Environmental Laboratory

Lectures (3 credits): Measure units. Heavy metals: chemistry and toxicology. Environmental sampling, samples storage. Spectroscopy: Electromagnetic radiation, Adsorption and emission spectroscopy: principles. Ultraviolet and Visible spectroscopy: Lambert Beer law, instrumentation and applications. Mass spectroscopy: theory, instrumentation, quadrupole detector, ICP-MS. Nuclear Magnetic Resonance (NMR): theory, instrumentation, chemical shift, spectra of elements other that H and C. Fundamentals of chromatography: retention time, selectivity, efficiency. Gas chromatography and HPLC, instrumentation, use. Workshop (3 credits): Safety rules in a chemical laboratory. Sampling on the site under investigation. Sample extraction, separation by TLC and HPLC and quantification by spectrophotometric methods of plant pigments. Analysis of heavy metals and other elements by means of ICP-MS. Recording NMR spectra of different nuclei.

Lectures (3 credits): The microbiological sampling. Methods of analysis and monitoring of environmental microbial communities. Cell count methods on pollutants, enrichment cultures, microcosms. Environmental genomics: polymerase chain reaction (PCR), quantitative Real Time PCR, clone libraries, denaturing gradient gel electrophoresis (DGGE), Sanger sequencing, Illumina sequencing. Biodegradation of organic pollutants and genetics: monoaromatic hydrocarbons, polycyclic aromatic hydrocarbons (PAH), aliphatics. Bacterial resistance and metabolism of heavy metals and Arsenic. Rhizosphere and endosphere microbiology: plant growth promoting bacteria, pollutants rhizo-degradation, metal translocation in plants (phytoremediation and food safety). Bioremediation of contaminated sites. Microbial life in extreme environments: the acid mine drainage. Microbial electrochemical systems: electroactive bacteria, microbial fuel cells, chemicals production from waste biomass. Seminars complete the preparation of the student: private bioremediation companies, public bodies involved in environmental monitoring and managing (ARPA, CNR, park authority).
Practical classes (3 credits): Safety in microbiology laboratories. Sampling on the site under investigation. Preparation and conservation of samples for microbiological and molecular analysis. Total aerobic, anaerobic bacterial plate count, MPN count of pollutant degrading bacteria. Phenotypic characterization of bacterial isolates: resistance to heavy metals and arsenic, mineralization of organic compounds, plant growth promotion (siderophores, IAA, EPS, phosphate solubilisation, ACC deaminase, mobility). DNA extraction from environmental samples. Preparation of polymerase chain reaction (PCR) and quantitative PCR (qPCR): 16S rRNA and functional genes. Electrophoresis run on agarose gel. Analysis of nucleotide sequences. Data analysis and discussion. Writing the report.

Plant stress physiology: molecular and physiological aspects
Approaches for the identification of genetic traits conferring plant tolerance to abiotic stresses
Relationships between environment quality and food security and safety
Plant-soil chemical, biochemical and microbiological interactions
Plant biosensors and bioindicators for monitoring environmental quality
Phytoremediation of contaminated soils and waters: opportunity and limits
Experimental/laboratory activities
Instrumental analysis techniques for studying: a) plant physiological status (leaf temperature by thermal IR imaging, leaf fluorescence and fluorescence imaging, chlorophyll content sensors, leaf photosynthesis, multi- and hyperspectral sensors); b) soil and plant contents in inorganic and organic pollutants (UV-Vis spectrometry, GC-MS, LC-MS, ICP-MS; C/H/N/S analyzer); d) pollutant fluxes in the framework of soil/plant/air relationships (IRMS-stable isotope ratio spectrometry)
Molecular and biochemical approaches (proteomics, metabolomics, ionomics) for studying plant physiological status and plant efficiency in phytoremediation techniques
Development of field sampling plans for the assessment of the presence of pollutants in soils, waters and plant material
Each year, together with the students, a real study case is chosen and a specific research, including field and laboratory activities, is chosen. In the last years the following cases have been addressed to: a) natural As contamination of a mountain stream and related effects on downstream soils and vegetation; b) use of plants for the phytoextraction of heavy metals from a polluted agricultural soil; c) phytostabilization and/ phytoremediation of urban soil contaminated by hydrocarbons and heavy metals; d) accidental leakage of oils from a buried pipeline: i) effects on soil and plants (wilds and crops); ii) evaluation of the potential use of plants for stabilization/remediation of the pollution.