Detoxification of corn stover prehydrolysate by different biochars and its effect on lactic acid fermentation.

During the utilization of lignocellulosic biomass such as corn stover, many by-products are produced in the pretreatment process that can severely inhibit the activity of microbes in the fermentation step. To achieve efficient biomass conversion, detoxification is usually required before microbial fermentation. In this study, the prehydrolysate from dilute acid pretreatment of corn stover was used as a lactic acid fermentation substrate. Biochars made from corn stover (CSB), cow manure (CMB), and a mixture of corn stover and cow manure (MB) were applied for the detoxification of the prehydrolysate. All three types of biochar had a porous structure with a specific surface area ranging from 4.08 m2 g-1 (CMB) to 7.03 m2 g-1 (MB). After detoxification, both the numbers of inhibitors and their concentrations in the prehydrolysate decreased, indicating that the biochars prepared in this study were effective in inhibitor removal. The concentration of lactic acid obtained from the prehydrolysate without detoxification was only 12.43 g L-1 after fermentation for 96 h with a productivity of 0.13 g (L h)-1. Although the specific area of CMB was the lowest among the three biochars, the CMB-treated prehydrolysate resulted in the highest lactic acid concentration of 39.25 g L-1 at 96 h with a productivity of 0.41 g (L h)-1. The lactic acid bacteria in the CMB-treated prehydrolysate grew faster than the other two biochars, reaching an OD value of 8.12 at 48 h. The results showed promise for the use of agricultural wastes to make biochar to increase the yield of lactic acid fermentation through the detoxification process.

Polycationic Surfaces Promote Whole-Cell Immobilization and Induce Microgranulation of Clostridium saccharoperbutylacetonicum N1-4 for Enhanced Biobutanol Production.

Immobilization is a common strategy used to protect microbial cells to improve the performance of bioprocesses. However, the interaction mechanism between the cells and the immobilization material is generally poorly understood. In this study, we employed natural polysaccharide-based materials as immobilization carriers for clostridial fermentation in an attempt to enhance the production of butanol (a valuable biofuel/biochemical but highly toxic to the host cells) and meanwhile elucidate the interaction mechanisms related to immobilization. The utilization of chitosan powder as the immobilization carrier enhanced butanol productivity by 97% in the fermentation with Clostridium saccharoperbutylacetonicum N1-4 and improved butanol titer by 21% in the fermentation with Clostridium beijerinckii NCIMB 8052. Additionally, analogue derivatives using microcrystalline cellulose (MCC) and cotton cationized on the surface with 3-chloro-2-hydroxypropyltrymethylammonium (CHPTA) and 2-chloro-N,N-diethylaminoethyl chloride (DEAEC) were prepared and used as immobilization carriers for similar fermentation conditions. The CHPTA derivatives showed slightly increased production of butanol and total solvent with C. saccharoperbutylacetonicum. Overall, our results indicated that the interaction between the cell and the carrier material occurs through a double mechanism involving adsorption immobilization and induced aggregation. This work provides insights concerning the effects of the chemical properties of the carrier material (such as the cation density and surface area) on fermentation performance, enabling a better understanding of the interaction between bacterial cells and the cationic materials. The derivatization strategies employed in this study can be applied to most cellulosic materials to modulate the properties and enhance the interaction between the cell and the carrier material for immobilization, thus improving the bioprocess performance.

Spatial distribution and mass transport of Perfluoroalkyl Substances (PFAS) in surface water: A statewide evaluation of PFAS occurrence and fate in Alabama.

