Mauro Mecozzi

Baseline Trace Metals in Seagrass, Algae, and Mollusks in a Southern Tyrrhenian Ecosystem (Linosa Island, Sicily)

Trace elements were analyzed in organisms collected at five sampling stations along coastal areas of Linosa Island, Sicily (southern Tyrrhenian Sea, Italy). Concentrations of Cd, Cr, Cu, Pb, and Zn were measured in Posidonia oceanica L. Delile tissues, the two brown algae Padina pavonica (L.) Thivy and Cystoseira sp., and the two gastropod mollusks Monodonta turbinata Born and Patella caerulea L. Seawater samples were also collected at each site to assess soluble metal concentrations and to gain relevant information on their bioaccumulation ability. Data were processed by multivariate techniques, such as principal component analysis (PCA) and linear discriminant analysis on PCA factors. The scoreplots obtained showed that the pollutant distribution is more significantly correlated with species than with sites. For seaweeds, P. oceanica was associated with higher Cd, Cu, and Zn levels; Padina species had higher Cr concentrations, and Cystoseira had higher Pb levels. For mollusks, Monodonta species had high concentrations of Cu and Cr and Patella species were associated with Cd. Some general metal bioaccumulation patterns are described but no one sampling site was more contaminated than the others. The hypothesis of Linosa island serving as a reference ecosystem for baseline trace metal levels in southern Tyrrhenian areas is indeed supported by the statistical comparison among other southern Tyrrhenian ecosystems performed with Kruskall Wallis and Mann-Whitney tests. For P. oceanica leaves, P. pavonica, M. turbinata, and P. caerulea, this study confirms their usefulness as possible cosmopolitan biomonitors of trace metals in marine Mediterranean areas.

Characterization of mucilage aggregates in Adriatic and Tyrrhenian Sea: structure similarities between mucilage samples and the insoluble fractions of marine humic substance

The appearance of gelatinous aggregates called mucilages causes serious damages to tourism and fishery industries of the Adriatic Sea. So, many studies have been planned and some of them are still in progress to clarify the origin and causes of the phenomenon. The scientific research has showed that mucilages are produced by several marine organisms when peculiar climatic and trophic conditions occur. Moreover, as far as the mucilage composition is concerned, although it is well known that polysaccharides give a high contribution, knowledge of the structural characteristics of mucilages and their relationship with the natural organic matter of the marine environment has not been clarified yet. In this paper a study on the characterization of the marine mucilage samples collected in the Adriatic and Tyrrhenian Seas is described. The study was performed by spectroscopic (infrared and colorimetric) techniques, and elemental analysis. The results showed that mucilage samples have chemical and structural similarities with the insoluble fraction of the marine humic substance (humin). According to experimental evidences it is possible to establish the relationship between mucilages and the dissolved organic matter (DOM) in the marine environment in order to identify the most likely pathways of mucilage formation.

An ultrasound assisted extraction of the available humic substance from marine sediments

In this paper an ultrasound assisted procedure for the extraction of the bio-available fraction of humic substance in marine sediments is described. The proposed method is based on a preliminary 24 h 8 M HCl treatment already proposed in a previous paper [M. Mecozzi, E. Pietrantonio, M. Amici, Fres. Environ. Bull. 7 (1998) 605], followed by consecutive extractions by 0.5 M NaOH coupling to an ultrasonic treatment. The main advantages of the ultrasound procedure are the reduced times of extraction which take 30 min in contrast to the 24 h required by shaking method and the possibility to perform also the quantitative estimation of the extractable fraction of humic substance present in marine samples.

Molecular and structural characteristics in toxic algae cultures of Ostreopsis ovata and Ostreopsis spp. evidenced by FTIR and FTNIR spectroscopy.

