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Inhalation toxicity of sulfuryl fluoride in rats and rabbits.

The inhalation toxicity of the structural fumigant sulfuryl fluoride (SO2F2) was evaluated in rats and rabbits. Exposures for a preliminary 2-week study were 6 hr/day, 5 days/week, to 0, 100, 300, or 600 ppm SO2F2. Nine of ten rats at 600 ppm died or were moribund between the second and sixth exposures. Extensive kidney lesions were present in all rats exposed to 600 ppm, whereas only minimal renal changes were noted in rats at 300 ppm. Upper and lower respiratory tissues were inflamed in the single rat that survived the 2-week exposure to 600 ppm. Rabbits exposed to 600 ppm SO2F2 were hyperactive and one animal had a convulsion. Exposure to 300 or 600 ppm for 2 weeks resulted in vacuolation and/or malacia in the cerebrum of all rabbits and most of these rabbits also had moderate inflammation of nasal tissues; a few rabbits at 600 ppm had inflammation of the trachea or bronchi. A subsequent 13-week study evaluated rats and rabbits exposed to 0, 30, 100, or 300 ppm SO2F2 (337 ppm TWA for rabbits). Rabbits initially were exposed to a high concentration of 600 ppm; however, convulsions were noted in two animals after nine exposures and the concentration subsequently was reduced to 300 ppm. Vacuolation and/or malacia were observed in the cerebrum of all rabbits at the highest concentration; one rabbit exposed to 100 ppm also had cerebral vacuolation. Rabbits at the highest concentration, as well as one rabbit exposed to 100 ppm, had inflammation of the nasal tissues. Rats exposed to 300 ppm SO2F2 for 13 weeks had mottled incisor teeth, minimal renal effects, pulmonary histiocytosis, inflammation of nasal tissues, and cerebral vacuolation. Also, rats exposed to 100 ppm SO2F2 for 13 weeks had mottled teeth. Fluoride toxicity was suggested by mottled teeth in rats as well as elevation of serum fluoride levels in rats and rabbits exposed to SO2F2 for 13 weeks. Although repeated exposure of rats and rabbits to 100-600 ppm SO2F2 resulted in toxicity of the kidneys (rats only), brain, and respiratory system, no effects were detected in animals exposed to 30 ppm for 13 weeks.

Administration, Inhalation↗

The effects of inhalation exposure to sulfuryl fluoride on fetal development in rats and rabbits.

Sulfuryl fluoride is a fumigant insecticide used for soils and permanent structures. Pregnant Fischer 344 rats and New Zealand White rabbits were exposed to 0, 25, 75, or 225 ppm of sulfuryl fluoride vapor via inhalation for 6 hr/day on Days 6-15 and 6-18 of gestation, respectively. Among rats, maternal water consumption was increased in the 225 ppm exposure group, but there were no indications of embryotoxicity, fetotoxicity, or teratogenicity in any of the exposed groups. Among rabbits, maternal weight loss during the exposure period (Days 6-18) was observed in the 225 ppm group. Decreased fetal body weights, considered secondary to maternal weight loss, were also observed at 225 ppm. However, no evidence of embryotoxicity or teratogenicity was observed among rabbits in any exposure group. Thus, inhalation exposure to sulfuryl fluoride was not teratogenic in either rats or rabbits exposed to levels of up to 225 ppm, and fetotoxic effects (reduced body weights) were observed among fetal rabbits only at an exposure level that produced maternal weight loss.

Administration, Inhalation↗

Potential consequences of a fire in an insecticide storage facility.

Assumptions are identified and made in an attempt to model the acute human health risk associated with a hypothetical fire in an insecticide storage facility. The insecticides used in the model are endrin and dimethoate. The model indicates that persons residing a few hundred meters from the facility could suffer a variety of adverse effects, including possible death from contact with smoke from a prolonged, low-temperature fire. Knowledge of the special hazards of cool toxic smoke as well as current atmospheric conditions could be of use to fire fighters. That is, fire fighters might wisely choose to promote a fast, hot fire which would propel toxicants high into the atmosphere rather than risk local fumigation.

Animals↗

Disposition of [14C]methyl bromide in Fischer-344 rats after oral or intraperitoneal administration.

