Biochemical responses of Echinochloa polystachya inoculated with a Trichoderma consortium during the removal of a pyrethroid-based pesticide.

The biochemical response of plants exposed to pesticides and inoculated with microorganisms is of great importance to explore cleaning up strategies for contaminated sites with pyrethroid-based pesticides. We evaluated the effects of a Trichoderma consortium on the biochemical responses of Echinochloa polystachya plants during the removal of a pyrethroid-based pesticide. Plants were inoculated or not with the Trichoderma consortium and exposed to commercial pesticide H24®, based on pyrethroids. Pesticide application resulted in significant reduction in root protein content (58%), but enhanced content of malondialdehyde (MDA) in shoots, superoxide dismutase (SOD) activity in shoots and roots, and catalase (CAT) activity in roots. Inoculation of Trichoderma consortium in E. polystachya exposed to the pesticide resulted in increased protein content in roots and MDA content in shoots (2-fold). Trichoderma consortium improved protein content and SOD activity (140-fold) in plants. Fungal inoculation increased the removal (97.9%) of the pesticide in comparison to the sole effect of plants (33.9%). Results allow further understanding about the responses of the interaction between plants and root-associated fungi to improving the assisted-phytoremediation of solid matrices contaminated with organic pesticides.

This original paper describes the positive role of the Trichoderma sp. consortium on favoring the removal of a pyretrhoid-based pesticide. This is one of first reports on analyzing the influence of a Trichoderma consortium on the oxidative stress and antioxidant response of Echinochloa polystachya in presence of the pesticide. This experimental approach provides a new alternative for further fungal assisted-phytoremediation of a pyretrhoid-based pesticide.

Phytoremediation of soil contaminated with weathered petroleum hydrocarbons by applying mineral fertilization, an anionic surfactant, or hydrocarbonoclastic bacteria.

This study evaluated the effect of the application of mineral fertilization (F), the anionic surfactant Triton X-100 (TX100), or the inoculation with a hydrocarbooclastic bacterial consortium (BCons) on the growth of Clitoria ternatea during the phytoremediation of a Gleysol contaminated with weathered petroleum hydrocarbons (39,000 mg kg-1 WPH) collected from La Venta, Tabasco (Mexico). The experiment consisted of a completely randomized design with seven treatments and four replications each under greenhouse conditions. The application of F (biostimulation) increased plant growth and biomass production; in contrast, TX100 only favored root biomass (11%) but significantly favored WPH degradation. Bioaugmentation with BCons did not show significant effects on plant growth. Nevertheless, the combination of biostimulation with bioaugmentation (BCons + F, BCons + TX100, and BCons + F+TX100) enhanced plant growth, hydrocarbonoclastic bacteria population, and WPH degradation when compared to treatments with the single application of bioaugmentation (BCons) or biostimulation (F).

Application of mineral fertilization and commercial surfactant favored root biomass and degradation of weathered petroleum hydrocarbons (WPH). The reintroduction of hydrocarbonoclastic and surfactant-producer bacteria did not enhance plant growth but significantly contributed on WPH degradation from a chronically contaminated soil.

Effect of Predation by Colpoda sp. in Nitrogen Fixation Rate of Two Free-Living Bacteria.

Biological nitrogen fixation is limited to several groups of prokaryotes, some of them reduce nitrogen as free-living nitrogen-fixing bacteria. Protozoa predation on these latter releases sequestered nitrogen that may enhance the formation of new bacterial biomass and possibly increase nitrogen fixation within soil microbial communities. We aim to evaluate the predation effect of Colpoda sp. on two nitrogen fixers: Azospirillum lipoferum and Stenotrophomonas sp. during their lag, early exponential, and exponential phases. The kinetics of bacterial population growth was determined in the predators' presence or absence and the effect of predation on the rate of N fixation was evaluated through the reduction of acetylene to ethylene technique. Colpoda sp. showed a non-significant difference in preferences between the two species offered as prey. Consequently, the abundance of A. lipoferum and Stenotrophomonas sp. decreased significantly due to predator's pressure and both species responded by increasing their specific growth rate. Likewise, predation promoted greater nitrogen fixation rate by CFU during the lag phase in A. lipoferum (0.20 nM/CFU with predation vs 0.09 nM/CFU without predation) and Stenotrophomonas sp. (0.22 nM/CFU vs 0.09 nM/CFU respectively). During early exponential phase (29 h), the rate diminished to 0.13 and 0.05 nM/CFU in A. lipoferum and to 0.09 nM/CFU and 0.05 nM/CFU in Stenotrophomonas sp. Finally, during the exponential phase (52 h), only A. lipoferum without predation produced 0.003 nM/CFU of ethylene. Thus, the nitrogenase activity was higher in the lag and the early exponential phases when predator activity was involved.

