Search Results (112)

The recovery of extracellular polymeric substances (EPS) from activated sludge (AS) represents a promising strategy to transform wastewater treatment plants (WWTPs) into resource recovery facilities within a circular economy framework. In this study, EPS was extracted from an AS process in a full-scale WWTP, highlighting its catalytic and bioflocculant properties, which represent an innovation in the valorization of this biopolymer. The EPS was subsequently characterized in terms of polysaccharides, proteins, and enzymatic activities (amylase and lipase). The bioflocculation performance of the EPS was evaluated using activated sludge mixed liquor. Results showed that EPS recovery yields using 50 °C and 80 °C were 196.3 ± 38.2 mg EPS/g sludge and 283.5 ± 85.4 mg EPS/g sludge, respectively. Enzymatic assays confirmed amylase activity ranging from 100 to 350 U/g sludge according to the extraction temperature. Lipolytic activity (20 U/g sludge) was comparable to values reported in the literature for EPS from biological sludge. The addition of EPS significantly improved the sludge settling velocity (from 0.86 to 4.48 m/h) and the sludge volume index (from 118.6 to 35.5). However, EPS application also increased the resistance to filtration by 50% and reduced cellular respiration by approximately 40%. Overall, the findings demonstrate that EPS from activated sludge acts as an effective bioflocculant with relevant catalytic properties, highlighting its potential as a high-value biotechnological product while also pointing to operational challenges that require further optimization. Full article

Evaluation of characteristics and flocculation mechanisms of microbial flocculants facilitates the identification of potential applications and informs the fine-tuning of operational conditions for maximum activity. Therefore, this study aimed to characterise and optimise the operational conditions of bioflocculants produced from Klebsiella pneumoniae and Meyerozyma guilliermondii for potent wastewater treatment. Scanning electron microscopy, X-ray Diffraction (XRD), Thermogravimetric Analysis (TGA) and Fourier Transform Infrared Spectroscopy (FTIR) were employed to assess the surface morphology, crystalline structure, thermal stability, and functional group composition of the bioflocculants. Their cytotoxicity was assessed using the tetrazolium bromide-based assay against human colorectal adenocarcinoma (CaCO-2) cell lines. Flocculation efficiencies and mechanisms were determined using Jar and zeta potential assays, respectively. The bioflocculant from K. pneumoniae (Kp1) revealed a fibrous morphology, whereas that from M. guilliermondii (Mg1) displayed a granular structure. FTIR spectra revealed hydroxyl, amine, and alkene groups as key functional groups, while TGA analysis indicated that Kp1 was thermally unstable, contrary to Mg1, which exhibited good thermal stability. Bioflocculants Kp1 and Mg1 exhibited COD removal of 90.86% and 93.12% and turbidity reductions of 92.65% and 92.74%, respectively. Zeta potential analysis revealed that bioflocculant Kp1 primarily flocculated through charge neutralisation, while Mg1 employed a bridging mechanism. These bioflocculants illustrated strong potential to treat wastewater. However, the observed cytotoxic effect at increased concentrations warrants cautious handling and application in lower doses. Full article

Bacteria are promising sources of bioflocculants, yet their regulatory machinery for bioflocculant synthesis remains underexplored. This study focused on evaluating the biosynthetic genes, optimisation and assessment of bioflocculation efficiency in wastewater. The isolated bioflocculant producers were identified by 16S rRNA and rpoB gene analysis. Polymerase chain reaction was used to assess the presence of polyketide synthase I (PKS-1), polyketide synthase II (PKS-II), non-ribosomal peptide synthetase (NRPS), epsH and epsJ. A one-factor-at-a-time technique was utilised for optimisation of culture conditions. The bioflocculants’ efficiencies were evaluated in wastewater using the Jar test method. Among 31 isolates, Klebsiella michiganensis and Klebsiella pasteurii were the most potent bioflocculant producers. They both revealed the presence of PKS-II. K. pasteurii possessed the epsH gene. The optimal conditions for maximum bioflocculant production (95% activity) by K. michiganensis were a temperature of 35 °C, pH of 5, galactose, tryptophan and 84 h of incubation. K. pasteurii’s maximum bioflocculant production of 83% was obtained at a temperature of 35 °C and pH of 7, with galactose, a mixture of urea, yeast extract, and ammonium sulphate (NH₄)₂SO₄ and 96 h of fermentation. Their bioflocculants reduced the chemical oxygen demand and turbidity of wastewater by more than 70%. The bacteria had promising bioflocculant production with potential applicability in wastewater treatment. Full article

