Penconazole is one of the most widely used fungicides all over the world, and since it spreads to large environments, its toxic effects on non-target organisms are of great concern. The toxic effects of penconazole on crayfish (Astacus leptodactylus), which is a bioindicator in freshwater ecosystems and consumed economically, are not known. Therefore, in this study, the purpose was to contribute to the literature on the potential harmful effects of penconazole on a non-target species, Astacus leptodactylus. For this aim, the acute toxicity (96 h) of penconazole was examined. The 96-h LC₅₀ value of penconazole was detected as 18.7 mg L⁻¹. Four concentrations of penconazole (18.7 mg L⁻¹, 9.35 mg L⁻¹, 4.68 mg L⁻¹, 2.34 mg L⁻¹) were applied to crayfish for 96 h. The results showed that penconazole had destructive effects on esterase mechanisms by inhibiting acetylcholinesterase (AChE) and carboxylesterase (CaE) activities. Significant increases were observed in all antioxidant parameters (superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), reduced glutathione (GSH), malondialdehyde (MDA)) in all doses except the lowest concentration (2.34 mg L⁻¹). All adenosine triphosphatase (ATPase) activities (Na⁺/K⁺-ATPase, Mg²⁺-ATPase, Ca²⁺-ATPase, total ATPase) had significant dose-related inhibition in both gill and muscle tissues. In summary, our findings show that acute penconazole administration to crayfish causes significant toxic effects on esterase, antioxidative parameters, and metabolic enzymes.

Freshwater crayfish are bioindicators of environmental pollution, often used for the assessment of heavy metal (HM) presence in the tissues, a time-consuming and expensive task. In this study, we propose the use of the vibrational spectroscopy to detect in a fast, non-destructive and sensitive way the presence of HM in the cephalothorax exoskeleton of the freshwater crayfish. Incorporation of HM into the cephalothorax exoskeleton was investigated under controlled laboratory conditions. In particular, the cephalothorax exoskeleton of five crayfish species (Astacus leptodactylus, Procambarus clarkii, Austropotamobius pallipes, Faxonius limosus, and Pacifastacus leniusculus) was analyzed by attenuated total reflection–Fourier transformed infrared (ATR–FTIR) spectroscopy in the presence or absence of cadmium (Cd), chromium (Cr), lead (Pb), nickel (Ni), and zinc (Zn) up to 4 weeks at various concentrations (0.01, 0.1, 1, 10, ppm). The ATR–FTIR profile of the crayfish cephalothorax exoskeleton was compatible with the presence of amorphous calcium carbonate, chitin, and proteins. The incubation with the HM revealed two main modifications: the shift of the peak from 859 to 872 cm⁻¹ and the appearance of a peak at 712 cm⁻¹. Both are ascribable to the HM interaction with calcium carbonate. The absorbance of both peaks increased along with the time of incubation, and the HM concentration. We conclude that ATR–FTIR analysis can be a useful, quick, and cost-sensitive tool to detect HM presence in the crayfish cephalothorax exoskeleton. However, it has to be regarded as a non-specific analytical technique for assessing HM contamination, since it is unable to discriminate between different HM.