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Estimation of diversity and community structure through restriction fragment length polymorphism distribution analysis of bacterial 16S rRNA genes from a microbial mat at an active, hydrothermal vent system, Loihi Seamount, Hawaii.

PCR was used to amplify (eu)bacterial small-subunit (16S) rRNA genes from total-community genomic DNA. The source of total-community genomic DNA used for this culture-independent analysis was the microbial mats from a deep-sea, hydrothermal vent system, Pele's Vents, located at Loihi Seamount, Hawaii. Oligonucleotides complementary to conserved regions in the 16S rRNA-encoding DNA (rDNA) of bacteria were used to direct the synthesis of PCR products, which were then subcloned by blunt-end ligation into phagemid vector pBluescript II. Restriction fragment length polymorphism patterns, created by using tandem tetrameric restriction endonucleases, revealed the presence of 12 groups of 16S rRNA genes representing discrete operational taxonomic units (OTUs). The rank order abundance of these putative OTUs was measured, and the two most abundant OTUs accounted for 72.9% of all of the 16S rDNA clones. Among the remaining 27.1% of the 16S rDNA clones, none of the 10 OTUs was represented by more than three individual clones. The cumulative OTU distribution for 48 bacterial 16S rDNA clones demonstrated that the majority of taxa represented in the clone library were detected, a result which we assume to be an estimate of the diversity of bacteria in the native hydrothermal vent habitat. 16S rDNA fingerprinting of individual clones belonging to particular OTUs by using an oligonucleotide probe that binds to a universally conserved region of the 16S rDNA fragments was conducted to confirm OTU specificity and 16S rDNA identity.

Bacteria↗

Steady-state and transient-state performance of a biotrickling filter treating chlorobenzene-containing waste gas.

Biotrickling filter (BTF) technology was applied for the treatment of waste gas containing a mixture of chlorobenzene and 1,2-dichlorobenzene. An adapted microbial community was immobilised on a structured packing material. The strategy followed was to reach high removal efficiencies at initially low mass loading rates followed by an increase of the latter. This procedure was successful and resulted in a short start-up period of only 2 weeks. A 3-month operation under steady-state conditions showed good performance, with >95% removal efficiency at a mass loading rate of 1,800 g m(-3) day(-1). Dimensionless concentration profiles showed that the chlorobenzenes were simultaneously degraded. Low dissolved organic carbon of 15 mg l(-1) and stoichiometric chloride concentrations in the trickling liquid indicated complete mineralisation of the pollutant. Transient-state experiments with five times higher mass loading rates caused a decrease in the removal efficiency that recovered rapidly once the mass loading rate returned to its original steady-state level. A progressive increase of the mass loading rate in a long-term performance experiment showed that the removal efficiency could be kept stable between 95 and 99% at loads of up to 5,200 g m(-3) day(-1) over several days. Above this mass loading rate, the elimination capacity did not increase any further. These results demonstrated that with a well-adapted inoculum and optimal operation parameters, a BTF system with excellent performance and stability that efficiently removes a mixture of chlorobenzene vapours from air can be obtained.

Air Pollutants↗

Nitrogen availability to Pseudomonas fluorescens DF57 is limited during decomposition of barley straw in bulk soil and in the barley rhizosphere.

The availability of nitrogen to Pseudomonas fluorescens DF57 during straw degradation in bulk soil and in barley rhizosphere was studied by introducing a bioluminescent reporter strain (DF57-N3), responding to nitrogen limitation, to model systems of varying complexity. DF57-N3 was apparently not nitrogen limited in the natural and sterilized bulk soil used for these experiments. The soil was subsequently amended with barley straw, representing a plant residue with a high carbon-to-nitrogen ratio (between 60 and 100). In these systems the DF57-N3 population gradually developed a nitrogen limitation response during the first week of straw decomposition, but exclusively in the presence of the indigenous microbial population. This probably reflects the restricted ability of DF57 to degrade plant polymers by hydrolytic enzymes. The impact of the indigenous population on nitrogen availability to DF57-N3 was mimicked by the cellulolytic organism Trichoderma harzianum Rifai strain T3 when coinoculated with DF57-N3 in sterilized, straw-amended soil. Limitation occurred concomitantly with fungal cellulase production, pointing to the significance of hydrolytic activity for the mobilization of straw carbon sources, thereby increasing the nitrogen demand. Enhanced survival of DF57-N3 in natural soil after straw amendment further indicated that DF57 was cross-fed with carbon/energy sources. The natural barley rhizosphere was experienced by DF57-N3 as an environment with restricted nitrogen availability regardless of straw amendment. In the rhizosphere of plants grown in sterilized soil, nitrogen limitation was less severe, pointing to competition with indigenous microorganisms as an important determinant of the nitrogen status for DF57-N3 in this environment. Hence, these studies have demonstrated that nitrogen availability and gene expression in Pseudomonas is intimately linked to the structure and function of the microbial community. Further, it was demonstrated that the activities of cellulolytic microorganisms may affect the availability of energy and specific nutrients to a group of organisms deficient in hydrolytic enzyme activities.

