Distinct Gut Microbial Enterotypes and Functional Dynamics in Wild Striped Field Mice (Apodemus agrarius) across Diverse Populations.

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.

Geographical distribution and species variation of gut microbiota in small rodents from the agro-pastoral transition ecotone in northern China.

The 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.

Grassland biodiversity response to livestock grazing, productivity, and climate varies across biome components and diversity measurements.

Livestock overgrazing and climate change have been identified as the primary causes of grassland degeneration and biodiversity decline, yet the underlying mechanism remains unclear. To gain a better understanding of this, we conducted a meta-analysis of 91 local or regional field studies from 26 countries across all inhabited continents. Using concise statistical analyses, we assessed five theoretical hypotheses for grazing intensity, grazing history, grazing animal type, productivity, and climate, and decomposed the individual contributions of each factor in regulating multiple components of grassland biodiversity. After controlling for confounding effects, we found that: no significant linear or binomial pattern for the effect-size of grassland biodiversity as grazing intensity increased; the effect-size of producer richness was relatively lower (negative biodiversity response) in grasslands with a short grazing history, grazed by large livestock, high productivity, or high climate suitability; additionally, significant difference for the effect-size of consumer richness was only detected across grazing animal groups; and the effect-size of consumer abundance, and decomposer abundance all displayed significant differences with respect to grazing characters, grassland productivity, and climate suitability. Besides, results of hierarchical variance partitioning suggested that the total and individual contribution of predictors varied across biome components and diversity measurements. Specifically, grassland productivity acted as a key factor in driving producer richness. The findings presented here collectively suggest that the response of grassland biodiversity to livestock grazing, productivity, and climate varies across different components of the biome and measurements of diversity.

Timing outweighs magnitude of rainfall in shaping population dynamics of a small mammal species in steppe grassland.

Climate change-induced shifts in species phenology differ widely across trophic levels, which may lead to consumer-resource mismatches with cascading population and ecosystem consequences. Here, we examined the effects of different rainfall patterns (i.e., timing and amount) on the phenological asynchrony of population of a generalist herbivore and their food sources in semiarid steppe grassland in Inner Mongolia. We conducted a 10-y (2010 to 2019) rainfall manipulation experiment in 12 0.48-ha field enclosures and found that moderate rainfall increases during the early rather than late growing season advanced the timing of peak reproduction and drove marked increases in population size through increasing the biomass of preferred plant species. By contrast, greatly increased rainfall produced no further increases in vole population growth due to the potential negative effect of the flooding of burrows. The increases in vole population size were more coupled with increased reproduction of overwintered voles and increased body mass of young-of-year than with better survival. Our results provide experimental evidence for the fitness consequences of phenological mismatches at the population level and highlight the importance of rainfall timing on the population dynamics of small herbivores in the steppe grassland environment.

Host-microbiota interaction helps to explain the bottom-up effects of climate change on a small rodent species.

The population cycles of small rodents have puzzled biologists for centuries. There is a growing recognition of the cascading effects of climate change on the population dynamics of rodents. However, the ultimate cause for the bottom-up effects of precipitation is poorly understood, from a microbial perspective. Here, we conducted a precipitation manipulation experiment in the field, and three feeding trials with controlled diets in the laboratory. We found precipitation supplementation facilitated the recovery of a perennial rhizomatous grass (Leymus chinensis) species, which altered the diet composition and increase the intake of fructose and fructooligosaccharides for Brandt's vole. Lab results showed that this nutrient shift was accompanied by the modulation of gut microbiota composition and functional pathways (especially for the degradation or biosynthesis of L-histidine). Particularly, the relative abundance of Eubacterium hallii was consistently increased after feeding voles with more L. chinensis, fructose or fructooligosaccharide. These modulations ultimately increased the production of short chain fatty acids (SCFAs) and boosted the growth of vole. This study provides evidence that the precipitation pulses cascades through the plant community to affect rodent gut microbiome. Our results highlight the importance of considering host-microbiota interaction when investigating rodent population responses to climate change.

Dietary shifts influenced by livestock grazing shape the gut microbiota composition and co-occurrence networks in a local rodent species.

