Sediment Bioaccumulation Test with Lumbriculus variegatus: Effects of Organism Loading.

At contaminated sediment sites, the bioavailability of contaminants in sediments is assessed using sediment-bioaccumulation tests with Lumbriculus variegates (Lv). The testing protocols recommend that ratio of total organic carbon (TOC) in sediment to L. variegatus (dry weight) (TOC/Lv) should be no less than 50:1. Occasionally, this recommendation is not followed, especially with sediments having low TOC, e.g., <1 %. This study evaluated the impacts and resulting biases in the testing results when the recommendation of "no less than 50:1" is not followed. In the study, seven sediments were tested with a series of TOC/Lv ratios that spanned the recommendation. With increasing loading of organisms, growth of the organisms decreased in six of the seven sediments tested. Residues of polychlorinated biphenyls (PCBs) in the L. variegatus were measured in six of the seven sediments tested, and differences in PCB residues among loading ratios across all sediments were small, i.e., ±50 %, from those measured at the minimum recommended ratio of 50:1 TOC/Lv. In all sediment, PCB residues increased with increasing loading of the organisms for the mono-, di-, and tri-chloro-PCBs. For tetra-chloro and heavier PCBs, residues increased with increasing loading of organisms for only two of the six sediments. PCB residues were not significantly different between TOC/Lv loadings of 50:1 and mid-20:1 ratios indicating that equivalent results can be obtained with TOC/Lv ratios into the mid-20:1 ratios. Overall, the testing results suggest that when testing recommendation of 50:1 TOC/Lv is not followed, potential biases in the biota-sediment accumulations factors from the sediment-bioaccumulation test will be small.

Sediment bioaccumulation test with Lumbriculus variegatus: effects of feeding.

Standard sediment-bioaccumulation test methods specify that Lumbriculus variegatus should not be fed during the 28-day exposure. This lack of feeding can lead to decreases in L. variegatus weight and lipid content during the 28-day exposure period. Differences in intrinsic nutritional content of sediments could lead to additional variability in organism performance and/or contaminant uptake. To evaluate the potential benefits of feeding, sediment-bioaccumulation tests were performed comparing treatments with and without supplemental feeding with tropical fish food and also comparing performance food introduced as blended slurry versus fine flakes. The ration of food provided had to be limited to 6 mg/300-mL beaker with 250 mg of L. variegatus (ww) receiving three feedings per week to maintain acceptable dissolved oxygen (DO) in the test chambers. Relative weight change during exposure varied across sediments in the absence of food from very little change to as much as a 40 % decrease from starting weight. Feeding slurry and flake foods increased the total weight of recovered organisms by 32 and 48 %, respectively, but they did not decrease variability in weight changes across sediments. Lipid contents of the organisms decreased similarly across all feeding treatments during the test. At test termination, lipid contents of L. variegatus across unfed, slurry-fed, and flake-fed treatments were not significantly different per Tukey's honest significant difference test with 95 % family-wise confidence. Feeding resulted in polychlorinated biphenyl residues in L. variegatus being generally slightly less (median 78 %) and slightly greater (median 135 %) than the unfed treatments with slurry and flake formulated foods, respectively.

Evaluation of micro-colorimetric lipid determination method with samples prepared using sonication and accelerated solvent extraction methods.

Two common laboratory extraction techniques were evaluated for routine use with the micro-colorimetric lipid determination method developed by Van Handel (1985) [2] and recently validated for small samples by Inouye and Lotufo (2006) [1]. With the accelerated solvent extraction method using chloroform:methanol solvent and the colorimetric lipid determination method, 28 of 30 samples had significant proportional bias (α=1%, determined using standard additions) and 1 of 30 samples had significant constant bias (α=1%, determined using Youden Blank measurements). With sonic extraction, 0 of 6 samples had significant proportional bias (α=1%) and 1 of 6 samples had significant constant bias (α=1%). These demonstrate that the accelerated solvent extraction method with chloroform:methanol solvent system creates an interference with the colorimetric assay method, and without accounting for the bias in the analysis, inaccurate measurements would be obtained.