Carlton T. Phillips

Reproduction and survival of Eisenia fetida in a sandy loam soil amended with the nitro-heterocyclic explosives RDX and HMX

Munitions manufacturing, disposal, testing, training and other operations at military sites have produced elevated levels of explosives and related materials in soil. Insufficient data were available on the toxicity of the explosives, RDX and HMX to soil invertebrates, necessitating toxicity testing. The U.S. Environmental Protection Agency (USEPA) in conjunction with stakeholders, is developing Ecological Soil Screening Level (Eco-SSL) benchmarks for ecological risk assessment (ERA) of contaminants at Superfund sites to identify those contaminants in soil that warrant additional evaluation in a baseline ERA, and to eliminate those that do not. Eco-SSLs are developed from literature values whenever sufficient quantity and quality of data exist. Tests were conducted under conditions preferred for Eco-SSL benchmarks, using a sandy loam soil that supports relatively high bioavailability of test compounds. Earthworm (Eisenia fetida) toxicity was assessed using a standardized earthworm reproduction test in freshly amended soil and weathered/aged amended soil. RDX or HMX had no effect on adult survival. Cocoon production EC 2 0 values for RDX were 1.2 and 19 mg kg - 1 in freshly amended and weathered/aged soils, respectively. Juvenile production EC 2 0 values were 1.6 and 4.8 mg kg - 1 in freshly amended and weathered/aged soils, respectively. Cocoon production and juvenile production EC 2 0 values for HMX were 2.7 and 0.4 mg kg - 1 in freshly amended soil. Both cocoon production and juvenile production in weathered/aged HMX treated soils were not significantly different(p > 0.05) from control soils. Results of these toxicity studies will be submitted to the Eco-SSLTask Group and will be included in the Eco-SSL database for Eco-SSL derivation.

Toxicity benchmarks for antimony, barium, and beryllium determined using reproduction endpoints for Folsomia candida, Eisenia fetida, and Enchytraeus crypticus

The U.S. Environmental Protection Agency is developing ecological soil screening levels (Eco-SSLs) for the ecological risk assessment of contaminants at Superfund sites. The Eco-SSLs for several soil contaminants have been developed from toxicity benchmarks for soil invertebrates in the existing literature. Insufficient information to generate Eco-SSLs for Sb, Ba, and Be necessitated toxicity testing to fill the data gaps. We used standardized toxicity tests with the earthworm Eiseniafetida, enchytraeid Enchytraeus crypticus, and collembolan Folsomia candida in the present study. These tests were selected on the basis of their ability to measure chemical toxicity to ecologically relevant test species during chronic assays and their inclusion of at least one reproduction component among the measurement endpoints. Tests were conducted in Sassafras Sandy Loam soil, which supports relatively high bioavailability of metals. Weathering and aging procedures for metals in amended soil were incorporated into these studies to better reflect exposure conditions in the field. The relative toxicity of metals to the soil invertebrates tested was Be > Sb > Ba based on the median effective concentration values for reproduction. These studies produced toxicological data that can contribute to the development of Eco-SSLs for Sb, Ba, and Be for soil invertebrates.

Survival and reproduction of Enchytraeus crypticus (Oligochaeta, Enchytraeidae) in a natural sandy loam soil amended with the nitro-heterocyclic explosives RDX and HMX

Munition manufacturing, disposal, testing, training and other operations at military sites produced elevated levels of explosives and related materials in soil. The effects of these persistent and highly mobile in the environment energetic materials on soil biota have not been sufficiently investigated. The U.S. Environmental Protection Agency (USEPA) in conjunction with stakeholders, is developing Ecological Soil Screening Level (Eco-SSL) benchmarks for ecological risk assessment (ERA) of contaminants at Superfund sites to identify those contaminants in soil that warrant additional evaluation in a Baseline ERA, and to eliminate those that do not. Eco-SSLs are developed from literature values whenever sufficient quantity and quality of data exist. Insufficient data were available on the toxicity of energetic compounds, RDX and HMX, to soil invertebrates, necessitating toxicity testing. Tests were conducted under conditions preferred for Eco-SSL benchmarks, using a Sassafras sandy loam soil that supports relatively high bioavailability of test compounds. Toxicity testing was performed using enchytraeid reproduction test (ISO/16387 2001) measuring adult survival and juvenile production by the potworm Enchytraeus crypticus in freshly amended soil and weathered/aged amended soil. RDX or HMX had no effect on adult survival in the definitive tests in all treatment concentrations. Juvenile production EC 2 0 values were 3,715 and 8,797 mg kg - 1 RDX in freshly amended and weathered/aged amended soils, respectively. Juvenile E. crypticus production was not adversely affected by exposure to HMX in freshly amended and weathered/aged amended soils. Juvenile production was stimulated in freshly amended soil up to 21,750 mg kg - 1 HMX. Results of these toxicity studies will be submitted to the Eco-SSL Task Group for quality control review, and pending approval will be included in the Eco-SSL database for Eco-SSL derivation.

