Biobed System Research Articles

Using the Pesticide Toxicity Index to show the potential ecosystem benefits of on-farm biobeds.

The influent and effluent of two single-cell biobeds (Province of Alberta, Canada) and two dual cell-biobeds (Province of Saskatchewan, Canada) were monitored during a number of growing seasons. A total of 59 unique pesticide active ingredients were detected, with all biobed influent samples (n=54) and 93% of effluent samples (n=54) containing pesticide mixtures. About one-half of the effluent samples in both single-cell (56%) and dual-cell (45%) biobeds contained active ingredients that have Groundwater Ubiquity Score (GUS) values >2.8 and so were more likely to move through the biomatrix materials into effluent. The Pesticide Toxicity Index (PTI) calculated for aquatic indicator species (i.e., vascular and nonvascular plants, invertebrates, and fish) was always larger for influent samples (e.g., median PTI>500 for invertebrates in dual-cell biobed) than effluent samples (i.e., median PTI<1). As such, this study demonstrates the potential ecosystem benefits of the broad adoption of on-farm biobeds in the Canadian Prairies for recycling tank rinsate as a strategy to accelerate a green economy. Although biobeds were highly effective in reducing the concentrations for pesticides with a wide range of soil organic carbon coefficient and half-life values, the biobed effectiveness was relatively poor for the herbicides clopyralid, diclofop, fluroxypyr, and imazethapyr. Clopyralid (3.02), fluroxypyr (3.70), and imazethapyr (3.90) all have relatively high GUS values(>2.8) and are thus more likely to be detected in effluent than active ingredients with smaller GUS values. This suggests that further improvements in biosystem design need to be made for optimizing the recycling of these pesticides.

Metagenomic and metatranscriptomic analyses reveal that biobed systems can enrich for antibiotic resistance and genetic mobility genes.

Antibiotic resistance gene pollution in the environment has been identified as a potential contributor to the global issue of antibiotic resistance prevalence, creating a need to identify and characterize environmental reservoirs for antibiotic resistance genes. Because many polluted environments have been shown to contain elevated levels of antibiotic resistance genes, agriculturally based pesticide bioremediation systems called 'biobeds' could serve as environmental reservoirs for antibiotic resistance genes, although this has never been extensively explored. Metagenomic and metatranscriptomic analyses of an on-farm biobed system sampled before and after a season of pesticide use demonstrated that insitu pesticide applications applied to biobeds can enrich for multidrug, sulphonamide, aminoglycoside and beta-lactam resistance genes. Additionally, this study demonstrated an enrichment for genes associated with gene mobilization, such as genes involved in horizontal gene transfer and plasmid mobility, as well as transposons and integrases.

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.

Metagenomic and metatranscriptomic analysis reveals enrichment for xenobiotic-degrading bacterial specialists and xenobiotic-degrading genes in a Canadian Prairie two-cell biobed system.

Biobeds are agriculture-based bioremediation tools used to safely contain and microbially degrade on-farm pesticide waste and rinsate, thereby reducing the negative environmental impacts associated with pesticide use. While these engineered ecosystems demonstrate efficient pesticide removal, the microbiomes in these environments remain largely understudied both taxonomically and functionally. This study used metagenomic and metatranscriptomic techniques to characterize the microbial community in a two-cell Canadian biobed system before and after a field season of pesticide application. These culture-independent approaches identified an enrichment of xenobiotic-degrading bacteria, such as Afipia, Sphingopyxis and Pseudomonas, and enrichment and transcription of xenobiotic-degrading genes, such as peroxidases, oxygenases, and hydroxylases, among others; we were able to directly link the transcription of these genes to Pseudomonas, Oligotropha, Mesorhizobium, Rhodopseudomonas, and Stenotrophomonas taxa.

Are there any risks of the disposal of pesticide effluents in soils? Biobed system meets ecotoxicology ensuring safety to soil fauna.

