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Comparative dermal carcinogenesis of shale and petroleum-derived distillates.

Ten test materials derived from petroleum or hydrotreated shale oils were applied 3 times/week for up to 105 weeks to the shaved skin of 25 male and 25 female C3H/HeN mice per group. Mineral oil and benzo(a) pyrene (0.15%) were control materials. Clinical observations were recorded during the study. At death, histopathologic examination was conducted on skin, internal organs and any gross lesions. Exposures to some materials were ended midway in the study due to severe irritation. Chronic toxicity of all materials was limited to inflammatory and degenerative skin changes. Significant increases over control incidence of skin tumors (squamous cell carcinoma and fibrosarcoma) occurred with both petroleum and shale-derived naphtha (21%, 50%), Jet A (26%, 28%), JP-4 (26%, 50%), and crude oils (84%, 54%). Severely hydrotreated shale oil and petroleum and shale-derived diesel distillates were not considered tumorigenic. Results indicate that toxicity of comparable petroleum and shale-derived fractions was qualitatively similar and confirm earlier findings that hydrotreating reduces or eliminates carcinogenicity of raw shale oil.

Animals↗

Statistical association between cancer incidence and major-cause mortality, and estimated residential exposure to air emissions from petroleum and chemical plants.

An ecologic study design was used to investigate the relationship between exposure to air emissions produced by the petroleum and chemical industries, and average annual cancer incidence and major cause mortality rates among whites in Contra Costa County, California. Estimates for the exposure to major industrial sources of sulfur dioxide, hydrocarbons and oxides of nitrogen were used to subdivide the county by level of exposure to petroleum refinery and chemical plant emissions. Cancer incidence and major cause mortality rates were then calculated for whites in each of the exposure areas. In both males and females, residential exposure to petroleum and chemical air emissions was associated with an increased incidence of cancer of the buccal cavity and pharynx. In males, age-adjusted incidence rates for cancers of the stomach, lung, prostate and kidney and urinary organs were also associated with petroleum and chemical plant air emission exposures. In both sexes, we found a strong positive association between degree of residential exposure and death rates from cardiovascular disease and cancer, and a less strong positive association between exposure and death rates from cerebrovascular disease. There was also a positive association in men for deaths from cirrhosis of the liver. Although these observed associations occurred across areas of similar socioeconomic and broad occupational class, confounding variables and the "ecologic fallacy" must be considered as possible explanations. In particular, the stronger findings in men suggest an occupational explanation of the cancer incidence trends, and the effect observed in cirrhosis mortality suggests that lifestyle variables such as alcohol consumption were not adequately controlled for. While the public health implications of our findings remain unclear, the evidence presented is sufficient to warrant follow-up studies based on individual data in which possible biases can be more readily controlled.

Air Pollutants↗

Effect of petroleum-containing wastewater irrigation on bacterial diversities and enzymatic activities in a paddy soil irrigation area.

Effects of petroleum contamination on bacterial diversities and enzymatic activities in paddy soils were investigated in the Shenfu irrigation area, the largest area irrigated by oil-containing wastewater for more than 50 yr in northeastern China. Bacterial diversities were determined by conventional colony morphology typing techniques and 16S rDNA polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE). Dehydrogenase, hydrogen peroxidase, polyphenol oxidase, urease, and substrate-induced respiration (SIR) were measured to evaluate the effects of petroleum-containing wastewater irrigation on soil biochemical characteristics. Results showed that paddy soil total petroleum hydrocarbon (TPH) concentration in the irrigation area varied from 277.11 to 5213.37 mg kg(-1) dry soil. Soil TPH concentration declined along the gradient of the irrigation channel from up- to downstream. At the current pollution level, the paddy soil TPH concentration was positively correlated with the colony forming units (CFU) of aerobic heterotrophic bacteria (AHB) (r = 0.928, p < 0.001) and the genetic diversity based on DGGE profiles (r = 0.655, p < 0.05). The bacterial diversities in the soils based on colony morphotypes of AHB also increased with TPH concentration (r = 0.598), but not significant statistically (p = 0.052). Analysis of soil enzyme activities indicated a significant positive correlation between soil TPH concentration and activities of dehydrogenases (r = 0.974, p < 0.001), hydrogen peroxidases (r = 0.957, p < 0.001), polyphenol oxidases (r = 0.886, p < 0.001), and SIR (r = 0.916, p < 0.001). On the contrary, the urease activity showed a negative correlation with paddy soil TPH concentration (r = -0.814, p = 0.002), and could be used as a sensitive indicator of petroleum contamination.

