The Growth Yield of Aminobacter niigataensis MSH1 on the Micropollutant 2,6-Dichlorobenzamide Decreases Substantially at Trace Substrate Concentrations.

2,6-Dichlorobenzamide (BAM) is an omnipresent micropollutant in European groundwaters. Aminobacter niigataensis MSH1 is a prime candidate for biologically treating BAM-contaminated groundwater since this organism is capable of utilizing BAM as a carbon and energy source. However, detailed information on the BAM degradation kinetics by MSH1 at trace concentrations is lacking, while this knowledge is required for predicting and optimizing the degradation process. Contaminating assimilable organic carbon (AOC) in media makes the biodegradation experiment a mixed-substrate assay and hampers exploration of pollutant degradation at trace concentrations. In this study, we examined how the BAM concentration affects MSH1 growth and BAM substrate utilization kinetics in a AOC-restricted background to avoid mixed-substrate conditions. Conventional Monod kinetic models were unable to predict kinetic parameters at low concentrations from kinetics determined at high concentrations. Growth yields on BAM were concentration-dependent and decreased substantially at trace concentrations; i.e., growth of MSH1 diminished until undetectable levels at BAM concentrations below 217 µg-C/L. Nevertheless, BAM degradation continued. Decreasing growth yields at lower BAM concentrations might relate to physiological adaptations to low substrate availability or decreased expression of downstream steps of the BAM catabolic pathway beyond 2,6-dichlorobenzoic acid (2,6-DCBA) that ultimately leads to Krebs cycle intermediates for growth and energy conservation.

Interspecies Interactions of the 2,6-Dichlorobenzamide Degrading Aminobacter sp. MSH1 with Resident Sand Filter Bacteria: Indications for Mutual Cooperative Interactions That Improve BAM Mineralization Activity.

Bioaugmentation often involves an invasion process requiring the establishment and activity of a foreign microbe in the resident community of the target environment. Interactions with resident micro-organisms, either antagonistic or cooperative, are believed to impact invasion. However, few studies have examined the variability of interactions between an invader and resident species of its target environment, and none of them considered a bioremediation context. Aminobacter sp. MSH1 mineralizing the groundwater micropollutant 2,6-dichlorobenzamide (BAM), is proposed for bioaugmentation of sand filters used in drinking water production to avert BAM contamination. We examined the nature of the interactions between MSH1 and 13 sand filter resident bacteria in dual and triple species assemblies in sand microcosms. The residents affected MSH1-mediated BAM mineralization without always impacting MSH1 cell densities, indicating effects on cell physiology rather than on cell number. Exploitative competition explained most of the effects (70%), but indications of interference competition were also found. Two residents improved BAM mineralization in dual species assemblies, apparently in a mutual cooperation, and overruled negative effects by others in triple species systems. The results suggest that sand filter communities contain species that increase MSH1 fitness. This opens doors for assisting bioaugmentation through co-inoculation with "helper" bacteria originating from and adapted to the target environment.

The complete genome of 2,6-dichlorobenzamide (BAM) degrader Aminobacter sp. MSH1 suggests a polyploid chromosome, phylogenetic reassignment, and functions of plasmids.

Aminobacter sp. MSH1 (CIP 110285) can use the pesticide dichlobenil and its recalcitrant transformation product, 2,6-dichlorobenzamide (BAM), as sole source of carbon, nitrogen, and energy. The concentration of BAM in groundwater often exceeds the threshold limit for drinking water, requiring additional treatment in drinking water treatment plants or closure of the affected abstraction wells. Biological treatment with MSH1 is considered a potential sustainable alternative to remediate BAM-contamination in drinking water production. We present the complete genome of MSH1, which was determined independently in two institutes at Aarhus University and KU Leuven. Divergences were observed between the two genomes, i.e. one of them lacked four plasmids compared to the other. Besides the circular chromosome and the two previously described plasmids involved in BAM catabolism, pBAM1 and pBAM2, the genome of MSH1 contained two megaplasmids and three smaller plasmids. The MSH1 substrain from KU Leuven showed a reduced genome lacking a megaplasmid and three smaller plasmids and was designated substrain MK1, whereas the Aarhus variant with all plasmids was designated substrain DK1. A plasmid stability experiment indicate that substrain DK1 may have a polyploid chromosome when growing in R2B medium with more chromosomes than plasmids per cell. Finally, strain MSH1 is reassigned as Aminobacter niigataensis MSH1.

