Biopurification Systems Research Articles

Optimized Rhamnolipid Production by a Pseudomonas marginalis C9 Strain Isolated from a Biopurification System to Enhance Pesticide Solubilization

Biopurification systems designed for pesticide treatment are a source of diverse bacteria with environmental and biotechnological applications, including Pseudomonas marginalis C9, which has been reported as a biosurfactant-producing bacterium. The optimization of biosurfactant produced from P. marginalis C9 to enhance the solubility of a hydrophobic pesticide of environmental interest was investigated. The response surface methodology (RSM) was used to optimize the combined effect of the initial pH (5–9), agitation (100–300 rpm), and temperature (24–32 °C) on biosurfactant production. A DASbox® automated mini-bioreactor system was used to evaluate the critical factors in biosurfactant production using a full factorial design (FFD). The results showed that the optimal culture conditions using RSM were a pH of 8.5, a temperature of 25 °C, and agitation at 200 rpm. The extraction yield of the biosurfactant was 7.40 g L−1, the surface tension was reduced to 27.45 mN m−1, and the critical micelle concentration (CMC) was 48.9 mg L−1. The FFD analysis indicated that a high agitation rate (300 rpm) strongly influenced the biosurfactant activity, regardless of the inlet oxygen supply (0.5–1.5 vvm). The rhamnolipid increased the water solubility of chlorpyrifos by 11.2- and 21.7-fold at the CMC and twice the CMC, respectively.

Proteome Changes Induced by Iprodione Exposure in the Pesticide-Tolerant Pseudomonas sp. C9 Strain Isolated from a Biopurification System

Iprodione is a pesticide that belongs to the dicarboximide fungicide family. This pesticide was designed to combat various agronomical pests; however, its use has been restricted due to its environmental toxicity and risks to human health. In this study, we explored the proteomic changes in the Pseudomonas sp. C9 strain when exposed to iprodione, to gain insights into the affected metabolic pathways and enzymes involved in iprodione tolerance and biodegradation processes. As a result, we identified 1472 differentially expressed proteins in response to iprodione exposure, with 978 proteins showing significant variations. We observed that the C9 strain upregulated the expression of efflux pumps, enhancing its tolerance to iprodione and other harmful compounds. Peptidoglycan-binding proteins LysM, glutamine amidotransferase, and protein Ddl were similarly upregulated, indicating their potential role in altering and preserving bacterial cell wall structure, thereby enhancing tolerance. We also observed the presence of hydrolases and amidohydrolases, essential enzymes for iprodione biodegradation. Furthermore, the exclusive identification of ABC transporters and multidrug efflux complexes among proteins present only during iprodione exposure suggests potential counteraction against the inhibitory effects of iprodione on downregulated proteins. These findings provide new insights into iprodione tolerance and biodegradation by the Pseudomonas sp. C9 strain.

Kinetics and sorption behavior of glyphosate and tricyclazole for their efficient retention in biomixtures

The present investigation aims to study adsorption-desorption behavior of glyphosate and tricyclazole in rice straw-compost biomixtures. To enhance pesticide adsorption and performance of the bio-purification system, rice straw-compost (BM) biomixture was mixed with wheat straw biochar (WBC, 1% and 5%), and adsorption of both pesticides in control (BM) and WBCBM(1%) and WBCBM(5%) biomixtures was compared. The kinetics study suggested that the pseudo-second-order model best explained the time-dependent adsorption of both pesticides and intraparticle adsorption was not the rate-determining step. Tricyclazole was more sorbed than glyphosate in all biomixtures which can be attributed to its lower water solubility. The WBC increased the sorption of both pesticides, but the effect varied with the nature of pesticides and biochar content. The adsorption coefficient values in BM, WBCBM(1%), and WBCBM(5%) biomixtures were 26.74, 38.16, and 51.97 (glyphosate) and 38.07, 59.94, and 84.54 (tricyclazole), respectively. The adsorption data was subjected to the Freundlich, the Langmuir, and the Temkin isotherms, and among them, the Freundlich isotherm best explained pesticide adsorption behavior. Desorption results suggested that the adsorption of glyphosate was more irreversible than tricyclazole and depended upon initial pesticide concentration. This study suggested that biochar mixed rice straw-compost biomixtures can be exploited in bio-purification systems for glyphosate and tricyclazole.

