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This study presents a novel approach to sustainable aquaculture by integrating biofloc technology (BFT) with a compact down-flow hanging sponge (DHS) reactor. The integrated BFT-DHS system effectively removed nitrogen compounds while maintaining ammonia-nitrogen (NH4+-N) concentrations below 1 mg-N L-1 without water exchange. Application of this system in a tank bred with juvenile Oreochromis niloticus showed a high NH4+-N removal rate of up to 97 % and nitrite (NO2- -N) concentrations were maintained at 0.1 ± 0.1 mg-N L-1. Microbial analysis revealed Gordonia as the predominant genus in the biofloc contributing to heterotrophic nitrification, while the Peptostreptococcaceae family dominated the DHS reactor. Heterotrophic nitrification seemed to be the primary process for enhanced nitrogen removal. Pathogenic bacteria, Vibrio sp. was absent throughout the study. This study highlights the potential integration of BFT and DHS system for sustainable aquaculture practice with effective nitrogen removal.

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Issues associated with accumulating microplastic (MP) in sewage sludge during wastewater treatment in a membrane bioreactor (MBR) system have not been studied in detail. Here, we investigated the microplastic's effects on floc characteristics, microbial community compositions, and fouling behavior inside sequencing-batch MBRs. MBRs were operated with 0, 7, 15, and 75 MPs/L of feed for 124-days. Results indicated that MP presence decreased sludge floc size, floc hydrophobicity, and extracellular polymeric substance (EPS) molecular size, and increased EPS concentration and the floc's negative zeta potential. These results were attributed to the facilitation of divalent cation (Ca2+ and Mg2+) uptake by MPs that weakened ion-bridging interactions within the sludge flocs. MPs accumulation slightly affected microbial structure and diversity. Relative abundances of dominant phyla, Actinobacteria, also decreased substantially. MPs also acted like a scouring material on membrane surfaces, inducing transformation of matured biofilm structures where protein content was substantially lower than nucleic acid content, in contrast to the control. Overall, MPs' negative effects on sludge flocs were counteracted by their scouring effect; consequently, SB-MBRs operated up to 4 months did not suffer from severe cake fouling, compared to control.

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Concrete cracks must be repaired promptly in order to prevent structural damage and to prolong the structural life of the building (or other such construction). Biological self-healing concrete is a recent alternative technology involving the biochemical reaction of microbial induced calcium carbonate precipitation (MICP). This study determined the most appropriate technique to encapsulate spores of Bacillus sphaericus LMG 22257 with sodium alginate so as to protect the bacterial spores during the concrete mixing and hardening period. Three techniques (extrusion, spray drying and freeze drying) to encapsulate the bacterial spores with sodium alginate were evaluated. The freeze-drying process provided the highest bacterial spore survival rate (100%), while the extruded and spray-dried processes had a lower spore survival rate of 93.8% and 79.9%, respectively. To investigate the viability of microencapsulated spores after being mixed with mortar, the decomposed urea analysis was conducted. The results revealed that the freeze-dried spores also showed the highest level of urea decomposition (metabolic activity assay used as a surrogate marker of spore germination and vegetative cell viability). Thus, the self-healing performance of concrete mixed with freeze-dried spores was evaluated. The results showed that the crack healing ratio observed from the mortar specimens with freeze-dried microencapsulated spores were significantly higher than those without bacteria. This study revealed that freeze drying has a high potential as a microencapsulation technique for application to self-healing concrete technology.

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A dissimilatory nitrate reduction to ammonium (DNRA) microbial community was developed under a high organic carbon to nitrate (C/NO3-) ratio in an anoxic semi-continuous sequencing batch reactor (SBR) fed with glucose as the source of carbon and NO3- as the electron acceptor. Activated sludge collected from a municipal wastewater treatment plant with good denitrification efficiency was used as the inoculum to start the system. The aim of this study was to examine the microbial populations in a high C/NO3- ecosystem for potential DNRA microorganisms, which are the microbial group with the ability to reduce NO3- to ammonium (NH4+). A low C/NO3- reactor was operated in parallel for direct comparisons of the microbial communities that developed under different C/NO3- values. The occurrence of DNRA in the high C/NO3- SBR was evidenced by stable isotope-labeled nitrate and nitrite (15NO3- and 15NO2-), which proved the formation of NH4+ from dissimilatory NO3-/NO2- reduction, in which both nitrogen oxides induced DNRA activity in a similar manner. An analysis of sludge samples with Illumina MiSeq 16S rRNA sequencing showed that the predominant microorganisms in the high C/NO3- SBR were related to Sulfurospirillum and the family Lachnospiraceae, which were barely present in the low C/NO3- system. A comparison of the populations and activities of the two reactors indicated that these major taxa play important roles as DNRA microorganisms under the high C/NO3- condition. Additionally, a beta-diversity analysis revealed distinct microbial compositions between the low and high C/NO3- SBRs, which reflected the activities observed in the two systems.