Bacteriophage-mediated interference of the c-di-GMP signalling pathway in Pseudomonas aeruginosa.

C-di-GMP is a key signalling molecule which impacts bacterial motility and biofilm formation and is formed by the condensation of two GTP molecules by a diguanylate cyclase. We here describe the identification and characterization of a family of bacteriophage-encoded peptides that directly impact c-di-GMP signalling in Pseudomonas aeruginosa. These phage proteins target Pseudomonas diguanylate cyclase YfiN by direct protein interaction (termed YIPs, YfiN Interacting Peptides). YIPs induce an increase of c-di-GMP production in the host cell, resulting in a decrease in motility and an increase in biofilm mass in P. aeruginosa. A dynamic analysis of the biofilm morphology indicates a denser biofilm structure after induction of the phage protein. This intracellular signalling interference strategy by a lytic phage constitutes an unexplored phage-based mechanism of metabolic regulation and could potentially serve as inspiration for the development of molecules that interfere with biofilm formation in P. aeruginosa and other pathogens.

Detection of Lipid Oxidation in Infant Formulas: Application of Infrared Spectroscopy to Complex Food Systems.

 Fish- or algal oils have become a common component of infant formula products for their high docosahexaenoic acid (DHA) content. DHA is widely recognized to contribute to the normal development of the infant, and the European Commission recently regulated the DHA content in infant formulas. For many manufacturers of first-age early life nutrition products, a higher inclusion level of DHA poses various challenges. Long-chain polyunsaturated fatty acids (LC-PUFAs) such as DHA are very prone to oxidation, which can alter the organoleptic property and nutritional value of the final product. Traditional methods for the assessment of oxidation in complex systems require solvent extraction of the included fat, which can involve harmful reagents and may alter the oxidation status of the system. A rapid, efficient, non-toxic real-time method to monitor lipid oxidation in complex systems such as infant formula emulsions would be desirable. In this study, infrared spectroscopy was therefore chosen to monitor iron-induced oxidation in liquid infant formula, with conjugated dienes and headspace volatiles measured with GC-MS as reference methods. Infrared spectra of infant formula were recorded directly in mid- and near-infrared regions using attenuated total reflectance Fourier-transform (ATR-FTIR) and near-infrared (NIRS) spectrophotometers. Overall, good correlation coefficients (R2 > 0.9) were acquired between volatiles content and infrared spectroscopy. Despite the complex composition of infant formula containing proteins and sugars, infrared spectroscopy was still able to detect spectral changes unique to lipid oxidation. By comparison, near-infrared spectroscopy (NIRS) presented better results than ATR-FTIR: prediction error ATR-FTIR 18% > prediction error NIRS 9%. Consequently, NIRS demonstrates great potential to be adopted as an in-line or on-line, non-destructive, and sustainable method for dairy and especially infant formula manufacturers.

Aminobacter sp. MSH1 Mineralizes the Groundwater Micropollutant 2,6-Dichlorobenzamide through a Unique Chlorobenzoate Catabolic Pathway.

2,6-Dichlorobenzamide (BAM) is a major groundwater micropollutant posing problems for drinking water treatment plants (DWTPs) that depend on groundwater intake. Aminobacter sp. MSH1 uses BAM as the sole source of carbon, nitrogen, and energy and is considered a prime biocatalyst for groundwater bioremediation in DWTPs. Its use in bioremediation requires knowledge of its BAM-catabolic pathway, which is currently restricted to the amidase BbdA converting BAM into 2,6-dichlorobenzoic acid (2,6-DCBA) and the monooxygenase BbdD transforming 2,6-DCBA into 2,6-dichloro-3-hydroxybenzoic acid. Here, we show that the 2,6-DCBA catabolic pathway is unique and differs substantially from catabolism of other chlorobenzoates. BbdD catalyzes a second hydroxylation, forming 2,6-dichloro-3,5-dihydroxybenzoic acid. Subsequently, glutathione-dependent dehalogenases (BbdI and BbdE) catalyze the thiolytic removal of the first chlorine. The remaining chlorine is then removed hydrolytically by a dehalogenase of the α/ß hydrolase superfamily (BbdC). BbdC is the first enzyme in that superfamily associated with dehalogenation of chlorinated aromatics and appears to represent a new subtype within the α/ß hydrolase dehalogenases. The activity of BbdC yields a unique trihydroxylated aromatic intermediate for ring cleavage that is performed by an extradiol dioxygenase (BbdF) producing 2,4,6-trioxoheptanedioic acid, which is likely converted to Krebs cycle intermediates by BbdG.

