Pulmonary function evaluation of cats after one year of exposure to diesel exhaust

Adult male, inbred, disease-free cats of uniform age and size were exposed eight hours per day, seven days per week to a 1 : 18 dilution of diesel exhaust emissions. After one year of exposure, the animals were removed from the chambers for measurement of lung volumes, forced expiratory flow rates, dynamic compliance and resistance, diffusing capacity, and nitrogen washout. No important changes in pulmonary function were detected with the exception of a decrease in closing volume (P < 0.05). The inability to detect decrements in pulmonary function may have been due to insufficient cocentration of exhaust, insufficient exposure length, or to the use of a species resistant to diesel exhaust. To test these possibilities, the cats are being exposed for an additional year, and another species, hamsters, are being exposed for future testing at exhaust dilutions of 1 : 18 and 1 : 9.     

Neurophysiological alterations due to diesel exhaust exposure during the neonatal life of the rat

This study was designed to assess the effects of diesel exhaust on the development of the nervous system in rats as measured by somatosensory and visual evoked potentials (SEPs and VEPs, respectively). SEPs, elicited by 1 mamp, 0.5 msec pulses delivered to the tibial nerve at the tibale, were recorded from the skull overlying the somatosensory projection area on days 28, 35, 42, and 49 postconception (pc). Analysis of the data for day 35 pc double pulse SEPs revealed significantly longer latencies for all peaks of the SEP in pups exposed to diesel exhaust. There were also differences indicating that the electrophysiological recoverability of the nervous system had been compromised in the diesel exhaust exposed group. On day 35 pc, the latencies of all VEP peaks occurring before 300 msec were greater in the exposed group. The differences, however, reached statistical significance only for the P2 peak of the response to the first of the paired stimuli. These data support the hypothesis that diesel exhaust influences the development of the nervous system.     

Lymphatic transport of inhaled diesel particles in the lungs of rats and guinea pigs exposed to diluted diesel exhaust

Factors influencing the disposition of the inhaled diesel particles were studied by analyzing the deposition of radioactively labelled diesel particles in the respiratory system, by determining the specific function of alveolar cellular mechanisms in the primary defense against inhaled particles and by identifying the important role of the lymphatic system in the lung clearance of experimental animals exposed to diluted emissions from a diesel engine. Radioactive ¹³¹Barium was used as a tracer of diesel particles and the deposition efficiency was determined to be 15%±6% of the inhaled dose in the Fischer 344 rat strain. The number of cells obtained by bronchial lavage increased significantly after a prolonged exposure to a concentration of 1500 μg/m³ of diesel particles. The increased cell number was more than twofold, contained two distinct cell populations (alveolar macrophages and neutrophils) and represented a reactive mobilization of the defense mechanisms in the organism. Light microscopy studies investigated the role of lymphatic transport of the particulate matter and revealed that the peribrochial and perivascular aggregates of lymphoid tissue contained diesel particles even after short exposure periods at low dose levels. With the increasing burden of particles in the respiratory system, the coloration of hilar and mediastinal lymph nodes continuously changed to gray and finally to dark black, depending upon the dose level and exposure. However, at all exposure levels, most of the diesel particles in the alveoli were phagocytized by an increased alveolar cellular defence and particle-containing macrophages were actively moving towards the mucociliary escalator or towards lymphatic channels leading to peribronchial lymphoid aggregates and bronchial or mediastinal lymph nodes. In the lymph nodes, alveolar macrophages containing diesel particles were found mostly in the afferent subcapsular lymphatic vessels and marginal sinuses. In the later stages, cellular structure disintegrated and large aggregates of particulate matter were dispersed throughout the medullary cords with increasing accumulation towards the hilus. It is concluded that the lymphoid aggregates and lymphatic nodes play an important role in sequestering diesel particles or particle-containing phagocytizing cells and provide a pathway, in addition to the mocociliary clearance for particulate removal from the deep pulmonary region.     

