of induced female embryos compared to normal and non-induced females. These findings demonstrate the successful establishment of an inducible knockout system for the CYP19A1 gene in chickens, providing theoretical and technical support for the creation of new breeding materials for gender control.

Environmental stressors encountered during pregnancy can exert profound effects on fetal development and program long-term physiological outcomes in offspring. Among these, ecologically relevant stressors such as predation risk remain understudied despite their potential to disrupt maternal physiology and placental function. In this study, we investigated how prenatal exposure to predator odor influences placental glucocorticoid metabolism and fetal growth in Brandt's voles (Lasiopodomys brandtii). Pregnant voles were exposed to predator odor to simulate prenatal predator stress, allowing us to assess its effects on fetal growth, maternal and fetal corticosterone levels, and the expression of key enzymes and transport proteins involved in glucocorticoid regulation. The results revealed that prenatal exposure to predator odor significantly reduced fetal weight. While maternal and fetal serum corticosterone levels increased, placental corticosterone levels remained unchanged. Additionally, our study observed significant increases in the expression of placental P-glycoprotein and 11β-hydroxysteroid dehydrogenase 1, both of which are crucial for maintaining placental glucocorticoid metabolic homeostasis. However, 11β-hydroxysteroid dehydrogenase 2 levels were unaffected. In the fetal brain, corticotropin-releasing hormone expression in the hypothalamus showed a downward trend, whereas glucocorticoid receptor expression in the hippocampus remained unchanged, indicating a disruption in the development of the hypothalamic-pituitary-adrenal axis. These findings suggest that prenatal exposure to predator odor alters placental glucocorticoid metabolism, leading to increased fetal corticosterone exposure and potentially impairing fetal development.

Tannic acid (TA), a representative plant secondary metabolite, impairs intestinal immune function and alters intestinal microbiota abundance of Brandt's voles. Whether TA reduces intestinal immune function by modifying the gut microbiota remains unknown. In this study, Brandt's voles were divided into three groups for microbiota transplantation (FMT): normal saline (Ab), microbiota from normal saline administration (R-Con), and microbiota from TA administration (R-TA). Then, the intestinal morphology, immune indices, gut flora, and microbiota metabolites were measured after FMT. The findings showed that the sIgA content of small intestine of voles in the R-TA group were lower than in the R-con group. Additionally, the voles in the R-con group exhibited higher mRNA levels of PIgR, J-chain, BAFF, and APRIL than in the R-TA group. The ANOSIM results showed significant structural differentiation, reflecting that the β-diversity of the cecal microbiome was altered. Moreover, the voles in the R-con group had a higher concentration of butyric acid in the cecum compared to both the Ab and R-TA groups. In the experiment 2, the sIgA content in the duodenum and ileum of Brandt's voles in the TBA group (1200 mg•kg-1d-1 TA + butyrate) was significantly higher than that in the TNS group (1200 mg•kg-1d-1 TA + normal saline). To summarize, the "TA microbiota" decreased the slgA secretion and synthesis in the small intestine by reducing butyric acid content, thus lowering the intestinal immune capacity of Brandt's voles. The findings provide the experimental basis to prove the critical role of gut microorganisms in controlling animal physiological processes.

The impact of burdock tea (BT) made from burdock (Arctium lappa) roots in normal individuals and animal models remains largely unknown, particularly on lung protection. This study examined responses of oxidative stress, inflammation, and the microbiota within the cecum and the lung to BT treatment in healthy Wistar rats. A middle-dose BT reduced the Chao1 and Shannon indices, and both low and middle doses induced structural alterations in the cecal microbiota. Additionally, low doses increased the abundances of Phascolarctobacterium, Alloprevotella, Desulfovibrio, and the NK4A214 group. In the lung, middle and high doses increased Corynebacterium, with high doses also boosting Megasphaera and Lactobacillus. Functionally, low doses downregulated the biosynthesis of antibiotics in the cecal microbiota, while middle doses reduced the Epstein–Barr virus and Escherichia coli pathogenic infection pathways; additionally, middle and high doses modulated chromosomal proteins and bile acid biosynthesis in the pulmonary microbiota. BT treatment enhanced the content of short-chain fatty acids (SCFAs), upregulated the expression of GPR43, and suppressed NLRP3 expression in both the colon and lung tissues, while concurrently promoting the expression of ZO-1 and Occludin. Furthermore, serum levels of IL-1β and IL-6, as well as tissue levels of MDA, were significantly reduced. Notably, propionate exhibited an inverse correlation with MDA, IL-6, and NLRP3, while showing a positive correlation with ZO-1. Similarly, acetate was negatively correlated with MDA and NLRP3 and positively correlated with ZO-1. Overall, BT exhibits a nontoxic profile and may protect lung tissue through its antioxidant nature and gut–lung axis mediated by SCFAs.

