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1.
Front Plant Sci ; 13: 1009956, 2022.
Article in English | MEDLINE | ID: mdl-36426148

ABSTRACT

Soil biosolarization (SBS) is an alternative technique for soil pest control to standard techniques such as soil fumigation and soil solarization (SS). By using both solar heating and fermentation of organic amendments, faster and more effective control of soilborne pathogens can be achieved. A circular economy may be created by using the residues of a given crop as organic amendments to biosolarize fields that produce that crop, which is termed circular soil biosolarization (CSBS). In this study, CSBS was employed by biosolarizing soil with amended tomato pomace (TP) residues and examining its impact on tomato cropping under conditions of abiotic stresses, specifically high salinity and nitrogen deficiency. The results showed that in the absence of abiotic stress, CSBS can benefit plant physiological performance, growth and yield relative to SS. Moreover, CSBS significantly mitigated the impacts of abiotic stress conditions. The results also showed that CSBS impacted the soil microbiome and plant metabolome. Mycoplana and Kaistobacter genera were found to be positively correlated with benefits to tomato plants health under abiotic stress conditions. Conversely, the relative abundance of the orders RB41, MND1, and the family Ellin6075 and were negatively correlated with tomato plants health. Moreover, several metabolites were significantly affected in plants grown in SS- and CSBS-treated soils under abiotic stress conditions. The metabolite xylonic acid isomer was found to be significantly negatively correlated with tomato plants health performance across all treatments. These findings improve understanding of the interactions between CSBS, soil ecology, and crop physiology under abiotic stress conditions.

2.
Bioprocess Biosyst Eng ; 45(2): 333-344, 2022 Feb.
Article in English | MEDLINE | ID: mdl-34862916

ABSTRACT

Global demand for poultry and associated feed are projected to double over the next 30 years. Insect meal is a sustainable alternative to traditional feeds when produced on low-value high-volume agricultural byproducts. Black soldier fly (BSF) larvae (Hermetia illucens L.) are high in protein and contain methionine, an essential amino acid that is critical to poultry health. BSF larvae can be grown on many organic residues, however, larvae growth and quality vary based on feedstock and cultivation processes. Experiments were completed to monitor temporal changes in BSF larvae growth and composition using almond hulls as a growth substrate under batch and semi-batch processes and with varying substrate carbon to nitrogen ratio (C/N). A logistic kinetic growth model was developed to predict larval biomass and methionine accumulations during batch production. Estimated ranges of model parameters for larvae maximum specific growth rate and carrying capacity were 0.017-0.021 h-1 and 9.7-10.7 g larvae kg-1 hulls dry weight, respectively. Methionine content in larvae increased from 11.1 to 17.1 g kg-1 dry weight over a 30-day batch incubation period. Larvae-specific growth and yield increased by 168% and 268%, respectively, when cultivated in a semi-batch compared to a batch process. Increasing C/N ratio from 26 to 40 increased density of methionine content in larvae per unit feedstock by 25%. The findings demonstrate a logistic model can predict larvae biomass accumulation, harvest time can achieve specific methionine contents, and a semi-batch process is more favorable for larvae biomass accumulation compared to a batch process.


Subject(s)
Animal Feed , Diptera , Animal Feed/analysis , Animals , Biomass , Diptera/metabolism , Larva , Methionine/metabolism
3.
J Ind Microbiol Biotechnol ; 47(12): 1031-1044, 2020 Dec.
Article in English | MEDLINE | ID: mdl-33128103

ABSTRACT

The properties of digestates generated through anaerobic digestion are influenced by interactions between the digester microbial communities, feedstock properties and digester operating conditions. This study investigated the effect of varying initial feedstock carbon to nitrogen (C/N) ratios on digestate microbiota and predicted abundance of genes encoding lignocellulolytic activity. The C/N ratio had a significant impact on the digestate microbiome. Feedstocks with intermediate C/N ratio (20-27) (where higher biomethane potential was observed) showed higher relative abundance of archaea compared to feedstocks with C/N ratios at 17 and 34. Within microbial networks, four microbial clusters and eight connector microorganisms changed significantly with the C/N ratio (P < 0.05). Feedstocks with C/N < 23 were richer in organisms from the family Thermotogaceae and genus Caldicoprobacter and enhanced potential for degradation of maltose, galactomannans, melobiose and lactose. This study provides new insights into how anaerobic digestion conditions relate to the structure and functional potential of digester microbial communities, which may be relevant to both digester performance and subsequent utilization of digestates for composting or amending soil.