In this article we investigated the compositional and structural characteristics of the principal biomolecules such as carbohydrates, proteins, lipids, nucleic acids and chlorophyll pigments present in biofilm cultures of Ostreopsis spp. and in batch cultures of Ostreopsis ovata. Our approach based on the use of infrared (FTIR) and near infrared (FTNIR) spectroscopy showed the marked differences existing between biofilm cultures and batch cultures. FTIR spectroscopy showed the higher contents of polysaccharides and chlorophyll pigments in O. ovata from batch cultures with respect to Ostreopsis spp. Second derivative FTIR spectroscopy showed different features concerning the secondary structure of proteins because in O. ovata samples the beta sheet and beta turn structures were observed whereas in Ostreopsis spp. samples the alpha helix structure was the most evident. FTNIR spectroscopy showed other structural differences observed existing between O. ovata and Ostreopsis spp. mainly related to hydrogen bond interactions determining more packed structures in the nucleus of O. ovata. In addition, the interpretation of FTIR and FTNIR spectral information was also supported by the application of two statistical methods, the independent component analysis (ICA) and the spectral cross correlation analysis (SCCA). ICA was used as spectral deconvolution technique to separate the effects of the interference bicarbonate ion from algal FTIR spectra so to verify the high similar qualitative composition of the three biofilm samples of Ostreopsis spp. At last, SCCA applied to FTIR and FTNIR spectra was useful to evidence some structural differences involving -CH and CH(2) groups of aliphatic chains in O. ovata and Ostreopsis spp. samples. Though preliminary, these results agree with some previous studies suggesting that the presence of different ecophysiological characteristics in O. ovata and Ostreopsis spp. depending on the parameters related to the condition growth.

The roles of carbohydrates, proteins and lipids in the process of aggregation of natural marine organic matter investigated by means of 2D correlation spectroscopy applied to infrared spectra.

In this paper the marine organic matter soluble in an alkaline medium called extractable humic substance (EHS), was extracted from three sediment samples of Tyrrhenian Sea and separated by precipitation at pH 2 in the two fractions of fulvic acids (FAs) and humic acids (HAs). FAs were further fractionated in seven sub-samples of different molecular weight (mw) by means of seven different ultrafiltration membranes operating in the range between mw<1 kDa and mw>100 kDa. Then the qualitative composition of each sample of fractionated FAs and HAs was studied by means of one-dimensional Fourier transform infrared spectroscopy in reflectance mode (FTIR-DRIFT) and by two-dimensional (2D) correlation spectroscopy both in wavelength-wavelength (WW) and in sample-sample (SS) mode. The application of 2D correlation WW spectroscopy allows to elucidate the different roles played by carbohydrates and proteins with respect to some lipid compounds such as fatty acids and ester fatty acids during the process of aggregate formations from mw approximately 1 kDa to higher size aggregates. In addition, 2D correlation WW spectroscopy allows to observe some peculiar interactions between carbohydrates and proteins in the formation of EHS aggregates, interactions which vary from a sample to another sample. The results of 2D correlation SS spectroscopy confirm the general evidences obtained by 2D WW spectroscopy and moreover, they also describe the formation of EHS aggregates as a complex process where evolutionary links and connectivity between aggregates of neighbour molecular size ranges are not evident. Two-dimensional correlation spectroscopy applied to FTIR spectroscopy shows to be a powerful tool for the investigation of the mechanisms involved in EHS aggregation because it supports the acquisition of structural information which sometimes can be hardly obtained by one-dimensional FTIR spectroscopy.

Ultrasound-assisted analysis of total carbohydrates in environmental and food samples

Analytical methods to determine total carbohydrates in environmental and food samples usually require a preliminary chemical hydrolytic procedure to convert polysaccharides into monosaccharides prior to detection by colorimetric or chromatographic techniques. In this paper, an alternative hydrolytic procedure is presented. The method for hydrolysis of polysaccharides is based on the application of ultrasound at room temperature. The advantages of the method proposed here are reduced time required for analysis (4-5 h) and the improvement of the analytical accuracy due to the absence of the most common reactions of oxidation. The proposed method was applied to the determination of total carbohydrates in several environmental samples (seawater and marine mucilage) and starchy food samples. Results for total carbohydrate obtained using the ultrasound procedure for hydrolysis of samples agreed with those found when applying other published procedures.

