Mycoremediation of an agricultural salty soil contaminated with endosulfan by Penicillium crustosum: and agronomic bioassays with Phaseolus leptostachyus.

The fungus Penicillium crustosum was employed for endosulfan biodegradation, finding that sulphate endosulfan and mono alcohol endosulfan were the main compounds produced; therefore, an oxidative degradation pathway was suggested. A 93 ± 4.7% of Endosulfan degradation after one month of treatment of a highly salty agricultural soil was obtained, where ΔST was up to 17 ± 0.58 mN m-1, (related to the water value of 72 mNm-1), that was induced by the fungus during soil mycoremediation Additionally, an improvement in soil quality (reduction of clay proportion and salinity, as well as an increase of soluble phosphorus, carbon content and organic matter) was observed during the mycoremediation treatment. The phytotoxicity of the pesticide on Phaseolus leptostachyus was evaluated in the soil without the fungus addition (control), where the pesticide was translocated in the crop, presenting a negative effect in germination index, root length and weight, aerial weight, humidity, and proline content. This contrasted with the effect on the crop grown in the soil treated with P. crustosum, which had better agronomic characteristics. This is first report in which the effect of this property allows the pesticide biodegradation, due to a combined Endosulfan bioavailability and fungal biodegradation.

Measuring quantitative proteomic distance between Spanish beef breeds.

Estimates of quantitative proteomic distance between populations have not been reported to date. Here, quantitative proteomic distances between three Spanish bovine breeds (Asturiana de los Valles, AV; Retinta, RE; and Rubia Gallega, RG) were estimated from two-dimensional electrophoresis profiles of meat samples of longissimus thoracis muscle at 2 h post-mortem. Statistically significant distances were detected between AV/RG and the most genetically different RE breed, using the novel QD measure of quantitative proteomic distance. In total, 18 differentially abundant myofibrillar and sarcoplasmic proteins/isoforms contributing to proteomic distances between breeds were confidently identified by tandem mass spectrometry. The fast skeletal myosin regulatory light chain 2 followed by other five interacting proteins exhibited the most pronounced relative change between breeds. In addition, most differentially represented proteins could be associated with variations in meat tenderness. Therefore, they could be candidate biomarkers for molecular breeding programs and authentication of the three Spanish beef breeds.

Azo dyes decolorization under high alkalinity and salinity conditions by Halomonas sp. in batch and packed bed reactor.

Biodecolorization and biodegradation of azo dyes are a challenge due to their recalcitrance and the characteristics of textile effluents. This study presents the use of Halomonas sp. in the decolorization of azo dyes Reactive Black 5 (RB5), Remazol Brilliant Violet 5R (RV5), and Reactive Orange 16 (RO16) under high alkalinity and salinity conditions. Firstly, the effect of air supply, pH, salinity and dye concentration was evaluated. Halomonas sp. was able to remove above 84% of all dyes in a wide range of pH (6-11) and salt concentrations (2-10%). The decolorization efficiency of RB5, RV5, and RO16 was found to be ≥ 90% after 24, 13 and 3 h, respectively, at 50 mg L-1 of dyes. The process was monitored by HPLC-DAD, finding a reduction of dyes along the time. Further, Halomonas sp. was immobilized in volcanic rocks and used in a packed bed reactor for 72 days, achieving a removal rate of 3.48, 5.73, and 8.52 mg L-1 h-1, for RB5, RV5 and RO16, respectively, at 11.8 h. The study has confirmed the potential of Halomonas sp. to decolorize azo dyes under high salinity and alkalinity conditions and opened a scope for future research in the treatment of textile effluents.

Investigating the mechanisms underlying phytoprotection by plant growth-promoting rhizobacteria in Spartina densiflora under metal stress.

