Physiological and biochemical response in green mussel Perna viridis subjected to continuous chlorination: Perspective on cooling water discharge criteria.

Heavy infestation by Perna viridis has been observed in the sub-seabed seawater intake tunnel and CWS of a tropical coastal power station in-spite of continuous low dose chlorination regime (0.2 ± 0.1 mg L-1) (CLDC), indicating periodical settlement and growth. Continuous arrival of mussels (colonized in the sub seabed tunnel intake section) at the pump house indicated that the mussels were able to tolerate and survive in a chlorinated environment, for varying time periods and were dislodged when they become weak and subsequent death, leading to flushing out of the system. In the present study, effect of continuous chlorination [0.2 mg L-1 (in-plant use); 0.5 mg L-1 (shock dose) & 1.0 mg L-1 (high levels)] was evaluated on mussels to assess; (a) time taken for mortality, (b) action of chlorine on physiological, genetic, metabolic and neuronal processes. 100% mortality of mussels was observed after 15 (0.2 mg L-1); 9 (0.5 mg L-1) and 6 days (1.0 mg L-1) respectively. Extended valve closure due to chlorination resulted in stress, impairing the respiratory and feeding behavior leading to deterioration in mussel health. Pseudofaeces excretion reduced to 68% (0.2 mg L-1); 10% (0.5 mg L-1) and 89% (1.0 mg L-1) compared to controls. Genotoxicity was observed with increase in % tail DNA fraction in all treatments such as 86% (0.2 mg L-1); 76% (0.5 mg L-1) and 85% (1.0 mg L-1). ROS stress biomarkers increased drastically/ peaked within the first 3 days of continuous chlorination with subsequent quenching by antioxidant enzymes. Gill produced highest generation of ROS; 38% (0.2 mg L-1); 97% (0.5 mg L-1); 98% (1.0 mg L-1). Additionally, it was shown that 84% (0.2 mg L-1), 72% (0.5 mg L-1), and 80.4% (1.0 mg L-1) of the neurotransmitter acetylcholinesterase activity was inhibited by chlorine at the nerve synapse. The cumulative impact of ROS generation, neuronal toxicity, and disrupted functions weakens the overall health of green mussels resulting in mortality.

Growth-dependent cr(VI) reduction by Alteromonas sp. ORB2 under haloalkaline conditions: toxicity, removal mechanism and effect of heavy metals.

Bacterial reduction of hexavalent chromium (VI) to chromium (III) is a sustainable bioremediation approach. However, the Cr(VI) containing wastewaters are often characterized with complex conditions such as high salt, alkaline pH and heavy metals which severely impact the growth and Cr(VI) reduction potential of microorganisms. This study investigated Cr(VI) reduction under complex haloalkaline conditions by an Alteromonas sp. ORB2 isolated from aerobic granular sludge cultivated from the seawater-microbiome. Optimum growth of Alteromonas sp. ORB2 was observed under haloalkaline conditions at 3.5-9.5% NaCl and pH 7-11. The bacterial growth in normal culture conditions (3.5% NaCl; pH 7.6) was not inhibited by 100 mg/l Cr(VI)/ As(V)/ Pb(II), 50 mg/l Cu(II) or 5 mg/l Cd(II). Near complete reduction of 100 mg/l Cr(VI) was achieved within 24 h at 3.5-7.5% NaCl and pH 8-11. Cr(VI) reduction by Alteromonas sp. ORB2 was not inhibited by 100 mg/L As(V), 100 mg/L Pb(II), 50 mg/L Cu(II) or 5 mg/L Cd(II). The bacterial cells grew in the medium with 100 mg/l Cr(VI) contained lower esterase activity and higher reactive oxygen species levels indicating toxicity and oxidative stress. In-spite of toxicity, the cells grew and reduced 100 mg/l Cr(VI) completely within 24 h. Cr(VI) removal from the medium was driven by bacterial reduction to Cr(III) which remained in the complex medium. Cr(VI) reduction was strongly linked to aerobic growth of Alteromonas sp. The Cr(VI) reductase activity of cytosolic protein fraction was pronounced by supplementing with NADPH in vitro assays. This study demonstrated a growth-dependent aerobic Cr(VI) reduction by Alteromonas sp. ORB2 under complex haloalkaline conditions akin to wastewaters.

Pilot-scale aerobic granular sludge reactors with granular activated carbon for effective nitrogen and phosphorus removal from domestic wastewater.