The biobed is a purification system, which reduces soil pollution for receiving pesticide residues from handling and washing machinery in agricultural areas. The aims of this study were (1) to assess ecotoxicity effects over time to soil fauna, posed by Lorsban® 480 BR (Chlorpyrifos) and Dithane® NT (Mancozeb) residues when disposed of in a biobed system compared with two subtropical soils, and (2) to assess ecotoxicity effects over time to soil fauna simulating an accidental spillage with Lorsban® 480 BR at the biobed. A semi-field experiment was conducted for 420 days in southern Brazil, testing continuous disposal of washing pulverization tanks in biobeds, Typic Haploperox or Typic Hapludults. In addition, different biobeds received a single dose (1 L) of Lorsban® 480 BR to simulate an accidental spillage. Chronic ecotoxicity tests were performed using Folsomia candida, Eisenia andrei, and Enchytraeus crypticus in different sampling times for both experiments. F. candida was the most sensitive species. The biobed system was able to eliminate effects from residues of both pesticides over time in all species, which did not happen in both natural soils. In accidental spillage simulation, even 420 days after contamination, F. candida did not show reproduction. The biobeds can be a feasible alternative for the disposal and treatment residues of pesticides, also for handling and washing pesticides activities. The system was efficient in promoting degradation and reducing ecotoxicity effects posed by Lorsban® 480 BR and Dithane® NT for soil fauna. It is a safe alternative to avoid soil contamination.

New analytical method for chlorpyrifos determination in biobeds constructed in Brazil: Development and validation

A quick and efficient method was optimized and validated to determine chlorpyrifos in biobeds using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). Chlorpyrifos was extracted from the matrix with 30 mL of a mixture of acetone, phosphoric acid and water 98:1:1 (v/v/v). After homogenization, centrifugation and filtration, 125 µL of the extract was evaporated and reconstituted in 5 mL of methanol acidified with 0.1% acetic acid. Validation was performed by studying analytical curve linearity (r2), estimated instrument and method limits of detection and limits of quantification (LODi, LODm, LOQi and LOQm, respectively), accuracy, precision (expressed as relative standard deviation, RSD), and matrix effect. Accuracy and precision were determined from the amount of pesticide recovered from biobed blank samples (i.e. without pesticide residue) spiked with chlorpyrifos at three different concentrations (2, 10 and 50 mg kg−1), with seven replicates at each concentration. For all three concentrations studied, the average recovery values obtained were between 96 and 115% with RSD values lower than 20%. The validated LOQ obtained was 2 mg kg−1 (from recovery studies) and the matrix effect observed was lower than ±20%, which demonstrated that there was neither considerable suppression nor enhancement of the analyte signal. The biobed system efficiently degraded chlorpyrifos in both 1) simulation of accidental spillage and 2) application of diluted pesticide solution. In the latter case, all the values obtained at the final sampling time (14 months) were below the validated LOQm.

Indigenous biobed to limit point source pollution of imidacloprid in tropical countries

Point pollution of pesticides originating from the washing of spraying machines could be controlled by biobed system and it is in use in temperate countries. The biobed system is yet to be established in tropical countries. An indigenous biobed system was prepared using local resources like rice straw, farm yard manures (FYM) and paddy field soil to suit the tropical climate. Lowermost 3 cm layer of the biobed system was filled with rice husk biochar to prevent leaching of pesticides from the system. This model system was tested with high doses of imidacloprid (178 mg/column), a commonly used pesticide against number of insect-pests in different crops, for its degradation. The bio-mix trapped a major part of the imidacloprid on the top most layer of the biobed column and only a very small part of imidacloprid recovered from the leachate. The biobed system could degrade 70.13% of applied imidacloprid within 15 days of the experiment and only 5.27% of the total pesticide recovered 90 days after incubation. Addition of biochar layer adsorbed imidacloprid from the outgoing leachate from the biobed column. Biomixture boosted microbial activity more particularly fungal population, which might be responsible for imidacloprid degradation. Microbial biomass carbon, and soil enzymes indicated faster dissipation of imidacloprid from the top layer of the biobed. This simple but efficient biobed system using local resources can fulfill the need of the small and marginal farmers of Asian countries for pesticide decontamination.

Insights into the Function and Horizontal Transfer of Isoproturon Degradation Genes (pdmAB) in a Biobed System.