Pollution of coasts by toxic metals and metalloids is a worldwide problem for which phytoremediation using halophytes and associated microbiomes is becoming relevant. Metal(loid) excess is a constraint for plant establishment and development, and plant growth promoting rhizobacteria (PGPR) mitigate plant stress under these conditions. However, mechanisms underlying this effect remain elusive. The effect of toxic metal(loid)s on activity and gene expression of ROS-scavenging enzymes in roots of the halophyte Spartina densiflora grown on real polluted sediments in a greenhouse experiment was investigated. Sediments of the metal-polluted joint estuary of Tinto and Odiel rivers and control, unpollutred samples from the Piedras estuary were collected and submitted to ICP-OES. Seeds of S. densiflora were collected from the polluted Odiel marshes and grown in polluted and unpolluted sediments. Rhizophere biofilm-forming bacteria were selected based on metal tolerance and inoculated to S. densiflora and grown for 4 months. Fresh or frozen harvested plants were used for enzyme assays and gene expression studies, respectively. Metal excess induced SOD (five-fold increase), whereas CAT and ascorbate peroxidase displayed minor induction (twofold). A twofold increase of TBARs indicated membrane damage. Our results showed that metal-resistant PGPR (P. agglomerans RSO6 and RSO7 and B. aryabhattai RSO25) contributed to alleviate metal stress, as deduced from lower levels of all antioxidant enzymes to levels below those of non-exposed plants. The oxidative stress index (OSI) decreased between 50 and 75% upon inoculation. The results also evidenced the important role of PAL, involved in secondary metabolism and/or lignin synthesis, as a pathway for metal stress management in this halophyte upon inoculation with appropriate PGPR, since the different inoculation treatments enhanced PAL expression between 3.75- and five-fold. Our data confirm, at the molecular level, the role of PGPR in alleviating metal stress in S. densiflora and evidence the difficulty of working with halophytes for which little genetic information is available.

Complete Genome Sequence of Human Coronavirus OC43 Isolated from Mexico.

We report the complete genome sequence of the first Mexican human coronavirus (HCoV) OC43, obtained by new-generation sequencing and a metagenomic approach, isolated from a child hospitalized with pneumonia. The genome is closely related to the other OC43 genome sequences available, ranging from 99.8% to 98.2% nucleotide sequence identity.

Effects of organic amendments on the mobility of Pb and Zn from mine tailings added to semi-arid soils.

The effects of mine tailings and three organic amendments (compost, bokashi and vermicompost) on the mobility factor for Pb and Zn and on the potential C and N mineralization in semi-arid agricultural and rangeland soils were examined. During the experiment, soil samples were analyzed periodically for CO(2)-C evolution, inorganic N, dehydrogenase activity and percent Pb and Zn mobility during 169 d of incubation. The dehydrogenase activity and CO(2)-C evolved were strongly inhibited by mine tailings mixed with organic compost in both agricultural and rangeland soils (37 to 43 %), followed by N mineralization in treatments with mine tailings plus bokashi or compost (13 to 26.5 %) at 169 d incubation. The highest % mobility of Pb and Zn were observed in soils amended with mine tailings alone, while the lowest was observed in agricultural soils treated with vermicompost plus mine tailings. The CO(2)-C evolved was fitted to first order E, while the cumulative N mineralization was fitted to the linearized power function. Mine tailings were found to influence the potential C and N mineralization rate constants in both soils. The models for C and N mineralization could be used to evaluate the effects of mine tailings, which include intrinsic parameters in the soil.

PCBs stimulate laccase production and activity in Pleurotus ostreatus thus promoting their removal.

Pleurotus ostreatus degrades polychlorinated biphenyls (PCBs) with an increase of laccase activity. Laccases are well known for their detoxifying activity. We show, using reverse transcription polymerase chain reaction and a biochemical assay, that reduction in PCBs (di, tri, tetra, and penta) levels are correlated with an increase in laccase activity. P. ostreatus cultures were obtained from 0 to 30 days in the presence or absence of 7,100 mg/L PCBs (from transformer oil) and a surfactant. After each selected time cultures were withdrawn and remaining PCBs were determined, a maximal removal percentage of PCBs was obtained at 20 (63.5 ± 2.0) and 30 days (63.8 ± 4.6) post-induction. Also, the activity of the enzyme was analyzed and it was found to increase at 10 (6.9-fold) and 20 (6.77-fold) days post-induction in the presence of PCBs, as determined by its activity. Taken together, these data suggest that PCBs induce laccase expression and that laccase catalyzes PCBs removal.