Methyl bromide is used as a disinfectant to fumigate soil. The intent of our study was to determine the disposition of methyl bromide following a single acute administration. Male Fischer-344 rats were given 250 mumol of [14C] methyl bromide/kg body wt by either oral or i.p. administration. Urine, feces and expired air were collected and at the end of 72 h the rats were sacrificed and tissues analyzed to determine 14C excretion and tissue distribution. After i.p. administration of methyl bromide, the dominant route of excretion was exhalation of 14CO2, with 46% of the dose exhaled as 14CO2. In contrast, urinary excretion of 14C was the major route of elimination (43% of the dose) when methyl bromide was given orally. Very little of the 14C appeared in the feces (less than 3% of the dose) regardless of route of administration. In rats with bile duct cannulations, 46% of an oral dose appeared in the bile over a 24-h period. Collection of bile significantly decreased the exhalation of 14CO2 and 14C excreted in urine compared to controls. At 72 h after oral or i.p. administration, 14-17% of the 14C remained in the rats, with liver and kidney being the major organs of retention. Results indicate that route of administration can affect the pathways for excretion. In addition, excretion of 14C in bile, coupled with the low levels of radioactivity found in the feces, indicates that reabsorption of biliary metabolites from the gut plays a significant role in the disposition of [14C] methyl bromide.

Administration, Oral↗

Some observations on contamination of animal cell cultures by the fungus Aspergillus fumigatus and suggested control measures.

The fungus Aspergillus fumigatus Fres. was found to be a most troublesome contaminant of animal cell cultures (hybridomas). Conditions in a carbon dioxide incubator (37 degrees C; about 100% relative humidity) are very favourable for its growth. Unlike bacteria it spreads by means of passively-released aerial spores and of filaments (mycelium) growing within and between wells of tissue culture vessels. It has the potential to cause mycotic infections and allergic reactions in laboratory workers. Various measures for controlling it in the tissue culture laboratory are briefly discussed, such as use of antifungal antibiotics, fumigation with formaldehyde and heat sterilisation of the incubator.

Amphotericin B↗

Mycorrhizae and their potential use in the agricultural and forestry industries.

Mycorrhizal fungi associated with plant roots increase the absorption of nutrients, particularly phosphorus, and thus enhance the growth of crop plants and trees. Vesicular-arbuscular mycorrhizae (VAM) occur in approximately 90% of all vascular plants including most of the important agricultural species, whereas ectomycorrhizae are found in most of the economically important tree species of the temperate regions of the world, and in some tropical trees. These symbiotic associations are, therefore, important in crop and biomass production. For this reason they are receiving considerable attention in agriculture and forestry. Currently, VAM are utilized in fumigated soils, greenhouse crops, and in the reclamation of disturbed sites. Ectomycorrhizae are employed in the establishment of trees in nurseries, in reforestation programs, and in the production of containerized seedlings. Production of VAM and ectomycorrhiza inoculum for large scale projects is now feasible but many basic questions related to persistence of these fungi in field situations, competition with other microorganisms, and particularly the most efficient fungi to use for particular hosts remain largely unanswered.

Journal Article↗

Evolution of enzymatic activity in the tautomerase superfamily: mechanistic and structural studies of the 1,3-dichloropropene catabolic enzymes.

The use of the soil fumigant Telone II, which contains a mixture of cis- and trans-1,3-dichloropropene, to control plant-parasitic nematodes is a common agricultural practice for maximizing yields of various crops. The effectiveness of Telone II is limited by the rapid turnover of the dichloropropenes in the soil due to the presence of bacterial catabolic pathways, which may be of recent origin. The characterization of three enzymes in these pathways, trans-3-chloroacrylic acid dehalogenase (CaaD), cis-3-chloroacrylic acid dehalogenase (cis-CaaD), and malonate semialdehyde decarboxylase (MSAD), has uncovered intriguing catalytic mechanisms as well as a fascinating evolutionary lineage for these proteins. Sequence comparisons and mutagenesis studies revealed that all three enzymes belong to the tautomerase superfamily. Tautomerase superfamily members with known structures are characterized by a beta-alpha-beta structural fold. Moreover, they have a conserved N-terminal proline, which plays an important catalytic role. Mechanistic, NMR, and pH rate studies of the two dehalogenases, coupled with a crystal structure of CaaD inactivated by 3-bromopropiolate, indicate that they use a general acid/base mechanism to catalyze the conversion of their respective isomer of 3-chloroacrylate to malonate semialdehyde. The reaction is initiated by the conjugate addition of water to the C-2, C-3 double bond and is followed by the loss of HCl. MSAD processes malonate semialdehyde to acetaldehyde, and is the first identified decarboxylase in the tautomerase superfamily. The catalytic mechanism is not well defined but the N-terminal proline plays a prominent role and may function as a general acid catalyst, similar to its role in CaaD and cis-CaaD. These are the first structural and mechanistic details for tautomerase superfamily members that catalyze either a hydration or a decarboxylation reaction, rather than a tautomerization reaction, in which Pro-1 serves as a general acid catalyst rather than as a general base catalyst. The available information on the 1,3-dichloropropene catabolic enzymes allows speculation on the possible evolutionary origins of their activities.

