Microalgae are a promising feedstock for carbon-neutral biofuel production due to their superior cellular composition. Alternatively, oxidative torrefaction has been recognized as a potential thermochemical technique for microalgal solid biofuel upgrading. Herein, by using microalga N. oceanica as a feedstock, several characterizations are adopted for evaluating the potential of oxidative torrefaction towards microalgal solid biofuel production. The oxidatively torrefied microalgae can be upgraded as lignite. After in-depth analysis, significant change in the surface microstructure of oxidatively torrefied microalgae is largely changed (via wrinkle and fragmentation) The hydrophobicity, thermal decomposition, thermal stability, and aromatization of oxidatively torrefied microalgae can be largely enhanced as the oxidative torrefaction severity increase. With the increasing torrefaction temperature, the hydrophobicity of oxidative torrefied microalgae gradually improved. The decomposition of C-2/3/5, and -OCH3, the CO bonds of CH3CO-, and the aromatization occurs via oxidative torrefaction according to the NMR analysis. For XPS analysis, torrefaction operation significantly decreases the carbide carbon and enhances the graphitization. As a result, the thermal stability of oxidatively torrefied microalgae is improved. Conclusively, the information obtained in this study can provide insights into the evaluation of oxidative torrefaction performance and fuel properties of microalgal solid biofuel, which may help accelerate the advancement of oxidative torrefaction industrialization.
In the study, the relationship between the quality and intensity of LED illumination with FAMEs produced were investigated. Nannochloropsis sp. was cultivated for 14 days under different intensities of 100, 150 and 200 μmol photons m(-2) s(-1) of red, blue and mixed red blue LED. The findings revealed that suitable combination of LED wavelengths and intensity; (red LED: 150, blue: 100 and mixed red blue: 200 μmol photons m(-2) s(-1)) produced maximum biomass growth and lipid content. It was observed that the quality and intensity of LED significantly influenced the composition of FAMEs. FAMEs produced under blue LED has high degree of unsaturation (DU) and low cetane number while those under red LED has low DU but higher CN. The combination of red blue LED has produced FAMEs with high ignition and lubricating property and also good oxidation stability indicated by the DU and CN values which lies midway between the red and blue.
Red tides and associated fisheries damage caused by the harmful raphidophyte Chattonella were reassessed based on the documented local records for 50 years to understand the distribution and economic impacts of the harmful species in the Western Pacific. Blooms of Chattonella with fisheries damage have been recorded in East Asia since 1969, whereas they have been only recorded in Southeast Asia since the 1980s. Occurrences of Chattonella have been documented from six Southeast Asian countries, Indonesia, Malaysia, Philippines, Singapore, Thailand and Viet Nam, with mass mortalities mainly of farmed shrimp in 1980-1990s, and farmed fish in 2000-2010s. These occurrences have been reported with the names of C. antiqua, C. marina, C. ovata, C. subsalsa and Chattonella sp., owing to the difficulty of microscopic species identification, and many were not supported with molecular data. To determine the distribution of C. marina complex and C. subsalsa in Southeast Asia, molecular phylogeny and microscopic observation were also carried out for cultures obtained from Indonesia, Malaysia, Japan, Philippines, Russia, Singapore and Thailand. The results revealed that only the genotype of C. marina complex has been detected from East Asia (China, Japan, Korea and Russia), whereas both C. marina complex (Indonesia and Malaysia) and C. subsalsa (Philippines, Singapore and Thailand) were found in Southeast Asia. Ejection of mucocysts has been recognized as a diagnostic character of C. subsalsa, but it was also observed in our cultures of C. marina isolated from Indonesia, Malaysia, Japan, and Russia. Meanwhile, the co-occurrences of the two harmful Chattonella species in Southeast Asia, which are difficult to distinguish solely based on their morphology, suggest the importance of molecular identification of Chattonella genotypes for further understanding of their distribution and negative impacts.
Harvesting microalgae from medium is a major challenge due to their small size and low concentrations. In an attempt to find a cost-effective and eco-friendly harvesting technique, mung bean (Vigna radiata) protein extract (MBPE) was used for flocculation of Nannochloropsis sp. The effects of parameters such as pH, flocculant dose, algae concentration, and mixing time were used to study the flocculation efficiency (FE) of MBPE. Optimum parameters of MBPE dosage of 20 mL L(-1) and a mixing rate of 300 rpm for 6 min achieved a FE of >92% after 2 h of settling time. MBPE-aggregated microlga flocs were characterized by microscopy. Zeta potential values decreased with increasing flocculant dose, and the values obtained were -6.93 ± 0.60, -5.36 ± 0.64, and -4.44 ± 0.22 for doses of 10, 20, and 30 mL L(-1), respectively. In conclusion, MBPE flocculants used in this study are safe, nontoxic, and pollution free, so they could be used for an effective, convenient, and rapid harvesting of microalgae in an eco-friendly approach. These methods are sustainable and could be applied in industrial scale for aquaculture nutrition.
