Marco Ginepro

Vitality and genetic fidelity of white-rot fungi mycelia following different methods of preservation.

Basidiomycetes present specific problems with regard to their preservation, because most of them do not form resistant propagules in culture but exist only as mycelium. Usually these fungi can only be preserved by serial transfer on agar (labour-intensive procedures that can increase the danger of variation or loss of physiological or morphological features), or cryopreserved in liquid nitrogen (expensive). Cryopreservation at -80 degrees C and lyophilisation could be good alternatives. In this work we set up and tested six protocols of cryopreservation at -80 degrees C, and 12 protocols of lyophilisation on 15 isolates of white-rot fungi (WRF) belonging to 10 species. The tested protocols were mainly characterized by the use of different growth media, protectants, time and number of perfusion with protectants and finally by the typology and origin of the samples to be cryopreserved (mycelium/agar plug, whole colony) or to lyophilise (mycelium/agar plug, mycelium fragment, whole colony). Cryopreservation and lyophilisation outcomes were checked, at morphological (macro- and microscopic features), physiological (growth rate and laccase, Mn-independent and Mn-dependent peroxidases activities) and genetic level (Amplified Fragment Length Polymorphisms analysis - AFLP). Vitality of all fungi was successfully preserved by all cryopreservation protocols at -80 degrees C, and by two lyophilisation methods. Our results showed that cryopreservation at -80 degrees C did not produce morphological changes in any isolate, while two isolates were affected by lyophilisation. None of the physiological features were lost, even though growth rate and enzyme activities were somehow influenced by all preservation methods. AFLP analysis showed that only the two isolates that varied in their morphology after lyophilisation produced a different DNA fingerprint pattern in comparison with that obtained before lyophilisation. These findings provide evidence that cryopreservation at -80 degrees C and lyophilisation are suitable alternatives to liquid nitrogen cryopreservation for preservation of some WRF strains.

Benefits for agriculture and the environment from urban waste

Soluble bio-based substances (SBO) that have been isolated from urban biowaste have recently been reported to enhance plant leaf chlorophyll content and growth. The same SBO have also been shown to enhance the photochemical degradation of organic pollutants in industrial effluent. These findings suggest that SBO may promote either C fixation or mineralization, according to operating conditions. The present work aims to investigate SBO performance, as a function of source material. Thus, three materials have been sampled from a municipal waste treatment plant: (i) the digestate of the anaerobic fermentation of a humid organic fraction, (ii) a whole vegetable compost made from gardening residues and (iii) compost made from a mixture of digestate, gardening residues and sewage sludge. These materials were hydrolyzed at pH13 and 60°C to yield SBO that display different chemical compositions. These products were applied to soil at 30, 145 and 500 kg ha(-1) doses for tomato cultivation. Soil and plant leaf chemical composition, plant growth, leaf chlorophyll content and CO2 exchange rate as well as fruit quality and production rate were measured. Although it did not affect the soils chemical composition, SBO were found to significantly increase plant photosynthetic activity, growth and productivity up to the maximum value achieved at 145 kg ha(-1). The effects were analyzed as a function of SBO chemical composition and applied dose. The results of this work, compared with those of previous works, indicate that urban biowaste, if properly exploited, may furnish conjugate economic and environmental benefits, within a friendly sustainable ecosystem.

A study on the reproducibility of Tessier's extractions in a fluvial sediment and a comparison between different dissolution procedures in a reference material

Abstract The reproducibility of Tessiers extractions and the total content of cadmium, chromium, copper, iron, manganese, lead, zinc and calcium in a river sediment have been evaluated. The metals were determined with AAS in flame and in graphite furnaces. The accuracy of the dissolution procedures was evaluated using a reference material (RM) BCR 145; none of the methods applied proved optimal for all the metals determined. The concentrations of metals extracted by the various reagents were characterized by good reproducibility on species bonded to the carbonates, to iron and manganese oxides and in the residual; precision was lower in the other cases. The sequential extractions also showed a satisfactory mass balance.

Processed vs. non-processed biowastes for agriculture: effects of post-harvest tomato plants and biochar on radish growth, chlorophyll content and protein production.

The aim of this work was to address the issue of processed vs. non-processed biowastes for agriculture, by comparing materials widely differing for the amount of process energy consumption. Thus, residual post harvest tomato plants (TP), the TP hydrolysates obtained at pH 13 and 60 °C, and two known biochar products obtained by 650 °C pyrolysis were prepared. All products were characterized and used in a cultivation of radish plants. The chemical composition and molecular nature of the materials was investigated by solid state 13C NMR spectrometry, elemental analysis and potentiometric titration. The plants were analysed for growth and content of chlorophyll, carotenoids and soluble proteins. The results show that the TP and the alkaline hydrolysates contain lignin, hemicellulose, protein, peptide and/or amino acids moieties, and several mineral elements. The biochar samples contain also similar mineral elements, but the organic fraction is characterized mainly by fused aromatic rings. All materials had a positive effect on radish growth, mainly on the diameter of roots. The best performances in terms of plant growth were given by miscanthus originated biochar and TP. The most significant effect was the enhancement of soluble protein content in the plants treated with the lowest energy consumption non processed TP. The significance of these findings for agriculture and the environment is discussed.