Preparation and characterization of diesel exhaust particles for biological experiments

Bioassays of diesel engine exhaust components are being conducted at IITRI to determine toxic and carcinogenic potentials of the exhaust. The bioassay method, intracheal instillation of saline suspensions of test materials in hamsters, requires preparation of stable suspensions of test materials. A method to prepare suspensions of whole particle diesel exhaust in saline has been developed. The diesel exhaust particle material was supplied to IITRI as a dry, loose powder by the U.S. EPA from a light duty diesel test engine. Preliminary characterizations of the powders indicated aggregation of exhaust particles had occurred both before and during capture on collection substrates. Flake-like sheets and hollow spheres of aggregated particles up to 150 μm in sie present in the powders. Therefore, the powder were ball-milled to geometric particle sizes more amenable to the animal administration technique to be employed. Grinding, suspension preparation and particle concentration assaying methods have been developed. Particle (geometric) size and morphological characterizations have also been performed on the as-received powders and prepared suspensions. A method to prepare emulsions (liquid-liquid suspensions) of the dichloromethane extracts of whole particle diesel exhaust has also been developed.     

Test of diesel exhaust emissions in the rat liver foci assay

In the initiation/promotion assay in rat liver, partial hepatectomy is used to enhance initiation, and a choline-devoid diet as promoter. The induction of carcinogenesis is determined by the focal appearance of gamma glutamyl transpeptidase (GGT) positive hepatocytes. We adopted this assay to diesel exhaust emission by performing a partial hepatectomy, and exposing the rats to either clean air or diesel exhaust emission. The rats were fed either a choline-devoid or a choline-supplemented diet for three or six months. The animals were sacrificed and liver sections stained for GGT were examined for the presence of foci of GGT(+) hepatocytes. The results indicate that diesel exhaust exposure does not result in a systemic dose of carcinogens sufficient to be detected in the liver foci assay.     

Pulmonary function testing of animals chronically exposed to diluted diesel exhaust for 267 days

Thw purpose of this work was to assess the potential effect that chronic inhalation of diesel exhaust may have on lung mechanics and lung volumes. Noninvasive pulmonary function tests have been conducted repeatedly on 25 rats exposed to diesel exhaust at a particulate concentration of 1500 μg/m³, for 20 h/day, days/week for 267 days. The same tests were conducted on 25 clean air control animals. When the data is normalized, there are no apparent functionally significant changes occurring in the lungs that may be attributed to the chronic inhalation of diesel exhaust.     

Modelling and experimental studies on the transfer of radionuclides to fruit

Although fruit is an important component of the diet, the extent to which it contributes to radiological exposure remains unclear, partially as a consequence of uncertainties in models and data used to assess transfer of radionuclides in the food chain. A Fruits Working Group operated as part of the IAEA BIOMASS (BIOsphere Modelling and ASSessment) programme from 1997 to 2000, with the aim of improving the robustness of the models that are used for radiological assessment. The Group completed a number of modelling and experimental activities including: (i) a review of experimental, field and modelling information on the transfer of radionuclides to fruit; (ii) discussion of recently completed or ongoing experimental studies; (iii) development of a database on the transfer of radionuclides to fruit; (iv) development of a conceptual model for fruit and (v) two model intercomparison studies and a model validation study. The Group achieved significant advances in understanding the processes involved in transfer of radionuclides to fruit. The work demonstrated that further experimental and modelling studies are required to ensure that the current generation of models can be applied to a wide range of scenarios.     

Salmonella/microsome mutagenicity assays of exhaust from diesel and gasoline powered motor vehicles

Motor vehicle exhaust from prechamber injection diesel and gasoline powered passenger cars, sampled during US FTP 1973 test cycles and comprising both particulate matter and compounds condensable at ambient temperature, has been assayed for mutagenicity in the Salmonella/microsome test. Mutagenic components were to a large extent active in the absence of the mammalian microsomal preparation. The mutagenicity of both particulate matter and condensate from diesel exhaust and condensate from gasoline exhaust was decreased in the presence of the microsomal preparation whereas the mutagenicity of particulate matter from gasoline exhaust was enhanced by microsomal activation. A comparison between the investigated diesel and gasoline exhaust samples shows that the mutagenic effect in the Salmonella test of the diesel exhaust is more than ten times higher than that of the gasoline exhaust. Fractionation with respect to polarity indicates that the mutagenic components mainly are distributed in neutral aliphatic, aromatic, and oxygenated fractions. Tests for mutagenic monofunctional nitroarenes by an anaerobic assay indicate that such compounds at most are marginally present in the exhaust samples as compared with their presence in airborne particulate matter collected in an urban environment.     