Biological treatment of pesticide-containing wastewater from coffee crops: selection and optimization of a biomixture and biobed design.

The biopurification systems (BPS) or biobeds are employed for the treatment of pesticide-containing wastewater of agricultural origin. The use of these devices for pesticide removal requires the proper optimization of the composition of biomixtures (BPS active matrix) according to the target pesticides applied on a specific crop and the available materials used in their elaboration. This work aims to design a biomixture for the simultaneous treatment of several pesticides applied in coffee crops, according to local practices in Costa Rica. Three biomixtures containing either coffee husk, coconut fiber or rice husk (as the lignocellulosic substrate) were applied for the removal of 12 pesticides. The profiles of pesticide elimination and the mineralization of radiolabeled chlorpyrifos (14C-chlorpyrifos) revealed that the best performance was achieved with the coconut fiber biomixture, even though similar detoxification patterns were determined in every biomixture (according to immobilization in Daphnia magna and germination tests in Lactuca sativa). The optimization of this biomixture's composition by means of a central composite design permitted the definition of two optimal compositions (compost:soil:coconut fiber, % v/v) that maximized pesticide removal: i. 29:7.3:63.7 and ii. 11:7.3:81.7. The validation of these optimized compositions also included the use of an alternative soil from another coffee farm and resulted in overall DT50 values of 7.8-9.0 d for the pesticide mixture. Considering the removal kinetics in the optimized biomixture, a 1 m3 BPS prototype was dimensioned to be eventually used in local coffee farms. This work provides relevant information for the design and implementation of BPS at on-farm conditions for the treatment of pesticide-containing wastewater of a major crop.

Effect of the presence of carbendazim on imidacloprid degradation in a biological organic mixture

ABSTRACT The biological organic mixture (BOM) is a major component of a biopurification system (BPS), which provides an ample substrate for many microbes. Fungi present in BOM predominantly degrade pesticides through co-metabolism. We assume that the application of fungicides could have an impact on the fungal growth as well as the depuration ability of the BPS. This study is conducted to evaluate the efficiency of BOM in degrading imidacloprid in the presence of carbendazim, a commonly used fungicide. Both pesticides degraded at a faster rate when treated separately with the BOM, but their half-lives increased when applied in combination. The applied quantity of carbendazim (100 times of field recommended dose) was degraded within 60 days of the experiment. Approximately 75% of applied imidacloprid was dissipated within first 15 days of the experiment. Although carbendazim degraded at a faster rate, it had a negative impact on imidacloprid degradation during the experiment. Imidacloprid residue did not produce a negative effect on carbendazim degradation. Microbial and major soil enzyme activities were suppressed when BOM was exposed to the combination of pesticides in comparison to their single applications. Application of carbendazim was lethal to fungal population and may be the reason behind the increased persistence of imidacloprid in BOM.

Production and characterization of a biosurfactant produced by Bacillus amyloliquefaciens C11 for enhancing the solubility of pesticides