Aerobic granular sludge (AGS) is a breakthrough biotechnology of 21st century and an innovative alternative to activated sludge for treating wastewater. Concerns on long-start up periods for development of AGS and stability of granules are impeding its widespread implementation for treating low-strength domestic wastewater especially in tropical climate conditions. Addition of nucleating agents have been shown to improve development of AGS while treating low-strength wastewaters. There are no previous studies on AGS development and biological nutrient removal (BNR) in the presence of nucleating agents during treatment of real domestic wastewater. This study investigated AGS formation and BNR pathways while treating real domestic wastewater in a 2 m3 pilot-scale granular sequencing batch reactor (gSBR) operated without and with granular activated carbon (GAC) particles. The gSBRs were operated under tropical climate (T ≈ 30 °C) for >4-years to evaluate the effect of GAC addition on granulation, granular stability and BNR at pilot-scale. Formation of granules was observed within 3 months. MLSS values of 4 and 8 g/L were recorded within 6 months in gSBRs without and with GAC particles, respectively. The granules had an average size of 1.2 mm and SVI5 of 22 mL/g. Ammonium was mainly removed through nitrate formation in the gSBR without GAC. But, ammonium was removed by short-cut nitrification via nitrite due to washout of nitrite oxidizing bacteria in the presence of GAC. Phosphorus removal was much higher in gSBR with GAC due to the establishment of enhanced biological phosphorus removal (EBPR) pathway. After 3 months, the phosphorus removal efficiencies were at 15 % and 75 %, respectively, without and with GAC particles. The addition of GAC led to moderation in bacterial community and enrichment of polyphosphate-accumulating organisms. This is the first ever report on pilot-scale demonstration of AGS technology in the Indian sub-continent and GAC addition on BNR pathways.

Selenite reduction and biogenesis of selenium-nanoparticles by different size groups of aerobic granular sludge under aerobic conditions.

Microbial transformations play a vital role in Se cycle in the environment and decrease the solubility and toxicity of Se oxyanions by converting to elemental selenium (Se0) nanostructures. Aerobic granular sludge (AGS) has attracted interest due to efficient reduction of selenite to biogenic Se0 (Bio-Se0) and retention in bioreactors. Here, selenite removal, biogenesis of Bio-Se0 and entrapment of Bio-Se0 by different size groups of aerobic granules were investigated to optimize biological treatment process for Se-laden wastewaters. Furthermore, a bacterial strain showing high selenite tolerance and reduction was isolated and characterized. Removal of selenite and conversion to Bio-Se0 were achieved by all the size groups of granules ranging from 0.12 mm to 2 mm and above. However, selenite reduction and Bio-Se0 formation were rapid and more efficient with large aerobic granules (≥0.5 mm). The formed Bio-Se0 was majorly associated with the large granules, due to better entrapment capabilities. In contrast, the Bio-Se0 formed by the small granules (≤0.2 mm) was distributed both in the granules and aqueous phase because of ineffective entrapment. Scanning electron microscope and energy dispersive X-ray (SEM-EDX) analysis confirmed formation of Se0 spheres and association with the granules. Efficient selenite reduction and entrapment of Bio-Se0 was related to prevalent anoxic/anaerobic zones in the large granules. A bacterial strain showing efficient SeO32- reduction of up to 15 mM SeO32- under aerobic conditions was identified as Microbacterium azadirachtae. SEM-EDX analysis confirmed the formation and entrapment of Se0 nanospheres (size: 100 ± 5 nm) in the extracellular matrix. The cells immobilized in alginate beads showed effective SeO32- reduction and Bio-Se0 entrapment. Efficient reduction and immobilization of bio-transformed metalloids by large AGS and AGS-borne bacteria implicates prospective use in bioremediation of metal(loid) oxyanions and bio-recovery.

De novo granulation of sewage-borne microorganisms: A proof of concept on cultivating aerobic granular sludge without activated sludge and effective enhanced biological phosphorus removal.

