Some aspects of the ecophysiology of Tubificoides benedii and ultrastructural observations on endocuticular bacteria

Tubificoides benedii is regularly found in sulphide-rich sediments with extremely low oxygen tensions and can tolerate anaerobic conditions for several days. Although the anaerobic energy production of marine invertebrates has been well studied, almost nothing is known about the anaerobic metabolism of marine oligochaetes. Preliminary results after measuring end-products during anaerobic incubation show that in contrast to all previously examined marine facultative anaerobe invertebrates T. benedii degrades malate during anaerobiosis. Also, the concentration of free amino acids is extremely low for a marine organism. Low levels of free amino acids could be concomitant with malate utilization: the utilization of the amino acid aspartate (as observed in all other examined marine invertebrates) seems to be excluded by the low concentrations of aspartate and other amino acids in T. benedii.The physiological lab studies were supplemented by ecological investigations in the field and laboratory on the vertical distribution of T. benedii. 90% of the population was always found within the first few cm below the sediment surface. Aquarium observations showed that the posterior end of the worm projects above the sediment surface, where it slowly waves back and forth. This behavior points towards an intestinal respiration. The described orientation, an intestinal respiration and anaerobic energy production could be advantageous in sulphide-rich sediments where O₂ only penetrates a few mm into the sediment. The worm can easily inhabit the first three to four cm by holding its tail in the upper oxygenated sediment and water. Here it would be able to feed on the rich quantities of bacteria at the anoxic-oxic interface and yet still keep up an aerobic metabolism. In addition, its ability to produce energy anaerobically would allow T. benedii to dwell in deeper anoxic sediments for limited periods of time or to survive complete O₂ absence that could develop during low tide.The posterior ends of T. benedii found in a sulphide-rich habitat in the German Wadden Sea were covered with filamentous epibacteria (Dubilier, 1986). Electron microscopy showed that the bacteria were anchored in the cuticle. The association is apparently not pathogenic whereas positive forms of interaction can be envisioned.     

Sulfide Tolerance and Detoxification in Arenicola marina and Sipunculus nudus

SYNOPSIS. The lugworm Arenicola marina and the peanut worm Sipunculusnudus both inhabit intertidal flats. Sulfide concentrationsin the pore water of the sediment can be as high as 340 µMin the habitat of A. marina. Sulfide levels in the burrow waterand in the coelomic fluid increase during low tide. In contrast,S. nudus is only rarely exposed to sulfide. Neither A. marinanor S. nudus is able to exclude sulfide completely from itsbody. However, both worms accumulate only 15 to 40% respectivelyof the external sulfide concentration during hypoxic 1 mM sulfideincubations. During normoxia, sulfide accumulation is more thanten-fold lower than under hypoxic conditions in both species.When oxygen is available, sulfide is oxidized to thiosulfate.The oxidation rate is about 5 times higher in A. marina thanin S. nudus. A major component of sulfide oxidation is due tothe mitochondria of both species. Mitochondrial sulfide oxidationin the tissues of A. marina is insensitive to low cyanide andto high sulfide concentrations, but can be inhibited by salicylhydroxamicacid. The existence of an alternative terminal oxidase is suggested.When the sulfide level in the body of A. marina and S. nudusincreases above a toxic level, both species survive by switchingto an anaerobic metabolism.     

