Enzyme enhanced lactic acid fermentation of swine manure and apple waste: Insights from organic matter transformation and functional bacteria.

Anaerobic co-fermentation is a favorable way to convert agricultural waste, such as swine manure (SM) and apple waste (AW), into lactic acid (LA) through microbial action. However, the limited hydrolysis of organic matter remains a main challenge in the anaerobic co-fermentation process. Therefore, this work aims to deeply understand the impact of cellulase (C) and protease (P) ratios on LA production during the anaerobic co-fermentation of SM with AW. Results showed that the combined use of cellulase and protease significantly improved the hydrolysis during the enzymatic pretreatment, thus enhancing the LA production in anaerobic acidification. The highest LA reached 41.02 ± 2.09 g/L within 12 days at the ratio of C/P = 1:3, which was approximately 1.26-fold of that in the control. After a C/P = 1:3 pretreatment, a significant SCOD release of 45.34 ± 2.87 g/L was achieved, which was 1.13 times the amount in the control. Moreover, improved LA production was also attributed to the release of large amounts of soluble carbohydrates and proteins with enzymatic pretreated SM and AW. The bacterial community analysis revealed that the hydrolytic bacteria Romboutsia and Clostridium_sensu_stricto_1 were enriched after enzyme pretreatment, and Lactobacillus was the dominant bacteria for LA production. This study provides an eco-friendly technology to enhance hydrolysis by enzymatic pretreatment and improve LA production during anaerobic fermentation.

Effects of self-produced lactic fermentation (SPLF) on GHG and VSC emissions during swine slurry storage.

Self-produced lactic fermentation (SPLF) is a new valued utilization technology, but its impact on gas emission remains unclear. The objective of this study is to investigate the effect of replacing the H2SO4 additive with SPLF on greenhouse gas (GHG), and volatile sulfur compound (VSC) emissions from swine slurry storage in a laboratory-scale study. In this study, SPLF is directed toward producing lactic acid (LA) through the anaerobic fermentation of slurry and apple waste under the most suitable conditions, with the LA concentration kept at 10,000-52000 mg COD/L and the pH remaining within 4.5 during the following 90 days of slurry storage. Compared with that in the slurry storage treatment (CK), the GHG emissions decreased by 86% and 87% in the SPLF and H2SO4 groups, respectively. The low pH (i.e., less than 4.5) inhibited the growth of Methanocorpusculum and Methanosarcina and resulted in very low mcrA gene copies in the SPLF group, leading to a reduction in CH4 emissions. The methanethiol, dimethyl sulfide, dimethyl disulfide, and H2S emissions in the SPLF group were reduced by 57%, 42%, 22%, and 87% and increased by 2206%, 61%, 173%, and 1856% in the H2SO4 group, respectively. Therefore, SPLF can be an innovative bioacidification technology for effectively reducing GHG and VSC emissions from animal slurry storage.

Revealing mechanism of micro-aeration for enhancing volatile fatty acids production from swine manure.

Micro-aeration is considered a new strategy for improving volatile fatty acids (VFAs) production of agricultural waste. This study investigated the effect and mechanism of micro-aeration of air and oxygen (O2) on VFAs production from swine manure. The results showed that Air-micro-aeration had the most significant improvement effect, with the highest VFAs of 8.21 g/L, which was increased by 22.4%. Moreover, the mixing effects of different micro-aeration were limited, and the microbial communities significantly varied. Firmicutes and Bacteroidota were the dominant hydrolytic and acidogenic bacteria, and Air-micro-aeration preferentially promoted electron transfer activity and energy generation. Methanosarcina, Methanocorpusculum, and Methanobrevibacter can adapt to environmental changes according to their different oxygen tolerance, and the consumption and conversion of VFAs by methanogens were slow under Air-micro-aeration condition. This study revealed mechanism of micro-aeration for improving VFAs production from swine manure, providing a theoretical basis for micro-aeration regulation optimization.