The production and characterization of lipopeptide biosurfactant obtained from a novel bacterial strain Bacillus amyloliquefaciens C11 isolated from a biopurification system, to enhance the solubility of three pesticides of environmental concern was investigated. Biosurfactant production by B. amyloliquefaciens C11 was evaluated in a 2-L reactor and characterized by chromatographic and spectroscopic methods. Biosurfactant production was associated with bacterial growth and glucose consumption. The produced biosurfactant decreased the surface tension of water from 72 to 28 mN m−1 at 24 h of incubation, with critical micelle concentration (CMC) of 51.5 mg L−1. The polymerase chain reaction for determining the presence of srfAA gene in B. amyloliquefaciens C11 was carried out. The strain exhibited the presence of srfAA gene with amplification at 201 bp, which is specific to surfactin homologues. The production of surfactin was corroborated by high performance liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF-MS) analyses. The HPLC analysis showed six isoforms of surfactin, being the isoform D found in mayor percentage (41%). Thermal gravimetric analysis (TGA) revealed the thermostability of the biosurfactant. The obtained biosurfactant maintained its properties during exposure to temperatures in range of 30–80 °C and pH values in range of 5–9. The surfactin produced by B. amyloliquefaciens C11 increased the solubility in water of chlorpyrifos, iprodione and atrazine by 7.5-fold, 1.3-fold, and 1.4-fold, respectively. Our results show that the biosurfactant produced by B. amyloliquefaciens C11 improves the bioavailability of pesticides and consequently could improves the efficiency of bioremediation processes.

Metabolic Profiling and Comparative Proteomic Insight in Respect of Amidases during Iprodione Biodegradation.

The fungicide iprodione (IPR) (3-(3,5-dichlorophenyl) N-isopropyl-2,4-dioxoimidazolidine-1-carboxamide) is a highly toxic compound. Although IPR has been restricted, it is still being applied in many places around the world, constituting an environmental risk. The biodegradation of IPR is an attractive option for reducing its residues. In this study, we isolated thirteen IPR-tolerant bacteria from a biopurification system designed to treat pesticides. A study of biodegradation using different strains was comparatively evaluated, and the best degradation rate of IPR was presented by Achromobacter sp. C1 with a half-life (T1/2) of 9 days. Based on a nano-LC-MS/MS analysis for the strains, proteins solely expressed in the IPR treatment were identified by highlighting the strain Achromobacter sp. C1, with 445 proteins primarily involved in the biosynthesis of secondary metabolites and microbial metabolism in diverse environments. Differentially expressed protein amidases were involved in six metabolic pathways. Interestingly, formamidase was inhibited while other cyclases, i.e., amidase and mandelamide hydrolase, were overexpressed, thereby minimizing the effect of IPR on the metabolism of strain C1. The dynamic changes in the protein profiles of bacteria that degrade IPR have been poorly studied; therefore, our results offer new insight into the metabolism of IPR-degrading microorganisms, with special attention paid to amidases.

On-Farm Removal of Pesticides: Current Insights on Biopurification Systems

On-Farm Removal of Pesticides: Current Insights on Biopurification Systems

Biodegradation of Iprodione and Chlorpyrifos Using an Immobilized Bacterial Consortium in a Packed-Bed Bioreactor.

This work provides the basis for implementing a continuous treatment system using a bacterial consortium for wastewater containing a pesticide mixture of iprodione (IPR) and chlorpyrifos (CHL). Two bacterial strains (Achromobacter spanius C1 and Pseudomonas rhodesiae C4) isolated from the biomixture of a biopurification system were able to efficiently remove pesticides IPR and CHL at different concentrations (10 to 100 mg L-1) from the liquid medium as individual strains and free consortium. The half-life time (T1/2) for IPR and CHL was determined for individual strains and a free bacterial consortium. However, when the free bacterial consortium was used, a lower T1/2 was obtained, especially for CHL. Based on these results, an immobilized bacterial consortium was formulated with each bacterial strain encapsulated individually in alginate beads. Then, different inoculum concentrations (5, 10, and 15% w/v) of the immobilized consortium were evaluated in batch experiments for IPR and CHL removal. The inoculum concentration of 15% w/v demonstrated the highest pesticide removal. Using this inoculum concentration, the packed-bed bioreactor with an immobilized bacterial consortium was operated in continuous mode at different flow rates (30, 60, and 90 mL h-1) at a pesticide concentration of 50 mg L-1 each. The performance in the bioreactor demonstrated that it is possible to efficiently remove a pesticide mixture of IPR and CHL in a continuous system. The metabolites 3,5-dichloroaniline (3,5-DCA) and 3,5,6-trichloro-2-pyridinol (TCP) were produced, and a slight accumulation of TCP was observed. The bioreactor was influenced by TCP accumulation but was able to recover performance quickly. Finally, after 60 days of operation, the removal efficiency was 96% for IPR and 82% for CHL. The findings of this study demonstrate that it is possible to remove IPR and CHL from pesticide-containing wastewater in a continuous system.