Agriculture↗

Evaluation of changes in hematologic and clinical biochemical values after exposure to petroleum products in mink (Mustela vison) as a model for assessment of sea otters (Enhydra lutris).

OBJECTIVE: To determine the effects of petroleum exposure on hematologic and clinical biochemical results of mink and to identify variables that may be useful for making management decisions involving sea otters (Enhydra lutris) that have been exposed to oil in their environment. ANIMALS: 122 American mink (Mustela vison). PROCEDURES: Mink were exposed once to a slick of oil (Alaskan North Slope crude oil or bunker C fuel oil) on seawater or via low-level contamination of their daily rations. RESULTS: In the acute phase of exposure, petroleum directly affected RBC, WBC, neutrophil, and lymphocyte counts, fibrinogen, sodium, calcium, creatinine, total protein, and cholesterol concentrations, and alanine transaminase, creatine kinase, alkaline phosphatase, and gamma-glutamyltransferase activities. Aspartate transaminase, alkaline phosphatase, gamma-glutamyltransferase, and lactate dehydrogenase activities and cholesterol concentration also varied as a result of chronic low-level contamination of feed. CONCLUSIONS AND CLINICAL RELEVANCE: Our results are in agreement with reports that attribute increased alanine transaminase and alkaline phosphatase activities and decreased total protein concentration to petroleum exposure in sea otters during an oil spill. Sodium, calcium, creatinine, cholesterol, and lactate dehydrogenase may be valuable variables to assess for guidance during initial treatment of sea otters exposed to oil spills as well as for predicting which petroleum-exposed sea otters will reproduce following an oil spill. Measurement of these variables should aid wildlife professionals in making decisions regarding treatment of sea otters after oil spills.

Animals↗

Interlaboratory evaluation of an off-line supercritical fluid extraction/infrared spectrometric method for determination of petroleum hydrocarbons in solid matrixes.

A collaborative study was conducted, with 14 laboratories participating, to determine the method accuracy and precision of the proposed U.S. Environmental Protection Agency Methods 3560 and 8440. These methods involve the extraction of petroleum hydrocarbons from solid matrixes with supercritical carbon dioxide at 340 atm and 80 degrees C for 30 min (dynamic), collection of the extracted materials in tetrachloroethene (Method 3560), and analysis of the extracts by infrared (IR) spectrometry (Method 8440). The study design was based on the AOAC blind replicate design with balanced replicates. The study samples consisted of 4 solid matrixes that had petroleum hydrocarbon contents ranging from 614 to 32,600 mg/kg. Each of the 4 matrixes was extracted in triplicate, and the extracts were analyzed with 2 different IR spectrometers. In addition, each of the participating laboratories extracted a sample of unspiked clay soil, the same clay soil spiked with corn oil and reference oil at 1000 mg/kg each, and the same clay soil wetted to 30% water content and spiked with motor oil at 10,000 mg/kg (the latter 3 samples were extracted only once). Results indicated that the overall method accuracy for concentrations ranging from 614 to 32,600 mg/kg was 82.9%; the mean recoveries of petroleum hydrocarbons for each of the 4 solid matrixes ranged from 77.9 to 107% for analyses performed with the Perkin-Elmer Fourier transform IR spectrometer and from 75.9 to 101% for analyses performed with the Buck-Scientific IR spectrometer; the differences between the 2 instruments on a sample-by-sample basis were less than 17% for the total petroleum hydrocarbon determinations. The interlaboratory method precisions (RSDR) appeared to be matrix-dependent and ranged from 17.3 to 45.4% for analyses performed with the Perkin-Elmer Fourier transform IR spectrometer and from 16.7 to 47.9% for the Buck-Scientific IR spectrometer. The intralaboratory method precisions (RSDr) appeared to be less matrix-dependent and ranged from 11.5 to 17.0% for analyses performed with the Perkin-Elmer FTIR spectrometer and from 11.1 to 18.2% for the Buck-Scientific IR spectrometer. Method accuracy and precision data are also presented for the 5 laboratories that used Isco supercritical fluid extraction systems and for the 7 laboratories that used vessels with volumes of 3.5 mL or less with different supercritical fluid extraction systems.

