Inorganic and synthetic flocculants are widely investigated for removing harmful microalgae, such as Microcystis aeruginosa. However, their toxicity and non-biodegradability are shortcomings. Bioflocculants based on extracellular polysaccharides have attracted much attention as alternative flocculants. However, its high production cost is a limiting factor for applying bioflocculants. Here, we investigate the potential of the dead cells of a marine filamentous bacterium, Aureispira sp. CCB-QB1, as a novel flocculant on M. aeruginosa cells. The removal efficiency of M. aeruginosa cells by the dead cells was measured by mixing and shaking both components in a buffer with 5 mM CaCl2 in different incubation times and concentrations of the dead cells. After that, the minimum effective concentration of CaCl2 was determined. The combination effect of FeCl3 and the dead cells on the removal efficiency was tested. The structure of cell aggregates consisted of the dead cells and M. aeruginosa cells were also observed using a scanning electron microscope. The maximum removal efficiency (75.39%) was reached within 3 min in the presence of CaCl2 when 5 mg/ml of the dead cells (wet cells) were added. The optimal concentration of CaCl2 was 5 mM. The combination of the dead cells and a low concentration of FeCl3 (10 mg/L) with 5 mM of CaCl2 significantly improved the removal efficiency by about 1.2 times (P
Cesium-137 (137Cs) is one of the radioactive substances that was released into the environment as a result of the Fukushima nuclear disaster. Radiocesium exposure is of great concern due to its potential environmental implications. However, research on 137Cs removal using algae is still limited. This is the first report to describe the kinetic properties of 137Cs uptake by Vacuoliviride crystalliferum in the presence and absence of potassium. In this work, we studied the kinetic properties of 137Cs uptake using a freshwater microalga, V. crystalliferum (NIES 2860). We also analyzed the effects of temperature, light, and potassium (K) on the 137Cs uptake. Results showed that V. crystalliferum can remove up to 90% of 157 nM 137Cs within an hour. At 20 °C, the removal increased by up to 96%, compared to less than 10% at 5 °C. However, the removal was inhibited by nearly 90% in the dark compared to the removal in the light, implying that V. crystalliferum cells require energy to accumulate 137Cs. In the inhibition assay, K concentrations ranged from 0 to 500 µM and the inhibitory constant (Ki) for K was determined to be 16.7 µM. While in the uptake assay without potassium (- K), the Michaelis constant (Km) for Cs was 45 nM and increased to 283 nM by the addition of 20 µM potassium (+ K), indicating that V. crystalliferum had a high affinity for 137Cs. In addition, the maximum uptake velocity (Vmax) also increased from 6.75 to 21.10 nmol (mg Chl h)-1, implying the existence of Cs active transport system. In conclusion, V. crystalliferum is capable of removing radioactive 137Cs from the environment and the removal was favorable at both normal temperature and in the light.
Cultivation of microalgae in wastewater stream has been extensively reported, especially for simultaneous production of biolipid and wastewater treatment process. This study aimed to derive the research trend and focus on biolipid production from microalgae cultivated in wastewater by using bibliometric approach. The search strategy used in Scopus database resulted in 1339 research articles from 1990 to November 2023. Majority of publications (46%) were affiliated to China and India, showing their predominance in this field. Keywords related to the center of attention included biodiesel, biofuel, biomass and nutrient removal. Meanwhile, keyword with recent publication year, indicating the emerging research trends, revolved around the cultivation techniques and application of the system. Co-culture involving more than one microalgae species, bacteria and yeast showed promising results, while addition of nanoparticles was also found to be beneficial. Increasing exploration on the application of microalgae for treatment of saline wastewater was also reported and the carbon fixation mechanism by microalgae has been widely investigated to promote less environmental impact. Future research on these topics were suggested based on the findings of the bibliometric analyses.
