Alessandra C. Leri

Natural niche for organohalide-respiring Chloroflexi.

ABSTRACT The phylum Chloroflexi contains several isolated bacteria that have been found to respire a diverse array of halogenated anthropogenic chemicals. The distribution and role of these Chloroflexi in uncontaminated terrestrial environments, where abundant natural organohalogens could function as potential electron acceptors, have not been studied. Soil samples (116 total, including 6 sectioned cores) from a range of uncontaminated sites were analyzed for the number of Dehalococcoides-like Chloroflexi 16S rRNA genes present. Dehalococcoides-like Chloroflexi populations were detected in all but 13 samples. The concentrations of organochlorine ([organochlorine]), inorganic chloride, and total organic carbon (TOC) were obtained for 67 soil core sections. The number of Dehalococcoides-like Chloroflexi 16S rRNA genes positively correlated with [organochlorine]/TOC while the number of Bacteria 16S rRNA genes did not. Dehalococcoides-like Chloroflexi were also observed to increase in number with a concomitant accumulation of chloride when cultured with an enzymatically produced mixture of organochlorines. This research provides evidence that organohalide-respiring Chloroflexi are widely distributed as part of uncontaminated terrestrial ecosystems, they are correlated with the fraction of TOC present as organochlorines, and they increase in abundance while dechlorinating organochlorines. These findings suggest that organohalide-respiring Chloroflexi may play an integral role in the biogeochemical chlorine cycle.

Natural organobromine in marine sediments: New evidence of biogeochemical Br cycling

Received 22 January 2010; revised 6 July 2010; accepted 4 August 2010; published 24 November 2010. [1] Organobromine (Brorg) compounds, commonly recognized as persistent, toxic anthropogenic pollutants, are also produced naturally in terrestrial and marine systems. Several enzymatic and abiotic bromination mechanisms have been identified, as well as an array of natural Brorg molecules associated with various marine organisms. The fate of the carbon‐bromine functionality in the marine environment, however, remains largely unexplored. Oceanographic studies have noted an association between bromine (Br) and organic carbon (Corg) in marine sediments. Even so, there has been no direct chemical evidence that Br in the sediments exists in a stable form apart from inorganic bromide (Brinorg), which is widely presumed conservative in marine systems. To investigate the scope of natural Brorg production and its fate in the environment, we probed Br distribution and speciation in estuarine and marine sediments using in situ X‐ray spectroscopy and spectromicroscopy. We show that Brorg is ubiquitous throughout diverse sedimentary environments, occurring in correlation with Corg and metals such as Fe, Ca, and Zn. Analysis of sinking particulate carbon from the seawater column links the Brorg observed in sediments to biologically produced Brorg compounds that persist through humification of natural organic matter (NOM). Br speciation varies with sediment depth, revealing biogeochemical cycling of Br between organic and inorganic forms as part of the burial and degradation of NOM. These findings illuminate the chemistry behind the association of Br with Corg in marine sediments and cast doubt on the paradigmatic classification of Br as a conservative element in seawater systems.

Preservation of organic matter in marine sediments by inner-sphere interactions with reactive iron

Interactions between organic matter and mineral matrices are critical to the preservation of soil and sediment organic matter. In addition to clay minerals, Fe(III) oxides particles have recently been shown to be responsible for the protection and burial of a large fraction of sedimentary organic carbon (OC). Through a combination of synchrotron X-ray techniques and high-resolution images of intact sediment particles, we assessed the mechanism of interaction between OC and iron, as well as the composition of organic matter co-localized with ferric iron. We present scanning transmission x-ray microscopy images at the Fe L3 and C K1 edges showing that the organic matter co-localized with Fe(III) consists primarily of C=C, C=O and C-OH functional groups. Coupling the co-localization results to iron K-edge X-ray absorption spectroscopy fitting results allowed to quantify the relative contribution of OC-complexed Fe to the total sediment iron and reactive iron pools, showing that 25–62% of total reactive iron is directly associated to OC through inner-sphere complexation in coastal sediments, as much as four times more than in low OC deep sea sediments. Direct inner-sphere complexation between OC and iron oxides (Fe-O-C) is responsible for transferring a large quantity of reduced OC to the sedimentary sink, which could otherwise be oxidized back to CO2.