Aluminum Silicates↗

Induction of short-term biomarkers of tumor promotion in skin of CD-1 mice by petroleum middle distillates: preliminary observations.

The induction of sustained epidermal hyperplasia in mouse skin has been shown to be a reliable predictor of tumor promoting activity for chemicals as diverse as phorbol esters, anthralins, n-dodecane and organic peroxides (DiGiovanni, 1992). The results contained herein demonstrate that API 81-07 and API 81-10, petroleum middle distillates that exhibit tumor promoting activity in mouse skin, induce epidermal hyperplasia and ODC activity. Other petroleum middle distillates (odorless light petroleum hydrocarbons, a light vacuum distillate, and a mineral seal oil) were also shown to share these activities. It should be emphasized that the relevance of these observations to human skin remains unresolved; however, the availability of these short-term biomarkers offers the opportunity to address the issue by performing comparative investigations on the effects of petroleum middle distillates on human skin xenografted to athymic (nude) mice. Such studies are being initiated.

Alkanes↗

The petroleum ether extract of Nigella sativa exerts lipid-lowering and insulin-sensitizing actions in the rat.

We studied the effect of a 4-week intragastric gavage with a petroleum ether extract of Nigella sativa seeds on blood glucose, insulin and lipids in the normal rat. Petroleum ether extract caused a 25% reduction in food intake that translated into a transient weight loss. No sign of toxicity of the plant could be seen in vivo or in vitro. Fasting plasma glucose remained stable throughout Nigella sativa treatment. At the end of the 4-week treatment, Nigella sativa-treated rats had lower fasting plasma levels of insulin and triglycerides, and higher HDL-cholesterol as compared to pair-fed controls. Response to insulin was evaluated in hepatocytes isolated from animals of all groups by Western blot analysis of phosphorylated MAPK p44/42erk and PKB. In vivo Nigella sativa treatment resulted in greater dose-dependent activation of MAPK and PKB in response to insulin. These results suggest that the petroleum ether extract of Nigella sativa has a slight anorexic effect, and that it contains the hypolipidemic activity previously obtained with the plant. More significantly, our data demonstrate that in vivo treatment with the petroleum ether extract exerts an insulin-sensitizing action by enhancing the activity of the two major intracellular signal transduction pathways of the hormone's receptor.

Alkanes↗

Direct determination of soil surface-bound polycyclic aromatic hydrocarbons in petroleum-contaminated soils by real-time aerosol mass spectrometry