Antibiotic removal potential for low greenhouse gas emission process of anaerobic digestion (AD) producing volatile fatty acids (VFAs).

This study aimed to investigate the antibiotic of sulfachloropyridazine (SCP) reduction and its effects on volatile fatty acids (VFAs) accumulation and microbial community structures during the process of anaerobic digestion (AD) producing VFA. Results showed that initial SCP concentrations have a positive correlation with reduction of SCP and accumulation of VFAs. The removal rates of SCP were 22.21%, 30.00%, 39.31% and 42.59% and the maximum production of VFAs were 3947, 6180, 6462 and 6032 mg/L for initial SCP concentrations of 25, 50, 75 and 100 mg/kg·TS, respectively. SCP only altered bacterial composition by hastening growth of specific bacterial taxa, but didn't increase bacterial α-diversity.

Improving production of lactic acid and volatile fatty acids from dairy cattle manure and corn straw silage: Effects of mixing ratios and temperature.

Anaerobic co-fermentation (AcoF) of dairy cattle manure (DCM) and corn straw silage (CSS) for producing lactic acid (LA) and volatile fatty acids (VFAs) was investigated. Batch experiments were conducted at seven different DCM/CSS ratios and at mesophilic and thermophilic temperatures. Results indicated that the highest concentration of LA was 17.50 ± 0.70 g/L at DCM:CSS ratio of 1:3 and thermophilic temperature, while VFAs was 18.23 ± 2.45 g/L at mono-CSS fermentation and mesophilic temperature. High solubilization of thermophilic conditions contributed to LA accumulation in AcoF process. Presence of the CSS increased the relative abundance of Lactobacillus for LA production at thermophilic. Meanwhile, the abundance of Bifidobacterium was increased when CSS was added at mesophilic, which could conduce to VFAs production. This study provides a new route for enhancing the biotransformation of DCM and CSS into short-chain fatty acids, potentially bringing economic benefits to agricultural waste treatment.

Produce individual medium chain carboxylic acids (MCCA) from swine manure: Performance evaluation and economic analysis.

Environmental issues caused by untreated animal manure require the development of resource recovery from waste through a circular economy approach. Producing medium chain carboxylic acids (MCCA) with higher value than biogas from manure has become promising. The objective of this study was to develop an effective individual MCCA produce process utilizing manure. In this study, animal manure was firstly anaerobic fermentation into short chain fatty acids (SCFA), then acidified manure and ethanol were fed into the chain elongation reactor with gradually increasing the organic loading rate (OLR) from 7.0 to 18.5 gCOD/L/d, and the mixed MCCA was separated individually via a fractional distillation process. The SCFA fermentation occurred mainly at the first 10 days, and the optimum concentrations of SCFA for treatments at 2 %VS, 4 %VS and 6 %VS were 6.58, 10.40 and 14.10 g/L, respectively. For the chain elongation reactor, the maximum concentrations of n-caproate and n-caprylate were 10.25 and 0.63 g/L, respectively, which were comparable with that obtained from other complex wastes. Over 90% MCCA can be recovered from the fermentation broth via the optimized extractant of methyl tert-butyl ether (MTBE) and the fractional distillation system. Preliminary economic analysis shows that this MCCA production process presented a higher economic benefit (9.25 $/m3 manure) than traditional biogas production (2.65 $/m3 manure), making MCCA production from swine manure economically competitive. This work provides a new route for manure resource recovery besides the biogas process.

Roles of micro-aeration on enhancing volatile fatty acids and lactic acid production from agricultural wastes.