Long start-up periods for granulating activated sludge and concerns on granular stability are the bottlenecks reported during implementation of novel aerobic granular sludge (AGS) technology in municipal wastewater treatment plants. Here, de novo granulation of sewage-borne microorganisms without using activated sludge (AS) inoculum was investigated in bench-scale sequencing batch reactors (SBR). Data showed that formation of AGS from sewage-borne microorganisms was rapid and first granules appeared within one week. Granulation was indicated by appearance of biomass particles (size >0.12 mm), high biomass levels (∼8 g/L) and superior settling properties (SVI30 min: 30 mL/g). Granulation process involved distinct stages like formation of aggregates, retention of aggregates, and growth of millimetre sized granules. Simultaneous COD, nitrogen and phosphorous removal was established within 10 days of start-up in the SBR without using AS inoculum. However, phosphorus removal became stable after 50 days of start-up. Total nitrogen (TN) and total phosphorus (TP) removals of 92% and 70%, respectively, were achieved from real domestic wastewater. Furthermore, addition of granular activated carbon (GAC) had improved both granulation and biological nutrient removals. Interestingly, phosphorus removal became quite stable within 10 days of start-up in the SBR operated with GAC particles. TN and TP removals were found to be higher at >98% and >94%, respectively, in GAC-augmented SBR. Removal of ammonia and phosphorus were mediated by nitritation-denitritation and enhanced biological phosphorus removal (EBPR) pathways, respectively. The bacterial diversity of AGS was lower than that of sewage. Quantitative PCR indicated enrichment of ammonia oxidizing bacteria, denitrifying bacteria and polyphosphate accumulating organisms during granulation. De novo granulation of sewage-borne microorganisms is a promising approach for rapidly cultivating AGS and establishing biological nutrient removal in sewage treatment plants.

Fungal infections: Pathogenesis, antifungals and alternate treatment approaches.

Increasing incidence of fungal infections of recent times requires immediate intervention. Fungal infections are seldom construed at initial stages that intensify the severity of infections and complicate the treatment procedures. Fungal pathogens employ various mechanisms to evade the host immune system and to progress the severity of infections. For the treatment of diverse superficial and systemic infections, antifungal drugs from the available repertoire are administered. However, well documented evidence of fungal resistance to most of the antifungal drugs hampers disease control and poses challenges in antifungal therapy. Several physiological adaptations and genetic mutations followed by their selection in presence of antifungal agents drive the resistance development in fungi. The availability of limited antifungal arsenal, emergence of resistance and biofilm-conferred resistance drives the need for development of novel drugs and alternate approaches for the better treatment outcome against mycoses. This graphical review explicitly shed light on various fungal infections and causative organisms, pathogenesis, different antifungal drugs and resistance mechanisms including host immune response and evasion strategies. Here, we have highlighted recent developments on novel antifungal agents and other alternate approaches for fighting against fungal infections.

Effects of oxytetracycline on aerobic granular sludge process: Granulation, biological nutrient removal and microbial community structure.

Formation of aerobic granular sludge (AGS), process performance and microbial community structure were investigated in lab-scale sequencing batch reactors (SBR) operated without and with oxytetracycline (OTC). Granulation of activated sludge and appearance of AGS was observed in parallel SBRs operated without and with OTC. However, formation of well-settling aerobic granules was relatively faster in the SBR fed with 100 µg/L OTC and observed within 2 weeks of start-up. Ammonium, total nitrogen, and phosphorus removals were quickly established in the AGS cultivated without OTC. In contrast, nitrogen and phosphorus removals were lower in the OTC fed SBR. But, a gradual improvement in nitrogen and phosphorus removals was observed. After 45 days, nitrogen and phosphorous removals were stabilized at 99% and 70%, respectively, due to establishment of OTC-tolerant community. qPCR revealed the impact of OTC on ammonium oxidizing bacteria, polyphosphate accumulating organisms and their enrichment during exposure to OTC. Ammonium and phosphorus were majorly removed via nitritation-denitritation and enhanced biological phosphorus removal (EBPR) pathways, respectively, in the presence of OTC. Brevundimonas (35%), Thaurea (14%) sp. Ca. Competibacter (5.6%), and Ca. Accumulibacter (4.2%) were enriched in OTC-fed AGS. Of the two OTC-tolerant strains isolated, Micrococcus luteus exhibited growth and efficient OTC biotransformation at different OTC concentrations. Moreover, M. luteus was predominantly growing in the form of aggregates. Key traits such as tolerance, biotransformation and high autoaggregation ability allowed a niche for this strain in the granules. This work has important implications in understanding the effect of antibiotics on AGS and designing AGS based treatment for antibiotic-laden wastewaters.

