Giovanni Garau

The symbiotic requirements of different medicago spp. Suggest the evolution of sinorhizobium meliloti and S. medicae with hosts differentially adapted to soil pH

Nitrogen fixing rhizobia associated with the Medicago L. genus belong to two closely related species Sinorhizobium medicae and S. meliloti. To investigate the symbiotic requirements of different Medicago species for the two microsymbionts, 39 bacterial isolates from nodules of eleven Medicago species growing in their natural habitats in the Mediterranean basin plus six historical Australian commercial inocula were symbiotically characterized with Medicago hosts. The bacterial species allocation was first assigned on the basis of symbiotic proficiency with M. polymorpha. PCR primers specific for 16S rDNA were then designed to distinguish S. medicae and S. meliloti. PCR amplification results confirmed the species allocation acquired in the glasshouse. PCR fingerprints generated from ERIC, BOXA1R and nif-directed RPO1 primers revealed that the Mediterranean strains were genetically heterogenous. Moreover PCR fingerprints with ERIC and BOX primers showed that these repetitive DNA elements were specifically distributed and conserved in S. meliloti and S. medicae, clustering the strains into two divergent groups according to their species. Linking the Sinorhizobium species with the plant species of origin we have found that S. medicae was mostly associated with medics well adapted to moderately acid soils such as M. polymorpha, M. arabica and M. murex whereas S. meliloti was predominantly isolated from plants naturally growing on alkaline or neutral pH soils such as M. littoralis and M. tornata. Moreover in glasshouse experiments the S. medicae strains were able to induce well-developed nodules on M. murex whilst S. meliloti was not infective on this species. This feature provides a very distinguishing characteristic for S. medicae. Results from the symbiotic, genotypic and cultural characterization suggest that S. meliloti and S. medicae have adapted to different Medicago species according to the niches these medics usually occupy in their natural habitats.

Influence of selected lab cultures on the evolution of free amino acids, free fatty acids and Fiore Sardo cheese microflora during the ripening

Fiore Sardo Protected Denomination of Origin is a traditional Sardinian (Italy) hard cheese produced exclusively from whole raw ovine milk and coagulated with lamb rennet paste. Currently, Fiore Sardo is still produced by shepherds at the farmhouse level without the addition of any starter culture and the cheese-making process is characterized by significant waste. The first objective of the present work was to investigate the autochthonous microflora present in milk and Fiore Sardo cheese in order to select lactic acid bacterial (LAB) cultures with suitable cheese-making attributes and, possibly reduce the production waste. Secondly, the ability of selected cultures to guarantee cheese healthiness and quality was tested in experimental cheese-making trials. In this study, we show that the typical lactic microflora of raw ewes milk and Fiore Sardo cheese is mostly composed of mesophilic LAB such as Lactococcus lactis subsp. lactis, Lactobacillus plantarum and Lactobacillus casei subsp. casei. Moreover, strains belonging to the species were selected for cheese-making attributes and used in experimental cheese-making trials carried out in different farms producing Fiore Sardo. The evolution of the cheese microflora, free amino acids and free fatty acids during the ripening showed that the experimental cheeses were characterized by a balanced ratio of the chemical constituents, by a reduced number of spoilage microorganisms and, remarkably, by the absence of production waste that were significant for the control cheeses.

Copper(II) and lead(II) removal from aqueous solution by water treatment residues

In this study, we investigated the ability of Fe- and Al-based water treatment residues (Fe- and Al-WTR) to accumulate Pb(II) and Cu(II) at pH 4.5. The role of the inorganic and organic fractions of WTRs in metals sorption was also assessed. Sorption isotherms showed a higher sorption of Pb(II) by both WTRs with respect to Cu(II) (e.g. 0.105 and 0.089 mmol g(-1) of Pb(II) and Cu(II) respectively sorbed by Fe-WTR). Fe-WTR revealed a stronger sorbent for both metals than Al-WTR. The amount of Pb(II) and Cu(II) sorbed by Fe-WTR was about the 69% and 63% higher than that sorbed by the Al-WTR. The organic matter of Fe- and Al-WTR contributed to about 26% and 8.5% respectively in the sorption of both metals. The sequential extraction procedure showed that the greatest amount of metals sorbed by both WTRs were tightly bound and not extractable, and this was particularly apparent for Cu(II). The FT-IR spectra indicated the formation of inner-sphere complexes between the Fe(Al)-O nucleus and Pb(II) and Cu(II). Moreover, the FT-IR spectra also suggested that the humic fraction of WTRs interacted, through the carboxylate groups, with Cu(II) and Pb(II) by forming mainly monodentate and bidentate complexes, respectively.