Micro-aeration was proven to be an environmentally friendly strategy for efficiently enhancing volatile fatty acids (VFAs) and lactic acid (LA) production. The roles of micro-aeration on mono-digestion of swine manure (SM) for VFAs production and co-digestion of SM with corn silage (CS) for LA production were investigated, respectively. In this study, micro-aeration increased the maximum VFAs concentration by 20.3% to 35.71 g COD/L, and shortened the time to reach the maximum from 18 days to 10 days. Micro-aeration limited the conversion of LA into VFAs, leading to LA accumulation effectively to be 26.08 g COD/L. Microbial community analysis suggested that Clostridium and Terrisporobacter were always the dominant bacteria with or without micro-aeration for VFAs production, but the relative abundance increased notably during the same period. However, Bifidobacterium, which could use the higher productivity metabolism pathway, i.e., Bifidum pathway to produce LA, increased from lower than 1% to 22.9% by micro-aeration.

Optimization of lactate production from co-fermentation of swine manure with apple waste and dynamics of microbial communities.

Co-anaerobic fermentation (co-AF) of swine manure (SM) and apple waste (AW) has been proved to be beneficial for lactic acid (LA) production. In order to further improve the LA production, three important parameters, namely AW in feedstock, temperature, volatile solids (VS) of feedstock, were evaluated using Box-Behnken design and response surface methodology. The quadratic regression model was developed and interactive effects was found between the three parameters. Results showed that the maximum concentration, 31.18 g LA/L (with LA yield of 0.62 g/g VS), was obtained under optimum conditions of 60.4% AW in feedstock, 34.7 ℃, and 5.0% VS. At the optimum conditions, the solubilization of organic matter was enhanced compared with mono-fermentation of SM. Microbial community structure of the reactor diverged greatly with fermentation time. Clostridium and Lactobacillus were dominant bacteria in the fermentation process, resulting in a remarkably LA accumulation.

Fate of antibiotics during membrane separation followed by physical-chemical treatment processes.

Membrane separation technology has been widely utilised to obtain clean permeate and concentrated nutrients from biogas digested slurry. However, some antibiotics are frequently found in digested slurry. Antibiotic removal during the steps, including paper filtration (PF), hollow-fibre membrane ultrafiltration (HF), nanofiltration (NF) and reverse osmosis (RO), of combined membrane separation processing (CMP) remain poorly understood. Experiments were performed on a pilot-scale CMP plant to investigate antibiotic transport and rejection efficiencies in CMP products and analyse the antibiotic removal performances of additional treatments. Results showed average relative antibiotic rejection efficiencies of 50%, 83%, 28% and 14% for PF, HF, NF and RO during CMP, respectively. Each CMP product needed additional treatment for antibiotic removal. Antibiotic removal capacity followed the order of fine halloysite > P25 > activated charcoal > coarse halloysite. The average antibiotic removal rates for different CMP products ranged from 73.3% to 99.9%. But antibiotics can only be adsorbed by fine halloysite and must be degraded through other treatments, Thus, P25 has considerable practical importance for the removal of antibiotics, as antibiotics can be degraded during photocatalysis.

Effect of Pretreatment on Hydraulic Performance of the Integrated Membrane Process for Concentrating Nutrient in Biogas Digestate from Swine Manure.

Nanofiltration (NF) or reverse osmosis (RO) process has been widely applied for concentrating nutrient in biogas digestate. However, efficient pretreatment is key to the sustainable operation of NF or RO. In this study, the combination of NF and RO for concentrating biogas digestate was compared using different pretreatments of hollow fiber ultrafiltration membrane (HFUFM) and ceramic membrane (CUFM). Pilot-scale batch tests were conducted (500 L). CUFM showed a higher membrane flux than HFUFM (100 ~ 180 L·(m2·h)-1 vs. 17 ~ 35 L·(m2·h)-1), but they showed little impact on the NF + RO process. Membrane fluxes of NF and RO were 20 ~ 48 L·(m2·h)-1 and 16 ~ 40 L·(m2·h)-1, respectively. In the RO permeates, the removal rates of total suspended solids (TSS), total solids (TS), chemical oxygen demand (COD), total nitrogen (TN), NH4+-N, and Cl- were above 91%. In the concentrates, TN and total potassium (TK) were concentrated by 1.60 and 2.00 folds in the NF stage, and by 2.10 and 2.30 folds in the RO stage. Further attention should be paid to the antibiotics risks in the concentrates before they are utilized as plant fertilizers.

