Pressurized liquid extraction and HPLC quantification of folic Acid in fortified wheat flours

A pressurized liquid extraction (PLE) method using phosphate buffer as solvent was applied for folic acid (FA) extraction from fortified wheat flours and was compared to a standard solid-liquid extraction (SLE) method. Extracted FA was quantified by reverse phase high-performance liquid chromatography (RP-HPLC) hyphenated with a phenyl column and an absorption photometric detector (λ = 280 nm). Detection and quantification limits were 0.12 and 0.4 ng, respectively, corresponding to 0.06 and 0.2 μg g(-1) of analyzed wheat flour. Equivalent FA contents were found by both extraction methods, but a single PLE allowed a total recovery of FA content, whereas at least three successive SLEs were needed to achieve a total recovery of FA. The obtained results indicated that PLE is a rapid and efficient technique for FA extraction from fortified wheat flour.     

Influence of Desiccation on the Transport of <Emphasis Type="Italic">Escherichia coli</Emphasis> through Saturated Sand Packs

Bacterial cells that enter the groundwater system commonly experience desiccation stresses (i.e., bacterial cells are directly exposed to air) when traveling through the unsaturated layer of soil. Little is known about the effects of desiccation on the transport of bacterial cells in the groundwater system. In this research, we investigated the transport of desiccated and non-desiccated Escherichia coli K12 (ATCC 10798) cells through saturated sand packs using laboratory column transport experiments. Cell desiccation was performed at 25°C under relative humidity (RH) levels of 22%, 53%, 75%, and 97%, respectively, and the desiccation duration was 22 h. Our results showed that desiccation reduced the viability of E. coli cells under all RH levels and increased the transport of E. coli cells under ≥75% RH levels. The increase in the transport of the desiccated E. coli cells was not related to changes in cell size or cell zeta potential. Desiccation under high (i.e., ≥75%) RH levels, however, led to lower cell hydrophobicity, which was found to be positively correlated with cell transport.     

The Production of Ligninolytic Enzymes by Marine-Derived Basidiomycetes and Their Biotechnological Potential in the Biodegradation of Recalcitrant Pollutants and the Treatment of Textile Effluents

Filamentous fungi derived from marine environments are well known as a potential genetic resource for various biotechnological applications. Although terrestrial fungi have been reported to be highly efficient in the remediation of xenobiotic pollutants, fungi isolated from the marine environment may possess biological advantages over terrestrial fungi because of their adaptations to high salinity and pH extremes. The present study describes the production of ligninolytic enzymes under saline and non-saline conditions and the decolorization of Remazol Brilliant Blue R (RBBR) dye by three basidiomycetes recovered from marine sponges (Tinctoporellus sp. CBMAI 1061, Marasmiellus sp. CBMAI 1062, and Peniophora sp. CBMAI 1063). Ligninolytic enzymes were primarily produced by these fungi in a salt-free malt extract and malt extract formulated with artificial seawater (saline condition). CuSO₄ and wheat bran were the best inducers of lignin peroxidase and manganese peroxidase activity. RBBR was decolorized up to 100% by the three fungi, and Tinctoporellus sp. CBMAI 1061 was the most efficient. Our results revealed the biotechnological potential of marine-derived basidiomycetes for dye decolorization and the treatment of colored effluent as well as for the degradation of other organopollutants by ligninolytic enzymes in non-saline and saline conditions that resemble the marine environment.     

Metal Tolerance, Accumulation and Translocation in Poplar and Willow Clones Treated with Cadmium in Hydroponics

To evaluate the phytoremediation capability of some poplar and willow clones a hydroponic screening for cadmium tolerance, accumulation and translocation was performed. Rooted cuttings were exposed for 3 weeks to 50 μM cadmium sulphate in a growth chamber and morpho-physiological parameters and cadmium content distribution in various parts of the plant were evaluated. Total leaf area and root characteristics in clones and species were affected by cadmium treatment in different ways. Poplar clones showed a remarkable variability whereas willow clones were observed to be more homogeneous in cadmium accumulation and distribution. This behaviour was further confirmed by the calculation of the bio-concentration factor (BCF) and the translocation factor (Tf). Mean values of all the clones of the two Salicaceae species showed that willows had a far greater ability to tolerate cadmium than poplars, as indicated by the tolerance index (Ti), calculated on the dry weight of roots and shoots of plants. As far as the mean values of Tf was concerned, the capacity of willows to translocate was double that of poplars. On the contrary, the mean values of total BCF in poplar clones was far higher with respect to those in willows. The implications of these results in the selection of Salicaceae clones for phytoremediation purposes were discussed.     

Factors affecting the contents of iridoid oleuropein in olive leaves (Olea europaea L.)

