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Production of biodiesel fuel by transesterification of rapeseed oil.

Fatty acid methyl esters (FAMEs) show large potential applications as diesel substitutes, also known as biodiesel fuel. Biodiesel fuel as renewable energy is an alternative that can reduce energy dependence on petroleum as well as air pollution. Several processes for the production of biodiesel fuel have been developed. Transesterification processes under alkali catalysis with short-chain alcohols give high yields of methyl esters in short reaction times. We investigated transesterification of rapeseed oil to produce the FAMEs. Experimental reaction conditions were molar ratio of oil to alcohol, concentration of catalyst, type of catalyst, reaction time, and temperature. The conversion ratio of rapeseed oil was enhanced by the alcohol:oil mixing ratio and the reaction temperature.

Alcohols↗

Microdiesel: Escherichia coli engineered for fuel production.

Biodiesel is an alternative energy source and a substitute for petroleum-based diesel fuel. It is produced from renewable biomass by transesterification of triacylglycerols from plant oils, yielding monoalkyl esters of long-chain fatty acids with short-chain alcohols such as fatty acid methyl esters and fatty acid ethyl esters (FAEEs). Despite numerous environmental benefits, a broader use of biodiesel is hampered by the extensive acreage required for sufficient production of oilseed crops. Therefore, processes are urgently needed to enable biodiesel production from more readily available bulk plant materials like sugars or cellulose. Toward this goal, the authors established biosynthesis of biodiesel-adequate FAEEs, referred to as Microdiesel, in metabolically engineered Escherichia coli. This was achieved by heterologous expression in E. coli of the Zymomonas mobilis pyruvate decarboxylase and alcohol dehydrogenase and the unspecific acyltransferase from Acinetobacter baylyi strain ADP1. By this approach, ethanol formation was combined with subsequent esterification of the ethanol with the acyl moieties of coenzyme A thioesters of fatty acids if the cells were cultivated under aerobic conditions in the presence of glucose and oleic acid. Ethyl oleate was the major constituent of these FAEEs, with minor amounts of ethyl palmitate and ethyl palmitoleate. FAEE concentrations of 1.28 g l(-1) and a FAEE content of the cells of 26 % of the cellular dry mass were achieved by fed-batch fermentation using renewable carbon sources. This novel approach might pave the way for industrial production of biodiesel equivalents from renewable resources by employing engineered micro-organisms, enabling a broader use of biodiesel-like fuels in the future.

Alcohols↗

Anaerobic benzene degradation.

Although many studies have indicated that benzene persists under anaerobic conditions in petroleum-contaminated environments, it has recently been documented that benzene can be anaerobically oxidized with most commonly considered electron acceptors for anaerobic respiration. These include: Fe(III), sulfate, nitrate, and possibly humic substances. Benzene can also be converted to methane and carbon dioxide under methanogenic conditions. There is evidence that benzene can be degraded under in situ conditions in petroleum-contaminated aquifers in which either Fe(III) reduction or methane production is the predominant terminal electron-accepting process. Furthermore, evidence from laboratory studies suggests that benzene may be anaerobically degraded in petroleum-contaminated marine sediments under sulfate-reducing conditions. Laboratory studies have suggested that within the Fe(III) reduction zone of petroleum-contaminated aquifers, benzene degradation can be stimulated with the addition of synthetic chelators which make Fe(III) more available for microbial reduction. The addition of humic substances and other compounds that contain quinone moieties can also stimulate anaerobic benzene degradation in laboratory incubations of Fe(III)-reducing aquifer sediments by providing an electron shuttle between Fe(III)-reducing microorganisms and insoluble Fe(III) oxides. Anaerobic benzene degradation in aquifer sediments can be stimulated with the addition of sulfate, but in some instances an inoculum of benzene-oxidizing, sulfate-reducing microorganisms must also be added. In a field trial, sulfate addition to the methanogenic zone of a petroleum-contaminated aquifer stimulated the growth and activity of sulfate-reducing microorganisms and enhanced benzene removal. Molecular phylogenetic studies have provided indications of what microorganisms might be involved in anaerobic benzene degradation in aquifers. The major factor limiting further understanding of anaerobic benzene degradation is the lack of a pure culture of an organism capable of anaerobic benzene degradation.

Bacteria, Anaerobic↗

Effect of nitrates on biotransformation of phosphogypsum and phenol uptake in cultures of autochthonous sludge microflora from petroleum refining wastewaters.