Allyl Compounds↗

Comparison of different microbial biomass and activity measurement methods in metal-contaminated soils.

The aims of this study were: (1) to compare different microbial methods of detecting the effects of heavy metals on the functioning of the soil ecosystem; and (2) to evaluate the effect of incubation on microbial biomass and microbial activity in soils that were not pre-incubated after sampling in order to determine their suitability for measuring the effects of heavy metals on the soil microbial ecosystem. The microbial biomass methods (included: biomass C, N and ninhydrin-N by fumigation-extraction (FE); substrate-induced respiration (SIR); soil ATP content and microbial activity as evolved CO2-C and arginine ammonification. All were tested in soils from the Woburn Market Garden Experiment. Due to past sludge application the soils contained, Zn, Cu or Ni at around current European Union upper limits and Cd at up to three times the limit. The amount of microbial biomass in metal-contaminated soils was about half of that found in soils from the experiment that received uncontaminated organic manure or inorganic fertilizer. The amount of biomass measured by FE and soil ATP content in incubated soils showed little change over 20 days incubation. However, SIR measurements were statistically affected over the first few days of incubation. The rates of arginine ammonification were higher in this order: farmyard manure (FYM)>inorganic fertilizer>sewage-sludge throughout the incubation. However, the evolved CO2-C rates were not significantly different among the treatments. Discriminant analysis confirmed smaller amounts of biomass in the metal-contaminated soils than in the other treatments. Linked properties, such as relationships between biomass and soil organic matter, or biomass-specific respiration rates, may provide "internal control" which may help overcome problems of establishing suitable control, or comparative measurements, when moving from experimental to natural environments.

Biodegradation, Environmental↗

Evolution of soil organic matter changes using pyrolysis and metabolic indices: a comparison between organic and mineral fertilization.

The aim of this study was to evaluate chemical and biochemical changes of organic matter in fertilized (ammonium nitrate) and amended (vermicompost and manure) soils using pyrolysis and metabolic indices. The metabolic potential [dehydrogenase (DH-ase)/water soluble organic carbon (WSOC)], the metabolic quotient (qCO2) and the microbial quotient (Cmic:Corg) were calculated as indices of soil organic matter evolution. Pyrolysis-gas chromatography (Py-GC) was used to study structural changes in the organic matter. Carbon forms and microbial biomass have been measured by dichromate oxidation and fumigation-extraction methods, respectively. Dehydrogenase activity has been tested using INT (p-Iodonitrotetrazolium violet) as substrate. The results showed that organic amendment increased soil microbial biomass and its activity which were strictly related to pyrolytic mineralization and humification indices (N/O, B/E3). Mineral fertilization caused a greater alteration of native soil organic matter than the organic amendments, in that a high release of WSOC and relatively large amounts of aliphatic pyrolytic products, were observed. Therefore, the pyrolysis and metabolic indices provided similar and complementary information on soil organic matter changes after mineral and organic fertilization.

Biomass↗

Effects of sulfonamide and tetracycline antibiotics on soil microbial activity and microbial biomass.