Paternal predation risk can program offspring phenotypes via maternal responses and epigenetic marks of spermatozoa. However, the processes and consequences of this experience in biparental species are unknown. Here, we examined how preconception and postconception paternal cat odor (CO) exposure affects anxiety-like behavior and antipredator response in Brandt's voles (Lasiopodomys brandtii). We found that preconception paternal CO exposure inhibited maternal investment when offspring were raised by mothers alone, while postconception exposure increased paternal investment towards the offspring raised by both parents. The increased paternal behavior may be associated with an increasing grooming behavior received from their mates, which alleviated the anxiety-like behavior in CO-exposed males. Both paternal experiences increased the levels of anxiety-like behavior in adolescent offspring but differentially altered adult phenotypes. Specifically, adult females from preconception CO-exposed fathers spent less time in defensive concealing, whereas the offspring of postconception CO-exposed fathers showed more in response to acute cat urine exposure. Correspondingly, baseline corticosterone levels were decreased and increased in these offspring, respectively. Our results indicate that in biparental species, paternal predation risk exposure affects offspring phenotypes in pathway-dependent and age-specific manners and that only the presence of both parents can elicit adaptive responses to a high predation-risk environment.

Sound and stocking density are two common factors which influence the performance and welfare of layers. Accumulated studies have been conducted on the impacts of the two factors on production performance, while knowledge regarding the impacts of the two factors and their interactions on stress-related serum indicators, behaviors, and cecal bacterial communities in laying hens is still limited. A 3 × 3 factorial design with three sound sources (natural sound (NS), instrumental music (IMS), or mixed road noise (MRS)) and three stocking densities (low density (LD), medium density (MD), and high density (HD)) was used in this 24-day experiment, in which 378 30-week-old Xiandao green-shell layers were randomly distributed into nine treatments with six replicates per treatment. At the 3rd, 12th, and 24th experimental day, we evaluated the serum levels of adrenocorticotropic hormone (ACTH) and corticosterone (CORT) and recorded stress-related animal behaviors. At the end of the experiment, 16S rRNA gene amplicon sequencing of the cecal bacterial communities was performed. Our results confirmed that MRS and HD induced significantly elevated serum ACTH and CORT levels, and were correlated with significantly increased feather pecking behavior. IMS and LD were associated with enhanced preening behavior and reduced feather pecking behavior. LD significantly increased the Firmicutes/Bacteriodetes ratio and IMS significantly enriched the beneficial Lactobacillus population. Based on the obtained results we proposed that music exposure and reduced stocking density were helpful in reducing stress and improving cecal bacterial profile, which were beneficial for improving layers’ health status and welfare.

AbstractThe gut microbiota of rodents is essential for survival and adaptation and is susceptible to various factors, ranging from environmental conditions to genetic predispositions. Nevertheless, few comparative studies have considered the contribution of species identity and geographic spatial distance to variations in the gut microbiota. In this study, a random sampling survey encompassing four rodent species (Apodemus agrarius, Cricetulus barabensis, Tscherskia triton and Rattus norvegicus) was conducted at five sites in northern China's farming–pastoral ecotone. Through a cross‐factorial comparison, we aimed to discern whether belonging to the same species or sharing the same capture site predominantly influences the composition of gut microbiota. Notably, the observed variations in microbiome composition among these four rodent species match the host phylogeny at the family level but not at the species level. The gut microbiota of these four rodent species exhibited typical mammalian characteristics, predominantly characterized by the Firmicutes and Bacteroidetes phyla. As the geographic distance between populations increased, the number of shared microbial taxa among conspecific populations decreased. We observed that within a relatively small geographical range, even different species exhibited convergent α‐diversity due to their inhabitation within the same environmental microbial pool. In contrast, the composition and structure of the intestinal microbiota in the allopatric populations of A. agrarius demonstrated marked differences, similar to those of C. barabensis. Additionally, geographical environmental elements exhibited significant correlations with diversity indices. Conversely, host‐related factors had minimal influence on microbial abundance. Our findings indicated that the similarity of the microbial compositions was not determined primarily by the host species, and the location of the sampling explained a greater amount of variation in the microbial composition, indicating that the local environment played a crucial role in shaping the microbial composition.

