The set of living beings, animals and plants that inhabit our planet in their wide variety, as the result of long and complex evolutionary processes, represent the diversity of life on Earth, defined by the term biodiversity. During the twentieth century, the world has experienced an unprecedented technological and industrial explosion, that has had several negative consequences, including an increasing risk due to the effects that some compound and industrial discharges, means of threats to biodiversity (Massa, 2005; Agapow et al., 2004). Environmental degradation, the introduction of non-native species, over-exploitation of species, climate change, the presence of environmental contaminants, and the use of fertilizers, pesticides and often unpleasant herbicides exert toxic effects on the endocrine system and have, each, a prominent place among the risk factors for reproductive health and therefore for the maintenance of a specie (Hoffmann et al., 2010; Stork, 2010). Scientific research, lately, basically it’s oriented at the study of the toxic effects that many compounds have on molecules, cells, tissues, organisms and populations belonging to different classes of vertebrates with the aim to identifying, evaluating the damage and even more to predict the risk of species extinction (Mace et al., 2012). The following study, starting from biosensors potentially at risk of extinction species, such as Amphibians belonging to the genus Pelophylax, present along one of the most polluted river in Europe, develops and validates, with non-invasive biomarkers, skin and testis by gonadectomy, an innovative approach in environmental diagnostics for their preservation and the subsequent evaluation of the state of health of watercourses. In the first part of this research was detected and monitored the state of the health of the Plain of the Sarno river on the base of previous studies, by a synergic analysis of physical, chemical, biological and eco-toxicological components, which permitted to assess the ecological vulnerability of the river ecosystem, evidenced by the actual subletal stress on the organisms present in the Plain, due to the concentration of pollutants deriving from human and industrial activities. The study of the area allowed to define two favorite site, with different pollution degree. In the sites was monitored the presence of different frogs belonging to the genus Pelophylax, which were, in first instance, morphologically identified by classic taxonomic keys; since the difficult to specifically identify some specimens each other, due to the phenomenon of hybridization, only specimens of dubious belonging were analyzed by molecular approach, using the barcoding method. This even permitted to widen the genetic database for the specie Pelophylax bergeri, chosen as biosentinel specie. Furthermore, the evaluation of the oxidative and genotoxic damage on skin biopsy and gonadectomy, was carried out to the assessment of innovative non-invasive techniques. The second part of this research was devoted to develop an effective method for an eco-monitoring based on the analysis of ROS species in frog tissues, by an optimized spin-trapping EPR protocol. Furthermore, the genotoxic damage was detected by mobility shift, by monitoring the fragmentation degree of DNA, extracted by skin biopsy and gonadectomized testis. It’s interesting to note that, while the skin resulted to be damaged in both sites of Sarno River, testis seem to have developed a repair mechanism. This carried out at the study of enzymes involved in repair systems. After characterization and purification of the enzyme PARP-2, which is known to be responsible for the repair of DNA damage caused by heavy metals, the attention was paid to the testis. So, thanks to the examination of repair to DNA during spermatogenesis we realized the PARP, implicated in genome surveillance and protection, might represent in frog spermatogenesis an appealing tool for genotoxic risk assessment, useful to define a warning alarm for its survival.

THE SARNO RIVER AND THE BIO-PRESERVATION OF SPECIES OF THE GENUS Pelophylax / Trocchia, Samantha. - (2014).

THE SARNO RIVER AND THE BIO-PRESERVATION OF SPECIES OF THE GENUS Pelophylax

TROCCHIA, SAMANTHA
2014

Abstract

The set of living beings, animals and plants that inhabit our planet in their wide variety, as the result of long and complex evolutionary processes, represent the diversity of life on Earth, defined by the term biodiversity. During the twentieth century, the world has experienced an unprecedented technological and industrial explosion, that has had several negative consequences, including an increasing risk due to the effects that some compound and industrial discharges, means of threats to biodiversity (Massa, 2005; Agapow et al., 2004). Environmental degradation, the introduction of non-native species, over-exploitation of species, climate change, the presence of environmental contaminants, and the use of fertilizers, pesticides and often unpleasant herbicides exert toxic effects on the endocrine system and have, each, a prominent place among the risk factors for reproductive health and therefore for the maintenance of a specie (Hoffmann et al., 2010; Stork, 2010). Scientific research, lately, basically it’s oriented at the study of the toxic effects that many compounds have on molecules, cells, tissues, organisms and populations belonging to different classes of vertebrates with the aim to identifying, evaluating the damage and even more to predict the risk of species extinction (Mace et al., 2012). The following study, starting from biosensors potentially at risk of extinction species, such as Amphibians belonging to the genus Pelophylax, present along one of the most polluted river in Europe, develops and validates, with non-invasive biomarkers, skin and testis by gonadectomy, an innovative approach in environmental diagnostics for their preservation and the subsequent evaluation of the state of health of watercourses. In the first part of this research was detected and monitored the state of the health of the Plain of the Sarno river on the base of previous studies, by a synergic analysis of physical, chemical, biological and eco-toxicological components, which permitted to assess the ecological vulnerability of the river ecosystem, evidenced by the actual subletal stress on the organisms present in the Plain, due to the concentration of pollutants deriving from human and industrial activities. The study of the area allowed to define two favorite site, with different pollution degree. In the sites was monitored the presence of different frogs belonging to the genus Pelophylax, which were, in first instance, morphologically identified by classic taxonomic keys; since the difficult to specifically identify some specimens each other, due to the phenomenon of hybridization, only specimens of dubious belonging were analyzed by molecular approach, using the barcoding method. This even permitted to widen the genetic database for the specie Pelophylax bergeri, chosen as biosentinel specie. Furthermore, the evaluation of the oxidative and genotoxic damage on skin biopsy and gonadectomy, was carried out to the assessment of innovative non-invasive techniques. The second part of this research was devoted to develop an effective method for an eco-monitoring based on the analysis of ROS species in frog tissues, by an optimized spin-trapping EPR protocol. Furthermore, the genotoxic damage was detected by mobility shift, by monitoring the fragmentation degree of DNA, extracted by skin biopsy and gonadectomized testis. It’s interesting to note that, while the skin resulted to be damaged in both sites of Sarno River, testis seem to have developed a repair mechanism. This carried out at the study of enzymes involved in repair systems. After characterization and purification of the enzyme PARP-2, which is known to be responsible for the repair of DNA damage caused by heavy metals, the attention was paid to the testis. So, thanks to the examination of repair to DNA during spermatogenesis we realized the PARP, implicated in genome surveillance and protection, might represent in frog spermatogenesis an appealing tool for genotoxic risk assessment, useful to define a warning alarm for its survival.
2014
THE SARNO RIVER AND THE BIO-PRESERVATION OF SPECIES OF THE GENUS Pelophylax / Trocchia, Samantha. - (2014).
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11588/593931
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