Nitrospira dominant pin-point flocs with granule-like settleability in stirred tank reactors with oxic/hypoxic/oxic zones

The characteristics of biomass and microbial community dynamics, in relation to autotrophic nitrification, were studied in two 20 L stirred tank reactors (STR) with oxic/hypoxic/oxic zones. The bioreactors were fed with synthetic wastewater with stepwise increasing ammonia concentrations (50–200 N mg/L) without organic substrate in the first phase (autotrophic phase) for 35 days (R1) and 15 days (R2), followed by a heterotrophic phase (with supplementation of organic substrate). The settling properties of the biomass, represented by pin-point flocs, gradually improved in both reactors during the autotrophic phase. The pin-point flocs of R1 exhibited granule-like settling properties. The SVI30 in RI gradually improved to 29 mL/g MLSS, and the corresponding SVI30/ SVI10 gradually improved to 0.88 during the autotrophic phase. The settling properties of the biomass deteriorated in both bioreactors during the heterotrophic phase. The protein to polysaccharide ratio (PN:PS ratio) gradually increased in the extracted EPS (in both, loosely bound (LB) and tightly bound (TB) EPS) during the autotrophic phase, in both bioreactors. The TB:LB EPS ratio was higher when the pin-point flocs of R1 showed granule-like settling properties, followed by a decline in TB:LB EPS ratio during the heterotrophic phase. A combination of molecular approaches (droplet digital-PCR (dd-PCR) and 16S rRNA gene sequencing) revealed that Nitrospira were the predominant nitrifying bacteria in the pin-point flocs that show granular sludge-like settling properties during autotrophic phase in R1. Comammox Nitrospira was the dominant ammonia oxidizer in seed biomass and at low ammonia concentrations in both bioreactors. The relative abundance of canonical ammonia-oxidizing bacteria increased with an increase in influent-ammonia concentrations

Form and Function of Clostridium Thermocellum Biofilms

The importance of bacterial adherence has been acknowledged in microbial lignocellulose conversion studies; however, few reports have described the function and structure of biofilms supported by cellulosic substrates. We investigated the organization, dynamic formation, and carbon flow associated with biofilms of the obligately anaerobic cellulolytic bacterium Clostridium thermocellum 27405. Using noninvasive, in situ fluorescence imaging, we showed biofilms capable of near complete substrate conversion with a characteristic monolayered cell structure without an extracellular polymeric matrix typically seen in biofilms. Cell division at the interface and terminal endospores appeared throughout all stages of biofilm growth. Using continuous-flow reactors with a rate of dilution (2 h1) 12-fold higher than the bacterium’s maximum growth rate, we compared biofilm activity under low (44 g/liter) and high (202 g/liter) initial cellulose loading. The average hydrolysis rate was over 3-fold higher in the latter case, while the proportions of oligomeric cellulose hydrolysis products lost from the biofilm were 13.7% and 29.1% of the total substrate carbon hydrolyzed, respectively. Fermentative catabolism was comparable between the two cellulose loadings, with ca. 4% of metabolized sugar carbon being utilized for cell production, while 75.4% and 66.7% of the two cellulose loadings, respectively, were converted to primary carbon metabolites (ethanol, acetic acid, lactic acid, carbon dioxide). However, there was a notable difference in the ethanol-to-acetic acid ratio (g/g), measured to be 0.91 for the low cellulose loading and 0.41 for the high cellulose loading. The results suggest that substrate availability for cell attachment rather than biofilm colonization rates govern the efficiency of cellulose conversion

Antibacterial Properties and Mechanism of Activity of a Novel Silver-Stabilized Hydrogen Peroxide.