Enhanced biological phosphorus removal in aerobic granular sludge reactors by granular activated carbon dosing.

This study investigated the effects of granular activated carbon (GAC) addition on the enrichment of polyphosphate accumulating organisms (PAOs), stratification of PAOs in the co-existing GAC-biofilms and granules and biological nutrient removal (BNR) in aerobic granular sludge (AGS) reactors. It was found that BNR increased in the GAC-augmented system. Establishment of enhanced biological phosphorus removal (EBPR) pathway was faster with about 1.7 to 2-fold higher P removal in GAC system than control. EBPR biomass grown in the presence of GAC was segregated into different size fractions for determining BNR and stratification of microbial groups. It was found that EBPR was majorly associated with the large biomass (>0.5 mm) fraction, corroborating with higher abundance of PAOs. Higher P removals of 60 to 70% with characteristic EBPR profiles were observed in 0.5 mm fraction. In contrast, P removals by 0.25 mm fraction were lower at 20 to 35% without EBPR profiles. EBPR biomass (>0.5 mm) fraction was segregated into granules and GAC-biofilms for determining the role of GAC in PAOs enrichment. P release (2.5-3.5 mg L-1 P) and P uptake (5-7 mg L-1 P) were higher in the P removal profiles exhibited by GAC-biofilms. In contrast, P release and P uptake were lower with the granules. These differences in P removal profiles resulted in distinct net P removal efficiencies of 70 ± 5% and 50 ± 6% for GAC-biofilms and granules, respectively. These differences in P removals were corroborated by higher abundance of PAOs in the GAC-biofilms than co-existing granules. PAO clade-level enrichment was found to be dependent on substrate wherein acetate feeding enriched PAO clade I, while acetate-propionate feeding caused enrichment of both PAO clade I and II. These results suggest that GAC addition to AGS reactors can aid in enrichment of PAOs, reduce the start-up period for EBPR, and increase P removal efficiencies.

Aerobic granular sludge for efficient biotransformation of chalcogen SeIV and TeIV oxyanions: Biological nutrient removal and biogenesis of Se0 and Te0 nanostructures.

Simultaneous removal of selenite (SeIV), tellurite (TeIV) and nutrients by aerobic granular sludge (AGS) was investigated. A sequencing batch reactor (SBR) was operated with increasing SeIV and TeIV (up to 500 µM each) for 205 days to evaluate metalloid oxyanion and nutrient removal. AGS efficiently removed SeIV and TeIV by readily converting them to biomass associated forms. The total Se and Te removal efficiencies were higher at 98% and 99%, respectively. Formation of biomass-associated Se0 and Te0 was confirmed by XRD, Raman spectroscopy and SEM-EDX. Feeding of SeIV and TeIV elicited inhibitory action on ammonium removal initially, nonetheless removal performance was recovered during the subsequent cycles. Ammonium, total nitrogen and phosphorus removals were stabilized at 85%, 80% and 75%, respectively, at 500 µM of SeIV and TeIV. Sequencing of 16S rRNA gene confirmed enrichment of known SeIV and TeIV reducing bacteria in the granules. qPCR and removal kinetics supported ammonia removal via nitritation-denitritation. This work demonstrates functional capabilities of AGS for effectively removing toxic SeIV and TeIV oxyanions apart from performing simultaneous COD, nitrogen and phosphorus removal. Efficient biological nutrient removal in the presence of toxic SeIV and TeIV concentrations, suggests robustness of AGS and its resilience to toxic contaminants.

Assessment of alkylimidazolium chloride ionic liquid formulations for cleaning and disinfection of environmental surfaces.

BACKGROUND: Surface disinfection is fundamental to good environmental hygiene and preventing infections. Development of newer formulations that can effectively kill and remove microorganisms from the surfaces is desired. METHODS: Here, we assessed the efficacy of 1-hexadecyl-3-methylimidazolium chloride [C16MIM][Cl] ionic liquid (IL) and its formulation in ethanol for killing and removing bacteria from different environmental surfaces. Efficacy of IL and its formulation was determined on known monospecies bacterial cultures and unknown multispecies bacterial cultures on environmental surfaces. RESULT: The surface disinfection efficacy of [C16MIM][Cl] was concentration dependent and achieved 41 to 100% reduction in total viable bacterial counts of Gram positive and Gram negative bacteria at varied concentrations. The treatment of wooden surface with 0.1% [C16MIM][Cl] caused 98% reduction in bacterial load within 20 s contact time as against mere 45% reduction (20 s) with 70% ethanol. Antibacterial and surface disinfection activities of [C16MIM][Cl] have increased markedly when prepared in 70% ethanol, suggesting synergistic activity. A formulation comprising of 0.01% [C16MIM][Cl] in 70% ethanol showed effective surface disinfection and achieved 95% to 98% reduction in bacterial load on different surfaces. CONCLUSION: Ionic liquids are potent candidates for disinfection of environmental surfaces.

