Isolation and Selection of a Highly Tolerant Microbial Consortium with Potential for PAH Biodegradation from Heavy Crude Oil-Contaminated Soils

Zafra, German; Absalón, Ángel E.; Cuevas, Ma. Del Carmen; Cortés-Espinosa, Diana V.

doi:10.1007/s11270-013-1826-4

Cited by 101 publications

(49 citation statements)

References 54 publications

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“…The change in patterns of sporulation and pigmentation of fungi as a result of PAHs presence was also mentioned by Zafra et al (2014) and ReyesCésar, Absálon, Fernández, González, and Cortés-Espinosa (2014). The literature mentions that phenanthrene may inhibit spore germination of fungi (Lisowska, Palecz, & Dlugonski, 2004).…”

Section: Discussionmentioning

confidence: 54%

<b>Tolerance to Polycyclic Aromatic Hydrocarbons (PAHs) by filamentous fungi isolated from contaminated sediment in the Amazon region

Souza

Barreto

Mota

et al. 2017

Acta Sci. Biol. Sci.

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ABSTRACT. Tolerance to Polycyclic Hydrocarbons Aromatic (PAHs) is considered an important characteristic when assessing the bioremediation potential of microorganisms. Given this, the objective of this research was to assay filamentous fungi from the Amazon region, isolated from sediments with differents levels of contamination by PAHs, for tolerance to phenanthrene and pyrene. To achieve this, fungal cultures plugs (5 mm), obtained after 7 days growth, were transferred to petri dishes containing 20% Sabouraud dextrose agar medium, after surface innoculation with phenanthrene and pyrene crystals, separately. Radial mycelial growth was evaluated after 10 days at five different concentration levels for each contaminant and control group, all in triplicate for each treatment. Fungal growth and growth inhibition rates were calculated. The average growth of the colonies in each treatment was compared with one-way ANOVA, followed by a Tukey Test (p < 0,05). All fungi showed tolerant to phenanthrene and pyrene. However, Hypoxylon sp. showed the lowest growth inhibition rate and average growth rates significantly different of the other six tested species. Hypoxylon sp. has been shown to be a promising genetic resource for use in new studies of PAHs degradation.Keywords: phenanthrene, pyrene, fungal growth inhibition, Amazon.Tolerância a Hidrocarbonetos Policíclicos Aromáticos (HPAs) por fungos filamentosos isolados de sedimentos contaminados da região Amazônica RESUMO. A tolerância a Hidrocarbonetos Policíclicos Aromática (HPAs) é considerada como uma característica importante na avaliação do potencial de micro-organismos para biorremediação. Diante disso, o objetivo desta pesquisa foi avaliar fungos filamentosos da região amazônica, isolados de sedimentos com diferentes níveis de contaminação por HPAs, quanto à tolerância ao fenantreno e pireno. Para tanto, discos das culturas fúngicas (5 mm), obtidas após 7 dias de crescimento, foram transferidas para placas de Petri contendo meio Agar Sabouraud Dextrose a 20%, após inoculação superficial com cristais de fenantreno e pireno, separadamente. O crescimento micelial radial foi avaliado após 10 dias em cinco concentrações diferentes para cada contaminante e grupo controle, ambos em triplicata para cada tratamento. As taxas de crescimento fúngico e de inibição de crescimento foram calculadas. O crescimento médio das colônias em cada tratamento foi comparado com ANOVA one way, seguido pelo teste de Tukey (p < 0,05). Todos os fungos mostraram tolerância ao fenantreno e ao pireno. No entanto, Hypoxylon sp. apresentou menor taxa de inibição de crescimento e taxas médias de crescimento significativamente diferentes das outras seis espécies testadas. Hypoxylon sp. tem se mostrado um recurso genético promissor para uso em novos estudos sobre degradação de HPAs.Palavras-chaves: fenantreno, pireno, inibição do crescimento fúngico, Amazônia.

