Single-dose rAAV5-based vaccine provides long-term protective immunity against SARS-CoV-2 and its variants.

The COVID-19 pandemic, caused by the SARS-CoV-2 virus and its variants, has posed unprecedented challenges worldwide. Existing vaccines have limited effectiveness against SARS-CoV-2 variants. Therefore, novel vaccines to match mutated viral lineages by providing long-term protective immunity are urgently needed. We designed a recombinant adeno-associated virus 5 (rAAV5)-based vaccine (rAAV-COVID-19) by using the SARS-CoV-2 spike protein receptor binding domain (RBD-plus) sequence with both single-stranded (ssAAV5) and self-complementary (scAAV5) delivery vectors and found that it provides excellent protection from SARS-CoV-2 infection. A single-dose vaccination in mice induced a robust immune response; induced neutralizing antibody (NA) titers were maintained at a peak level of over 1:1024 more than a year post-injection and were accompanied by functional T-cell responses. Importantly, both ssAAV- and scAAV-based RBD-plus vaccines produced high levels of serum NAs against the circulating SARS-CoV-2 variants, including Alpha, Beta, Gamma and Delta. A SARS-CoV-2 virus challenge showed that the ssAAV5-RBD-plus vaccine protected both young and old mice from SARS-CoV-2 infection in the upper and lower respiratory tracts. Whole genome sequencing demonstrated that AAV vector DNA sequences were not found in the genomes of vaccinated mice one year after vaccination, demonstrating vaccine safety. These results suggest that the rAAV5-based vaccine is safe and effective against SARS-CoV-2 and several variants as it provides long-term protective immunity. This novel vaccine has a significant potential for development into a human prophylactic vaccination to help end the global pandemic.

Integration of biological kinetics and computational fluid dynamics to model the growth of Nannochloropsis salina in an open channel raceway.

Microalgal growth and systemic productivity is not only affected by environmental conditions such as temperature, irradiance, and nutrient concentrations, but also by physical processes such as fluid flow and particulate sedimentation. Modeling and simulating the system is a cost-effective way to predict the growth behavior under various environmental and physical conditions while determining effective engineering approaches to maximize productivity. Many mathematical models have been proposed to describe microalgal growth, while computational fluid dynamics (CFD) have been used to model the behavior of many fluid systems. Integrating the growth kinetics into a CFD model can help researchers understand the impact of a variety of parameters and determine what measures can be taken to overcome some obstacles in the aquaculture industry--self-shading, biomass sedimentation, and contamination--which prevent the production of high biomass yields. The aim of this study was to integrate physical and environmental effects to predict space- and time-dependent algal growth in industrial scale raceways. A commercial CFD software, ANSYS-Fluent 14.5, was used to solve the proposed models in regards to fluid flow, heat transfer, and nutrient balance. User-defined functions written in C language were used to incorporate the kinetic equations into a three-dimensional standard k-ε turbulence model of an open channel raceway system driven by a single paddlewheel. Simulated results were compared with light intensity, temperature, nutrient concentration, and algal biomass data acquired for 56 day from an industrial scale raceway pond constructed for the growth of Nannochloropsis salina and were observed to be in good agreement with one another. There was up to a 17.6% increase in simulated productivity when the incoming CO2 concentration was increased from 0.0006 to 0.150 g L(-1), while the effect of paddlewheel velocity was not significant. Sensitivity analysis showed that the model was particularly sensitive to the species-specific maximum growth rate, light attenuation coefficient, optimal growth temperature, half-saturation constant for growth based on irradiance, and death coefficient.

Comparison of the microbial communities in solid-state anaerobic digestion (SS-AD) reactors operated at mesophilic and thermophilic temperatures.

