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Mutagenicity and chemical analysis of aliphatic and aromatic fractions of Prudhoe Bay crude oil and fuel oil no. 2.

The mutagenicity of aliphatic (fraction 1), one- to three-ring aromatic (fraction 3), and four- to five-ring aromatic (fraction 4) fractions of Prudhoe Bay crude oil and of a fuel oil no. 2 was investigated with a battery of in vitro mammalian and microbial assays. Chemical analysis of the fractions was carried out by gas capillary chromatography-mass spectrometry. In the Salmonella/mammalian microsome test, both first fractions were negative, while fraction 3 of Prudhoe Bay crude and fraction 4 of both oils exhibited mutagenic activity with metabolic activation. In Chinese hamster ovary cells, all of the aromatic fractions, in the presence of metabolic activation, caused a significant increase in the number of sister chromatid exchanges. Chromosome aberrations were not caused by any of the aromatic fractions; however, fraction 3 of fuel oil no. 2 induced endoreduplication in the presence of metabolic activation.

Animals

Comparison of biodegradability of crude and fuel oils.

Crude and fuel oils were compared for ability to support growth of a mixed population of estuarine bacteria. A total of four oils, two crude and two fuel oils, were examined. It was found that each of the oils supported a unique population of bacteria and yeasts, with respect to generic composition. Low-sulfur, high-saturate, South Louisiana crude oil was found to be highly susceptible to degradation. In contrast, the dense, high-sulfur, high-aromatic, Bunker C fuel oil was strongly refractory to microbial degradation.

Alkanes

Effect of moisture content on the microbial activity in JP-5 fuel oil.

The water solubility of JP-5 fuel oil was found to be proportional to the reciprocal of absolute temperature from 0 degree C to 60 degrees C. Water in the fuel oil would become condensed once the temperature was shifted from a high temperature to low temperature. During the storage, condensed water was precipitated in the bottom of the tank. Both the static and the dynamic dehumidification processes with molecular sieve could reduce the moisture content of fuel oil to less than 5 ppm. However, pre-dried fuel oil had a mildly hydroscopic phenomenon at relatively high humidity condition. The spores of contaminated microbes could survive in the fuel oil with 5 to 80 ppm of moisture content. High moisture content of fuel oil was not favorable to the spore survival. Penicillium sp. could survive in the fuel oil longer than Cladosporium resinae.

Cladosporium

Effects of chronic ingestion of No. 2 fuel oil on mallard ducklings.

No. 2 fuel oil was fed to mallard (Anas platyrhynchos) ducklings in concentrations of 0.5 and 5.0% of the diet from hatching to 18 wk of age to assess the effects of chronic oil ingestion during early development. Five growth parameters (body weight, wing length, ninth primary length, tarsal length, and bill length) were depressed in birds receiving a diet containing 5% fuel oil. There was no oil-related mortality. The 5% fuel oil diet impaired avoidance behavior of 9-d-old mallard ducklings compared with controls or ducklings fed 0.5% oil. Open-field activity was greatly increased in 16-wk-old ducklings fed 5.0% oil. Liver hypertrophy and splenic atrophy were gross evidences of pathological effects in birds on the 5.0% oil diet. More subtle effects included biochemical lesions that resulted in the elevation of plasma alanine aminotransferase and ornithine carbamoyltransferase activity.

Alanine Transaminase

Upper airway response in workers exposed to fuel oil ash: nasal lavage analysis.

OBJECTIVES: Among other constituents, fuel oil ash contains vanadium pentoxide, a known respiratory irritant. Exposure to ambient vanadium pentoxide dust has been shown to produce irritation of the eyes, nose, and throat. The usefulness of nasal lavage in detecting an inflammatory response to exposure to fuel oil ash among 37 boilermakers and utility workers was investigated. METHODS: A baseline lavage was performed on the morning of the first day back to work after an average of 114 days away from work (range 36 hours to 1737 days). A lavage was performed after exposure on the morning three days after the baseline lavage. Exposure to respirable particulate matter of diameter < or = 10 microns (PM10) and respirable vanadium dust were estimated with daily work diaries and a personal sampling device for respirable particulates. These estimates were made for each subject on each workday during the three days between lavages. For each subject, the adjusted change in polymorphonuclear cells was calculated by dividing the change in polymorphonuclear cell counts by the average of the counts before and after exposure. The association between the adjusted polymorphonuclear cell counts and exposure was assessed with multiple linear regression, adjusted for age and current smoking. RESULTS: Personal sampling (one to 10 hour time weighted average) showed a range of PM10 concentrations of 50 to 4510 micrograms/m3, and respirable vanadium dust concentration of 0.10 to 139 micrograms/m3. In smokers the adjusted polymorphonuclear cell count was not significantly different from zero (-0.1%, P > 0.5), but in nonsmokers it was significantly greater than zero (+50%, P < 0.05). In both non-smokers and smokers, there was considerable variability in adjusted polymorphonuclear cell counts and a dose-response relation between these adjusted cell counts and either PM10 or respirable vanadium dust exposure could not be found. CONCLUSION: A significant increase in polymorphonuclear cells in non-smokers but not smokers was found. This suggests that in non-smokers, exposure to fuel oil ash is associated with upper airway inflammation manifested as increased polymorphonuclear cell counts. The lack of an increase in polymorphonuclear cells in smokers may reflect either a diminished inflammatory response or may indicate that smoking masks the effect of exposure to fuel oil ash.