Biodegradation, Environmental↗

Fatty acid-oxidizing consortia along a nutrient gradient in the Florida Everglades.

The Florida Everglades is one of the largest freshwater marshes in North America and has been subject to eutrophication for decades. A gradient in P concentrations extends for several kilometers into the interior of the northern regions of the marsh, and the structure and function of soil microbial communities vary along the gradient. In this study, stable isotope probing was employed to investigate the fate of carbon from the fermentation products propionate and butyrate in soils from three sites along the nutrient gradient. For propionate microcosms, 16S rRNA gene clone libraries from eutrophic and transition sites were dominated by sequences related to previously described propionate oxidizers, such as Pelotomaculum spp. and Syntrophobacter spp. Significant representation was also observed for sequences related to Smithella propionica, which dismutates propionate to butyrate. Sequences of dominant phylotypes from oligotrophic samples did not cluster with known syntrophs but with sulfate-reducing prokaryotes (SRP) and Pelobacter spp. In butyrate microcosms, sequences clustering with Syntrophospora spp. and Syntrophomonas spp. dominated eutrophic microcosms, and sequences related to Pelospora dominated the transition microcosm. Sequences related to Pelospora spp. and SRP dominated clone libraries from oligotrophic microcosms. Sequences from diverse bacterial phyla and primary fermenters were also present in most libraries. Archaeal sequences from eutrophic microcosms included sequences characteristic of Methanomicrobiaceae, Methanospirillaceae, and Methanosaetaceae. Oligotrophic microcosms were dominated by acetotrophs, including sequences related to Methanosarcina, suggesting accumulation of acetate.

Bacteria↗

Influence of substrate composition and flow rate on growth of Azospirillum brasilense Cd in a co-culture with 3 sorghum rhizobacteria.

The ability of Azospirillum brasilense Cd to colonize the niche occupied by 3 bacterial strains previously isolated from sorghum rhizosphere was studied by means of the Biolog system. The isolates were identified by different methods as strains belonging to Pseudomonas putida, Stenotrophomonas maltophilia, and Klebsiella terrigena species. Several C sources, also chosen among the constituents of sorghum root exudates, were used to evaluate the metabolic profiles of Azospirillum and the sorghum rhizobacteria. Azospirillum brasilense Cd exploited the same class of C compounds as the sorghum rhizobacteria and overlapped in their niche requirements. Since structure and functioning of a microbial community are largely affected by the flow rate of nutrient supply, the competitive behavior of A. brasilense Cd was studied in a chemostat mixed culture under C-limited conditions using disodium succinate as C source. Only at high growth rates, i.e., when the C source was highly supplied, A. brasilense Cd appeared to be a good competitor and it became the dominant species, whereas at low growth rates, it was outnumbered by the other species. However, the coexistence of all the strains was always maintained, thus suggesting that interactions other than competition or a potential cross-feeding might occur within the mixed culture.

Amino Acids↗

Oxygen respiration by desulfovibrio species.

