Riccio, Lorenzo (2025) Application of digital pathology to the study of canine testicular damage induced by environmental pollutants. [Tesi di dottorato]

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Abstract

The purpose of this thesis is to develop and apply a multidisciplinary methodological framework for identifying, quantifying, and interpreting testicular alterations in dogs as potential sentinels of environmental exposure. Rather than a single longitudinal experiment, the work comprises three complementary studies addressing different levels of biological evidence. In the first chapter, we investigated associations between environmental contamination and canine testicular health, focusing on dogs from highly polluted areas of the Campania region. Using chemical, morphological, and immunohistochemical analyses, we observed that cadmium, arsenic, and uranium were the most relevant contaminants in the examined population. Higher tissue concentrations of these metals were associated with more severe testicular alterations, including degenerative and neoplastic lesions, although causal inference cannot be established due to the cross-sectional design and lack of a low-exposure geographic control. In the second chapter, we examined molecular and cytological correlates of these histopathological patterns. Oxidative stress markers suggested that reactive oxygen species may mediate cellular degeneration and metabolic imbalance. Altered protamine profiles and sperm morphology indicated potential impairment of chromatin packaging and sperm integrity, consistent with subfertility risk in exposed animals. These findings provide mechanistic insight but remain exploratory and qualitative in nature. The third chapter developed a computational pathology workflow to enhance diagnostic consistency in canine testicular disease. A convolutional neural network trained on a large archival dataset achieved accurate discrimination between neoplastic and non-neoplastic tissues and among tumour subtypes, demonstrating the potential of digital pathology to support objective and reproducible diagnostic assessment. Collectively, these studies illustrate how chemical, molecular, and computational approaches can converge to elucidate the complex relationships between environmental exposure and testicular pathology. While each dataset addresses a distinct analytical dimension, their directional coherence supports the plausibility of environmentally mediated reproductive impairment in dogs. Further integrated, longitudinal studies using matched samples are required to confirm causative mechanisms and to strengthen the translational relevance for One-Health applications.

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