Adult

[The influence of diesel fuel oil on the number of bacteria, fungi, actinomycetes and soil microbial biomass].

In laboratory conditions the influence of Diesel fuel oil on the total number bacteria, fungi, actinomycetes and content of microbial biomass in soil was studied. The fuel oil was applied at dosage of 1, 2 and 5 g/100 g of soil. Samples of soil were analysed 1, 7, 14, 21 and 150 days after oil applied. Microbiological analyses involved determinations of the total number of bacteria and actinomycetes (on Bunta-Roviry medium) and fungi (on Martin's agar) by the plate method. The content of microbial biomass in soil was determined using the physiological method of Anderson and Domsch. Diesel fuel oil application caused the changes of total number of investigated groups of soil microorganisms. These results were confirmed by the investigated groups of soil microorganisms. These results were confirmed by the investigation on microbial biomass in soil. In the most of cases the fuel oil was the most stimulated for the number of bacteria and the most inhibited for actinomycetes in comparison with soil without oil (control). From soil with Diesel fuel oil addition Pseudomonas, Arhtrobacter, Bacillus and Penicillium, Aspergillus, Fusarium, Trichoderma strains were isolated. The possibility of the fuel oil as source of carbon utilization by these strains will be investigate.

Actinomycetales

Toxicity and sublethal effects of No. 2 fuel oil on the supralittoral isopod Lygia exotica.

1. No. 2 fuel oil was of relatively low toxicity to the intertidal isopod Lygia exotica as indicated by the TLm values of over 100% for the WSF and 73 ppm at 24 and 48 hours and 36.5 ppm at 96 hours for the OWD. 2. Respiration was not significantly affected by short term exposure to several concentrations of No. 2 fuel oil prepared as either a WSF or OWD. 3. Lygia contamined by a spill of No. 2 fuel oil and Bunker C residual oil contained high concentrations of dibenzothiophenes. It is not known whether the dibenzothiophenes were accumulated by the Lygia tissues or absorbed to the exoskeleton. Therefore, the high mortality of Lygia following the spill cannot yet be attributed to the dibenzothiophenes.

Animals

Toxic and genetic effects of fuel oil photoproducts and three hydroperoxides in Saccharomyces cerevisiae.

Phototransformation of no. 2 fuel oil by UV irradiation at wavelengths designed to simulate sunlight resulted in the formation of products toxic to the yeast Saccharomyces cerevisiae. Increasing the time of irradiation of the fuel oil samples increased the toxicity. Fuel oil that had been irradiated for 12 or 24 h was convertagenic to the yeast strain D4. The toxicity and genetic activity of these samples could be removed by treatment with thiacyclohexane. It is thought that hydroperoxides are the primary photoproducts responsible for these biological effects. Of three hydroperoxides tested, tert-butyl was convertagenic and cumene and tetralin were not. However, all three hydroperoxides were toxic to yeast.

Fuel Oils

Determination of total sulphur in fuel oils by molecular emission cavity analysis.

Molecular emission cavity analysis (MECA) is used for the determination of total sulphur in fuel oils. The sulphur is reduced by heating the sample with Devarda's alloy in an HCl medium, with subsequent liberation of H2S and measurement of the chemiluminescent S2 emission intensity in the MECA cavity. The total sulphur recovery from different sulphur-containing compounds and fuel oils is always ca. 100%. The precision and accuracy are good. Applications to certified and commercial fuel oils are discussed.

Fuel Oils

FLIT-MLO and No. 2 fuel oil: effects of aerosol applications to mallard eggs on hatchability and behavior of ducklings.