Weathering and aging of 2,4,6-trinitrotoluene in soil increases toxicity to potworm Enchytraeus crypticus

Energetic materials are employed in a wide range of commercial and military activities and often are released into the environment. Scientifically based ecological soil-screening levels (Eco-SSLs) are needed to identify contaminant explosive levels in soil that present an acceptable ecological risk. Insufficient information for 2,4,6-trinitrotoluene (TNT) to generate Eco-SSLs for soil invertebrates necessitated toxicity testing. We adapted the standardized Enchytraeid Reproduction Test and selected Enchytraeus crypticus for these studies. Tests were conducted in Sassafras sandy loam soil, which supports relatively high bioavailability of TNT. Weathering and aging procedures for TNT amended to test soil were incorporated into the study design to produce toxicity data that better reflect the soil exposure conditions in the field compared with toxicity in freshly amended soils. This included exposing hydrated TNT-amended soils in open glass containers in the greenhouse to alternating wetting and drying cycles. Definitive tests showed that toxicity for E. crypticus adult survival and juvenile production was increased significantly in weathered and aged soil treatments compared with toxicity in freshly amended soil based on 95% confidence intervals. The median effect concentration and 20% effective concentration for reproduction were 98 and 77 mg/kg, respectively, for TNT freshly amended into soil and 48 and 37 mg/kg, respectively, for weathered and aged TNT soil treatments. These findings of increased toxicity to E. crypticus in weathered and aged TNT soil treatments compared with exposures in freshly amended soils show that future investigations should include a weathering and aging component to generate toxicity data that provide more complete information on ecotoxicological effects of energetic contaminants in soil.

A proteome-based assessment of the earthworm Eisenia fetida: response to chemical warfare agents in a sandy loam soil

Proteins are known to respond rapidly when an organism is exposed to a toxicant and this response is consistent and unique to the xenobiotics. Induced and repressed proteins together create a specific signature that can be used to identify a toxic agent and its concentration in the environmental matrix. The resulting protein expression signature for each agent will include a small number of the most potent proteins, which can be used to detect the presence and determine the concentration of a toxicant.We exposed the earthworm Eisenia fetida to a range of concentrations of chemical warfare agents (CWA) HD (mustard) and VX (nerve agent) in a natural sandy loam soil. Results demonstrated specificity of protein signatures in response to HD and VX exposure in soil. This study also showed that protein patterns were specific for different concentrations of HD or VX in otherwise clean agent-amended soil. Several proteins were induced in all HD or VX concentrations tested suggesting the potential of identifying the key proteins for use in detection of HD orVX presence in soil regardless of their concentrations. The robustness of the proteome-based response to chemical exposure by soil biota needs to be further investigated by including determinations of protein patterns for chemicals in different soil types and in the presence of additional chemical stressors using variety of soil invertebrate models.

Toxicities of dinitrotoluenes and trinitrobenzene freshly amended or weathered and aged in a sandy loam soil to Enchytraeus crypticus