Subject(s)
Anaerobiosis , Soil Microbiology , Archaea , Bacteria , Bioreactors , Carbon , Composting , Food , Microbiota , Nitrogen/chemistry , Refuse Disposal , Soil/chemistry
4.
PLoS One ; 15(5): e0232662, 2020.
Article in English | MEDLINE | ID: mdl-32369503

ABSTRACT

Regulatory pressure along with environmental and human health concerns drive the development of soil fumigation alternatives such as soil biosolarization (SBS). SBS involves tarping soil that is at field capacity with a transparent film following amendment with certain organic materials. Heating via the greenhouse effect results in an increase of the soil temperature. The organic amendments can promote microbial activity that can enhance pest inactivation by depleting oxygen, producing biopesticidal fermentation products, and competing with pests. The properties of the organic amendments can heavily influence the type and magnitude of these effects. This study evaluated the viability of chitin as a novel SBS soil amendment to influence soil fungal and bacterial microbial communities, including control of the plant pathogen Fusarium oxysporum f.sp. lactucae (FOL). Changes to FOL and the broader soil microbiota were monitored in response to biosolarization using 0.1% (by dry weight) amendment with chitin (Rootguard). FOL suppression was only observed in chitin amended soils that were incubated at room temperature, not under solarized conditions. Conversely, it decreased solarization efficacy in the upper (0-10 cm) soil layer. The presence of chitin also showed increase in FOL under anaerobic and fluctuating temperature regime conditions. Biosolarization with chitin amendment did exhibit an impact on the overall soil microbial community. The fungal genus Mortierella and the bacterial family Chitinophagaceae were consistently enriched in biosolarized soils with chitin amendment. This study showed low potential FOL suppression due chitin amendment at the studied levels. However, chitin amendment showed a higher impact on the fungal community than the bacterial community. The impact of these microbial changes on crop protection and yields need to be studied in the long-term.


Subject(s)
Chitin/chemistry , Composting/methods , Fusarium/growth & development , Soil Microbiology , Soil/chemistry , Crop Production , Microbiota , Pest Control, Biological
5.
J Sci Food Agric ; 100(12): 4618-4626, 2020 Sep.
Article in English | MEDLINE | ID: mdl-32419145

ABSTRACT

BACKGROUND: Insect biomass is a sustainable alternative to traditional animal feeds, particularly when insects are produced on low-value high-volume agricultural by-products. Seven samples of almond by-product (hulls and shells) were obtained from processors in California and investigated for larvae production. Experiments were completed with and without larvae and spent substrate samples were assessed for their potential as soil amendments based on standard compost quality indicators. RESULTS: On average, specific larvae growth and average larval harvest weight were 158% and 109% higher, respectively, when larvae were reared on Monterey and pollinator hulls compared to nonpareil hulls and mixed shells. Larvae methionine and cystine contents were highest when larvae were reared on Monterey hulls and mixed shells, respectively. Available phytonutrients in spent substrate were affected by feedstock sample and larvae rearing. Spent nonpareil substrate without larvae had the highest NH4 -N levels and spent pollinator substrate incubated without larvae had the highest PO4 -P levels. Spent mixed shell substrate had the lowest availability of phytonutrients. CONCLUSION: The findings demonstrate that by-product composition has a significant impact on larvae growth and the properties of the spent substrate, and that spent substrate from larvae rearing requires further stabilization before application as a soil amendment. © 2020 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Subject(s)
Animal Feed/analysis , Diptera/metabolism , Prunus dulcis/chemistry , Prunus dulcis/metabolism , Waste Products/analysis , Animals , Diptera/growth & development , Larva/growth & development , Larva/metabolism , Seeds/chemistry , Seeds/metabolism , Soil/chemistry
6.
Biotechnol Bioeng ; 117(1): 62-72, 2020 01.
Article in English | MEDLINE | ID: mdl-31531975