Abiotic Bromination of Soil Organic Matter

Biogeochemical transformations of plant-derived soil organic matter (SOM) involve complex abiotic and microbially mediated reactions. One such reaction is halogenation, which occurs naturally in the soil environment and has been associated with enzymatic activity of decomposer organisms. Building on a recent finding that naturally produced organobromine is ubiquitous in SOM, we hypothesized that inorganic bromide could be subject to abiotic oxidations resulting in bromination of SOM. Through lab-based degradation treatments of plant material and soil humus, we have shown that abiotic bromination of particulate organic matter occurs in the presence of a range of inorganic oxidants, including hydrogen peroxide and assorted forms of ferric iron, producing both aliphatic and aromatic forms of organobromine. Bromination of oak and pine litter is limited primarily by bromide concentration. Fresh plant material is more susceptible to bromination than decayed litter and soil humus, due to a labile pool of mainly aliphatic compounds that break down during early stages of SOM formation. As the first evidence of abiotic bromination of particulate SOM, this study identifies a mechanistic source of the natural organobromine in humic substances and the soil organic horizon. Formation of organobromine through oxidative treatments of plant material also provides insights into the relative stability of aromatic and aliphatic components of SOM.

Sample thickness and quantitative concentration measurements in Br K-edge XANES spectroscopy of organic materials

While XANES spectroscopy is an established tool for quantitative information on chemical structure and speciation, elemental concentrations are generally quantified by other methods. The edge step in XANES spectra represents the absolute amount of the measured element in the sample, but matrix effects and sample thickness complicate the extraction of accurate concentrations from XANES measurements, particularly at hard X-ray energies where the X-ray beam penetrates deeply into the sample. The present study demonstrates a method of quantifying concentration with a detection limit approaching 1 mg kg(-1) using information routinely collected in the course of a hard X-ray XANES experiment. The XANES normalization procedure unambiguously separates the signal of the absorber from any source of background. The effects of sample thickness on edge steps at the bromine K-edge were assessed and an empirical correction factor for use with samples of variable mass developed.

Formation of organochlorine by-products in bleached laundry

Laundering fabrics with chlorine bleach plays a role in health and hygiene as well as aesthetics. However, laundry bleaching may create chlorinated by-products with potentially adverse human health effects. Studies have shown that toxic chlorinated gases are produced in the headspace of washing machines when hypochlorite-containing bleach is used. Laundry bleaching has also been implicated in contributing dissolved organochlorine to municipal wastewater. However, there have been no reports of organochlorines produced and retained in fabric as a result of laundry bleaching. We have used a chlorine-specific X-ray spectroscopic analysis to demonstrate the formation of organochlorine by-products in cotton fabrics laundered with chlorine bleach under typical household conditions. Organochlorine formation increases at higher wash temperature. At least two pools of organochlorine are produced in bleached fabric: a labile fraction that diminishes over several months of storage time as well as a more stable fraction that persists after more than 1 year. Our results also suggest that residual hypochlorite remains in fabric after laundering with bleach, presenting the possibility of direct and sustained dermal contact with reactive chlorine. This study provides a first step toward identifying a new risk factor for elevated organochlorine body burdens in humans.

Particulate organohalogens in edible brown seaweeds

Brown algae, rich in antioxidants and other bioactive compounds, are important dietary seaweeds in many cultures. Like other marine macroalgae, brown seaweeds are known to accumulate the halogens iodine and bromine. Comparatively little is known about the chemistry of chlorine in seaweeds. We used synchrotron-based X-ray absorption spectroscopy to measure total non-volatile organochlorine and -bromine in five edible brown seaweeds: Laminaria digitata, Fucus vesiculosus, Pelvetia canaliculata, Saccharina latissima, and Undaria pinnatifida. Organochlorine concentrations range from 120 to 630 mg·kg-1 dry weight and organobromine from 150 to 360 mg·kg-1, comprising mainly aromatic organohalogens in both cases. Aliphatic organochlorine exceeds aliphatic organobromine but is positively correlated with it among the seaweeds. Higher organochlorine levels appear in samples with more lipid moieties, suggesting lipid chlorination as a possible formation pathway. Particulate organohalogens are not correlated with antioxidant activity or polyphenolic content in seaweed extracts. Such compounds likely contribute to organohalogen body burden in humans and other organisms.