Benzo[a]pyrene metabolism in mice exposed to diesel exhaust: II. Metabolism and excretion

In this study we examined the effect of diesel exhaust (DE) exposure on in vivo metabolism of benzo[a]pyrene (BaP). DE-exposed and unexposed A/Jax mice of group B were instilled intratracheally with ³H-BaP. At each time point of 2, 24, and 168 h after instillation five mice were killed and the liver, lungs, and testes were removed and frozen. Aliquots of the organs were homogenized in 2 ml water and each received 3 volumes of cold ethanol. Radioactivity in supernatant and precipitate was measured. The supernatant extracts were subjected to HPLC analysis on ALOX-T and on Zorbax ODS. The ALOX-T method was a modification of Autrup's procedure for conjugate assay (Biochem. Pharmacol.28, 1727, 1979). Fractions were (a) free BaP; (b) nonconjugated primary metabolites; (c) sulfate conjugates; (d) glucuronides, glutathiones, and other conjugates. By 2 h after instillation primary metabolites were found in liver and lung, but very little was conjugated. The unconjugated BaP was mainly in the form of free BaP and phenolic metabolite(s). The lungs of DE-exposed mice had less capacity to dispose of “bound” BaP 1 week after instillation.     

Benzo[a]pyrene metabolism in mice exposed to diesel exhaust: I. Uptake and distribution

In this study we examined the effect of diesel exhaust (DE) exposure on the disposition of a typical polycyclic aromatic hydrocarbon. DE-exposed and nonexposed A/Jax mice were divided into three groups and each mouse instilled intratracheally with benzo[a]pyrene (BaP). One group (A) received ¹⁴C-BaP, and at intervals of 2, 24, and 168 h, three mice from the group were killed and quick frozen for whole body autoradiography. Sagittal sections were cut at 0.5 mm intervals and autoradiograms prepared. Adjacent sections were studied so that radioactive areas were matched to specific organs. The second group (B) received ³H-BaP and at 2, 24, and 168 h these mice were killed. Livers, lungs, and testes were weighed and frozen. From these tissues metabolites were analyzed; these data are reported in the next paper. Histofluorescent examination of tissues from mice instilled with nonradioactive BaP (group C) confirmed that BaP was present in the lung. The autoradiography data are the basis for elucidating the BaP distribution in the mouse. Within 2 h after instillation radioactivity was detected in the entire animal, with most in lungs, liver, and GI tract. By 24 h after instillation considerable radioactivity had redistributed to the GI tract. At 168 h after instillation only a trace of label was found in the GI mucosa.     

Seasonal distribution and modeling of diesel particulate matter in the Southeast US

The fine and ultra fine size of diesel particulate mater (DPM) are of great health concern and significantly contribute to the overall cancer risk. In addition, diesel particles may contribute a warming effect on the planet's climate. The composition of these particles is composed principally of elemental carbon (EC) with adsorbed organic compounds, sulfate, nitrate, ammonia, metals, and other trace elements. The purpose of this study was to depict the seasonality and modeling of particulate matter in the Southeastern US produced by the diesel fueled sources (DFSs). The modeling results came from four one-month cases including March, June, September, and December to represent different seasons in 2003 by linking Models-3/CMAQ and SMOKE. The 1999 National Emissions Inventory Version 3 (NEI99) was used in this analysis for point, area, and non-road sources, whereas the National Mobile Inventory Model (NMIM) was used to create the on-road emissions. Three urban areas, Atlanta, Birmingham, and Nashville were selected to analyze the DPM emissions and concentrations. Even though the model performance was not very strong, it could be considered satisfactory to conduct seasonal distribution analysis for DPM. Important hourly DPM seasonality was observed in each city, of which higher values occurred at the morning traffic rush hours. The EC contributions of primary DPM were similar for all three sites (~ 74%). The results showed that there is no significant daily seasonality of DPM contribution to PM_2.5 for any of these three cities in 2003. The annual DPM contribution to total PM_2.5 for Atlanta, Nashville, and Birmingham were 3.7%, 2.5%, and 2.2%, respectively.     

Mutagenic activity of diesel emission particulate extracts and isolation of the mutagenic fractions

Mutagenic activity in diesel emission particulate extracts was detected by the Salmonella typhimurium/microsome assay. Direct-acting mutagens as well as promutagens requiring metabolic activation were detected. The extracts were fractionated into acidic, basic, and neutral fractions, and the neutral fraction was chromatographed into seven subfractions. Differences in the mutagenic potency of these fractions and subfractions were determined by the Salmonella assay. Fractions containing as yet unidentified compounds, but not polynuclear aromatic hydrocarbons, were found to make a major contribution to mutagenic activity of the extracts.     