Biosurfactant Production by Bacillus amyloliquefaciens C11 and Streptomyces lavendulae C27 Isolated from a Biopurification System for Environmental Applications

Biosurfactant-producing bacteria can be found in contaminated environments such as biopurification systems (BPS) for pesticide treatments. A total of 18 isolates were screened to determine their ability to produce extracellular biosurfactants, using olive oil as the main carbon source. Out of the eighteen isolates, two strains (C11 and C27) were selected for biosurfactant production. The emulsification activities of the C11 and C27 strains using sunflower oil was 58.4 and 53.7%, respectively, and 46.6 and 48.0% using olive oil. Using molecular techniques and MALDI-TOF, the strains were identified as Bacillus amyloliquefaciens (C11) and Streptomyces lavendulae (C27). The submerged cultivation of the two selected strains was carried out in a 1 L stirred-tank bioreactor. The maximum biosurfactant production, indicated by the lowest surface tension measurement, was similar (46 and 45 mN/m) for both strains, independent of the fact that the biomass of the B. amyloliquefaciens C11 strain was 50% lower than the biomass of the S. lavendulae C27 strain. The partially purified biosurfactants produced by B. amyloliquefaciens C11 and S. lavendulae C27 were characterized as a lipopeptide and a glycolipid, respectively. These outcomes highlight the potential of the selected biosurfactant-producing microorganisms for improving pesticides’ bioavailability and therefore the degradational efficacy of BPS.

Actinobacteria bioaugmentation and substrate evaluation for biobeds useful for the treatment of atrazine residues in agricultural fields

Biopurification systems (BPS) or biobeds are bioprophylaxis systems to prevent pesticide point-source contamination, whose efficiency relies mostly on the pesticide removal capacity of the biomixture, the majority component of a BPS. The adaptation of the components of the biomixtures to local availabilities is a key aspect to ensure the sustainability of the system. In this work, the removal of atrazine (ATZ) was evaluated in biomixtures formulated with three sugarcane by-products as alternative lignocellulosic substrates. Based on the capacity of actinobacteria to tolerate and degrade diverse pesticides, the effect of biomixtures bioaugmentation with actinobacteria was evaluated as a strategy to enhance the depuration capacity of biobeds. Also, the effect of ATZ and/or the bioaugmentation on microbial developments and enzymatic activities were studied. The biomixtures formulated with bagasse, filter cake, or harvest residue, reached pesticide removal values of 37–41% at 28 d of incubation, with t1/2 between 37.9 ± 0.4 d and 52.3 ± 0.4 d. The bioaugmentation with Streptomyces sp. M7 accelerated the dissipation of the pesticide in the biomixtures, reducing ATZ t1/2 3-fold regarding the controls, and achieving up to 72% of ATZ removal. Atrazine did not exert a clear effect on microbial developments, although most of the microbial counts were less in the contaminated biomixtures at the end of the assay. The bioaugmentation improved the development of the microbiota in general, specially actinobacteria and fungi, regarding the non-bioaugmented systems. The inoculation with Streptomyces sp. M7 enhanced acid phosphatase activity and/or reversed a possible effect of the pesticide over this enzymatic activity.