Enhancing biological nitrogen and phosphorus removal performance in aerobic granular sludge sequencing batch reactors by activated carbon particles.

Long start-up periods for aerobic granular sludge (AGS) formation and establishment of P removal pathways are challenges for widespread implementation of AGS process. External additives such as activated carbon (AC) attracted interest for accelerating AGS formation. However, the roles of AC in granulation and biological nutrient removal (BNR) are not understood. Here, the role of AC was investigated in decreasing start-up periods in AGS formation and BNR under different carbon substrate conditions (i.e., acetate (HAc), propionate (HPr) and HAc-HPr) in sequencing batch reactors (SBRs). AC addition increased aggregation index and settleability of activated sludge (AS) inoculum which minimized AS washout from SBRs. AC addition hastened AGS formation and establishment of BNR pathways by facilitating AS retention and biofilm formation. Feeding HAc or HAc-HPr supported better granulation (MLSS: 6-7 g l-1, SVI: 30-40 ml g-1) than HPr (MLSS: 4 g l-1, SVI: 70). The start-up periods for efficient total nitrogen (TN) removals were decreased to 22 and 16 d from 38 to 25 d, respectively, in AC augmented SBRs fed with either HAc or HAc-HPr. TN removals were higher at ≥95% in HAc or HAc-HPr fed SBRs. Total phosphorus (TP) removals were also higher in AC-augmented SBRs at 80% and ≥90% in HAc and HAc-HPr fed SBRs, respectively. In contrast, TN and TP removals were lower at 70% and 35%, respectively, in HPr fed SBR. Ammonium was primarily removed via nitritation-denitritation pathway. Phosphorus removal was at 1.7 to 2-fold higher in AC augmented SBRs and driven by enhanced biological phosphorus removal (EBPR) pathway. MiSeq sequencing and qPCR revealed higher enrichment of polyphosphate accumulating organisms (PAOs), denitrifying PAOs, and ammonia oxidizers in AC-augmented SBRs fed with HAc or HAc-HPr. This study demonstrates that AC addition can be considered for enrichment of PAOs and establishment of EBPR in aerobic granular SBRs.

Development of biogenic bimetallic Pd/Fe nanoparticle-impregnated aerobic microbial granules with potential for dye removal.

The objective of this study was to develop bimetallic core-shell Pd/Fe nanoparticles on the surface of aerobic microbial granules (Bio-Pd/Fe) and to evaluate their dye removal potential using a representative dye, methyl orange (MO). The aerobic microbial granules (1.5 ± 0.32 mm) were grown for 70 days in a 3-L glass sequencing batch reactor (SBR) with a 12-h cycle time. The Bio-Pd/Fe formation was catalyzed by the Bio-H2 gas produced by the granules. The developed Bio-Pd/Fe was further used for MO removal from aqueous solutions, and the reaction parameters were optimized by response surface methodology (RSM). The XRD, SEM, EDAX, elemental mapping, and XPS studies confirmed the formation of Bio-Pd/Fe. Under the optimized removal conditions, 99.33% MO could be removed by Bio-Pd/Fe, whereas removal by Bio-Pd, Bio-Fe, aerobic microbial granules, and heat-killed granules were found to be quite low (68.91 ± 0.2%, 76.8 ± 0.3%, 19.8 ± 0.6%, and 6.59 ± 0.2%, respectively). The mechanism of removal was investigated by UV-visible spectroscopy, redox potential analysis, HR-LCMS analyses of the solution phase, and XRD and XPS analyses of the solid sorbent. The degradation products of MO exhibited m/z values corresponding to 292, 212, and 160 m/z. The remnant toxicity of the intermediate degradation products was analysed using freshwater algae, Scenedesmus sp. And Allium cepa, as indicator organisms. These assays suggested that after the treatment with Bio-Pd/Fe, MO was transformed to a lesser toxic form.

