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Windy A Boyd

Publications and source records attributed to Windy A Boyd.

4 recordsLinked to original sources

Using transgenic Caenorhabditis elegans in soil toxicity testing.

Soil bioassays are important tools for evaluating toxicological effects within the terrestrial environment. The American Society for Testing and Materials E2172-01 Standard Guide outlines a method for conducting laboratory soil toxicity tests using the nematode Caenorhabditis elegans. This method is an efficient tool for extracting C. elegans from soil samples and can be carried out after a 24-h exposure period using relatively small amounts of soil. Drawbacks of this method include problems with (1) recovery of nematodes from soils containing a high percentage of organic matter, and (2) distinguishing indigenous nematode species from nematodes added for the laboratory test. Due in part to these issues, C. elegans has not been extensively accepted for use in soil testing. To address these concerns and improve upon the American Society for Testing and Materials method, this project focused on using transgenic strains of C. elegans carrying a GFP-expressing element. Lethality and behavior tests revealed that the transgenic nematodes respond similarly to the wild-type N2 strain, indicating that they can be used in the same manner in soil testing. The GFP marker is easily identifiable not only within soils containing a large amount of organic matter, but also in field-collected soils containing indigenous nematodes. These results support the use of transgenic GFP C. elegans in soil bioassays as a tool to further the reliability of laboratory toxicity tests.

Aluminum Silicates↗

The effects of metals and food availability on the behavior of Caenorhabditis elegans.

Caenorhabditis elegans, a nonparasitic soil nematode, was used to assess the combined effects of metal exposures and food availability on behavior. Movement was monitored using a computer tracking system after exposures to Cu, Pb, or Cd while feeding was measured as a change in optical density (deltaOD) of bacteria suspensions over the exposure period. After 24-h exposures at high and low bacteria concentrations, movement was decreased in a concentration-dependent fashion by Pb and Cd but feeding reductions were not directly proportional to exposure concentrations. Copper exposure induced concentration-dependent declines in feeding and movement regardless of bacteria concentration. The impact of 24-h metal exposures was apparently reduced by increasing food availability. Therefore, exposures were shortened to 4 h in an attempt to minimize starvation effects on movement. Although nematodes were immobilized following 24 h of food depravation, worms deprived of food during the 4-h exposure continued to feed and move after exposure. A bead-ingestion assay after 4-h exposures was also used as an additional means of assessing the effects of metals on feeding behavior. Ingestion was significantly reduced by all concentrations of metals tested, indicating its sensitivity as a sublethal assay. Feeding (deltaOD) during exposures exhibited similar trends as ingestion but was slightly less sensitive, while movement was the least sensitive assay of 4-h metal exposures to C. elegans. Assessment of multiple sublethal endpoints allowed for the determination of the separate and interactive effects of metals and food availability on C. elegans behavior.

Animals↗

Comparison of the sensitivity of three nematode species to copper and their utility in aquatic and soil toxicity tests.

Nematodes are useful organisms for aquatic and soil toxicity testing because of their abundance and diversity as well as their ease of culturing and maintenance in the laboratory. The nematode Caenorhabditis elegans has been used extensively in toxicity testing, but its sensitivity to metal exposures in relation to other nematodes remains unclear. In this study, we compare the sensitivity and ease of use of two other rhabditid nematodes, Panagrellus redivivus and Pristionchus pacificus, to C. elegans. Toxicity endpoints were chosen to investigate the effects of Cu on the survival of these nematodes after soil exposures and on the survival, reproduction, movement, and feeding behavior of nematodes after exposures in aquatic medium. In all lethality testing, P. pacificus was the most sensitive, C. elegans exhibited intermediate sensitivity, and P. redivivus was the least sensitive. Reproduction and movement of C. elegans and reproduction of P. pacificus were decreased 50% by similar concentrations of Cu (EC50s approximately 2 mg/L), but P. pacificus movement was less sensitive to Cu exposures (EC50 = 8 mg/L). Although all nematodes may be useful in lethality assays, using P. redivivus in toxicity tests is complicated by the presence of two sexes and difficulties in obtaining age-synchronized cultures. Pristionchus pacificus is an ideal acute toxicity-testing organism because of its sensitivity and ease of culturing. However, C. elegans appears to be more sensitive and therefore most useful in behavioral assays. Future studies of the relative sensitivities of nematodes in toxicity testing should continue to investigate additional toxicants, nematode species, and quantifications of sublethal effects after soil exposures.

Animals↗

Availability of metals to the nematode Caenorhabditis elegans: toxicity based on total concentrations in soil and extracted fractions.

Current regulation of metals in soils is based on total metal concentrations rather than on actual exposure concentrations. Considering the extreme variation in soil physicochemical properties, total concentrations are not reflective of the availability and resultant toxicity of metals in different soils. In this study, the availability of Cd, Cu, Ni, Pb, and Zn to the free-living soil nematode Caenorhabditis elegans was assessed after 24-h exposures in three soils using a sequential soil extraction procedure. Albany soil, sampled from southern Georgia, USA, is characterized by a high sand content, whereas Cecil soil from the Piedmont region of Georgia contains higher fractions of clay and organic matter. The final soil was an American Society for Testing and Materials (ASTM) artificial medium composed of peat, kaolin clay, sand, and calcium carbonate. Based on their composition, ASTM medium would sorb metals most strongly and Albany soil the least strongly. In fact, 24-h lethal concentrations to 50% (LC50s) of nematodes for the five metals as determined by the total metal concentration followed this trend. In addition, water-extractable metals were lowest in ASTM medium and highest in Albany soil when spiked at the same concentrations. Our data show the need to consider soil type when performing toxicological tests and establishing site-specific allowable metal concentrations in soil.

Absorption↗