Multilayer immobilizing of denitrifying Bacillus sp. and TiO2-AgNPs on floating expanded clay carrier for co-treatment of nitrite and pathogens in aquaculture.

Nitrite contamination and the spread of pathogens can seriously degrade water quality. To simultaneously control these factors, an innovative approach of fabricating a remediation agent that contained denitrifying bacteria and TiO2-AgNPs co-immobilized on floating expanded clay (EC) was proposed in this study. The EC was fabricated from a mixture of clay and rice husk through pyrolysis at a high temperature of 1200 °C, followed by a rapid cooling step to create a porous structure for the material. TiO2NPs were modified with Ag to shift the absorbance threshold of TiO2-AgNPs into the visible region of 700-800 nm. The experimental results showed that the stirring speed of 250 rpm was suitable for immobilizing TiO2-AgNPs on EC and achieved the highest Ti and Ag content of 639.38 ± 3.04 and 200.51 ± 3.71 ppm, respectively. Coating TiO2-Ag/EC with chitosan (0.5%) significantly reduced the detachment level of immobilized TiO2-AgNPs compared to that of the material with no coating. In particular, this functionalized material inhibited 99.93 ± 0.1% of Vibrio parahaemolyticus pathogen but did not adversely affect the denitrifying bacteria after 2 h of visible light irradiation. Based on the electrostatic bond between oppositely charged polymers, the denitrifying bacteria, Bacillus sp., in alginate solution was successfully immobilized on the chitosan-coated TiO2-Ag/EC with a bacteria density of (76.67 ± 9.43) × 107 CFU g-1, retaining its nitrite removal efficiency at 99.0 ± 0.27% through six treatment cycles. These findings provide solid evidence for further investigating the combination of biodegradation and photodegradation in wastewater treatment.

Allium sativum@AgNPs and Phyllanthus urinaria@AgNPs: a comparative analysis for antibacterial application.

Although medicinal herbs contain many biologically active ingredients that can act as antibiotic agents, most of them are difficult to dissolve in lipids and absorb through biofilms in the gastrointestinal tract. Besides, silver nanoparticles (AgNPs) have been widely used as a potential antibacterial agent, however, to achieve a bactericidal effect, high concentrations are required. In this work, AgNPs were combined into plant-based antibiotic nanoemulsions using biocompatible alginate/carboxyl methylcellulose scaffolds. The silver nanoparticles were prepared by a green method with an aqueous extract of Allium sativum or Phyllanthus urinaria extract. The botanical antibiotic components in the alcoholic extract of these plants were encapsulated with emulsifier poloxamer 407 to reduce the particle size, and make the active ingredients both water-soluble and lipid-soluble. Field emission scanning electron microscopy (FESEM) and energy-dispersive X-ray (EDX) analysis showed that the prepared nanosystems were spherical with a size of about 20 nm. Fourier transform infrared spectroscopy (FTIR) confirmed the interaction of the extracts and the alginate/carboxyl methylcellulose carrier. In vitro drug release kinetics of allicin and phyllanthin from the nanosystems exhibited a retarded release under different biological pH conditions. The antimicrobial activity of the synthesized nanoformulations were tested against Escherichia coli. The results showed that the nanosystem based on Allium sativum possesses a significantly higher antimicrobial activity against the tested organisms. Therefore, the combination of AgNPs with active compounds from Allium sativum extract is a good candidate for in vivo infection treatment application.

Characterization of Isolated Aerobic Denitrifying Bacteria and Their Potential Use in the Treatment of Nitrogen-Polluted Aquaculture Water.

Ammonia and nitrite treatments are the critical steps that must be done to ensure the healthy growth of aquatic animals. The nitrification process is often used for nitrogen removal purposes due to its efficiency and environmentally friendly properties. However, the varied growth rate between ammonia and nitrite-oxidizing bacteria can cause nitrite accumulation, leading to the massive mortality of aquatic animals at high concentrations. Therefore, this study aimed to integrate the fast-growing heterotrophic nitrite-reducing bacteria with nitrifying bacteria to achieve a quicker nitrite removal activity. The two denitrifying Bacillus sp. ST20 and Bacillus sp. ST26 were screened from shrimp ponds in Bac Lieu province, Vietnam. Obtained results showed that under anoxic conditions, the nitrite removal efficiency of these two strains reached 68.5-82% at nitrite initial concentration of 20 mgN-NO2/L after 72 h. Higher efficiency of over 95% was gained under oxic conditions. Hence, it enabled the use of denitrifying and nitrifying bacteria-co-immobilized carriers for ammonia oxidation and nitrite reduction simultaneously in a single-aerated bioreactor. A total of over 96% nitrogen content was removed during the bioreactor operation, despite the increase of inputting nitrogen concentration from 40 to 200 mgN/L. Moreover, the suitable conditions for bacterial growth and nitrite conversion activity of the ST20 and ST26 were detected as 15‰ salinity and 35 °C. The isolates also utilized various C-sources for growth, hence widening their applicability. The present study suggested that the isolated aerobic denitrifying bacteria are potentially used for the complete removal of nitrogen compounds from aquaculture wastewater.

Modification of expanded clay carrier for enhancing the immobilization and nitrogen removal capacity of nitrifying and denitrifying bacteria in the aquaculture system.

In aquaculture systems, the treatment of nitrogen pollution has always been a center of attention due to its impact on productiveness. The bioremediation method based on simultaneous nitrification and denitrification was often used to effectively remove ammonium, nitrite, and nitrate compounds. In addition, the attachment and biofilm formation of the nitrogen-converting bacteria on carriers had superior removal efficiency over the suspended bacteria. Thus, this study focused on the fabrication of a porosity floatable expanded clay (EC) carrier that provided the basic structure for the immobilization of the nitrifiers Nitrosomonas sp., Nitrobacter sp., and the denitrifier Bacillus sp. The EC was also coated with alginate and essential nutrient to support the cohesion and growth of bacteria. Especially, the selected Bacillus sp. previously proved was able to reduce nitrite/nitrate in aerobic conditions. The co-immobilization of these three aerobic bacteria on the prepared carrier would simply the treatment process in practical use. Initial results showed that the integration of essential nutrients (N, P, K) on alginate coated EC (EC_Alg_N) increased bacterial density to (57 ± 3) × 107 - (430 ± 30) × 108 CFU/g, which then led to the enhancement of removal efficiency up to 91.62 ± 0.67% in the medium containing initial nitrogen content of 60 mg-N/L. The nitrogen removal efficacy of bacterial immobilized EC_Alg_N remained at 83.95 ± 0.15% after being reused for 6 cycles. In conclusion, the bacterial immobilized EC_Alg_N could be a potential material for nitrogen polluted wastewater treatment in aquaculture systems.