Microalgae cultivation is often limited by the high cost and inefficiency of harvesting, making it a major barrier to large-scale resource recovery. Traditional methods such as centrifugation or chemical flocculation are expensive and environmentally unsustainable. As a biological alternative, filamentous fungi can form pellets or hyphal networks that entrap microalgae cells via bio-flocculation, facilitating easier separation from the culture medium. This study aimed to optimize culture conditions for fungal pellet formation and develop effective microalgae–fungal composites for improved harvesting. Fungal isolates including Aspergillus niger, A. fumigatus, A. flavus, and unidentified strains were screened for their pelleting capacity and nutrient removal efficiency. Through OVAT analysis, key factors such as pH, agitation speed, carbon source, and C:N ratio were found to significantly influence pellet formation. One isolate (FP) showed strong potential, forming pellets under stationary conditions and performing best at neutral to alkaline pH. Molecular analysis identified FP as a fungal consortium containing members of Ascomycota, Mucoromycota, Basidiomycota, and Rozellomycota. When applied to cocoa pulp wastewater, the microalgae–fungi composites outperformed monocultures in reducing NH₄⁺-N, COD, and PO₄³⁻-P. Interestingly, microalgae delayed fungal sporulation. These findings highlight the potential of fungal consortia in enhancing both microalgae harvesting and wastewater bioremediation. Full article

Flocculants are indispensable in water and wastewater treatment, enabling the aggregation and removal of suspended particles, colloids, and emulsions. However, the conventional development and application of flocculants rely heavily on empirical methods, which are time-consuming, resource-intensive, and environmentally problematic due to issues such as sludge production and chemical residues. Recent advances in machine learning (ML) have opened transformative avenues for the design, optimization, and intelligent application of flocculants. This review systematically examines the integration of ML into flocculant research, covering algorithmic approaches, data-driven structure–property modeling, high-throughput formulation screening, and smart process control. ML models—including random forests, neural networks, and Gaussian processes—have successfully predicted flocculation performance, guided synthesis optimization, and enabled real-time dosing control. Applications extend to both synthetic and bioflocculants, with ML facilitating strain engineering, fermentation yield prediction, and polymer degradability assessments. Furthermore, the convergence of ML with IoT, digital twins, and life cycle assessment tools has accelerated the transition toward sustainable, adaptive, and low-impact treatment technologies. Despite its potential, challenges remain in data standardization, model interpretability, and real-world implementation. This review concludes by outlining strategic pathways for future research, including the development of open datasets, hybrid physics–ML frameworks, and interdisciplinary collaborations. By leveraging ML, the next generation of flocculant systems can be more effective, environmentally benign, and intelligently controlled, contributing to global water sustainability goals. Full article

Phycosphere niches host rich, unique microbial consortia that harbor complex algae–bacteria interactions with fundamental significance in underpinning most functions of aquatic ecological processes. Therefore, harvesting the uncultured phycobacteria is crucial for understanding the intricate mechanisms governing these dynamic interactions. Here, we characterized and compared microbial community composition of the phycosphere microbiota from six harmful algal bloom-forming marine dinoflagellates, Alexandrium spp., and their bacterial associations. Furthermore, based on a combinational enhanced cultivation strategy (CECS) procedure for the selected isolation for cultivable phycobacteria, a new yellow-pigmented bioactive bacterium designated ABI-6-9 was successfully recovered from cultivable phycobacteria of the highly toxic A. minutum strain amtk4. An additional phylogenomic analysis fully identified this new isolate as a potential novel species of the genus Mameliella within the family Roseobacteraceae. The bioactivity evaluation observed that strain ABI-6-9 can significantly promote the cell growth of its algal host and altered the gonyautoxin accumulation profiles in the co-culture circumstance. Additionally, the bacterial production of active bioflocculanting exopolysaccharides (EPSs) by strain ABI-6-9 was also measured after culture optimization. Thus, these findings revealed the potential environmental and biotechnological implications of this new microalgae growth- promoting phycobacterium. Full article