Salt effects on the protonation of polymethacrylate and Na+, K+, Ca2+ complex formation

Abstract The protonation of polymethacrylic acid (W∼230 000) has been studied potentiometrically [(H+)-glass electrode] in different aqueous salt solutions (NaCl, KCl, Et4NI; 0≤I≤0.5 mol dm−3) at 25°C. Measurements have been performed also for polymethacrylic acid (PMA)–CaCl2 mixtures (PMA/Ca2+=0.5–4) at I=0.1 mol dm−3 (NaCl). The potentiometric data have been analyzed first by using the three-parameters model of Hogfeldt for apparent protonation constants, and then differences in different media have been interpreted in terms of complex formation. Formation constants are reported for Na+, K+ and Ca2+ complexes.

Digestion Procedures for the Elemental Analysis of Wood by Inductively Coupled Plasma Optical Emission Spectrometry

ABSTRACT Various digestion methods are available for the elemental analysis of wood biomass. Microwave digestion is the current European standard method for the determination of major and minor elements in solid biofuels. Because of the small masses (≤0.5 g) employed in microwave digestion, the resulting analyte concentrations in solution are low, and inductively coupled plasma mass spectrometry (ICP-MS) may be necessary to provide the required low limits of detection. However, higher concentrations are required for inductively coupled plasma optical emission spectrometry (ICP-OES) compared to ICP-MS. In this work, wet, dry, ash fusion, and microwave digestion were employed to determine Al, Ca, Fe, Mg, P, K, Si, Na, S, Cd, Co, Cr, Cu, Mn, Ni, Pb, and Zn in wood biomass by ICP-OES. Two certified reference materials (BCR 100, beech leaves and BCR 62, olive leaves) were used in this study. The concentrations of Cd, Co, Cr, Cu, Ni, and Pb were below the limits of quantification for ICP-OES. Wet digestion was suitable for the elemental analysis of wood biomass by ICP-OES with the exceptions of Al and Si. Dry digestion was accurate for some elements and ash fusion was the most accurate for the determination of silicon.

Plant response to biowaste soluble hydrolysates in hibiscus grown under limiting nutrient availability

ABSTRACT Biostimulants are substances promoting plant growth, quality and stress resistance. The present work aimed to investigate whether soluble hydrolysates from biowaste performed as biostimulants. Hibiscus (Hibiscus moscheutos L. subsp. palustris) plants were subjected to four treatments: standard fertilization, low fertilization (LF), and LF with added soluble digestate or soluble compost. Plant performance indicators were biomass accumulation, biometric parameters, leaf gaseous exchanges and elemental composition, and nitrogen (N)-use efficiency. LF negatively affected most of the investigated parameters. However, plants treated with biowaste-derived products performed better than untreated low-fertilized plants (+21 to 145% for biomass accumulation and biometric parameters, and carbon assimilation rate) and for many parameters reached values comparable to those showed by standard-fertilized plants or even higher, as in the case of Potassium (K), Silicon (Si), and Molybdenum (Mo) leaf content, and N use efficiency. Therefore, the tested soluble hydrolysates demonstrated to have biostimulant properties in hibiscus grown under nutritional stress.

Chemical Modeling of Acid-Base Properties of Soluble Biopolymers Derived from Municipal Waste Treatment Materials

This work reports a study of the proton-binding capacity of biopolymers obtained from different materials supplied by a municipal biowaste treatment plant located in Northern Italy. One material was the anaerobic fermentation digestate of the urban wastes organic humid fraction. The others were the compost of home and public gardening residues and the compost of the mix of the above residues, digestate and sewage sludge. These materials were hydrolyzed under alkaline conditions to yield the biopolymers by saponification. The biopolymers were characterized by 13C NMR spectroscopy, elemental analysis and potentiometric titration. The titration data were elaborated to attain chemical models for interpretation of the proton-binding capacity of the biopolymers obtaining the acidic sites concentrations and their protonation constants. The results obtained with the models and by NMR spectroscopy were elaborated together in order to better characterize the nature of the macromolecules. The chemical nature of the biopolymers was found dependent upon the nature of the sourcing materials.

Air pollution deposition on a roadside vegetation barrier in a Mediterranean environment: Combined effect of evergreen shrub species and planting density

Leaf deposition of PM10-100, PM2.5-10, PM0.2-2.5 and of 21 elements was investigated in a roadside vegetation barrier formed by i) two evergreen shrub species (Photinia × fraseri, Viburnum lucidum), with ii) two planting densities (0.5, 1.0 plant m-2), at iii) three distances from the road (2.0, 5.5, 9.0 m), at iv) two heights from the ground (1.5, 3.0 m), and on v) three dates (Aug, Sep, Oct). The presence of black and brown on-leaf PM10-100 and their element composition were detected by microscopy and image analysis. Pollutant deposition was also measured using passive samplers at five distances from the road (2.0, 5.5, 9.0, 12.5, 19.5 m) in the area of the barrier and in an adjacent lawn area. V. lucidum had more PM2.5-10 and PM0.2-2.5 on leaves than P. × fraseri, while most elements were higher in P. × fraseri. Most pollutants decreased at increasing distances from the road and were higher at 1.5 m from the ground compared to 3.0 m. Higher planting density in P. × fraseri enhanced the deposition of PM10-100 and PM2.5-10, while in V. lucidum, the planting density did not affect the depositions. Black PM10-100 decreased a long distance from the road and was entirely composed of carbon and oxygen, which was thus identified as black carbon from fuel combustion. The vegetation barrier had a higher deposition of most PM fractions at 5.5-12.5 m, while in the lawn area, depositions did not change. At 19.5 m, the PM10-100 was 32% lower behind the barrier than in the lawn area. In conclusion, the vegetation barrier changed the deposition dynamics of pollutants compared to the lawn area. These results strengthen the role of vegetation barriers and shrub species against air pollution and may offer interesting insights for the use of new road green infrastructures to improve air quality.