Groundwater biodiversity patterns in the Lessinian Massif of northern Italy

1. The distribution patterns of stygobiotic invertebrates were examined with a stratified sampling design at 197 sites selected among four hydrogeographic basins in the Lessinian Massif (northern Italy). The sites were approximately evenly distributed among four hydrogeological zones: unsaturated and saturated zone of karstic aquifers, and hyporheic and saturated zone of porous aquifers. 2. Outlying Mean Index (OMI) analysis which assesses deviation of habitat conditions from reference conditions, was used to evaluate the importance of 14 selected environmental variables in shaping groundwater biodiversity patterns in the region (total of 89 stygobiotic species). The measured variables explained 80% of the variability in the data set. 3. Sampling sites were distributed along the environmental gradients defined by OMI analysis. Significant differences were detected between karstic and porous site, as well as among sites located in the four hydrogeological zones. Differences among the four hydrogeographic basins were not observed. 4. Ordination of stygobiotic species along the environmental gradients was best explained by historical variables (mainly Wu¨ rmian glaciation and age of the underlying geological formation), while variables related to hydrogeology (mainly pH, calcium concentration and habitat fragmentation) influenced species distributions in the hydrogeological zones. An Environmental Integrity Index and nitrate concentration were significantly correlated with altitude, but appeared not to play a significant role in determining stygobiotic biodiversity patterns at the regional scale. 5. Results of the OMI analysis were highly significant for all taxa, suggesting that stygobiotic species are sensitive to the environmental factors studied. Thirty-five species showed high habitat specialisation (OMI index > 10). These species were usually rare and endemic to the Lessinian Massif. Most of them were found in a single hydrogeological zone. 6. Quaternary glaciations appear not to have lowered stygobiotic species richness in the Lessinian Massif. This may be because of the marginal location of the region with respect to the Wu¨ rmian glacier limit and because of extensive networks of fractures in the vadose zone of the karst, which may have allowed stygobionts to move deep down in the aquifers to seek refuge during surface freezing and to recolonise ancestral habitats after the glaciers retreated.