Burkholderia rhynchosiae sp. nov. isolated from Rhynchosia ferulifolia root nodules

Two strains of Gram-stain-negative, rod-shaped bacteria were isolated from root nodules of the South African legume Rhynchosia ferulifolia and authenticated on this host. Based on phylogenetic analysis of the 16S rRNA gene, strains WSM3930 and WSM3937(T) belonged to the genus Burkholderia, with the highest degree of sequence similarity to Burkholderia terricola (98.84 %). Additionally, the housekeeping genes gyrB and recA were analysed since 16S rRNA gene sequences are highly similar between closely related species of the genus Burkholderia. The results obtained for both housekeeping genes, gyrB and recA, showed the highest degree of sequence similarity of the novel strains towards Burkholderia caledonica LMG 19076(T) (94.2 % and 94.5 %, respectively). Chemotaxonomic data, including fatty acid profiles and respiratory quinone data supported the assignment of strains WSM3930 and WSM3937(T) to the genus Burkholderia. DNA-DNA hybridizations, and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strains WSM3930 and WSM3937(T) from the most closely related species of the genus Burkholderia with validly published names. We conclude, therefore, that these strains represent a novel species for which the name Burkholderia rhynchosiae sp. nov. is proposed, with strain WSM3937(T) ( = LMG 27174(T) = HAMBI 3354(T)) as the type strain.

Sorption behavior of sulfamethazine on unamended and manure-amended soils and short-term impact on soil microbial community

In this study we investigated the sorption of sulfamethazine (SMZ) in two soils with different physico-chemical properties and the sulfonamide short-term influence on the microbial community structure and function. The presence of manure increased the SMZ sorption, the uppermost level being measured on soil with the lower pH and the higher manure content allowed by the Italian regulation. The sulfonamide desorption was hysteretic on both soils. SMZ addition to soil at the concentration of 53.6 μg/g had a significant short-term negative impact on readily culturable bacteria, potential metabolic activity (Biolog CLPP) and soil enzyme activity. Moreover, a shift of the culturable microbial populations towards a lower bacteria/fungi ratio was observed after SMZ addition. Despite the observed SMZ effects disappeared almost completely after 7 day, structural changes of microbial communities were still present in SMZ-treated soils. The results presented are remarkable since previous studies addressing the SMZ impact on soil microbial parameters failed to highlight any significant effect of the sulfonamide on microbial abundance and diversity.

Water treatment residues as accumulators of oxoanions in soil. Sorption of arsenate and phosphate anions from an aqueous solution

Here we report a survey addressed to determine, at different pH values (pH 4.0, 7.0 and 9.0), the ability of two different water treatment residues, a Fe-based (Fe-WTR) and an Al-based (Al-WTR), to accumulate arsenate and phosphate anions from an aqueous solution and to define the mechanism which regulate the sorption of these anions. Fe-WTR showed a greater As(V) and P(V) sorption capacity respect to Al-WTR at all the pH values investigated, in particular at pH 4.0. The greater capacity of the Fe-WTR to accumulate phosphate at pH 4.0 seems to be linked to the higher content of manganese ions compared to Al-WTR, which can give rise, with phosphate ions, to the formation of MnHPO4 precipitates. Sequential extraction of As(V)- or P(V)-WTRs suggested that the main mechanism governing the sorption of both two anions likely involve the formation of inner-sphere surface complexes [Fe/Al-O-As(P)]. Such a coordination mode was supported by the FT-IR spectra that exhibit well resolved band at 865cm(-1) and 1040cm(-1) attributable to ν(As-O) or ν(P-O) stretching vibration, respectively.

Influence of lead in the sorption of arsenate by municipal solid waste composts: metal(loid) retention, desorption and phytotoxicity

The ability of two municipal solid waste composts (MSW-C) to sorb As(V) in the presence of Pb(II) and in acidic conditions was investigated. Sorption isotherms and kinetics showed that both MSW-C were able to sorb As(V) in a similar way (∼0.24mmolg-1 MSW-C), but only when Pb(II) was present (0.45mmolL-1). The concomitant sorption of Pb(II) by both MSW-C (∼0.40mmolg-1) suggested that the metal cation was likely acting as bridging element between the negatively charged functional groups of composts and As(V). SEM-EDX analysis of the MSW-C+Pb(II)+As(V) systems supported the association between Pb(II) and As(V), while sequential extraction procedures and organic acids treatment showed that As(V) was strongly retained by MSW-C+Pb(II) and suggested the presence of different interaction types between As(V) and Pb(II). Plant growth experiments highlighted the key role of Pb(II) in the reduction of As(V)-phytotoxicity for triticale plants (×Triticosecale Wittm.) in the presence of MSW-C.

