Advances in bacterial Rieske non-heme iron ring-hydroxylating dioxygenases that initiate polycyclic aromatic hydrocarbons degradation / 生物工程学报

Polycyclic aromatic hydrocarbons (PAHs) are a class of persistent organic pollutants, which have received widespread attentions due to their carcinogenic and mutagenic toxicity. The microbial degradation of PAHs are usually started from the hydroxylation, followed by dehydrogenation, ring cleavage and step-by-step removal of branched chains, and finally mineralized by the tricarboxylic acid cycle. Rieske type non-heme iron aromatic ring-hydroxylating dioxygenases (RHOs) or cytochrome P450 oxidases are responsible for the conversion of hydrophobic PAHs into hydrophilic derivatives by the ring hydroxylation. The ring hydroxylation is the first step of PAHs degradation and also one of the rate-limiting steps. Here, we review the distribution, substrate specificity, and substrate recognition mechanisms of RHOs, along with some techniques and methods used for the research of RHOs and PAHs.

Determination of Polycyclic Aromatic Hydrocarbons in Human Blood Samples Using Solid Phase Extraction and Gas Chromatography Mass Spectrometry- A Review

Abstract@#Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants with toxic effects and adverse health impacts on general population. Several methods of extraction had been applied to extract PAHs from human blood samples such as solid phase extraction (SPE). The SPE represents one of the most common techniques for extraction and clean-up procedures as it needs low quantity of solvents with less manual efforts. Similarly, various analytical instruments like gas chromatography coupled to mass spectrometry (GC-MS) was used to measure the PAHs levels. Gas chromatography is a simple, fast, and very efficient method for solvents and small organic molecules. This review provides an overview of the measured concentrations of PAHs in human blood samples through the application of SPE and GCMS during the last ten years. While these studies used various solvents, their application of SPE method and GC-MS revealed rewarding results about the determination of PAHs levels in the human samples.

Induced CYP1A activity and DNA damage in fish from the middle Paraíba do Sul River basin, southeastern Brazil / Atividade induzida de CYP1A e danos de DNA em peixes do trecho médio da bacia do rio Paraíba do Sul, sudeste do Brasil

The Paraíba do Sul River (PSR) drainage basin in Southeastern Brazil covers one of the most industrialized and densely populated regions of the country. The impact of chemical contamination on the PSR basin seems to be more pronounced in its middle reach where a number of potentially polluting plants are located. In this study, we used hepatic EROD activity - a biomarker of exposure to CYP1A-inducing pollutants (e.g. PAHs, PCDD/Fs, PCBs) - and the incidence of micronucleated erythrocytes (Mn) in the peripheral blood - a biomarker of effects of DNA-damaging agents (e.g. PAHs) - to evaluate the effects of pollution on two native fish species, Geophagus brasiliensis and Pimelodus maculatus. Results showed that the incidence of Mn and EROD in G. brasiliensis and P. maculatus from the two most downstream sites (Três Rios Town and Piabanha River) were markedly higher than the incidence of Mn and EROD recorded in fish from the most upstream site (Funil Reservoir). Our findings are consistent with the view that CYP1A-inducing activity and increased DNA-damage are found in fish caught in sampling sites located downstream of the stretch where there are several industries that are potential sources of PAHs and CYP1A-inducing contaminants.

O rio Paraíba do Sul (RPS) drena uma das mais industrializadas e densamente povoadas áreas do sudeste do Brasil. O impacto de contaminação química no RPS parece ser mais pronunciado no segmento médio da bacia onde se localiza grande número de indústrias potencialmente poluidoras. Neste estudo, foi avaliada a atividade hepática EROD – um biomarcador de exposição a poluente indutores da CYP1A (e.g. HAPs, PCDD/Fs, PCBs) – e a incidência de eritrócitos micronucleados (Mn) no sangue periférico – um biomarcador de efeitos de agentes de danificação do DNA- (e.g. HAPs) – para avaliar o efeito de poluição sobre dois peixes nativos, Geophagus brasiliensis e Pimelodus maculatus. Os resultados apresentaram que a incidência de Mn e EROD em G. brasiliensis e P. maculatus nos dois locais no trecho mais baixo (Três Rios e rio Piabanha) foi marcadamente mais elevada do que a incidência e a atividade EROD registradas em peixes dos locais mais à montante (reservatório do Funil). Estes resultados são consistentes com a visão de que atividade indutora de CYP1A e elevadora de danos no DNA são encontradas em peixes capturados em locais abaixo do trecho onde grande número de indústrias são potenciais fontes de poluição de indutores de contaminantes HAPs e CYP1A.

