Population structures shift during the biodegradation of crude and fuel oil by an indigenous consortium

Petroleum and fuel oil are complex mixtures of recalcitrant hydrocarbons. The biodegradation of these hydrocarbons needs the action of a vast variety of enzymatic capacities. A microbial consortium offers the capability to degrade complex substrates through the assembly of different biochemical reactions, providing a metabolic versatility superior to axenic cultures. In this work, the microbial population dynamics, taxonomy, and the catabolic capacity of a stabilized consortium exposed to fuel and crude oil was analyzed through metagenomics. The stabilized consortium degraded 59% of crude oil components after 8 days, and 34% of fuel oil components after 130 days. Population dynamics analysis indicates that in fuel oil the biodiversity richness was higher; however, denaturing gradient gel electrophoresis similarity dendrogram shows significant changes in the microbial population during crude oil degradation. Taxonomy studies indicate a great genera divergence; only eight microbial genera were common in both samples. In crude oil, the Limnobacter sp. was the most abundant specie (15.6%), while Sphingomonas wittichii (7.9%) and Novosphingobium aromaticivorans (7.6%) were abundant in fuel oil. These microorganisms have been reported to participate in the degradation of aliphatic and aromatic hydrocarbons. Functional analysis suggests that fuel and crude oil components changed the interactions between the consortium members affecting the collective metabolic functionality.     

Worldwide distribution and significance of secondary microbial methane formed during petroleum biodegradation in conventional reservoirs

Around half of world’s endowment of in-place oil and bitumen experienced biodegradation, which is now believed to be largely an anaerobic methanogenic process. However, the distribution and scale of methanogenic biodegradation in the world’s petroleum accumulations and the significance of its terminal product, secondary microbial methane, in the global gas endowment and carbon cycle are largely unknown. Here, I present geological and geochemical criteria to recognize secondary microbial methane in conventional petroleum reservoirs. These include the presence of biodegraded oil (as pools, legs or shows) in the reservoir or down-dip, the relatively dry (methane dominated) gas containing methane with δ¹³C values between −55‰ and −35‰ and, most importantly, CO₂ with δ¹³C > +2‰. Based on these criteria, the presence of secondary microbial methane is apparent in 22 basins, probable in 12 basins and possible in six basins worldwide. Reservoirs apparently containing secondary microbial methane are mostly Cenozoic and clastic and occur at depths of 37-1834 m below surface/mudline and temperatures of 12-71 °C. Using the current global endowment of in-place oil and bitumen and reasonable assumptions about conversion of oil into methane during biodegradation, I estimated that ∼65,500 tcf of secondary microbial methane could have been generated in existing worldwide accumulations of oil and bitumen through their geological history. From 1461-2760 tcf in-place (845-1644 tcf recoverable) of secondary microbial methane may be accumulated as free and oil-dissolved gas in petroleum reservoirs. I also updated the inventory of primary microbial methane and estimated that the global primary microbial gas endowment (free and oil-dissolved) is from 676-797 tcf in-place (407-589 tcf recoverable). Secondary microbial methane may account for ∼5-11% of the global conventional recoverable gas endowment and appears more abundant than primary microbial gas (∼3-4% of the global gas endowment). Most of the generated secondary microbial methane probably is aerobically and anaerobically oxidized to CO₂ in the overburden above petroleum reservoirs. However, some secondary microbial methane may escape from shallow reservoirs into the atmosphere and affect present and past global climate.     