Fast removal and detoxification of oxytetracycline, triazine and organophosphate pesticides in a biopurification system

Biopurification systems (BPS) are aimed at treating pesticide-containing wastewater. Since antibiotics employed in agriculture produce similar wastes, their treatment in BPS represents the logical disposal approach; nonetheless, antibiotics could hinder the biological removal of pesticides in BPS. This work aims at assessing the simultaneous removal of several pesticides (four organophosphates and four triazines) in a BPS biomixture during their co-disposal with the antibiotic oxytetracycline (OTC) at relevant concentrations (10 mg kg−1 and 50 mg kg−1). All pesticides were removed from the biomixture, regardless of the presence of OTC; among organophosphates, DT50 values were among the lowest reported in BPS or soil, ranging from 1.1 d to 9.31 d; only cadusafos showed a similar removal than in soil (DT50 56–61 d). Successful and fast removal was also achieved for triazines, with DT50 values ranging from 6.9 d to 11 d. Interestingly, co-application of OTC enhanced the removal of methamidophos; while in the case of triazines, OTC (at 50 mg L−1) produced a slight increase in the DT50 for every herbicide, an effect only evident for terbutryn at OTC 10 mg L−1. The system also proved to be efficient in removing OTC itself, achieving DT50 values from 8.0 to 12.3 d, the fastest so far reported in BPS. Important detoxification, over 90% towards D. magna and 26–37% on the germination index in L. sativa, provide additional support to suggest the co-disposal of OTC and the studied pesticides in the same BPS as a feasible agricultural practice at real on-farm conditions.

A novel and affordable bioaugmentation strategy with microbial extracts to accelerate the biodegradation of emerging contaminants in different media

This study describes a new bioaugmentation alternative based on the application of aqueous aerated extracts from a biomixture acclimated with ibuprofen, diclofenac and triclosan. This bioaugmentation strategy was assayed in biopurification systems (BPS) and in contaminated aqueous solutions to accelerate the removal of these emerging contaminants. Sterilized extracts or extracts from the initial uncontaminated biomixture were used as controls. In BPS, the dissipation of 90% of diclofenac and triclosan required, respectively, 60 and 108 days less than in the controls. The metabolite methyl-triclosan was determined at levels 12 times lower than in controls. In the bioaugmented solutions, ibuprofen was almost completely eliminated (99%) in 21 days and its hydroxylated metabolites were also determined to be at lower levels than in the controls. The plasmidome of acclimated biomixtures and its extract appeared to maintain certain types of plasmids but degradation related genes became less evident. Several dominant OTUs found in the extract identified as Flavobacterium and Fluviicola of the phylum Bacteroidetes, Thermomicrobia (phylum Chloroflexi) and Nonomuraea (phylum Actinobacteria), may be responsible for the enhanced dissipation of these contaminants. This bioaugmentation strategy represents an advantageous tool to facilitate in situ bioaugmentation.

Ash and biochar mixed biomixtures to degrade co-applied atrazine and fipronil in bio-augmented biobeds

ABSTRACT Retention and degradation of co-applied atrazine and fipronil was studied in laboratory constructed bio-augmented biobeds containing rice straw-compost (BM) and compost replaced with 10% rice husk ash [RHABM(10%)] or 1% wheat straw biochar [WBCBM(1%)] biomixtures. Atrazine (5000 µg, 50% WP) and fipronil (5000 µg, 0.3 G) were co-applied in 100 mL water. After 30 days, 7.31, 4.65, and 11.21 µg g−1 atrazine and 21.59, 7.41, and 37.20 µg g−1 fipronil remained undegraded in the top 3–4 cm of BM, RHABM(10%), WBCBM(1%) biobeds, respectively, suggesting that degradation of both pesticides significantly varied (p < 0.05) among the three biobeds. After 60 days 94.5%–98.3% of initially applied atrazine and 80.3%–86.9% fipronil degradation was observed. Hydroxyatrazine and sulphone were detected as the metabolites of atrazine and fipronil, respectively. Out of 5000 µg of initially applied pesticides, BM, RHABM(10%), and WBCBM(1%) biobeds accounted for 95.2%, 93.1%, and 95.2% of atrazine + hydroxyatrazine and 62.8%, 67.0%, and 71.5% of fipronil + sulphone degradation, respectively. Maximum amounts of recovered atrazine (68.8%–81.1%) and fipronil (89.8%–94.6%) were retained in the 0–5 cm sections of the biobeds. No significant effect of co-applied atrazine + fipronil was evident on the microbial parameters. This study suggested that RHA/WBC-mixed rice straw-compost biomixtures can be exploited for the detoxification of these pesticides in bio-purification systems.