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A novel configured fluidized bed reactor (FBR) with granular rubber as the fluidized media was operated without internal recirculation to achieve denitrification. This FBR could operate under a low hydraulic retention time (HRT) of 50min due to the low rubber media density and absence of recirculation. Synthetic nitrate-rich wastewater with a fixed nitrate (NO3--N) concentration and varying COD concentrations was fed into the FBR. The nitrate removal profile showed a rapid nitrate reduction at the bottom of the reactor with a high performance under the low HRT. Different microbial communities were identified using Illumina Miseq sequencing. The dominant microorganisms belonged to the Beta- and Gamma-proteobacteria classes and played important roles in nitrate reduction. Acidovorax was abundant at low COD: NO3--N ratios, while Rhizobium and Zoogloea were dominant at high COD: NO3--N ratios. The COD: NO3--N ratio strongly influenced the composition of the microbial community including the dominant species.

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The effects of chitosan characteristics including the degree of deacetylation, molecular weight, particle size, pH pretreatment and immobilization time on the immobilization of nitrite-oxidizing bacteria (NOB) on biopolymeric chitosan were investigated. Nitrite removal efficiency of immobilized NOB depended on the degree of deacetylation, particle size, pH pretreatment on the surface of chitosan and immobilization time. Scanning electron microscope characterization illustrated that the number of NOB cells attached to the surface of chitosan increased with an increment of immobilization time. The optimal condition for NOB immobilization on chitosan was achieved during a 24-hr immobilization period using chitosan with the degree of deacetylation larger than 80% and various particle size ranges between 1-5 mm at pH 6.5. In general, the NOB immobilized on chitosan flakes has a high potential to remove excess nitrite from wastewater and aquaculture systems.

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A bottom substrate denitrification tank for a recirculating aquaculture system was developed. The laboratory scale denitrification tank was an 8 L tank (0.04 m2 tank surface area), packed to a depth of 5 cm with a bottom substrate for natural denitrifying bacteria. An aquarium pump was used for gentle water mixing in the tank; the dissolved oxygen in the water was maintained in aerobic conditions (e.g. > 2 mg/L) while anoxic conditions predominated only at the bottom substrate layer. The results showed that, among the four substrates tested (soil, sand, pumice stone and vermiculite), pumice was the most preferable material. Comparing carbon supplementation using methanol and molasses, methanol was chosen as the carbon source because it provided a higher denitrification rate than molasses. When methanol was applied at the optimal COD:N ratio of 5:1, a nitrate removal rate of 4591 +/- 133 mg-N/m2 tank bottom area/day was achieved. Finally, nitrate removal using an 80 L denitrification tank was evaluated with a 610 L recirculating tilapia culture system. Nitrate treatment was performed by batch transferring high nitrate water from the nitrification tank into the denitrification tank and mixing with methanol at a COD:N ratio of 5:1. The results from five batches of nitrate treatment revealed that nitrate was successfully removed from water without the accumulation of nitrite and ammonia. The average nitrate removal efficiency was 85.17% and the average denitrification rate of the denitrification tank was 6311 +/- 945 mg-N/m2 tank bottom area/day or 126 +/- 18 mg-N/L of pumice packing volume/day.

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A bacterial strain, WL(T), which was isolated from an activated sludge, was able to degrade alkylphenols. 16S rDNA sequence analysis indicated that strain WL(T) belonged to the genus Pseudomonas (sensu stricto) and formed a monophyletic clade with the type strain of Pseudomonas graminis and other members in the Pseudomonas putida subcluster with sequence similarity values higher than 97%. Genomic relatedness based on DNA-DNA hybridization of strain WL(T) to these strains is 2-41%. Strain WL(T) contained ubiquinone-9 as the main respiratory quinone, and the G+C content of DNA was 66 mol%. The organism contained hexadecanoic acid (16:0), hexadecenoic acid (16:1) and octadecenoic acid (18:1) as major cellular fatty acids. The hydroxy fatty acids detected were 3-hydroxydecanoic acid (3-OH 10:0), 3-hydroxydodecanoic acid (3-OH 12:0) and 2-hydroxydodecanoic acid (2-OH 12:0). These results, as well as physiological and biochemical characteristics clearly indicate that the strain WL(T) represents a new Pseudomonas species, for which the name Pseudomonas japonica is proposed. The type strain is strain WL(T) (=IAM 15071T=TISTR 1526T).

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