Study of the biodegradation levels of oils from the Orinoco Oil Belt (Junin area) using different biodegradation scales

This work presents a study of the molecular composition of the saturated and aromatic hydrocarbon fractions of crude oils from the Orinoco Oil Belt (Junín area) in the Eastern Venezuelan Basin, with the purpose of classifying these samples following two distinct biodegradation assessment schemes: the PM scale [Peters, K.E., Moldowan, J.M., 1993. The Biomarker Guide: Interpreting Molecular Fossils in Petroleum and Ancient Sediments. Prentice Hall, Englewood Cliffs, New Jersey, p. 363] and the Manco scale [Larter, S., Huang, H., Adams, J., Bennett, B., Snowdon, L.R., 2012. A practical biodegradation scale for use in reservoir geochemical studies of biodegraded oils. Organic Geochemistry 45, 66–76]. Both scales agree on the presence of different levels of biodegradation for the analyzed oils, although they are based on different groups of compounds. The PM scale uses mainly compounds from the saturated hydrocarbon fractions (e.g., n-alkanes, acyclic isoprenoids, terpanes and steranes) as well as aromatic steroids. On the other hand, the Manco scale considers other compounds (e.g., alkyltoluenes, naphthalene, methylnaphthalene, phenanthrene, alkylphenanthrenes and methyldibenzothiophenes) not included in the PM biodegradation scale. Thus, the combined use of these two scales allows the determination of the level of biodegradation of both saturated and aromatic compound classes. Dibenzothiophene (DBT), which was not included for the Manco score determination, also shows variations in peak intensity when compared to C₄-alkylnaphthalenes, presumably associated with the process of biological alteration. The differences in the biodegradation levels observed in the present study may be attributed to variations in parameters that control biodegradation rates laterally across the study area or the existence of varying communities of microorganisms, among other possible factors.     

辽河原油好氧生物降解模拟过程中化学组成及其碳同位素值的变化

对辽河盆地中央凸起南部潜山原油进行了好氧生物降解模拟实验,分析了辽河原油在轻度至中度(PM 1~4级)的好氧生物降解过程中化学组分的变化,以及在此过程中原油族组分和正构烷烃单体的稳定碳同位素值的变化。结果显示,饱和烃组分的消耗速率最快,正构烷烃的消耗速率高于异构烷烃,低碳数正构烷烃的消耗速率高于高碳数正构烷烃。正构烷烃的疏水性及其在原油中的相对浓度可能是影响微生物选择性地消耗正构烷烃的主导因素,即低碳数的正构烷烃由于在油水界面上的表面张力较小而更容易被微生物降解。正构烷烃单体(C_(14)~C_(30))稳定碳同位素值在生物降解过程中较稳定,未产生明显的碳同位素分馏效应。正构烷烃的好氧降解产物主要是脂肪酸、醇和CO_2。辽河原油中正构烷烃组分的δ~(13)C值较饱和烃、芳烃、胶质和沥青质组分都要偏重,降解产物中水溶性较低的高碳数脂肪酸和醇大部分进入胶质中,少部分通过氢键缔合和形成酯键等方式进入到沥青质组分中,造成沥青质组分的δ~(13)C值发生相应的变化,即对沥青质的δ~(13)C值产生"拉动效应";而降解产物中高水溶性的低碳数烷基酸和醇会进入水相而脱离原油,导致残余油的整体δ~(13)C值相应变轻。因此,假如易受生物降解的烃类如正构烷烃等的δ~(13)C值与原油的δ~(13)C值差别比较大,那么生物降解过程中随着原油化学组成的改变,原油本身及其族组分的δ~(13)C值均可能发生明显的变化。     

Sorption and anaerobic biodegradation of soluble aromatic compounds during groundwater transport. 2. Computer modeling

. The natural attenuation of a binary mixture of toluene and 1,2,4-trimethylbenzene, which was previously experimentally demonstrated in anaerobic column experiments, was simulated with the newly-developed PHREEQC model to reveal the biogeochemical processes involved. The processes considered in this model include advection, diffusion, sorption, redox reactions (e.g., oxidation of toluene and 1,2,4-trimethylbenzene coupled to the reduction of microbial ferric iron), precipitation of secondary minerals such as siderite and magnetite, and dissolution of hydrous ferric oxides, as well as other thermodynamic equilibrium reactions. The functions of advection and diffusion built into the PHREEQC were conventionally simulated. The sorption was kinetically simulated based on a bicontinuum model, whereas biodegradation was described using a multiple-term Monod model. The processes mentioned above were coupled with a geochemical equilibrium model. The consistency between the experimental and modeling results indicates that the processes defined and the controlling parameters chosen could describe the coupled interactions between transport and biogeochemical reactions for the studied system. The discrepancy between modeling and experimental observations, however, also revealed the limitation of the modeling approach. In general, the model approach, which incorporates simplified mathematical representations of the true physico-chemical dynamics of the sorption and biodegradation processes of toluene and 1,2,4-trimethylbenzene under anaerobic conditions is a very efficient and good predictive tool in management and remediation of contaminant groundwater aquifers.     