ABSTRACT

Wastewater treatment is an energy-intensive process and a net emitter of greenhouse gas emissions. A large fraction of these emissions is due to intensive aeration of aerobic bacteria to facilitate break-down of organic compounds. Algae can generate dissolved oxygen at levels in excess of saturation, and therefore hold the potential to partially displace or complement mechanical aeration in wastewater treatment processes. The objective of this study was to develop an internally consistent experimental and modeling approach to test the hypothesis that algal photosynthetic aeration can speed the removal of organic constituents by bacteria. This framework was developed using a simplified wastewater treatment process consisting of a model bacteria (Escherichia coli), a model algae (Auxenochlorella protothecoides), and a single carbon source that was consumable by bacteria only. This system was then tested both with and without the presence of algae. A MATLAB model that considered mass transfer and biological kinetics was used to estimate the production and consumption of O2 and CO2 by algae and bacteria. The results indicated that the presence of algae led to 18-66% faster removal of COD by bacteria, and that roughly one-third of biochemical oxygen demand was offset by algal photosynthetic aeration.


Subject(s)
Bacteria/metabolism , Chlorophyta/metabolism , Oxygen/metabolism , Wastewater/chemistry , Water Purification/methods , Biological Oxygen Demand Analysis , Bioreactors/microbiology , Carbon/metabolism , Models, Biological , Photosynthesis/physiology , Wastewater/microbiology
7.
Waste Manag ; 101: 74-82, 2020 Jan 01.
Article in English | MEDLINE | ID: mdl-31604160

ABSTRACT

Biosolarization utilizes organic amendments to produce biopesticide compounds in soil that can work in tandem with other stresses to inactivate agricultural pests. The prospect of using by-products from industrial almond processing as amendments for biosolarization was assessed. Soil mesocosms were used to simulate biosolarization using various almond by-products, application rates, and incubation times. Several potentially biopesticidal organic acids were identified and quantified in the soil, and the toxicity of soil extracts was evaluated for the root lesion nematode (Pratylenchus vulnus). It was determined that both almond hulls and a mixture of hulls and shells harbored several acids, the concentration of which was enhanced 1-7 fold via fermentation by native soil microbes. Organic acid concentration in the soil showed a significant linear relationship with the quantity of waste biomass amended. Extracts from soils containing at least 2.5% incorporated biomass by dry weight showed a 84-100% mortality of nematodes, which corresponded to acid concentrations 0.75 mg/g (2.0 g/L) or greater. This study showed that almond processing by-products - hulls and a hull and shell mixture - were suitable amendments for control of P. vulnus and potentially other soil agricultural pests in the context of biosolarization.


Subject(s)
Biological Control Agents , Prunus dulcis , Agriculture , Biomass , Soil
8.
Appl Microbiol Biotechnol ; 104(2): 893-905, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31828407

ABSTRACT

The use of microalgae for wastewater treatment has been proposed as a cost-effective method to produce biofuels while remediating waste streams. This study examined the microalgae biomass production rate, wastewater treatment efficiency, and prokaryotic organism microbiome associated with microalgae Chlorella sorokiniana cultivated on anaerobic digestate effluent. Final microalgae biomass concentrations from nine photobioreactors were highly variable and had values that ranged between 0.14 g/L and 0.90 g/L. Nutrient removal efficiencies for TN (total nitrogen), N-NH4 (ammonium nitrogen), and COD (chemical oxygen demand) ranged from 34% to 67%, 65% to 97%, and-60% to 14%, respectively. Analysis of individual OTUs (operational taxonomic units) from the microbial community revealed that microalgae biomass concentrations were significantly correlated with the relative abundance of OTUs in the genus Pusillimonas. Predictive metagenomic analyses identified additional correlations associated with biomass production and nutrient removal. These results suggest that the microbial community present during microalgae cultivation on wastewater can impact the performance of the system for biomass production and wastewater treatment.