The microbiomes involved in liquid anaerobic digestion process have been investigated extensively, but the microbiomes underpinning solid-state anaerobic digestion (SS-AD) are poorly understood. In this study, microbiome composition and temporal succession in batch SS-AD reactors, operated at mesophilic or thermophilic temperatures, were investigated using Illumina sequencing of 16S rRNA gene amplicons. A greater microbial richness and evenness were found in the mesophilic than in the thermophilic SS-AD reactors. Firmicutes accounted for 60 and 82 % of the total Bacteria in the mesophilic and in the thermophilic SS-AD reactors, respectively. The genus Methanothermobacter dominated the Archaea in the thermophilic SS-AD reactors, while Methanoculleus predominated in the mesophilic SS-AD reactors. Interestingly, the data suggest syntrophic acetate oxidation coupled with hydrogenotrophic methanogenesis as an important pathway for biogas production during the thermophilic SS-AD. Canonical correspondence analysis (CCA) showed that temperature was the most influential factor in shaping the microbiomes in the SS-AD reactors. Thermotogae showed strong positive correlation with operation temperature, while Fibrobacteres, Lentisphaerae, Spirochaetes, and Tenericutes were positively correlated with daily biogas yield. This study provided new insight into the microbiome that drives SS-AD process, and the findings may help advance understanding of the microbiome in SS-AD reactors and the design and operation of SS-AD systems.

Competitive partitioning of denitrification pathways during arrested methanogenesis: Implications in ammonium recovery, N2O emission, and volatile fatty acid production.

The complex interaction between nitrate (NO3-) reduction and fermentation is poorly understood when high levels of NO3- are introduced into anaerobic systems. This study investigated the competitive distribution between conventional denitrification (DEN) and dissimilatory nitrate reduction to ammonium (DNRA) during simultaneous denitrification and fermentation in arrested methanogenesis. Up to 62% of initial NO3- (200 mg-N/L) was retained as ammonium through DNRA at a chemical oxygen demand (COD)/N ratio of 25. Significant N2O emission occurred (1.7 - 8.0% of the initial NO3-) with limited carbon supply (≤1600 mg COD/L) and sludge concentration (≤3000 mg COD/L). VFA composition shifted predominantly towards acetic acid (>50%) in the presence of nitrate. A novel kinetic model was developed to predict DNRA vs. DEN partitioning and NO2- accumulation. Overall, NO3- input, organic loading, and carbon source characteristics independently and collectively controlled competitive DNRA vs. DEN partitioning.

Effect of alkaline pretreatment on photo-fermentative hydrogen production from giant reed: Comparison of NaOH and Ca(OH)2.

Giant reed was the first time used for photo-fermentative hydrogen production with HAU-M1 bacteria. Effects of NaOH and Ca(OH)2 pretreatments of giant reed on structural changes, enzymatic digestibility, hydrogen production, and energy conversion efficiency were evaluated. Compared to Ca(OH)2 pretreatment, NaOH pretreatment removed more dry matter and lignin at the same loading. The highest glucose yield (44.9%) of NaOH pretreatment was 1.74-fold higher than that of Ca(OH)2 pretreatment. 20% NaOH pretreated giant reed biomass achieved the highest hydrogen yield (98.3 mL/g TS), which was 20% and 70% higher than the highest level of Ca(OH)2 pretreated (20% Ca(OH)2) and untreated giant reed, respectively. Only giant reed biomass pretreated with 20% NaOH resulted in a significant (p < 0.05) increase (25%) in energy conversion efficiency.

Techno-economic analyses of solid-state anaerobic digestion and composting of yard trimmings.

Solid-state anaerobic digestion (SS-AD) and composting are two potential alternatives to divert yard trimmings from landfills. This study aimed to evaluate the techno-economic feasibility of commercial-scale SS-AD and composting systems (20,000 metric tons (MT)/year) that received both yard trimmings and liquid AD effluent using a modeling software, SuperPro Designer. Both the SS-AD and composting systems were shown to be economically feasible. While their revenues were comparable ($48/MT), SS-AD with digestate drying showed a higher capital cost ($256/MT vs. $84/MT) but a lower non-facility-dependent operating cost ($11/MT vs. $21/MT) than composting. The payback time, internal rate of return (IRR), and net present value (NPV) were estimated to be ∼10 years, 8%, and $0.2 million, respectively, for SS-AD, and ∼4.9 years, 33%, and $1.8 million, respectively, for composting. Digestate drying was necessary to make SS-AD profitable via the sale of byproduct, but it was also the most energy intensive step, relying on heat recovery to reduce costs. Moreover, the economics of SS-AD were highly improved (NPV $2 million) with financial incentives (i.e. investment tax credits), indicating that incentives were critical to the economic feasibility of current SS-AD systems that utilize lignocellulosic biomass. However, renewable identification numbers (RINs) and renewable energy certificates (RECs) had minor effects. Furthermore, the economics of both systems were most sensitive to plant size, tipping fees, and byproduct/compost price. The results suggest SS-AD may be favored for centralized management while composting for de-centralized management of yard trimmings. Alternative ways to valorize digestate should be evaluated in future studies.