Determination of carbonate in marine solid samples by FTIR-ATR spectroscopy

A method for detecting carbonate in marine solid samples (sediments, corals) by Fourier transform infrared spectroscopy (FTIR) coupled to the total attenuated reflectance (ATR) technique is described. Compared to other techniques, the proposed method is not based on the measurement of CO2 evolved by combustion or acidification of the sample, but on the direct measurement of carbonate present in the sample. For this reason, the method by FTIR-ATR spectroscopy does not require any chemical pre-treatment. The proposed method allows determination of carbonate in the range 6-100% (w/w) as Na2CO3 and gives comparable results with the determination of inorganic carbon by elemental analysis.

Determination of Seawater Salinity by Ultraviolet Spectroscopic Measurements

A method for the determination of seawater salinity by ultraviolet (UV) spectroscopy is proposed. The effects of single salt concentrations and of salinity on UV absorption in the 190–250-nm range were investigated. These studies revealed that the absorption spectrum of a solution with a given salinity is due mainly, in order, to KBr >MgCl 2 >NaCl. The influence of the temperature and salt concentration on UV spectra was studied by using synthetic seawater samples with the salinities ranging from 1 to 50 parts per thousand (%). Results showed that, in the absence of interferences, the most sensitive and reliable conditions for measuring the salinity are at 212 nm and at temperatures in the range of 25–30 °C. Under these conditions this method shows quite linear calibration curves and allows us to perform salinity determinations in seawater solutions at concentrations as low as 4%. Moreover, it requires no sample pretreatment and offers a precision of 0.20%. The proposed method is very simple and rapid for laboratory and on-board analysis. Finally, the interference of organic matter, nitrite, and nitrate species with the salinity determinations was investigated. These studies show that organic matter does not interfere at concentrations of carbon lower than 1 mg/L and at 210 nm. Interferences due to NO− x species can be ignored if these species are dissolved in solution at concentrations lower than ≈0.2 mg/L and the analyses are carried out at wavelengths lower than 212 nm.

Improvement of an ultrasound assisted method for the analysis of total carbohydrate in environmental and food samples.

The determination of the carbohydrate content of environmental and food samples, performed by spectroscopic and chromatographic methods, needs a preliminary hydrolysis which converts polysaccharides into monosaccharides prior to the analytical measurement. In this paper, a modified procedure of the ultrasound-assisted method for the determination of total carbohydrates, published by the same authors in a previous paper, is presented and discussed. The modified procedure represents an improvement with respect to the previous one because it shortens the hydrolysis time and the colorimetric determination times through the use of two acid media (formic and acetic acid) that also allows quantitative recoveries of total carbohydrates from food and environmental samples.

Monitoring of marine mucilage formation in Italian seas investigated by infrared spectroscopy and independent component analysis

The aim of this study is to present and to discuss some characteristics of recalcitrant organic matter mechanism and formation. These aggregates called mucilages that are produced by the degradation reactions of several algae, have been investigated by infrared (FTIR) spectroscopy. FTIR spectra of macroaggregates produced by different algal samples have been daily collected in order to investigate the steps of aggregation. Afterwards, they have been elaborated by means of Independent Component Analysis (ICA). ICA investigation of FTIR spectra showed that the global aggregation process of marine mucilage always consisted of two different phases or independent components (ICs). One IC is related to the first degradation step of algal cells leading to the production of mono and oligosaccharides with aminoacids and oligopeptides. The second IC is related to the polymerization of oligosaccharides with aminoacids and oligopeptides and to their interaction with less polar compounds such as lipids thus producing supramolecular structures. The emerging mechanisms of anomalous size aggregates of organic matter match those of natural organic matter aggregation. The approach we suggest is to use synthetic mucilages which allows to monitor the macroaggregates formation because it can hardly be performed by means of natural marine macroaggregates.