Bo Fu

Acidogenic fermentation of proteinaceous sewage sludge: Effect of pH

Proteinaceous sewage sludge represents the sludge in which the protein content exceeds the carbohydrate content, accounts for a considerable part of sewage sludge. In this study, volatile fatty acids (VFA) production and synthesis pathway as well as bacterial community in acidogenic fermentation of proteinaceous sewage sludge under different pH conditions were investigated. The results indicated that the alkaline pH improved the solubilization and biodegradation of proteins in the sludge. The soluble protein concentration at pH 11.0 was 67.88% higher than that at pH 3.0. Its biodegradation efficiencies were reduced with a decrease in pH. The pH influenced not only the total VFA production yield, but also the percentage of individual VFA. The total VFA yield at pH 9.0 was 10.70 times of that at pH 3.0. The metabolic conversion pathway of proteins to VFA in acidogenic fermentation systems was investigated using a stoichiometry approach. The Stickland reaction was found to be the main pathway for the VFA accumulation from the decomposition of proteins. pH also significantly influenced the biodiversity and bacterial community in the system. The abundance of microorganisms under alkaline or acidic pH conditions was less than that under neutral pH condition, suggesting that most anaerobic fermentative microorganisms could not tolerate the hostile environment. The terminal restriction fragment length polymorphism analysis showed that, when the pH was reduced from 12.0 to 7.0 and finally to 3.0, the dominant bacterial genuses and percentage of the total microorganisms were Granulicatella genus (65%), Clostridium genus (28%) and Bacillus genus (71%), respectively.

Reactor performance and bacterial pathogen removal in response to sludge retention time in a mesophilic anaerobic digester treating sewage sludge

The effects of sludge retention time (SRT) on reactor performance and bacterial pathogen removal of sludge mesophilic anaerobic digestion (MAD) were investigated in a continuous stirred tank reactor. The average volatile solids removal remained around 20% and the biogas production rate varied from 100 to 132ml/ld. The MAD process was efficient to remove Salmonella sp. and Escherichia coli with removal efficiencies increased with SRT from 11d, 16d to 25d. However, the Shigella sp. removal was insignificant. The difference in the resistance of the three pathogens to sludge MAD process is helpful to the selection of pathogen indicators in the biosolids. Log reduction of pathogens determined by MPN was much higher than the data by quantitative PCR, suggesting the presence of viable but non-culturable pathogen cells. This study confirms that the control of appropriate SRT for sludge MAD should take both reactor performance and pathogen removal into account.

Improved volatile fatty acids anaerobic production from waste activated sludge by pH regulation: Alkaline or neutral pH?

In this study, the anaerobic fermentation was carried out for volatile fatty acids (VFAs) production at different pH (between 7.0 and 10.0) conditions with untreated sludge and heat-alkaline pretreated waste activated sludge. In the fermentation with untreated sludge, the extent of hydrolysis of organic matters and extent of acidification at alkaline pH are 54.37% and 30.37%, respectively, resulting in the highest VFAs yield at 235.46mg COD/gVS of three pH conditions. In the fermentation with heat-alkaline pretreated sludge, the acidification rate and VFAs yield at neutral pH are 30.98% and 240.14mg COD/gVS, respectively, which are higher than that at other pH conditions. With the glucose or bovine serum albumin as substrate for VFAs production, the neutral pH showed a higher VFAs concentration than the alkaline pH condition. The results of terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that the alkaline pH caused low microbial richness. Based on the results in this study, we demonstrated that the alkaline pH is favor of hydrolysis of organic matter in sludge while neutral pH improved the acidogenesis for the VFAs production from sludge. Our finding is obvious different to the previous research and helpful for the understanding of how heat-alkaline pretreatment and alkaline fermentation influence the VFAs production, and beneficial to the development of VFAs production process.

Quantification of viable but nonculturable bacterial pathogens in anaerobic digested sludge

Enteric bacterial pathogens in sewage sludge easily become viable but nonculturable (VBNC) during anaerobic digestion, which escape detection by standard culture methods and pose a potential health risk. In this study, a method that is combining the standard culture method with the reverse transcription quantitative PCR (RT-qPCR) assay was developed for the quantification of bacterial pathogens in the VBNC state. The cycle threshold (CT) values from RT-qPCR assays were linear to the bacterial number in the range from 109 to 102 most probable number (MPN) per reaction for Escherichia coli (R2 = 0.9964) and Salmonella typhimurium (R2 = 0.9938) and from 109 to 104 MPN per reaction for Shigella flexneri (R2 = 0.997), respectively. Mesophilic anaerobic digestion (MAD) caused the bacterial pathogens in sewage sludge entering into VBNC state with the incidence indexes of 0.01–1.12 for E. coli, 2.48–436.52 for S. typhimurium, and 4.17–6.61 for S. flexneri, respectively. Given different VBNC incidence indexes of bacterial pathogens in sewage sludge by MAD, the quantification results of VBNC pathogens using RT-qPCR could provide an improved evaluation of pathogen inactivation efficiency and biological safety in sludge anaerobic digestion.

Occurrence and reactivation of viable but non-culturable E. coli in sewage sludge after mesophilic and thermophilic anaerobic digestion

The occurrence and reactivation of viable but non-culturable (VBNC) Escherichia coli after different anaerobic digestions and the subsequent dewatering and storage were evaluated and compared. Culturable E. coli in digested sludge increased by two to four orders of magnitudes immediately after dewatering. However, counts of both the total and viable E. coli indicated that the increase of E. coli was attributed to its reactivation from the VBNC state to the culturable state. The VBNC pathogen incidences of thermophilic digestion were two to three orders of magnitude higher than those of mesophilic digestion. Accordingly, culturable E. coli in thermophilic, digested sludge after storage were one order of magnitude higher than mesophilic digestion. Anaerobic digestion thus mainly alters the culturable state of pathogens rather than killing them; therefore the biological safety of digested sludge, especially temperature-phased anaerobic digestion, should be carefully assessed.