Multiple Mechanisms Provide Tolerance to Environmental Sulfide in Urechis caupo

SYNOPSIS. During tidal exposure the echiuran worm Urechis caupoencounters toxic sulnde in its burrow microhabitat on the marinemudflat. Although sulnde freely penetrates the two respiratoryepithelia (the body wall and hindgut) and tissue cytochromeoxidases are extremely sulnde sensitive, the worm toleratessulnde concentrations which exceed environmental exposures.Sulnde tolerance results from a suite of sulnde detoxificationmechanisms that extend from the coelomic fluid to the epithelialsurfaces. The coelomic fluid is rich in hemoglobin and hematin,both of which bind sulfide and catalyze the oxidation of sulfideto thiosulfate in vitro. Peripheral defense mechanisms responsiblefor protection of aerobic epithelia from sulfide poisoning mayinclude mucus secretion, sulfide oxidation by symbiotic bacteriaand/or specialized organelles termed SOBs, and sloughing ofsulfide damaged cells. Thiosulfate, the principal detoxificationendproduct produced in vivo, is eliminated primarily by diffusionacross the hindgut, and sulfur metabolites may be sequesteredand eliminated by the anal sacs. Based on a hypothetical model,we conclude that sulfide tolerance in U. caupo is due primarilyto the sulfide oxidation activity of the coelomic fluid andthat the specialized integument and hindgut protect the metabolicallyactive, sulfide-exposed epithelial cells. These sulfide detoxificationmechanisms may allow populations of this filter-feeding wormto exploit nutrient-rich, sulfidic environments.     

PRODUCTION OF HYDROGEN SULFIDE FROM SULFITE BY A COPPER-ADAPTED YEAST

Activity of hydrogen sulfide production from sulfite was studiedusing a copper-resistant yeast strain (R), its parent strain(P), and the culture of R in the medium without copper addition(R(0)). More hydrogen sulfide was produced under aerobic conditionthan under anaerobic condition. Sulfide producing activity wasin the order of R(0)>P>R under either condition. Stationaryphase cells produced more sulfide than logarithmic phase cellswhen cultured without copper, while the reverse was the casewith R, cultured in copper medium. Sulfide production was inhibitedby high concentrations of sulfite and by salicylaldoxime. Differencein the pathway from sulfite to sulfide was suggested betweenthe resistant strain (R and R(0)) and P in that the former wasmore sensitive to these inhibitors. ¹ Present address: Research Reactor Institute, Kyoto University,Kumatori-cho, Sennan-gun, Osaka     

Cellular Energetics of Animals from High Sulfide Environments

SYNOPSIS.Mitochondrial ATP production is influenced by manyfactors, including the adenylate status of the cell, the supplyof reducing equivalents to the electron transport chain, thesupply of oxygen to cytochrome oxidase, and the demand for ATPto do cellular work. Hydrogen sulfide, which is naturally producedin marine sediments, is a poison of aerobic ATP production mainlybecause it inhibits cytochrome oxidase in the electron transportchain. However, most animals from high sulfide environmentsexhibit aerobic respiration, and may avoid sulfide poisoningwith detoxification reactions that may be useful sources ofenergy. Sulfide stimulates ADP phosphorylation in mitochondriaisolated from gills of Solemya reidi, a sulfide-oxidizing symbiont-harboringbivalve, and a P/O ratio near unity indicates that electronsfrom sulfide enter the electron transport chain at the levelof cytochrome c. Current investigations into the effects ofsulfide on oxygen consumption rate, ATP level, cytochrome reductionstate and ciliary beat frequency of symbiont-free gills of themussels Geukensia demissa and Mytilus edulis indicate that animalsfrom high sulfide environments may gain sufficient energy fromsulfide oxidation to support cellular work.     

Cooccurrence of Aerobic and Anaerobic Methane Oxidation in the Water Column of Lake Plußsee

Dissolved methane was investigated in the water column of eutrophic Lake Plußsee and compared to temperature, oxygen, and sulfide profiles. Methane concentrations and δ-¹³C signatures indicated a zone of aerobic methane oxidation and additionally a zone of anaerobic methane oxidation in the anoxic water body. The latter coincided with a peak in hydrogen sulfide concentration. High cell numbers of aerobic and anaerobic methane-oxidizing microorganisms were detected by fluorescence in situ hybridization (FISH) or the more sensitive catalyst-amplified reporter deposition-FISH, respectively, in these layers.     