The pesticide mineralization capacity in sand filter units of drinking water treatment plants (DWTP): Consistency in time and relationship with intake water and sand filter characteristics.

Sand filters (SFs) are commonly applied in drinking water treatment plants (DWTPs) for removal of iron and manganese but also show potential for microbial degradation of pesticide residues. The latter is advantageous in case the intake water contains pesticide residues. However, whether this involves mineralization suggesting no generation of harmful transformation products, its consistency over time, and how this ability relates to physicochemical and biological characteristics of the DWTP intake water and the SFs is unknown. The capacity to mineralize the herbicides bentazon and 2-methyl-4-chlorophenoxyacetic acid (MCPA) was examined in SF samples from 11 DWTPs differing in operation, intake water composition and pesticide contamination level. MCPA was mineralized in all biologically active SFs while mineralization of bentazon occurred rarely. Mineralization of both compounds was consistent in time and across samples taken from different SF units of the same DWTP. Kinetic modelling of mineralization curves suggested the occurrence of growth linked bentazon and MCPA mineralization in several SF samples. Multivariate analysis correlating intake water/SF characteristics with pesticide mineralization indicated that pesticide mineralization capacity depended on a range of intake water characteristics, but was not necessarily explained by the presence of the pesticide in the intake water and hence the in situ exposure of the SF community to the pesticide. This was supported by testing a sample from DWTP Kluizen for its capacity to mineralize 5 other pesticides including pesticides not present or occasionally present in the intake water. All of those pesticides were mineralized as well.

Intra- and inter-field diversity of 2,4-dichlorophenoxyacetic acid-degradative plasmids and their tfd catabolic genes in rice fields of the Mekong delta in Vietnam.

The tfd genes mediating degradation of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) differ in composition and organization in bacterial isolates from different geographical origin and are carried by different types of mobile genetic elements (MGE). It is not known whether such global diversity of 2,4-D-catabolic MGE and their tfd gene cargo is reflected in the diversity at field scale. The genomic context of the 2,4-D catabolic genes of 2,4-D-degrading isolates from two rice fields with a 2,4-D application history, located in two distant provinces of the Vietnam Mekong delta, was compared. All isolates were ß-proteobacteria, were unique for each rice field and carried the catabolic genes on MGE and especially plasmids. Most plasmids were IncP-1ß plasmids and carried tfd clusters highly similar to those of the IncP-1ß plasmid pJP4, typified by two chlorophenol catabolic gene modules (tfd-I and tfd-II). IncP-1ß plasmids from the same field showed small deletions and/or insertions in accessory metabolic genes. One plasmid belonged to an unclassified plasmid group and carries a copy of both tfdA and tfd-II identical to those in the IncP-1ß plasmids. Our results indicate intra-field evolution and inter-field exchange of 2,4-D-catabolic IncP-1ß plasmids as well as the exchange of tfd genes between different plasmids within a confined local environment.

Catabolism of the groundwater micropollutant 2,6-dichlorobenzamide beyond 2,6-dichlorobenzoate is plasmid encoded in Aminobacter sp. MSH1.

Aminobacter sp. MSH1 uses the groundwater micropollutant 2,6-dichlorobenzamide (BAM) as sole source of carbon and energy. In the first step, MSH1 converts BAM to 2,6-dichlorobenzoic acid (2,6-DCBA) by means of the BbdA amidase encoded on the IncP-1ß plasmid pBAM1. Information about the genes and degradation steps involved in 2,6-DCBA metabolism in MSH1 or any other organism is currently lacking. Here, we show that the genes for 2,6-DCBA degradation in strain MSH1 reside on a second catabolic plasmid in MSH1, designated as pBAM2. The complete sequence of pBAM2 was determined revealing that it is a 53.9 kb repABC family plasmid. The 2,6-DCBA catabolic genes on pBAM2 are organized in two main clusters bordered by IS elements and integrase genes and encode putative functions like Rieske mono-/dioxygenase, meta-cleavage dioxygenase, and reductive dehalogenases. The putative mono-oxygenase encoded by the bbdD gene was shown to convert 2,6-DCBA to 3-hydroxy-2,6-dichlorobenzoate (3-OH-2,6-DCBA). 3-OH-DCBA was degraded by wild-type MSH1 and not by a pBAM2-free MSH1 variant indicating that it is a likely intermediate in the pBAM2-encoded DCBA catabolic pathway. Based on the activity of BbdD and the putative functions of the other catabolic genes on pBAM2, a metabolic pathway for BAM/2,6-DCBA in strain MSH1 was suggested.