Measurement of partition coefficients of phenol and cresols in gas-charged crude oil/water systems

An instrument has been constructed for monitoring the partition coefficients of phenol and cresols between crude oil and water under sub-surface conditions. The device has the capacity for introducing methane gas into crude oil, thereby allowing measurements under live oil (solution gas-containing) conditions. The partition coefficients of phenol and cresols have been measured in crude oil: water substrates under “live” oil and “dead” oil (without solution gas) conditions over a temperature range 25–150 °C. Over the range investigated it is seen that the introduction of gas (crude oil saturated at 100 bar with methane) into the system resulted in an approximate doubling of partition coefficients compared to the equivalent dead oil: brine systems. The partition coefficient data obtained using the device may be employed in a number of petroleum exploration and production activities such as the determination of residual oil saturation of a water-flooded petroleum reservoir. Partition coefficient measurements may help in predicting toxic organic solute loadings in oilfield discharge waters.     

Sorption and anaerobic biodegradation of soluble aromatic compounds during groundwater transport. 1. Laboratory column experiments

. This paper deals with sorption and anaerobic biodegradation of the soluble aromatic fraction of jet fuel and how it is influenced by pore-water velocity during transport in a groundwater aquifer. The study was carried out as controlled laboratory column experiments. A binary mixture of toluene and 1,2,4-trimethylbenzene with a concentration ratio of 2:1 was used through the entire investigations. The column experiments were conducted with contaminated sediments and groundwater, taken from wells at a field research site. The columns were operated anaerobically under continuous-flow conditions at 10 °C in a temperature-controlled refrigerator. Two percent sodium azide was added to the injection solution of two of the columns to prevent biodegradation of the studied organic mixture. Chloride was used as a conservative tracer to characterize the hydrodynamic parameters such as dispersivity and porosity of the columns. The results showed that both compounds in the mixture were attenuated because of sorption and biodegradation processes in the columns. 1,2,4-trimethylbenzene was attenuated more significantly than toluene. Biodegradation of toluene was coupled mainly with the microbial reduction of ferric iron, whereas 1,2,4-trimethylbenzene, in contrast, was mostly sorbed. Their sorption and biodegradation were studied with different pore-water velocities, and a mass balance approach was applied to calculate biodegradation rates. The biodegradation rates of toluene were –0.16, –0.21, and –0.26 (unit: mM day^–1) for pore-water velocities of 96, 82.4, and 54.9 (unit: cm day^–1), respectively. This indicates that a decrease in the pore-water velocity significantly enhanced the biodegradation of toluene, consistent with other reports in the literature. For 1,2,4-trimethylbenzene the biodegradation rates were –0.05, –0.13 (unit: mM day^–1) for pore-water velocities of 96 and 82.4 (unit: cm day^–1), respectively. The biodegradation rate of 1,2,4-trimethylbenzene did not increase at the lowest pore water velocity as expected. This might be a result of substrate competition.     