Per- and polyfluoroalkyl substances (PFAS) have been previously detected near suspected sources in Alabama, but the overall extent of contamination across the state is unknown. This study evaluated the spatial distribution of 17 PFAS within the ten major river basins in Alabama and provided insights into their transport and fate through a mass flux analysis. Six PFAS were identified in 65 out of the 74 riverine samples, with mean ∑6PFAS levels of 35.2 ng L-1. The highest ∑6PFAS concentration of 237 ng L-1 was detected in the Coosa River, a transboundary river that receives discharges from multiple sources in Alabama and Georgia. PFAS distribution was not observed to be uniform across the state: while the Coosa, Alabama, and Chattahoochee rivers presented relatively high mean ∑6PFAS concentrations of 191, 100 and 28.8 ng L-1, respectively, PFAS were not detected in the Conecuh, Escatawpa, and Yellow rivers. Remaining river systems presented mean ∑6PFAS concentrations between 7.94 and 24.7 ng L-1. Although the short-chain perfluoropentanoic acid (PFPeA) was the most detected analyte (88%), perfluorobutanesulfonic acid (PFBS) was the substance with the highest individual concentration of 79.4 ng L-1. Consistent increases in the mass fluxes of PFAS were observed as the rivers flowed through Alabama, reaching up to 63.3 mg s-1, indicating the presence of numerous sources across the state. Most of the mass inputs would not have been captured if only aqueous concentrations were evaluated, since concentration is usually heavily impacted by environmental conditions. Results of this study demonstrate that mass flux is a simple and powerful complementary approach that can be used to broadly understand trends in the transport and fate of PFAS in large river systems.

Substrate properties as controlling parameters in attached algal cultivation.

There is growing interest in attached algae cultivation systems because they could provide a more cost- and energy-efficient alternative to planktonic (suspended algae) cultivation systems for many applications. However, attached growth systems have been far less studied than planktonic systems and have largely emphasized algae strains of most interest for biofuels. New algal biorefinery pathways have assessed the commercial potentials of algal biomass beyond biofuel production and placed more emphasis on value-added products from that biomass. Therefore, algal strain selection criteria and biomass cultivation methods need to be updated to include additional strains for improved efficiency. One possible way of improving attached cultivation systems is through engineering substrate surface characteristics to boost algal adhesion and enable strain selective algal colonization and growth. This review explores the effect of substrate chemical and topographical characteristics on the cultivation of attached algae. It also highlights the importance of considering algal community structure and attachment mechanisms in investigating attached algae systems using the example of filamentous algae found in algal turf scrubber (ATS™) systems. KEY POINTS : • Attached algal cultivation is a promising alternative to planktonic cultivation. • Performance increase results from tuning surface qualities of attachment substrates. • Attachment adaptation of periphytic algae has innate potential for cultivation.

Identification and Investigation of Autolysin Genes in Clostridium saccharoperbutylacetonicum Strain N1-4 for Enhanced Biobutanol Production.

Biobutanol is a valuable biochemical and one of the most promising biofuels. Clostridium saccharoperbutylacetonicum N1-4 is a hyperbutanol-producing strain. However, its strong autolytic behavior leads to poor cell stability, especially during continuous fermentation, thus limiting the applicability of the strain for long-term and industrial-scale processes. In this study, we aimed to evaluate the role of autolysin genes within the C. saccharoperbutylacetonicum genome related to cell autolysis and further develop more stable strains for enhanced butanol production. First, putative autolysin-encoding genes were identified in the strain based on comparison of amino acid sequence with homologous genes in other strains. Then, by overexpressing all these putative autolysin genes individually and characterizing the corresponding recombinant strains, four key genes were pinpointed to be responsible for significant cell autolysis activities. Further, these key genes were deleted using CRISPR-Cas9. Fermentation characterization demonstrated enhanced performance of the resultant mutants. Results from this study reveal valuable insights concerning the role of autolysins for cell stability and solvent production, and they provide an essential reference for developing robust strains for enhanced biofuel and biochemical production.IMPORTANCE Severe autolytic behavior is a common issue in Clostridium and many other microorganisms. This study revealed the key genes responsible for the cell autolysis within Clostridium saccharoperbutylacetonicum, a prominent platform for biosolvent production from lignocellulosic materials. The knowledge generated in this study provides insights concerning cell autolysis in relevant microbial systems and gives essential references for enhancing strain stability through rational genome engineering.

Analysis of very-high surface area 3D-printed media in a moving bed biofilm reactor for wastewater treatment.

