HISTOLOGICAL, MOLECULAR AND CHEMICAL EVALUATIONS OF DOG TESTICLES FROM HIGHLY POLLUTED AREAS IN CAMPANIA REGION

In Campania Region an area known as “the Land of Fires” is considered an open-air dump due to illegal toxic waste disposal, raising concerns about environmental and reproductive health. Considering that humans and dogs share the same environment, leading to exposure to the same pollutants such as Heavy Metals (HMs), this study aimed to investigate the correlation between HMs exposure and testicular diseases in dogs. Fifty puberal male stray dogs living in high polluted areas in CR were enrolled for the study. A total of 100 testicular samples were collected and anal- ysed by histological, molecular and chemical methods. H&E stain assessed baseline morphology while immunofluorescence (IF) allowed to evaluate the localization of Connexin-43, α-SMA, and Vimentin. Immunohistochemistry (IHC) and Western Blot (WB) analyses were used to evaluate 17-β-HSD, P450 aromatase, and PCNA. Heavy metals (Pb, Cd, Hg, As, Sn, and depleted U) were quantified by Inductively Coupled Plasma Mass Spectrometry. Based on histological evaluation, samples were classified into four groups: A) complete spermatogenesis, B) incomplete, C) absent and D) neoplasia. IF revealed altered localization of junctional and cytoskeletal proteins in groups B–D. IHC and WB showed signif- icant downregulation of 17-β-HSD and upregulation of P450 aro- matase in groups C-D while PCNA levels were unchanged. Chemical analysis revealed a dose-dependent accumulation of HMs, with highest concentrations in group D (91%) followed by C (36%), B (31%), and A (27%). All neoplastic testes (D) showed aberrant accumulation of depleted U. These findings suggest that environmental HMs exposure may alter testicular architecture and enzyme expression, contributing to spermatogenic failure and neo- plastic transformation in canine testicles.