Microalgae are important microorganisms which produce potentially valuable compounds. Astaxanthin, a group of xanthophyll carotenoids, is one of the most powerful antioxidants mainly found in microalgae, yeasts, and crustaceans. Environmental stresses such as intense light, drought, high salinity, nutrient depletion, and high temperature can induce the accumulation of astaxanthin. Thus, this research aims to investigate the effect of black light, also known as long-wave ultraviolet radiation or UV-A, as a stressor on the accumulation of astaxanthin as well as to screen the antioxidant property in two tropical green algal strains isolated from Malaysia, Coelastrum sp. and Monoraphidium sp. SP03. Monoraphidium sp. SP03 showed a higher growth rate (0.66 day-1) compared to that of Coelastrum sp. (0.22 day-1). Coelastrum sp. showed significantly higher accumulation of astaxanthin in black light (0.999 g mL culture-1) compared to that in control condition (0.185 g mL-1). Similarly, Monoraphidium sp. SP03 showed higher astaxanthin content in black light (0.476 g mL culture-1) compared to that in control condition (0.363 g mL culture-1). Coelastrum sp. showed higher scavenging activity (30.19%) when cultured in black light condition, indicating a correlation between the antioxidant activity and accumulation of astaxanthin. In this study, black light was shown to possess great potential to enhance the production of astaxanthin in microalgae.
The advantageous characteristics of aerobic granular sludge (AGS) have led to their increasing popularities among academics and industrial players. However, there has been no bibliometric report on current and future research trends of AGS. This study utilized the available reports of AGS in the Scopus database for comprehensive bibliometric analyses using VOSviewer software. A total of 1203 research articles from 1997 to 2020 were analyzed. The dominance of the Netherlands and China were revealed by the high number of publications and citations. Nevertheless, the Netherlands exhibited higher average citation per article at 76.4. A recent process of AGS involving biochar and algal addition were also identified. Meanwhile, the application of AGS for antibiotic containing wastewater as well as possibility of resource recovery were recently reported and was expected to expand in the future. It was suggested that application of AGS would develop further along with the development of sustainable wastewater treatment process.
Despite various research works on algal-bacterial aerobic granular sludge for wastewater treatment and resource recovery processes, limited information is available on its application in real wastewater treatment in terms of performance, microbial community variation and resource recovery. This study investigated the performance of algal-bacterial aerobic granular sludge on real low-strength wastewater treatment in addition to the characterization of microbial community and fatty acid compositions for biodiesel production. The results demonstrated 71% COD, 77% NH4+-N and 31% phosphate removal efficiencies, respectively. In addition, all the water parameters successfully met the effluent standard A, imposed by the Department of Environment (DOE) Malaysia. Core microbiome analyses revealed important microbial groups (i.e., Haliangium ochraceum, Burkholderiales and Chitinophagaceae) in bacterial community. Meanwhile the photosynthetic microorganisms, such as Oxyphotobacteria and Trebouxiophyceae dominated the algal-bacterial aerobic granular sludge, suggesting their important roles in granulation and wastewater treatment. Up to 12.51 mg/gSS lipid content was recovered from the granules. In addition, fatty acids composition showed high percetages of C16:0 and C18:0, demonstrating high feasibility to be used for biodiesel production application indicated by the cetane number, iodine value and oxidation stability properties.
Limited information is available on the characteristics of algal-bacterial aerobic granular sludge (AGS) treating real wastewater, especially on its alginate-like exopolymers (ALE) production. In addition, the effect of target microalgae species inoculation on the system performance has not been fully understood. This study aimed to reveal the effect of microalgae inoculation on the characteristics of algal-bacterial AGS and its ALE production potential. Two photo-sequencing batch reactors (PSBR) were employed, namely R1 with activated sludge and R2 with Tetradesmus sp. and activated sludge being inoculated, respectively. Both reactors were fed with locally sourced municipal wastewater and operated for 90 days. Algal-bacterial AGS were successfully cultivated in both reactors. No significant difference was observed between the performances of R1 and R2, reflecting that the inoculation of target microalgae species may not be crucial for the development of algal-bacterial AGS when treating real wastewater. Both reactors achieved an ALE yield of about 70 mg/g of volatile suspended solids (VSS), indicating that a substantial amount of biopolymer can be recovered from wastewater. Interestingly, boron was detected in all the ALE samples, which might contribute to granulation and interspecies quorum sensing. The enrichment of lipids content in ALE from algal-bacterial AGS treating real wastewater reveals its high resource recovery potential. Overall, the algal-bacterial AGS system is a promising biotechnology for simultaneous municipal wastewater treatment and resource (like ALE) recovery.