The effect of nitrates on the biotransformation of phosphogypsum at 30 degrees C in stationary cultures of anaerobic, heterogeneous microflora growing in medium with phenol (250-1,000 mg/L) as sole carbon source was studied. The microorganisms used in this study were isolated from sludge in biological petroleum-refining wastewater treatment plant. Phosphogypsum (a waste product in the chemical industry that contains approximately 95% CaSO4) was added in amount of 5 g/L, the source of nitrates was KNO3 in concentration equivalent to that of phenol (250-1,000 mg N-NO3/L). The presence of nitrates in heterogeneous cultures has an inhibitory effect on the process of phosphogypsum biotransformation and stimulates the uptake of phenol. We have found that in cultures in medium containing phenol, phosphogypsum and nitrates at least three physiological groups of microorganisms were present. These were phenol-biodegrading microorganisms not requiring an external electron acceptor, sulfate-reducing bacteria biodegrading phenol or intermediate products of its breakdown and denitrifying bacteria not utilising phenol as a carbon source. On solid medium these bacteria together formed heterogeneous single colonies. In spite of repeated attempts we were unable to isolate pure strains and the only result of these measures was loss of denitrification ability in medium with phenol.

Bacteria, Anaerobic↗

Enzymatic conversion of all-trans-beta-carotene to retinal.

Enzymatic conversion of all-trans-beta-carotene to retinal by a partially purified enzyme from rabbit, rat, and human neonatal intestinal mucosa has been demonstrated. The enzymatic product was characterized based on the following evidence. First, the product gave rise to its O-ethyl oxime by treatment with O-ethylhydroxylamine with an absorption maximum at 363 nm in ethanol characteristic of authentic retinal (O-ethyl) oxime. High-performance liquid chromatography of this derivative yielded a sharp peak with a retention time of 7.99 min, corresponding to the authentic compound. The enzyme blank and boiled enzyme blank failed to show any significant HPLC peaks corresponding to retinal (O-ethyl) oxime or retinal or retinol. Second, the mass spectrum of the O-ethyl oxime of the enzymatic product was identical to that of authentic retinal (O-ethyl) oxime (m/z 327, 45%; m+ and m/z 282, 100%, methoxy). Third, the 14C radioactivity persisted to constant specific activity even after repeated crystallization of the retinal (O-ethyl) oxime isolated from the enzyme reaction with purified beta-[14C]carotene. Fourth, the enzymatic product exhibited an absorption maximum at 370 nm in light petroleum characteristic of authentic retinal. Furthermore, it was reduced by horse liver alcohol dehydrogenase to retinol with an absorption maximum at 326 nm in light petroleum. This retinol was enzymatically esterified to retinyl palmitate by rat pancreatic esterase with a retention time of 10 min on HPLC, corresponding to authentic retinyl palmitate. Thus, the enzymatic product of beta-carotene cleavage by the partially purified intestinal enzyme has been unequivocally confirmed to be retinal. Similarly, enzymatic conversion of all-trans-beta-carotene to retinal by an intestinal mucosal enzyme from autopsy samples of human neonates has also been demonstrated. Based on the observed activities among intestinal samples from 12 premature infants, the BCC enzyme activity ranged from 3.3 to 1210 pmol/mg mucosal protein/hr. However, the observed activities in the human autopsy samples may be markedly underestimated, presumably because of marked loss of enzyme activity from the time of death to the time of assay. Therefore, the true activity of the enzyme can be assessed only after the extent of the loss of its activity on storage of the human samples can be accurately measured. Nonetheless, the demonstration of BCC enzyme activity in human neonates shows that beta-carotene may be an important source of vitamin A nutrition during gestation.

Animals↗

Enzymatic conversion of all-trans-beta-carotene to retinal by a cytosolic enzyme from rabbit and rat intestinal mucosa.