Fractionation and availability of heavy metals in tannery sludge-amended soil and toxicity assessment on the fully-grown Phaseolus vulgaris cultivars.

This study was conducted to assess the effect of tannery sludge on the bush bean (Phaseolus vulgaris) cultivars fully-grown on a culture sandy soil, as tannery sludge is valuable to improve soil fertility but long term studies evaluating the effect on fully grown plants are scarce. Tannery sludge amendments (0, 0.77, 1.54, 3.08 and 6.16 g tannery sludge kg(-1) soil) were characterized and the main heavy metals identified (Cr, Mn, Fe, K, and Zn) later on sequentially and singly extracted, for soil fractionation and availability determination, respectively. Metals showed different fractionation and availability patterns, being the most toxic metal (Cr) found to primarily bind to the carbonate fraction in soil, while almost 10% of the total Cr was available for plant uptake. In the green house experiments, bush bean cultivars exposed to increasing tannery sludge amendments were evaluated at different plant stages. Metal accumulation and physiological parameters (chlorophyll, carotenoids, nitrate reductase activity and dry weight) were determined. Toxicity was primarily due to Cr, stimulating or affecting the response of physiological parameters and suppressing seed formation at the highest tannery sludge ratio. Metals were mainly accumulated in the roots of bush beans, diminishing in the upper part of the plants with minimal translocation to seeds, supposing little risk for human consumption. Additionally, important correlations, antagonistic and synergistic relationships were observed between the extracted metals and metal accumulation in plant tissues.

Toxicity assessment of soil amended with tannery sludge, trivalent chromium and hexavalent chromium, using wheat, oat and sorghum plants.

This work assessed the effect of soil amended with tannery sludge (0, 500, 1000, 2000, 4000 and 8000 mg Cr kg(-1)soil), Cr(3+) as CrCl(3).6H(2)O (0, 100, 250, 500, 1000 and 2000 mg Cr kg(-1)soil), and Cr(6+) as K(2)Cr(2)O(7) (0, 25, 50, 100, 200 and 500 mg Cr kg(-1)soil) on wheat, oat and sorghum plants. Seed germination, seedling growth (root and shoot) and Cr accumulation in dry tissue were measured. Toxicological parameters; medium effective concentration, no observed adverse effect concentration and low observed adverse effect concentration were determined. Root growth was the most sensitive assessment of Cr toxicity (P<0.05). There was a significant correlation (P<0.0001) between Cr accumulation in dry tissue and toxic effects on seedling growth. The three Cr sources had different accumulation and mobility patterns; tannery sludge was less toxic for all three plant species, followed by CrCl(3).6H(2)O and K(2)Cr(2)O(7).

Chromium fractionation in semi-arid soils amended with chromium and tannery sludge.