The growing demand for the sustainable production of high-value compounds, such as biofuels, lipids, and pigments like carotenoids and phycobilin, has become the subject of numerous investigations. Furthermore, this has led to the exploration of renewable methods utilizing microalgae as feedstock to mitigate the challenges associated with producing these valuable compounds. Nevertheless, despite the numerous advantages of microalgae, the development of a microalgal biorefinery that employs sustainable, environmentally friendly, and economically efficient technologies remains a necessity. To address this challenge, the bio-flocculation process, and more specifically self-flocculation, is presented as a cost-effective and energy-efficient solution. This method is as easy and effective as chemical flocculation, which is applied at an industrial scale; however, in contrast, it is sustainable and cost-effective as no costs are involved in the pre-treatment of the biomass for oil extraction or in the pre-treatment of the medium before it can be re-used. In addition, microalgae possess molecular tools that would allow the efficiency of these processes to be increased. In the present review, we summarize the microalgal harvesting technologies used, with a particular focus on bio- and self-flocculation processes, and identify the improvements that could be made to enhance the production of high-added-value compounds while simultaneously reducing costs in microalgae biorefineries. Full article

This study systematically compared the harvesting efficiency and flocculation mechanisms of a bioflocculant (chitosan) and a chemical flocculant (Al₂(SO₄)₃) for Chlorella cells. For the first time, the divergent mechanisms underlying floc structure formation between the two flocculants were elucidated by analyzing the EPS distribution and dynamic changes in microalgal surface functional groups. By optimizing critical operational parameters—including flocculant dosage, flocculation time, pH, and biomass concentration—the optimal dosages of chitosan and Al₂(SO₄)₃ were determined as 0.5 mg/L and 90 mg/L, respectively. Under pH 6, both flocculants achieved over 99% flocculation efficiency within 30 min. Notably, at a dosage of 3 mg/L, chitosan-formed flocs exhibited denser structures, stronger adhesion, and a tendency to aggregate into spherical clusters compared to Al₂(SO₄)₃-induced flocs. Beyond identifying ideal conditions for Chlorella flocculation, this work provides novel insights into the role of EPS and surface functional groups in flocculation mechanisms, offering both theoretical foundations and practical guidance for efficient microalgal harvesting. The findings hold significant implications for optimizing bioflocculant applications and advancing environmentally sustainable harvesting technologies. Full article

Nanotechnology offers effective solutions for removing contaminants and harmful bacteria from polluted water. This study synthesized copper nanoparticles using a carbohydrate-based bioflocculant derived from Proteus mirabilis AB 932526.1. The bioflocculant is a natural polymer that facilitates the aggregation of particles, enhancing the efficiency of the nanoparticle synthesis process. Characterization of the bioflocculant and copper nanoparticles was conducted using Fourier Transform Infrared Spectroscopy, Scanning Electron Microscopy, Energy-Dispersive X-ray Spectroscopy, Ultraviolet-Visible Spectroscopy, X-ray Diffraction, and Transmission Electron Microscopy techniques to assess their properties, flocculation efficiency, and antibacterial characteristics. The optimal flocculation efficiency of 80% was achieved at a copper nanoparticle concentration of 0.4 mg/mL, while a concentration of 1 mg/mL resulted in a lower efficiency of 60%. The effects of biosynthesized copper nanoparticles on human-derived embryonic renal cell cultures were also investigated, demonstrating that they are safe at lower concentrations. The copper nanoparticles effectively removed staining dyes such as safranin (90%), carbol fuchsine (88%), methylene blue (91%), methyl orange (93%), and Congo red (94%), compared to a blank showing only 39% removal. Furthermore, when compared to both chemical flocculants and bioflocculants, the biosynthesized copper nanoparticles exhibited significant nutrient removal efficiencies for nitrogen, sulfur, phosphate, and total nitrates in coal mine and Vulindlela domestic wastewater. Notably, these biosynthesized copper nanoparticles demonstrated exceptional antibacterial activity against both Gram-positive and Gram-negative bacteria. Full article