Using biobeds for the treatment of fungicide-contaminated effluents from various agro-food processing industries: Microbiome responses and mobile genetic element dynamics

Agro-food processing industries generate large amounts of pesticide-contaminated effluents that pose a significant environmental threat if managed improperly. Biopurification systems like biobeds could be utilized for the depuration of these effluents although direct evidence for their efficiency are still lacking. We employed a column leaching experiment with pilot biobeds to (i) assess the depuration potential of biobeds against fungicide-contaminated effluents from seed-producing (carboxin, metalaxyl-M, fluxapyroxad), bulb-handling (thiabendazole, fludioxonil and chlorothalonil) and fruit-packaging (fludioxonil, imazalil) industries, (ii) to monitor microbial succession via amplicon sequencing and (iii) to determine the presence and dynamics of mobile genetic elements like intl1, IS1071, IncP-1 and IncP-1ε often associated with the transposition of pesticide-degrading genes. Biobeds could effectively retain (adsorbed but extractable with organic solvents) and dissipate (degraded and/or not extractable with organic solvents) the fungicides that were contained in the agro-industrial effluents with 93.1–99.98% removal efficiency in all cases. Lipophilic substances like fluxapyroxad were mostly retained in the biobed while more polar substances like metalaxyl-M and carboxin were mostly dissipated or showed higher leaching potential like metalaxyl-M. Biobeds supported a bacterial and fungal community that was not affected by fungicide application but showed clear temporal patterns in the different biobed horizons. This was most probably driven by the establishment of microaerophilic conditions upon water saturation of biobeds, as supported by the significant increase in the abundance of facultative or strict anaerobes like Chloroflexi/Anaerolinae, Acidibacter and Myxococcota. Wastewater application did not affect the dynamics of mobile genetic elements in biobeds whose abundance (intl1, IS1071, IncP-1ε) showed significant increases with time. Our findings suggest that biobeds could effectively decontaminate fungicide-contaminated effluents produced by agro-food industries and support a rather resilient microbial community.

Long-Range PCR Reveals the Genetic Cargo of IncP-1 Plasmids in the Complex Microbial Community of an On-Farm Biopurification System Treating Pesticide-Contaminated Wastewater.