Moving Bed Biofilm Reactors (MBBRs) can efficiently treat wastewater by incorporating suspended biocarriers that provide attachment surfaces for active microorganisms. The performance of MBBRs for wastewater treatment is, among other factors, contingent upon the characteristics of the surface area of the biocarriers. Thus, novel biocarrier topology designs can potentially increase MBBR performance in a significant manner. The goal of this work is to assess the performance of 3-D-printed biofilter media biocarriers with varying surface area designs for use in nitrifying MBBRs for wastewater treatment. Mathematical models, rendering, and 3D printing were used to design and fabricate gyroid-shaped biocarriers with a high degree of complexity at three different levels of specific surface area (SSA), generally providing greater specific surface areas than currently available commercial designs. The biocarriers were inoculated with a nitrifying bacteria community, and tested in a series of batch reactors for ammonia conversion to nitrate, in three different experimental configurations: constant fill ratio, constant total surface area, and constant biocarrier media count. Results showed that large and medium SSA gyroid biocarriers delivered the best ammonia conversion performance of all designs, and significantly better than that of a standard commercial design. The percentage of ammonia nitrogen conversion at 8 hours for the best performing biocarrier design was: 99.33% (large SSA gyroid, constant fill ratio), 94.74% (medium SSA gyroid, constant total surface area), and 92.73% (large SSA gyroid, constant biocarrier media count). Additionally, it is shown that the ammonia conversion performance was correlated to the specific surface area of the biocarrier, with the greatest rates of ammonia conversion (99.33%) and nitrate production (2.7 mg/L) for manufactured gyroid biocarriers with a specific surface area greater than 1980.5 m2/m3. The results suggest that the performance of commercial MBBRs for wastewater treatment can be greatly improved by manipulation of media design through topology optimization.

Enhancing the tolerance of Clostridium saccharoperbutylacetonicum to lignocellulosic-biomass-derived inhibitors for efficient biobutanol production by overexpressing efflux pumps genes from Pseudomonas putida.

Furan aldehydes and phenolic compounds generated during biomass pretreatment can inhibit fermentation for biofuel production. Efflux pumps actively transport small molecules out of cells, thus sustaining normal microbial metabolism. Pseudomonas putida has outstanding tolerance to butanol and other small molecules, and we hypothesize that its efflux pump could play essential roles for such robustness. Here, we overexpressed efflux pump genes from P. putida to enhance tolerance of hyper-butanol producing Clostridium saccharoperbutylacetonicum to fermentation inhibitors. Interestingly, overexpression of the whole unit resulted in decreased tolerance, while overexpression of the subunit (srpB) alone exerted significant enhanced robustness of the strain. Compared to the control, the engineered strain had enhanced capability to grow in media containing 17% more furfural or 50% more ferulic acid, and produced ~14 g/L butanol (comparable to fermentation under regular conditions without inhibitors). This study provided valuable reference for boosting microbial robustness towards efficient biofuel production from lignocellulosic materials.

Engineering of bio-mimetic substratum topographies for enhanced early colonization of filamentous algae.

This work reveals a set of surface topography parameters that are significant for algal attachment to natural rock substrata. Topography analysis of rock surfaces from a stream identifies three descriptive areal parameters (Smr, Sv, and Sa) that correlate with the presence of natural periphyton community. A method was developed and validated to reverse engineer and manufacture artificial substrata with topographic complexity defined by these parameters, using computational modeling and additive manufacturing. Results from colonization experiments with filamentous algae show statistically significant increases in early biomass accrual rates on substrata with higher values of Sa and Sv parameters and lower values of Smr parameter. These results suggest that manipulation of the level of roughness (peak-to-valley distance and material ratio above the mean) and the distribution of hill and dale sequences can control initial colonization locations and biomass accrual rates, presumably by enhancing growth and recruitment of cells from the overlying flow into protected refugia spaces. As such, these findings provide an approach for optimizing the design of substratum for increased early biomass productivity for attached growth algae cultivation systems.

Utilization of solid catfish manure waste as carbon and nutrient source for lactic acid production.

