Microbial diversity and bioremediation of rhizospheric soils from Trindade Island - Brazil.

Pristine environments may harbor complex microbial communities with metabolic potential for use in bioremediation of organic pollutants. This study aimed to evaluate crude oil biodegradation by microbial communities present in rhizospheric soils of Bulbostylis nesiotis and Cyperus atlanticus on Trindade Island and the compositional structure of these communities. After 60 days under aerobic conditions, Total Petroleum Hydrocarbon biodegradation ranged from 66 to 75%, depending on the plant species and the origin of the soil samples. There was no response of petroleum biodegradation to fertilization with N:P:K (80:160:80 mg dm-3). Soil contamination with crude oil did not necessarily reduce microbial diversity. The richness and diversity increased in contaminated soils in some specific situations. We conclude that microbial communities from pristine soils have the ability to remove hydrocarbons through biodegradation and that Bulbostylis nesiotis and Cyperus atlanticus inhabiting Trindade Island harbor rhizospheric microbial communities with potential for application in rhizoremediation.

Aliphatic Hydrocarbon Enhances Phenanthrene Degradation by Autochthonous Prokaryotic Communities from a Pristine Seawater.

The microbial diversity and functioning around oceanic islands is poorly described, despite its importance for ecosystem homeostasis. Here, we aimed to verify the occurrence of microbe-driven phenanthrene co-oxidation in the seawater surrounding the Trindade Island (Brazil). We also used Next-Generation Sequencing to evaluate the effects of aliphatic and polycyclic aromatic hydrocarbons (PAHs) on these microbial community assemblies. Microcosms containing seawater from the island enriched with either labelled (9-14C) or non-labelled phenanthrene together with hexadecane, weathered oil, fluoranthene or pyrene, and combinations of these compounds were incubated. Biodegradation of phenanthrene-9-14C was negatively affected in the presence of weathered oil and PAHs but increased in the presence of hexadecane. PAH contamination caused shifts in the seawater microbial community-from a highly diverse one dominated by Alphaproteobacteria to less diverse communities dominated by Gammaproteobacteria. Furthermore, the combination of PAHs exerted a compounded negative influence on the microbial community, reducing its diversity and thus functional capacity of the ecosystem. These results advance our understanding of bacterial community dynamics in response to contrasting qualities of hydrocarbon contamination. This understanding is fundamental in the application and monitoring of bioremediation strategies if accidents involving oil spillages occur near Trindade Island and similar ecosystems.

A new biostimulation approach based on the concept of remaining P for soil bioremediation.

C:N:P ratio is generally adopted to estimate the amount of nitrogen and phosphorus to be added to soils to accelerate biodegradation of organic contaminants. However, differences in P fixation among soils lead to varying amounts of available P when a specific dose of the element is applied to different soils. Thus, the application of fertilizers to achieve a previously established C:P ratio leads to biodegradation rates that can be lower than the theoretical maximum. In this study, we developed an equation to estimate the dose of P required to maximize organic contaminant biodegradation in soils as a function of remaining P (P-rem), using diesel as a model contaminant. The soils were contaminated with diesel and received six doses of P. CO2 emission was used to estimate biodegradation of hydrocarbons. Biodegradation increased with P doses. The P level that provided the highest hydrocarbon biodegradation rate showed linear and negative correlation with P-rem. The result shows that the requirement for P decreases as the P-rem of the soil increases (or the P-fixing capacity decreases). The dose of P recommended to maximize hydrocarbon biodegradation rate in soil can be estimated by the formula P (mg/dm3) = 436.5-5.39 × P-rem (mg/L).

Biocontrol capacity of bacteria isolated from sawdust of the dairy cattle production environment.

This research paper aimed to find endemic bacteria from the cattle production system to control the growth of mastitis pathogens. Bacteria were isolated from compost barn sawdust of two dairy cattle systems and later tested to verify their ability to control the growth of Staphylococcus aureus isolates obtained from cattle with mastitis. Bacterial isolates from these systems were tested to verify biocontrol capacity using the double-layer method. A total of 189 isolates were obtained from all samples by considering the morphology of the different bacterial colonies, with 30 isolates showing positive results for the growth control of at least one S. aureus strain and 19 isolates showing the ability to control more than one pathogen strain. The ability to control more than one pathogen and present a significant halo of inhibition in our isolates represents positive traits in the search for cattle mastitis biocontrol microorganisms. Thus, the results obtained represent the range of bacteria capable of controlling the pathogens without the use of antibiotics.