Promiscuous plasmids like IncP-1 plasmids play an important role in the bacterial adaptation to pollution by acquiring and distributing xenobiotic catabolic genes. However, most information comes from isolates and the role of plasmids in governing community-wide bacterial adaptation to xenobiotics and other adaptive forces is not fully understood. Current information on the contribution of IncP-1 plasmids in community adaptation is limited because methods are lacking that directly isolate and identify the plasmid borne adaptive functions in whole-community DNA. In this study, we optimized long-range PCR to directly access and identify the cargo carried by IncP-1 plasmids in environmental DNA. The DNA between the IncP-1 backbone genes trbP and traC, a main insertion site of adaptive trait determinants, is amplified and its content analyzed by high-throughput sequencing. The method was applied to DNA of an on-farm biopurification system (BPS), treating pesticide contaminated wastewater, to examine whether horizontal gene exchange of catabolic functions by IncP-1 plasmids is a main driver of community adaptation in BPS. The cargo recovered from BPS community DNA encoded catabolic but also resistance traits and various other (un)known functions. Unexpectedly, genes with catabolic traits composed only a minor fraction of the cargo, indicating that the IncP-1 region between trbP and traC is not a major contributor to catabolic adaptation of the BPS microbiome. Instead, it contains a functionally diverse set of genes which either may assist biodegradation functions, be remnants of random gene recruitment, or confer other crucial functions for proliferation in the BPS environment. IMPORTANCE This study presents a long-range PCR for direct and cultivation-independent access to the identity of the cargo of a major insertion hot spot of adaptive genes in IncP-1 plasmids and hence a new mobilome tool for understanding the role of IncP-1 plasmids in complex communities. The method was applied to DNA of an on-farm biopurification system (BPS) treating pesticide-contaminated wastewater, aiming at new insights on whether horizontal exchange of catabolic functions by IncP-1 plasmids is a main driver of community adaptation in BPS. Unexpectedly, catabolic functions represented a small fraction of the cargo genes while multiple other gene functions were recovered. These results show that the cargo of the target insertion hot spot in IncP-1 plasmids in a community, not necessarily relates to the main obvious selective trait imposed on that community. Instead, these functions might contribute to adaptation to unknown selective forces or represent remnants of random gene recruitment.

Effectiveness of a simple biomixture for the adsorption and elimination of 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide and its metabolite, 2,4-dichlorophenol (2,4-DCP), for a biobed system

The remediation of point-source contamination of herbicides in productive fields can be addressed with biopurification systems which require biomixtures of natural materials with effectiveness for the adsorption and degradation of such compounds. A biomixture of wheat stubble and soil was tested for the adsorption and degradation of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and one of its main metabolites, 2,4-dichlorophenol (2,4-DCP). Single-contaminant kinetic studies were performed, being the pseudo-second order kinetic model the most suitable to describe the adsorption of both compounds. Although the adsorption of 2,4-D was faster, the adsorption of 2,4-DCP was three times higher. Single and competitive adsorption isotherms were also obtained. The later were acquired from a complete factorial experimental design varying the concentration of both solutes. Mathematical models were applied to describe the single solute isotherms, given a maximum monolayer adsorption capacity of 155.7 mg kg−1 for 2,4-D and 371.6 mg kg−1 for 2,4-DCP. Multicomponent models for bi-solute isotherms showed a clear reduction of 2,4-D adsorption in the presence of 2,4-DCP, although no adsorption displacement for the 2,4 DCP metabolite was observed. A stagnant degradation study was run and the contaminants were surveyed during 30 days. The biomixture was successful in degrading the 2,4-D, with a removal effectiveness > 96% after 15 days. The 2,4-DCP was not detected, suggesting a very fast degradation as soon as generated.

BioF is a novel B2 metallo-β-lactamase from Pseudomonas sp. isolated from an on-farm biopurification system.

Antimicrobial resistance represents a major global health concern and environmental bacteria are considered a source of resistance genes. Carbapenems are often used as the last antibiotic option to treat multidrug-resistant bacteria. Metallo-β-lactamases (MBLs) are able to render resistance to almost all β-lactam antibiotics, including carbapenems. Unfortunately, there are no inhibitors against MBLs for clinical use. Subclass B2 MBLs are the only enzymes working as strict carbapenemases, under-represented, encoded in chromosome genes and only functional as mono-zinc enzymes. Despite current efforts in MBLs inhibitor development, B2 carbapenemase activity is especially difficult to suppress, even in vitro. In this study we characterized BioF, a novel subclass B2 MBL identified in a new environmental Pseudomonas sp. strain isolated from an on-farm biopurification system (BPS). Although blaBioF is most likely a chromosomal gene, it is found in a genomic island and may represent a step previous to the horizontal transmission of B2 genes. The new B2 MBL is active as a mono-zinc enzyme and is a potent carbapenemase with incipient activity against some cephalosporins. BioF activity is not affected by excess zinc and is only inhibited at high metal chelator concentrations. The discovery and characterization of B2 MBL BioF as a potent carbapenemase in a BPS bacterial isolate emphasizes the importance of exploring antibiotic resistances existing in the environmental microbiota under the influence of human activities before they could emerge clinically.