Enzymatic conversion of all-trans-beta-carotene to retinal by a partially purified enzyme from rabbit and rat intestinal mucosa was demonstrated. The enzymatic product was characterized based on the following evidence: (i) The product gave rise to its O-ethyloxime by treatment with O-ethylhydroxylamine with an absorption maximum at 363 nm in ethanol characteristic of authentic retinal O-ethyloxime. High-pressure liquid chromatography (HPLC) of this derivative yielded a sharp peak with a retention time of 7.99 min corresponding to the authentic compound. The enzyme blank and boiled enzyme blank failed to show any significant HPLC peaks corresponding to retinal O-ethyloxime, retinal, or retinol. (ii) The mass spectrum of the O-ethyloxime of the enzymatic product was identical to that of authentic retinal O-ethyloxime (m/z 327: 45%, M+. and m/z 282: 100%, M--ethoxy). (iii) The specific activity of the enzymatically formed [14C]retinal O-ethyloxime remained constant even after repeated crystallization. (iv) The enzymatic product exhibited an absorption maximum at 370 nm in light petroleum characteristic of authentic retinal. Furthermore, it was reduced by horse liver alcohol dehydrogenase to retinol with an absorption maximum at 326 nm in light petroleum. This retinol was enzymatically esterified to retinyl palmitate by rat pancreatic esterase with a retention time of 10 min on HPLC corresponding to authentic retinyl palmitate. Thus, the enzymatic product of beta-carotene cleavage by the partially purified intestinal enzyme was unequivocally confirmed to be retinal.

Animals↗

Fabric protectors. Part II - Propane, 1,1,1-trichloroethane, and petroleum distillates levels in air after application of fabric protectors.

Propane, 1,1,1-trichloroethane, and petroleum distillates levels in air which were generated during the use of aerosol type fabric protectors were monitored by means of the NIOSH charcoal tube, a glass bulb grab sampling, and the GASBADGE passive device techniques. Although 1982 ACGIH TLV-STEL were readily exceeded in an unventilated test room, when fabric was sprayed with 450 g of fabric protector in an unconfined area within a home the generated vapors quickly dispersed and STEL and 8-hour TWA-TLV were not exceeded.

Air Pollutants↗

Contaminant risks from biosolids land application: contemporary organic contaminant levels in digested sewage sludge from five treatment plants in Greater Vancouver, British Columbia.

This study examines the potential for environmental risks due to organic contaminants at sewage sludge application sites, and documents metals and various potential organic contaminants (volatile organics, chlorinated pesticides, PCBs, dioxins/furans, extractable petroleum hydrocarbons, PAHs, phenols, and others) in current production biosolids from five wastewater treatment plants (WWTPs) within the Greater Vancouver Regional District (GVRD). There has been greater focus in Europe, North America and elsewhere on metals accumulation in biosolids-amended soil than on organic substances, with the exception of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans. Another objective, therefore, was to evaluate the extent to which management of biosolids re-use based on metal/metalloid levels coincidentally minimizes environmental risks from organic contaminants. Historical-use contaminants such as chlorophenols, PCBs, and chlorinated pesticides were not detected at environmentally relevant concentrations in any of the 36 fresh biosolids samples, and appear to have virtually eliminated from sanitary collection system inputs. The few organic contaminants found in freshly produced biosolids samples that exhibited high concentrations relative to British Columbia and Canadian soil quality benchmarks included p-cresol, phenol, phenanthrene, pyrene, naphthalene, and heavy extractable petroleum hydrocarbons (HEPHs--nCl9-C34 effective carbon chain length). It was concluded that, with the exception of these petroleum hydrocarbon constituents or their microbial metabolites, the mixing of biosolids with uncontaminated soils during land application and based on the known metal concentrations in biosolids from the Greater Vancouver WWTPs investigated provides adequate protection against the environmental risks associated with organic substances such as dioxins and furans, phthalate esters, or volatile organics. Unlike many other organic contaminants, the concentrations of petroleum hydrocarbon derived substances in biosolids has not decreased within the last decade or more in the WWTPs studied, and--unlike persistent chlorinated compounds--the associated PAHs and other hydrocarbon constituents merit careful consideration, especially in the context of repeated land-application of biosolid.

Canada↗

Low salt petroleum produced water reuse: a farming alternative outside the food chain.

Oil and gas production gives rise to water production depending on the state of maturation of the Field. This means large volumes of water available. Today, this water is partly re-injected into the reservoir. Totalfinaelf's sustainable contribution to preserve water resources is to propose an alternative utilization: the reuse of produced waters from petroleum activities, outside the food chain. The aim of the first part of this study is to demonstrate the feasibility of utilizing low-salt water (<20 g/l) cleaned of hydrocarbons, for agricultural or forest irrigation. Rudimentary technologies such as artificial wetlands were tested to remove hydrocarbon substances and preliminary tests were performed with cotton (Gossypium hirsutum) and hemp (Cannabis sativa). Among the species tested in real conditions (greenhouse), hemp was affected by saline water whereas the results obtained for cotton were representative of the average worldwide production. These results validate the reuse of low-salt produced water in climatic conditions with expected temperatures of up to 37 degrees C in summer and 25 degrees C in winter. Following these results, field pilots are planned. Further research is planned to focus, taking into account local needs and environmental and production constraints.