Soil surface-bound polycyclic aromatic hydrocarbons (PAHs) were identified by use of Real-Time Aerosol Mass Spectrometry (RTAMS) in two NIST standard research material (SRM) soils (Montana SRM 2710 and Peruvian SRM 4355) each contaminated separately with three common petroleum hydrocarbons (diesel fuel, gasoline, and kerosene). The described contaminated soil analysis required no sample preparation. Direct laser desorption/ionization mass spectrometric analysis of individual soil particles contaminated with each of the petroleum hydrocarbons at three different contamination levels (0.8, 8, and 80 ppth (wt/wt)) yielded detectable PAH cation distributions that ranged from m/z 128 to 234, depending on the fuel contaminant. The same analysis performed on uncontaminated SRM soils revealed very little (Peruvian) to no (Montana) detectable PAH species. Size analysis showed that most of the individual soil particles analyzed were between 1 and 5 microm in diameter. Tandem mass spectrometry (MS/MS) experiments identified alkyl-substituted two- and three-ringed PAHs in all three petroleum hydrocarbon contaminated soils. However, due to similarities in fragmentation patterns, MS/MS analysis of higher MW species (m/z > 200) was unable to distinguish between the possibility of highly alkyl-substituted three-ringed PAHs and hydrogenated four-ringed PAHs. The described technique offers the direct, rapid determination and characterization of surface-bound PAHs in petroleum-contaminated soils at part-per-million levels without prior extraction, separation, or other sample preparation methods.

Journal Article↗

Effect of petroleum derivatives and solvents on colour perception.

BACKGROUND: Occupational exposure to various neurotoxic chemicals has been shown to be associated with colour vision impairment. It seems that this can occur at low exposure levels, sometimes well below the recommended occupational threshold limits. This study was undertaken to determine the effect of exposure to petroleum derivatives (polyethylene, polystyrene) and solvents (perchloroethylene) on colour perception. METHODS: Colour vision was assessed using the Ishihara plates, the D-15 test and the Farnsworth Munsell 100 Hue test. Two factories using petroleum derivatives and three dry cleaning premises were chosen at random. A total of 93 apparently healthy employees were recruited from the five workplaces. Two age-matched control groups comprising 56 people, who were support staff of the university with no exposure to petroleum, solvents or their derivatives, were also recruited. RESULTS: All subjects passed the Ishihara test, showing that none had a congenital red-green defect. Some of the exposed employees failed the D-15 and had abnormally high FM100 Hue scores. All control subjects passed all the colour vision tests. The D-15 test showed that 28 per cent (26 of 93) of exposed employees had a colour vision defect whereas the FM 100 Hue test found that 63 per cent (59 of 93) had a colour vision defect. Most defects were of the blue-yellow type (22.6 per cent) when using the D-15 test. However, with the FM 100 Hue test, most defects were of the non-polar type with no specific axis (50.5 per cent). Mean total error scores calculated from the FM100 Hue test for exposed employees were statistically significantly higher than those of the control subjects. CONCLUSION: Employees directly exposed to petroleum derivatives and solvents have a higher risk of acquiring colour vision defects compared to subjects who are not.

Adult↗

Sister chromatid exchanges in lymphocytes of petroleum retailers.

Occupational exposure to petroleum vapour was assessed in workers employed in suburban petroleum retail outlets. Urinary output of thioethers provided a non-specific estimate of exposure to chemicals metabolised via a mercapturic acid pathway. Urinary d-glucaric acid (DGA) excretion was taken as an estimate of hepatic enzyme activity. Sister chromatid exchange frequency in lymphocytes was used as an indicator of genotoxic response to exposure. Workers were classified according to their employment at self service (where customers operate petrol pumps) or at driveway attended service stations (at which an employee operates the pumps), and according to exposure to cigarette smoke on the basis of urinary cotinine excretion. Prework and postwork urine samples of workers employed at driveway attended petrol stations contained more thioether than did those of self serve workers. When classified according to smoking behaviour there were no statistically significant differences, although thioether excretion tended to be higher in smokers than in nonsmokers. Urinary DGA excretion was similar in the two exposure groups. Cigarette smokers excreted more DGA, however, than nonsmokers. Sister chromatid exchange frequencies were higher in driveway attendants than in self serve personnel. When the influence of cigarette smoking was investigated there was a significant increase of sister chromatid exchange with combined exposure to petrol and cigarette smoking, but not with either factor alone. Correlation analysis showed that urinary cotinine concentrations were positively associated with urinary excretion of thioether and DGA, indicating that cigarette smoke induces the activity of hepatic enzymes and acts as a source of substrates metabolised through a thioether pathway. In conclusion it seems that exposure to petroleum vapour causes increased sister chromatid exchange in circulating lymphocytes of cigarette smokers, possibly as a result of enhanced hepatic conversion of vapour components to reactive metabolites. Urinary thioether output does not clearly discriminate between workers exposed to different amounts of petroleum vapour at retail outlets.