Subject(s)
Bacteria/growth & development , Bacteria/metabolism , Microalgae/growth & development , Microalgae/metabolism , Microbiota , Wastewater/microbiology , Water Purification/methods , Ammonium Compounds/analysis , Bacteria/classification , Biological Oxygen Demand Analysis , Bioreactors/microbiology , Metagenomics , Microalgae/classification , Nitrogen/analysis , Wastewater/chemistry , Water Pollutants/metabolism
9.
NPJ Sci Food ; 3: 15, 2019.
Article in English | MEDLINE | ID: mdl-31508493

ABSTRACT

Increases in global human population are leading to increasing demands for food production and waste management. Insect biomass is a sustainable alternative to traditional animal feeds when insects are produced on lignocellulosic by-products. Resources high in lignocellulose have high carbon to nitrogen ratios and require nitrogen supplementation to accelerate bioconversion. Here we report on studies that examine the influence of nitrogen supplementation of almond hull-based feedstocks on black soldier fly larvae (Hermetia illucens L.) cultivation and composition. Decreasing carbon to nitrogen ratio from 49 to 16 increased larvae harvest dry weight, specific larvae growth, and yield by 36%, 31%, and 51%, respectively. However, the decrease in carbon to nitrogen ratio decreased larvae methionine and cysteine contents by 11% and 13%, respectively. The findings demonstrate that carbon to nitrogen ratio can be managed to enhance bioconversion of lignocellulose to larvae, but that this management approach can reduce larvae amino acid content.

10.
J Sci Food Agric ; 98(15): 5893-5900, 2018 Dec.
Article in English | MEDLINE | ID: mdl-29999178

ABSTRACT

BACKGROUND: The increasing production of almonds worldwide has resulted in the significant generation of byproduct streams that require end uses. One potential use for byproducts is for cultivation of additional food sources including insects. Studies were performed to determine if black soldier fly larvae (Hermetia illucens L.) could be cultivated on almond byproducts (hulls and shells) and to examine the effect of aeration and moisture on larvae growth and hull composition. RESULTS: Increasing aeration from 0.04 to 0.36 mL min-1 g dry weight-1 tripled the harvest weight of larvae and increased larvae yield by a factor of five. Larvae calcium content increased by 18% with an increase in aeration from 0.04 to 0.95 mL min-1 g dry weight-1 . Moisture content also affected harvest dry weight and yield; increasing moisture content from 480 g kg-1 (wet basis) to 680 g kg-1 increased harvest weight by 56% and yield by a factor of 2. Variables did not affect larvae methionine and cysteine content. Low moisture content and aeration rate yielded an environment that supported microbial consumption of hulls over larvae consumption and growth. CONCLUSIONS: The results demonstrate that almond hulls are a suitable feedstock for larvae production under controlled management of moisture content and aeration. © 2018 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Subject(s)
Larva/growth & development , Prunus dulcis/parasitology , Simuliidae/growth & development , Waste Products/analysis , Animal Feed/analysis , Animals , Larva/metabolism , Nuts/chemistry , Nuts/metabolism , Nuts/parasitology , Prunus dulcis/chemistry , Prunus dulcis/metabolism , Simuliidae/metabolism
11.
J Agric Food Chem ; 66(22): 5451-5461, 2018 Jun 06.
Article in English | MEDLINE | ID: mdl-29763301