Degradation of sulfide by dehaloperoxidase-hemoglobin from <Emphasis Type="Italic">Amphitrite ornata</Emphasis>

Dehaloperoxidase-hemoglobin (DHP) is a unique multifunctional enzyme with a globin fold. The enzyme serves as the respiratory hemoglobin for the marine worm Amphitrite ornata and has been shown to catalyze the conversion of highly toxic trihalophenols to dihaloquinones as a detoxification function for the organism. Given the simplicity of the structure of A. ornata, it is entirely possible that DHP may play an even more general role in detoxification of the organism from sulfide commonly found in the coastal estuaries where A. ornata thrives. Comparison of DHP with other sulfide-binding hemoglobins shows that DHP possesses several distal cavity structural properties, such as an aromatic cage and a hydrogen-bond-donor amino acid (His55), that facilitate sulfide binding. Furthermore, a complete reduction of the ferric heme occurs after sulfide exposure under aerobic or anaerobic conditions to yield either the oxy or the deoxy ferrous states of DHP, respectively. Oxidation of sulfide by the heme leads to sulfur products that are less toxic to A. ornata. This proposed new function for DHP relies on the highly flexible distal His55 for deprotonation of the bound hydrogen sulfide, similar to H₂O₂ activation of the peroxidase function, and provides further support for the importance of the flexibility of the distal His55 in this novel globin.     

Cooccurrence of aerobic and anaerobic methane oxidation in the water column of Lake Plusssee

Dissolved methane was investigated in the water column of eutrophic Lake Plusssee and compared to temperature, oxygen, and sulfide profiles. Methane concentrations and delta-13C signatures indicated a zone of aerobic methane oxidation and additionally a zone of anaerobic methane oxidation in the anoxic water body. The latter coincided with a peak in hydrogen sulfide concentration. High cell numbers of aerobic and anaerobic methane-oxidizing microorganisms were detected by fluorescence in situ hybridization (FISH) or the more sensitive catalyst-amplified reporter deposition-FISH, respectively, in these layers.     

Impacts of reduced sulfur components on active and resting ammonia oxidizers

While there has been significant research on the nature and extent of the impact of inhibitory reduced sulfur with respect to anaerobic (e.g., methanogenic and sulfidogenic) microbial systems, only limited study has yet been conducted on the comparable effects of soluble sulfides which might occur within aerobic wastewater treatment systems. Admittedly, aerobic reactors would not normally be considered conducive to the presence of reduced sulfur constituents, but there do appear to be a number of processing scenarios under which related impacts could develop, particularly for sensitive reactions like nitrification. Indeed, the following scenarios might well involve elevated levels of reduced sulfur within an aerobic reactor environment: (1) mixed liquor recycle back through sulfide-generating anaerobic zones (e.g., in conjunction with biological nutrient removal processes, etc.), (2) high-level side-stream sulfide recycle via sludge digestion, etc., back to aerobic reactors, and (3) high-level influent sulfide inputs to wastewater treatment facilities via specific industrial, septage, etc., streams. The objective of this study was, therefore, to determine the subsequent metabolic impact of soluble sulfide under aerated and unaerated conditions, focusing in particular on ammonia-oxidizing bacteria due to their critical first-step role with nitrification. The obtained results indicated that, under catabolically active conditions, cultures of ammonia oxidizers were extremely sensitive to the presence of sulfide. At total soluble sulfide concentrations of 0.25 mg l^–1 S, active ammonia oxidation was completely inhibited. However, immediately following the removal of this soluble sulfide presence, ammonia oxidation started to recover; and it continued to improve over the next 24 h. Similar sulfide impact tests conducted with inactive ammonia oxidizers exposed during anaerobic conditions, albeit at higher dosage levels, also revealed that their subsequent aerobic activity would correspondingly be retarded. These results indicated that, after sulfide exposure under unaerated conditions, subsequent aerobic oxidative activity rates rapidly decreased as the soluble sulfide exposure was increased from 0.5 gm l^–1 S to 5 mg l^–1 S and that further reductions in this activity progressively developed as the concentration was increased to 200 mg l^–1 S. The recovery following unaerated exposure to sulfide was significantly higher at pH 7, as compared with pH 8, and although the specific nature of this variation was not established, a hypothetical explanation appeared warranted.     

