Yujiao Sun

Whole cell bioreporter application for rapid detection and evaluation of crude oil spill in seawater caused by Dalian oil tank explosion.

Accidents involving the release of crude oil to seawater pose serious threat to human and animal health, fisheries and marine ecosystems. A whole cell bioreporter detection method, which has unique advantages for the rapid evaluation on toxicity and bioavailability, is a useful tool to provide environmental risk assessments at crude oil-contaminated sites. Acinetobacter baylyi ADPWH_alk and ADPWH_recA are chromosomally-based alkane and genotoxicity bioreporters which can be activated to express bioluminescence in the presence of alkanes and genotoxic compounds. In this study, we applied Acinetobacter ADPWH_alk and ADPWH_recA bioreporters to examine six seawater and six sediment samples around the Dalian Bay four weeks after an oil tank explosion in Dalian, China in 2010, and compared the results with samples from the same sites one year after. The results of bioreporter detection suggest that seawater and sediments from five sites (DB, NT, JSB, XHP and FJZ) four weeks after the oil-spill were contaminated by the crude oil with various extents of genotoxicity. Among these six sites, DB and NT had high oil contents and genotoxicity, and JSB had high oil content but low genotoxicity in comparison with an uncontaminated site LSF, which is located at other side of the peninsula. These three sites (DB, NT and JSB) with detectable genotoxicity are within 30 km away from the oil spill point. The far-away two sites XHP (38.1 km) and FJZ (31.1 km) were lightly contaminated with oil but no genotoxicity suggesting that they are around the contamination boundary. Bioreporter detection also indicates that all six sites were clean one year after the oil-spill as the alkane and genotoxicity were below detection limit. This study demonstrates that bioreporter detection can be used as a rapid method to estimate the scale of a crude oil spill accident and to evaluate bioavailability and genotoxicity of contaminated seawater and sediments, which are crucial to risk assessment and strategic decision-making for environmental management and clean-up.

Evaluation on factors influencing the heterotrophic growth on the soluble microbial products of autotrophs

In this work, the heterotrophic growth on the microbial products of autotrophs and the effecting factors were evaluated with both experimental and modeling approaches. Fluorescence in situ hybridization (FISH) analysis illustrated that ammonia oxidizers (AOB), nitrite oxidizers (NOB), and heterotrophs accounted for about 65%, 20%, and 15% of the total bacteria, respectively. The mathematical evaluation of experimental data reported in literature indicated that heterotrophic growth in nitrifying biofilm (30–50%) and granules (30%) was significantly higher than that of nitrifying sludge (15%). It was found that low influent ammonium resulted in a lower availability of soluble microbial products (SMP) and a slower heterotrophic growth, but high ammonium (>150 mg N L−1) feeding would lead to purely AOB dominated sludge with high biomass‐associated products contained effluent, although the absolute heterotrophic growth increased. Meanwhile, the total active biomass concentration increased gradually with the increasing solids retention time, whereas the factions of active AOB, NOB, and heterotrophs varied a lot at different solids retention times. This work could be useful for better understanding of the autotrophic wastewater treatment systems. Biotechnol. Bioeng. 2011; 108:804–812.

Sequencing Insights into Microbial Communities in the Water and Sediments of Fenghe River, China

The connection between microbial community structure and spatial variation and pollution in river waters has been widely investigated. However, water and sediments together have rarely been explored. In this study, Illumina high-throughput sequencing was performed to analyze microbes in 24 water and sediment samples from natural to anthropogenic sources and from headstream to downstream areas. These data were used to assess variability in microbial community structure and diversity along in the Fenghe River, China. The relationship between bacterial diversity and environmental parameters was statistically analyzed. An average of 1682 operational taxonomic units was obtained. Microbial diversity increased from the headstream to downstream and tended to be greater in sediment compared with water. The water samples near the headstream endured relatively low Shannon and Chao1 indices. These diversity indices and the number of observed species in the water and sediment samples increase downstream. The parameters also differ in the two river tributaries. Community structures shift based on the extent of nitrogen pollution variation in the sediment and water samples. The four most dominant genera in the water community were Escherichia, Acinetobacter, Comamonadaceae, and Pseudomonas. In the sediments, the most dominant genera were Stramenopiles, Flavobacterium, Pseudomonas, and Comamonadaceae. The number of ammonia-oxidizing archaea in the headstream water slightly differed from that in the sediment but varied considerably in the downstream sediments. Statistical analysis showed that community variation is correlated with changes in ammonia nitrogen, total nitrogen, and nitrate nitrogen. This study identified different microbial community structures in river water and sediments. Overall this study emphasized the need to elucidate spatial variations in bacterial diversity in water and sediments associated with physicochemical gradients and to show the effects of such variation on waterborne microbial community structures.

Separating and characterizing functional alkane degraders from crude-oil-contaminated sites via magnetic nanoparticle-mediated isolation

Uncultivable microorganisms account for over 99% of all species on the planet, but their functions are yet not well characterized. Though many cultivable degraders for n-alkanes have been intensively investigated, the roles of functional n-alkane degraders remain hidden in the natural environment. This study introduces the novel magnetic nanoparticle-mediated isolation (MMI) technology in Nigerian soils and successfully separates functional microbes belonging to the families Oxalobacteraceae and Moraxellaceae, which are dominant and responsible for alkane metabolism in situ. The alkR-type n-alkane monooxygenase genes, instead of alkA- or alkP-type, were the key functional genes involved in the n-alkane degradation process. Further physiological investigation via a BIOLOG PM plate revealed some carbon (Tween 20, Tween 40 and Tween 80) and nitrogen (tyramine, l-glutamine and d-aspartic acid) sources promoting microbial respiration and n-alkane degradation. With further addition of promoter carbon or nitrogen sources, the separated functional alkane degraders significantly improved n-alkane biodegradation rates. This suggests that MMI is a promising technology for separating functional microbes from complex microbiota, with deeper insight into their ecological functions and influencing factors. The technique also broadens the application of the BIOLOG PM plate for physiological research on functional yet uncultivable microorganisms.