Anaerobic digestion of food waste - Challenges and opportunities.

The disposal of large amounts of food waste has caused significant environmental pollution and financial costs globally. Compared with traditional disposal methods (i.e., landfilling, incineration, and composting), anaerobic digestion (AD) is a promising technology for food waste management, but has not yet been fully applied due to a few technical and social challenges. This paper summarizes the quantity, composition, and methane potential of various types of food waste. Recent research on different strategies to enhance AD of food waste, including co-digestion, addition of micronutrients, control of foaming, and process design, is discussed. It is envisaged that AD of food waste could be combined with an existing AD facility or be integrated with the production of value-added products to reduce costs and increase revenue. Further understanding of the fundamental biological and physicochemical processes in AD is required to improve the technology.

Sequential batch thermophilic solid-state anaerobic digestion of lignocellulosic biomass via recirculating digestate as inoculum - Part I: Reactor performance.

Sequential batch thermophilic (55°C) solid-state anaerobic digestion (SS-AD) of yard trimmings was evaluated at a total solids (TS) content of 22% via recirculating digestate as the inoculum. The substrate-to-inoculum (S/I) ratio of 1 (TS basis) was favored over 2 and 3 due to significantly higher methane yield and volumetric productivity. At an S/I ratio of 1, sequential batch SS-AD gradually reached steady state by 3 runs (30days/run) with increases in both methane yields (up to 11.5%) and cellulose degradation (up to 55%), indicating that recirculated digestate could be a feasible inoculum to establish long term stable SS-AD of lignocellulosic biomass. The initial sharp increases of volatile fatty acids during runs 2-4 indicated faster hydrolysis of organic matter than during run 1, suggesting that microbes were probably more acclimated due to digestate recirculation. At steady state, 51% (w/w) of the digestate was recirculated as the inoculum.

Sequential batch thermophilic solid-state anaerobic digestion of lignocellulosic biomass via recirculating digestate as inoculum - Part II: Microbial diversity and succession.

This study aimed to investigate the effect of recirculation of digestate as inoculum on the microbial communities in thermophilic solid-state anaerobic digestion (SS-AD) of yard trimmings. The SS-AD consisted of 4 consecutive runs (30days/run), with digestate from the previous run being used as the inoculum of the subsequent run. Bacterial and archaeal communities (day 0, 4, 8, 12, 20, and 30) were examined using Illumina sequencing of 16S rRNA genes. The results revealed substantial microbial succession toward increased diversity until run 3. The proportions of Firmicutes that contained cellulolytic bacteria doubled, which might explain the concomitantly increased cellulose degradation and volatile fatty acids (VFAs). Clostridia and Thermotogae appeared to correlate with VFAs. The VFA accumulation likely induced dynamic shifts of methanogens, particularly to hydrogenotrophic Methanothermobacter, implying that non-acetoclastic oxidative pathway dominated during the steady-state thermophilic SS-AD. This study suggested that recirculating SS-AD digestate might be an effective way for inoculation.

Draft Genome Sequence of Methylocaldum sp. SAD2, a Methanotrophic Strain That Can Convert Raw Biogas to Methanol in the Presence of Hydrogen Sulfide.

The draft genome sequence of Methylocaldum sp. SAD2, a methanotrophic strain isolated from a hydrogen sulfide-rich anaerobic digester, is reported here. Strain SAD2 possesses genes for methane oxidation in the presence of H2S.

Draft Genome Sequence of Methylocaldum sp. Strain 14B, an Obligate Hydrogen Sulfide-Tolerant Methanotrophic Strain That Can Convert Biogas to Methanol.

The draft genome sequence of Methylocaldum sp. 14B, an obligate methanotrophic strain isolated from solid-state anaerobic digestion systems, is reported here. Strain 14B possesses genes for methane oxidation and exhibited tolerance to H2S.

Comparison of sodium hydroxide and calcium hydroxide pretreatments of giant reed for enhanced enzymatic digestibility and methane production.