Tetraselmis chuii is a potential microalgae that is in consideration for producing bioethanol owing to its large content of carbohydrates. The glucose production from T. chuii through an enzymatic process with cellulase and xylanase (pretreatment process) and α-amylase and glucoamylase (saccharification process) was studied. The mechanism of the enzymatic process was developed and the kinetic models were then evaluated. For the pretreatment process, enzymes with 30% concentration reacted at 30 °C for 40 min resulted in 35.9% glucose yield. For the saccharification process, the highest glucose yield of 90.03% was obtained using simultaneous α-amylase (0.0006%) and glucoamylase (0.01%) enzymes at 55 °C and for 40 min. The kinetic models fitted well with the experimental data. The model also revealed that the saccharification process performed better than the pretreatment process with a higher kinetic constant and lower activation energy. The proposed kinetic model plays an important role in implementing processes at a larger scale.
The emergence of antibiotic resistance among multidrug-resistant (MDR) microbes is of growing concern, and threatens public health globally. A total of 129 Escherichia coli isolates were recovered from lowland aqueous environments near hospitals and medical service centers in the vicinity of Kuala Lumpur, Malaysia. Among the eleven antibacterial agents tested, the isolates were highly resistant to trimethoprim-sulfamethoxazole (83.7%) and nalidixic acid (71.3%) and moderately resistant to ampicillin and chloramphenicol (66.7%), tetracycline (65.1%), fosfomycin (57.4%), cefotaxime (57.4%), and ciprofloxacin (57.4%), while low resistance levels were found with aminoglycosides (kanamycin, 22.5%; gentamicin, 21.7%). The presence of relevant resistance determinants was evaluated, and the genotypic resistance determinants were as follows: sulfonamides (sulI, sulII, and sulIII), trimethoprim (dfrA1 and dfrA5), quinolones (qnrS), β-lactams (ampC and blaCTX-M), chloramphenicol (cmlA1 and cat2), tetracycline (tetA and tetM), fosfomycin (fosA and fosA3), and aminoglycosides (aphA1 and aacC2). Our data suggest that multidrug-resistant E. coli strains are ubiquitous in the aquatic systems of tropical countries and indicate that hospital wastewater may contribute to this phenomenon.
Antibiotic resistance has become a major public health problem throughout the world. The presence of antibiotic-resistant bacteria such as Staphylococcus aureus and antibiotic resistance genes (ARGs) in hospital wastewater is a cause for great concern today. In this study, 276 Staph. aureus isolates were recovered from hospital wastewater samples in Malaysia. All of the isolates were screened for susceptibility to nine different classes of antibiotics: ampicillin, ciprofloxacin, gentamicin, kanamycin, erythromycin, vancomycin, trimethoprim and sulfamethoxazole, chloramphenicol, tetracycline and nalidixic acid. Screening tests showed that 100 % of Staph.aureus isolates exhibited resistance against kanamycin, vancomycin, trimethoprim and sulfamethoxazole and nalidixic acid. Additionally, 91, 87, 50, 43, 11 and 8.7 % of isolates showed resistance against erythromycin, gentamicin, ciprofloxacin, ampicillin, chloramphenicol and tetracycline, respectively. Based on these results, 100 % of isolates demonstrated multidrug-resistant (MDR) characteristics, displaying resistance against more than three classes of antibiotics. Of 276 isolates, nine exhibited resistance to more than nine classes of tested antibiotics; these were selected for antibiotic susceptibility testing and examined for the presence of conserved ARGs. Interestingly, a high percentage of the selected MDR Staph.aureus isolates did not contain conserved ARGs. These results indicate that non-conserved MDR gene elements may have already spread into the environment in the tropics of Southeast Asia, and unique resistance mechanisms against several antibiotics may have evolved due to stable, moderate temperatures that support growth of bacteria throughout the year.
Natural astaxanthin is known to be produced by green microalgae, a potent producer of the most powerful antioxidant. To increase the productivity of astaxanthin in microalgae, random mutagenesis has been extensively used to improve the yield of valuable substances. In the presented work, a newly isolated Coelastrum sp. was randomly mutagenized by exposure to ethyl methane sulfonate and further screened using two approaches; an approach for high growth mutant and an approach for high astaxanthin producing mutant with a high-throughput screening method using glufosinate. Among these, mutant G1-C1 that was selected using glufosinate showed the highest of total carotenoids (45.48±1.5 mg/L) and astaxanthin (28.32±2.5 mg/L) production, which was almost 2-fold higher than that of wild type. This study indicates that random mutagenesis via chemical mutation strategy and screening using glufosinate successfully expedited astaxanthin production in a mutated strain of a Coelastrum sp.
