Short description
The experimental activities carried out at the BioSoundEcology Lab in Messina are focused on the analysis of biological underwater acoustic sources and the study of the ecological dynamics of marine vertebrates and invertebrates in polar habitats. In addition, the impacts of anthropogenic acoustic sources on the ecology, physiology and behaviour of marine organisms are assessed. In particular, through the study of the sounds generated by animals, the biological and ecological aspects are investigated. The analysis of the noise from human activities provides useful elements for understanding the effects on animals through the study of their behavioural and physiological responses.

Matrices of interest
The activities are mostly ascribable to marine habitats.

Study techniques
At the BioSoundEcology Lab, acoustic data acquisition in field/controlled environment and assessment of animal responses to environmental noise disturbance are carried out, in particular:
- Acoustic data analysis;
- Estimation of underwater noise and environmental acoustic components;
- Evaluation of ecological dynamics through the analysis of the vocalizations of marine animals;
-Analysis of the biochemical, physiological and behavioural responses of animals exposed to acoustic disturbance;
- Study of the acoustic ecology of marine mammals.

Instrumentation
The Lab is equipped with instrumentation for passive acoustic monitoring in the field (hydrophones and autonomous acoustic data acquisition systems) and for studies of the effects of noise on marine organisms in controlled environment (cameras, amplifier, acoustic transducer, software for acoustic and behavioral analysis).
For details: Dr. Francesco Filiciotto – francesco.filiciotto AT cnr.it

Short description
The research activities carried out in the BiogeM laboratory at the Messina headquarters are aimed at studying the marine and terrestrial biological processes that modulate and influence the chemical characteristics of the polar environment and the related cycles of matter and energy, also in relation to climate change.
Particular attention is paid to the evaluation of the role of microorganisms in the global carbon cycle and in the biogeochemical cycles of nutrients (nitrogen, phosphorus) of marine and lacustrine environments, through the study of both productive and degradative processes.
Among these, the activities involved in the decomposition of organic polymers, through microbial enzymatic activities, and the mineralization processes through microbial respiration.
The biogeochemical processes mediated by the microbial component are also being studied in the framework of the cryosphere, in order to understand the ecological significance of microbes in the permafrost, and their ability to maintain an active metabolism in extreme living conditions.
Together with the microbial processes linked to the biological pump and organic matter decomposition, a series of indirect biogeochemical parameters related to phytoplankton and bacterial biomass (Chlorophyll, ATP, lipopolysaccharides-LPS) are also determined.

Matrices of interest
The analyzed matrices are mostly ascribable to the hydrosphere (marine, river and lake waters, brines), soil, sediments, biofilm, cryosphere (permafrost, snow, sea and continental ice, intrapermafrost brines) and aquatic organisms.

Study techniques
At the BiogeM laboratory, measurements are carried out to determine the following parameters:
- Primary phytoplankton production;
- Microbial respiratory activity (consumed O₂; produced CO₂) (by ETS assay);
- Microbial enzymatic activities (leucin aminopeptidase, beta-glucosidase and alkaline phosphatase) (through fluorogenic substrates);
- Heterotrophic bacterial production;
- Content of total and size-fractionated (pico-, nano- and micro-phytoplanktonic) chlorophyll-a, pheopigments;
- microbial ATP in pico-, nano and microplankton fractions;
- Quantitative analysis of lipopolysaccharides (LPS) by the LAL chromogenic test.

Instrumentation
Spectrofluorimeter, Luminometer, Fluorometer, Spectrophotometer equipped with 96-well plate fluorescence reader, incubator, autoclave, balance, homogenizer, filtration systems, refrigerated centrifuge.
For information: Dr. Gabriella Caruso – gabriella.caruso AT cnr.it

Brief description
Experimental activities carried out at the EcoBiM LAB in Messina are addressed to the ecology and biotechnology of microorganisms, particularly prokaryotes, inhabiting both marine and continental polar habitats. The diversity of microorganisms, their response to environmental stress conditions (deriving from natural or anthropogenic forcing, such as climate change and chemical contamination), the astrobiological implications of life in extreme environments, and their evolution and adaptation in polar environments are among the ecological aspects investigated. EcoBiM researchers are also interested in the evaluation of the metabolic capabilities and biotechnological potentialities of cold-adapted microorganisms, by searching for biomolecules exploitable in industrial applications and bacteria able to degrade organic pollutants at low temperatures.

