The Amargosa vole is a highly endangered rodent endemic to a small stretch of the Amargosa River basin in Inyo County, California. It specializes on a single, nutritionally marginal food source in nature. As part of a conservation effort to preserve the species, a captive breeding population was established to serve as an insurance colony and a source of individuals to release into the wild as restored habitat becomes available. The colony has successfully been maintained on commercial diets for multiple generations, but there are concerns that colony animals could lose gut microbes necessary to digest a wild diet. We analyzed feces from colony-reared and recently captured wild-born voles on various diets, and foregut contents from colony and wild voles. Unexpectedly, fecal microbial composition did not greatly differ despite drastically different diets and differences observed were mostly in low-abundance microbes. In contrast, colony vole foregut microbiomes were dominated by Allobaculum sp. while wild foreguts were dominated by Lactobacillus sp. If these bacterial community differences result in beneficial functional differences in digestion, then captive-reared Amargosa voles should be prepared prior to release into the wild to minimize or eliminate those differences to maximize their chance of success.

The critically endangered Amargosa vole (Microtus californicus scirpensis) is found only in rare marsh habitat near Tecopa, California in a plant community dominated by three-square bulrush (Schoenoplectus americanus). Since the earliest research on the Amargosa vole, the existing paradigm has been that these voles are obligatorily dependent on bulrush as their only food source and for the three-dimensional canopy and litter structure it provides for predator avoidance. However, no prior research has confirmed the diet of the Amargosa vole. In this study we characterized the Amargosa vole’ nutritional needs, analyzed the quality of bulrush by forage analysis, and performed microhistological and metabarcoding analyses of vole feces to determine what foods were consumed in the wild. All bulrush plant tissues analyzed were low in fat (from 0.9% of dry matter in roots to 3.6% in seeds), high in neutral detergent fiber (from 5.9% in rhizomes to 33.6% in seeds), and low in protein (7.3–8.4%). These findings support the conclusion that bulrush alone is unlikely to support vole survival and reproduction. Fecal microhistology and DNA metabarcoding revealed relatively diverse diets including plants in 14 families, with rushes (Juncaceae), bulrushes (Cyperaceae), and grasses (Poaceae) being the most common diet items. On microhistology, all analyzed samples contained bulrush, sedges (Carex sp.), rushes (Juncus sp.), and beaked spikerush (Eleocharis rostrellata) even from marshes where non-bulrush plants were uncommon. There was evidence of insects at <1% in two marshes but none in the remaining marshes. Metabarcoding detected ten genera of plants. When considering non-Schoenoplectus targets, for which metabarcoding had poor sensitivity, saltgrass (Distichlis spicata) was the most commonly detected species, with prominent contributions from seaside arrowgrass (Triglochin concinna) and yerba mansa (Anemopsis californica) as well. Diversity of vole diets generally increased with increasing site plant diversity, but differences were not statistically significant. Confirming details about dietary behaviors is critical for informing appropriate conservation planning including habitat management and reintroduction of voles into new sites.

A survey of the ecological variability within 52 populations of Schoenoplectus californicus (C.A. Mey.) Soják across its distributional range revealed that it is commonly found in nitrogen (N) limited areas, but rarely in phosphorus limited soils. We explored the hypothesis that S. californicus supplements its nitrogen demand by bacterial N₂-fixation processes associated with its roots and rhizomes. We estimated N₂-fixation of diazotrophs associated with plant rhizomes and roots from several locations throughout the species’ range and conducted an experiment growing plants in zero, low, and high N additions. Nitrogenase activity in rhizomes and roots was measured using the acetylene reduction assay. The presence of diazotrophs was verified by the detection of the nifH gene. Nitrogenase activity was restricted to rhizomes and roots and it was two orders of magnitude higher in the latter plant organs (81 and 2032 nmol C₂H₄ g DW^-1 d^-1, respectively). Correspondingly, 40x more nifH gene copies were found on roots compared to rhizomes. The proportion of the nifH gene copies in total bacterial DNA was positively correlated with the nitrogenase activity. In the experiment, the contribution of fixed N to the plant N content ranged from 13.8% to 32.5% among clones from different locations. These are relatively high values for a non-cultivated plant and justify future research on the link between N-fixing bacteria and S. californicus production.

While metapopulation theory offers tractable means to understand extinction risks for patchily-distributed endangered species, real systems often feature discrepant patch quality and accessibility, complex influences of environmental stochasticity, and regional and temporal autocorrelation. Spatially structured metapopulation models are flexible and can use real data but often at the cost of generality. Particularly as resources for management of such species are often critically limited, endangered species management guided by metapopulation modeling requires incorporation of biological realism. Here we developed a flexible, stochastic spatially structured metapopulation model of the profoundly endangered Amargosa vole, a microtine rodent with an extant population of only a few hundred individuals within 1km² of habitat in the Mojave Desert. Drought and water insecurity are increasing extinction risks considerably. We modelled subpopulation demographics using a Ricker-like model with migration implemented in an incidence function metapopulation model. A set of scenarios was used to assess the effect of anthropogenic stressors or management actions on expected time to extinction (T_e) including: 1) wildland fire, 2) anthropogenically-mediated losses of hydrologic flows, 3) drought, 4) intentional expansion of existing patches into ‘megamarshes’ (i.e. via restoration/enhancement), and 5) additive impacts of multiple influences. In isolation, marshes could be sources or sinks, but spatial context within the full metapopulation including adjacency could alter relative impacts of subpopulations on all other subpopulations. The greatest reductions in persistence occurred in scenarios simulated with impacts from drought in combination with fire or anthropogenically-mediated losses of hydrologic flows. Optimal actions to improve persistence were to prevent distant and smaller marshes from acting as sinks through strategic creation of megamarshes that act as sources of voles and stepping-stones. This research reinforces that management resources expended without guidance from empirically-based modeling can actually harm species’ persistence. This metapopulation-PVA tool could easily be implemented for other patchily-distributed endangered species and allow managers to maximize scarce resources to improve the likelihood of endangered species persistence.