Chianese, Teresa (2025) Evaluation of the anthropogenic impact on animal reproduction using in vivo and in vitro models. [Tesi di dottorato]

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Abstract

Human activities are profoundly altering all ecosystems. The overexploitation of land and water resources, combined with pollution, generates complex and often difficult-to-measure adverse synergistic effects. The contaminants responsible for this phenomenon are numerous, ranging from heavy metals to pesticides, herbicides, pharmaceuticals, and even micro- and nanoplastics. One of the emerging pollutants is plastic, the most widespread material in the world after steel and concrete. Its history begins in 1861, when Alexander Parkes, working on cellulose nitrate, patented the first semi-synthetic plastic material, called Parkesine (M. Pivato and Donzelli, 2012). Despite the central role of plastics in modern society, their environmental impact can be dramatic if not properly managed. Their resistance to degradation makes them persistent in the environment, transforming them from an innovative resource into a major source of pollution. In addition to plastics, pesticides also represent an emerging category of pollutants, as agriculture, one of the oldest human activities, is also affected by pressures related to population growth and increasing food demand. The intensive use of pesticides, while contributing to crop protection and increased productivity, raises growing concerns for environmental and human health. A case in point is glyphosate [N-(phosphonomethyl)glycine], a non-selective systemic herbicide absorbed through the leaves and transported throughout the plant via the phloem. This compound is known for its persistence in soil and its ability to contaminate groundwater, leading to significant impacts on aquatic ecosystems (Mertens et al., 2018). Numerous studies have highlighted correlations between exposure to glyphosate and the onset of neoplastic diseases and autoimmune disorders. Reproductive toxicity is considered the main cause of biodiversity decline. The production of high-quality gametes is a prerequisite for proper fertility, which must be followed by adequate embryonic development. My doctoral project fits into this context, aiming to investigate the effects of environmental pollutants, particularly microplastics and glyphosate, on animal reproduction, using integrated vivo and in vitro approaches. Three main models were used: Mytilus galloprovincialis and Podarcis siculus for in vivo studies on the effects of microplastics, heavy metals and the herbicide glyphosate, and the human PNT1a cell line for in vitro studies on the effects of glyphosate. Together, these results showed how different classes of environmental pollutants, despite acting through distinct mechanisms, including mechanical interference in the case of microplastics, converge toward the same outcome: compromised reproductive capacity. This convergence highlights the universal threat posed by anthropogenic pollution, which transcends ecological boundaries and species barriers, indiscriminately affecting marine and terrestrial organisms. Considering that the simultaneous presence of multiple contaminants is now common, this information raises concerns about the additive toxic effects of substances, even those that are chemically different. On the other hand, it enables the scientific community to identify the mechanisms underlying cellular toxicity, regardless of the substance, and to implement strategies for restoring animal welfare. The results of these three years of research highlight the harmful effects of some major classes of environmental pollutants, such as microplastics, heavy metals and glyphosate on animal reproductive organs, using a multidisciplinary approach. In particular, in mussels, microplastics and heavy metals caused cell damage, inflammation, and metabolic imbalances, with a potential reduction in gametogenesis and reproductive success. In P. siculus, glyphosate acted as a potent endocrine disruptor, altering the structure and function of the gonads and modifying the expression of oestrogen receptors, probably due to its oestrogen-like activity. In vitro experiments on human prostate cells (PNT1A) confirmed this effect, showing that glyphosate behaves as a xenoestrogen, inducing oxidative stress, apoptosis and bioenergetic imbalances. Overall, the results indicate that different environmental pollutants, while acting through distinct mechanisms, converge on the same outcome: the impairment of reproductive capacity, thus threatening biodiversity and ecosystem health

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