Characterization of microbial communities from coastal waters using microarrays

Molecular methods, including DNA probes, were used to identify and enumerate pathogenic Vibrio species in the Chesapeake Bay; our data indicated that Vibrio vulnificus exhibits seasonal fluctuations in number. Our work included a characterization of total microbial communities from the Bay; development of microarrays that identify and quantify the diversity of those communities; and observation of temporal changes in those communities. To identify members of the microbial community, we amplified the 16S rDNA gene from community DNA isolated from a biofilm sample collected from the Chesapeake Bay in February, 2000. The resultant 75 sequences were 95% or more similar to 7 species including two recently described Shewanella species, baltica and frigidimarina, that have not been previously isolated from the Chesapeake. When the genera of bacteria from biofilm after culturing are compared to those detected by subcloning amplified 16S fragments from community DNA, the cultured sample exhibited a strong bias. In oysters collected in February, the most common bacteria were previously unknown. Based on our 16S findings, we are developing microarrays to detect these and other microbial species in these estuarine communities. The microarrays will detect each species using four distinct loci, with the multiple loci serving as an internal control. The accuracy of the microarray will be measured using sentinel species such as Aeromonas species, Escherichia coli, and Vibrio vulnificus. Using microarrays, it should be possible to determine the annual fluctuations of bacterial species (culturable and non-culturable, pathogenic and non-pathogenic). The data may be applied to understanding patterns of environmental change; assessing the health of the Bay; and evaluating the risk of human illness associated with exposure to and ingestion of water and shellfish.     

Remediation of acid mine drainage leachate by a physicochemical and biological treatment‐train approach

Biological and physicochemical approaches were utilized in a treatment train for acid mine dis charge (AMD) waters. Anaerobic bioreactors, chemical precipitation reactors, and biopolymer chelation reactors, operated in static, semicontinuous, and continuous flow modes, removed significant quantities of metals and sulfates associated with AMD water. Static tests indicated accept able copper removal via precipitation by generation of hydrogen sulfide in anaerobic reactors. However, low pH affected the biopolymer coating in the chelation reactor, resulting in loss of bed surface. Corrections of AMD to pH > 7 resulted in some metal precipitationprior to biopolymer treatment. A series of static semicontinuous tests at pH 5.0 provided improved metal and sulfate removal. Copper (Cu⁺) was reduced to trace concentrations, while manganese (Mn⁺), although reduced, proved to be the most recalcitrant of the metals. © 2006 Wiley Periodicals, Inc.     

Unreliability of co‐occurrence‐based sediment quality guidelines for contaminated sediment evaluations at Superfund/hazardous chemical sites

Many Superfund/hazardous chemical sites include waterbodies whose sediments contain hazardous chemicals. With the need to assess, rank, and remediate contaminated sediments at such sites, as well as in other waterways, regulators seek a simple, quantitative assessment approach that feeds easily into a decision‐making scheme. Numeric, co‐occurrence‐based “sediment quality guidelines” have emerged with the appearance of administrative simplicity. However, the very foundation of the co‐occurrence approach, based on the total concentrations of a chemical(s) in sediment, is technically invalid; its application relies on additional technically invalid presumptions. Use of technically invalid evaluation approaches renders any assessment of the significance of sediment contamination unreliable. This article reviews the technical roots and assumptions of the co‐occurrence‐based SQGs, the fundamental flaws in the rationale behind their development and application, and their misapplication for sediment quality evaluation. It also reviews concepts and approaches for the more reliable evaluation, ranking, and cleanup assessment of contaminated sediments at Superfund sites and elsewhere. © 2005 Wiley Periodicals, Inc.     

Effects of commercial diazinon and imidacloprid on microbial urease activity in soil and sod

Diazinon [O,O-diethyl O-2-isopropyl-6-methyl(pyrimidine-4-yl) phosphorothioate] and imidacloprid [1-(1-[6-chloro-3-pyridinyl]methyl)-N-nitro-2-imidazolidinimine] are applied to lawns for insect control simultaneously with nitrogenous fertilizers such as urea, but their potential effect on urease activity and nitrogen availability in turfgrass management has not been evaluated. Urease activity in enzyme assays, washed cell assays, and soil slurries was examined as a function of insecticide concentration. Intact cores from field sites were used to assess the effect of insecticide application on urease activity in creeping bentgrass (Agrostis palustris Huds.) and bluegrass (Poa pratensis L.) sod. Bacterial urease from Bacillus pasteurii and plant urease from jack bean [Canavalia ensiformis (L.) DC.] were unaffected by the insecticides. Both insecticides inhibited the growth of Proteus vulgaris, a urease-producing bacterium, but only diazinon significantly reduced urease activity in washed cells; neither insecticide inhibited urease activity in sonicated cells. Neither diazinon nor imidacloprid inhibited urease activity in Woolper soil (fine, mixed, mesic Typic Argiudoll) slurries, but diazinon slightly inhibited urease activity in Maury soil (fine, mixed, semiactive, mesic Typic Paleudalf) slurries. Imidacloprid had no effect on urease activity in creeping bentgrass or bluegrass sod at up to 10 times the commercial application rate. Diazinon briefly, but significantly, reduced urease activity in bluegrass sod. Co-application of imidacloprid and urea appears to be benign with respect to urease activity in soil and sod. Diazinon, in contrast, appears to have a significant, short-term, inhibitory effect on the microbial urease-producing community, but that effect depends on soil type.     

Implications of urine-to-feces ratio in the thermophilic anaerobic digestion of swine waste.

Thermophilic anaerobic digestion of swine manure represents a potential waste treatment technology to address environmental concerns, such as odor emissions and removal of pathogenic microorganisms. However, there are concerns relative to the stability of this process when swine manure is the sole substrate. In this study, the potential of biogas production from swine manure as the sole substrate under thermophilic (50 degrees C) conditions was investigated in the laboratory, to determine whether separation of urine and feces as part of the waste collection process would benefit anaerobic digestion. Effluent from a continuously stirred tank reactor was used as the inoculum for batch tests, in which the substrate contained three different concentrations of urine (urine-free, as-excreted urine-to-feces ratio and double the as-excreted urine-to-feces ratio). Inocula were acclimated to these same urine-to-feces ratios to determine methane production. Results show that both urine-free and as-excreted substrates were not inhibitory to anaerobic inocula. Anaerobic microorganisms can be readily acclimated to substrate with double the as-excreted urine concentration, which contained nitrogen concentrations up to 7.20 g/L. Cumulative methane production reached similar levels in the batch tests, regardless of the substrate urine concentration.