Spirulina platensis is a promising microalga, but biomass harvesting remains a challenge. Fungal bioflocculation offers a potential solution, facilitating the production of valuable bioproducts like bioethanol. Effective cell disruption methods, including physical-chemical and enzymatic treatments, can enhance biomass utilization. However, commercial enzymes are not optimized for microalgae, necessitating research on ideal operational conditions. This study evaluated physical and enzymatic processes to hydrolyze bioflocculated microalgae for bioethanol production. The microalga was harvested using a fungal bioflocculant produced via submerged fermentation. Biomass hydrolysis involved physical methods (autoclaving, ultrasound + autoclaving, ultrasound + gelatinization, and gelatinization) combined with enzymes (amylase, amyloglucosidase, cellulase, and xylanase), optimized for pH, temperature, and enzyme load. Hydrolysates were then used for bioethanol production. Results showed a microalgae harvest efficiency of 99.7% with a 1:8 fungus-to-microalgae ratio. Enzyme optimization identified ideal conditions (e.g., pH 4.5; 60 °C for amylase/amyloglucosidase, 70 °C for cellulase, and 50 °C for xylanase). Combined enzymatic treatments achieved approximately 70% hydrolysis efficiency, yielding 19.06 g/L glucose and 7.29 g/L ethanol (~79% conversion). Ethanol productivity was ~0.6 g per 1 g bioflocculated biomass L⁻¹·hr. These findings highlight the potential of enzymatic hydrolysis for complex biomasses, although further studies are needed to refine enzyme applications for better biomass utilization. Full article

Phenolic compounds are harmful organic pollutants found in wastewater from the chemical and pharmaceutical industries, which are frequently accompanied by high saline concentrations. Microorganism-based biodegradation represents an environmentally friendly and cost-effective strategy for phenol removal. In this study, we isolated a bioflocculant-producing Bacillus paralicheniformis BL-1 that is capable of phenol degradation in high-salinity conditions. Differential gene expression analysis revealed the down-regulation of genes related to the synthesis of extracellular polymeric substances and the up-regulation of poly-γ-glutamate biosynthesis in 10% NaCl conditions. These findings indicate that poly-γ-glutamate is the main large biomolecule produced by B. paralicheniformis BL-1. A further investigation suggested that salinity stress resulted in the down-regulated expression of the genes involved in iron homeostasis. Therefore, alleviating iron limitation by supplying excess iron could improve cell growth and, thus, increase the phenol removal rate and flocculating activity. The productivity of poly-γ-glutamate reached 2.23 g/L, and the phenol removal rate reached 73.83% in the synthetic medium supplemented with 10% NaCl, 500 mg/L phenol, and 250 μM FeCl₃. Full article

The rising global energy demand and environmental concerns associated with fossil fuels have intensified interest in sustainable biofuel sources, with microalgae emerging as a viable candidate due to its high biomass yield and efficient CO₂ conversion. However, the economic feasibility of microalgal biofuels is currently challenged by costly harvesting processes. This study investigates the use of exopolysaccharides (EPSs) derived from Klebsiella sp. as an environmentally friendly bioflocculant for harvesting two microalgae species: Raphidocelis subcapitata and Dunaliella salina. Comparative flocculation experiments revealed that Klebsiella EPS promotes efficient aggregation in R. subcapitata, achieving over 90% flocculation efficiency, while performance with D. salina was impacted by high salinity, which reduced charge neutralization and bridging effects. Structural analyses using FTIR, 3D-EEM, CLSM, and XPS elucidated the EPS composition, underscoring the roles of polysaccharides and proteins in facilitating microalgal aggregation. The findings indicate that Klebsiella EPS offers a sustainable alternative to chemical flocculants, supporting eco-friendly biofuel production and potential applications in wastewater treatment. This approach provides insights into optimizing EPS-based flocculation for diverse environmental conditions, paving the way for more sustainable biomass recovery practices. Full article