Biobeds, designed to minimize pesticide point source contamination, rely mainly on biodegradation processes. We studied the interactions of a biobed microbial community with the herbicide isoproturon (IPU) to explore the role of the pdmA gene, encoding the large subunit of an N-demethylase responsible for the initial demethylation of IPU, via quantitative PCR (qPCR) and reverse transcription-PCR (RT-qPCR) and the effect of IPU on the diversity of the total bacterial community and its active fraction through amplicon sequencing of DNA and RNA, respectively. We further investigated the localization and dispersal mechanisms of pdmAB in the biobed packing material by measuring the abundance of the plasmid pSH (harboring pdmAB) of the IPU-degrading Sphingomonas sp. strain SH (previously isolated from the soil used in the biobed) compared with the abundance of the pdmA gene and metagenomic fosmid library screening. pdmA abundance and expression increased concomitantly with IPU mineralization, verifying its major role in IPU transformation in the biobed system. DNA- and RNA-based 16S rRNA gene sequencing analysis showed no effects on bacterial diversity. The pdmAB-harboring plasmid pSH showed a consistently lower abundance than pdmA, suggesting the localization of pdmAB in replicons other than pSH. Metagenomic analysis identified four pdmAB-carrying fosmids. In three of these fosmids, the pdmAB genes were organized in a well-conserved operon carried by sphingomonad plasmids with low synteny with pSH, while the fourth fosmid contained an incomplete pdmAB cassette localized in a genomic fragment of a Rhodanobacter strain. Further analysis suggested a potentially crucial role of IS6 and IS256 in the transposition and activation of the pdmAB operon.IMPORTANCE Our study provides novel insights into the interactions of IPU with the bacterial community of biobed systems, reinforces the assumption of a transposable nature of IPU-degrading genes, and verifies that on-farm biobed systems are hot spots for the evolution of pesticide catabolic traits.

Bacteria amended clay biochar composite biobed system to treat agriculture runoff

An efficient adsorbent which can resolve the existing limitations of a biobed is of concern. In the present study, a composite is prepared by mixing and pyrolyzing clay and plant parts. This is finally converted to clay biochar composite with enhanced porosity and adsorption capacity. Composite consists of clay with sawdust or clay with powdered dry fruit of Acacia concinna. Among the different composites employed, clay/Acacia concinna (7.6/0.4) with higher structural stability was used as the biomix for biobed. The clay biochar composite (20%) bioaugmented with biosurfactant producing bacterial consortium was then mixed with sandy clay loam soil in a laboratory-scale biobed system. The study showed a COD removal of 95% and cypermethrin removal of 98%. Biodegradation of cypermethrin isomers in soil and clay biochar composite was observed. The study revealed that clay biochar composite amended with biosurfactant producing bacterial consortium is an efficient biomix for the biobed system.

Atrazine dissipation in a biobed system inoculated with immobilized white-rot fungi

ABSTRACTDue to environmental concerns about accumulation of the herbicide atrazine in food products and water reservoirs, there is a need to develop safe and economical methods for its dissipation. The main aim of this study was to evaluate atrazine dissipation in a biobed system inoculated with immobilized white-rot fungi in a pelletized support (PS). All fungal isolates evaluated were efficient in colonizing the surface and inner parts of the PS, with no differences observed in the colonization over the assay. The highest atrazine dissipation (93%) was observed for Stereum hirsutum Ru-104 after 60 days of incubation. In contrast, the lowest atrazine dissipation (78%) was observed for the non-inoculated biobed over an identical period. Trametes versicolor HL01 was involved in the highest phenoloxidase activity. This activity indicates that inoculated and non-inoculated biobeds were biologically active over the incubation period. A high respiration rate in the biobed was observed when it was inoculated with S. hirsutum Ru-104. Inoculation of fungi on PS systems may be a coadjutant in the increase of fungal efficiency in enzymatic production.

Rhizosphere effect on pesticide degradation in biobeds under different hydraulic loads

Interactions between microorganisms and root exudates in a biobed system with vegetal (grass) cover could enhance pesticide degradation. Otherwise, a high water load may generate high concentrations of pesticides in lixiviates. We studied the effect of the vegetal cover on the degradation of a mixture of atrazine (ATZ), chlorpyrifos (CHL) and iprodione (IPR) (35 mg L -1 each) in a biobed system operated with two different hydraulic loads (0.6 and 1.2 L of tap water per day). The concentration of the pesticides and their main metabolites were measured in the lixiviates during 60 days, as well as in the biomixtures at the end of the study. Dehydrogenase activity in the biomixtures and organic acid exudation from the vegetal cover were also analysed. The vegetal cover diminished the lixiviation of pesticides and their metabolites mainly at the lower hydraulic load used. The degradation of the pesticides was high (>95%) and increased in biobeds with vegetal cover and low hydraulic load. Degradation metabolites of CHL and IPR were formed during pesticide degradation; however they were degraded in the biobed and were not detected in lixiviates at the end of the study. In general, an increase in organic acid exudation by vegetal cover was observed caused by chemical stress after pesticide application. The increase was similar at both hydraulic loads. Efficient colonisation of wheat straw by fungi was observed by confocal microscopy.