Efecto de bacterias emulsificantes en la atenuación de la fitotoxicidad de suelos contaminados con petróleo intemperizado / Effect of emulsifying bacteria on phytotoxicity attenuation of soils contaminated with weathered petroleum hydrocarbons

Introducción: Las plantas y los microorganismos se han utilizado como bioindicadores de la toxicidad inducida por hidrocarburos presentes en los suelos. Objetivo: El presente trabajo evaluó la toxicidad de un Gleysol contaminado de origen con diferentes concentraciones de petróleo intemperizado, recolectado en la Venta Tabasco (México), en el crecimiento de Clitoria ternatea, y la atenuación de la fitotoxicidad con la inoculación de bacterias emulsificantes. Metodología: Se usaron suelos con 50 y 150 g HTPI kg-1, y un suelo testigo con 0.15 g HTPI kg-1 (origen biogénico), y la inoculación de seis bacterias emulsificantes y su combinación (consorcio). La fitotoxicidad de los HTPI se evaluó considerando la altura, la biomasa seca (radical, aérea y total), el área foliar, el área foliar específica, y la eficiencia del fotosistema II (EPSII), a los 30 días. Resultados: Los HTPI no afectaron la altura, pero el suelo con 50 g HTPI kg-1 redujo la biomasa seca radical y total, y el área foliar con respecto a las plantas en los suelos testigo y con 150 g HTPI kg-1. La cepa Sml (Stenotrophomonas maltophilia C10S1) incrementó significativamente la biomasa seca total; la cepa Ro (Raoultella ornithinolyticaC5S3) produjo mayor área foliar específica con respecto a plantas no inoculadas. En el suelo testigo, el consorcio bacteriano estimuló la altura; las cepas Sm (Serratia marcescens C11S1) y Sm2 (S. marcescens C7S3) mejoraron la altura y el área foliar específica con respecto a plantas no inoculadas, en el suelo con 50 g HTPI kg-1. En el suelo con 150 g HTPI kg-1, las cepas Spa (Stenotrophomonas pavanii C5S3F) y Cfr (Citrobacter freundii C4S3) incrementaron la biomasa seca radical y aérea, respectivamente. La EPSII no fue afectada por la contaminación de los suelos. Las bacterias emulsificantes redujeron la fitotoxicidad de HTPI, pero dependiendo de su contenido en los suelos. Conclusiones: El suelo con 50 g HTPI kg-1 mostró mayor toxicidad en el crecimiento de las plantas. La inoculación bacteriana favoreció el crecimiento, producción de biomasa, y área foliar en el suelo con 150 g HTPI kg-1. La EPSII no fue afectada por la presencia de HTPI en el suelo.

Introduction: Plants and microorganisms have been used as bioindicators to evaluate the toxicity of hydrocarbons in soils. Objective: This study evaluates the toxicity of a chronically-contaminated Gleysol with several concentrations of weathered petroleum hydrocarbons (WPH), collected from La Venta, Tabasco (Mexico), on the growth of Clitoria ternatea and the phytoxicity attenuation due to inoculation of emulsifying bacteria. Methods: Soils with 50 and 150 g WPH kg-1, and control soil with 0.15 g WPH kg-1 (biogenic origin) were utilized, as well as the inoculation of six emulsifying bacteria and their combination (consortium). The WPH-phytotoxicity was evaluated by considering plant height, dry biomass production (root, shoot, and total), leaf area, specific leaf area, and the efficiency of photosystem II (EPSII), after 30 days. Results: WPH did not affect plant height, but soil with 50 g WPH kg-1 diminished root and total dry weight, and leaf area, when compared to both control soil and soil with 150 g WPH kg-1. The strain Sml (Stenotrophomonas maltophilia C10S1) significantly increased shoot and total dry weight, while the strain Ro (Raoultella ornithinolytica C5S3) produced higher specific leaf area relative to uninoculated plants. In control soil, the bacterial consortium stimulated plant height. The strains Sm (Serratia marcescens C11S1)and Sm2 (S. marcescens C7S3) improved plant height and specific leaf area when compared to uninoculated plants in soil with 50 g WPH kg-1. In soil with 150 g WPH kg-1, strains Spa (Stenotrophomonas pavanii C5S3F)and Cfr (Citrobacter freundii C4S3)enhanced root and shoot dry weight, respectively. The EPSII was unaffected by soil contamination. Emulsifying bacteria reduced the phytotoxic effects of WP, but depending on the content of WPH in soils. Conclusions: Soil with 50 g WPH kg-1 showed the greatest phytotoxic effects on plant growth. Bacterial inoculation favored growth, biomass production and leaf area in soil with 150 g WPH kg-1. The EPSII was not affected by WPH in soils.