Matrices of interest
The analyzed matrices are mostly attributable to the hydrosphere (marine, river and lake waters, brines) and cryosphere (permafrost, snow, sea-ice and continental ice, intrapermafrost brines), but soils and sediments are also considered. The study of the interactions between microorganisms and biotic (for example, pelagic and benthic organisms) and abiotic (such as microbial communities colonizing polymeric materials, indicated with the term plastisphere) is of particular interest.

Applied experimental approaches
For the microbiological characterization of extreme environments, similarly to the analytical procedures commonly applied for the study of temperate areas, we use both culture-dependent and -independent (i.e. biomolecular and biochemical) approaches, including:
 - isolation and maintenance in pure culture of bacterial strains;
 - phenotypic (physiological, biochemical and morphological characteristics) and genotypic (analysis of the 16S rRNA sequences and search for functional genes) characterization of cultivable bacteria;
 - screening of bacteria for the production for useful biomolecules (including antibiotics, exopolysaccharides, biosurfactants);
 - evaluation of the metabolic capacities of the microbial communities through miniaturized assays;
 - characterization of the microbial communities by the hybridization in situ with oligonucleotide probes (CARD-FISH);
 - extraction of environmental DNA and RNA for metagenomics and metatrascriptomics studies;
 - preparation of microcosms enriched with organic and inorganic contaminants, and degradation tests.

Instrumentation
The EcoBiM LAB is equipped with basic instrumentation for environmental and applied microbiology (laminar flow cabinet, autoclave, incubators and thermostated baths, centrifuges, filtration systems, sonicators, spectrophotometers, fluorometers) and molecular biology equipment (thermocycler and equipment for electrophoresis).
For details: Dr. Angelina Lo Giudice - angelina.logiudice AT cnr.it

High Performance Liquid Chromatograph Agilent 1100 Series HPLC Systems

Brief description
The HPLC-MS/MS system allows to quantitatively determine polar organic compounds in several environmental and vegetable matrices and biota samples. It is commonly used to investigate specific markers of sources or environmental processes in samples collected in polar regions. It is used to determine levoglucosan, key tracer of biomass burning, in ice core samples to provide historical profiles of fire regimes in paleoclimatic studies. Several water soluble organic compounds (free and combined amino acids, phenolic compounds) are determined using HPLC-MS/MS in aerosol samples to define chemical composition of atmosphere in urban and polar samples. Polar pesticide or toxins are commonly determined with HPLC-MS/MS in fresh water, sea water, biota or vegetable samples.

Instrument
High Performance Liquid Chromatograph Agilent 1100 Series HPLC Systems (Waldbronn, Germany) with a binary pump, vacuum degasser, autosampler and thermostated column compartment coupled with an API 4000 Triple Quadrupole Mass Spectrometer (Applied Biosystem/MSD SCIEX, Concord, Ontario, Canada) using a TurboV source.
Contact person: Dr. Roberta Zangrando - roberta.zangrando AT cnr.it - CNR-ISP Venice Headquarters

Matrix and type of measurement
Analysis of discrete samples of several matrices: ice, snow, atmospheric aerosol, lacustrine water, fresh water, sea water, sediment, vegetable and biota samples. Analysis of polar organic compounds such as for example anhydrosugars, amino acids, phenolic compounds, organic acids.

Agilent 1100 series HPLC system coupled with API 4000, High Performance Liquid Chromatograph coupled with tandem mass spectrometer (HPLC-MS/MS). (IMAGE)

Liquid chromatograph UHPLC mod. Dionex Ultimate 3000 Dual Pump RS
Brief description
This instrument is the key system to perform semicontinuous analysis of organic compounds in ice core samples.