The collapse of large wild herbivores with replacement of livestock is causing global plant community and diversity shifts, resulting in altered food availability and diet composition of other sympatric small herbivores in grasslands. How diet shifts affect the gut microbiota of small mammals and whether these changes may translate into complex interactions among coexisting herbivores remain largely unknown. We conducted both a field experiment and a laboratory diet manipulation experiment to test whether sheep grazing induces a diet shift and thus alters the gut microbiota of a small rodent species living in grassland. We found that enclosures subjected to grazing were mostly dominated by Stipa krylovii (accounting for 53.6% of the total biomass) and that voles consumed significantly more S. krylovii and less Cleistogenes squarrosa in grazed enclosures. Voles in grazing enclosures exhibited significantly lower abundances of Firmicutes, higher abundances of Bacteroidetes and significantly lower measurements of alpha diversity. The microbiota from voles in the grazed enclosures had a smaller and more simplified co-occurrence network with relatively higher percentage of positive interactions. Analysis based on dietary clusters indicated that grazing-induced changes in diet composition contributed to the distinct gut microbial community of voles in enclosures. We verified our findings using laboratory experiments, in which voles were exclusively fed C. squarrosa (high carbohydrate, high fibre and high in secondary compounds), S. krylovii (low carbohydrate, low fibre and low in secondary compounds) or Leymus chinensis (nutritionally intermediate). We observed that the gut microbiota of voles changed with the three different diets, supporting the idea that the effects of sheep grazing on the gut microbiota of Brandt's voles may be related to grazing-induced diet shifts. Our results highlighted the negative effects of livestock grazing on small mammals in grassland via changes in plant community and gut microbiota of small mammals and help to better understand the cascading consequences of realistic scenarios of world-wide decline in large wild herbivores.

Effects of maternal stress induced by predator odors during gestation on behavioral and physiological responses of offspring in Brandt's vole (Lasiopodomys brandtii).

Maternal effects play a crucial role in regulating populations of small mammals and anti-predator behaviors of offspring. The present study investigated the effects of maternal stress induced by exposure to predator odors during gestation on the behavioral and physiological responses of offspring in Brandt's vole (Lasiopodomys brandtii). Effects included changes in growth, behaviors, hormone levels in serum, and fosB/ΔfosB mRNA and FosB/ΔFosB protein expression in the hypothalamus. Our results showed that when pregnant voles were exposed to cat and rabbit urine odors for 18 days (1 h/day), the weight of the female offspring decreased; however, the thymus and ovary indices increased, compared with the control group. Male offspring of voles exposed to cat odor spent more time exploring and foraging, and showed a decrease in corticosterone (CORT) compared with voles in the control group. Both fosB/ΔfosB mRNA and FosB/ΔFosB protein expressions were downregulated in the hypothalamus of male offspring in the cat odor-exposed group compared to that in offspring in the control group. However, the mRNA levels of fosB/ΔfosB in the hypothalamus of female offspring did not decrease in response to cat urine odor. Our findings suggested that maternal stress induced by predator odors during gestation restrained the growth of female offspring and increased the duration of exploring and foraging behaviors of male offspring by regulating the HPA axis, which, in turn, reduced the sensitivity of offspring to unfamiliar environments and increased their probability of becoming prey.

Large manipulative experiments reveal complex effects of food supplementation on population dynamics of Brandt's voles.

Although food supplementation is well known to increase population density, there is still debate on the causative effects of food supplementation on reproduction, survival, and immigration. Large manipulative experiments, which exclude any confounding effects of dispersal and predation, are essential for clarifying the debate. In this study, we investigated the effects of food supplementation on Brandt's vole population dynamics and plant community in eight large enclosures (0.48 ha each) from 2010 to 2014. Food supplementation showed significant positive effects on population density due to increases in recruitment; however, it showed a complex effect on survival of voles: positive in non-breeding seasons, but negative in breeding seasons. In addition, food supplementation increased the quality of plants (as reflected by increased crude protein content), but decreased the quantity of less preferred plants in experimental enclosures. Thus, food seems to have direct positive effects on small rodents through improvement of food supply and indirect negative effects through food-induced density-dependent effects, and may have long-term effects on rodents through altering plant community composition and abundance.