NaOH pretreatment with leachate reuse and Ca(OH)2 pretreatment were compared for improved enzymatic digestibility and biogas production from giant reed, a promising energy crop. The NaOH pretreatment with leachate reuse increased glucose yields during enzymatic hydrolysis by 2.6-fold, and methane yields during anaerobic digestion by 1.4- to 1.6-fold. However, NaOH pretreatment had a negative net benefit (i.e., revenue from increased energy production minus chemical cost). Pretreatment with 7-20% Ca(OH)2 not only improved glucose yield and methane yield by up to 2.3-fold and 1.4-fold, respectively, but also obtained a net benefit of $1.1-5.8/tonne dry biomass. Thus, Ca(OH)2 pretreatment was shown to be more feasible than NaOH pretreatment for biogas production from giant reed.

Availability of crop residues as sustainable feedstock for bioethanol production in North Carolina.

The amount of corn stover and wheat straw that can be sustainably collected in North Carolina was estimated to be 0.64 and 0.16 million dry t/yr, respectively. More than 80% of these crop residues are located in the coastal area. The bioethanol potential from corn stover and wheat straw was estimated to be about 238 million L (63 million gal/yr) in North Carolina. The future location of ethanol plant in North Carolina was estimated based on feedstock demand and collection radius. It is possible to have four ethanol plants with feedstock demand of 400, 450, 500, and 640 dry t/d. The collection radii for these four ethanol plants are 46, 60, 42, and 67 km (28, 37, 26, and 42 miles), respectively. The best location for a bioethanol plant includes four counties (Beaufort, Hyde, Tyrrell, and Washington) with feedstock demand of 500 t/d and collection radius about 26 mile.

Lactic acid recovery from cheese whey fermentation broth using combined ultrafiltration and nanofiltration membranes.

The separation of lactic acid from lactose in the ultrafiltration permeate of cheese whey broth was studied using a cross-flow nanofiltration membrane unit. Experiments to test lactic acid recovery were conducted at three levels of pressure (1.4, 2.1, and 2.8 MPa), two levels of initial lactic acid concentration (18.6 and 27 g/L), and two types of nanofiltration membranes (DS-5DK and DS-5HL). Higher pressure caused significantly higher permeate flux and higher lactose and lactic acid retention (p < 0.0001). Higher initial lactic acid concentrations also caused significantly higher permeate flux, but significantly lower lactose and lactic acid retention (p < 0.0001). The two tested membranes demonstrated significant differences on the permeate flux and lactose and lactic acid retention. Membrane DS-5DK was found to retain 100% of lactose at an initial lactic acid concentration of 18.6 g/L for all the tested pressures, and had a retention level of 99.5% of lactose at initial lactic acid concentration of 27 g/L when the pressure reached 2.8 MPa. For all the tests when lactose retention reached 99-100%, as much as 64% of the lactic acid could be recovered in the permeate.

Solid-state anaerobic digestion of lignocellulosic biomass: Recent progress and perspectives.

Solid-state anaerobic digestion (SS-AD), which has gained popularity in the past decade as an environmentally friendly and cost-effective technology for extracting energy from various types of lignocellulosic biomass, is reviewed in this paper. According to data of biomass and methane yields of lignocellulosic feedstocks, crop residues have the highest methane production potential in the U.S., followed by the organic fraction of municipal solid waste (OFMSW), forestry waste, and energy crops. Methane yield and process stability of SS-AD can be improved by different strategies, such as co-digestion with other organic wastes, pretreatment of lignocellulosic biomass, and optimization of operating parameters. Different models for SS-AD have been developed, and insights into SS-AD processes have been obtained via microbial community analysis, microscope imaging, and tracer techniques. Future research and development in SS-AD, including feedstock identification and co-digestion, feedstock storage and pretreatment, SS-AD reactor development, digestate treatment, and value-added production, are recommended.

Recovery of failed solid-state anaerobic digesters.