Bioremediation of polycyclic aromatic hydrocarbon (PAH) compounds: (acenaphthene and fluorene) in water using indigenous bacterial species isolated from the Diep and Plankenburg rivers, Western Cape, South Africa

Abstract This study was conducted to investigate the occurrence of PAH degrading microorganisms in two river systems in the Western Cape, South Africa and their ability to degrade two PAH compounds: acenaphthene and fluorene. A total of 19 bacterial isolates were obtained from the Diep and Plankenburg rivers among which four were identified as acenaphthene and fluorene degrading isolates. In simulated batch scale experiments, the optimum temperature for efficient degradation of both compounds was determined in a shaking incubator after 14 days, testing at 25 °C, 30 °C, 35 °C, 37 °C, 38 °C, 40 °C and 45 °C followed by experiments in a Stirred Tank Bioreactor using optimum temperature profiles from the batch experiment results. All experiments were run without the addition of supplements, bulking agents, biosurfactants or any other form of biostimulants. Results showed that Raoultella ornithinolytica, Serratia marcescens, Bacillus megaterium and Aeromonas hydrophila efficiently degraded both compounds at 37 °C, 37 °C, 30 °C and 35 °C respectively. The degradation of fluorene was more efficient and rapid compared to that of acenaphthene and degradation at Stirred Tank Bioreactor scale was more efficient for all treatments. Raoultella ornithinolytica, Serratia marcescens, Bacillus megaterium and Aeromonas hydrophila degraded a mean total of 98.60%, 95.70%, 90.20% and 99.90% acenaphthene, respectively and 99.90%, 97.90%, 98.40% and 99.50% fluorene, respectively. The PAH degrading microorganisms isolated during this study significantly reduced the concentrations of acenaphthene and fluorene and may be used on a larger, commercial scale to bioremediate PAH contaminated river systems.

Sphingomonas from petroleum-contaminated soils in Shenfu, China and their PAHs degradation abilities

Abstract Members of the Sphingomonas genus are often isolated from petroleum-contaminated soils due to their unique abilities to degrade polycyclic aromatic hydrocarbons (PAHs), which are important for in situ bioremediation. In this study, a combined phenotypic and genotypic approach using streptomycin-containing medium and Sphingomonas -specific PCR was developed to isolate and identify culturable Sphingomonas strains present in petroleum-contaminated soils in the Shenfu wastewater irrigation zone. Of the 15 soil samples examined, 12 soils yielded yellow streptomycin-resistant colonies. The largest number of yellow colony-forming units (CFUs) could reach 105 CFUs g-1 soil. The number of yellow CFUs had a significant positive correlation (p < 0.05) with the ratio of PAHs to total petroleum hydrocarbons (TPH), indicating that Sphingomonas may play a key role in degrading the PAH fraction of the petroleum contaminants at this site. Sixty yellow colonies were selected randomly and analyzed by colony PCR using Sphingomonas -specific primers, out of which 48 isolates had PCR-positive signals. The 48 positive amplicons generated 8 distinct restriction fragment length polymorphism (RFLP) patterns, and 7 out of 8 phylotypes were identified as Sphingomonas by 16S rRNA gene sequencing of the representative strains. Within these 7 Sphingomonas strains, 6 strains were capable of using fluorene as the sole carbon source, while 2 strains were phenanthrene-degrading Sphingomonas. To the best of our knowledge, this is the first report to evaluate the relationship between PAHs contamination levels and culturable Sphingomonas in environmental samples.

Morphological changes and growth of filamentous fungi in the presence of high concentrations of PAHs

In this study, we evaluated the effect of low and high molecular weight polycyclic aromatic hydrocarbons (PAHs), i.e., Phenanthrene, Pyrene and Benzo[a]pyrene, on the radial growth and morphology of the PAH-degrading fungal strains Aspergillus nomius H7 and Trichoderma asperellum H15. The presence of PAHs in solid medium produced significant detrimental effects on the radial growth of A. nomius H7 at 4,000 and 6,000 mg L⁻¹ and changes in mycelium pigmentation, abundance and sporulation ability at 1,000–6,000 mg L⁻¹. In contrast, the radial growth of T. asperellum H15 was not affected at any of the doses tested, although sporulation was observed only up to 4,000 mg L⁻¹ and as with the H7 strain, some visible changes in sporulation patterns and mycelium pigmentation were observed. Our results suggest that fungal strains exposed to high doses of PAHs significantly vary in their growth rates and sporulation characteristics in response to the physiological and defense mechanisms that affect both pigment production and conidiation processes. This finding is relevant for obtaining a better understanding of fungal adaptation in PAH-polluted environments and for developing and implementing adequate strategies for the remediation of contaminated soils.