ABSTRACT

Conventional solarization and biosolarization with mature compost and tomato processing residue amendments were compared with respect to generation of pesticidal conditions and tomato ( Solanum lycopersicum L.) plant growth in treated soils. Soil oxygen depletion was examined as a response that has previously not been measured across multiple depths during biosolarization. For biosolarized soil, volatile fatty acids were found to accumulate concurrent with oxygen depletion, and the magnitude of these changes varied by soil depth. Two consecutive years of experimentation showed varying dissipation of volatile fatty acids from biosolarized soils post-treatment. When residual volatile fatty acids were detected in the biosolarized soil, fruit yield did not significantly differ from plants grown in solarized soil. However, when there was no residual volatile fatty acids in the soil at the time of planting, plants grown in biosolarized soil showed a significantly greater vegetation amount, fruit quantity, and fruit ripening than those of plants grown in solarized soil.


Subject(s)
Industrial Waste/analysis , Pest Control/methods , Soil/chemistry , Solanum lycopersicum/growth & development , Waste Products/analysis , Weed Control/methods , Composting , Fruit/chemistry , Fruit/growth & development , Fruit/metabolism , Solanum lycopersicum/chemistry , Solanum lycopersicum/parasitology , Plant Diseases/parasitology , Plant Diseases/prevention & control , Plant Weeds/growth & development , Plant Weeds/radiation effects , Soil/parasitology , Solar Energy
12.
Waste Manag ; 77: 555-564, 2018 Jul.
Article in English | MEDLINE | ID: mdl-29773481

ABSTRACT

The management of organic wastes is an environmental and social priority. Aerobic digestion (AED) or composting and anaerobic digestion (AD) are two organic waste management practices that produce a value-added final product. Few side-by-side comparisons of both technologies and their digestate products have been performed. The objective of this study was to compare the impact of initial feedstock properties (moisture content and/or C/N ratio) on stabilization rate by AED and AD and soil amendment characteristics of the final products. Green and food wastes were considered as they are two of the main contributors to municipal organic waste. Stabilization rate was assessed by measurement of CH4 and CO2 evolution for AD and AED, respectively. For AD, CH4 yield showed a second-order relationship with the C/N content (P < 0.05); the optimal C/N ratio indicated by the relationship was 25.5. For AED, cumulative CO2 evolution values were significantly affected by the C/N ratio and moisture content of the initial feedstock (P < 0.05). A response surface model showed optimal AED stabilization for a C/N of 25.6 and moisture of 64.9% (wet basis). AD final products presented lower soluble chemical oxygen demand (COD) but lower humification degree and aromaticity than the products from AED. This lower stability may lead to further degradation when amended to soil. The results suggest that composting feedstocks with higher C/N produces an end-product with higher suitability for soil amendment. The instability of end products from AD could be leveraged in pest control techniques that rely on organic matter degradation to produce compounds with pesticidal properties.


Subject(s)
Food , Waste Management , Anaerobiosis , Soil , Soil Microbiology
13.
J Immunol Res ; 2018: 3710961, 2018.
Article in English | MEDLINE | ID: mdl-29577048

ABSTRACT

Transgene introgression is a major concern associated with transgenic plant-based vaccines. Agroinfiltration can be used to selectively transform nonreproductive organs and avoid introgression. Here, we introduce a new vaccine modality in which Staphylococcal enterotoxin B (SEB) genes are agroinfiltrated into radishes (Raphanw sativus L.), resulting in transient expression and accumulation of SEB in planta. This approach can simultaneously express multiple antigens in a single leaf. Furthermore, the potential of high-throughput vaccine production was demonstrated by simultaneously agroinfiltrating multiple radish leaves using a multichannel pipette. The expression of SEB was detectable in two leaf cell types (epidermal and guard cells) in agroinfiltrated leaves. ICR mice intranasally immunized with homogenized leaves agroinfiltrated with SEB elicited detectable antibody to SEB and displayed protection against SEB-induced interferon-gamma (IFN-γ) production. The concept of encapsulating antigens in leaves rather than purifying them for immunization may facilitate rapid vaccine production during an epidemic disease.