Prokaryotic community diversity during bioremediation of crude oil contaminated oilfield soil: effects of hydrocarbon concentration and salinity.

Crude oil extracted from oilfield reservoirs brings together hypersaline produced water. Failure in pipelines transporting this mixture causes contamination of the soil with oil and hypersaline water. Soil salinization is harmful to biological populations, impairing the biodegradation of contaminants. We simulated the contamination of a soil from an oilfield with produced water containing different concentrations of NaCl and crude oil, in order to evaluate the effect of salinity and hydrocarbon concentration on prokaryote community structure and biodegradation activity. Microcosms were incubated in CO2-measuring respirometer. After the incubation, residual aliphatic hydrocarbons were quantified and were performed 16S rRNA gene sequencing. An increase in CO2 emission and hydrocarbon biodegradation was observed with increasing oil concentration up to 100 g kg-1. Alpha diversity decreased in oil-contaminated soils with an increase in the relative abundance of Actinobacteria and reduction of Bacteroidetes with increasing oil concentration. In the NaCl-contaminated soils, alpha diversity, CO2 emission, and hydrocarbon biodegradation decreased with increasing NaCl concentration. There was an increase in the relative abundance of Firmicutes and Proteobacteria and a reduction of Actinobacteria with increasing salt concentration. Our results highlight the need to adopt specific bioremediation strategies in soils impacted by mixtures of crude oil and hypersaline produced water.

Rhamnolipids on Aedes aegypti larvae: a potential weapon against resistance selection.

The resistance of Aedes aegypti to chemical insecticides has been reported and our work proposes the use of biosurfactants as an alternative larvicide. We evaluated the effect of rhamnolipids against larvae of pyrethroid-resistant and susceptible A. aegypti strains. Time-mortality and sublethal effects were evaluated via survival analysis and swimming behavior, respectively. Rhamnolipids showed larvicidal effect at all tested concentrations. Rhamnolipids at 300 mg L-1 killed 100% of both susceptible and resistant larvae within 24 h of exposure and 99% after 30-days stored (pyrethroid-susceptible larvae). Regarding the sublethal effects, the swimming rate was reduced in 50 and 100 mg L-1 of rhamnolipids in grouped (pyrethroid-susceptible) larvae. Rhamnolipids at 50 mg L-1 reduced the distance and speed and increased the number of stops and resting time of individualized pyrethroid-susceptible larvae. The larvicidal effect of the rhamnolipids evaluated demonstrates that these compounds represent an alternative to control A. aegypti.

Hydrocarbonoclastic bacterial species growing on hexadecane: Implications for bioaugmentation in marine ecosystems.

LIMITATIONS: of bioaugmentation strategies are an obstacle for damage mitigation caused by oil spills in marine environments. Cells added to the contaminated sites are quickly lost by low adherence to the contaminants, rendering ineffective. This study used two hydrocarbonoclastic species - Rhodococcus rhodochrous TRN7 and Nocardia farcinica TRH1 cells - growing in mineral medium containing hexadecane to evaluate cell distribution in a crude-oil contaminated marine water. Cell affinity to hydrophobic compounds was quantified using Microbial Adhesion to Hydrocarbons test and analysis of fatty acids profile was performed using the Microbial Identification System. Bioremediation simulations were set up and cell populations of both strains were quantified by Fluorescent in situ Hybridization. R. rhodochrous and N. farcinica reached up to 97% and 60% of adhesion to hexadecane, respectively. The carbon source had more influence on the fatty acid profiles of both strains than the microbial species. The presence of 45.24% of 13:0 anteiso on total fatty acids in R. rhodochrous and 12.35% of saturated fatty acids with less than 13 carbons atoms in N. farcinica, as well as the occurrence of fatty alcohols only in presence of hexadecane in both species, are indicators that fatty acid changes are involved in the adaptation of the cells to remain at the water/oil interface. Cell quantification after bioremediation simulations revealed an increase in the density of both species, suggesting that the bioremediation strategies resulted on the increase of hydrocarbonoclastic species and up to 27.9% of all prokaryotic microbial populations in the microcosms were composed of R. rhodochrous or N. farcinica. These findings show the potential of application of these two bacterial strains in bioaugmentation of hydrocarbon-contaminated marine ecosystems.R. rhodochrous TRN7 and N. farcinica TRH1 hydrocarbonoclastic strains modify the fatty acid profile and increases density, optimizing hydrocarbons biodegradation.