Catabolic task division between two near-isogenic subpopulations co-existing in a herbicide-degrading bacterial consortium: consequences for the interspecies consortium metabolic model.

Variovorax sp. WDL1 mediates hydrolysis of the herbicide linuron into 3,4-dichloroaniline (DCA) and N,O-dimethylhydroxylamine in a tripartite bacterial consortium with Comamonas testosteroni WDL7 and Hyphomicrobium sulfonivorans WDL6. Although strain WDL1 contains the dcaQTA1A2B operon for DCA oxidation, this conversion is mainly performed by WDL7. Phenotypic diversification observed in WDL1 cultures and scrutiny of the WDL1 genome suggest that WDL1 cultures consist of two dedicated subpopulations, i.e., a linuron-hydrolysing subpopulation (Lin + DCA-) and a DCA-oxidizing subpopulation (Lin-DCA+). Whole genome analysis of strains representing the respective subpopulations revealed that they are identical, aside from the presence of hylA (in Lin + DCA- cells) and the dcaQTA1A2B gene cluster (in Lin-DCA+ cells), and that these catabolic gene modules replace each other at exactly the same locus on a 1380 kb extra-chromosomal element that shows plasmid gene functions including genes for transferability by conjugation. Both subpopulations proliferate in consortium biofilms fed with linuron, but Lin + DCA- cells compose the main WDL1 subpopulation. Our observations instigated revisiting the interactions within the consortium and suggest that the physical separation of two essential linuron catabolic gene clusters in WDL1 by mutually exclusive integration in the same mobile genetic element is key to the existence of WDL1 in a consortium mode.

Groundwater contamination with 2,6-dichlorobenzamide (BAM) and perspectives for its microbial removal.

The pesticide metabolite 2,6-dichlorobenzamide (BAM) is very persistent in both soil and groundwater and has become one of the most frequently detected groundwater micropollutants. BAM is not removed by the physico-chemical treatment techniques currently used in drinking water treatment plants (DWTP); therefore, if concentrations exceed the legal threshold limit, it represents a sizeable problem for the stability and quality of drinking water production, especially in places that depend on groundwater for drinking water. Bioremediation is suggested as a valuable strategy for removing BAM from groundwater by deploying dedicated BAM-degrading bacteria in DWTP sand filters. Only a few bacterial strains with the capability to degrade BAM have been isolated, and of these, only three isolates belonging to the Aminobacter genus are able to mineralise BAM. Considerable effort has been made to elucidate degradation pathways, kinetics and degrader genes, and research has recently been presented on the application of strain Aminobacter sp. MSH1 for the purification of BAM-contaminated water. The aim of the present review was to provide insight into the issue of BAM contamination and to report on the current status and knowledge with regard to the application of microorganisms for purification of BAM-contaminated water resources. This paper discusses the prospects and challenges for bioaugmentation of DWTP sand filters with specific BAM-degrading bacteria and identifies relevant perspectives for future research.

Aminobacter sp. MSH1 invades sand filter community biofilms while retaining 2,6-dichlorobenzamide degradation functionality under C- and N-limiting conditions.