Characterizing Microbial Activity and Diversity of Hydrocarbon-Contaminated Sites

Hydrocarbons, one of the major petroleum constituents, mainly include saturated alkanes and cycloalkanes, unsaturated alkenes, alkynes and aromatic hydrocarbons. The usual composition of light crude oil is 78% saturates, 18% aromatics, 4% resins and <2% asphaltenes (Olah and Molnar 2003). The hydrocarbon fractions in the order of decreasing volatility are C6C10; C10 – C16; C16 C34 and C34 C50, in which Cx is referred to the number of carbon molecules in the alkane backbone.

Impacts of n-alkane concentration on soil bacterial community structure and alkane monooxygenase genes abundance during bioremediation processes

Petroleum hydrocarbons, mainly consisting of n-alkanes and polycyclic aromatic hydrocarbons (PAHs), are considered as priority pollutants and biohazards in the environment, eventually affecting the ecosystem and human health. Though many previous studies have investigated the change of bacterial community and alkane degraders during the degradation of petroleum hydrocarbons, there is still lack of understanding on the impacts of soil alkane contamination level. In the present study, microcosms with different n-alkane contamination (1%, 3% and 5%) were set up and our results indicated a complete alkane degradation after 30 and 50 days in 1%- and 3%-alkane treatments, respectively. In all the treatments, alkanes with medium-chain length (C11-C14) were preferentially degraded by soil microbes, followed by C27-alkane in 3% and 5% treatments. Alkane contamination level slightly altered soil bacterial community, and the main change was the presence and abundance of dominant alkane degraders. Thermogemmatisporaceae, Gemmataceae and Thermodesulfovibrionaceae were highly related to the degradation of C14- and C27-alkanes in 5% treatment, but linked to alkanes with medium-chain (C11-C18) in 1% treatment and C21-alkane in 3% treatment, respectively. Additionally, we compared the abundance of three alkane-monooxygenase genes, e.g., alk_A, alk_P and alk_R. The abundance of alk_R gene was highest in soils, and alk_P gene was more correlated with alkane degradation efficiency, especially in 5% treatment. Our results suggested that alkane contamination level showed non-negligible effects on soil bacterial communities to some extents, and particularly shaped alkane degraders and degrading genes significantly. This study provides a better understanding on the response of alkane degraders and bacterial communities to soil alkane concentrations, which affects their biodegradation process.

Methanogenic community structure in simultaneous methanogenesis and denitrification granular sludge

A laboratory scale up-flow anaerobic sludge bed (UASB) bioreactor fed with synthetic wastewater was operated with simultaneous methanogenesis and denitrification (SMD) granules for 235 days with a gradient decrease of C/N. Molecular cloning, qRT-PCR and T-RFLP were applied to study the methanogenic community structures in SMD granules and their changes in response to changing influent C/N. The results indicate that when C/N was 20:1, the methane production rate was fastest, and Methanosaetaceae and Methanobacteriaceae were the primary methanogens within the Archaea. The richness and evenness of methanogenic bacteria was best with the highest T-RFLP diversity index of 1.627 in the six granular sludge samples. When C/N was reduced from 20:1 to 5:1, the methanogenic activity of SMD granules decreased gradually, and the relative quantities of methanogens decreased from 36.5% to 10.9%. The abundance of Methanosaetaceae in Archaea increased from 64.5% to 84.2%, while that of Methanobacteriaceae decreased from 18.6% to 11.8%, and the richness and evenness of methanogens decreased along with the T-RFLP diversity index to 1.155, suggesting that the community structure reflected the succession to an unstable condition represented by high nitrate concentrations.

Research on Remediation of Petroleum Contaminated Soil by Plant-Inoculation Cold-Adapt Bacteria

With the rapid development of the petroleum industry, a large number of oil contaminants leaked into the soil in the petrochemical complex areas, gas stations, automobile factory and other places, which resulted in serious soil contamination. Micro-Biological degradation of oil pollution was the focus of the present study. Many hydrocarbon-contaminated environments were characterized by low or elevated temperatures, acidic or alkaline pH, high salt concentrations, or high pressure. Hydrocarbon-degrading microorganisms, adapted to grow and thrive in these environments, played an important role in the biological treatment of polluted extreme habitats.

Microcosm Study on Response of Soil Microorganisms to Polycyclic Aromatic Hydrocarbons Contamination

Soil and groundwater contamination by petroleum and coal tar becomes one of the urgent environmental issues concerned by more and more scientists with regards to the public health. With the induction of organic contaminants to soil and groundwater it will change the environmental conditions within the plume and hence lead to the variation of indigenous microorganisms. The article aims to investigate the response of indigenous microorganisms to the petroleum contamination. Three polycyclic aromatic hydrocarbons (PAHs) phenanthrene, anthracene and pyrene were selected as the contaminants and tested using laboratory microcosm. The response of indigenous microorganisms to the PAHs induction was analyzed by measuring the change of soil microbial activity, soil catalase activity, contaminants concentration and geochemical parameters. The experimental results showed that: (1) the major parts of the PAHs were absorbed by the soil, its transport was retarded, (2) the soil microbial activity was inhibited at the initial stage of PAHs induction and then slowly increased, (3) the soil catalase activity was stimulated at first with PAHs induction and then inhibited within the range -0.05 mL (0.1 M KMnO 4 )/g (soil).