Subject(s)
Enterotoxins/genetics , Plant Epidermis/genetics , Plant Extracts/immunology , Plant Leaves/genetics , Raphanus , Staphylococcus aureus/genetics , Vaccines/immunology , Animals , Antibodies, Bacterial/blood , Cells, Cultured , Enterotoxins/immunology , Immunity, Humoral , Mice , Mice, Inbred ICR , Plants, Genetically Modified/genetics , Protein Engineering , Staphylococcus aureus/immunology
14.
Pest Manag Sci ; 74(8): 1892-1902, 2018 Aug.
Article in English | MEDLINE | ID: mdl-29446871

ABSTRACT

BACKGROUND: Soil biosolarization is a promising alternative to conventional fumigation. Volatile fatty acids (VFAs) produced in the soil through fermentation of amended organic matter can affect pest inactivation during biosolarization. The objective was to determine how soil amended with organic wastes that were partially stabilized through either composting or anaerobic digestion affected the inactivation of Brassica nigra (BN; a weed) and Fusarium oxysporum f. sp. lactucae (FOL; a phytopathogenic fungus). RESULTS: The mortality of BN seeds in the biosolarized soil was 12% higher than in the solarized soil, although this difference was not significant. However, a significant correlation between BN mortality and VFA accumulation was observed. The number of FOL colony-forming units (CFU) in solarized samples at 5 cm was 34 CFU g-1 of soil, whereas in the biosolarized samples levels were below the limit of quantification. At 15 cm, these levels were 100 CFU g-1 for solarized samples and < 50 CFU g-1 of soil for the biosolarized samples. Amendment addition positively affected the organic matter and potassium content after the solarization process. CONCLUSION: The organic waste stabilization method can impact downstream biosolarization performance and final pest inactivation levels. This study suggests that organic waste management practices can be leveraged to improve pest control and soil quality. © 2018 Society of Chemical Industry.


Subject(s)
Composting/methods , Fusarium/radiation effects , Mustard Plant/radiation effects , Pest Control/methods , Soil/chemistry , Sunlight , Anaerobiosis , Bioreactors , Fusarium/physiology , Mustard Plant/physiology , Plant Weeds/physiology , Plant Weeds/radiation effects , Waste Management/methods , Weed Control/methods
15.
J Agric Food Chem ; 65(17): 3434-3442, 2017 May 03.
Article in English | MEDLINE | ID: mdl-28409935

ABSTRACT

Anaerobic digestion is an organic waste bioconversion process that produces biofuel and digestates. Digestates have potential to be applied as soil amendment to improve properties for crop production including phytonutrient content and pest load. Our objective was to assess the impact of solid anaerobic digestates on weed seed inactivation and soil quality upon soil biosolarization (a pest control technique that combines solar heating and amendment-induced microbial activity). Two solid digestates from thermophilic (TD) and mesophilic (MD) digesters were tested. The solarized TD-amended samples presented significantly higher mortality of Brassica nigra (71%, P = 0.032) than its equivalent incubated at room temperature. However, biosolarization with digestate amendment led to decreased weed seed mortality in certain treatments. The plant-available water, total C, and extractable P and K were significantly increased (P < 0.05) in the incubated amended soils. The results confirm the potential of digestates as beneficial soil amendments. Further studies are needed to elucidate the impacts of digestate stability on biosolarization efficacy and soil properties.


Subject(s)
Agriculture/methods , Seeds/chemistry , Soil/chemistry , Agriculture/instrumentation , Anaerobiosis , Biofuels/analysis , Mustard Plant/chemistry , Mustard Plant/physiology , Mustard Plant/radiation effects , Plant Weeds/chemistry , Plant Weeds/physiology , Plant Weeds/radiation effects , Seeds/radiation effects , Sunlight
16.
Pest Manag Sci ; 73(5): 862-873, 2017 May.
Article in English | MEDLINE | ID: mdl-27391139