The intriguing characteristics of nanoparticles have fueled recent advancement in the field of nanotechnology. In the current study, a microbial-based bioflocculant made from the SCOBY of Kombucha tea broth was purified, profiled, and utilized to biosynthesize iron nanoparticles as a capping and reducing agent. UV–visible absorption spectroscopy, transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray analysis (EDX), and TGA were used to characterize the Fe nanoparticles. The FT-IR spectra showed functional groups such as hydroxyl, a halogen (C-Br), and carbonyl, and the alkane (C-H) functional groups were present in both samples (bioflocculant and FeNPs) with the exception of the Fe-O bond, which represented the successful biosynthesis of FeNPs. The TEM investigation revealed that the sizes of the produced iron nanoparticles were between 2.6 and 6.2 nm. The UV-vis spectra revealed peaks at 230 nm for the bioflocculant and for the as-fabricated FeNPs, peaks were around 210, 265, and 330 nm, which confirms the formation of FeNPs. X-ray diffraction presented planes (012), (104), (110), (113), (024), (116), and (533) and these planes correspond to 17.17, 32.58, 33.75, 38.18, 45.31, 57.40, and 72.4° at 2Ө. The presence of Fe nanoparticles presented with 0.82 wt% from the EDX spectrum of the biosynthesized FeNPs. However, Fe content was not present from the bioflocculant. SEM images reported cumulus-like particles of the bioflocculant, while that of FeNPs were agglomerated and hexagonal with sizes between 18 and 50 nm. The TGA of FeNPs showed thermal stability by retaining above 60% of its weight at high temperatures. It can therefore be deduced that the purified bioflocculant produced by a yeast Pichia kudraivzevii can be utilized to synthesize FeNPs with the current simple and effective method. Full article

With the aim to establish clean and sustainable sludge treatment, green conditioning using natural flocculants has recently gained a growing interest. In this study, a variety of plant materials, namely Moringa (Moringa oleifera) seeds, Fenugreek (Trigonella foenum-graecum) seeds, Potato (Solanum tuberosum) peels, Aloe (Aloe vera) leaves, Cactus (Opuntia ficus indica) cladodes, and Phragmites (Phragmites australis) stems, were evaluated for their potential bioflocculant activity in conditioning sewage sludge. They were thoroughly characterized to determine their active flocculating compounds. Sludge dewaterability was evaluated by assessing various sludge parameters, including specific resistance to filtration (SRF), dryness of filtration cake (DC), and total suspended solid removal (TSS) from sludge filtrate. The collected results from various physicochemical characterizations of plant materials suggest that the main flocculating agents are carbohydrates in Cactus and Fenugreek and proteins in Moringa, Potato, and Phragmites. Additionally, all tested plant-based flocculants demonstrated effective dewatering performance. Interestingly, compared to the chemical flocculant polyaluminum chloride, Moringa and Cactus showed superior conditioning effects, yielding the lowest SRF values and the highest DC. As a result, the use of these natural flocculants improved sewage sludge filterability, leading to a significant removal of total suspended solids from the filtrate. The conditioning properties of Moringa and Cactus can be attributed to their high protein and sugar content, which facilitates the effective separation of bound water from solids through charge neutralization and bridging mechanisms. Thus, green conditioning using plant-based flocculants, particularly Moringa and Cactus materials, presents a promising and eco-friendly approach to enhance sewage sludge dewatering for safer disposal and valorization. Full article

Ensuring sustainability has increasingly become a significant concern not only in aquaculture but in the general agrifood sector. Therefore, it is imperative to investigate pathways to feed substitutes/best practices to enhance aquaculture sustainability. The application of fungi in aquaculture provides innovative methods to enhance the sustainability and productivity of aquaculture. Fungi play numerous roles in aquaculture, including growth, immunity enhancement and disease resistance. They also play a role in bioremediation of waste and bioflocculation. The application of fungi improves the suitability and utilization of terrestrial plant ingredients in aquaculture by reducing the fibre fractions and anti-nutritional factors and increasing the nutrients and mineral contents of plant ingredients. Fungi are good flotation agents and can enhance the buoyancy of aquafeed. Pigments from fungi enhance the colouration of fish fillets, making them more attractive to consumers. This paper, via the relevant literature, explores the multifaceted roles of fungi in aquaculture, emphasizing their potential to transform aquaculture through environmentally friendly and sustainable techniques. The effectiveness of fungi in reducing fibre fractions and enhancing nutrient availability is influenced by the duration of fermentation and the dosage administered, which may differ for various feed ingredients, making it difficult for most aquaculture farmers to apply fungi approximately. Therefore, the most effective dosage and fermentation duration for each feed ingredient should be investigated. Full article