Cathodic selenium recovery in bioelectrochemical system: Regulatory influence on anodic electrogenic activity.

Metal(loid)s are used in various industrial activities and widely spread across the environmental settings in various forms and concentrations. Extended releases of metal(loid)s above the regulatory levels cause environmental and health hazards disturbing the ecological balance. Innovative processes for treating the metal(loid)-contaminated sites and recovery of metal(loid)s from disposed waste streams employing biotechnological routes provide a sustainable way forward. Conventional metal recovery technologies demand high energy and/or resource inputs, which are either uneconomic or unsustainable. Microbial electrochemical systems are promising for removal and recovery of metal(loid)s from metal(loid)-laden wastewaters. In this communication, a bioelectrochemical system (BES) was designed and operated with selenium (Se) oxyanion at varied concentrations as terminal electron acceptor (TEA) for reduction of selenite (Se4+) to elemental selenium (Se0) in the abiotic cathode chamber. The influence of varied concentrations of Se4+ towards Se0 recovery at the cathode was also evaluated for its regulatory role on the electrometabolism of anode-respiring bacteria. This study observed 26.4% Se0 recovery (cathode; selenite removal efficiency: 73.6%) along with organic substrate degradation of 74% (anode). With increase in the initial selenite concentration, there was a proportional increase in the dehydrogenase activity. Bioelectrochemical characterization depicted increased anodic electrogenic performance with the influence of varied Se4+ concentrations as TEA and resulted in a maximum power density of 0.034 W/m2. The selenite reduction (cathode) was evaluated through spectroscopic, compositional and structural analysis. X-ray diffraction and Raman spectroscopy showed the amorphous nature, while Energy Dispersive X-ray spectroscopy confirmed precipitates of the deposited Se0 recovered from the cathode chamber. Scanning electron microscopic images clearly depicted the Se0 depositions (spherical shaped; sized approximately 200 nm in diameter) on the electrode and cathode chamber. This study showed the potential of BES in converting soluble Se4+ to insoluble Se0 at the abiotic cathode for metal recovery.

Biological nutrient removal by halophilic aerobic granular sludge under hypersaline seawater conditions.

Biological nutrient removal and physical properties of halophilic aerobic granular sludge (hAGS) cultivated from autochthonous seawater-born microbes were investigated under hypersaline seawater conditions. hAGS achieved stable total nitrogen (TN) and total phosphorus (TP) removals of 96 ± 3% and 95 ± 4%, respectively, from seawater-based wastewater at 3.4% salt. At 4 to 12% salt concentrations, stable TN and TP removals of 82-99% and 95-96%, respectively, were maintained over 4 months under seawater conditions. Ammonium and phosphorus were mainly removed by nitritation-denitritation and enhanced biological phosphorus removal pathways, respectively. Stappiaceae (45%) and Rhodobacteraceae (21%) were the dominant genera in hAGS performing nutrient removal at 12% salt. hAGS contained acid-soluble extracellular polymeric substance as the major structural polymer which increased from 0.43 ± 0.02 g/gTS at 3.4% salt to 0.93 ± 0.03 g/gTS at 12% salt. Cultivation of hAGS from autochthonous wastewater-microbes can be a promising approach for achieving biological nitrogen and phosphorus removals from hypersaline seawater-based wastewaters.

Alkylimidazolium Ionic Liquids as Antifungal Alternatives: Antibiofilm Activity Against Candida albicans and Underlying Mechanism of Action.