The aim of this work was to study the solid waste (manure) produced by catfish as a potential feedstock for the production of lactic acid (LA) via fermentation. The solid waste contains high levels of both carbohydrates and nutrients that are sufficient for LA bacteria. Simultaneous saccharification and co-fermentation (SSCF) was applied using enzyme and Lactobacillus pentosus, and different loadings of enzyme and solid waste were tested. Results showed LA concentrations of 35.7 g/L were obtained at 15% solids content of catfish waste. Because of the high nutrient content in the fish waste, it could also be used as supplementary substrate for nitrogen and carbon sources with other lignocellulosic materials. A combined feedstock of catfish waste and paper mill sludge was tested, increasing the final LA concentration to 43.1 g/L at 12% solids loading. The catfish waste was shown to be a potential feedstock to provide both carbon and nutrients for LA production, suggesting its use as a sole substrate or in combination with other lignocellulosic materials.

Nutrient value of fish manure waste on lactic acid fermentation by Lactobacillus pentosus.

The aim of this work was to study the feasibility of using fish manure waste as a nutrient source for lactic acid fermentation. Fish waste contains nitrogen and minerals that could support the growth of lactic acid bacteria (LAB), making it a good candidate as the nutrient source for lactic acid fermentation. Two different fish manure wastes, from Nile tilapia and channel catfish aquaculture, were investigated for their performance on different sugar substrates. Both fish waste types showed low efficiency in the direct fermentation of glucose, but satisfactory efficiencies in simultaneous saccharification and fermentation (SSF) of cellulosic materials, such as pure cellulose and paper sludge. The highest lactic acid yield obtained was 87% and 91%, with a corresponding volumetric productivity of 1.006 and 0.580 g L-1 h-1, and corresponding lactic acid concentration of 96 and 56 g L-1 for cellulose and paper sludge, respectively. Fish waste concentrations did not show much impact on lactic acid production for the SSF process, where increasing fish waste from 10 to 30 g L-1 resulted in less than a 10% yield increase. In the present study, fish manure waste was shown to be an effective and economic nutrient source for lactic acid production by SSF.

Dynamic modeling predicts continued bioaccumulation of polybrominated diphenyl ethers (PBDEs) in smallmouth bass (Micropterus dolomiu) post phase-out due to invasive prey and shifts in predation.

Unprecedented food chain links between benthic and pelagic organisms are often thought to disrupt traditional contaminant transport and uptake due to changes in predation and mobilization of otherwise sequestered pollutants. A bioaccumulation model for polybrominated diphenyl ethers (PBDEs) is developed to simulate increases in biotic congener loads based upon trophic transfer through diet and gill uptake for a Lake Erie food chain including two invasive species as a benthic-pelagic link. The model utilizes species-specific bioenergetic parameters in a four-level food chain including the green alga Scenedesmus quadricauda, zebra mussels (Dreissena polymorpha), round goby (Appollonia melanostoma), and the smallmouth bass (Micropterus dolomiu). The model was calibrated to current biotic concentrations and predicts an increase in contaminant load by almost 48% in the upper trophic level in two years. Validation to archival data resulted in <2% error from reported values following a two-year simulation.

Association of nuisance filamentous algae Cladophora spp. with E. coli and Salmonella in public beach waters: impacts of UV protection on bacterial survival.

This study investigated whether filamentous algal species commonly found in nearshore public beach water systems provide protection from natural UV to bacteria present in the same environmental settings. To test this hypothesis, Cladophora spp., a filamentous nuisance algae group causing undesired water quality in the Great Lakes region was selected and its interactions with a non-pathogenic indicator organism Escherichia coli and a pathogenic strain of Salmonella enterica serovar Typhimurium were tested. In laboratory microcosms where the lake environment and natural sunlight conditions were simulated, a 7-log removal of E. coli was observed in only six hours of exposure to UV with an initial seed concentration of 10(3) CFU mL(-1). With the presence of algae, the same log removal was achieved in 16 hours. At higher seed concentrations of 10(5) CFU mL(-1), E. coli survived for two days with an extended survival up to 11 days in the presence of Cladophora spp. S. typhimurium has shown more resilient survival profiles, with the same log removals achieved in 14 and 20 days for low and high seed concentrations respectively, in the absence of algae. Cladophora spp. caused extended protection for S. typhimurium with much less log reductions reported. Algae-mediated protection from UV irradiation was attributed to certain organic carbon exuded from Cladophora spp. In addition, confocal microscopy images confirmed close interaction between bacteria and algae, more prominent with thin filamentous Cladophora spp.