Adolescent↗

The coupling of supercritical fluid chromatography and field ionization time-of-flight high-resolution mass spectrometry for rapid and quantitative analysis of petroleum middle distillates.

We report the first coupling of supercritical fluid chromatography (SFC) with field ionization time-of-flight high-resolution mass spectrometry (FI-ToF HRMS), in parallel with ultraviolet (UV) detection and flame ionization detection (FID), for rapid and quantitative analysis of petroleum middle distillates. SFC separates petroleum middle distillates into saturates and 1- to 3-ring aromatics. FI generates molecular ions for hydrocarbon species eluted from the SFC. The high resolution and exact mass measurements by ToF mass spectrometry provide elemental compositions of the molecules in the petroleum product. The amounts of saturates and aromatic ring types were quantified using the parallel SFC-FID assisted by SFC-UV. With a proper carbon-number calibration, the detailed composition of the petroleum middle distillate was rapidly determined.

Journal Article↗

The environmental impact of petroleum on the environment.

Nigeria, a Western African country is particularly prone to the effects of oil pollution because it produces large quantities of crude oil (1.9 million barrels per day) for export. Results compiled by the Nigeria National Petroleum Corporation over a period of 15 years i.e. between 1976 to 1991 showed that there was a total of 2,976 spills, resulting in the release of 2 million barrels of crude oil into the Nigerian environment, with damaging health and ecological effects, particularly on the southern coastal communities of the country. The environmental and health impact of these spills is made worse by the inappropriate and inadequate environment and health impact assessment processes and policies in Nigeria. However proper environmental and health impact assessment processes of petroleum drilling and transport projects is a major factor helping in the prediction and reduction of the health and environmental impacts of petroleum on the local environment in western Australia. A comparison of the crude oil spills, and its associated environmental and health effects in Western Australia and Nigeria revealed that the rather devastating impact oil pollution in Nigeria is due, in part, to the poor environmental and health impact assessment policies and control procedures employed in Nigeria, particularly in the area of public participation and governmental control of petroleum projects.

Journal Article↗

Petroleum distillates suppress in vitro metabolic activation: higher [S-9] required in the Salmonella/microsome mutagenicity assay.

To determine if standard conditions used in the Salmonella/mammalian microsome mutagenicity assay could reliably screen complex petroleum samples, two high-boiling (700-1,070 degrees F) distillates and their separated aromatic fractions were tested. The initial mutagenic activities were inconsistent with the samples' known polyaromatic hydrocarbon (PAH) contents and observed potencies in a dermal carcinogenesis bioassay. A significant mutagenic response was observed only at S-9 concentrations 5 to 10 times higher than those used in the standard assay, supporting the use of elevated levels of S-9 in the Salmonella/microsome assay to assess the carcinogenic potential of petroleum-derived materials. All four samples masked the expected mutagenic activity of added PAHs (benzo[a]pyrene and perylene). Data suggested that petroleum distillates suppress the functional efficacy of the S-9; possible mechanisms are discussed.

Animals↗

Beyond TPH: health-based evaluation of petroleum hydrocarbon exposures.