Levels of household particulate matter and environmental tobacco smoke exposure in the first year of life for a cohort at risk for asthma in urban Syracuse, NY

The Syracuse, NY, AUDIT (Assessment of Urban Dwellings for Indoor Toxics) study was designed to quantify asthma agent levels in the inner-city homes of a birth cohort whose mothers had a diagnosis of asthma. Risk of exposure to particulate matter (PM), particle number and tobacco smoke was assessed in 103 infants' homes. Repeat measurements were made in 44% of the homes. Infants also were examined on a quarterly basis during the first year of life to monitor their respiratory health and urine cotinine levels. Overall geometric mean (GM) values for PM(2.5) of 21.2 μg/m(3) and for PM(10) of 31.8 μg/m(3) were recorded in homes at visit 1. GM values for PM(2.5) and PM(10) in smoking homes were higher at 26.3 and 37.7 μg/m(3), while values in non-smoking homes were 12.7 and 21.2 μg/m(3) respectively. Fifty-four percent of mothers (55/103) smoked at some point in pregnancy (39% smoked throughout pregnancy). Environmental tobacco smoke (ETS) exposure occurred in 68% of homes during the infants' first year. Significant to this study was the size- and time-resolved monitoring of PM at 140 home visits and the classification of PM count data. PM number counts ranged from continuously low levels (little indoor activity) to continuously high counts (constant indoor activity), and recorded apparent instances of prolonged repeated cigarette smoking. Wheezing in the first year of life was recorded for 38% of the infants (39/103). Adjusted logistic regression modeling demonstrated that elevated levels of indoor PM(2.5) (≥ 15 μg/m(3)) were a significant risk factor for infant wheezing after controlling for infant gender, mothers' age and education level, season of home visit and presence of carpeting (OR 4.21; 95% CI 1.36-13.03; p=0.013). An elevated level of the nicotine metabolite cotinine in infant urine also was associated with infant wheezing after adjusting for infant gender, mothers' age and education level (OR 5.10; 95% CI 0.96-27.24; p=0.057). ETS exposure was pervasive in the AUDIT cohort and a risk for developing infants in this urban population.     

Soil-ecological conditions of the zone of salt waste dumping and plant adaptation to them

Technogenic surface formations near salt waste dumps are subject to salinization by sodium chloride. Synanthropic representatives of local flora that have adapted to salinization are characterized by increased concentrations of salinizing ions; redistribution of chlorides, with their predominant accumulation in above-ground organs; selective absorption with respect to sodium, chloride, and potassium ions; and decreased activity of hydrogen ions in cells.     

Regulatory options for the development of health and environmental testing of diesel fuels and fuel additives

This paper discusses various options for regulations of fuels and fuel additives. Four options of evaluation of combustion products are described. These are disregarding combustion products, complete analysis, testing of exhaust mixtures, and partial analysis. Tests are adapted from regulations of EPA for evaluation of combustion products.     

The co-application of earthworms (Dendrobaena veneta) and compost to increase hydrocarbon losses from diesel contaminated soils

The feasibility of using composted civic waste for the remediation of a soil contaminated with petroleum hydrocarbons (extractable petroleum hydrocarbons (EPH) 10+/-1.8 g kg(-1) and total 16 USEPA PAH 1.62+/-0.5 g kg(-1)) was assessed. The effects of compost to soil ratio, in combination with and without earthworm presence (Dendrobaena veneta), upon the loss of contaminants were determined for EPH (GC-FID) and PAH (GC-MS), respectively. Increasing the ratio of compost substrate to hydrocarbon impacted soil (1:0.5, 1:1, 1:2 and 1:4 (soil:compost wt/wt)) in the absence of earthworms resulted in significantly (p<0.05) greater losses of both EPH and SigmaPAH after an 84 d incubation period, when compared to the soil only control. Where earthworms were present without compost, EPH losses were significantly (p<0.05) enhanced in the soil only treatment (33.4+/-5.3% residual) compared to the soil only control (54.4+/-5.3% residual). However, PAH loss in the soil only treatment (with-earthworm presence) were only slightly enhanced (65.3+/-9.3% residual), with respect to the soil only control (69.2+/-6.4% residual). Synergistic benefits of both earthworm and compost presence were most significant for PAHs (p<0.05), and less so for EPH. (14)C-respirometer studies, to establish catabolic competence in terms of microbial mineralisation of key hydrocarbons, complemented the hydrocarbon analysis.     