Effects of Biodegradation on the Distribution of Alkylcarbazoles in Crude Oils

We have investigated the distributions of alkylcarbazoles in a series of crude oils with different biodegradation extents, in combination with biomarker parameters, stable carbon isotopic ratios and viscosities. The analyses showed that slight biodegradation has little effect on alkylcarbazoles. The concentrations of C₀-, C₁-, and C₂-carbazoles seem to display a slight decrease with biodegradation through the moderately biodegraded stage, and an abrupt decrease to the heavily biodegraded stage. The relative concentrations of C₀-, C₁-, and C₂-carbazoles do not show any apparent change in the non-heavily biodegraded stages, but through non-heavily biodegraded to heavily biodegraded stages, the percentages of C₀- and C₁-carbazoles decrease, and those of C₂-carbazoles increase significantly, which may indicate that C₂-carbazoles are more resistant to biodegradation than lower homologous species. As to C₂-carbazole isomers, the relative concentrations of the pyrrolic N-H-shielded, pyrrolic N−H partially shielded and pyrrolic N-H-exposed isomers do not show any obvious variation in the non-heavily biodegraded oil, but there is an abrupt change through the mid-biodegraded stage to the heavily biodegraded stage. This project was financially supported by the Youth Knowledge-Innovation Foundation of CNPC (No. 00Z1304).     

Evaluation of the diagnostic ratios for the identification of spilled oils after biodegradation

Biodegradation, one of the most important weathering processes, alters the composition of spilled oil, making it difficult to identify the source of the release and to monitor its fate in the environment. A laboratory experiment was conducted to simulate oil spill weathering process of microbial degradation to investigate compositional changes in a range of source- and weathering-dependent molecular parameters in oil residues, and the conventional diagnostic ratios for oil spill identification were also evaluated. The conventional diagnostic ratios of n-alkane displayed obvious changes after biodegradation, especially for Pr/n-C₁₇ and Ph/n-C₁₈ with relative standard deviation more than 118.84 %, which suggests they are invalid for oil source identification of the middle-serious spill. Many polycyclic aromatic hydrocarbons (PAHs) are more resistant to biodegradation process than their saturated hydrocarbon counterparts, thus making PAHs to be one of the most valuable fingerprinting classes of hydrocarbons for oil identification. Biomarker ratios of hopanes and steranes were also useful for source identification even after moderate biodegradation, and the diagnostic ratios from them could be used in tracking origin and sources of hydrocarbon pollution. Finally, the carbon isotopic type curve may provide another diagnostic means for correlation and differentiation of spilled oils, and be particularly valuable for lighter refined products or severely biodegraded oils, the source of which may be difficult to identify by routine biomarker techniques.     

邻硝基对甲基苯酚和邻氨基对甲基苯酚对1,2,4-TCB厌氧生物降解影响研究

南京某化工厂生产邻硝基对甲基苯酚和邻氨基对甲基苯酚,场地调查和室内微宇宙实验研究发现该场地发生了1,2, 4-三氯苯(1,2,4-TCB)污染,且场地内天然发生着1,2,4-TCB的厌氧还原脱氯反应。为探究该场地地下水中邻硝基对甲基苯酚和邻氨基对甲基苯酚对1,2,4-TCB厌氧脱氯过程的影响,开展了3组微宇宙实验:(1)底物仅为1,2,4-TCB, (2)底物为1,2,4-TCB和邻氨基对甲基苯酚,(3)底物为1,2,4-TCB和邻硝基对甲基苯酚。对比了3种不同底物条件下1,2,4-TCB的降解过程、降解速率及微生物群落结构。实验结果表明:(1) 邻硝基对甲基苯酚在厌氧条件下还原生成邻氨基对甲基苯酚, 同时抑制1,2,4-三氯苯厌氧生物降解;最终1,2,4-TCB降解产物只有1,4-DCB。(2)邻氨基对甲基苯酚不会阻碍1,2,4-TCB的降解,但会降低降解速率。(3) Azospira和Pseudomonas与邻硝基对甲基苯酚还原反应有关,尤其是Pseudomonas极有可能是场地邻硝基对甲基苯酚还原的功能菌。     