Rodents, including the striped field mouse (Apodemus agrarius), play vital roles in ecosystem functioning, with their gut microbiota contributing significantly to various ecological processes. Here, we investigated the structure and function of 94 wild A. agrarius individuals from 7 geographic populations (45°57′ N, 126°48′ E; 45°87′ N, 126°37′ E; 45°50′ N, 125°31′ E; 45°59′ N, 124°37′ E; 46°01′ N, 124°88′ E; 46°01′ N, 124°88′ E; 46°01′ N, 124°88′ E), revealing two distinct enterotypes (Type1 and Type2) for the first time. Each enterotype showed unique microbial diversity, functions, and assembly processes. Firmicutes and Bacteroidetes dominated, with a significant presence of Lactobacillus and Muribaculaceae. Functional analysis highlighted metabolic differences, with Type1 emphasizing nutrient processing and Type2 showing higher energy production capacity. The analysis of the neutral model and the null model revealed a mix of stochastic (drift and homogenizing dispersal) and deterministic processes (homogenous selection) that shape the assembly of the microbiota, with subtle differences in the assembly processes between the two enterotypes. Correlation analysis showed that elevation and BMI were associated with the phylogenetic turnover of microbial communities, suggesting that variations in these factors may influence the composition and diversity of the gut microbiota in A. agrarius. Our study sheds light on gut microbial dynamics in wild A. agrarius populations, highlighting the importance of considering ecological and physiological factors in understanding host–microbiota interactions.

Abstract Rodents can cause considerable changes in plant community composition. However, relationships between shifts in species dominance and plant functional traits caused by rodents have seldom been investigated, especially for belowground functional traits. In this study, a set of enclosures was constructed to analyze the effects of 10 years of Brandt's voles' activities on the defense strategies and dominant position changes of three gramineous plants (Leymus chinensis, Stipa krylovii, and Cleistogenes squarrosa) in Inner Mongolia. Here, we measured the dominance, biomass, and fourteen functional traits of three plants. The effects of Brandt's voles on dominance, biomass, and functional traits were analyzed, and then we explored the effect of functional traits on plant dominance by using the structural equation model. Results showed that long‐term feeding by Brandt's voles resulted in a significant decrease in the dominance of L. chinensis and S. krylovii, whereas C. squarrosa was positively affected. The belowground biomass of L. chinensis and S. krylovii was higher in the vole treatment, which showed that they were increasing their escape characteristics. The leaf thickness of L. chinensis and the leaf C:N ratio of S. krylovii significantly increased, while the specific leaf area of C. squarrosa significantly decreased. All three gramineous showed increased resistance traits in response to Brandt's voles, which positively affected their dominance. Tolerance‐related traits of S. krylovii significantly increased, with the increasing growth rate of root length contributing to enhancing its dominance. We highlight that selective feeding by rodents led to the selection of different defense strategies by three gramineous plants, and that changes in biomass allocation and functional traits in the different species affected plant dominance, driving changes in the plant communities.

6-methoxybenzoxazolinone (6-MBOA) is a secondary plant metabolite predominantly found in monocotyledonous plants, especially Gramineae. In damaged tissue, 2-β-D-glucopyranosyloxy-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA-Glc) is hydrolyzed to DIMBOA, which spontaneously decomposes into 6-MBOA. It is commonly detected in plants consumed by voles and livestock and can also be present in cereal-based products. Discovered in 1955, this compound is renowned for its ability to trigger animal reproduction. However, there is a lack of research on its functional and mechanistic properties, leaving much of their potential unexplored. This review aimed to comprehensively summarize the effects of 6-MBOA on animal reproduction and human health, as well as its defensive role against herbivores. Studies have shown that 6-MBOA effectively inhibits the digestion, development, growth, and reproduction of insects. 6-MBOA may act as a partial agonist of melatonin and exert a regulatory role in mammalian reproduction, resulting in either promoting or inhibiting effects. 6-MBOA has been theorized to possess anti-tumor, anti-AIDS, anti-anxiety, and weight-loss effects in humans. However, insufficient attention has been paid to its defense properties against mammalian herbivores, and the mechanisms underlying its effects on mammalian reproduction remain unclear. In addition, research on its impact on human health is still in its preliminary stages. The review emphasizes the need for further systematic and comprehensive research on 6-MBOA to fully understand its diverse functions. Elucidating the effects of 6-MBOA on animal reproduction, adaptation, and human health would advance our understanding of plant–herbivore coevolution and the influence of environmental factors on animal population dynamics. Furthermore, this knowledge could potentially promote its application in human health and animal husbandry.

Food waste is a common issue arising from grinding of food by experimental animals, leading to excessive food scraps falling into cages. In the wild, animals grind food by gnawing vegetation and seeds, potentially damaging the ecological environment. However, limited ecology studies have focused on food grinding behavior since the last century, with even fewer on rodent food grinding, particularly recently. Although food grinding’s function is partially understood, its biological purposes remain under-investigated and driving factors unclear. This review aims to explain potential causes of animal food grinding, identify influencing factors, and discuss contexts and limitations. Specifically, we emphasize recent progress on gut microbiota significance for food grinding. Moreover, we show abnormal food grinding is determined by degree of excess normal behavior, emphasizing food grinding is not meaningless. Findings from this review promote comprehensive research on the myriad factors, multifaceted roles, and intricate evolution underlying food grinding behavior, benefiting laboratory animal husbandry and ecological environment protection, and identifying potential physiological benefits yet undiscovered.