Improving volatile fatty acid yield from sludge anaerobic fermentation through self-forming dynamic membrane separation.

Self-forming dynamic membrane (SFDM) separation was applied to the conventional sludge fermenter for improving VFA yields. Results indicated SFDM presented good performance in transferring products, retaining substrates, and enriching useful bacteria. The retention ratios of suspended solids, soluble COD, proteins, and polysaccharides reached 99%, 30%, 70%, and 40%, respectively, and more than 90% of the VFAs and ammonia could be transferred in a timely manner. The structure of the microbial community was optimized, which led to enhanced releases of hydrolytic enzymes and accelerated enrichments of functional bacteria. Protease and β-glucosidase activities increased from 1.0 to 5.0U/mL and 15.0 to 23.0μmol/L·h, respectively. VFA yield and sludge conversion ratio increased by 233.3% and 227.9%, respectively. Moreover, SFDM had good operation stability, including a short formation time, a long operation period, and a low transmembrane pressure. These results show VFA yield from sludge fermentation can be greatly improved by SFDM separation.

Improving volatile fatty acids production by exploiting the residual substrates in post-fermented sludge: Protease catalysis of refractory protein.

The real cause to the low yield of volatile fatty acids (VFAs), from inhibition or low biodegradation, is uncertain in sludge anaerobic fermentation. In this study, poor biodegradability of proteins and fast decrease of the indigenous hydrolase activity in the residual post-fermented sludge were found to be the major reasons. With the addition of trypsin or alkaline protease in residual post-fermented sludge after primary alkaline fermentation, degradation efficiency of refractory protein increased by 33.6% and 34.8%, respectively. Accordingly, the VFAs yields were improved by 69.7% and 106.1%, respectively. Furthermore, the activities of added trypsin and alkaline protease could maintain at 13.52 U/mL and 19.11 U/mL in the alkaline fermentation process. This study demonstrated that exploiting the refractory proteins in residual post-fermented sludge by protease addition seems to be a very promising way for improving VFAs yield of conventional alkaline fermentations with waste activated sludge.

Novel insight into the relationship between organic substrate composition and volatile fatty acids distribution in acidogenic co-fermentation

BackgroundCo-fermentation is an attractive technology for improving volatile fatty acids (VFAs) production by treatment of solid organic wastes. However, it remains unclear how the composition of different organic matters in solid waste influences the VFAs distribution, microbial community structure, and metabolic pathway during acidogenic co-fermentation. In this study, different organic wastes were added into waste activated sludge (WAS) as co-fermentation substrates to explore the impact of organic matter composition on VFAs pattern and the microbiological mechanism .ResultsAcetate was the most dominant VFA produced in all fermentation groups, making up 41.3–57.6% of the total VFAs produced during acidogenic co-fermentation under alkaline condition. With the increased addition of potato peel waste, the concentrations of propionate and valerate decreased dramatically, while ethanol and butyrate concentrations increased. The addition of food waste caused gradual decreases of valerate and propionate, but ethanol increased and butyrate was relatively stable. Some inconsistency was observed between hydrolysis efficiency and acidification efficiency. Our results revealed that starch was mainly responsible for butyrate and ethanol formation, while lipids and protein favored the synthesis of valerate and propionate. Microbial community analysis by high-throughput sequencing showed that Firmicutes had the highest relative abundance at phylum level in all fermentation groups. With 75% potato peel waste or 75% food waste addition to WAS, Bacilli (72.2%) and Clostridia (56.2%) were the dominant respective classes. In fermentation using only potato peel waste, the Bacilli content was 64.1%, while the Clostridia content was 53.6% in the food-only waste fermentation.ConclusionsAcetate was always the dominant product in acidogenic co-fermentation, regardless of the substrate composition. The addition of carbon-rich substrates significantly enhanced butyrate and ethanol accumulation, while protein-rich substrate substantially benefited propionate and valerate generation. Potato peel waste substantially favored the enrichment of Bacilli, while food waste dramatically increased Clostridia content in the sludge.

Quantification of viable but nonculturable Salmonella spp. and Shigella spp. during sludge anaerobic digestion and their reactivation during cake storage

The presence of viable but nonculturable (VBNC) bacterial pathogens which often fail to be detected by cultivation and can regain the cultivability if the living conditions improve were reported. The objective of this study was to determine the occurrence of VBNC Salmonella spp. and Shigella spp. in the biosolids during anaerobic digestion and its reactivation during the cake storage.

Control of C/N ratio for butyric acid production from textile wastewater sludge by anaerobic digestion

Increasing textile wastewaters and their biotreatment byproduct-waste activated sludge are serious pollution problems. Butyric acid production from textile wastewater sludge by anaerobic digestion at different C/N ratios was investigated. Adding starch to textile wastewater sludge with a C/N ratio of 30 increased the butyric acid concentration and percentage accounting for total volatile fatty acids (TVFAs) to 21.42 g/L and 81.5%, respectively, as compared with 21.42 g/L and 10.6% of textile wastewater sludge alone. The maximum butyric acid yield (0.45 g/g VS), conversion rate (0.74 g/g VS(digest)) and production rate (2.25 g/L d) was achieved at a C/N ratio of 30. The biological toxicity of textile wastewater sludge also significantly decreased after the anaerobic digestion. The study indicated that the anaerobic co-digestion of textile wastewater sludge and carbohydrate-rich waste with appropriate C/N ratio is possible for butyric acid production.