Agriculture↗

Respirometry for assessing the biodegradation of petroleum hydrocarbons.

The respiration method using the Micro-Oxymax respirometer was applied to evaluate the bioremediation potential of hydrocarbon-contaminated soils in two biopiles at the oil refinery in Czechowice-Dziedzice, Poland. In biopiles 1 and 2, two different technologies, i.e., enhanced (engineered) bioremediation and monitored natural attenuation (MNA) were used, respectively. In biopiles 1 and 2, the bioremediation process lasted 6 years and 8 months, respectively. The biodegradation of petroleum hydrocarbons was evaluated on the basis of CO2 production and O2 uptake. The CO2 production and O2 consumption rates during hydrocarbon biodegradation were calculated from the slopes of cumulative curve linear regressions. The results confirmed the hydrocarbon biodegradation process in both biopiles. However, in biopile 2 the process was more effective compared to biopile 1. In biopile 2, the O2 consumption and CO2 production means were 3.37 and 2.4 milliliters per kilogram of soil (dry weight) per minute, respectively. Whereas, in biopile 1, the O2 consumption and CO2 production means were 1.52 and 1.07 milliliters per kilogram of soil (dry weight) per minute, respectively. The mean biodegradation rate for biopile 2 was two times higher--67 mg hydrocarbons kg d.w.(-1)day(-1) compared with biopile 1, where the mean was 30 mg hydrocarbons kg d.w.(-1)day(-l). The results were correlated with petroleum hydrocarbon concentrations and microbial activity measured by dehydrogenase assay.

Biodegradation, Environmental↗

Acute toxicity of vanadium to the threespine stickleback, Gasterosteus aculeatus.

Vanadium is widely distributed, occurring in many types of minerals, coal, and petroleum. Anthropogenic sources of vanadium originate from the production, processing, and wastes of these materials. The aquatic toxicity of vanadium to fish species is not well characterized. This study focused on the three-spined stickleback, Gasterosteus aculeatus, a small and widely distributed euryhaline species of fish. The three-spined stickleback is used as an effluent-monitoring species in both Canada and the United States. Five 96-h static renewal acute toxicity tests were performed in moderately hard water with adult fish. The geometric mean and range of the five 96-h LC(50)s based on measured concentrations of total vanadium in the test solution were 3.17 and 2.35-4.07 mg V/L, respectively. A conservative estimation of a safe concentration of vanadium that would not affect survival of adult three-spined sticklebacks over a 96-h exposure period in moderately hard water is approximately 0.30 mg V/L. A comparison with other fish species previously tested suggests that the three-spined stickleback is intermediate in sensitivity to vanadium. Information reported from this study may be useful in effluent toxicity identification evaluations and ecological risk assessments related to vanadium.

Animals↗

Prevalence of abnormal neurobehavioral scores in populations exposed to different industrial chemicals.

The aim of the study is to establish the prevalence of neurobehavioral scores of occupationally exposed subjects below the 10th percentile rank of normalized curves obtained on a referent population. The Milan Automated Neurobehavioral System (MANS) was administered to 400 drivers from public and private firms; their data were distributed on the basis of age and years of school attendance and were normalized by determining percentile rank equivalence. The exposed population is made up of 20 lead- and zinc-exposed subjects, 18 welders exposed to aluminum for less than 1 year, 150 exposed to different metals in the ferromanganese production, 73 lithographic operators exposed to gasoline and petroleum, 197 exposed to solvents mixtures in the paint manufacture, and 23 dropouts of the same firm. The percentages of scores below 10th percentile rank were calculated in the different exposure groups and in the different age-school attendance ranges. The prevalence of results below the 10th percentile rank was found to be related to the intensity of the exposures and to the low levels of school attendance. In the 20-29 and the 30-39 age ranges, there was a prevalence of POMS scale scores below 10th percentile rank, in the 50-59 age range, the percentages were high for the Digit Symbol, the mean value of Simple Reaction Time, Serial Digit Learning, and Benton Visual Recognition.

Adult↗

The genotoxicity of industrial wastes and effluents.

