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Nitrogen deposition (Ndep) is considered a significant threat to plant diversity in grassland ecosystems around the world. The evidence supporting this conclusion comes from both observational and experimental research, with “space-for-time” substitution surveys of pollutant gradients a significant portion of the former. However, estimates of regression coefficients for Ndep impacts on species richness, derived with a focus on causal inference, are hard to locate in the observational literature. Some influential observational studies have presented estimates from univariate models, overlooking the effects of omitted variable bias, and/or have used  P -value-based stepwise variable selection (PSVS) to infer impacts, a strategy known to be poorly suited to the accurate estimation of regression coefficients. Broad-scale spatial autocorrelation has also generally been unaccounted for. We re-examine two UK observational datasets that have previously been used to investigate the relationship between Ndep and plant species richness in acid grasslands, a much-researched habitat in this context. One of these studies ( Stevens et al., 2004 ,  Science , 303: 1876–1879) estimated a large negative impact of Ndep on richness through the use of PSVS; the other reported smaller impacts ( Maskell et al., 2010 ,  Global Change Biology , 16: 671–679), but did not explicitly report regression coefficients or partial effects, making the actual size of the estimated Ndep impact difficult to assess. We reanalyse both datasets using a spatial Bayesian linear model estimated using integrated nested Laplace approximation (INLA). Contrary to previous results, we found similar-sized estimates of the Ndep impact on plant richness between studies, both with and without bryophytes, albeit with some disagreement over the most likely direction of this effect. Our analyses suggest that some previous estimates of Ndep impacts on richness from space-for-time substitution studies are likely to have been over-estimated, and that the evidence from observational studies could be fragile when confronted with alternative model specifications, although further work is required to investigate potentially nonlinear responses. Given the growing literature on the use of observational data to estimate the impacts of pollutants on biodiversity, we suggest that a greater focus on clearly reporting important outcomes with associated uncertainty, the use of techniques to account for spatial autocorrelation, and a clearer focus on the aims of a study, whether explanatory or predictive, are all required.

peerj

Reassessing the observational evidence for nitrogen deposition impacts in acid grassland: spatial Bayesian linear models indicate small and ambiguous effects on species richness