Aerobic granular sludge for high-strength ammonium wastewater treatment: Effect of COD/N ratios, long-term stability and nitrogen removal pathways.

Aerobic granular sludge (AGS) technology is increasingly considered for wastewater treatment. AGS stability particularly under lower COD/N ratios is an impediment for AGS technology. This study evaluated AGS stability and nitrogen removal at different loading rates of 0.03 to 4 kg NH4+-N m-3 d-1 and COD/N ratios of 18.3 to 0.13. Ammoniacal and total nitrogen removals were high at 99.9% and 99.3%, respectively, during 440 days. MiSeq sequencing revealed a reduction in bacterial diversity and enrichment of ammonia oxidizing bacteria (AOB), anammox and denitrifying bacteria. Quantitative PCR showed enrichment of AOB, anammox bacteria, Nitrospira and denitrifiers. Chemical data and bacterial community supported occurrence of nitritation and anammox pathways. AGS had stable granular structure with excellent settling properties at lower COD/N ≤ 1. Removal of high-strength ammonium could be partly explained by the existing nitrogen pathways suggesting novel mechanisms. Nevertheless, results presented here support implementation of AGS process for ammonium wastewaters.

Sustainable bioreduction of toxic levels of chromate in a denitrifying granular sludge reactor.

Biological removal of chromate [Cr(VI)] in the presence or absence of nitrate by granular sludge biofilms was investigated in batch experiments and in a sequencing batch reactor (SBR). Denitrifying granular sludge cultivated from activated sludge was able to directly reduce Cr(VI) in the presence of an electron donor. Bioreduction was dependent on the initial Cr(VI) and the granular sludge concentrations. Bioreduction of Cr(VI) was followed by Cr(III) precipitation or entrapment in the granular sludge which was corroborated with decrease in total soluble Cr and increase in inorganic content of biomass. Batch experiments revealed that Cr(VI) addition has no major influence on high-strength nitrate (3000 mg L-1) denitrification, but nitrite denitrification was slowed-down. However, SBR experiment demonstrated successful denitrification as well as Cr(VI) removal due to enrichment of Cr(VI)-tolerant denitrifying bacteria. In fact, stable SBR performance in terms of complete and sustained removal of 0.05, 0.1, 0.2, 0.3, 0.5 and 0.75 mM Cr(VI) and denitrification of 3000 mg L-1 was observed during 2 months of operation. Active biomass and electron donor-dependent Cr(VI) removal, detection of Cr(III) in the biomass and recovery of ~ 92% of the Cr from the granular sludge biofilms confirms bioreduction followed by precipitation or entrapment of Cr(III) as the principal chromate removal mechanism. Metagenomic bacterial community analysis showed enrichment of Halomonas sp. in denitrifying granular sludge performing either denitrification or simultaneous reduction of nitrate and chromate.

Aerobic granular sludge technology: Mechanisms of granulation and biotechnological applications.

Aerobic granular sludge (AGS) is a novel microbial community which allows simultaneous removal of carbon, nitrogen, phosphorus and other pollutants in a single sludge system. AGS is distinct from activated sludge in physical, chemical and microbiological properties and offers compact and cost-effective treatment for removing oxidized and reduced contaminants from wastewater. AGS sequencing batch reactors have shown their utility in the treatment of abattoir, live-stock, rubber, landfill leachate, dairy, brewery, textile and other effluents. AGS is extensively researched for wide-spread implementation in sewage treatment plants. However, formation of AGS takes relatively much longer time while treating low-strength wastewaters like sewage. Strategies like increased volumetric flow by means of short cycles and mixing of sewage with industrial wastewaters can promote AGS formation while treating low-strength sewage. This article reviewed the state of research on AGS formation mechanisms, bioremediation capabilities and biotechnological applications of AGS technology in domestic and industrial wastewater treatment.

Biodegradation of tributyl phosphate, an organosphate triester, by aerobic granular biofilms.