ABSTRACT

BACKGROUND: Biosolarization is a fumigation alternative that combines passive solar heating with amendment-driven soil microbial activity to temporarily create antagonistic soil conditions, such as elevated temperature and acidity, that can inactivate weed seeds and other pest propagules. The aim of this study was to use a mesocosm-based field trial to assess soil heating, pH, volatile fatty acid accumulation and weed seed inactivation during biosolarization. RESULTS: Biosolarization for 8 days using 2% mature green waste compost and 2 or 5% tomato processing residues in the soil resulted in accumulation of volatile fatty acids in the soil, particularly acetic acid, and >95% inactivation of Brassica nigra and Solanum nigrum seeds. Inactivation kinetics data showed that near complete weed seed inactivation in soil was achieved within the first 5 days of biosolarization. This was significantly greater than the inactivation achieved in control soils that were solar heated without amendment or were amended but not solar heated. CONCLUSION: The composition and concentration of organic matter amendments in soil significantly affected volatile fatty acid accumulation at various soil depths during biosolarization. Combining solar heating with organic matter amendment resulted in accelerated weed seed inactivation compared with either approach alone. © 2016 Society of Chemical Industry.


Subject(s)
Plant Weeds/radiation effects , Seeds/radiation effects , Soil , Solanum lycopersicum/chemistry , Sunlight , Waste Management/methods , Fatty Acids/analysis , Fatty Acids/chemistry , Hot Temperature , Hydrogen-Ion Concentration , Plant Weeds/physiology , Seeds/physiology , Soil/chemistry , Temperature , Volatilization
17.
mSystems ; 1(6)2016.
Article in English | MEDLINE | ID: mdl-27981239

ABSTRACT

Ionic liquid (IL) pretreatment is a promising approach for the conversion of lignocellulose to biofuels. The toxicity of residual IL, however, negatively impacts the performance of industrial enzymes and microorganisms in hydrolysis and fermentation. In this study, a thermophilic microbial community was cultured on switchgrass amended with various levels of the ionic liquid 1-ethyl-3-methylimidazolium acetate. Changes in the microbial community composition and transcription of genes relevant to IL tolerance and lignocellulose hydrolysis were quantified. Increasing the level of IL to 0.1% (wt) led to increased levels of relative abundance and transcription in organisms of the phylum Firmicutes. Interestingly, IL concentrations of up to 1% (wt) also resulted in greater xylanase transcription and enzyme activity as well as increased transcription of endoglucanase, beta-glucosidase, and IL tolerance genes compared to communities without IL. IL levels above 1% (wt) resulted in decreased enzyme activity and transcription of genes involved in lignocellulose hydrolysis. The results indicate that moderate levels of IL select for thermophilic microorganisms that not only tolerate IL but also effectively hydrolyze lignocellulose from switchgrass. Discovery of IL-tolerant organisms and enzymes is critical for the development of biological processes that convert IL-pretreated biomass to biofuels and chemicals. Employing metatranscriptomic analysis of enrichment cultures can facilitate the discovery of microorganisms and enzymes that may be active in the presence of toxic compounds such as ionic liquids. IMPORTANCE Pretreatment using ionic liquids (IL) is a promising approach for the conversion of lignocellulose to biofuels. Because IL can be inhibitory to enzymes and microorganisms involved in downstream hydrolysis and fermentation steps, discovery of IL-tolerant organisms and enzymes is critical for advancing this technology. Employing metatranscriptomics in the analysis of IL-enriched cultures facilitated tracking of dynamic changes in a complex microbial community at the level of gene transcription and doing so with genome resolution. Specific organisms were discovered that could simultaneously tolerate a moderate IL concentration and transcribe a diverse array of cellulolytic enzymes. Gene sequences of cellulolytic enzymes and efflux pumps from those same organisms were also identified, providing important resources for future research on engineering IL-tolerant organisms and enzymes.