Evaluating ash and biochar mixed biomixtures for atrazine and fipronil degradation

Composition of biomixtures is crucial for the success of the bio-purification system. Therefore, compost in a rice straw-compost biomixture (BM) was partially replaced with rice husk ash (RHA, 5 & 10%), sugarcane bagasse ash (SBA, 5 & 10%) and wheat straw biochar (WBC, 1 & 5%). Degradation of atrazine and fipronil was studied in these biomixtures and effects of pesticide concentration (50 and 100 μ g g −1 ) and bio-augmentation with pesticide-degrading microbial cultures were evaluated. Ashes and biochar had varied effects on atrazine and fipronil degradation that followed first order kinetics, but, was biphasic for atrazine degradation. Atrazine degradation during the II nd phase was faster than the I st phase. Compared to control (BM), RHA decreased the half-life ( t 1∕2 ) of atrazine at lower concentration (50 μ g g −1 ) while no significant effect of SBA and WBC (1%) was observed. However, WBC (5%) increased atrazine t 1∕2 by 200% (50 μ g g −1 ) and 300% (100 μ g g −1 ). No significant effect of RHA, SBA and WBC (1%) was evident on fipronil degradation and the t 1∕2 varied between 11.63–15.42 days, while, WBC (5%) slightly increased the t 1∕2 by 35%–40%. These results suggested that atrazine degradation in WBC-mixed biomixtures was limited by its availability due to higher adsorption, while higher fipronil adsorption in WBC-mixed biomixtures did not affect its degradation. Bio-augmentation of biomixtures with pesticide degrading cultures increased degradation of atrazine and fipronil and their metabolites, hydroxyatrazine and fipronil sulfone, respectively and the effect was significant on atrazine/hydroxyatrazine degradation. No significant effect of ashes/biochar was evident on microbial biomass carbon, dehydrogenase and fluorescein diacetate activity. This study suggested that bio-augmentation of ashes/biochar-mixed rice straw-compost biomixtures can be exploited for detoxification of these pesticides in bio-purification systems. • Rice husk and sugarcane bagasse ashes (10%) mixed biomixtures did not inhibit atrazine concentration. • Wheat straw biochar at 5% levels inhibited degradation of atrazine. • No effect of amendments was evident on fipronil degradation. • Bio-augmentation significantly increased atrazine degradation.

Straw-Based Biopurification Systems to Remove Ibuprofen, Diclofenac and Triclosan from Wastewaters: Dominant Microbial Communities

The continued discharge of pharmaceuticals and personal care products (PPCPs) into the environment due to their widespread use and the lack of effective systems for their removal from water is a global problem. In this study, the dissipation of ibuprofen, diclofenac and triclosan added simultaneously in biopurification systems (BPSs) with different compositions and their effect on the microbial community structure was analysed. Three BPSs, constituted by mixtures of soil (S), peat (P), or raw wet olive mill cake (A) or its vermicompost (V) and straw (S) were prepared (SPS, SAS and SVS). Sorption and degradation experiments were carried out. After 84 days of incubation, more than 85% of each PPCP applied had dissipated. Methyl-triclosan was determined to be highest in the SVS biomixture. Biomixtures with lower C/N ratio and higher alpha diversity were the most effective in the removal of PPCPs. Initially, the BPS biomixtures showed a different microbial structure dominated by Proteobacteria, Actinobacteria and Bacteroidetes but after addition of PPCPs, a similar pattern was observed in the relative abundance of the phylum Chloroflexi, the class Sphingobacteriia and the genus Brevundimonas. These biopurification systems can be useful to prevent point source contamination due to the disposal of PPCP-contaminated waters.