Tributyl phosphate (TBP) is commercially used in large volumes for reprocessing of spent nuclear fuel. TBP is a very stable compound and persistent in natural environments and it is not removed in conventional wastewater treatment plants. In this study, cultivation of aerobic granular biofilms in a sequencing batch reactor was investigated for efficient biodegradation of TBP. Enrichment of TBP-degrading strains resulted in efficient degradation of TBP as sole carbon or along with acetate. Complete biodegradation of 2mM of TBP was achieved within 5h with a degradation rate of 0.4 µmol mL(-1) h(-1). TBP biodegradation was accompanied by release of inorganic phosphate in stoichiometric amounts. n-Butanol, hydrolysed product of TBP was rapidly biodegraded. But, dibutyl phosphate, a putative intermediate of TBP degradation was only partially degraded pointing to an alternative degradation pathway. Phosphatase activity was 22- and 7.5-fold higher in TBP-degrading biofilms as compared to bioflocs and acetate-fed aerobic granules. Community analysis by terminal restriction length polymorphism revealed presence of 30 different bacterial strains. Seven bacterial stains, including Sphingobium sp. a known TBP degrader were isolated. The results show that aerobic granular biofilms are promising for treatment of TBP-bearing wastes or ex situ bioremediation of TBP-contaminated sites.

Aerobic granular sludge mediated biodegradation of an organophosphorous ester, dibutyl phosphite.

Dibutyl phosphite, an organophosphorous compound, finds applications in different chemical industries and processes. Here, we report an efficient approach of biodegradation to be eventually used in bioremediation of dibutyl phosphite. Aerobic granules capable of dibutyl phosphite biodegradation were cultivated in a sequencing batch reactor (SBR). The SBR was operated with a 24-h cycle by feeding with dibutyl phosphite as a cosubstrate along with acetate. During the course of the SBR operation, aerobic granules of 0.9 ± 0.3 mm size were developed. Complete biodegradation of 1.4, 2 and 3 mM of dibutyl phosphite was achieved in 4, 5 and 8 h, respectively, accompanied by stoichiometric release of phosphite (H3 PO3). Phosphatase activity in the dibutyl phosphite-degrading granular biomass was 3- and 1.5-fold higher as compared to the activated sludge (seed biomass) and acetate-fed aerobic granules, respectively, indicating involvement in the hydrolysis of dibutyl phosphite. Microbial community analysis by t-RFLP showed the presence of 12 different bacterial types. Two bacterial strains capable of growth on dibutyl phosphite as sole carbon source were isolated and characterized as Acidovorax sp. and Sphingobium sp. The results show that aerobic microbial granules based process is suitable for the treatment of dibutyl phosphite contaminated water.

Efficient Computation of Small-Molecule Configurational Binding Entropy and Free Energy Changes by Ensemble Enumeration.

Here we present a novel, end-point method using the dead-end-elimination and A* algorithms to efficiently and accurately calculate the change in free energy, enthalpy, and configurational entropy of binding for ligand-receptor association reactions. We apply the new approach to the binding of a series of human immunodeficiency virus (HIV-1) protease inhibitors to examine the effect ensemble reranking has on relative accuracy as well as to evaluate the role of the absolute and relative ligand configurational entropy losses upon binding in affinity differences for structurally related inhibitors. Our results suggest that most thermodynamic parameters can be estimated using only a small fraction of the full configurational space, and we see significant improvement in relative accuracy when using an ensemble versus single-conformer approach to ligand ranking. We also find that using approximate metrics based on the single-conformation enthalpy differences between the global minimum energy configuration in the bound as well as unbound states also correlates well with experiment. Using a novel, additive entropy expansion based on conditional mutual information, we also analyze the source of ligand configurational entropy loss upon binding in terms of both uncoupled per degree of freedom losses as well as changes in coupling between inhibitor degrees of freedom. We estimate entropic free energy losses of approximately +24 kcal/mol, 12 kcal/mol of which stems from loss of translational and rotational entropy. Coupling effects contribute only a small fraction to the overall entropy change (1-2 kcal/mol) but suggest differences in how inhibitor dihedral angles couple to each other in the bound versus unbound states. The importance of accounting for flexibility in drug optimization and design is also discussed.

Design of mutation-resistant HIV protease inhibitors with the substrate envelope hypothesis.

There is a clinical need for HIV protease inhibitors that can evade resistance mutations. One possible approach to designing such inhibitors relies upon the crystallographic observation that the substrates of HIV protease occupy a rather constant region within the binding site. In particular, it has been hypothesized that inhibitors which lie within this region will tend to resist clinically relevant mutations. The present study offers the first prospective evaluation of this hypothesis, via computational design of inhibitors predicted to conform to the substrate envelope, followed by synthesis and evaluation against wild-type and mutant proteases, as well as structural studies of complexes of the designed inhibitors with HIV protease. The results support the utility of the substrate envelope hypothesis as a guide to the design of robust protease inhibitors.