Strategy to improve crude oil biodegradation in oligotrophic aquatic environments: W/O/W fertilized emulsions and hydrocarbonoclastic bacteria.

We studied petroleum biodegradation by biostimulation by using water in oil in water (W/O/W) double emulsions. These emulsions were developed using seawater, canola oil, surfactants, and mineral salts as sources of NPK. The emulsions were used in the simulation of hydrocarbon bioremediation in oligotrophic sea water. Hydrocarbon biodegradation was evaluated by CO2 emissions from microcosms. We also evaluated the release of inorganic nutrients and the stability of the emulsion's droplets. The double emulsions improved CO2 emission from the microcosms, suggesting the increase in the hydrocarbon biodegradation. Mineral nutrients were gradually released from the emulsions supporting the hydrocarbon biodegradation. This was attributed to the formation of different diameters of droplets and therefore, varying stabilities of the droplets. Addition of the selected hydrocarbonoclastic isolates simulating bioaugmentation improved the hydrocarbon biodegradation. We conclude that the nutrient-rich W/O/W emulsion developed in this study is an effective biostimulation agent for bioremediation in oligotrophic aquatic environments.

Changes in the microbial community during bioremediation of gasoline-contaminated soil.

We aimed to verify the changes in the microbial community during bioremediation of gasoline-contaminated soil. Microbial inoculants were produced from successive additions of gasoline to municipal solid waste compost (MSWC) previously fertilized with nitrogen-phosphorous. To obtain Inoculant A, fertilized MSWC was amended with gasoline every 3 days during 18 days. Inoculant B received the same application, but at every 6 days. Inoculant C included MSWC fertilized with N-P, but no gasoline. The inoculants were applied to gasoline-contaminated soil at 10, 30, or 50g/kg. Mineralization of gasoline hydrocarbons in soil was evaluated by respirometric analysis. The viability of the inoculants was evaluated after 103 days of storage under refrigeration or room temperature. The relative proportions of microbial groups in the inoculants and soil were evaluated by FAME. The dose of 50g/kg of inoculants A and B led to the largest CO2 emission from soil. CO2 emissions in treatments with inoculant C were inversely proportional to the dose of inoculant. Heterotrophic bacterial counts were greater in soil treated with inoculants A and B. The application of inoculants decreased the proportion of actinobacteria and increased of Gram-negative bacteria. Decline in the density of heterotrophic bacteria in inoculants occurred after storage. This reduction was bigger in inoculants stored at room temperature. The application of stored inoculants in gasoline-contaminated soil resulted in a CO2 emission twice bigger than that observed in uninoculated soil. We concluded that MSWC is an effective material for the production of microbial inoculants for the bioremediation of gasoline-contaminated soil.

Changes in the microbial community during bioremediation of gasoline-contaminated soil

Abstract We aimed to verify the changes in the microbial community during bioremediation of gasoline-contaminated soil. Microbial inoculants were produced from successive additions of gasoline to municipal solid waste compost (MSWC) previously fertilized with nitrogen-phosphorous. To obtain Inoculant A, fertilized MSWC was amended with gasoline every 3 days during 18 days. Inoculant B received the same application, but at every 6 days. Inoculant C included MSWC fertilized with N–P, but no gasoline. The inoculants were applied to gasoline-contaminated soil at 10, 30, or 50 g/kg. Mineralization of gasoline hydrocarbons in soil was evaluated by respirometric analysis. The viability of the inoculants was evaluated after 103 days of storage under refrigeration or room temperature. The relative proportions of microbial groups in the inoculants and soil were evaluated by FAME. The dose of 50 g/kg of inoculants A and B led to the largest CO2 emission from soil. CO2 emissions in treatments with inoculant C were inversely proportional to the dose of inoculant. Heterotrophic bacterial counts were greater in soil treated with inoculants A and B. The application of inoculants decreased the proportion of actinobacteria and increased of Gram-negative bacteria. Decline in the density of heterotrophic bacteria in inoculants occurred after storage. This reduction was bigger in inoculants stored at room temperature. The application of stored inoculants in gasoline-contaminated soil resulted in a CO2 emission twice bigger than that observed in uninoculated soil. We concluded that MSWC is an effective material for the production of microbial inoculants for the bioremediation of gasoline-contaminated soil.