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Section: Discussionmentioning

confidence: 54%

<b>Tolerance to Polycyclic Aromatic Hydrocarbons (PAHs) by filamentous fungi isolated from contaminated sediment in the Amazon region

Souza

Barreto

Mota

et al. 2017

Acta Sci. Biol. Sci.

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“…Juhasz et al found that a Burkholderia cepacia strain isolated from soil near a manufacturing gas plant could degrade BaP with pyrene as the carbon source, although only 1.4 to 6.2% BaP was removed after 56 days (42). A Delftia strain was isolated from the microbial consortium of a crude oil-contaminated soil and removed 56.6% BaP in PAH-contaminated soil after 14 days (43). To our knowledge, prior to this study, BaP degradation by Burkholderia-related microorganisms using DNA-SIP had not been documented.…”

Section: Discussionmentioning

confidence: 99%

Identification of Benzo[ a ]pyrene-Metabolizing Bacteria in Forest Soils by Using DNA-Based Stable-Isotope Probing

Song

Luo

Jiang

et al. 2015

Appl Environ Microbiol

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c DNA-based stable-isotope probing (DNA-SIP) was used in this study to investigate the uncultivated bacteria with benzo[a]pyrene (BaP) metabolism capacities in two Chinese forest soils (Mt. Maoer in Heilongjiang Province and Mt. Baicaowa in Hubei Province). We characterized three different phylotypes with responsibility for BaP degradation, none of which were previously reported as BaP-degrading microorganisms by SIP. In Mt. Maoer soil microcosms, the putative BaP degraders were classified as belonging to the genus Terrimonas (family Chitinophagaceae, order Sphingobacteriales), whereas Burkholderia spp. were the key BaP degraders in Mt. Baicaowa soils. The addition of metabolic salicylate significantly increased BaP degradation efficiency in Mt. Maoer soils, and the BaP-metabolizing bacteria shifted to the microorganisms in the family Oxalobacteraceae (genus unclassified). Meanwhile, salicylate addition did not change either BaP degradation or putative BaP degraders in Mt. Baicaowa. Polycyclic aromatic hydrocarbon ring-hydroxylating dioxygenase (PAH-RHD) genes were amplified, sequenced, and quantified in the DNA-SIP 13 C heavy fraction to further confirm the BaP metabolism. By illuminating the microbial diversity and salicylate additive effects on BaP degradation across different soils, the results increased our understanding of BaP natural attenuation and provided a possible approach to enhance the bioremediation of BaP-contaminated soils. P olycyclic aromatic hydrocarbons (PAHs), a class of persistent organic pollutants (POPs), enter the environment through both natural and anthropogenic pathways. PAHs are released into the environment by means of natural processes, such as forest fires and direct biosynthesis under the action of microbes and plants (1). The primary artificial source of PAHs is the incomplete combustion of organic matter at high temperatures caused by human activities (2). Their presence in the environment poses a severe threat to public and ecosystem health because of their known acute toxicity and mutagenic, teratogenic, and carcinogenic features; they are therefore classified as priority pollutants by the U.S. Environmental Protection Agency (3). Furthermore, the persistence and genotoxicity of PAHs increase with molecular weight, and the presence of high-molecular-weight (HMW) PAHs in the environment is of greater concern. Benzo[a]pyrene (BaP), a representative HMW PAH with a five-ring structure, is a widespread pollutant with potent mutagenic and carcinogenic properties (4, 5). BaP is therefore identified as the first class of "human carcinogens" according to the report of the World Health Organization (WHO) International Agency for Research on Cancer (6). Generally, in soils with no industrial contamination, BaP concentrations vary from 3.5 to 3,700 g/kg, with a median concentration of 16 g/kg of soil; in contaminated soils and sediments, BaP concentrations range from 82 to 536 mg/kg (7).Bacteria possessing the ability of BaP utilization are readily isolated from contaminated soils or se...

