A Rapid, Fluorescence-Based Field Screening Technique for Organic Species in Soil and Water Matrices.

Real-time detection of hydrocarbon contaminants in the environment presents analytical challenges because traditional laboratory-based techniques are cumbersome and not readily field portable. In the current work, a method for rapid and semi-quantitative detection of organic contaminants, primarily crude oil, in natural water and soil matrices has been developed. Detection limits in the parts per million and parts per billion were accomplished when using visual and digital detection methods, respectively. The extraction technique was modified from standard methodologies used for hydrocarbon analysis and provides a straight-forward separation technique that can remove interference from complex natural constituents. For water samples this method is semi-quantitative, with recoveries ranging from 70 % to 130 %, while measurements of soil samples are more qualitative due to lower extraction efficiencies related to the limitations of field-deployable procedures.

Effect size of delayed freezing, diurnal variation, and hindgut location on the mouse fecal microbiome.

Practical considerations in fecal sample collection for microbiome research include time to sample storage, time of collection, and hindgut position during terminal collections. Here, parallel experiments were performed to investigate the relative effect of these factors on microbiome composition in mice colonized with two different vendor-origin microbiomes. 16S rRNA amplicon sequencing of immediately flash-frozen feces showed no difference in alpha or beta diversity compared to samples incubated up to 9 h at room temperature. Samples collected in the morning showed greater alpha diversity compared to samples collected in the afternoon. While a significant effect of time was detected in all hindgut regions, the effect increased from cecum to distal colon. This study highlights common scenarios in microbiome research that may affect outcome measures of microbial community analysis. However, we demonstrate a relatively low effect size of these technical factors when compared to a primary experimental factor with large intergroup variability.

The contribution of maternal oral, vaginal, and gut microbiota to the developing offspring gut.

There is limited understanding of how the microbiota colonizing various maternal tissues contribute to the development of the neonatal gut microbiota (GM). To determine the contribution of various maternal microbiotic sites to the offspring microbiota in the upper and lower gastrointestinal tract (GIT) during early life, litters of mice were sacrificed at 7, 9, 10, 11, 12, 14, and 21 days of age, and fecal and ileal samples were collected. Dams were euthanized alongside their pups, and oral, vaginal, ileal, and fecal samples were collected. This was done in parallel using mice with either a low-richness or high-richness microbiota to assess the consistency of findings across multiple microbial compositions. Samples were analyzed using 16S rRNA amplicon sequencing. The compositional similarity between pup and dam samples were used to determine the contribution of each maternal source to the composition of the neonate fecal and ileal samples at each timepoint. As expected, similarity between neonate and maternal feces increased significantly over time. During earlier time-points however, the offspring fecal and ileal microbiotas were closer in composition to the maternal oral microbiota than other maternal sites. Prominent taxa contributed by the maternal oral microbiota to the neonate GM were supplier-dependent and included Lactobacillus spp., Streptococcus spp., and a member of the Pasteurellaceae family. These findings align with the microbial taxa reported in infant microbiotas, highlighting the translatability of mouse models in this regard, as well as the dynamic nature of the GM during early life.

Methods for simultaneous determination of legacy and insensitive munition (IM) constituents in aqueous, soil/sediment, and tissue matrices.

Currently, no standard method exists for analyzing insensitive munition (IM) compounds in environmental matrices, with or without concurrent legacy munition compounds, resulting in potentially inaccurate determinations. The primary objective of this work was to develop new methods of extraction, pre-concentration, and analytical separation/quantitation of 17 legacy munition compounds along with several additional IM compounds, IM breakdown products, and other munition compounds that are not currently included in U. S. Environmental Protection Agency (EPA) Method 8330B. The eight additional compounds included were nitroguanidine, 3-nitro-1,2,4-triazol-5-one, picric acid, 2,4-dinitroanisole, 2,4-dinitrophenol, 2-nitrophenol, 4-nitrophenol, and new surrogate ortho-nitrobenzoic acid (o-NBA). Analytical methods were developed to enable sensitive, simultaneous detection and quantitation of the 24 IM and legacy compounds, including two orthogonal high-performance liquid chromatography (HPLC) column separations with either ultraviolet (UV) or mass spectrometric (MS) detection. Procedures were developed for simultaneous extraction of all 24 analytes and two surrogates (1,2-dinitrobenzene, 1,2-DNB; o-NBA) from high- and low-level aqueous matrices and solid matrices, using acidification, solid phase extraction (SPE), or solvent extraction (SE), respectively. For low-level aqueous samples extracted by SPE, all compounds were recovered within current Department of Defense Quality Systems Manual (DoD QSM) Ver5.3 accepted limits for aqueous samples analyzed by EPA Method 8330B (57-135%), except NQ, which was consistently recovered at approximately 50%. Likewise, all compounds were recovered from six geographically/geochemically unique soil types within current QSM accepted limits for solid samples analyzed by EPA Method 8330B (64-135%). Further, the majority of compounds were recovered from four tissue types within current limits for solids, with generally low recovery only for Tetryl (from 4 to 62%). A preparatory chromatographic interference removal procedure was adapted for tissue extracts, as various analytical interferences were observed for all studied tissue types.

Radical Cation/Radical Reactions: A Fourier Transform Ion Cyclotron Resonance Study of Allyl Radical Reacting with Aromatic Radical Cations.