Solubility of cadmium and cobalt in a post-oxic or sub-oxic sediment suspension

A sediment sample with high organic matter and trace metal content was suspended in synthetic river water for four weeks under an inert gas atmosphere. Subsequently, the anaerobic suspension was reoxidized by bubbling air through it. The concentrations of dissolved oxygen, sulfide, ferrous iron, manganese, cadmium, cobalt and the pH-value were measured at close time intervals during the anaerobic incubation. The anaerobic suspension was a post-oxic or sub-oxic environment with oxygen and total sulfide concentrations less than 1 µmole 1^–1. Concentrations of dissolved ferrous iron and manganese were 50–150 µmole 1^–1 and 5–30 µmole 1^–1, respectively. The total sulfide concentration was measured using a sensitive voltammetric technique, with a detection limit of 1 nmole 1^–1. A sequential extraction procedure was applied to two sediment samples taken at the end of the anaerobic incubation and after one week of reoxidation. The extractions indicated that cadmium was bound in sulfide minerals under post-oxic conditions. Thermodynamic equilibrium calculations revealed that the concentrations of dissolved cobalt in the post-oxic suspension were limited by the precipitation of cobalt sulfide minerals.     

Alvinellids and Sulfides at Hydrothermal Vents of the Eastern Pacific: A Review

SYNOPSIS. At Eastern Pacific hydrothermal vents, alvinellidpolychaetes are among the first metazoans to colonize newlyformed surfaces of sulfide chimneys. In this environment ofrapid mineral precipitation, alvinellids are confronted by steepphysico-chemical gradients and high temporalvariability. Thispaper examines the interaction of alvinellids with chimney mineralizationprocesses and then reviews what is known of mechanisms thatcould enable these worms to deal with potentially toxic levelsof sulfide in their environment. Studies of sulfide chimneysconsistently show mineralogy to be locally modified around alvinellidtubes. This may be linked to sulfide oxidation products thataccumulate in tube material or to the circulation of seawaterthrough the tube. At high worm densities, these local effectsmay have a significant influence on larger scale sulfide accretionprocesses that determine chimney morphology. Alvinellid polychaetesmay have several lines of defense against sulfide. Tubes andmucous layers could act as passive barriers to reduce inwarddiffusion of sulfide across posterior surfaces. Colonizationof epidermal and tube surfaces by bacteria that might be sulfideoxidizing, could create an active external barrier in some species.Sulfide oxidation by tissue homogenates has been demonstratedin two Paralvinella species, where it may serve to protect oxidativerespiration from sulfide entering worm tissues. Sulfide bindingin blood has not been studied in any of the alvinellids.     

The Effects of Kinetin and Gibberellin on the Germination of Dehusked Seeds of Indica and Japonica Rice (Oryza sativaL.) under Anaerobic and Aerobic Conditions