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“…The three types of crude oil media were prepared by mixing crude oil (Gulf of Mexico, Montreal pipeline) to a final volume of 5% with cooling culture media containing either TSA, M9 plus glucose, or M9 alone. Crude oil was mixed 2:1 with acetone before addition to culture media in order to create a more homogeneous mixture, and acetone was allowed to volatilize under sterile conditions as was done by Zafra et al (29). The addition of crude oil to warm media may have sped up the volatilization of certain compounds (and may not perfectly reflect the crude oil added to soil microcosms), but likely most of those that are of interest in bioremediation (i.e., persistent compounds) were retained.…”

Section: Methodsmentioning

confidence: 99%

A Diverse Soil Microbiome Degrades More Crude Oil than Specialized Bacterial Assemblages Obtained in Culture

Bell

Stefani

Abram

et al. 2016

Appl Environ Microbiol

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Soil microbiome modification may alter system function, which may enhance processes like bioremediation. In this study, we filled microcosms with gamma-irradiated soil that was reinoculated with the initial soil or cultivated bacterial subsets obtained on regular media (REG-M) or media containing crude oil (CO-M). We allowed 8 weeks for microbiome stabilization, added crude oil and monoammonium phosphate, incubated the microcosms for another 6 weeks, and then measured the biodegradation of crude oil components, bacterial taxonomy, and functional gene composition. We hypothesized that the biodegradation of targeted crude oil components would be enhanced by limiting the microbial taxa competing for resources and by specifically selecting bacteria involved in crude oil biodegradation (i.e., CO-M). Postincubation, large differences in taxonomy and functional gene composition between the three microbiome types remained, indicating that purposeful soil microbiome structuring is feasible. Although phylum-level bacterial taxonomy was constrained, operational taxonomic unit composition varied between microbiome types. Contrary to our hypothesis, the biodegradation of C 10 to C 50 hydrocarbons was highest when the original microbiome was reinoculated, despite a higher relative abundance of alkane hydroxylase genes in the CO-M microbiomes and of carbon-processing genes in the REG-M microbiomes. Despite increases in the relative abundances of genes potentially linked to hydrocarbon processing in cultivated subsets of the microbiome, reinoculation of the initial microbiome led to maximum biodegradation. IMPORTANCEIn this study, we show that it is possible to sustainably modify microbial assemblages in soil. This has implications for biotechnology, as modification of gut microbial assemblages has led to improved treatments for diseases like Clostridium difficile infection. Although the soil environment determined which major phylogenetic groups of bacteria would dominate the assemblage, we saw differences at lower levels of taxonomy and in functional gene composition (e.g., genes related to hydrocarbon degradation). Further studies are needed to determine the success of such an approach in nonsterile environments. Although the biodegradation of certain crude oil fractions was still the highest when we inoculated with the diverse initial microbiome, the possibility of discovering and establishing microbiomes that are more efficient in crude oil degradation is not precluded. O il production, oil spills, and the storage of oily wastes by the petroleum industry have led to massive releases of petroleum hydrocarbons into the environment, making them some of the most ubiquitous environmental pollutants (1). Conventional decontamination technologies such as excavation (i.e., dig and dump), incineration, and chemical treatment of contaminants are costly and may further disrupt disturbed ecosystems (2, 3). When effective, bioremediation is a cheaper and more sustainable approach to decontamination. Bioaugmentation, the additio...

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Isolation and Selection of a Highly Tolerant Microbial Consortium with Potential for PAH Biodegradation from Heavy Crude Oil-Contaminated Soils

Cited by 101 publications

References 54 publications

<b>Tolerance to Polycyclic Aromatic Hydrocarbons (PAHs) by filamentous fungi isolated from contaminated sediment in the Amazon region

<b>Tolerance to Polycyclic Aromatic Hydrocarbons (PAHs) by filamentous fungi isolated from contaminated sediment in the Amazon region

Identification of Benzo[ a ]pyrene-Metabolizing Bacteria in Forest Soils by Using DNA-Based Stable-Isotope Probing

A Diverse Soil Microbiome Degrades More Crude Oil than Specialized Bacterial Assemblages Obtained in Culture

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