18.
Appl Microbiol Biotechnol ; 100(24): 10237-10249, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27838839

ABSTRACT

Chemical and physical pretreatment of biomass is a critical step in the conversion of lignocellulose to biofuels and bioproducts. Ionic liquid (IL) pretreatment has attracted significant attention due to the unique ability of certain ILs to solubilize some or all components of the plant cell wall. However, these ILs inhibit not only the enzyme activities but also the growth and productivity of microorganisms used in downstream hydrolysis and fermentation processes. While pretreated biomass can be washed to remove residual IL and reduce inhibition, extensive washing is costly and not feasible in large-scale processes. IL-tolerant microorganisms and microbial communities have been discovered from environmental samples and studies begun to elucidate mechanisms of IL tolerance. The discovery of IL tolerance in environmental microbial communities and individual microbes has lead to the proposal of molecular mechanisms of resistance. In this article, we review recent progress on discovering IL-tolerant microorganisms, identifying metabolic pathways and mechanisms of tolerance, and engineering microorganisms for IL tolerance. Research in these areas will yield new approaches to overcome inhibition in lignocellulosic biomass bioconversion processes and increase opportunities for the use of ILs in biomass pretreatment.


Subject(s)
Biological Products/metabolism , Drug Resistance, Microbial , Ionic Liquids/toxicity , Lignin/metabolism , Microbial Consortia , Solvents/toxicity , Biofuels , Biotransformation
19.
Appl Microbiol Biotechnol ; 100(12): 5639-52, 2016 Jun.
Article in English | MEDLINE | ID: mdl-27102129

ABSTRACT

The aims of this study were to identify thermophilic microbial communities that degrade green waste in the presence of the ionic liquids (IL) tetrabutylphosphonium chloride and tributylethylphosphonium diethylphosphate and examine preservation methods for IL-tolerant communities. High-solids incubations with stepwise increases in IL concentration were conducted to enrich for thermophilic IL-tolerant communities that decomposed green waste. 16S rRNA sequencing of enriched communities revealed microorganisms capable of tolerating high levels of IL. Cryogenic preservation of enriched communities reduced the IL tolerance of the community and decreased the relative abundance of IL-tolerant organisms. The use of cryoprotectants did not have an effect on microbial activity on green waste of the stored community. A successful approach was developed to enrich communities that decompose green waste in thermophilic high-solids environments in the presence of IL. Alternative community storage and revival methods are necessary for maintenance and recovery of IL-tolerant communities. The enriched communities provide a targeted source of enzymes for the bioconversion of IL-pretreated green waste for conversion to biofuels.


Subject(s)
Biofuels , Ionic Liquids/pharmacology , Microbial Consortia/drug effects , Organophosphorus Compounds/pharmacology , Biomass , Cryoprotective Agents , Drug Tolerance , Imidazoles , Microbial Consortia/physiology , RNA, Ribosomal, 16S , Waste Disposal, Fluid/methods
20.
Waste Manag ; 48: 156-164, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26525530

ABSTRACT

Pomaces from tomato paste and wine production are the most abundant fruit processing residues in California. These residues were examined as soil amendments for solarization to promote conditions conducive to soil disinfestation (biosolarization). Simulated biosolarization studies were performed in both aerobic and anaerobic soil environments and soil temperature elevation, pH, and evolution of CO2, H2 and CH4 gases were measured as metrics of soil microbial activity. Tomato pomace amendment induced conditions associated with soil pest inactivation, including elevation of soil temperature by up to 2°C for a duration of 4days under aerobic conditions and a reduction of soil pH from 6.5 to 4.68 under anaerobic conditions. White wine grape pomace amendment showed similar trends but to a lesser extent. Red wine grape pomace was generally less suitable for biosolarization due to significantly lower soil temperature elevations, reduced acidification relative to the other pomaces and induction of methanogenesis in the soil.


Subject(s)
Food-Processing Industry , Soil , Solanum lycopersicum , Waste Products , California , Ecotoxicology/methods , Germination , Hydrogen-Ion Concentration , Lactuca/growth & development , Oxygen/analysis , Soil/chemistry , Temperature , Vitis , Wine
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