The effects of kinetin and gibberellin were examined under anaerobicconditions (0% oxygen) and aerobic conditions (20% oxygen) onthe germination of dehusked seeds of indica and japonica ricecultivars that had been harvested at different times duringthe formation of seeds. Surjamkhi was used as a representativeof deep dormant indica cultivars and Assam IV as a less dormantindica cultivar. Sasanishiki was used as the japonica rice cultivar.Both phytohormones were applied at a concentration of 10^-3Mwhichproved to have the greatest stimulatory effect in preliminarywork at concentrations of 10^-3–10^-5M. Under aerobic conditions,inhibition of germination by dehusking of Sasanishiki seedsthat had been harvested either 30 or 60 d after anthesis wasovercome by kinetin and all seeds germinated. Complete germinationinduced by kinetin under aerobic conditions was also achievedwith the dehusked seeds of the indica rice cultivar Assam IVthat had been harvested on two occasions and of Surjamkhi thathad been harvested 28 d after anthesis. In contrast, germinationof dehusked japonica seeds stimulated by anaerobiosis was inhibitedby kinetin. The stimulatory effects of gibberellin on the germinationof indica and japonica rice seeds were observed under aerobicand anaerobic conditions. Under anaerobic conditions, the responsesof dehusked indica and japonica rice seeds to kinetin and gibberellindiffered, being negative with kinetin and positive with gibberellin.Under aerobic conditions, the stimulatory effects of kinetinon germination of dehusked seeds were greater than those ofgibberellin. Thus, treatment with kinetin appears to be usefulfor breaking the considerable dormancy commonly observed inthe dehusked seeds of indica rice. Mechanisms are proposed toexplain the stimulatory effects of these phytohormones on thegermination of dehusked seeds of indica and japonica rice underaerobic and anaerobic conditions. Rice; Oryza sativaL.; seed germination; dehusking treatment; gibberellin; indica; japonica; kinetin; oxygen; dormancy; germination inhibition; seed formation     

Effect of Oxygen Radicals and Peroxide on Survival After Ultraviolet Irradiation and Liquid Holding Recovery of Bacteroides fragilis

The survival of Bacteroides fragilis cells after far-ultraviolet irradiation under anaerobic and aerobic conditions and the liquid holding recovery under aerobic conditions were not affected by peroxide or quenchers of toxic oxygen derivatives.     

Anaerobic versus Aerobic Degradation of Dimethyl Sulfide and Methanethiol in Anoxic Freshwater Sediments

Degradation of dimethyl sulfide and methanethiol in slurries prepared from sediments of minerotrophic peatland ditches were studied under various conditions. Maximal aerobic dimethyl sulfide-degrading capacities (4.95 nmol per ml of sediment slurry · h⁻¹), measured in bottles shaken under an air atmosphere, were 10-fold higher than the maximal anaerobic degrading capacities determined from bottles shaken under N₂ or H₂ atmosphere (0.37 and 0.32 nmol per ml of sediment slurry · h⁻¹, respectively). Incubations under experimental conditions which mimic the in situ conditions (i.e., not shaken and with an air headspace), however, revealed that aerobic degradation of dimethyl sulfide and methanethiol in freshwater sediments is low due to oxygen limitation. Inhibition studies with bromoethanesulfonic acid and sodium tungstate demonstrated that the degradation of dimethyl sulfide and methanethiol in these incubations originated mainly from methanogenic activity. Prolonged incubation under a H₂ atmosphere resulted in lower dimethyl sulfide degradation rates. Kinetic analysis of the data resulted in apparent K_m values (6 to 8 μM) for aerobic dimethyl sulfide degradation which are comparable to those reported for Thiobacillus spp., Hyphomicrobium spp., and other methylotrophs. Apparent K_m values determined for anaerobic degradation of dimethyl sulfide (3 to 8 μM) were of the same order of magnitude. The low apparent K_m values obtained explain the low dimethyl sulfide and methanethiol concentrations in freshwater sediments that we reported previously. Our observations point to methanogenesis as the major mechanism of dimethyl sulfide and methanethiol consumption in freshwater sediments.     

Suspension feeding and kleptoparasitism within the genus Trichotropis (Gastropoda: Capulidae)