FLIT-MLO and No. 2 fuel oil are sprayed on wetlands for mosquito control during spring and summer. In one experiment to assess the effects of the spraying on birds, mallard eggs were sprayed with amounts of No. 2 fuel oil equivalent to 2.34, 4.67, or 18.70 liters/ha or FLIT-MLO equivalent to 9.35, 46.75, or 140.25 liters/ha on Day 6 of incubation. In a second experiment, mallard eggs were sprayed with 9.35, 46.75, or 140.25 liters/ha of FLIT-MLO on Days 3, 6, 12, or 18 of incubation. Hatchability of eggs sprayed with the highest treatment level of each substance was significantly lower than that of controls for the first experiment. Hatchability of eggs sprayed with FLIT-MLO in the second experiment was never significantly lower than that of controls. Ducklings from the first experiment, 36-48 hr old, were cold stressed for 1 hr at 8 degrees C and then immediately tested for their ability to respond to a fright stimulus. Ducklings from the group of eggs sprayed with 140.25 liters/ha of FLIT-MLO ran a significantly shorter distance from the fright stimulus than did controls. The effects of the heaviest exposure to FLIT-MLO (140.25 liters/ha) on egg hatchability and behavior of newly hatched young are uncertain because of the contradictory results for hatching success in the two experiments. However, normal applications of FLIT-MLO (9.35-46.75 liters/ha) or No. 2 fuel oil (2.34-4.67 liters/ha) do not appear to pose a threat to the embryos of breeding birds.

Animals

Effects of No. 2 fuel oil on hatchability of marine and estuarine bird eggs.

Eggs of Louisiana herons, sandwich terns, and laughing gulls were oiled with either 0, 5, or 20 microliter of No. 2 fuel oil in the field and in the laboratory. After 5 days of natural incubation, field-oiled and control eggs were opened and embryonic mortality was determined. No. 2 fuel oil produced 61% mortality in Louisiana heron eggs, 56% in sandwich tern eggs, and 83% in laughing gull eggs. Hatching success of artificially incubated, oiled eggs appeared to be lower than in control eggs. However, stress during shipment to the laboratory and problems within the incubator probably contributed to reduced hatchability in both groups.

Animals

Chemical and toxicological characterization of residential oil burner emissions: I. Yields and chemical characterization of extractables from combustion of No. 2 fuel oil at different Bacharach Smoke Numbers and firing cycles.

Particulates and complex organic mixtures were sampled from the exhaust of a flame retention head residential oil burner combusting No. 2 fuel oil at three firing conditions: continuous at Bacharach Smoke No. 1, and cyclic (5 min on, 10 min off) at Smoke Nos. 1 and 5. The complex mixtures were recovered by successive Soxhlet extraction of filtered particulates and XAD-2 sorbent resin with methylene chloride (DCM) and then methanol (MeOH). Bacterial mutagenicity [see Paper II (8)] was found in the DCM extractables. Samples of DCM extracts from the two cyclic firing conditions and of the raw fuel were separated by gravity column chromatography on alumina. The resulting fractions were further characterized by a range of instrumental methods. Average yields of both unextracted particulates and of DCM extractables, normalized to a basis of per unit weight of fuel fired, were lower for continuous firing than for cyclic firing. For cyclic firing, decreasing the smoke number lowered the particulates emissions but only slightly reduced the average yield of DCM extractables. These and similar observations, here reported for two other oil burners, show that adjusting the burner to a lower smoke number has little effect on, or may actually increase, emissions of organic extractables of potential public health interest. Modifications of the burner firing cycle aimed at approaching continuous operation offer promise for reducing the amount of complex organic emissions. Unburned fuel accounted for roughly half of the DCM extractables from cyclic firing of the flame retention head burner at high and low smoke number. Large (i.e., greater than 3 ring) polycyclic aromatic hydrocarbons (PAH) were not observed in the DCM extractables from cyclic firing. However, nitroaromatics, typified by alkylated nitronaphthalenes, alkyl-nitrobiphenyls, and alkyl-nitrophenanthrenes were found in a minor subfraction containing a significant portion of the total mutagenic activity of the cyclic low smoke samples (8). Oxygen-containing PAH, typified by phenalene-1-one and its alkyl derivatives, are important mutagens from cyclic firing at high smoke conditions. Thus, oil burner effluents differ markedly from those of several other combustors, including the automotive diesel engine, where multiring PAH, typified by fluoranthene and alkylated phenanthrenes, account for a significant portion of the effluent mutagenicity. Implications for combustion and emissions source identification are discussed.

Air Pollutants

Determination of polycyclic aromatic hydrocarbons in diesel exhaust particulate matter and diesel fuel oil.

Clean-up procedures were developed for a method for determining the amount of polycyclic aromatic hydrocarbons (PAHs) in diesel exhaust particulate matter and in diesel fuel oils using reversed-phase high-performance liquid chromatography (HPLC). They were based mainly on the elimination of insoluble matter and aliphatic compounds that affect the performance of HPLC, from the dichloromethane extracts of particulate matter or from oils, with the aid of a disposable preparation column containing reversed-phase packings (Sep-Pak C18). Using these procedures, it is possible to detect 1 ng of benzo(a)pyrene in 30 mg of particulate matter with more than a 97% recovery or 0.5 ng in 50 microliters of oil with 91% recovery. Examples of analyses are given for particulate matter emitted from a diesel test engine and for diesel fuel oils, such as gas oil, residual oil and coal-liquefied oil.