Bioconversion of swine manure into high-value products of medium chain fatty acids.

This research proposes and demonstrates, for the first time, the utilization of swine manure as a complex feedstock to produce high-value medium chain fatty acids (MCFA). The two-stage anaerobic digestion (AD) carboxylates platform was adopted for the conversion of swine manure to short chain fatty acids (SCFAs) and then SCFAs to MCFA (n-caproate, n-heptanoate, and n-caprylate) with ethanol supplementation. We defined the appropriate initial pH of 10.0 for SCFAs production with a carbon conversion rate of 71.2%, and acetate, propionate were the main products, which accounted for around 72.9% of the total SCFAs in the primary stage (I). Through the addition of ethanol, 61.3% of the converted carbon in the complex SCFAs solution was converted into MCFA (C6-C8) in the chain elongation stage (II), while only 6.7% was attributed to methane formation. The concentrations of n-caproate, n-heptanoate, and n-caprylate reached 8.6 g COD/L (3.9 g/L), 6.4 g COD/L (2.7 g/L), and 2.6 g COD/L (1.07 g/L), respectively. This study achieved a relatively higher concentration of n-heptanoate compared with past studies of MCFA from other feedstock. These findings demonstrated a new route for resource recovery and the operating parameters for producing MCFA from swine manure.

Additional function of pasteurisation pretreatment in combination with anaerobic digestion on antibiotic removal.

Pasteurisation pretreatment (PP) in combination with anaerobic digestion (PPAD) is one of the best ways for sterilisation of pathogenic microorganisms in manure. However, the effect of antibiotic residues in manure on PPAD has not been studied. This study investigated the function of PPAD on antibiotic removal and the effect of antibiotic on PPAD performance. Results demonstrated that chlortetracycline (CTC) and oxytetracycline (OTC) concentrations decreased from 17.236 and 183.446 to 0 and 17.348 mg/kg·TS using PPAD, respectively. PPAD for swine manure containing CTC and OTC increased methane production from 244.0 ±â€¯7.6 to 254.0 ±â€¯6.1 mL/g·VS and reduced technical digestion time (T80) from 30 to 25 days compared with AD process. Moreover, PPAD affected only archaeal communities, whereas PP affected bacterial/archaeal communities. Thus, PPAD can be used to treat antibiotic-containing manure and reduce the negative effects of antibiotics.

The Combined Process of Paper Filtration and Ultrafiltration for the Pretreatment of the Biogas Slurry from Swine Manure.

The membrane process had been applied for the advanced treatment of pig farm biogas slurry. As studied, this physical pretreatment, with low cost and high efficiency of the suspended solids removal and nutrient retention, is required to control membrane fouling. The combined process of paper filtration and ultrafiltration in a pilot scale was applied in the present study. The main objective was to explore and identify the feasibility of the new process for the pretreatment of the separation liquid of pig farm biogas slurry. A precision identification experiment of paper filtration and the multi-batch repetitive experiments of the combined process were designed. The results showed, at the identified paper filtration precision of 50µm and an operating pressure of 0.3 bar of the ultrafiltration process, that the flux rate at the stable stage of the multi-batch operation was around 295.00 L/h. The combined process achieved an overall processing rate of 345.41 ± 18.81 L/h and a volume permeation proportion of 82.45% ± 0.85%. The TSS was removed by 95.71%, but total nitrogen (TN) and ammonia nitrogen (NH3-N) were retained by 76.29% ± 2.04% and 73.74% ± 2.10%, respectively. Comprehensively, the requirement for the pretreatment was obtained.