Aminobacter sp. MSH1 is of interest for bioaugmentation of biofiltration units in drinking water treatment plants (DWTPs) due to its ability to degrade the groundwater micropollutant 2,6-dichlorobenzamide (BAM). Using a continuous flow chamber biofilm model, MSH1 was previously shown to colonize surfaces and degrade BAM at trace concentrations as low as 1 µg/L under the oligotrophic conditions found in DWTPs. In DWTP filtration units, MSH1 has to compete with the resident biofilm microbiota for space and nutrients. Using the same model, we examined how a sand filter community (SFC) affects MSH1's BAM-degrading activity and biofilm formation under C- and N-limiting conditions when fed with trace concentrations of BAM. MSH1 was inoculated simultaneously with the SFC (co-colonization mode) or after the SFC formed a biofilm (invasion mode). MSH1 successfully established in the SFC biofilm showing growth and activity. In co-colonization mode, MSH1 decreased in number in the presence of the SFC and formed isolated colonies, while specific BAM-degradation activity increased. In the invasion mode, MSH1 also decreased in numbers in the presence of the SFC but formed mixed colonies, while specific BAM degradation was unaffected. Our results show that MSH1 invades and performs successfully in an SFC biofilm under the oligotrophic conditions of DWTPs.

Genetic (In)stability of 2,6-Dichlorobenzamide Catabolism in Aminobacter sp. Strain MSH1 Biofilms under Carbon Starvation Conditions.

Aminobacter sp. strain MSH1 grows on and mineralizes the groundwater micropollutant 2,6-dichlorobenzamide (BAM) and is of interest for BAM removal in drinking water treatment plants (DWTPs). The BAM-catabolic genes in MSH1 are located on plasmid pBAM1, carrying bbdA, which encodes the conversion of BAM to 2,6-dichlorobenzoic acid (2,6-DCBA) (BbdA+ phenotype), and plasmid pBAM2, carrying gene clusters encoding the conversion of 2,6-DCBA to tricarboxylic acid (TCA) cycle intermediates (Dcba+ phenotype). There are indications that MSH1 easily loses its BAM-catabolic phenotype. We obtained evidence that MSH1 rapidly develops a population that lacks the ability to mineralize BAM when grown on nonselective (R2B medium) and semiselective (R2B medium with BAM) media. Lack of mineralization was explained by loss of the Dcba+ phenotype and corresponding genes. The ecological significance of this instability for the use of MSH1 for BAM removal in the oligotrophic environment of DWTPs was explored in lab and pilot systems. A higher incidence of BbdA+ Dcba- MSH1 cells was also observed when MSH1 was grown as a biofilm in flow chambers under C and N starvation conditions due to growth on nonselective residual assimilable organic carbon. Similar observations were made in experiments with a pilot sand filter reactor bioaugmented with MSH1. BAM conversion to 2,6-DCBA was not affected by loss of the DCBA-catabolic genes. Our results show that MSH1 is prone to BAM-catabolic instability under the conditions occurring in a DWTP. While conversion of BAM to 2,6-DCBA remains unaffected, BAM mineralization activity is at risk, and monitoring of metabolites is warranted.IMPORTANCE Bioaugmentation of dedicated biofiltration units with bacterial strains that grow on and mineralize micropollutants was suggested as an alternative for treating micropollutant-contaminated water in drinking water treatment plants (DWTPs). Organic-pollutant-catabolic genes in bacteria are often easily lost, especially under nonselective conditions, which affects the bioaugmentation success. In this study, we provide evidence that Aminobacter sp. strain MSH1, which uses the common groundwater micropollutant 2,6-dichlorobenzamide (BAM) as a C source, shows a high frequency of loss of its BAM-mineralizing phenotype due to the loss of genes that convert 2,6-DCBA to Krebs cycle intermediates when nonselective conditions occur. Moreover, we show that catabolic-gene loss also occurs in the oligotrophic environment of DWTPs, where growth of MSH1 depends mainly on the high fluxes of low concentrations of assimilable organic carbon, and hence show the ecological relevance of catabolic instability for using strain MSH1 for BAM removal in DWTPs.

Carbon catabolite repression and cell dispersal affect degradation of the xenobiotic compound 3,4-dichloroaniline in Comamonas testosteroni WDL7 biofilms.