We studied Cr fractionation in three semi-arid soils (cultivated, under-the-canopy, and outside-the-canopy soils). The soils were amended with: Cr3+, Cr6+, tannery sludge, Cr3++tannery sludge, and Cr6++tannery sludge and all soils were incubated for 30 and 120 days at 25 degrees C. The Cr in three semi-arid soils was fractionated using sequential extraction (Tessier scheme). Data of Cr fractionation were used to find the correlations with microbial activities determined in previous work. The microbial activities: CO2-C evolved, dehydrogenase activity and nitrification were determined in the same soils amended with the same treatments. Tannery sludge was added at 0.0125 g g(-1) soil and Cr3+ or Cr6+ at 250 microg g(-1) soils. After 120 days of incubation, higher values of concentration of Cr were found in the residual fraction in the three soils amended with all the treatments, except cultivated and outside-the-canopy soils amended with Cr6++tannery. The non-residual fraction tended to decrease with time except in cultivated and outside-the-canopy soils amended with Cr6++tannery sludge. CO2-C evolved was significantly correlated (p<0.05 and p<0.001) with fractions bound to: Mn oxides, Fe oxides organic matter at 30 and 120 days of incubation (from r=0.827 to 0.979). Dehydrogenase activity was correlated with fractions bound to Fe oxides and bound to organic matter, and nitrification with fraction bound to organic matter at 30 days of incubation (r=0.874, 0.959, and 0.803, respectively). These results suggest that even in a sparingly available Cr fraction in semi-arid soils has effect on microbial activities.

Accumulation and localization of cadmium in Echinochloa polystachya grown within a hydroponic system.

Phytoremediation is a technology for extracting or inactivating pollutants. Echinochloa polystachya [(H.B.K.) Hitchcock] (Poaceae) is a fast-growing perennial grass that is common in tropical areas and is often found in oil-polluted soils that contain high concentrations of heavy metals. However, its tolerance to heavy metals, and its ability to accumulate them, has yet to be investigated. Here we test the hypothesis that E. polystachya is able to accumulate high concentrations of cadmium (Cd). Plants were grown hydroponically with different levels of Cd(2+) (0, 0.25, 1, 2, 10, 50, and 100mgL(-1)), and were found to be tolerant to Cd(2+) at all levels. No metal-toxicity symptoms were observed at any Cd(2+) level. Root and leaves Cd concentrations were 299+/-13.93 and 233+/-8.77mgkg(-1) (on a dry weight basis), respectively. Scanning electron microscopy showed the inclusion of Cd within the xylem; this result was confirmed by energy dispersive X-ray spectrometry. Leaf tissues also accumulated Cd, especially within the bulliform cells of the epidermis. We conclude that E. polystachya is a hyperaccumulator of Cd. While data for other metals are not yet available, E. polystachya shows promise in the phytoextraction of Cd from polluted tropical sites.

Effect of culture conditions on fatty acids composition of a biosurfactant produced by Candida ingens and changes of surface tension of culture media.

This work investigated the effect of culture medium composition on a biosurfactant production and their total fatty acids content, as well as the surface tension of media, and biomass production by Candida ingens. A factorial experimental design was used to evaluate the combined effect of C/P, C/N(inorganic), C/Fe, C/Mg ratios and yeast extract concentration. The highest biosurfactant production was reached when high C/Fe and high C/P ratio variables were combined; biosurfactant concentration increased by a 3.42 fold. The variable with the highest effect on net decrease surface tension (DeltaST) and fatty acids percentage of C. ingens biosurfactant was yeast extract. The average of DeltaST (25 mN/m) and fatty acids percentage (34.7%) values were enhanced at high yeast extract concentration of 1g/l. The main conclusion of this study was that the culture composition affected the biosurfactant production by C. ingens. It was also observed that the surface tension and total fatty acids of the biosurfactant were modified as the media composition changed.

Phenanthrene removal by Penicillium frequentans grown on a solid-state culture: effect of oxygen concentration.

Phenanthrene removal by Penicillium frequentans was compared under aerobic and microaerophilic conditions in a solid culture amended with low quantities of an agricultural residue. An inoculum of P. frequentans grown on sugarcane bagasse pith was mixed with soil spiked with 200 mg l(-1) of phenanthrene, to obtain a final bagasse/soil ratio of 1:16. The C/N ratio was adjusted to 60 and the moisture content to 40%. The oxygen concentrations were adjusted to 20%, 10%, 5%, 2% and close to 0%, in the soil-gas phase for each treatment. There were statistically significant (p<0.05) differences in the metabolic activity at different oxygen concentrations, measured as CO2 production. Phenanthrene removal rates increased with oxygen concentration, reaching 52% removal after 17 days of incubation for the treatment with 20% O2. Nevertheless, oxygen-limited (microaerophilic) conditions did not preclude phenanthrene degradation.