The marine gastropod Trichotropis cancellata is a facultativekleptoparasite, either suspension feeding or parasitically stealingfood from tube-dwelling polychaete worms. To determine whetherconclusions drawn from long-term studies in the San Juan Islands,Washington, about the relative importance of suspension feedingand kleptoparasitism can be applied generally to T. cancellataacross its biogeographic range, I expanded earlier studies toAlaska and British Columbia. Kleptoparasitism is pervasive throughoutthe range of T. cancellata, occurring with equal frequency throughoutthe areas studied. The average density and size of worm hostsare relatively constant across this range. Snail and worm densitiesare not significantly correlated at the larger scale of site(averaged over nearby sampling locations clustered around acity), but are correlated at the smaller local scale (withina sampling location). Larger worms do not support more snails.The abundance of uninfected worm hosts is usually not limiting,except potentially in some sampling locations in southwest Alaskawhere the use of a novel host (a holothurian) may be the resultof low densities of uninfected worms. Additionally, I documentedthe feeding behaviours of other trichotropid species in theseregions. Trichotropis conica is the second confirmed kleptoparasitewithin the genus Trichotropis, with kleptoparasitism as frequentin this species as in T. cancellata. Like T. cancellata, allsizes observed of T. conica are kleptoparasites. On the otherhand, Trichotropis insignis is an obligate suspension feeder.Further studies are needed to determine exactly how many timesthis behaviour has arisen and been lost in Capulidae and relatedfamilies. (Received 31 May 2007; accepted 19 October 2007)     

Light-Dependent Sulfide Oxidation in the Anoxic Zone of the Chesapeake Bay Can Be Explained by Small Populations of Phototrophic Bacteria

Microbial sulfide oxidation in aquatic environments is an important ecosystem process, as sulfide is potently toxic to aerobic organisms. Sulfide oxidation in anoxic waters can prevent the efflux of sulfide to aerobic water masses, thus mitigating toxicity. The contribution of phototrophic sulfide-oxidizing bacteria to anaerobic sulfide oxidation in the Chesapeake Bay and the redox chemistry of the stratified water column were investigated in the summers of 2011 to 2014. In 2011 and 2013, phototrophic sulfide-oxidizing bacteria closely related to Prosthecochloris species of the phylum Chlorobi were cultivated from waters sampled at and below the oxic-anoxic interface, where measured light penetration was sufficient to support populations of low-light-adapted photosynthetic bacteria. In 2012, 2013, and 2014, light-dependent sulfide loss was observed in freshly collected water column samples. In these samples, extremely low light levels caused 2- to 10-fold increases in the sulfide uptake rate over the sulfide uptake rate under dark conditions. An enrichment, CB11, dominated by Prosthecochloris species, oxidized sulfide with a K_s value of 11 μM and a V_max value of 51 μM min⁻¹ (mg protein⁻¹). Using these kinetic values with in situ sulfide concentrations and light fluxes, we calculated that a small population of Chlorobi similar to those in enrichment CB11 can account for the observed anaerobic light-dependent sulfide consumption activity in natural water samples. We conclude that Chlorobi play a far larger role in the Chesapeake Bay than currently appreciated. This result has potential implications for coastal anoxic waters and expanding oxygen-minimum zones as they begin to impinge on the photic zone.     

Novel mechanism for conditional aerobic growth of the anaerobic bacterium Treponema denticola

Treponema denticola, a periodontal pathogen, has recently been shown to exhibit properties of a facultative anaerobic spirochete, in contrast to its previous recognition as an obligate anaerobic bacterium. In this study, the capacity and possible mechanism of T. denticola survival and growth under aerobic conditions were investigated. Factors detrimental to the growth of T. denticola ATCC 33405, such as oxygen concentration and hydrogen sulfide (H(2)S) levels as well as the enzyme activities of gamma-glutamyltransferase, cysteinylglycinase, and cystalysin associated with the cells were monitored. The results demonstrated that T. denticola grew only at deeper levels of broth (>or=3 ml in a 10-ml tube), high inoculation ratios (>or=20% of culture in medium), and short cultivation times (相似文献   

An Integrated System for the Bioremediation of Wastewater Containing Xenobiotics and Toxcic Metals