Repeated exposure to cat urine induces complex behavioral, hormonal, and c-fos mRNA responses in Norway rats (Rattus norvegicus).

Prey species show specific adaptations that allow recognition, avoidance, and defense against predators. This study was undertaken to investigate the processing of a chronic, life-threatening stimulus to Norway rats (Rattus norvegicus). One hundred forty-four Norway rats were tested by repeated presentation of cat urine for 1 h at different days in a defensive withdrawal apparatus. Rats exposed to urine for short periods showed significantly larger defensive behavioral and medial hypothalamic c-fos messenger RNA (mRNA) responses than other groups. These defensive responses habituated shortly after the presentation of cat urine. Serum levels of adrenocorticotropic hormone and corticosterone increased significantly when animals were repeatedly exposed to cat urine. However, the hormonal responses took longer to habituate than the behavioral and molecular responses did. We conclude that the behavioral and c-fos mRNA responses are "primed" for habituation to repeated exposures to cat urine, while the hormonal responses show "resistance." The results support our hypothesis that the strongest anti-predator responses at three levels would occur during short-term exposure to cat urine and that these responses would subsequently disappear on prolonged exposure. This study assists understanding the way in which the different levels of defensive responses are integrated and react during chronic stress.

Successive sheep grazing reduces population density of Brandt's voles in steppe grassland by altering food resources: a large manipulative experiment.

Livestock grazing has shaped grassland ecosystems around the world. Previous studies indicated grazing showed various impacts on small rodents; however, most studies were conducted over 1-2 years without controlling for confounding factors such as immigration/emigration and predation in rodents. Brandt's voles (Lasiopodomys brandtii) are generally recognized as pests because of food overlap with domestic herbivores, but are also important for biodiversity conservation because they provide nests or food to many birds. Fully understanding the ecological relationship between domestic herbivores and small mammals is essential to making ecosystem management decisions. To address these needs, we carried out a field experiment during the period 2010-2013 to assess the effects of sheep grazing on vegetation and the population density of Brandt's voles along a gradient of three grazing intensities by using 12 large-scale enclosures. Responses of Brandt's voles to livestock grazing varied with grazing intensity and year. As compared to the control group, sheep grazing had no effect on vole abundance in the first year but an overall negative effect on vole abundance in the following 3 years. Successive grazing caused decreases in survival and male body mass of voles, but had no significant effect on fecundity. Negative effects of grazing were associated with a grazing-induced deterioration in both food quantity (reflected by biomass and cover of less-preferred plants), and food quality (measured by tannin and total phenol content). Our findings highlight the urgent need for more flexible management of yearly rotational grazing to optimize livestock production while maintaining species diversity and ecosystem health.

From Leaf Metabolome to In Vivo Testing: Identifying Antifeedant Compounds for Ecological Studies of Marsupial Diets.

Identifying specific plant secondary metabolites that influence feeding behavior can be challenging, but a solid understanding of animal preferences can guide efforts. Common brushtail possums (Trichosurus vulpecula) predominantly eat Eucalyptus species belonging to the subgenus Symphyomyrtus, and avoid eating those belonging to the Monocalyptus subgenus (also called subgenus Eucalyptus). Using an unbiased (1)H NMR metabolomics approach, a previous study identified unsubstituted B ring flavanones in most species of monocalypts examined, whereas these compounds were absent from symphyomyrtles. We hypothesised that unsubstituted B ring flavanones act as feeding deterrents for common brushtail possums. In the current study, we tested this hypothesis by comparing how much possums ate of a basal diet, with diets containing one of four structurally related compounds; pinocembrin, flavanone (unsubstituted B ring flavanones), chrysin (the flavone analogue of pinocembrin), and naringenin (a flavanone with B ring substitution). We found that pinocembrin and flavanone deterred feeding relative to the basal diet, but that chrysin and naringenin did not at equivalent concentrations. Thus, unsubstituted B-ring flavanones may explain why brushtail possums avoid eating monocalypt species. Furthermore, small differences in the structure of secondary compounds can have a large impact on antifeedant properties. These results demonstrate that metabolomics can be a valuable tool for ecologists seeking to understand herbivore feeding preferences.