Candida albicans is an opportunistic pathogen causes fungal infections that range from common skin infections to persistent infections through biofilm formation on tissues, implants and life threatening systemic infections. New antifungal agents or therapeutic methods are desired due to high incidence of infections and emergence of drug-resistant strains. The present study aimed to evaluate (i) the antifungal and antibiofilm activity of 1-alklyl-3-methyl imidazolium ionic liquids ([CnMIM]+[X]-, n = 4, 12 and 16) against Candida albicans ATCC 10231 and two clinical C. albicans strains and (ii) the mechanism of action of promising antifungal ionic liquid on C. albicans. Two of the tested compounds were identified as more effective in preventing growth and biofilm formation. These ionic liquid compounds with -dodecyl and -hexadecyl alkyl groups effectively prevented biofilm formation by fluconazole resistant C. albicans 10231 and two other clinical C. albicans strains. Although both the compounds caused viability loss in mature C. albicans biofilms, an ionic liquid with -hexadecyl group ([C16MIM]+[Cl]-) was more effective in dispersing mature biofilms. This promising ionic liquid compound ([C16MIM]+[Cl]-) was chosen for determining the underlying mode of action on C. albicans cells. Light microscopy showed that ionic liquid treatment led to a significant reduction in cell volume and length. Increased cell membrane permeability in the ionic liquid treated C. albicans cells was evident in propidium iodide staining. Leakage of intracellular material was evident in terms of increased absorbance of supernatant and release of potassium and calcium ions into extracellular medium. A decrease in ergosterol content was evident when C. albicans cells were cultured in the presence of antifungal ionic liquid. 2',7'-Dichlorodihydrofluorescein acetate assay revealed reactive oxygen species generation and accumulation in C. albicans cells upon treatment with antifungal ionic liquid. The effect of antifungal ionic liquid on mitochondria was evident by decreased membrane potential (measured by Rhodamine 123 assay) and loss of metabolic activity (measured by MTT assay). This study demonstrated that imidazolium ionic liquid compound exert antifungal and antibiofilm activity by affecting various cellular processes. Thus, imidazolium ionic liquids represent a promising antifungal treatment strategy in lieu of resistance development to common antifungal drugs.

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.

Textile dye biodecolourization and ammonium removal over nitrite in aerobic granular sludge sequencing batch reactors.

Biodecolourization of azo dye and removal of ammonium by aerobic granular sludge (AGS) was investigated under different growth conditions. AGS not previously exposed to azo dye was able to effectively decolourize azo dye under anaerobic and microaerophilic conditions. Azo dye, total organic carbon and ammoniacal nitrogen removal efficiencies of 89-100%, 79-95% and 92-100%, respectively, were achieved in the AGS reactor operated for 80days under microaerophilic conditions. Removal of carbon, nitrogen and phosphorus was not impacted by azo dye loading. Azo dye, organic carbon and ammonium were majorly removed in the anoxic period wherein bulk dissolved oxygen was ranged from 0.5 and <0.08mgL-1. Removal of 60mgL-1 NH4+-N was associated only with smaller amounts of nitrite build-up (∼5mgL-1 NO2--N) and negligible nitrate concentrations. Profiles of nitrogen compounds in individual sequencing batch reactor cycles supported the occurrence of ammonium removal over nitrite pathway. Bacterial community analysis showed enrichment of specific microorganisms capable of decolourizing azo dyes in the dye-decolourizing AGS. Dye decolourization and nutrient removal by AGS under microaerophilic conditions is a novel finding and can be further developed for treating textile wastewaters onsite or after dilution with sewage.

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.

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.

Selenite reduction and ammoniacal nitrogen removal in an aerobic granular sludge sequencing batch reactor.

Simultaneous removal of selenite and ammonium by aerobic granular sludge was investigated to develop an improved biological treatment process for selenium rich wastewaters. Aerobic granules not previously exposed to selenite were able to remove selenite by converting it to elemental selenium (Se(0)) and simultaneously remove ammonium under different conditions in batch experiments. To achieve sustainable selenite and ammonium removal, an aerobic granular sludge reactor was operated in fill-and-draw mode with a cycle of anaerobic (8 h) and aeration (15 h) phases. Almost complete removal of different initial concentrations of selenite up to 100 µM was achieved in the anaerobic phase. Ammonium removal was severely inhibited when the granules were initially exposed to 1.27 mg L-1 selenite, but ammonium and total nitrogen removal efficiencies gradually improved to 100 and 98%, respectively, under selenite-reducing conditions. Selenite loading shifted ammonium removal occurring mainly during the anaerobic phase to both the anaerobic and aeration phases. Selenite was removed from the aqueous phase by converting it to nanoparticulate Se(0), which was entrapped in the granular sludge. Scanning electron microscop-energy dispersive X-ray spectroscopy and X-ray diffraction analysis confirmed the formation of Se(0) nanospheres and their retention in the granular sludge. The effluent Se ranged from 0.02 to 0.25 mg Se L-1, while treating up to 12.7 mg L-1 selenite, which is lower as compared to previous studies on selenite removal using activated sludge or anaerobic granular sludge. This study shows that aerobic granular sludge reactors are not only capable of removing toxic selenite, but offer improved treatment of Se-rich wastewaters.