Two rapid assays for screening of patulin biodegradation

The mycotoxin patulin is produced by the blue mould pathogen Penicillium expansum in rotting apples during postharvest storage. Patulin is toxic to a wide range of organisms, including humans, animals, fungi and bacteria. Wash water from apple packing and processing houses often harbours patulin and fungal spores, which can contaminate the environment. Ubiquitous epiphytic yeasts, such as Rhodosporidium kratochvilovae strain LS11 which is a biocontrol agent of P. expansum in apples, have the capacity to resist the toxicity of patulin and to biodegrade it. Two non-toxic products are formed. One is desoxypatulinic acid. The aim of the work was to develop rapid, high-throughput bioassays for monitoring patulin degradation in multiple samples. Escherichia coli was highly sensitive to patulin, but insensitive to desoxypatulinic acid. This was utilized to develop a detection test for patulin, replacing time-consuming thin layer chromatography or high-performance liquid chromatography. Two assays for patulin degradation were developed, one in liquid medium and the other in semi-solid medium. Both assays allow the contemporary screening of a large number of samples. The liquid medium assay utilizes 96-well microtiter plates and was optimized for using a minimum of patulin. The semi-solid medium assay has the added advantage of slowing down the biodegradation, which allows the study and isolation of transient degradation products. The two assays are complementary and have several areas of utilization, from screening a bank of microorganisms for biodegradation ability to the study of biodegradation pathways.     

The influence of biodegradation on resins and asphaltenes in the Liaohe Basin

Asphaltenes are traditionally considered to be recalcitrant to microbial alteration. Resins and asphaltenes of seven biodegraded oils extracted from reservoir cores of two columns (Es3 and Es1) of the Lengdong oilfield in the Liaohe Basin, NE China, were studied to test this hypothesis. Elemental (C, H, N, O, S) and isotopic compositions (δ¹³C and δ¹⁵N) were measured, FT-IR was used to study the oxygenated functionalities of both resins and asphaltenes, and Py–GC–MS was used to elucidate how alkyl side chains of asphaltenes were altered during biodegradation. We conclude that the products of biodegradation, such as carboxylic acids, phenol and alcohols, may not only contribute to the resin fraction of crude oils, but also are linked with functionalities of resins and asphaltenes. The amount of asphaltenes increases because some resin molecules are enlarged and their polarity increased such that they can be precipitated by hexane as newly generated asphaltenes. Thus, the hydrocarbons that are progressively consumed during biodegradation can pull the δ¹³C of asphaltene fraction closer to the δ¹³C of the altered resins and hydrocarbons that were consumed.     

煤层产甲烷菌对胍胶的生物降解实验

胍胶压裂液的低温破胶问题事关煤层的增透效果,为探讨产甲烷菌群对胍胶生物降解的影响,以内蒙古某矿褐煤样品为研究对象,以富集的新鲜矿井水为菌源,配比不同浓度的胍胶与煤的混合液,开展生物甲烷代谢实验。以产气总量、CH₄含量、胍胶分子官能团和C、H元素含量变化以及液相产物等指标表征煤层产甲烷菌群对胍胶的生物降解特性。实验结果表明：加入胍胶能提高煤制生物气产量,代谢后体系溶液黏度明显降低;降解后胍胶大分子中C、H元素的含量明显下降,胍胶的表面粗糙度增加;胍胶降解后主要特征峰表现为-OH基团及C-O基团的含量减少,甲基官能团振动吸收峰消失;生物降解溶液中葡萄糖醛酸含量增加。实验验证了煤层产甲烷菌对胍胶的降解作用及效果,为胍胶压裂液的低温破胶技术提供了有力依据。     

Evaluation of the use of legumes for biodegradation of petroleum hydrocarbons in soil