Throughout the first 90 years after their discovery, sulfate-reducing bacteria were thought to be strict anaerobes. During the last 15 years, however, it has turned out that they have manifold properties that enable them to cope with oxygen. Sulfate-reducing bacteria not only survive oxygen exposure for at least days, but many of them even reduce oxygen to water. This process can be a true respiration process when it is coupled to energy conservation. Various oxygen-reducing systems are present in Desulfovibrio species. In Desulfovibrio vulgaris and Desulfovibrio desulfuricans, oxygen reduction was coupled to proton translocation and ATP conservation. In these species, the periplasmic fraction, which contains hydrogenase and cytochrome c3, was found to catalyze oxygen reduction with high rates. In Desulfovibrio gigas, a cytoplasmic rubredoxin oxidase was identified as an oxygen-reducing terminal oxidase. Generally, the same substrates as with sulfate are oxidized with oxygen. As additional electron donors, reduced sulfur compounds can be oxidized to sulfate. Sulfate-reducing bacteria are thus able to catalyze all reactions of a complete sulfur cycle. Despite a high respiration rate and energy coupling, aerobic growth of pure cultures is poor or absent. Instead, the respiration capacity appears to have a protective function. High numbers of sulfate-reducing bacteria are present in the oxic zones and near the oxic-anoxic boundaries of sediments and in stratified water bodies, microbial mats and termite guts. Community structure analyses and microbiological studies have shown that the populations in those zones are especially adapted to oxygen. How dissimilatory sulfate reduction can occur in the presence of oxygen is still enigmatic, because in pure culture oxygen blocks sulfate reduction. Behavioral responses to oxygen include aggregation, migration to anoxic zones, and aerotaxis. The latter leads to band formation in oxygen-containing zones at concentrations of </=20% air saturation.

Adaptation, Biological↗

Toxicity of pesticides to aquatic microorganisms: a review.

Microorganisms contribute significantly to primary production, nutrient cycling, and decomposition in estuarine eco-systems; therefore, detrimental effects of pesticides on microbial species may have subsequent impacts on higher trophic levels. Pesticides may affect estuarine microorganisms via spills, runoff, and drift. Both the structure and the function of microbial communities may be impaired by pesticide toxicity. Pesticides may also be metabolized or bioaccumulated by microorganisms. Mechanisms of toxicity vary, depending on the type of pesticide and the microbial species exposed. Herbicides are generally most toxic to phototrophic microorganisms, exhibiting toxicity by disrupting photosynthesis. Atrazine is the most widely used and most extensively studied herbicide. Toxic effects of organophosphate and organochlorine insecticides on microbial species have also been demonstrated, although their mechanisms of toxicity in such nontarget species remain unclear. There is a great deal of variability in the toxicity of even a single pesticide among microbial species. When attempting to predict the toxicity of pesticides in estuarine ecosystems, effects of pesticide mixtures and interactions with nutrients should be considered. The toxicity of pesticides to aquatic microorganisms, especially bacteria and protozoa, is an area of research requiring further study.

Animals↗

[Biogeochemical processes in the algal-bacterial mats of the Urinskii alkaline hot spring].

The structure and production characteristics of microbial communities from the Urinskii alkaline hot spring (Buryat Republic, Russia) have been investigated. A distinctive characteristic of this hot spring is the lack of sulfide in the issuing water. The water temperature near the spring vents ranged from 69 to 38.5 degrees C and pH values ranged from 8.8 to 9.2. The total mineralization of water was less than 0.1 g/liter. Temperature has a profound effect on the species composition and biogeochemical processes occurring in the algal-bacterial mats of the Urinskii hot spring. The maximum diversity of the phototrophic community was observed at the temperatures 40 and 46 degrees C. A total of 12 species of cyanobacteria, 4 species of diatoms, and one species of thermophilic anoxygenic phototrophic bacteria, Chloroflexus aurantiacus, have been isolated from mat samples. At temperatures above 40 degrees C, the filamentous cyanobacterium Phormidium laminosum was predominant; its cell number and biomass concentration were 95.1 and 63.9%, respectively. At lower temperatures, the biomass concentrations of the cyanobacterium Oscillatoria limosa and diatoms increased (50.2 and 36.4%, respectively). The cyanobacterium Mastigocladus laminosus, which is normally found in neutral or slightly acidic hydrothermal systems, was detected in microbial communities. As the diatom concentration increases, so does the dry matter concentration in mats, while the content of organic matter decreases. The concentrations of proteins and carbohydrates reached their maximum levels at 45-50 degrees C. The maximum average rate of oxygenic photosynthesis (2.1 g C/m2 day), chlorophyll a content (343.4 mg/m2), and cell number of phototrophic microorganisms were observed at temperatures from 45 to 50 degrees C. The peak mass of bacterial mats (56.75 g/m2) occurred at a temperature of 65-60 degrees C. The maximum biomass concentration of phototrophs (414.63 x 10(-6) g/ml) and the peak rate of anoxygenic photosynthesis [0.42 g C/(m2 day)] were observed at a temperature of 35-40 degrees C.