Organic pollutant degrading biofilms in natural ecosystems and water treatment systems are often exposed to other carbon sources in addition to the pollutant. The availability of auxiliary carbon sources can lead to surplus biomass growth, changes in biofilm structure and carbon catabolite repression (CCR) which together will affect pollutant degradation rate and efficiency of the system. To understand the interplay between these processes, continuous biofilms of the 3,4-dichloroaniline (3,4-DCA) degrading Comamonas testosteroni WDL7-RFP were grown in single- and dual-substrate conditions with 3,4-DCA and/or citrate and reciprocal effects on 3,4-DCA/citrate degradation, biofilm biomass and biofilm structure were examined. The main mechanism affecting 3,4-DCA degradation in biofilms in dual-substrate conditions was citrate-mediated CCR as reflected by a decrease in specific 3,4-DCA degrading activity. Growth on citrate partially compensated for the lowered specific 3,4-DCA degradation activity under dual substrate conditions but not to the extent expected from growth observed under single-substrate conditions with citrate. This was explained by higher residual 3,4-DCA concentrations in the presence of citrate that increased cell dispersal in the biofilms. Our results show hampered pollutant removal in biofilms due to a complex interplay of auxiliary organic C source utilization for growth affecting the specific pollutant degradation rate and changes in cell physiology due to increased exposure to the pollutant as a result of lowered pollutant degradation rates.

Uneven distribution of inorganic pollutants in marine air originating from ocean-going ships.

The distribution of mass, water-soluble inorganic salts and mineral elements of size-segregated aerosols (PM1, PM2.5-1 and PM10-2.5), precursor gaseous pollutants, black carbon, and nanoparticles (10-300 nm size range) at the Southern Bight of the North Sea has been studied. The concentrations of air pollutants peaked over shipping lanes, open-water anchorage areas and frequently navigated waters, due to the presence of mobile emission sources. A considerable decrease in air pollutant levels was seen when diverting from these marine areas towards remote or coastal banks. These findings showed the rapid dispersion of pollutants in the marine air. The nano-aerosol count, originating from ocean-going ships, peaked at lower average aerodynamic diameters (e.g., ≈28 nm) than those, observed from low-displacement vessels (45-50 nm, e.g., for fishing boats). The average diameter of nano-PM depended also on weather conditions, e.g., it was higher (≈50 nm) in air of higher humidity.

Soil-Bacterium Compatibility Model as a Decision-Making Tool for Soil Bioremediation.

Bioremediation of organic pollutant contaminated soil involving bioaugmentation with dedicated bacteria specialized in degrading the pollutant is suggested as a green and economically sound alternative to physico-chemical treatment. However, intrinsic soil characteristics impact the success of bioaugmentation. The feasibility of using partial least-squares regression (PLSR) to predict the success of bioaugmentation in contaminated soil based on the intrinsic physico-chemical soil characteristics and, hence, to improve the success of bioaugmentation, was examined. As a proof of principle, PLSR was used to build soil-bacterium compatibility models to predict the bioaugmentation success of the phenanthrene-degrading Novosphingobium sp. LH128. The survival and biodegradation activity of strain LH128 were measured in 20 soils and correlated with the soil characteristics. PLSR was able to predict the strain's survival using 12 variables or less while the PAH-degrading activity of strain LH128 in soils that show survival was predicted using 9 variables. A three-step approach using the developed soil-bacterium compatibility models is proposed as a decision making tool and first estimation to select compatible soils and organisms and increase the chance of success of bioaugmentation.

Biocarriers Improve Bioaugmentation Efficiency of a Rapid Sand Filter for the Treatment of 2,6-Dichlorobenzamide-Contaminated Drinking Water.

Aminobacter sp. MSH1 immobilized in an alginate matrix in porous stones was tested in a pilot system as an alternative inoculation strategy to the use of free suspended cells for biological removal of micropollutant concentrations of 2,6-dichlorobenzamide (BAM) in drinking water treatment plants (DWTPs). BAM removal rates and MSH1 cell numbers were recorded during operation and assessed with specific BAM degradation rates obtained in lab conditions using either freshly grown cells or starved cells to explain reactor performance. Both reactors inoculated with either suspended or immobilized cells showed immediate BAM removal under the threshold of 0.1 µg/L, but the duration of sufficient BAM removal was 2-fold (44 days) longer for immobilized cells. The longer sufficient BAM removal in case of immobilized cells compared to suspended cells was mainly explained by a lower initial loss of MSH1 cells at operational start due to volume replacement and shear. Overall loss of activity in the reactors though was due to starvation, and final removal rates did not differ between reactors inoculated with immobilized and suspended cells. Management of assimilable organic carbon, in addition to cell immobilization, appears crucial for guaranteeing long-term BAM degradation activity of MSH1 in DWTP units.

Surface Colonization and Activity of the 2,6-Dichlorobenzamide (BAM) Degrading Aminobacter sp. Strain MSH1 at Macro- and Micropollutant BAM Concentrations.