Increasingly often soil residual concentrations of pharmaceutical antibiotics are detected, while their ecotoxic relevance is scarcely known. Thus, dose related effects of two antibiotics, sulfapyridine and oxytetracycline, on microorganisms of two different topsoils were investigated. The fumigation-extracted microbial C (E(C)) and ergosterol were determined to indicate soil microbial and fungal biomass, respectively. Microbial activity was tested as basal respiration (BR), dehydrogenase activity (DHA), substrate-induced respiration (SIR), and Fe(III) reduction. The BR and DHA were uninfluenced even at antibiotic concentrations of 1000 microg g(-1). This revealed that an activation of microbial growth through nutrient substrate addition is required to test possible effects of the bacteriostatic antibiotics. In addition, the effects of both antibiotics were time dependent, showing that short-term tests were not suitable. Clear dose-response relations were determined with SIR when the short-term incubation of 4h was extended into the growth phase of the microorganisms (24 and 48 h). The Fe(III) reduction test, with a 7-d incubation, was also found to be suitable for toxicity testing of antibiotics in soils. Effective doses inhibiting the microbial activity by 10% (ED(10)) ranged from total antibiotic concentrations of 0.003-7.35 microg g(-1), depending on the antibiotic compound and its soil adsorption. Effective solution concentrations (EC(10)), calculated from distribution coefficients, ranged from 0.2 to 160 ng g(-1). The antibiotics significantly (p<0.05) reduced numbers of soil bacteria, resulting in dose related shifts in the fungal:bacterial ratio, which increased during 14 d, as determined from analysis of ergosterol and E(C). It was concluded that pharmaceutical antibiotics can exert a temporary selective pressure on soil microorganisms even at environmentally relevant concentrations.

Bacteria↗

Differential assimilation of nitrogen dioxide by 70 taxa of roadside trees at an urban pollution level.

In order to screen for the best species for mitigating nitrogen dioxide (NO2) by plants at urban levels, we investigated assimilation of nitrogen dioxide by 70 taxa of woody plants that are mostly utilized as roadside trees. They were fumigated with 15N-labeled NO2 at 0.1 microl l(-1) for 8h, and the amount of reduced nitrogen derived from NO2 (in mg Ng(-1) dry weight) in the leaves (designated NO2 assimilation capability hereafter) were determined. Data were analyzed in the comparison with the previously reported ones obtained at 4 microl l(-1) NO2. Among the 70 taxa, the value of NO2 assimilation capability differed by a factor of 122 between the highest (Prunus yedoensis; 0.061) and the lowest (Cryptomeria japonica; 0.0005). Based on the analysis of NO2 assimilation capability values at 0.1 and 4 micro l(-1) NO2, the 70 taxa of woody plants appeared to be classified into four types; those of high NO2 assimilation and high NO2 resistance, those of high NO2 assimilation but low NO2 resistance, those of low NO2 assimilation and low NO2 resistance, and those of low NO2 assimilation but high NO2 resistance. The first, second, third and fourth types include 13, 11, 35 and 11 taxa, respectively. The broad-leaf deciduous trees may have advantages of high biomass and fast growth as compared with woody plants of other habits. Thus, four broad-leaf deciduous species, Robinia pseudo-acacia, Sophora japonica, Populus nigra and Prunus lannesiana, were concluded here to be the best phytoremediators for the urban air.

Air Pollutants↗

An environmental fate, exposure and risk assessment of ethylene oxide from diffuse emissions.

Ethylene oxide (EO) is mainly used as a chemical intermediate and as a fumigant and sterilizing agent. Through its manufacturing and intended uses, EO may be released into the environment. Therefore, an assessment of the environmental significance of those potential emissions was conducted. Data were collected describing pertinent physical properties, degradation and other loss mechanisms that control the fate of EO in the environment. Available aquatic and terrestrial ecotoxicity data were assembled and used as the basis of calculating critical toxicity values to characterize hazard. Environmental compartment concentrations of EO were calculated using Level III fugacity-based modeling. Six scenarios were developed to account for different climatic conditions in various portions of the US. Finally, potential regional-scale risks to aquatic and terrestrial wildlife were determined. In the conceptual model that was developed in this assessment, EO diffuses into air, partitions between environmental compartments, is transported out of the different compartments via advection, and undergoes abiotic and biological degradation within each compartment. All known emissions within the continental USA were assumed to enter a modeled region roughly the size of the State of Ohio. Organisms (receptors) were assumed to dwell in both terrestrial and aquatic compartments. Receptors were assumed to include small mammals, soil invertebrates, water column (pelagic) organisms, and sediment benthos. The goal of this assessment was to characterize any potential adverse risks to terrestrial and aquatic wildlife populations. Hazard Quotients (HQ) were calculated by dividing predicted concentrations of EO in air, water, sediment, and soil by their critical toxicity values. Maximum calculated HQ values in air were 1.52x10(-7), in water were 1.17x10(-5), in sediment were 2.25x10(-4), and in soil were 1.37x10(-5). The results of this assessment suggest that EO as currently manufactured and used does not pose adverse risks to aquatic or terrestrial wildlife. In all cases, the HQ values were much less than the maximum desired HQ value of 1.0 (4,400-6,600,000 times), indicating that the potential for diffuse emissions of EO to pose adverse environmental risks is quite low.