Effects of mixing low amounts of orange peel (Citrus reticulata) with hydrocarbon-contaminated soil in solid culture to promote remediation.

The effect of mixing low amounts of orange peel (Citrus reticulata) with a soil contaminated with hydrocarbons (58,000 mg kg(-1) soil) for promoting the soil remediation in solid culture was studied. The experimental design was established in solid culture at soil/orange (Citrus reticulata) peel ratios of 100:0, 98:2, 96:4, 94:6 and 92:8, at 30% humidity and a C:N:P ratio of 100:10:1, for 15, 60 and 90 days, respectively. The total petroleum hydrocarbons (TPHs) decreased significantly (69%) after 15 days in the treatment with a soil to orange peel ratio of 92:8. Furthermore, in this treatment bacterial counts increased from 17 to 20 ln CFU (2.6 x 10(6) to 5 x 10(8)), while the fungal count was 11 ln CFU (6.5 x 10(4)) at initial and final time of treatment. An increase in microbial respiration activity and TPH removal (69%) was observed at other soil/orange peel ratios after 60 days when moisture content and nutrients were adjusted; however, N and P were not consumed at a great extent.

Role of phenanthrene in rhamnolipid production by P. putida in different media.

The role of phenanthrene in rhamnolipid production by P. putida in eight media with different culture conditions was investigated. Cultures using Fe2SO4.7H2O, KH2PO4, NH4Cl, yeast extract, glucose, and corn oil, with and without 200 mg l(-1) of phenanthrene, were evaluated under shaking for rhamnolipid production through a 2(7-4) fractional factorial experimental design. The biosurfactant production, decrease in the surface tension of the broth and the total cell growth in media without phenanthrene were affected significantly (p < 0.001) by yeast extract, glucose, corn oil and NH4Cl, and in media with phenanthrene by glucose and yeast extract. The non polar fraction of the biosurfactant in all media was composed of linoleic (C18:2), arachidic (C20:0) and behenic (C22:0) fatty acids. The medium with phenanthrene (200 mg l(-1)), Fe2SO4.7H2O (5 x 10(-4) g l(-1)), KH2PO4 (0.2 g l(-1)), glucose (50 g l(-1)), yeast extract (1 g l(-1)), corn oil (2% vol), and NH4Cl (1 g l(-1)), shaken at 150 rpm at 37 degrees C, and pH 7.0, presented the highest biosurfactant production. For this medium the surface tension decreased by 35.9 mN m(-1) in relation to the initial value, and only this medium showed an emulsion capacity of 20%. The polar fraction (Rhamnose) in media 1, 3, 7 and 8 with phenanthrene was c.a 100%, in contrast to those without phenanthrene where this fraction was undetectable.

Selection and identification of fungi isolated from sugarcane bagasse and their application for phenanthrene removal from soil.

This work investigated the identification and selection of fungi isolated from sugarcane bagasse and their application for phenanthrene (Phe) removal from soil. Fungi were identified by PCR amplification of ITS regions as Aspergillus terrus, Aspergillus fumigatus and Aspergillus niger, Penicillium glabrum and Cladosporium cladosporioides. A primary selection of fungi was accomplished in plate, considering Phe tolerance of every strain in two different media: potato dextrose agar (PDA) and mineral medium (MM). The radial extension rate (r(r)) in PDA exhibited significant differences (p<0.05) at 200 and 400 ppm of Phe. A secondary selection of A. niger, C. cladosporoides, and P. glabrum sp. was achieved based on their tolerance to 200, 400, 600 and 800 ppm of Phe, in solid culture at a sugarcane bagasse/contaminated soil ratio of 95:5, in Toyamas, Czapeck and Wunder media. Under these conditions, a maximum (70%) Phe removal by A. niger was obtained. In addition C. cladosporioides and A. niger were able to remove high (800 ppm) Phe concentrations.