An integrated system for the biotreatment of acidic wastewaters containing both toxic metals and organics is presented. It consists of two bioprocess stages (i) an anaerobic, SRB stage (containing alkaline‐tolerant s ulfate‐ r educing b acteria) that at pH 8 (chosen to acclimatize the bacteria in the biomedium) produces high concentrations of total sulfide ions (more than 400 mg/L) which are added to the wastewater to precipitate the heavy metals out at pH 2 as metal sulfides, and (ii) an aerobic, acidophilic stage containing heterotrophic bacteria (WJB3) that degrade organic xenobiotics. The anaerobic system was comprised of a 4‐L fluidized bed bioreactor with immobilized SRB, a mixing tank, and a precipitation tank. The effluent from the bioreactor with a high concentration of sulfide ions was fed into a mixing tank where model wastewaters containing toxic metals and phenol at pH 2 were also fed at increasing loading rates until free metal ions could be detected in the precipitation tank outlet. Then the effluent from the precipitation tank outlet was fed into a 2.5‐L aerobic bioreactor in which phenol was degraded. In this research, 100 % removal efficiencies were obtained with wastewaters containing more than 400 mg/L metal ions and 900 mg/L phenol at a 6‐h HRT of the mixing tank.     

Comparative Assessment of the Aerobic and Anaerobic Microfloras of Earthworm Guts and Forest Soils

Aerobic and anaerobic microbial potentials of guts from earthworms (Lumbricus rubellus Hoffmeister and Octolasium lacteum (Oerl.)) collected from a beech forest were evaluated. On the basis of enumeration studies, microbes capable of growth under both aerobic and anaerobic conditions were more numerous in the earthworm intestine than in the beech forest soil from which the worms were obtained. The intestine of worms displayed nearly equivalent aerobic and anaerobic microbial growth potentials; in comparison, soils displayed greater aerobic than anaerobic microbial growth potentials. Hence, the ratio of microbes capable of growth under obligately anaerobic conditions to those capable of growth under aerobic conditions was higher with the worm intestine than with the soil. Process level studies corroborated these population differentials: (i) under anaerobic conditions, worm gut homogenates consumed glucose, cellobiose, or ferulate more readily than did soil homogenates; and (ii) under aerobic conditions, worm gut homogenates consumed cellobiose or oxygen more readily than did soil homogenates. Collectively, these results reinforce the general concept that the earthworm gut is not microbiologically equivalent to soil and also suggest that the earthworm gut might constitute a microhabitat enriched in microbes capable of anaerobic growth and activity.     

Microaerophilic degradation of hexahydro‐1,3,5‐trinitro‐1,3,5‐triazine (RDX) by three Rhodococcus strains

Aim: The goal of this study was to compare the degradation of hexahydro‐1,3,5‐trinitro‐1,3,5‐triazine (RDX) by three Rhodococcus strains under anaerobic, microaerophilic (<0·04 mg l^?1 dissolved oxygen) and aerobic (dissolved oxygen (DO) maintained at 8 mg l^?1) conditions. Methods and Results: Three Rhodococcus strains were incubated with no, low and ambient concentrations of oxygen in minimal media with succinate as the carbon source and RDX as the sole nitrogen source. RDX and RDX metabolite concentrations were measured over time. Under microaerophilic conditions, the bacteria degraded RDX, albeit about 60‐fold slower than under fully aerobic conditions. Only the breakdown product, 4‐nitro‐2,4‐diazabutanal (NDAB) accumulated to measurable concentrations under microaerophilic conditions. RDX degraded quickly under both aerated and static aerobic conditions (DO allowed to drop below 1 mg l^?1) with the accumulation of both NDAB and methylenedinitramine (MEDINA). No RDX degradation was observed under strict anaerobic conditions. Conclusions: The Rhodococcus strains did not degrade RDX under strict anaerobic conditions, while slow degradation was observed under microaerophilic conditions. The RDX metabolite NDAB was detected under both microaerophilic and aerobic conditions, while MEDINA was detected only under aerobic conditions. Impact and Significance of the Study: This work confirmed the production of MEDINA under aerobic conditions, which has not been previously associated with aerobic RDX degradation by these organisms. More importantly, it demonstrated that aerobic rhodococci are able to degrade RDX under a broader range of oxygen concentrations than previously reported.