In this study, for the first time, the impact of the genetic factor on the contents of oleuropein in olive leaves was not only evaluated but the influence exerted by the color/age of leaves (green, green-yellowish, and yellow) and the collecting period (spring or autumn) was also evaluated. A repetitive high-resolution gas chromatographic quantitation method and an accurate high-performance liquid chromatographic method were developed. These analytical methods gave results showing a highly linear relationship. Samples of olive leaves were taken from seven major Italian olive cultivars, such as Dritta, Leccino, Caroleo, Coratina, Castiglionese, Nebbio, and Grossa di Cassano. Such a vegetal raw material could actually be exploited for recovering oleuropein, considered to be a high-added value molecule. This could be converted into hydrxytyrosol, a compound known to possess strong bioactive properties. Olive leaves showed considerable contents of oleuropein, which with some cultivars were even higher with respect to those present in the corresponding olive fruits (reported in the literature). The amounts of oleuropein in the collected leaves were markedly modified by the color/age and genetic factors, whereas meaningless variations were ascribable to the quantitation method and the collecting period factors. Various chemometrics, applied to the obtained analytical data, appeared to be effective in discriminating the samples on the basis of the above-examined experimental factors, thus confirming how these should be taken into account in future industrial recovery of oleuropein from olive leaves.     

Enhancement of reproductive performance in shrimp Litopenaeus vannamei (Boone, 1931) by supplementation of Ulva clathrata meal in maturation diet in two commercial hatcheries

The effect of supplementing dehydrated seaweed Ulva clathrata (20 g/kg) in the squid fraction of a formulated fresh diet on the reproductive performance of Litopenaeus vannamei broodstock was evaluated in two commercial hatcheries (A and B). The origin of broodstock, use of ablation on females, tank/culture conditions (for maturation, spawning, and larval rearing) were different between hatcheries. Daily mortality was higher (p < .01) for females in the control group compared to Ulva‐fed group of hatchery A, where females were ablated. No differences in mortality were found for unablated females in hatchery B, or with males from either hatchery. In hatchery A, broodstock fed Ulva produced more eggs (an increase in +5% compared to control, p < .05) and nauplii per female (+18%, p < .01). In hatchery B, broodstock fed Ulva produced significantly more eggs (+26%, p < .05) and nauplii per day (+33%, p < .05). In hatcheries A and B, eggs from broodstock fed Ulva had increased hatching rates (+13 and +7%, respectively, p < .001 in both cases). The number of spawns per day had a tendency to be higher in broodstock fed Ulva in both hatcheries, but the difference did not reach statistical significance (+14% for hatchery A and +20% for hatchery B). Dehydrated U. clathrata appears therefore as a functional additive in L. vannamei maturation diets that enhances the reproductive performance at commercial scale, regardless of the management conditions.     

Eco-physiological and Antioxidant Responses of Holm Oak (<Emphasis Type="Italic">Quercus ilex</Emphasis> L.) Leaves to Cd and Pb

Plants of the urban environment are exposed to a wide range of pollutants, including heavy metals. This research studies in situ the eco-physiological and antioxidant responses of holm oak (Q. ilex) leaves to Pb and Cd to assess the mechanisms of metal tolerance in this species, widely used as biomonitor. Leaves of plants grown at parks and roadsides were analyzed for photosynthetic activity, Pb and Cd concentration in tissues and cell-free extracts, thiol groups, D1 and Rubisco protein content, ascorbic acid (AsA) amount, and catalase (CAT) activity. The main results evidenced that Cd concentration was higher in leaves collected at the park out from the downtown; whereas Pb was most abundant in leaves sampled at the roadside nearby the highway. Pb in cell-free extracts was higher in park than in roadside leaves. Although Cd in the leaf tissues was twofold lower than Pb, it was more abundant than Pb in cellular extracts deprived of all particulate matter. Leaves responded to different concentration of Cd and Pb modulating some eco-physiological and biochemical traits, roadside leaves showed reduced leaf lamina, higher content of photosynthetic pigments, hydrogen peroxide, and AsA, as well as higher CAT activity compared to park leaves. In the roadside leaves, a stress condition for photosynthetic apparatus can be hypothesized on the basis of the decline of photochemical activity, the increase of NPQ, and the reduction of Rubisco and D1 protein content. The elevated presence of thiol groups in these leaves suggests a possible role of Pb and Cd in activation of antioxidant responses.     

Physiological response of culture media-grown barley (Hordeum vulgare L.) to titanium oxide nanoparticles

Since the fate of nanoparticles after their release in the environment and their possible transfer in plants and subsequent impacts is still largely unknown, this paper evaluates the potential phytotoxic effects of up to 20% w/w TiO₂ nanoparticles (nTiO₂) on barley cultivated in hydroponics and agar media. The X-ray diffraction analysis confirmed that nTiO₂ powder corresponds to anatase phase. On agar medium only high concentrations of nTiO₂ (10% and 20% w/w) induced significant inhibition of shoot growth. However, hydroponics treatment with nTiO₂ up to 1000?mg?L^?1 did not show any adverse effect on the shoot growth. In both experiments, (i) root growth inhibition effects became visible with increasing concentration of nTiO₂, (ii) plants treated with nTiO₂ showed no change in chlorophyll a and b content, even though the plants absorbed nTiO₂, (iii) weight of biomass treated with nTiO₂ was not significantly different compared to control. Therefore, we assume that transport of nTiO₂ into the aerial parts is limited due to the presence of effective mechanical or physiological barriers in roots. Overall, it appears that early root growth is a relevant indicator of potential effects of nTiO₂ exposure. Our results also indicate that synthesized nTiO₂ are not significantly toxic to the barley when applied at the concentrations used in this work, even though plants absorb titanium.