The aim of this research was to evaluate the potential of six legumes: Medicago sativa L., Glycine max, Arachis hypogea, Lablab purpureus, Pheseolus vulgaris and Cajanus cajan to restore within a short period of time soil contaminated with 3% crude oil. The legumes in five replications were grown in crude oil-contaminated and crude oil-uncontaminated soil in a completely randomized design. Plants were assessed for seedling emergence, plant height and leaf number. GC–MS was used to analyze the residual crude oil from the rhizosphere of the legumes. Plant growth parameters were reduced significantly (P < 0.05) for legumes in contaminated soil compared to their controls. In the 4th week after planting (WAP), shoot height increased across the species up to the 8th WAP. However, in the 12 WAP no significant increase in the shoot of all species was observed. Two WAP legumes planted in contaminated soil had significantly (P < 0.05) higher leaf number than these planted in uncontaminated soil with the exception of M. sativa. In the 4th WAP, only A. hypogea and P. vulgaris had increased leaf number, while in the 6th WAP, only L. purpureus had increased leaf number and survived up to the 12th WAP while most of the legumes species died. Chromatographic profiles indicated 100% degradation of the oil fractions in C. cajan and L. purpureus after 90 days. For other legumes however, greater losses of crude oil fractions C₁–C₁₀ and C₁₀–C₂₀ were indicated in rhizosphere soil of P. vulgaris and G. max, respectively. The most effective removal (93.66%) of C₂₁–C₃₀ components was observed in G. max-planted soil even though vegetation was not established. The legumes especially C. cajan, L. purpureus and A. hypogea are promising candidates for phytoremediation of petroleum hydrocarbon-impacted soil.     

荧光示踪剂的干扰实验研究

通过在室内无光照条件下开展干扰实验,探究了荧光素钠、罗丹明和荧光增白剂三种示踪剂之间的干扰规律,并应用于野外地下水二元示踪试验实例,说明了校正荧光示踪仪检测浓度(ΔC)的方法。结果表明:（1）在实验室条件下,荧光素钠的抗干扰性最强,罗丹明次之,荧光增白剂易受到干扰产生检测浓度增大的假象,这种假象服从线性变化规律;（2）当使用罗丹明作为示踪剂时,ΔC钠=0.052C罗、ΔC白=0.012C罗;当使用荧光素钠时,ΔC罗=0.507C钠、ΔC白=0.323C钠。在野外开展二元示踪试验时,建议尽量选用相互之间干扰较小的罗丹明和荧光增白剂进行组合投放,或利用本实验得到的不同示踪剂之间的干扰规律对荧光示踪仪的检测浓度进行校正。      

The role of biodegradation and photo-oxidation in the transformation of terrigenous organic matter

Microbial and photochemical decomposition are two major processes regulating organic matter (OM) transformation in the global carbon cycle. However, photo-oxidation is not as well understood as biodegradation in terms of its impact on OM alteration in terrigenous environments. We examined microbial and photochemical transformation of OM and lignin derived phenols in two plant litters (corn leaves and pine needles). Plant litter was incubated in the laboratory over 3 months and compositional changes to OM were measured using nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry. We also examined the susceptibility of soil organic matter (SOM) to ultraviolet (UV) radiation. Solid-state ¹³C NMR spectra showed that O-alkyl type structures (mainly from carbohydrates) decreased during biodegradation and the loss of small carbohydrates and aliphatic molecules was observed by solution-state ¹H NMR spectra of water extractable OM from biodegraded litters. Photochemical products were detected in the aliphatic regions of NaOH extracts from both litter samples by solution-state ¹H NMR. Photo-oxidation also increased the solubility of SOM, which was attributed to the enhanced oxidation of lignin derived phenols and photochemical degradation of macromolecular SOM species (as observed by diffusion edited ¹H NMR). Overall, our data collectively suggests that while biodegradation predominates in litter decomposition, photo-oxidation alters litter OM chemistry and plays a role in destabilizing SOM in soils exposed to UV radiation.     