Defensive responses of Brandt's voles (Lasiopodomys brandtii) to stored cat feces.

Predator odors are non-intrusive natural stressors of high ethological relevance. Animals are daily challenged with stressors of varying intensity and it is essential for their survival to respond to a wide range of threats. Behavioral and hormonal responses and changes in the level of medial hypothalamic c-fos mRNA were examined in Brandt's voles (Lasiopodomys brandtii) exposed to the feces of a domestic cat (Felis catus) stored for different periods. One hundred voles were tested in the defensive withdrawal apparatus. The voles showed an aversion to freshly collected cat feces, indicated by high levels of flight-related behaviors, increased freezing behavior, and more vigilant rearing compared to old feces. The serum levels of adrenocorticotropic hormone and corticosterone significantly increased when the voles were exposed to fresh cat feces. The level of c-fos mRNA in the medial hypothalamic region was highest in the individuals exposed to fresh cat feces. All of these behavioral, endocrine and c-fos-mRNA responses were lower when voles were subjected to older cat feces. We conclude that these responses depend on volatile chemical constituents of cat feces rather than their physical characteristics and that this accounts for the lower responses to feces stored for longer periods.

Defensive responses of Brandt's voles (Lasiopodomys brandtii) to chronic predatory stress.

Predator odors are non-intrusive natural stressors of high ethological relevance. The objective of this study was to investigate the processing of a chronic, life-threatening stimulus during repeated prolonged presentation to Brandt's voles. One hundred and twenty voles were tested by repeated presentation of cat feces in a defensive withdrawal apparatus. Voles exposed to feces for short periods showed more avoidance, more concealment in the hide box, less contact time with the odor source, more freezing behavior, less grooming, more jumping, and more vigilant rearing than did non-exposed voles, and those exposed for longer periods. Serum levels of adrenocorticotropic hormone and corticosterone increased significantly when animals were repeatedly exposed to cat feces for short periods. The behavioral and endocrine responses habituated during prolonged presentation of cat feces. ΔfosB mRNA expression level was highest in voles exposed to cat feces for 6 and 12 consecutive days, and subsequently declined in animals exposed to cat feces for 24 days. We therefore conclude that the behavioral and endocrine responses to repeated exposure to cat feces undergo a process of habituation, while ΔfosB changes in the medial hypothalamic region exhibit sensitization. We propose that habituation and sensitization are complementary rather than contradictory processes that occur in the same individual upon repeated presentation of the same stressor.

[Trophic niches of Pantholops hodgsoni, Procapra picticaudata and Equus kiang in Kekexili region].

Interspecies competition is the main factor determining the trophic niche width of sympatric species on both ecological and evolutionary levels. In August 2004, a microscopic analysis of feces was made to analyze the diet compositions of Pantholops hodgsoni, Procapra picticaudata and Equus kiang under free-living circumstances between Chuma River and Wudaoliang in the Kekexili National Nature Reserve. The trophic niche widths and overlaps of these herbivorous ungulates were calculated based on the diet compositions, and the results showed that these three ungulates had similar foods, but the proportions of their diet compositions were different. Gramineous plants were the main food sources for P. hodgsoni, P. picticaudata and E. kiang, accounting for 58.7%, 44.57% and 92.28% of the diet, respectively. The trophic niche widths of P. hodgsoni, P. picticaudata and E. kianthese were 0.878, 0.735 and 0.695, and the niche overlaps of P. hodgsoni and E. kiang, P. hodgsoni and P. picticaudata, and E. kiang and P. picticaudata were 0.869, 0.985 and 0.785, respectively, which suggested that there was a potential intense competition among them, especially for P. hodgsoni and P. picticaudata. The relations between competition and coexistence among P. hodgsoni, P. picticaudata and E. kiang were discussed from the viewpoints of their ecological characteristics and trophic ecology.