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ABSTRACT There are complex interactions between an organism's microbiome and its response to stressors, often referred to as the ‘gut–brain axis’; however, the ecological relevance of this axis in wild animals remains poorly understood. Here, we used a chronic mild stress protocol to induce stress in wild-caught house sparrows (Passer domesticus), and compared microbial communities among stressed animals, those recovering from stress, captive controls (unstressed) and a group not brought into captivity. We assessed changes in microbial communities and abundance of shed microbes by culturing cloacal samples on multiple media to select for aerobic and anaerobic bacteria and fungi. We complemented this with cultivation-independent 16S and ITS rRNA gene amplification and sequencing, pairing these results with host physiological and immune metrics, including body mass change, relative spleen mass and plasma corticosterone concentrations. We found significant effects of stress and captivity on the house sparrow microbiomes, with stress leading to an increased relative abundance of endotoxin-producing bacteria – a possible mechanism for the hyperinflammatory response observed in captive avians. While we found evidence that the microbiome community partially recovers after stress cessation, animals may lose key taxa, and the abundance of endotoxin-producing bacteria persists. Our results suggest an overall link between chronic stress, host immune system and the microbiome, with the loss of potentially beneficial taxa (e.g. lactic acid bacteria), and an increase in endotoxin-producing bacteria due to stress and captivity. Ultimately, consideration of the host's microbiome may be useful when evaluating the impact of stressors on individual and population health. 

ABSTRACT At any given time, only a subset of microbial community members are active in their environment. The others are in a state of dormancy, with strongly reduced metabolic rates. It is of interest to distinguish active and inactive microbial cells and taxa to understand their functional contributions to ecosystem processes and to understand shifts in microbial activity in response to change. Of the methods used to assess microbial activity-dormancy dynamics, 16S rRNA/rRNA gene amplicons (16S ratios) and active cell staining with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) are two of the most common, yet each method has limitations. Given that in situ activity-dormancy dynamics are proxied only by laboratory methods, further study is needed to assess the level of agreement and potential complementarity of these methods. We conducted two experiments investigating microbial activity in plant-associated soils. First, we treated corn field soil with phytohormones to simulate plant soil stress signaling, and second, we used rhizosphere soil from common bean plants exposed to drought or nutrient enrichment. Overall, the 16S ratio and CTC methods exhibited similar patterns of relative activity across treatments when treatment effects were large, and the instances in which they differed could be attributed to changes in community size (e.g., cell death or growth). Therefore, regardless of the method used to assess activity, we recommend quantifying community size to inform ecological interpretation. Our results suggest that the 16S ratio and CTC methods report comparable patterns of activity that can be applied to observe ecological dynamics over time, space, or experimental treatment. IMPORTANCE Although the majority of microorganisms in natural ecosystems are dormant, relatively little is known about the dynamics of the active and dormant microbial pools through both space and time. The limited knowledge of microbial activity-dormancy dynamics is in part due to uncertainty in the methods currently used to quantify active taxa. Here, we directly compared two of the most common methods (16S ratios and active cell staining) for estimating microbial activity in plant-associated soil and found that they were largely in agreement in the overarching patterns. Our results suggest that 16S ratios and active cell staining provide complementary information for measuring and interpreting microbial activity-dormancy dynamics in soils. They also support the idea that 16S rRNA/rRNA gene ratios have comparative value and offer a high-throughput, sequencing-based option for understanding relative changes in microbiome activity, as long as this method is coupled with quantification of community size. 

Nutrient enrichment impacts ecosystems globally. Population history, especially past resource environments, of numerically dominant plant species may affect their responses to subsequent changes in nutrient availability. Eutrophication can also alter plant–microbe interactions via direct effects on associated microbial communities or indirect effects on dominant species’ biomass production/allocation as a result of modified plant–soil interactions.

We combined a greenhouse common garden and a field reciprocal transplant of a salt marsh foundation species (Spartina alterniflora) within a long‐term, whole‐ecosystem, nutrient‐enrichment study to determine whether enrichment affects plant production and microbial community structure differently depending on plant population history. For the greenhouse portion, we collected 20S. alternifloragenotypes—10 from an enriched creek that had received elevated nutrient inputs for 10 years and 10 from an unenriched reference creek—and reared them in a common garden for 1 year. For the field portion, we conducted a 2‐year, fully crossed reciprocal transplant experiment with two gardens each at the enriched and unenriched sites; we examined the effects of source site (i.e. population history), garden site and plant genotype.

After 2 years, plants in enriched gardens had higher above‐ground biomass and altered below‐ground allocation compared to plants in unenriched gardens. However, performance also depended on plant population history: plants from the enriched site had decreased above‐ground and rhizome production compared to plants from the unenriched site, most notably in unenriched gardens. In addition, almost all above‐ and below‐ground traits varied depending on plant genotypic identity.

Effects of nutrient enrichment on the associated microbial community were also pronounced. Following 1 year in common garden, microbial community structure varied by plant population history andS. alternifloragenotypic identity. However, at the end of the reciprocal transplant, microbial communities differed primarily between enriched and unenriched gardens.

Synthesis. Nutrient enrichment can impact plant foundation species and associated soil microbes in the short term. Most importantly, nutrient enrichment can also have long‐lasting effects on plant populations and associated microbial communities that potentially compromise their ability to respond to changing resource conditions in the future.