Microbial Bioleaching of Ag, Au and Cu from Printed Circuit Boards of Mobile Phones.

Electronic waste (E-Waste) is consumed at high speed in the world. These residues contain metals that increase their price each year, generating new research on the ability of microorganisms to recover the metals from these wastes. Therefore, this work evaluated the biologic lixiviation of Cu, Ag and Au from printed circuit boards (PCB) of mobile phones by three strains of Aspergillus niger, Candida orthopsilosis, Sphingomonas sp. and their respective consortia, in addition to leaching with citric acid. The microorganisms were cultured in mineral media with 0.5 g of PCB, and the treatments with 1M citric acid were added the same amount of PCB. All treatments were incubated for 35 days at room temperature. The results showed that Sphingomonas sp. MXB8 and the consortium of C. orthopsilosis MXL20 and A. niger MXPE6 can increase their dry biomass by 147% and 126%, respectively, in the presence of PCB. In the bioleaching of metals, the inoculation of A. niger MXPE6, the consortium of Sphingomonas sp. MXB8/C. orthopsilosis MXL20 and Sphingomonas sp. MXB8 leached 54%, 44.2% and 35.8% of Ag. The consortium of A. niger MX5 and A. niger MXPE6 showed a leaching of 0.53% of Au. A. niger MX5 leaching 2.8% Cu. Citric acid increased Cu leaching by 280% compared to treatments inoculated with microorganisms. Although further research is required, A. niger MXPE6 and the consortium of Sphingomonas sp. MXB8/C. orthopsilosis MXL20 could be an alternative to recover Ag from PCB of mobile phones.

Lipid extraction from the biomass of Trichoderma koningiopsis MX1 produced in a non-stirring culture for potential biodiesel production.

Oleaginous microorganisms such as microalgae, yeasts, bacteria and filamentous fungi are alternative sources of vegetal or animal fats for biodiesel production. This research evaluated the lipid production by the biomass Trichoderma koningiopsis MX1 with a non-stirring culture at room temperature, and fungal lipids were extracted through three techniques for biodiesel generation purposes. The three modified lipid extraction techniques yielded 18.4, 10.3 and 17.1 % of fungal lipids. The trans-esterification of lipids indicated that the controlling components for biodiesel were palmitic (40.8 %) and linoleic acids (ranging from 37.6 % to 41.2 %). Results show that fungal cultural conditions and the lipid extraction technique are determinants for producing biodiesel from fungal lipids. Therefore, the modification of some of these conditions could increase their efficiency and viability.

Bioleaching of gold, copper and nickel from waste cellular phone PCBs and computer goldfinger motherboards by two Aspergillus nigerstrains.

In an effort to develop alternate techniques to recover metals from waste electrical and electronic equipment (WEEE), this research evaluated the bioleaching efficiency of gold (Au), copper (Cu) and nickel (Ni) by two strains of Aspergillus niger in the presence of gold-plated finger integrated circuits found in computer motherboards (GFICMs) and cellular phone printed circuit boards (PCBs). These three metals were analyzed for their commercial value and their diverse applications in the industry. Au-bioleaching ranged from 42 to 1% for Aspergillus niger strain MXPE6; with the combination of Aspergillus niger MXPE6 + Aspergillus niger MX7, the Au-bioleaching was 87 and 28% for PCBs and GFICMs, respectively. In contrast, the bioleaching of Cu by Aspergillus niger MXPE6 was 24 and 5%; using the combination of both strains, the values were 0.2 and 29% for PCBs and GFICMs, respectively. Fungal Ni-leaching was only found for PCBs, but with no significant differences among treatments. Improvement of the metal recovery efficiency by means of fungal metabolism is also discussed.