Role of polygalacturonic acid and the cooperative effect of caffeic and malic acids on the toxicity of Cu(II) towards triticale plants (× Triticosecale Wittm)

In this study, we investigated the cooperative effect of caffeic (CAF) and malic (MAL) acids as well as the role of polygalacturonic acid (PGA) on the toxicity of copper (Cu)(II) towards triticale plants (× Triticosecale Wittm.). Plant growth experiments carried out in hydroponic solutions at pH 6.0 revealed that CAF and PGA, but not MAL, were quite effective at reducing Cu(II) phytotoxicity and that MAL addition abolished the positive effects of CAF. The study of the CAF-Cu(II) system in aqueous solution clearly indicated that the lower toxicity of Cu(II) in the presence of CAF was due to the occurrence of a redox reaction between CAF and Cu(II), while the phytotoxicity recorded in the Cu(II)-CAF-MAL system was attributed to a competitive effect of MAL in the Cu(II) complexation by CAF, which ultimately reduced the redox activity. The lowest Cu(II) phytotoxicity recorded in the presence of PGA was attributed to the high affinity of the polysaccharidic matrix towards the metal ion. The results from this study shed new light on the Cu(II)-detoxifying action of selected root exudates and highlight the negative cooperation of MAL on the Cu(II)-detoxifying activity by CAF.

Arsenic mobilization by citrate and malate from a red mud-treated contaminated soil.

The mobility and bioavailability of As in the soil-plant system can be affected by a number of organic acids that originate from the activity of plants and microorganisms. In this study we evaluated the ability of citrate and malate anions to mobilize As in a polluted subacidic soil (UP soil) treated with red mud (RM soil). Both anions promoted the mobilization of As from UP and RM soils, with citrate being more effective than malate. The RM treatment induced a greater mobility of As. The amounts of As released in RM and UP soils treated with 3.0 mmol L citric acid solution were 2.78 and 1.83 μmol g respectively, whereas an amount equal to 1.73 and 1.06 μmol g was found after the treatment with a 3.0 mmol L malic acid solution. The release of As in both soils increased with increasing concentration of organic acids, and the co-release of Al and Fe in solution also increased. The sequential extraction showed that Fe/Al (oxi)hydroxides in RM were the main phases involved in As binding in RM soil. Two possible mechanisms could be responsible for As solubilization: (i) competition of the organic anions for As adsorption sites and (ii) partial dissolution of the adsorbents (e.g., dissolution of iron and aluminum oxi-hydroxides) induced by citrate or malate and formation of complexes between dissolved Fe and Al and organic anions. This is the first report on the effect of malate and citrate on the As mobility in a polluted soil treated with RM.

Assessment of the use potential of edible sea urchins (Paracentrotus lividus) processing waste within the agricultural system: Influence on soil chemical and biological properties and bean (Phaseolus vulgaris) and wheat (Triticum vulgare) growth in an amended acidic soil

In this study we evaluated the influence of ground purple sea urchin (Paracentrotus lividus) endoskeletons, a processing waste common to all edible sea urchin plants, on the chemical, biochemical and microbiological features of an acidic (pH 5.65) sandy-loam soil. The purple sea urchin endoskeletons were characterized by a high content of total carbonates (∼94%), a moderately alkaline pH in water (pH 7.88) and electrical conductivity values (3.55 mS/cm) very similar to those of commercial lime. To evaluate the influence of the P. lividus endoskeletons on soil properties four different amendment rates were tested, notably 0.5, 1.0, 3.0 and 5.0% based on soil dry weight, and the effects compared with those recorded on unamended control soil. The addition of the purple sea urchin processing waste caused an immediate and significant pH increase which was positively related to the rate of the amendment addition. After a six months equilibration period, the differences in soil pH were still evident and significant increases of electrical conductivity and available phosphorus were also detected in soils with the higher amendment rates. The number of heterotrophic and cellulolytic bacteria and actinomycetes significantly increased after amendment addition while the number of culturable fungi steadily declined. The analysis of the Biolog Community Level Physiological Profile indicated a clear influence of the purple sea urchin processing waste on the structure of the native microbial community while a significant increase of microbial functionality (i.e. dehydrogenase activity) was recorded in soil treated with the higher amendment rates (i.e. 3.0 and 5.0%). The improvement of microbial abundance and functionality as well as the change of the microbial community structure were ascribed to the pH shift induced by the P. lividus processing waste. To investigate possible effects on soil fertility, dwarf bean (Phaseolus vulgaris) and wheat (Triticum vulgare) growth were also assessed in a pot experiment. Plant growth was unaffected (wheat) or stimulated (bean) by the amendment addition in the 0.5-3.0% range while the higher amendment rate (i.e. 5.0%) was detrimental for both plant species indicating a phytotoxic effect which could be due to different factors such as an excess of calcium in soil, a suppression of Mg uptake or the higher EC values detected at the highest amendment rate. It is concluded that ground P. lividus endoskeletons have potential as a soil amendment to ameliorate chemical and biological properties of acidic Mediterranean soils. This seems particularly relevant, especially at the lower amendment rates, since for the first time, a sustainable management system is proposed for P. lividus processing waste, which foresees economic value in the sea urchin by-product through its re-use within the agricultural production system.