Algal Proteins↗

Problems and potential for in situ treatment of environmental pollutants by engineered microorganisms.

Molecular microbial ecology could provide tools for studying the structure and function of biodegradative microbial communities useful for in situ removal of environmental pollutants. Detection and monitoring of released organisms in the environment is required and DNA:DNA colony hybridization seems to be one of the highly sensitive and accurate technologies available for achieving this goal. A realistic and pragmatic view is required for analysis of the risks of environmental release of genetically-modified or engineered organisms.

Bacteria↗

[The effectiveness of using biohumus for growing plants under autonomous conditions].

There has been investigated the possibility of prolonging the useful time of the substrate on the basis of natural salt-loaded zeolite "Balkanin" used in space greenhouses to grow plants through introduction of an organic addition, i.e. biohumus, instead of regeneration. Biohumus has been applied into the substrate in amounts of 5.0-15.0 wt%. There has been grown tropical perennial plant Basella rubra L. used for food as a salad. It is found that the periodic introduction of biohumus in substrate enables a 4-19 time increase of its useful life without regeneration within more than 4 years and assures high and stable crop yield with high quality biomass. As compared to the control, the nitrates content of the plant biomass was, on the average, 3 times lower and decreased to the point of their total absence at the end of experiment. The results obtained allow us to recommend the above-mentioned technology of plant growing in space greenhouses both when included into closed environmental systems and in independent plant growth facilities aboard spacecraft. During space flights exceeding 2 years in length, additional supplies of biohumus will be required. This will be enough to stabilize the structure and the size of microbial community in substrate and provide a high crop yield.

Manure↗

Metaproteomic analysis of Chesapeake Bay microbial communities.

BACKGROUND: Natural microbial communities are extremely complex and dynamic systems in terms of their population structure and functions. However, little is known about the in situ functions of the microbial communities. RESULTS: This study describes the application of proteomic approaches (metaproteomics) to observe expressed protein profiles of natural microbial communities (metaproteomes). The technique was validated using a constructed community and subsequently used to analyze Chesapeake Bay microbial community (0.2 to 3.0 microm) metaproteomes. Chesapeake Bay metaproteomes contained proteins from pI 4-8 with apparent molecular masses between 10-80 kDa. Replicated middle Bay metaproteomes shared approximately 92% of all detected spots, but only shared 30% and 70% of common protein spots with upper and lower Bay metaproteomes. MALDI-TOF analysis of highly expressed proteins produced no significant matches to known proteins. Three Chesapeake Bay proteins were tentatively identified by LC-MS/MS sequencing coupled with MS-BLAST searching. The proteins identified were of marine microbial origin and correlated with abundant Chesapeake Bay microbial lineages, Bacteroides and alpha-proteobacteria. CONCLUSION: Our results represent the first metaproteomic study of aquatic microbial assemblages and demonstrate the potential of metaproteomic approaches to link metagenomic data, taxonomic diversity, functional diversity and biological processes in natural environments.

Journal Article↗

Experimental manipulations of microbial food web interactions in a humic lake: shifting biological drivers of bacterial community structure.

A previous multiyear study observed correlations between bacterioplankton community composition (BCC) and abundance and the dynamics of phytoplankton populations and bacterivorous grazers in a humic lake. These observations generated hypotheses about the importance of trophic interactions (both top-down and bottom-up) for structuring bacterial communities in this lake, which were tested using two multifactorial food web manipulation experiments that separately manipulated the intensity of grazing and the composition of the phytoplankton community. Our results, combined with field observations, suggest that a hierarchy of drivers structures bacterial communities in this lake. While other studies have noted links between aggregate measures of phytoplankton and bacterioplankton communities, we demonstrate here correlations between succession of phytoplankton assemblages and BCC as assessed by automated ribosomal intergenic spacer analysis (ARISA). We used a novel approach linking community ARISA data to phylogenetic assignments from sequence analysis of 16S rRNA gene clone libraries to examine the responses of specific bacterial phylotypes to the experimental manipulations. The synchronous dynamics of these populations suggests that primary producers may mediate BCC and diversity through labile organic matter production, which evolves in quality and quantity during phytoplankton succession. Superimposed on this resource-mediated control of BCC are brief periods of intense bacterivory that impact bacterial abundance and composition.