Effect of Time and Mixing in Thermal Pretreatment on Faecal Indicator Bacteria Inactivation.

Faecal indicator bacteria of faecal coliform, Salmonella spp., and faecal Streptococcus are present at high levels in faecal sludge and affect human health. Mesophilic anaerobic digestion cannot reduce faecal indicator bacteria to meet the standards for biosolids; therefore, the thermal pretreatment of faecal sludge is essential. The main objectives of this research were to evaluate the effect of thermal (70 °C) pretreatment time (20, 40, 60, 80, 100, and 120 min) and mixing velocity gradient (no mixing, 133, 191, and 238 s−1) on faecal indicator bacteria inactivation and determine the kinetics of the inactivation. The results showed that mixing has a more positive effect on pretreatment; thermal pretreatment with mixing was able to completely inactivate faecal indicator bacteria within 80 min, and inactivation followed first-order kinetics. In addition, under optimal mixing at a velocity gradient 191 s−1, the thermal pretreatment with mixing had a positive effect on the sludge solubilisation. Soluble chemical oxygen demand (SCOD, 71,430 mg L−1) and soluble protein (7.96 g L−1) were higher than the values obtained with thermal pretreatment without mixing, which were a SCOD value of 63,600 mg L−1 and soluble protein of 6.78 g L−1.

Antibiotic degradation and microbial community structures during acidification and methanogenesis of swine manure containing chlortetracycline or oxytetracycline.

Anaerobic digestion (AD) has been applied to animal manure stabilization, and antibiotics is frequently found in animal manure. However, antibiotic degradation and microbial community structures during two-stage AD (acidification and methanogenesis) remain poorly understood. This experiments on two-stage anaerobic swine manure digesters were performed to investigate the degradation mechanisms and effects of chlortetracycline (CTC) and oxytetracycline (OTC) on microbial community structures. Results showed that acidification and methanogenesis showed good degradation performance for manure containing CTC and OTC at 60 and 40 mg/kg·TS, respectively. CTC and OTC were degraded by 59.8% and 41.3% in the acidogenic stage and by 76.3% and 78.3% in the methanogenic stage, respectively. CTC and OTC negatively affected bacterial community in methanogenic and acidogenic stages, respectively. They also adversely influenced the archaeal species in the methanogenic stage. Two-stage AD was proposed to treat manure containing antibiotics and to reduce the negative effects of antibiotics on AD.

Effects of biofilter media depth and moisture content on removal of gases from a swine barn.

Media depth (MD) and moisture content (MC) are two important factors that greatly influence biofilter performance. The purpose of this study was to investigate the combined effect of MC and MD on removing ammonia (NH3), hydrogen sulfide (H2S), and nitrous oxide (N2O) from swine barns. Biofiltration performance of different MDs and MCs in combination based on a mixed medium of wood chips and compost was monitored. A 3 × 3 factorial design was adopted, which included three levels of the two factors (MC: 45%, 55%, and 67% [wet basis]; MD: 0.17, 0.33, and 0.50 m). Results indicated that high MC and MD could improve NH3 removal efficiency, but increase outlet N2O concentration. When MC was 67%, the average NH3 removal efficiency of three MDs (0.17, 0.33, and, 0.50 m) ranged from 77.4% to 78.7%; the range of average H2S removal efficiency dropped from 68.1-90.0% (1-34 days of the test period) to 36.8-63.7% (35-58 days of the test period); and the average outlet N2O concentration increased by 25.5-60.1%. When MC was 55%, the average removal efficiency of NH3, H2S, and N2O for treatment with 0.33 m MD was 72.8 ± 5.9%, 70.9 ± 13.3%, and -18.9 ± 8.1%, respectively; and the average removal efficiency of NH3, H2S, and N2O for treatment with 0.50 m MD was 77.7 ± 4.2%, 65.8 ± 13.7%, and -24.5 ±12.1%, respectively. When MC was 45%, the highest average NH3 reduction efficiency among three MDs was 60.7% for 0.5 m MD, and the average N2O removal efficiency for three MDs ranged from -18.8% to -12.7%. In addition, the pressure drop of 0.33 m MD was significantly lower than that of 0.50 m MD (p < 0.05). To obtain high mitigation of NH3 and H2S and avoid elevated emission of N2O and large pressure drop, 0.33 m MD at 55% MC is recommended. IMPLICATIONS: The performances of biofilters with three different media depths (0.17, 0.33, and 0.50 m) and three different media moisture contents (45%, 55%, and 67% [wet basis]) were compared to remove gases from a swine barn. Using wood chips and compost mixture as the biofilters media, the combination of 0.33 m media depth and 55% media moisture content is recommended to obtain good reduction of NH3 and H2S, and to simultaneously prevent elevated emission of N2O and large pressure drop across the media.