A review of the literature published on the genotoxicity of industrial wastes and effluents using short-term genetic bioassays is presented in this document. The importance of this task arises from the ubiquity of genotoxic compounds in the environment and the need to identify the sources of contamination so that efforts aimed at control and minimization can be implemented. Of even greater significance is the immediate concern for the welfare of human health and the environment. Subheadings of this document include a description of the genetic bioassays that have been used to test industrial wastes, a compendium of methods commonly used to prepare crude waste samples for bioassay, and a review of the genetic toxicity of wastes and effluents. Wastes and effluents have been grouped according to industrial source. Major categories include chemical and allied products, pulp and paper manufacturing, defense and munitions, petroleum refining, primary metal industries, and miscellaneous industrial manufacturers. Within each industrial category, a synopsis of individual genetic toxicity studies is presented, followed by an interpretation of results on a comprehensive, industry-wide basis. In this evaluation, a discussion of the types and extent of genotoxic damage caused by a particular set of wastes is presented, and potential sources of genotoxic activity are identified. Concluding the document is a commentary, which discloses potential shortcomings in the way in which current legislation protects human heath and the environment from the release of genotoxic substances via industrial wastes and effluents. It also provides an assessment of the genotoxic burden that industrial wastes place on the environment.

Industrial Waste↗

Common household poisonings.

Readily available household products represent a source of potential toxicity when ingested accidentally by children. Despite the large number of patients seen by physicians and a familiarity with many of these substances, patient management often remains a difficult problem. The current literature is reviewed with respect to appropriate management of ingestions of household cleaners, caustics, hydrocarbons, insecticides, rodenticides, plants, and other products often taken by children.

Child↗

[Determination of vitamin D2 included with beta-cyclodextrin complex in "longmu zhuanggu chongji" by HPLC].

In order to enhance the stability of vitamin D2(VD2), the inclusion complex of VD2 with beta-cyclodextrin(beta-CD) was added in "Longmu Zhuanggu Chongji", a medicine of children for the prevention and cure of rickets. The VD2 in beta-CD-VD2 inclusion complex was satisfactorily determined by both reversed-phase high performance liquid chromatography(HPLC) and ultraviolet spectrometry(UV). The experimental results from UV were in well consistent with those from HPLC. The accuracy and recovery of HPLC measurement were analyzed, and their relative standard deviation(RSD) were given to be 0.63% and 1.54%, indicating the method is quite correct and reliable, and the linear regression equation is reasonable. The "Longmu Zhuanggu Chongji" containing beta-CD-VD2 inclusion complexes was saponified by a mixture of KOH, vitamin C (VC) and alcohol-water(1:2, V/V) at 70 degrees C, then extracted with petroleum ether. An accurate content of the VD2 product was obtained by HPLC. A simple and rapid HPLC method, which can detect VD2 in the inclusion complex and in "Longmu Zhuanggu Chongji", has been established in this work.

Chromatography, High Pressure Liquid↗

[Low-temperature microbial degradation of crude oil products differing in the extent of condensation].

Out of the 30 strains capable of oil degradation at 4-6 degrees C, four were selected by the ability to degrade 40% of the oil substrate present in the growth medium: Rhodococcus spp. DS-07 and DS-21 and Pseudomonas spp. DS-09 and DS-22. We studied the activity of these strains as degraders of oil products of various condensation degrees (crude oil, masut, petroleum oils, benzene resins and ethanol-benzene resins) at 4-6 degrees C. The maximum degrees of degradation of masut and ethanol-benzene resins were observed in Pseudomonas spp. DS-22 (17.2% and 5.2%, respectively). The maximum degradation of petroleum oils and benzene resins was observed in Rhodococcus spp. DS-07 (40% and 16.6%, respectively). The strains provide a basis for developing biodegrader preparations applicable to bioremediation of oil-polluted sites under the conditions of cold climate.

Benzene↗

[Ingestion of undesirable substances by children].

A retrospective study extending over five years was made of the consequences of ingestion of undesirable substances by children. Despite the publicity on this subject it remains a serious medical problem. An annual average of 90 children were seen in the emergency department. Complications are caused mostly by oil products but also by drugs and household products. On average 55% of the children were hospitalized. With regard to the indication for endoscopical examination of the oesophagus and stomach after ingestion of caustic substances, insufficient arguments for performance of this examination in all children were found in the literature. We advocate a prospective study of this question. With regard to the indication for hospitalization of children after ingestion of tobacco, we are of the opinion that many children may be kept under observation in their homes after being watched for one hour.

Burns, Chemical↗