A method for the study of reactions of open-shell neutrals (radicals) and radical cations is described. Pyrolysis (25-1500 degrees C) of thermally labile compounds, such as, 1,5-hexadiene via a Chen nozzle yields a seeded beam of reactive species in helium. The pyrolysis products are then analyzed by electron ionization (EI) or reacted with stored ions. Electron ionization of the pyrolysis products of 1,5-hexadiene shows that both the allyl radical and allene are generated. Reactions of benzene radical cations and the pyrolysis products of 1,5-hexadiene result in carbon-carbon bond formation. Those reactions of allyl radical with the benzene radical cation yield the C(7)H(7) (+) ion of m/z 91, permitting an unusual entry into arenium ions. The reaction of allene with benzene radical cation in contrast yields C(9)H(10) (+). and C(9)H(9) (+).

Accumulation of 2,4-dinitroanisole in the earthworm Eisenia fetida from chemically spiked and aged natural soils.

An initiative within the US military is targeting the replacement of traditional munitions constituents with insensitive munitions to reduce the risk of accidental detonation. The bioavailability and bioaccumulative potential of the insensitive munitions compound 2,4-dinitroanisole (DNAN) to Eisenia fetida was assessed in soils with different geochemical characteristics. Prior to exposure, soils were chemically spiked with DNAN and aged for 1 wk or 29 wk. Transformation products 2- and 4-amino-nitroanisole (2A-4NAN and 4A-2NAN) occurred in aged soils and their porewater but never at concentrations higher than the residual DNAN. The sum of DNAN, 2A-4NAN, and 4A-2NAN (sumDNAN) in soil decreased with aging, likely by irreversible binding. Both clay and organic matter contents of the soil appeared to affect the bioavailability of DNAN. The sumDNAN body residues of earthworms approached apparent steady state after 1 d and remained relatively constant through to day 7. Higher concentrations of 2A-4NAN and 4A-2NAN measured in worm tissues relative to those in soil suggest reductive transformation of DNAN in the tissues. Mean bioaccumulation factors (ratio of tissue to soil concentrations) varied from 1.2 to 4.3, whereas mean bioconcentration factors (ratio of tissue to porewater concentrations) ranged from 1.4 to 3.2. Porewater seems to play a significant role in the accumulation of DNAN in earthworms, consistent with equilibrium partitioning theory. The concentration of DNAN in soil porewater could serve as an indicator of bioavailability as well as a predictor of the concentration of that compound in earthworms. Environ Toxicol Chem 2016;35:1835-1842. Publlished 2015 SETAC. This article is a US Government work, and as such, is in the public domain in the United States of America.

Inter- and intraspecies chemical sensitivity: a case study using 2,4-dinitroanisole.

Insensitive munitions offer increased safety because of their "insensitivity" to unintended detonation relative to historically used formulations such as 2,4,6-trinitrotoluene (TNT). Dinitroanisole (DNAN) is an insensitive munition constituent, and its solubility and stability warrant investigations of potential toxicological hazard related to manufacturing discharges and training ranges. Although ecotoxicology data are available for other insensitive munition constituents, few data are available for DNAN. In the present study, acute and chronic exposures of a fish (Pimephales promelas) and 2 cladocerans (Ceriodaphnia dubia, Daphnia pulex) were conducted. The 50% lethal concentration (LC50) values of DNAN ranged from 14.2 mg/L to 42.0 mg/L, depending on species. In chronic exposures, fish survival (LC50 = 10.0 mg/L) was more sensitive than cladoceran survival (LC50 = 13.7 to >24.2 mg/L). However, cladoceran reproduction was equally or more sensitive to DNAN (50% inhibition values 2.7-10.6 mg/L, depending on species) than fish endpoints. Daphnia pulex was the most sensitive species, with only slight differences between the 3 populations tested. Although the aquatic toxicity of DNAN was lower than previously reported in the literature for TNT, future research is needed to determine the potential synergistic toxicity of all the constituents in insensitive munition mixtures and the implications of photo-oxidation.

Algal growth stimulation and toxicity in response to exposure to the new insensitive military high-nitrogen energetic triaminoguanidinium-1-methyl-5-nitriminotetrazolate.

Triaminoguanidinium-1-methyl-5-nitriminotetrazolate (TAG-MNT) is a nitrogen-rich energetic compound being developed as a potential component of insensitive munition formulations. The purpose of the present study was to assess the toxicity of TAG-MNT to the green alga Pseudokirchneriella subcapitata as well as to determine whether the high N content of TAG-MNT could result in increased algal growth in aquatic systems and potentially contribute to eutrophication using a 96-h algal growth bioassay in N-limited test media. Results were compared with algal exposures to current-use energetics 2,4,6-trinitrotoluene (TNT) and royal demolition explosive (RDX). The TNT exposure resulted in a lowest-observed-adverse-effect concentration (LOAEC) for algal growth of 1.72 mg/L and a 50% inhibition concentration (IC50) and 95% confidence limits of 0.972 mg/L (0.955, 0.973). The RDX algal growth LOAEC was 0.10 mg/L, and the RDX IC50 was 0.635 (0.416, 0.875). Neither TNT nor RDX exposure resulted in stimulation of algal growth. In repeated testing, TAG-MNT exposure resulted in LOAECs of 0.55 and 5.20 mg/L. Stimulation of algal growth was observed at 0.06 mg/L at a mean increase of 163.2% (± 71.7) relative to the control in TAG-MNT test A and at the 0.005 mg/L treatment at a mean increase of 174.3% (± 59.9) in TAG-MNT test B. The authors' results indicate the potential for high-N energetics to significantly stimulate algal growth at low concentrations in N-limited systems.