Mapping of the geogenic radon potential in France to improve radon risk management: methodology and first application to region Bourgogne

In order to improve regulatory tools for radon risk management in France, a harmonised methodology to derive a single map of the geogenic radon potential has been developed. This approach consists of determining the capacity of the geological units to produce radon and to facilitate its transfer to the atmosphere, based on the interpretation of existing geological data. This approach is firstly based on a classification of the geological units according to their uranium (U) content, to create a radon source potential map. This initial map is then improved by taking into account the main additional parameters, such as fault lines, which control the preferential pathways of radon through the ground and which can increase the radon levels in soils. The implementation of this methodology to the whole French territory is currently in progress. We present here the results obtained in one region (Bourgogne, Massif Central) which displays significant variations of the geogenic radon potential. The map obtained leads to a more precise zoning than the scale of the existing map of radon priority areas currently based solely on administrative boundaries.     

Mutagenic and carcinogenic potency of extracts of diesel and related environmental emissions: Summary and discussion of the results

The proposed conversion from gasoline powered automobiles to diesel powered vehicels has prompted the Environmental Protection Agency to evaluate the potential health effects associated with exposure to diesel emissions. At present, there is no direct epidemiological link between this exposure and human health. Therefore, a research program was constructed to compare the health effects associated with diesel emissions with those from other emission sources for which epidemiological information was available. The emission sources chosen were cigarette smoke, roofing tar, and coke oven. An additional comparative emission source which was a gasoline catalyst engine. Respirable particles from a variety of combustion sources have the potential of being carcinogenic and mutagenic. The objective of these studies was to determine the relative biological activity of the organic material adsorbed on these particles in both in vitro mutagenesis and in vitro and in vivo bioassays. The organic extracts from the following series of emission sources were quantitatively bioassayed in a matrix of tests for their carcinogenic and mutagenic activity: (1) a light-duty Oldsmobile diesel 350 engine; (2) a heavy-duty Caterpillar diesel engine; (3) a light-duty Nissan engine; (4) a Volkswagen Rabbit diesel engine; (5) cigarette smoke; (6) roofing tar; (7) coke oven; and (8) a gasoline catalyst Mustang. The test matrix consisted of the following bioassay: reverse mutation in Salmonella typhimurium; mitotic recombination in Saccharomyces cerevisiae; DNA damage in Syrian hamster embryo cells (SHE); sister chromatid exchange in CHO cells; gene mutation in L5178Y mouse lymphoma cells, Balb/c 3T3 mouse embryo fibroblasts and CHO cells; viral enhancement of SHE cells; oncogenic transformation in Balb/c 3T3 cells; and skin tumor initiation in SENCAR and C57 black mice. The results of this test matrix are discussed.     

Modelling the propagation of effects of chronic exposure to ionising radiation from individuals to populations

This study evaluated the potential effect of ionising radiation on population growth using simple population models and parameter values derived from chronic exposure experiments in two invertebrate species with contrasting life-history strategies. In the earthworm Eisenia fetida, models predicted increasing delay in population growth with increasing gamma dose rate (up to 0.6 generation times at 11 mGy h(-1)). Population extinction was predicted at 43 mGy h(-1). In the microcrustacean Daphnia magna, models predicted increasing delay in population growth with increasing alpha dose rate (up to 0.8 generation times at 15.0 mGy h(-1)), only after two successive generations were exposed. The study examined population effects of changes in different individual endpoints (including survival, number of offspring produced and time to first reproduction). Models showed that the two species did not respond equally to equivalent levels of change, the fast growing daphnids being more susceptible to reduction in fecundity or delay in reproduction than the slow growing earthworms. This suggested that susceptibility of a population to ionising radiation cannot be considered independent of the species' life history.     

Assessment of the reproductive capacity of <Emphasis Type="Italic">Pinus sylvestris</Emphasis> trees growing under conditions of chronic exposure to radionuclides of uranium and thorium series

The reproductive capacity of Scots pine (Pinus sylvestris L.) has been estimated in trees from the population growing in an area used to store tailings from uranium ore processing and radium production. Under such conditions, 12- to 15-year-old trees produce seeds of a quality inferior to that in areas with a normal background radiation level. As the weighted absorbed dose rate (at a weighting coefficient of 5) for generative organs increases from 0.002 ± 0.0003 to 0.344 ± 0.059 mGy/day, a proportional increase is observed in the proportion of empty seeds and the frequency of cytogenetic disturbances in the root meristem of seedlings, while the survival rate of seedlings decreases. The minimum absorbed dose rate leading to these changes in the pine population is 0.027 mGy/day, i.e., 12 times as high as the control value.