Small heat shock proteins as relevant biomarkers for anthropogenic stressors in earthworms

Anthropogenic stressors in terrestrial ecosystems require focused research on adaptive responses in soil organisms such as Eisenia fetida, a model earthworm species. We analyzed the gene expression of five small heat shock proteins (sHSPs) in response to various stressors: heat stress (31 and 35 °C), desiccation (10 % and 20 % humidity), and chemical exposure (bisphenol A and endosulfan) under standard and elevated temperatures. Under moderate heat (31 °C), early upregulation of sHSP transcripts suggests their involvement in initial stress responses, possibly mitigating protein aggregation. At the higher temperature (35 °C), three sHSPs served as a defense against severe protein aggregation, a significant finding as previous studies identified only one activated heat shock protein (HSP70) in E. fetida under similar conditions. Desiccation stress at 10 % humidity activated more sHSPs than at 20 % humidity, and the expression profile at 10 % humidity closely resembled that observed under heat stress, suggesting overlapping adaptation pathways. Heat combined with chemical stress, particularly endosulfan, elevated sHSP transcription and underscored the potential of these proteins as biomarkers in multi-stressor environments. Monomeric sHSPs from E. fetida, which share homology with human sHSPs, showed the highest activity across all stressors, suggesting their key role in earthworm adaptation.