Huwa-San peroxide (hydrogen peroxide; HSP) is a NSF Standard 60 (maximum 8 mg/L(-1)) new generation peroxide stabilized with ionic silver suitable for continuous disinfection of potable water. Experiments were undertaken to examine the mechanism of HSP against planktonic and biofilm cultures of indicator bacterial strains. Contact/kill time (CT) relationships that achieve effective control were explored to determine the potential utility in primary disinfection. Inhibitory assays were conducted using both nutrient rich media and a medium based on synthetic wastewater. Assays were compared for exposures to three disinfectants (HSP, laboratory grade hydrogen peroxide (HP) and sodium hypochlorite) at concentrations of 20 ppm (therefore at 2.5 and 5 times the NSF limit for HP and sodium hypochlorite, respectively) and at pH 7.0 and 8.5 in dechlorinated tap water. HSP was found to be more or equally effective as hypochlorite or HP. Results from CT assays comparing HSP and HP at different bacterial concentrations with neutralization of residual peroxide with catalase suggested that at a high bacterial concentration HSP, but not HP, was protected from catalase degradation possibly through sequestration by bacterial cells. Consistent with this hypothesis, at a low bacterial cell density residual HSP was more effectively neutralized as less HSP was associated with bacteria and therefore accessible to catalase. Silver in HSP may facilitate this association through electrostatic interactions at the cell surface. This was supported by experiments where the addition of mono (K(+)) and divalent (Ca(+2)) cations (0.005-0.05M) reduced the killing efficacy of HSP but not HP. Experiments designed to distinguish any inhibitory effect of silver from that of peroxide in HSP were carried out by monitoring the metabolic activity of established P. aeruginosa PAO1 biofilms. Concentrations of 70-500 ppm HSP had a pronounced effect on metabolic activity while the equivalent concentrations of ionic silver (50- 375 ppb) had a negligible effect, demonstrating that the microbiocidal activity of HSP was due to peroxide rather than silver. Overall, it was found that the antimicrobial activity of HSP is enhanced over that of hydrogen peroxide; the presence of the ionic silver enhances interactions of HSP with the bacterial cell surface rather than acting directly as a biocide at the tested concentrations

Toluene removal in an automated cyclical bioreactor

A control scheme was developed for the automation of toluene removal in a cyclical bioreactor. Toluene was added to the self-cycling fermentor by diffusion across a silicone membrane. Transient dissolved oxygen, carbon dioxide evolution, and oxidationreduction potential (ORP) were screened as potential control variables. Through experimentation, ORP was deemed most effective. Control algorithms based on realtime estimates of the first and second derivatives of the ORP signal were tested. Although both approaches resulted in stable operation of the reactor, average toluene removal efficiencies of 95% were realized when control was based on the second derivative. This was significantly higher than the 77% efficiencies obtained when the control scheme centered on the first derivative of the transient ORP signal. The system developed was self-regulating, ensuring that a high toluene removal rate, on the order of 1.1 g h -1 , was maintained from cycle to cycle

A Novel System for Real-Time, In Situ Monitoring of CO2 Sequestration in Photoautotrophic Biofilms

Climate change brought about by anthropogenic CO2 emissions has created a critical need for effective CO2 management solutions. Microalgae are well suited to contribute to efforts aimed at addressing this challenge, given their ability to rapidly sequester CO2 coupled with the commercial value of their biomass. Recently, microalgal biofilms have garnered significant attention over the more conventional suspended algal growth systems, since they allow for easier and cheaper biomass harvesting, among other key benefits. However, the path to cost-effectiveness and scaling up is hindered by a need for new tools and methodologies which can help evaluate, and in turn optimize, algal biofilm growth. Presented here is a novel system which facilitates the real-time in situ monitoring of algal biofilm CO2 sequestration. Utilizing a CO2-permeable membrane and a tube-within-a-tube design, the CO2 sequestration monitoring system (CSMS) was able to reliably detect slight changes in algal biofilm CO2 uptake brought about by light–dark cycling, light intensity shifts, and varying amounts of phototrophic biomass. This work presents an approach to advance our understanding of carbon flux in algal biofilms, and a base for potentially useful innovations to optimize, and eventually realize, algae biofilm-based CO2 sequestration

Impact of Hydrothermal Pretreatment Parameters on Mesophilic and Thermophilic Fermentation and Anaerobic Digestion of Municipal Sludge

Four parameters affecting hydrothermal pretreatment (HTP) of municipal sludge prior to anaerobic digestion and fermentation were investigated. Partial factorial design including several key HTP parameters at two distinct levels, including temperature (170 and 190 °C), retention time (RT) (10 and 30 min), pH (4 and 10), and solid content (SC) (4% and 16%), were studied. Further, the impact of HTP parameters on mesophilic and thermophilic fermentation was explored and compared. Results revealed a significant effect of all HTP parameters on COD solubilization, VFA, and methane yield. There were correlations between HTP parameters and process responses such as VFA yield and methane yield. HTP was found to increase COD solubilization and VFA production between 15 and 20% during thermophilic fermentation in relation to mesophilic treatment. All parameters, including SC, temperature, pH, and RT, were important contributing factors affecting methane production during anaerobic digestion. The highest methane production yield of 269 mL CH4/g TCOD added was observed at the highest SC (16%) and pH (10) and at the lower temperature (170 °C) and RT (10). HTP is expected to be combined with other intensification routes to treat waste with high solid contents improving the fermentation and anaerobic digestion processes