Chromatography, High Pressure Liquid

Petroleum hydrocarbon resistance in the marine worm Neanthes arenaceodentata (polychaeta: annelida), induced by chronic exposure to No. 2 fuel oil.

1. Three successive generations of the marine polychaetous annelid Neanthes arenaceodentata taken from a laboratory population, were continuously exposed to one of three sublethal concentrations of No. 2 Fuel Oil water-soluble-fraction (WSFs). During each generation larvae, juvenile, and immature adult polychaetes were challenged with acute (96 hr) doses of No. 2 Fuel Oil or south Louisiana crude oil WSF to test their sensitivity to petroleum hydrocarbons (PHCs). 2. Larvae from all 3 generations, at all exposure concentrations, were no different from control (susceptible) larvae in their sensitivity to the two test oils. F1, F2, and F3 adults exhibited equally increased PHC resistance (X2) compared to control adults. 3. The only evidence of increased resistance beyond that observed in F1 animals was seen in results of bioassays with juvenile worms, wherein PHC resistance increased from slightly below control levels in F1 juveniles to slightly above control tolerance among F3 juveniles. 4. With the exception of F1 worms, removal from chronic exposure 7 or 14 days prior to challenge did not result in termination or reduction of resistance, implicating a genetic mechanism behind PHC resistance in N. arenaceodentata. 5. F3 resistant and unexposed control polychaetes accumulated, metabolized, and excreted a key diaromatic PHC (naphthalene-14C) in quantitatively identical fashion. Mechanisms responsible for resistance appeared unrelated to external permeability and/or excretion rates.

Animals

Developmental toxicity of EDS recycle solvent and fuel oil.

Direct coal liquefaction is one of several technologies currently under development as alternative means to produce liquid fuels. Relatively high levels of polycyclic aromatic hydrocarbons are present in distillate fractions boiling above approximately 370 degrees C. Coal-derived liquids containing substantial amounts of material from this boiling range were genotoxic in in vitro tests and carcinogenic in mouse skin. Some of the liquids were also teratogenic in rodents. The present report describes studies which assessed the potential effects of 2 coal-derived liquids, recycle solvent (nominal boiling range 200-427 degrees C) and an experimental industrial fuel oil (nominal boiling range 204-538 degrees C) on prenatal development in the rat. The test materials were produced by the EDS direct coal liquefaction process and contained substantial amounts of material boiling above 370 degrees C. Test materials were administered by gavage to pregnant female Sprague-Dawley rats from days 6 to 19 of gestation (G). Animals were sacrificed on day 20G and the uterine contents were removed and examined. Results of both studies were similar. The number of live fetuses declined in a dose-related manner, and there was evidence of intrauterine growth retardation in fetuses which survived to day 20G. Statistically significant effects were noted at doses which did not appear to be maternally toxic. The frequency of malformation was not significantly elevated in either study; however, a thorough evaluation of this endpoint was precluded by embryo lethality at the high doses. It was apparent that both of the EDS liquids examined affected prenatal survival and growth. However, in contrast to studies of other coal-derived liquids, there was no evidence of teratogenic effects at non-toxic doses.

Administration, Oral

Persistence and biodegradation of spilled residual fuel oil on an estuarine beach.

The enrichment of hydrocarbon-degrading bacteria and the persistence of petroleum hydrocarbons on an estuarine beach after a spill of residual fuel oil on 11 April 1973 in Upper Narragansett Bay, R.I. was investigated. A rapid enrichment occurred during days 4 to 16 after the oil spill and a significant population of hydrocarbon-degrading bacteria was maintained in the beach sand for at least a year. The concentration of petroleum hydrocarbons in the mid-tide area declined rapidly during the bacterial enrichment period, remained fairly constant throughout the summer, and then declined to a low concentration after 1 year. An increased concentration of branched and cyclic aliphatic hydrocarbons in the low-tide sediment 128 days after the spill suggested a migration of hydrocarbons during the summer. Hydrocarbon biodegradation was apparent during the winter months at a rate of less than 1 mug of hydrocarbon per g of dry sediment per day.

Alkanes

Gas chromatographic-mass spectrometric determination of aromatic hydrocarbon metabolites from livers of fish exposed to fuel oil.

Metabolites of several two- and three-ring aromatic hydrocarbons (AHs) have been found in livers of English sole exposed to No. 2 fuel oil. Four metabolites of the C2H5-naphthalenes, six of the C3H7-naphthalenes and one each of fluorene, phenanthrene and anthracene have been partially characterized and their concentrations, which ranged from 50 to 1100 ng/g, were determined. Metabolites were separated from the liver matrix using an automated extractor/concentrator. The resulting extract was then purified by high-performance liquid chromatography, and the metabolites were characterized and quantitated by gas chromatography-mass spectrometry.

Animals