Evaluation of spent mushroom substrate as substitute of peat in Chinese biobeds

Biobeds are an effective system for minimising the risk of point-source contamination by pesticides. The original biobed system comprises a clay layer at the bottom, a biomixture layer and a grass layer on top. The composition of the biomixture, which in the original Swedish model consisted of soil: peat: wheat straw (1:1:2 by volume), determines the efficiency of the system. Since the use of peat is not environmentally or economically feasible in China, this study tested the potential of replacing it with a locally available material, spent mushroom substrate (SMS). Three biomixtures containing different SMS (Pleurotus eryngii, Flammulina velutipes and Lentinus edodes) were compared with a biomixture containing peat, as in the original Swedish design, and a control containing soil alone. The fungicide chlorothalonil and the insecticide imidacloprid were used as model pesticides in the tests. Microbial activity (measured as respiration and phenoloxidase and hydrolytic activity) and pesticide dissipation were studied. Microbial activity was higher in the three biomixtures containing SMS than in the original-type biomixture. Among the SMS biomixtures, that containing SMS from L. edodes was the most biologically active. However, pesticide dissipation was comparable in all four biomixtures and significant differences were only found between biomixtures and the soil-alone control. Based on the physicochemical characteristics, biological activity and preliminary results on pesticide degradation, SMS are suitable and can therefore be used as a substitutes for peat.

Using volatile organic compounds to enhance atrazine biodegradation in a biobed system

The effect of the terpenes α-pinene, eucalyptol, and limonene, individually and as mixtures, on atrazine (ATZ) biodegradation and on biological activity in a biobed biomixture was evaluated. Additionally, terpenes emitted from the biomixture were captured using solid-phase microextraction. Terpenes added individually at relatively low concentrations (50μgkg(-1)) significantly enhanced ATZ degradation and biological activity during the first incubation days. No significant effect on ATZ degradation was found from adding the terpene mixture, and, interestingly, an inhibitory effect on phenoloxidase activity was found during the first 20days of incubation when mixed terpenes were present at 100μgkg(-1). Capturing terpenes demonstrated that during the first hour of incubation a significant fraction of the terpenes was volatilized. These results are the first to demonstrate the feasibility of using terpenes to enhance the degradation of a pesticide. However, successive applications of terpenes or the addition of materials that slowly release terpenes could sustain the ATZ degradation enhancement.

Atrazine degradation in the biomixture of a biobed system biostimulated with terpenes

Atrazine degradation in the biomixture of a biobed system biostimulated with terpenes

Degradation and Adsorption of Pesticides in Compost-Based Biomixtures as Potential Substrates for Biobeds in Southern Europe

Biobeds have been used in northern Europe for minimizing point source contamination of water resources by pesticides. However, little is known regarding their use in southern Europe where edaphoclimatic conditions and agriculture practices significantly differ. A first step toward their adaptation in southern Europe is the use of low-cost and easily available substrates as biomixture components. This study investigated the possibility of replacing peat with agricultural composts in the biomixture. Five composts from local substrates including olive leaves, cotton crop residues, cotton seeds, spent mushroom substrate, and commercial sea wrack were mixed with topsoil and straw (1:1:2). Degradation of a mixture of pesticides (dimethoate, indoxacarb, buprofezin, terbuthylazine, metribuzin, metalaxyl-M, iprodione, azoxystrobin) at two dose rates was tested in the compost biomixtures (BX), in corresponding peat biomixtures (OBX), and in soil. Adsorption-desorption of selected pesticides were also studied. Pesticide residues were determined by gas chromatography with nitrogen-phosphorus detector, except indoxacarb, which was determined with a microelectron capture detector. Overall, BX degraded the studied pesticides at rates markedly higher than those observed in soil and OBX, in which the slowest degradation rates were evident. Overall, the olive leaf compost biomixture showed the highest degradation capacity. Adsorption studies showed that OBX and BX had higher adsorption affinity compared to soil. Desorption experiments revealed that pesticide adsorption in biomixtures was not entirely reversible. The results suggest that substitution of peat with local composts will lead to optimization of the biobed system for use in Mediterranean countries.