Effects of anaerobic digestion on chlortetracycline and oxytetracycline degradation efficiency for swine manure.

Manure containing antibiotics is considered a hazardous substance that poses a serious health risk to the environment and to human health. Anaerobic digestion (AD) could not only treatment animal waste but also generate valuable biogas. However, the interaction between antibiotics in manure and the AD process has not been clearly understood. In this study, experiments on biochemical methane potential (BMP) were conducted to determine the inhibition of the AD process from antibiotics and the threshold of complete antibiotic removal. The thresholds of the complete antibiotic removal were 60 and 40mg/kg·TS for CTC and OTC, respectively. CTC and OTC with concentrations below thresholds could increase the BMP of manure. When the CTC and OTC concentrations exceeded the thresholds, they inhibited manure fermentation, and the CTC removal rate declined exponentially with concentration (60-500mg/kg·TS). The relationship between OTC antibiotic concentration and its removal rate in AD treatment was described with exponential (40-100mg/kg·TS) and linear equations (100-500mg/kg·TS).

Study on anaerobic digestion treatment of hazardous colistin sulphate contained pharmaceutical sludge.

Pharmaceutical sludge is considered as a hazardous substance with high treatment and disposal fees. Anaerobic digestion could not only transform the hazardous substance into activated sludge, but also generate valuable biogas. This research had two objectives. First: studying the feasibility of anaerobic digestion and determining the biochemical methane potential (BMP) of pharmaceutical sludge under different Inoculum to substrate TS ratios (ISRs) of 0, 0.65, 2.58 and 10.32 in mesophilic condition of 37±1°C. Secondly, investigating the removal efficiency of colistin sulphate during anaerobic digestion. The results showed that the use of anaerobic digestion to treat the pharmaceutical sludge is feasible and that it can completely eliminate the colistin sulphate. The highest biogas production from pharmaceutical sludge is 499.46 mL/g TS at an ISR of 10.32.

Study on anaerobic treatment of hazardous steel-mill waste rolling oil (SmWRO) for multi-benefit disposal route.

Steel-mill waste rolling oil (SmWRO) is considered as hazardous substance with high treatment and disposal fees. Anaerobic process could not only transform the hazardous substance into activated sludge, but also generate valuable biogas. This study aimed at studying the biochemical methane potential of SmWRO under inoculum to substrate VS ratios (ISRs) of 0.25, 0.5, 1, 1.5, 2 and 3 using septic tank sludge as inoculum in mesophilic and thermophilic conditions, with blank tests for control. Specific biogas yield (mL/g VS(added)), net biogas yield (mL/g VS(removed)) and VS removal were analyzed. The ANOVA results indicated great influence of ISR and temperature on studied parameters. ISR of 1.5 at 55°C and ISR of 1.5 and 2 at 35°C were suggested with the highest specific biogas yield (262-265 and 303mL/g VS(added)). Kinetic analysis showed that Gompertz model fit the experimental data best with the least RMSE and largest R(2).