Aminobacter sp. MSH1 uses the groundwater micropollutant 2,6-dichlorobenzamide (BAM) as a C and N source and is a potential catalyst for biotreatment of BAM-contaminated groundwater in filtration units of drinking water treatment plants (DWTPs). The oligotrophic environment of DWTPs including trace pollutant concentrations, and the high flow rates impose challenges for micropollutant biodegradation in DWTPs. To understand how trace BAM concentrations affect MSH1 surface colonization and BAM degrading activity, MSH1 was cultivated in flow channels fed continuously with BAM macro- and microconcentrations in a N- and C-limiting medium. At all BAM concentrations, MSH1 colonized the flow channel. BAM degradation efficiencies were concentration-dependent, ranging between 70 and 95%. Similarly, BAM concentration affected surface colonization, but at 100 µg/L BAM and lower, colonization was similar to that in systems without BAM, suggesting that assimilable organic carbon and nitrogen other than those supplied by BAM sustained colonization at BAM microconcentrations. Comparison of specific BAM degradation rates in flow channels and in cultures of suspended freshly grown cells indicated that starvation conditions in flow channels receiving BAM microconcentrations resulted into MSH1 biomasses with 10-100-times reduced BAM degrading activity and provided a kinetic model for predicting BAM degradation under continuous C and N starvation.

Mineralization of the Common Groundwater Pollutant 2,6-Dichlorobenzamide (BAM) and its Metabolite 2,6-Dichlorobenzoic Acid (2,6-DCBA) in Sand Filter Units of Drinking Water Treatment Plants.

The intrinsic capacity to mineralize the groundwater pollutant 2,6-dichlorobenzamide (BAM) and its metabolite 2,6-dichlorobenzoic acid (2,6-DCBA) was evaluated in samples from sand filters (SFs) of drinking water treatment plants (DWTPs). Whereas BAM mineralization occurred rarely and only in SFs exposed to BAM, 2,6-DCBA mineralization was common in SFs, including those treating uncontaminated water. Nevertheless, SFs treating BAM contaminated water showed the highest 2,6-DCBA mineralization rates. For comparison, 2,6-DCBA and BAM mineralization were determined in various topsoil samples. As in SF samples, BAM mineralization was rare, whereas 2,6-DCBA mineralization capacity appeared widespread, with high mineralization rates found especially in forest soils. Multivariate analysis showed that in both SF and soil samples, high 2,6-DCBA mineralization correlated with high organic carbon content. Adding a 2,6-DCBA degradation deficient mutant of the BAM mineralizing Aminobacter sp. MSH1 confirmed that 2,6-DCBA produced from BAM is rapidly mineralized by the endogenous microbial community in SFs showing intrinsic 2,6-DCBA mineralization. This study demonstrates that (i) 2,6-DCBA mineralization is widely established in SFs of DWTPs, allowing the mineralization of 2,6-DCBA produced during BAM degradation and (ii) the first metabolic step in BAM mineralization is rare in microbial communities, rather than its further degradation beyond 2,6-DCBA.

Application of biodegradation in mitigating and remediating pesticide contamination of freshwater resources: state of the art and challenges for optimization.

In recent years, the application of pesticide biodegradation in remediation of pesticide-contaminated matrices moved from remediating bulk soil to remediating and mitigating pesticide pollution of groundwater and surface water bodies. Specialized pesticide-degrading microbial populations are used, which can be endogenous to the ecosystem of interest or introduced by means of bioaugmentation. It involves (semi-)natural ecosystems like agricultural fields, vegetated filter strips, and riparian wetlands and man-made ecosystems like on-farm biopurification systems, groundwater treatment systems, and dedicated modules in drinking water treatment. Those ecosystems and applications impose challenges which are often different from those associated with bulk soil remediation. These include high or extreme low pesticide concentrations, mixed contamination, the presence of alternative carbon sources, specific hydraulic conditions, and spatial and temporal variation. Moreover, for various indicated ecosystems, limited knowledge exists about the microbiota present and their physiology and about the in situ degradation kinetics. This review reports on the current knowledge on applications of biodegradation in mitigating and remediating freshwater pesticide contamination. Attention is paid to the challenges involved and current knowledge gaps for improving those applications.