Animals↗

A geostatistical analysis of small-scale spatial variability in bacterial abundance and community structure in salt marsh creek bank sediments.

Small-scale variations in bacterial abundance and community structure were examined in salt marsh sediments from Virginia's eastern shore. Samples were collected at 5 cm intervals (horizontally) along a 50 cm elevation gradient, over a 215 cm horizontal transect. For each sample, bacterial abundance was determined using acridine orange direct counts and community structure was analyzed using randomly amplified polymorphic DNA fingerprinting of whole-community DNA extracts. A geostatistical analysis was used to determine the degree of spatial autocorrelation among the samples, for each variable and each direction (horizontal and vertical). The proportion of variance in bacterial abundance that could be accounted for by the spatial model was quite high (vertical: 60%, horizontal: 73%); significant autocorrelation was found among samples separated by 25 cm in the vertical direction and up to 115 cm horizontally. In contrast, most of the variability in community structure was not accounted for by simply considering the spatial separation of samples (vertical: 11%, horizontal: 22%), and must reflect variability from other parameters (e.g., variation at other spatial scales, experimental error, or environmental heterogeneity). Microbial community patch size based upon overall similarity in community structure varied between 17 cm (vertical) and 35 cm (horizontal). Overall, variability due to horizontal position (distance from the creek bank) was much smaller than that due to vertical position (elevation) for both community properties assayed. This suggests that processes more correlated with elevation (e.g., drainage and redox potential) vary at a smaller scale (therefore producing smaller patch sizes) than processes controlled by distance from the creek bank.

Bacteria↗

Dental plaque: biological significance of a biofilm and community life-style.

BACKGROUND: Most microorganisms in nature attach to surfaces and form matrix-embedded biofilms. Biofilms are highly structured and spatially organized, and are often composed of consortia of interacting microorganisms, termed microbial communities, the properties of which are more than the sum of the component species. Microbial gene expression alters markedly in biofilms; organisms communicate by gene transfer and by secretion of diffusible signalling molecules. Cells in biofilms are less susceptible to antimicrobial agents. AIM AND MATERIALS & METHODS: To comprehensively review the literature to determine whether dental plaque displays properties consistent with those of a typical biofilm and microbial community. RESULTS: Novel microscopic and molecular techniques have demonstrated that plaque has a structured architecture with an extracellular matrix, and a diverse composition (around 50% of cells are unculturable). The constituent species communicate by gene transfer, by secreted peptides (gram-positive bacteria) and autoinducer-2 (gram-positive and gram-negative bacteria). These organisms are functionally organized for increased metabolic efficiency, greater resistance to stress and for enhanced virulence. Plaque formation has direct and indirect effects on gene expression. CONCLUSION: Dental plaque displays properties that are typical of biofilms and microbial communities in general, a clinical consequence of which is a reduced susceptibility to antimicrobial agents as well as pathogenic synergism.

Biofilms↗

Molecular microbial biodiversity assessment: a biological indicator of soil health.

The soil performs a variety of key functions: (i) provides the food, fuel, and fiber needs of the world's population, (ii) regulates the quality of the air and water, (iii) decomposes organic wastes, (iv) recycles nutrients, and (v) acts as a sink for pollutants (including global gases). Soil degradation is increasingly recognized as an urgent environmental issue and a crucial need exists for the capacity to evaluate soil health. After all, soil health is a key component in sustaining the different world's ecosystems and the myriad of natural and socioeconomic systems they support. In this respect, soil health has been defined as the "continued capacity of a specific kind of soil to function as a vital living system, within natural or managed ecosystem boundaries, to sustain plant and animal productivity, to maintain or enhance the quality of air and water environments, and to support human health and habitation". A biological indicator is an organism, a part of an organism, or a community of organisms, used to obtain information about environmental quality. The assessment of microbial biodiversity has the potential to provide useful insight into the health and functioning of soil. Our inability to culture most microorganisms that are present in soils has, until recently, impaired studying the relationships between the structure and the function of soil microbial communities. This shortcoming has been recently overcome by using several molecular techniques that allow the detection, enumeration, and characterization of soil microorganisms without cultivation.

Bacteria↗

Transgenic maize in the presence of ampicillin modifies the metabolic profile and microbial population structure of bovine rumen fluid in vitro.