This study examined the performance of three methods for recovering failed solid-state anaerobic digesters. The 9-L digesters, which were fed with corn stover, failed at a feedstock/inoculum (F/I) ratio of 10 with negligible methane yields. To recover the systems, inoculum was added to bring the F/I ratio to 4. Inoculum was either added to the top of a failed digester, injected into it, or well-mixed with the existing feedstock. Digesters using top-addition and injection methods quickly resumed and achieved peak yields in 10days, while digesters using well-mixed method recovered slowly but showed 50% higher peak yields. Overall, these methods recovered 30-40% methane from failed digesters. The well-mixed method showed the highest methane yield, followed by the injection and top-addition methods. Recovered digesters outperformed digesters had a constant F/I ratio of 4. Slow mass transfer and slow growth of microbes were believed to be the major limiting factors for recovery.

Comparison between ensilage and fungal pretreatment for storage of giant reed and subsequent methane production.

Ensilage and fungal pretreatment of giant reed harvested from August through December were compared based on their effects on feedstock preservation, glucose yield, and subsequent methane production via anaerobic digestion (AD). Compared to fungal pretreatment, ensilage obtained lower total solids (<1.2%) and cellulose (<3.5%) losses, and comparable hemicellulose degradation, except for giant reed harvested in August. Ensilage increased glucose and methane yields by 7-15% and 4-14%, respectively, for giant reed harvested from August through December. Fungal pretreatment failed for giant reed harvested in August and October with reduced glucose yields, and was effective for that harvested in November and December, with about 20% increases in glucose yield. However, hydrocarbon losses during fungal pretreatment offset the increased glucose yield, resulting in decreased methane yields by AD. In summary, ensilage was found to be more suitable than fungal pretreatment for giant reed storage and its methane production via AD.

Value-added processing of crude glycerol into chemicals and polymers.

Crude glycerol is a low-value byproduct which is primarily obtained from the biodiesel production process. Its composition is significantly different from that of pure glycerol. Crude glycerol usually contains various impurities, such as water, methanol, soap, fatty acids, and fatty acid methyl esters. Considerable efforts have been devoted to finding applications for converting crude glycerol into high-value products, such as biofuels, chemicals, polymers, and animal feed, to improve the economic viability of the biodiesel industry and overcome environmental challenges associated with crude glycerol disposal. This article reviews recent advances of biological and chemical technologies for value-added processing of crude glycerol into chemicals and polymers, and provides strategies for addressing production challenges.

Value-added conversion of waste cooking oil and post-consumer PET bottles into biodiesel and polyurethane foams.

A sustainable process of value-added utilization of wastes including waste cooking oil (WCO) and post-consumer PET bottles for the production of biodiesel and polyurethane (PU) foams was developed. WCO collected from campus cafeteria was firstly converted into biodiesel, which can be used as vehicle fuel. Then crude glycerol (CG), a byproduct of the above biodiesel process, was incorporated into the glycolysis process of post-consumer PET bottles collected from campus to produce polyols. Thirdly, PU foams were synthesized through the reaction of the above produced polyols with isocyanate in the presence of catalysts and other additives. The characterization of the produced biodiesel demonstrated that its properties meet the specification of biodiesel standard. The effect of crude glycerol loading on the properties of polyols and PU foams were investigated. All the polyols showed satisfactory properties for the production of rigid PU foams which had performance comparable to those of some petroleum-based analogs. A mass balance and a cost analysis for the conversion of WCO and waste PET into biodiesel and PU foams were also discussed. This study demonstrated the potential of WCO and PET waste for the production of value-added products.

Comparison of digestate from solid anaerobic digesters and dewatered effluent from liquid anaerobic digesters as inocula for solid state anaerobic digestion of yard trimmings.

To select a proper inoculum for the solid state anaerobic digestion (SS-AD) of yard trimmings, digestate from solid anaerobic digesters and dewatered effluent from liquid anaerobic digesters were compared at substrate-to-inoculum (S/I) ratios from 0.2 to 2 (dry basis), and total solids (TS) contents from 20% to 35%. The highest methane yield of around 244L/kg VSfeed was obtained at an S/I ratio of 0.2 and TS content of 20% for both types of inoculum. The highest volumetric methane productivity was obtained with dewatered effluent at an S/I ratio of 0.6 and TS content of 24%. The two types of inoculum were found comparable regarding methane yields and volumetric methane productivities at each S/I ratio, while using dewatered effluent as inoculum reduced the startup time. An S/I ratio of 1 was determined to be a critical level and should be set as the upper limit for mesophilic SS-AD of yard trimmings.