The term "total petroleum hydrocarbons" (TPH) is a widely used, but loosely defined, parameter quantified by a number of different methodologies for expressing the aggregate amount of petroleum hydrocarbon compounds (PHCs) in a sample. Because of the shortcomings associated with comparing data from different methods, and the difficulty of assessing potential toxicities of complex mixtures of hydrocarbons, a new approach at more fully and explicitly defining the PHC composition of samples and predicting human noncancer health risks from those exposures has been developed. This new approach is the subject of this paper. This method can be used to perform site-specific risk assessments or to develop health-based cleanup standards for petroleum hydrocarbons. The technique divides the broad chemical classes of PHC (i.e., saturated versus unsaturated) into subgroups of compounds based on numbers of carbon atoms in the compounds within each subgroup. The mass of compounds in each subgroup is then translated into discrete estimates of health risk for specified exposure scenarios. The subgroups were identified from qualitative and quantitative changes in the nature of noncancer toxicities recorded in the literature. For saturated compounds, toxicity changes as carbon chain length increases (measured by numbers of carbon atoms). A "reference compound" was chosen for each range of compounds, usually because its toxicity was relatively well characterized. A published oral reference dose (RfD) was identified for these compounds, or in the absence of a published value, an oral dose-response value was developed from available toxicity information. For saturated PHCs (alkanes, cycloalkanes, and isoalkanes) the subgroups' reference compounds and assigned toxicity value used are C5 to C8 (n-hexane, 0.06 mg/kg/day); C9 to C18 (n-nonane, 0.6 mg/kg/day); and C19 to C32 (eicosane, 6.0 mg/kg/day). For unsaturated compounds (aromatics), one reference RfD was identified for all compounds: C9 through C32 (pyrene, 0.03 mg/kg/day). Dependent upon the analytical technique used for separation of compounds, the unsaturated alkenes may be grouped and subsequently quantified with either the saturate or unsaturate groups. The implications of possible association with either group and contributions to risk estimates are probably not significant. Alkenes make up a small fraction of most fuel products, and they bear structural similarity to the alkanes and are not particularly toxicologically active. If grouped analytically with the aromatics the alkene contribution to toxicity estimates would likely be minor and not be an underestimate of its true toxicity. The mass of PHC in each segment of a chromatogram is quantified and converted to a medium-specific concentration which is then entered into standard medium intake equations to arrive at a daily dose of PHC. This dose is then used with the toxicity value identified for the particular segment of the chromatogram to derive a hazard quotient. The quotients can then be summed across fractions to yield a total hazard index. The noncancer health risks from the aromatics benzene, toluene, and xylenes are evaluated separately using standard risk assessment techniques.

Alkanes↗

The effect of plants on the degradation and toxicity of petroleum contaminants in soil: a field assessment.

A field project located at the US Naval Base at Port Hueneme, California was designed to evaluate changes in contaminant concentrations and toxicity during phytoremediation. Vegetated plots were established in petroleum (diesel and heavy oil) contaminated soil and were evaluated over a two-year period. Plant species were chosen based on initial germination studies and included native California grasses. The toxicity of the impacted soil in vegetated and unvegetated plots was evaluated using Microtox, earthworm, and seed germination assays. The reduction of toxicity was affected more by contaminant aging than the establishment of plants. However, total petroleum hydrocarbon concentrations were lower by the end of the study in the vegetated plots when compared to the unvegetated soil. Although phytoremediation is an effective approach for cleaning-up of petroleum contaminated soil, a long-term management plan is required for significant reductions in contaminant concentrations.

Bacteria↗

Petroleum pollution in the Gulf of Mexico and Caribbean Sea.

In 1976, IOC-UNESCO and UNEP convened a meeting in Port of Spain to analyze the marine pollution problems in the region, noting that petroleum pollution was of regionwide concern and recommended initiating a research and monitoring program to determine the severity of the problem and monitor its effects. The Wider Caribbean is potentially one of the largest oil-producing areas in the world. Major production sites include Louisiana and Texas in the U.S.; the Bay of Campeche, Mexico; Lake Maracaibo, Venezuela; and the Gulf of Paria, Trinidad. All these are classified as high-risk production accident zones. Main sources of petroleum pollution in the Wider Caribbean are production, exploitation, transportation, urban and municipal discharges, refining and chemical wastes, normal loading and unloading operations, and accidental spills. About 5 million barrels of crude oil are transported daily in the Caribbean, thus generating an intense tanker traffic. It has been estimated that oil discharges from tank washings within the Wider Caribbean could be as high as 7 million barrels/yr. The results of the Caribbean Pollution Regional Program (CARIPOL) conducted between 1980 and 1987 pointed out that significant levels of petroleum pollution exist throughout the Wider Caribbean, including serious tar contamination of windward exposed beaches, high levels of floating tar within the major current systems, and very high levels of dissolved and dispersed hydrocarbons in surface waters. Major adverse effects of this type of pollution include: high tar levels on many beaches that either prevent their recreational use or require very expensive cleanup operations, distress and death for marine life, and responses in the enzyme systems of marine organisms that have been correlated with declines in reproductive success. Finally, the presence of polycyclic aromatic hydrocarbons (PAHs) in tissues of important economic species has been reported, creating a risk for public health because of the potential carcinogenic effects.