Impact of anaerobic biodegradation on alkylphenanthrenes in crude oil

The biodegradation of crude oil by microorganisms from well Luo-801, China, was examined in cultures grown under conditions that promoted either methanogenesis or sulfate reduction, at 35 °C and 55 °C. Headspace gas and oil compositions were characterized at 180 d and 540 d. Alkylphenanthrenes are relatively recalcitrant to bacterial attack and the biodegradation of these compounds appeared to be insignificant after 180 d under both conditions, but is evident after 540 d. The depletion of alkylphenanthrenes was monitored through evaluation of the ratio of alkylphenanthrenes to the most bioresistant, analyzed component (C₂₈ 20R triaromatic steroid hydrocarbon) and isomer susceptibility also was evaluated by relative abundance comparison within the compound class. The influence of growth temperature varied. Only slight differences in alkylphenanthrene concentrations were observed after 180 d whereas the greater degrees of biodegradation were observed at 35 °C in the methanogenic culture and at 55 °C in the sulfate reducing culture. Overall, higher biodegradation rates occur under sulfate reducing condition, which is consistent with the conclusion that methanogens are generally less able to compete for substrates than sulfate reducers. The biodegradation susceptibility of alkylphenanthrenes decreases with increasing degree of alkylation, i.e., phenanthrene (P) and methylphenanthrenes (MPs) were more easily biodegradable than C₂-alkylphenanthrenes (C₂-Ps) and C₃-alkylphenanthrenes (C₃-Ps). Biodegradation selectivity for specific homologues is not striking for the limited time duration of the experiments. However, 3-MP seems slightly more vulnerable than other methylphenanthrene isomers and 1,7-DMP has slightly higher ability to resist biodegradation than the other C₂-P isomers. The commonly used thermal maturity parameters derived from methylphenanthrene isomer ratios are altered insignificantly by biodegradation and remain valid for geochemical assessment. This information should be useful for assessing the limits of in situ crude oil biodegradation.     

Effects of Biodegradation on Crude Oils from Karamay Oilfield

Studies of biological marker compou nds in five oil samples from a profile wherenormal crude oil,low condensate oil and heavy oil are produced in the Karamay Oilfield have been carried out with great empha-sis on the biodegradation-resisting capability of 13,17 secosteranes,8,14 secohopanes,gammacerane and carotenes.Based on these studies,a sequence of biodegradation-resisting intensities has been established for saturated hydrocarbon biomarkers in crude oils from the Karamay Oilfield.     

Hydrogen peroxide interference in chemical oxygen demand during ozone based advanced oxidation of anaerobically digested livestock wastewater

It is known that hydrogen peroxide interferes with chemical oxygen demand analysis by consuming oxidation agents such as potassium dichromate, thus leading to overestimation of the chemical oxygen demand measurements. The objective of the study was to investigate the effects of hydrogen peroxide interference and to determine true chemical oxygen demand values on interpreting treatment performance during ozone-based advanced oxidation of livestock wastewater in which hydrogen peroxide concentration and chemical oxygen demand values are dynamically changing. According to the chemical oxygen demand monitoring data, chemical oxygen demand values were always higher than the initial chemical oxygen demand load when hydrogen peroxide was involved and the treatment performance with ozone alone or ozone/ultraviolet was better than with coupled hydrogen peroxide. The extent of overestimation was proportional to the remaining hydrogen peroxide concentration and the average overestimation ratio in livestock wastewater was in the range of 0.50～0.58 mg per 1 mg of hydrogen peroxide, depending upon the quality of the wastewater treated. True chemical oxygen demand values were estimated by correlating the extent of overestimation with the remaining hydrogen peroxide concentration during treatment. The extent of overestimation decreased to zero gradually as the amount of hydrogen peroxide also approached zero as oxidation proceeded. The corrected chemical oxygen demand values indicated underlying tendency of oxidation, which could not be seen in the original chemical oxygen demand monitoring data. Application of ozone/hydrogen peroxide was more efficient for reducing chemical oxygen demand than ozone alone, as was ozone/hydrogen peroxide/ultraviolet compared to ozone/ultraviolet. When coupled with ozone, ultraviolet irradiation was more efficient than hydrogen peroxide for decreasing chemical oxygen demand during treatment of livestock wastewater.