Animals↗

Decomposition of heavy metal contaminated nettles (Urtica dioica L.) in soils subjected to heavy metal pollution by river sediments.

Two incubation experiments were conducted to evaluate differences in the microbial use of non-contaminated and heavy metal contaminated nettle (Urtica dioica L.) shoot residues in three soils subjected to heavy metal pollution (Zn, Pb, Cu, and Cd) by river sediments. The microbial use of shoot residues was monitored by changes in microbial biomass C, biomass N, biomass P, ergosterol, N mineralisation, CO(2) production and O(2) consumption rates. Microbial biomass C, N, and P were estimated by fumigation extraction. In the non-amended soils, the mean microbial biomass C to soil organic C ratio decreased from 2.3% in the low metal soil to 1.1% in the high metal soils. In the 42-d incubation experiment, the addition of 2% nettle residues resulted in markedly increased contents of microbial biomass P (+240%), biomass C (+270%), biomass N (+310%), and ergosterol (+360%). The relative increase in the four microbial properties was similar for the three soils and did not show any clear heavy metal effect. The contents of microbial biomass C, N and P and ergosterol contents declined approximately by 30% during the incubation as in the non-amended soils. The ratios microbial biomass C to N, microbial biomass C to P, and ergosterol to microbial biomass C remained constant at 5.2, 26, and 0.5%, respectively. In the 6-d incubation experiment, the respiratory quotient CO(2)/O(2) increased from 0.74 in the low metal soil to 1.58 in the high metal soil in the non-amended soils. In the treatments amended with 4% nettle residues, the respiratory quotient was constant at 1.13, without any effects of the three soils or the two nettle treatments. Contaminated nettle residues led generally to significantly lower N mineralisation, CO(2) production and O(2) consumption rates than non-contaminated nettle residues. However, the absolute differences were small.

Biomass↗

Genetic control of methyl halide production in Arabidopsis.

Methyl chloride (CH(3)Cl) and methyl bromide (CH(3)Br) are the primary carriers of natural chlorine and bromine, respectively, to the stratosphere, where they catalyze the destruction of ozone, whereas methyl iodide (CH(3)I) influences aerosol formation and ozone loss in the boundary layer. CH(3)Br is also an agricultural pesticide whose use is regulated by international agreement. Despite the economic and environmental importance of these methyl halides, their natural sources and biological production mechanisms are poorly understood. Besides CH(3)Br fumigation, important sources include oceans, biomass burning, tropical plants, salt marshes, and certain crops and fungi. Here, we demonstrate that the model plant Arabidopsis thaliana produces and emits methyl halides and that the enzyme primarily responsible for the production is encoded by the HARMLESS TO OZONE LAYER (HOL) gene. The encoded protein belongs to a group of methyltransferases capable of catalyzing the S-adenosyl-L-methionine (SAM)-dependent methylation of chloride (Cl(-)), bromide (Br(-)), and iodide (I(-)) to produce methyl halides. In mutant plants with the HOL gene disrupted, methyl halide production is largely eliminated. A phylogenetic analysis with the HOL gene suggests that the ability to produce methyl halides is widespread among vascular plants. This approach provides a genetic basis for understanding and predicting patterns of methyl halide production by plants.

Amino Acid Sequence↗

A chamberless field exposure system for ozone enrichment of short vegetation.