Accidents, Occupational↗

32P-postlabeling of DNA adducts arising from complex mixtures: HPLC versus TLC separation applied to adducts from petroleum products.

The carcinogenicity of petroleum products is mainly due to their content of polycyclic aromatic compounds (PACs). These compounds may be activated metabolically and react with DNA to form DNA adducts, which is a critical event in the initiation of cancer. One of the most common techniques for analyzing DNA adducts is (32)P-postlabeling. The chromatographic method often used has been (32)P-TLC (thin-layer chromatography), but the more recently developed (32)P-HPLC (high-performance liquid chromatography) method has shown advantages. The aim of this study was to test the hypothesis that the (32)P-HPLC method has a better ability of detecting DNA adducts derived from petroleum products than (32)P-TLC. It was found that some DNA adducts migrated from the application point in (32)P-TLC in such a way that it is doubtful if they could be detected and quantified properly. It was also found that, when using (32)P-HPLC, it is possible to use the same protocol for substances with a wide variety of DNA adduct forming potential, whereas (32)P-TLC needs to be optimized regarding time of exposure and/or the amount of DNA applied. Further, a pattern of recognition in (32)P-HPLC enables a selective assessment of DNA adducts derived from complex mixtures whereas (32)P-TLC is very limited when analyzing complex mixtures due to poor resolution. With more knowledge about the properties of the most mutagenic DNA adducts in HPLC, it could be possible to know also which pattern corresponds to a mutagenic or carcinogenic oil. Consequently, (32)P-HPLC is a good alternative when assessing the genotoxicity of petroleum products.

Chromatography, High Pressure Liquid↗

Influence of petroleum hydrocarbon contamination on microalgae and microbial activities in a long-term contaminated soil.

Petroleum hydrocarbons are widespread environmental pollutants. Although biodegradation of petroleum hydrocarbons has been the subject of numerous investigations, information on their toxicity to microorganisms in soil is limited, with virtually no work conducted on soil algae. We carried out a screening experiment for total petroleum hydrocarbons (TPH) and their toxicity to soil algal populations, microbial biomass, and soil enzymes (dehydrogenase and urease) in a long-term TPH-polluted site with reference to an adjacent unpolluted site. Microbial biomass, soil enzyme activity, and microalgae declined in medium to high-level (5,200-21,430 mg kg(-1) soil) TPH-polluted soils, whereas low-level (<2,120 mg kg(-1) soil) pollution stimulated the algal populations and showed no effect on microbial biomass and enzymes. However, inhibition of all the tested parameters was more severe in soil considered to have medium-level pollution than in soils that were highly polluted. This result could not be explained by chemical analysis alone. Of particular interest was an observed shift in the species composition of algae in polluted soils with elimination of sensitive species in the medium to high polluted soils. Also, an algal growth inhibition test carried out using aqueous eluates prepared from polluted soils supported these results. Given the sensitivity of algae to synthetic pollutants, alteration in the algal species composition can serve as a useful bioindicator of pollution. The results of this experiment suggest that chemical analysis alone is not adequate for toxicological estimations and should be used in conjunction with bioassays. Furthermore, changes in species composition of algae proved to be more sensitive than microbial biomass and soil enzyme activity measurements.

Biomass↗