Scientifically based ecological soil-screening levels are needed to identify concentrations of contaminant energetic materials (EMs) in soil that present an acceptable ecological risk at a wide range of military installations. Insufficient information regarding the toxicity of 2,4-dinitrotoluene (2,4-DNT), 2,6-dinitrotoluene (2,6-DNT), and 1,3,5-trinitrobenzene (TNB) to soil invertebrates necessitated toxicity testing. We adapted the standardized Enchytraeid Reproduction Test (International Standardization Organization 16387:2003) and selected Enchytraeus crypticus for these studies. Tests were conducted in Sassafras sandy loam soil, which supports relatively high bioavailability of nitroaromatic EMs. Weathering and aging procedures for EMs amended to test soil were incorporated into the study design to produce toxicity data that better reflect the soil exposure conditions in the field compared with toxicity in freshly amended soils. This included exposing hydrated, EM-amended soils in open glass containers in the greenhouse to alternating wetting and drying cycles. Definitive tests established that the order of EM toxicity to E. crypticus based on the median effect concentration values for juvenile production in either freshly amended or weathered and aged treatments was (from the greatest to least toxicity) TNB > 2,4-DNT > 2,6-DNT. Toxicity to E. crypticus juvenile production was significantly increased in 2,6-DNT weathered and aged soil treatments compared with toxicity in freshly amended soil, based on 95% confidence intervals. This result shows that future investigations should include a weathering and aging component to generate toxicity data that provide more complete information regarding ecotoxicological effects of energetic contaminants in soil.

Soil properties affect the toxicities of 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) to the enchytraeid worm Enchytraeus crypticus.

The authors investigated individual toxicities of 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) to the potworm Enchytraeus crypticus using the enchytraeid reproduction test. Studies were designed to generate ecotoxicological benchmarks that can be used for developing ecological soil-screening levels for ecological risk assessments of contaminated soils and to identify and characterize the predominant soil physicochemical parameters that can affect the toxicities of TNT and RDX to E. crypticus. Soils, which had a wide range of physicochemical parameters, included Teller sandy loam, Sassafras sandy loam, Richfield clay loam, Kirkland clay loam, and Webster clay loam. Analyses of quantitative relationships between the toxicological benchmarks for TNT and soil property measurements identified soil organic matter content as the dominant property mitigating TNT toxicity for juvenile production by E. crypticus in freshly amended soil. Both the clay and organic matter contents of the soil modulated reproduction toxicity of TNT that was weathered and aged in soil for 3 mo. Toxicity of RDX for E. crypticus was greater in the coarse-textured sandy loam soils compared with the fine-textured clay loam soils. The present studies revealed alterations in toxicity to E. crypticus after weathering and aging TNT in soil, and these alterations were soil- and endpoint-specific.

Controlled environment soil‐core microcosm unit for investigating fate, migration, and transformation of chemicals in soils

Abstract The controlled environment soil‐core microcosm unit (CESMU) methods embody a collection of techniques that began with soil sampling in the field and continued throughout the laboratory investigation of chemical fate, migration, and transformation in site‐specific soils; it was a cost‐effective investigative methodology that could be used to screen chemical materials before initiating high‐cost environmental field studies. Intact soil cores were collected in the field using a hydraulically controlled probe, delivering intact soil‐cores with minimal disturbance directly into high‐density polyethylene pipe (10.3‐cm ID). The inert polyethylene pipe was an effective hydrophobic barrier that remained an integral part of the soil‐core column, obviating subsequent transfers of soil. In the laboratory, each soil column was fitted with a porous ceramic plate and a polyethylene endcap containing fittings for teflon tubing, so that a tension could be applied at the bottom of each soil column (30–35 kPa) to m...

Selenium toxicity to survival and reproduction of Collembola and Enchytraeids in a sandy loam soil

We investigated the toxicity of selenium (Se) to the soil invertebrates Folsomia candida (Collembola) and Enchytraeus crypticus (potworm). Studies were designed to generate ecotoxicological benchmarks for developing ecological soil screening levels (Eco-SSLs) for risk assessments of contaminated soils. For the present studies, we selected Sassafras sandy loam, an aerobic upland soil with soil characteristics (low levels of clay and organic matter, soil pH adjusted from 5.2 to 7.1) that support high relative bioavailability of the anionic Se species that is typically found in aerobic soil. The Se was amended into soil as sodium selenate, subjected to weathering and aging using 21 d of alternating cycles of air-drying/rehydration to 60% of the water-holding capacity of the Sassafras sandy loam soil, under ambient greenhouse conditions. Effective concentrations at 20 and 50% (EC20 and EC50) levels for production of juveniles (reproduction) were 4.7 and 10.9 mg of Se/kg of soil (dry mass basis), respectively, for Collembola, and 4.4 and 6.2 mg/kg, respectively, for the potworms. The data enabled the derivation of toxicity benchmarks, contributing to the development of a soil invertebrate-based Eco-SSL of 4.1 mg/kg for Se. Environ Toxicol Chem 2018;37:846-853. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.