Bioleaching of gold, copper and nickel from waste cellular phone PCBs and computer goldfinger motherboards by two Aspergillus nigerstrains

In an effort to develop alternate techniques to recover metals from waste electrical and electronic equipment (WEEE), this research evaluated the bioleaching efficiency of gold (Au), copper (Cu) and nickel (Ni) by two strains of Aspergillus niger in the presence of gold-plated finger integrated circuits found in computer motherboards (GFICMs) and cellular phone printed circuit boards (PCBs). These three metals were analyzed for their commercial value and their diverse applications in the industry. Au-bioleaching ranged from 42 to 1% for Aspergillus niger strain MXPE6; with the combination of Aspergillus niger MXPE6 + Aspergillus niger MX7, the Au-bioleaching was 87 and 28% for PCBs and GFICMs, respectively. In contrast, the bioleaching of Cu by Aspergillus niger MXPE6 was 24 and 5%; using the combination of both strains, the values were 0.2 and 29% for PCBs and GFICMs, respectively. Fungal Ni-leaching was only found for PCBs, but with no significant differences among treatments. Improvement of the metal recovery efficiency by means of fungal metabolism is also discussed.

Trophic structure of amoeba communities near roots of Medicago sativa after contamination with fuel oil no. 6.

Root exudation increases microbial activity, selecting bacterial and fungal communities that metabolize organic matter such as hydrocarbons. However, a strong contamination pulse of hydrocarbons around plant roots may reorganize the soil's microbial trophic structure toward amoebae feeding on bacteria. We conducted a microcosm experiment to elucidate the effect of Medicago sativa on the trophic structure of naked amoebae after a strong pulse of pollution (50,000 ppm of fuel oil no. 6, which is a mixture of long chains ranging from C10 to C28). Plants were seeded 24 h after contamination and species of amoebae in the microcosms were identified at 1, 30, and 60 days after pollution. Several species from three trophic groups of naked amoeba were still alive 24 h after the hydrocarbon pulse. Non-planted microcosms harbored three trophic groups after 60 days, while planted ones nourished four groups. The bacterivore group was the most diverse in all microcosms, followed by protist-eaters and omnivores. The quantity of amoebae was significantly higher (3.4×10(3) organisms/g soil) in the planted pots than in the non-planted ones (1.3×10(3) organisms/g soil after 30 days of pollution (P ≤ 0.01). The shortest hydrocarbon chains (C10-C14) disappeared or diminished in all microcosms, and the longest ones increased in the planted ones. M. sativa thus exerted a positive effect on species richness, quantity, and the composition of amoebae trophic groups in contaminated soil. This indirect effect on bacterial predators is another key factor underlying hydrocarbon assimilation by living organisms during phytoremediation.

Arbuscular mycorrhizal fungi on growth, nutrient status, and total antioxidant activity of Melilotus albus during phytoremediation of a diesel-contaminated substrate.

This research evaluated the effects of arbuscular mycorrhizal fungi (AMF) on growth, nutritional status, total antioxidant activity (AOX), total soluble phenolics content (TPC), and total nitrate reductase activity (NRA) of leaves and roots of Melilotus albus Medik planted in diesel-contaminated sand (7500 mg kg(-1)). Seedlings of Melilotus either Non inoculated (Non-AMF) or pre-inoculated plants (AMF) with the AMF-inoculum Glomus Zac-19 were transplanted to non-contaminated or contaminated sand. After 60 days, diesel significantly reduced plant growth. AMF- plants had no significant greater (64% and 89%, respectively) shoot and leaf dry weight than Non-AMF plants, but AMF plants had lower specific leaf area. AMF-plants had significantly greater content of microelements than non-AMF plants. Regardless diesel contamination, the total AOX and TPC were significantly higher in leaves when compared to roots; in contrast, NRA was higher in roots than leaves. Diesel increased total AOX of leaves, but AMF-plants had significantly lower AOX than non-AMF plants. In contrast, roots of AMF-plants had significantly higher AOX but lower NRA than non-AMF plants. AMF-colonization in roots detected via the fungal alkaline phosphatase activity was significantly reduced by the presence of diesel. AMF-inoculation alleviated diesel toxicity on M. albus by enhancing plant biomass, nutrient content, and AOX activity. In addition, AMF-plants significantly contributed in higher degradation of total petroleum hydrocarbons when compared to non-AMF-plants.