Recently, transgenic crops have been considered as possible donors of transgenes that could be taken up by micro-organisms under appropriate conditions. In an in vitro rumen simulation system, effects of ampicillin on microbial communities growing either on rumen contents with transgenic maize carrying a gene that confers resistance to ampicillin or its isogenic counterpart as substrates were examined continuously over 13 d. Rate of production of SCFA was measured to determine functional changes in the rumen model and single-strand conformational polymorphism was used to detect alterations in structure of the microbial community. Rumen contents treated with ampicillin displayed a marked decrease in the rate of production of SCFA and diversity of the microbial community was reduced severely. In the presence of transgenic maize, however, the patterns of change of rumen micro-organisms and their metabolic profiles were different from that of rumen fluid incorporating maize bred conventionally. Recovery of propionate production was observed both in the rumen fluid fed transgenic and conventional maize after a delay of several days but recovery occurred earlier in fermenters fed transgenic maize. Alterations in the microbial population structures resulting from the ampicillin challenge were not reversed during the experimental run although there was evidence of adaptation of the microbial communities over time in the presence of the antibiotic, showing that populations with different microbial structures could resume a pre-challenge metabolic profile following the introduction of ampicillin, irrespective of the source of the plant material in the growth medium.

Ampicillin↗

Structural diversity of microorganisms in chemically perturbed soil assessed by molecular and cytochemical approaches.

Until recently, our understanding of microbial community development in soil ecosystems exposed to different inorganic and organic pollutants has been limited to culturable microorganisms because of the techniques available. The discovery that most soil microorganisms are non-culturable but potentially viable and metabolically active accelerated the application of different culture-independent methods for structural diversity assessments of the microbial community. This review examines the results of recent studies on the impact of heavy metals and organic pollutants on the diversity of the microflora obtained with methods based on analyses of signature biomarkers such as nucleic acids and fatty acids. The application of these techniques allowed researchers to pinpoint reduction of microbial diversity in contaminated soil, and significant shifts in the community structure, leading to the dominance of only a few populations (species) and the disappearance of others, some of which were never isolated by conventional methods (e.g. an increase in Acidobacterium or a decrease in terrestrial non-thermophilic Crenarchaeota). Although the new techniques are not free from limitations, they allow the monitoring of the virtual impact of stressors on soil microorganisms and the direction of resuscitation of the microbial community during natural or induced bioremediation, especially when using combined approaches.

Bacteria↗

The use of integrated soil microcosms to assess the impact of carbendazim on soil ecosystems.

Our investigation used carbendazim as a representative pesticide for testing an integrated soil microcosm (ISM) test protocol. Microcosms, set up in a greenhouse, consisted of cylinders made from high-density polyethylene (HDPE) pipe, 7.5 cm (i.d.) x 15 cm high. A fine nylon mesh was placed across the bottom of each microcosm for leachate collection. Field soil, (silty clay loam), collected from Flörsheim, Germany, was sieved through a 5 mm screen and mixed thoroughly. Earthworms, enchytraeids, and microarthropods were added to each microcosm. Each microcosm contained five wheat seedlings, and was maintained at a 12 h-12 h light-dark cycle. Artificial rainwater was used to water microcosms as required. Soil microcosms were treated with carbendazim at concentrations 1, 3, 9, 27, and 81 times higher than the predicted environmental concentration (PEC) of 0.76 mg a.i./kg soil dry weight. A water-only control treatment was also used. The key soil processes used as endpoints were microbial activity, nitrogen mineralization. soil enzymatic activity, ammonium and nitrate leaching, organic matter decomposition and biological feeding activity. Key structural parameters measured were microbial biomass, nematode communities, microarthropod populations and diversity, enchytraeid and earthworm populations and plant growth. Pesticide degradation, leaching and uptake into plants and earthworms were also assessed. Carbendazim had significant effects on several key soil processes including soil ammonium-N and nitrate-N concentrations and soil dehydrogenase activity. Wheat growth, nematode and earthworm populations, and invertebrate feeding activity were soil structural parameters affected significantly by carbendazim. Earthworm biomass was the most sensitive parameter measured with an EC50 of 1.9 mg a.i./kg soil dry weight 28 days after treatment. A comparison of these results with results from single-species tests, small microcosms, large terrestrial model ecosystems, and field tests indicated that the ISM protocol may adequately predict environmental effects.

Animals↗