Relationship between PAHs Concentrations in Ambient Air and Deposited on Pine Needles

OBJECTIVES: This study was carried out to determine whether or not pine needles can be used as passive samplers of atmospheric polycyclic aromatic hydrocarbons (PAHs) using the correlation between accumulated PAH concentrations in air (Ca, ng/m3) and those deposited on pine needles (Cp, ng/g dry). METHODS: PAHs in ambient air was collected using low volume PUF sampler and pine needles was gathered at same place for 7 months. RESULTS: good correlation (R2=0.8582, p<0.05) was found between Ca and Cp for PAHs with a higher gaseous state in air (AcPy, Acp, Flu, Phen, Ant, Flt, Pyr, BaA and Chry), but there was a poorer correlation (R2=0.1491, p=0.5123) for the PAHs with a lower gaseous state (BbF, BkF, BaP, DahA, BghiP and Ind123). A positive correlation (R2=0.8542) was revealed between the logarithm of the octanol-air partitioning coefficient (logKoa) and Cp/Ca for the PAHs with a higher gaseous state in air, but there was a negative correlation (R2=0.8131) for the PAHs with a lower gaseous state. The Ca-Cp model could not be used to estimate PAHs concentrations in air using deposited PAHs concentrations on pine needles, but the logKoa-Cp/Ca model could be used. CONCLUSIONS: It was found that pine needles can be used as passive samplers of atmospheric PAHs.

Application of molecular fingerprinting for analysis of a PAH-contaminated soil microbiota growing in the presence of complex PAHs / Aplicação de técnica molecular para análise da microbiota de solo contaminado em misturas complexas de HPAs

Polycyclic aromatic hydrocarbons (PAHs) constitute a group of priority pollutants which are present at high concentrations in the soils of many industrial contaminated sites. Pollution by these compounds may stimulate growth of organisms able to live in these environments causing changes in the structure of the microbial community due to some cooperative process of metabolization of toxic compounds. A long-term PAH-contaminated soil was stored for several years and used to analyze the native microbiota regarding their ability to grow on pyrene, benzo[a]pyrene, as well as in mixtures of LMW- and HMW-PAHs. Molecular profiles of the microbial community was assessed by PCR-DGGE of 16S rRNA gene, and the number of bands observed in DGGE analyses was interpreted as dominant microbial members into the bacterial community. Results of PAH-contaminated soil microorganisms showed different profiles in the degradative dynamics when some nutrients were added. Predominant species may play a significative role while growing and surviving on PAHs, and some other metabolically active species have emerged to interact themselves in a cooperative catabolism of PAHs.

Os hidrocarbonetos poliaromáticos (HPAs) são considerados poluentes prioritários presentes em expressiva concentração no solo contaminado com derivados de petróleo. A poluição por esses compostos estimula o crescimento de microrganismos capazes de sobreviverem nestes ambientes contaminados, causando alterações na estrutura da comunidade microbiana do solo pelo processo de cooperação metabólica entre as populações. Um solo contaminado por um longo período de tempo foi coletado de uma área industrial (Port Melbourne, Austrália) e utilizado para análise da capacidade da comunidade microbiana em crescer em HPAs isolados e/ou em misturas como únicas fontes de carbono e energia. Os perfis moleculares foram obtidos por PCR-DGGE do fragmento da subunidade 16S do DNA ribossomal, sendo o número de bandas presentes nos géis de eletroforese interpretado como populações dominantes dentro da comunidade do solo. Resultados demonstraram diferentes perfis quando vitaminas e micronutrientes fizeram parte do meio líquido de crescimento da microbiota, além da presença dos HPAs. As espécies predominantes podem ter papel significativo na degradação desses compostos no solo a ser remediado, enquanto sobrevivem e interagem com outras populações por meio de mecanismos de catabolismo cooperativo.