Instrument
Liquid chromatograph UHPLC mod. Dionex Ultimate 3000 Dual Pump RS (Thermo ScientificTM) with vacuum degasser, column thermostat.
Contact person: Dr. Elena Barbaro - elena.barbaro AT cnr.it - CNR-ISP Venice Headquarters

Matrix and type of measurement
Semicontinuos analysis of ice cores. This system is coupled with continuous flow analysis (CFA).

Dionex Ultimate 3000 Dual Pump RS Thermo Scientific, Liquid chromatograph UHPLC Dual Pump (IMAGE)

Mercur Plus - Analytik Jena AG, cold vapor atomic fluorescence spectroscopy (CV-AFS).
Brief description
Cold vapor atomic fluorescence spectroscopy (CV-AFS) is an analytical technique used for the quantification of mercury at trace/ultra-trace levels. This technique is mainly used on “clean” aqueous matrices (eg ice, snow, and water) from remote and uncontaminated areas.
The sensitivity of the instrument is fully harnessed by using official methods such as USEPA1631 version E or UNI-EN 15853: 2010. The Hg present in the matrix is oxidized to Hg²⁺ with BrCl solution and then reduced to elemental mercury(Hg⁰) con SnCl₂. The Hg⁰ is stripped from the aqueous matrix using an inert carrier gas (argon) and successively transported to gold traps for the pre-concentration by amalgam formation. Following thermal desorption at T between 450-500 °C, the Hg⁰ is desorbed from the gold traps and is transported into a quartz cell.
Light from a mercury vapor lamp passes through the quartz cell that contains the sample mercury in a flow of argon carrier gas and excites all the mercury atoms which then emit a characteristic fluorescence radiation at 253.7 nm. The amount of light emitted by the mercury atoms in the sample is proportional to the amount of mercury passing through the quartz cell. The CV-AFS of the CNR-ISP is located inside a dedicated clean room.

Instrument
Mercur Plus - Analytik Jena AG, cold vapor atomic fluorescence spectroscopy (CV-AFS).
Contact person: Dr. Massimiliano Vardè - massimiliano.varde AT cnr.it - CNR-ISP Venice Headquarters

Matrix and type of measurement
Aqueous matrices: atmospheric deposition, snow, ice, drinking water, mineral water, natural water, seawater. Mercury, as total mercury, or as dissolved, filtered and unfiltered after sample pre-treatment.

Mercur Plus - Analytik Jena AG, cold vapor atomic fluorescence spectroscopy (CV-AFS). (IMAGE)

Gas chromatograph coupled with mass spectrometer (GC-MS)

Brief description
The GC-MS system allows the quantification of volatile apolar compounds in environmental matrices. In the environmental field GC-MS finds main application in the determination of persistent organic pollutants (POPs) such as PCBs, PBDEs, PAHs, pesticides, in environmental matrices both in urban and remote areas such as polar areas. As well as it used in the determination of personal care products such as fragrances that have been observed not only in urban areas but also in Antarctica.
In the ISP-CNR there are 3 GC-MS systems. Among these, the system equipped with a cryogenic trap allows the preconcentration of volatile compounds (allowing the quantification of volatile compounds difficult to analyze in GC even at very low levels). There is also a GC-MS system equipped with a pyrolyser that allows the analysis of non-volatile materials such as plastic materials.

Instrument

• GC-MS 7890A-5975C (Agilent) /
• GC-MS GC7890A+MS5975C (Agilent) with cryogenic trap (MARKES Int)
• GC-MS GC6890+MS5973 (Agilent) with Pyrolysis system Pyroprobe 5000 Series
Contact person: Dr. Elena Argiriadis - elena.argiriadis AT cnr.it - CNR-ISP Venice Headquarters

Matrix and type of measurement
Analysis of discrete samples of several matrices: ice, snow, atmospheric aerosol, lacustrine water, fresh water, sea water, sediment, vegetable and biota samples. Compounds analyzed: Apolar volatile compounds such as PCBs, PBDEs, PAHs, pesticids, fragrances, sterols.

GC-MS 7890A-5975C Agilent (IMAGE)