The identification of microalgae species is an important tool in scientific research and commercial application to prevent harmful algae blooms (HABs) and recognizing potential microalgae strains for the bioaccumulation of valuable bioactive ingredients. The aim of this study is to incorporate rapid, high-accuracy, reliable, low-cost, simple, and state-of-the-art identification methods. Thus, increasing the possibility for the development of potential recognition applications, that could identify toxic-producing and valuable microalgae strains. Recently, deep learning (DL) has brought the study of microalgae species identification to a much higher depth of efficiency and accuracy. In doing so, this review paper emphasizes the significance of microalgae identification, and various forms of machine learning algorithms for image classification, followed by image pre-processing techniques, feature extraction, and selection for further classification accuracy. Future prospects over the challenges and improvements of potential DL classification model development, application in microalgae recognition, and image capturing technologies are discussed accordingly.
Geosmin and 2-methylisoborneol (MIB) outbreaks in tropical water bodies, such as Southeast Asia, by actinomycetes have not yet been elucidated in detail. Six Streptomyces isolates from lowland environments in Malaysia were selected and evaluated for their odor production under different temperatures. The gene responsible for the production of geosmin, geoA, was detected in all isolates, while only two isolates harbored tpc, which is responsible for 2-MIB production. This result suggested that geosmin and 2-MIB synthesis pathway genes already existed in the environment in the Tropics of Southeast Asia. Furthermore, our isolates produced musty odor compounds at 30°C, and differences were observed in musty odor production between various temperatures. This result indicated the potential for odor episodes in water bodies of the tropical countries of Southeast Asia throughout the year due to the mean annual ambient temperature of 27°C in the lowlands.
Cadmium (Cd) has become a severe issue in relatively low concentration and attracts expert attention due to its toxicity, accumulation, and biomagnification in living organisms. Cd does not have a biological role and causes serious health issues. Therefore, Cd pollutants should be reduced and removed from the environment. Microalgae have great potential for Cd absorption for waste treatment since they are more environmentally friendly than existing treatment methods and have strong metal sorption selectivity. This study evaluated the tolerance and ability of the microalga Tetratostichococcus sp. P1 to remove Cd ions under acidic conditions and reveal mechanisms based on transcriptomics analysis. The results showed that Tetratostichococcus sp. P1 had a high Cd tolerance that survived under the presence of Cd up to 100 µM, and IC50, the half-maximal inhibitory concentration value, was 57.0 μM, calculated from the change in growth rate based on the chlorophyll content. Long-term Cd exposure affected the algal morphology and photosynthetic pigments of the alga. Tetratostichococcus sp. P1 removed Cd with a maximum uptake of 1.55 mg g-1 dry weight. Transcriptomic analysis revealed the upregulation of the expression of genes related to metal binding, such as metallothionein. Group A, Group B transporters and glutathione, were also found upregulated. While the downregulation of the genes were related to photosynthesis, mitochondria electron transport, ABC-2 transporter, polysaccharide metabolic process, and cell division. This research is the first study on heavy metal bioremediation using Tetratostichococcus sp. P1 and provides a new potential microalga strain for heavy metal removal in wastewater.[Figure: see text]Abbreviations:BP: Biological process; bZIP: Basic Leucine Zipper; CC: Cellular component; ccc1: Ca (II)-sensitive cross complementary 1; Cd: Cadmium; CDF: Cation diffusion facilitator; Chl: Chlorophyll; CTR: Cu TRansporter families; DAGs: Directed acyclic graphs; DEGs: Differentially expressed genes; DVR: Divinyl chlorophyllide, an 8-vinyl-reductase; FPN: FerroportinN; FTIR: Fourier transform infrared; FTR: Fe TRansporter; GO: Gene Ontology; IC50: Growth half maximal inhibitory concentration; ICP: Inductively coupled plasma; MF: molecular function; NRAMPs: Natural resistance-associated aacrophage proteins; OD: Optical density; RPKM: Reads Per Kilobase of Exon Per Million Reads Mapped; VIT1: Vacuolar iron transporter 1 families; ZIPs: Zrt-, Irt-like proteins.