BIOSTIMULATION OF AGRICULTURAL BIOBEDS WITH NPK FERTILIZER ON CHLORPYRIFOS DEGRADATION TO AVOID SOIL AND WATER CONTAMINATION

Degradation of the insecticide chlorpyrifos (160 a.i mg kg -1 ) using a biomix of a biobed system biostimulated with inorganic fertilizer (NPK) was investigated. Three concentrations of the fertilizer (0.1%, 0.5% and 1.0% ww -1 ) were evaluated on chlorpyrifos degradation, TCP (3, 5, 6-trichloro-2-pyrinidol) accumulation and biological activity of the biomix. The chlorpyrifos was dissipated efficiently (>75%) after 40 days of incubation and no additional dissipation was obtained with increasing concentration of NPK after 20 days of incubation. TCP accumulation occurred in all evaluated NPK concentrations and its concentration increased with the increment of NPK addition raising the probability of leaching of this compound. Biological activity (FDA and ligninolytic enzyme activity) in the biomix increased by the NPK presence in all evaluated concentrations. The DGGE analyses showed that combined treatments with lower amounts of NPK (0% and 0.1%) and chlorpyrifos showed no significant modifications in the microbial community in the biomix. However, combined overdoses of NPK (0.5 and 1.0%) and chlorpyrifos caused significant modifications in the bacterial communities that could be associated with TCP degradation reduction in the biomix. In conclusion, the obtained results demonstrated that the biomix prepared with Andisol and biostimulated with NPK nutrient can be recommended in biobeds as a viable alternative of chlorpyrifos dissipation avoiding soil and water contamination probability.

Biobeds for Environmental Protection from Pesticide Use—A Review

Biobeds originated in Sweden in response to the need for simple and effective methods to minimize environmental contamination from pesticide use, especially when filling spraying equipment, a typical point source of contamination. The biobed system has attracted attention in several countries, where work is being conducted to adapt it to local conditions and applications. As a consequence, the biobed system has been more or less modified and sometimes renamed, for example, as biomassbed in Italy, biofilter in Belgium, and Phytobac and biobac in France. The effectiveness and simplicity of the biobed also make it suitable for use in developing countries, and different adaptations of the biobed concept now exist in, for instance, Peru, Guatemala, and Ecuador. When the modification of the biobed includes an intention to use it for retention and degradation of pesticides in sprayer washings, the construction has to be adapted to, for example, lined biobeds to ensure that no pesticide leaching will occur. Replacement of some of the original materials in the Swedish biomixture (straw, peat, and soil) can also change the performance of the system, for instance, the amount, activity, and composition of the microbial community that develops. This review presents the state of the art of biobeds and similar systems in Sweden and worldwide and identifies future research needs. Factors affecting the efficiency of biobeds in terms of degradation and retention of pesticides are discussed, with particular emphasis on the microbial processes involved.

Degradation and leaching potential of pesticides in biobed systems.

Biobeds provide a potential solution to pesticide contamination of surface waters arising from the farmyard. Previous work has shown that biobeds can effectively treat spills and splashes of pesticide. This study investigated the potential for biobeds to treat much larger volumes and amounts of pesticide waste not only arising from spills but also from washing processes. Two systems were assessed using a range of pesticides at the semi-field scale, ie a lined biobed system and an unlined system. Studies using the lined biobeds demonstrated that water management was crucial, with biobeds needing to be covered to exclude rain-water. Once covered, the top of the biobed became hydrophobic, restricting moisture loss and resulting in saturated conditions at depth. The drying out of the top layer coincided with a measured decrease in microbial biomass in the treated biobeds. Applied pesticides were effectively retained within the 0-5 cm layer. Whilst all pesticides tested degraded, low moisture content and microbial activity meant degradation rates were low. Studies using unlined biobeds showed that only the most mobile pesticides leached, and for these > 99% was removed by the system, with a significant proportion degraded within 9 months. Peak concentrations of the two most mobile pesticides did however exceeded the limits that are likely to be required by regulatory bodies. However, it is thought that these limits could be reached by optimisation of the system.