Bioavailable cadmium during the bioremediation of phenanthrene-contaminated soils using the diffusive gradients in thin-film technique.

AIMS: To study the impact of fungal bioremediation of phenanthrene on trace cadmium solid-solution fluxes and solution phase concentration. METHODS AND RESULTS: The bioremediation of phenanthrene in soils was performed using the fungus Penicillium frequentans. Metal behaviour was evaluated by the techniques of diffusive gradient in thin-films (DGT) and filtration. Fluxes of cadmium (Cd) show a significant (P < 0.002) increase after the start of bioremediation, indicating that the bioremediation process itself releases significant amount of Cd into solution from the soil solid-phase. Unlike DGT devices, the solution concentration from filtration shows a clear bimodal distribution. We postulate that the initial action of the fungi is most likely to breakdown the surface of the solid phase to smaller, 'solution-phase' material (<0.45 microm) leading to a peak in Cd concentration in solution. CONCLUSIONS: Phenanthrene removal from soils by bioremediation ironically results in the mobilization of another toxic pollutant (Cd). SIGNIFICANCE AND IMPACT OF THE STUDY: Bioremediation of organic pollutants in contaminated soil will likely lead to large increases in the mobilization of toxic metals, increasing metal bio-uptake and incorporation into the wider food chain. Bioremediation strategies need to account for this behaviour and further research is required both to understand the generality of this behaviour and the operative mechanisms.

Benzo[a]pyrene removal from soil by Phanerochaete chrysosporium grown on sugarcane bagasse and pine sawdust.

The capacity of Phanerochaete chrysosporium grown on soil with added sugarcane baggase (BP) and pine sawdust (PS) to remove benzo(a)pyrene (BaP) was studied. A half factorial two-level experiment 2(4-1) was designed to determine the effect of: type of lignocellulosic material (BP and PS) for fungus growth, age of fungus (5 and 10d), amount of lignocellulosic material (10% and 15% w/w) and soil moisture content (water holding capacity of 45% and 56% w/w). Inoculum obtained at different ages showed that the capacity of P. chrysosporium to remove BaP depends on the lignocellulosic used and on inoculum age. Abiotic BaP removal was affected significantly (p<0.05) by inoculum age, type of lignocellulosic added and soil moisture content. The removal of BaP by lignocellulosic material was more effective by young inocula (71.97 mg BaP kg(-1) dry soil), with high percentage of added lignocellulosic (71.57 mg BaP kg(-1) dry soil) and at low soil moisture content (73.07 mg BaP kg(-1) dry soil). When fungus was grown on BP, maximum BaP removal rate was obtained at 5d of incubation (10.85 mg BaP d(-1)l(-1) and 50.12 mg BaP kg(-1) dry soil), while in PS maximum BaP removal was obtained at 10d of incubation (12.06 mg BaP d(-1)l(-1) and 39.94 mg BaP kg(-1) dry soil).

Effect of combined nutrients on biosurfactant produced by Pseudomonas putida.

This work investigated biosurfactant production by Pseudomonas putida in combined C/P, C/Ninorganic, C/Fe, C/Mg nutrient ratios and peptone concentration. Analysis of the 2(5-1) fractional factorial experimental design showed that only the C/Fe ratio had a significant (p<0.02) effect on biosurfactant production. The highest amount of biosurfactant was obtained at low C/Fe ratios, but net surface tension did not show significant differences. In addition, low amounts of peptone and the C/P-C/Mg nutrient ratios interaction significantly (p < 0.05) enhanced the biomass produced by P. putida. Analysis of biosurfactant by gas chromatography (GC) showed that the hydrophilic fraction was composed by rhamnose and the hydrophobic fraction, mainly by palmitic (C16), stearic (C18:0), oleic (C18:1) and linoleic (C18:2) fatty acids.