Only few studies have been conducted as yet which focus on the effects of rising tropospheric ozone levels on semi-natural vegetation under free-air conditions. A new technical approach was used to examine the response of calcareous grassland to ozone employing a chamberless fumigation system. Four different ozone regimes were applied (1-, 1.33-, 1.66- and 2-fold ambient air levels) with five replicates each. Ozone enrichment was carried out on circular plots of 2 m in diameter by a computer controlled exposure system. Transparent windscreens encircling each plot accelerated the mixing of ambient air and ozone released. Thus, the use of blowers could be avoided. The exposure system presented here is regarded as an appropriate technique for free-air trace gas enrichment on short vegetation avoiding microclimatic alterations known to affect plant growth and pollutant uptake. Furthermore, the chosen technical set-up was rather cost-effective. Hence, it enabled the establishment of a larger number of replications providing the basis for results of higher statistical power.

Air Pollutants↗

Ozone exposure induces the activation of leaf senescence-related processes and morphological and growth changes in seedlings of Mediterranean tree species.

Four Mediterranean tree taxa, Quercus ilex subsp. ilex, Quercus ilex subsp. ballota, Olea europaea cv. vulgaris and Ceratonia siliqua, were exposed to different ozone (O(3)) concentrations in open top chambers (OTCs) during 2 years. Three treatments were applied: charcoal-filtered air (CF), non-filtered air (NF) and non-filtered air plus 40 ppb(v) of O(3) (NF +). The photochemical maximal efficiency, Fv/Fm, decreased in NF + plants during the second year of exposure, especially during the most stressful Mediterranean seasons (winter and summer). An increase of delta(13)C was found in three of the four studied species during the first year of exposure. This finding was only maintained in C. siliqua during the second year. Decreases in the chlorophyll content were detected during the first year of fumigations in all the species studied, but not during the second year. The NF + treatment induced changes in foliar anatomical characteristics, especially in leaf mass per area (LMA) and spongy parenchyma thickness, which increased in some species. A reduction in N content and an increase in delta(15)N were found in all species during the second year when exposed in the NF + OTCs, suggesting a change in their retranslocation pattern linked to an acceleration of leaf senescence, as also indicated by the above mentioned biochemical and anatomical foliar changes. The two Q. ilex subspecies were the most sensitive species since the changes in N concentration, delta(15)N, chlorophyll, leaf area, LMA and biomass occurred at ambient O(3) concentrations. However, C. siliqua was the most responsive species (29% biomass reduction) when exposed to the NF + treatment, followed by the two Q. ilex subspecies (14-20%) and O. europaea (no significant reduction). Ozone resistance of the latter species was linked to some plant traits such as chlorophyll concentrations, or spongy parenchyma thickness.

Biomass↗

Ozone-induced changes in the chloroplast structure of conifer needles, and their use in ozone diagnostics.

Ozone induces characteristic symptoms in the chloroplasts of the needles of several coniferous species. Chloroplasts are (1) reduced in size and (2) the stroma is electron dense. Moreover, (3) these chloroplast alterations are more pronounced in the outer mesophyll cell layers and in the upper side of the needle compared to the inner layers and lower side. The syndrome, including the three symptoms (1)-(3), is found in the green needles of Scots pine and Norway spruce not only in the experimental fumigations, but also in mature trees in the field, and has potential for diagnosis of ozone stress. For sound ozone diagnostics all three symptoms must be present in the samples studied. The symptoms in relation to needle anatomy and physiology is discussed, and recommendations for sampling and analysis are given.

Acclimatization↗

Impact of short-term and long-term elevated CO2 on emission of carbonyls from adult Quercus petraea and Carpinus betulus trees.

The study aimed to elucidate the effect of elevated CO2 in short- and long-term experiments on the emission potential of carbonyls from mature oak (Quercus petraea) and hornbeam (Carpinus betulus). The trees were investigated using a canopy crane established in a mixed forest in Switzerland. Short-term CO2 experiments were performed on single leaves with a gas exchange measuring unit by increasing CO2 in the enclosure before and during the measurements. Long-term CO2 fumigation was performed for one vegetation period by a webFACE design. Short-term as well as long-term exposure of leaves resulted in no significant changes in carbonyl exchange in neither Q. petraea nor C. betulus. Carbonyl emission was not affected by stomatal closure under elevated CO2 levels. In contrast to the emission of isoprenoids from vegetation which is thought to be reduced at elevated atmospheric CO2, the exchange of carbonyls seems to be generally unaffected under these conditions.

Acetaldehyde↗