Docosahexaenoic acid (DHA, C22:6n-3) plays a vital role in the enhancement of human health, particularly for cognitive, neurological, and visual functions. Marine microalgae, such as members of the genus Aurantiochytrium, are rich in DHA and represent a promising source of omega-3 fatty acids. In this study, levels of glucose, yeast extract, sodium glutamate and sea salt were optimized for enhanced lipid and DHA production by a Malaysian isolate of thraustochytrid, Aurantiochytrium sp. SW1, using response surface methodology (RSM). The optimized medium contained 60 g/L glucose, 2 g/L yeast extract, 24 g/L sodium glutamate and 6 g/L sea salt. This combination produced 17.8 g/L biomass containing 53.9% lipid (9.6 g/L) which contained 44.07% DHA (4.23 g/L). The optimized medium was used in a scale-up run, where a 5 L bench-top bioreactor was employed to verify the applicability of the medium at larger scale. This produced 24.46 g/L biomass containing 38.43% lipid (9.4 g/L), of which 47.87% was DHA (4.5 g/L). The total amount of DHA produced was 25% higher than that produced in the original medium prior to optimization. This result suggests that Aurantiochytrium sp. SW1 could be developed for industrial application as a commercial DHA-producing microorganism.
In the present study, a novel oleaginous Thraustochytrid containing a high content of docosahexaenoic acid (DHA) was isolated from a mangrove ecosystem in Malaysia. The strain identified as an Aurantiochytrium sp. by 18S rRNA sequencing and named KRS101 used various carbon and nitrogen sources, indicating metabolic versatility. Optimal culture conditions, thus maximizing cell growth, and high levels of lipid and DHA production, were attained using glucose (60 g l⁻¹) as carbon source, corn steep solid (10 g l⁻¹) as nitrogen source, and sea salt (15 g l⁻¹). The highest biomass, lipid, and DHA production of KRS101 upon fed-batch fermentation were 50.2 g l⁻¹ (16.7 g l⁻¹ day⁻¹), 21.8 g l⁻¹ (44% DCW), and 8.8 g l⁻¹ (40% TFA), respectively. Similar values were obtained when a cheap substrate like molasses, rather than glucose, was used as the carbon source (DCW of 52.44 g l⁻¹, lipid and DHA levels of 20.2 and 8.83 g l⁻¹, respectively), indicating that production of microbial oils containing high levels of DHA can be produced economically when the novel strain is used.
Thraustochytrids are getting increasingly popular due to their high potential role as alternative producers of the high-valued ω-3 polyunsaturated fatty acids (PUFA), docosahexaenoic acid (DHA). While most thraustochytrids prefer glucose as the major carbon source, few strains have been reported to prefer fructose. One such strain is Aurantiochytrium sp. SW1. In this study, the effect of fructose on DHA accumulation by SW1 was investigated using a two-level full factorial design. Besides, biomass, lipid and DHA accumulation profiles of SW1 cultivated in fructose and glucose media were compared. Results revealed that fructose has a very significant positive effect on the volumetric DHA content. Meanwhile, its involvement in affecting DHA biosynthetic capacity, though significant, is not very profound. It was also found that when cultivated in fructose medium, SW1 had a less steep log phase compared to that of glucose medium. However, after 48h of cultivation, biomass and lipid accumulation in fructose medium outweighed the other. Volumetric DHA content in fructose medium at 96h was 11% higher than that of glucose medium. Overall, fructose was found to be a more suitable substrate for biomass, lipid and DHA accumulation in SW1 compared to the conventional source, glucose.
Two experiments were conducted to develop Acartia erythraea and Oithona brevicornis cultures: The performance of five microalgal diets to produce nauplii, copepodites and adults of A. erythraea; and the performance of the same diets to produce nauplii, copepodites and adults of O. brevicornis. The five different microalgal diets were Isochrysis galbana (IG), Chaetoceros affinis (CA), Chlorella marina (CM), Nannochloropsis oculata (NO) and mixed algae (mixture of IG, CA, CM and NO at an equal abundance to provide the exact cell density). The results indicated that the abundance of both A. erythraea and O. brevicornis was higher in tanks supplied with IG and mixed algae than the tanks supplied with CA, CM and NO. IG and mixed algal diets were statistically similar on the mean abundance for both A. erythraea and O. brevicornis. The maximum production of A. erythraea nauplii was observed on day 12 of culture period and the nauplii production decreased from day 13 onwards. The mean abundance of A. erythraea copepodites and adults increased along with time up to the end of the culture period. In the case of O. brevicornis nauplii, the maximum abundance was observed on day 9 day of culture period and the nauplii production decreased from day 10 onwards. The mean abundance of O. brevicornis copepodites and adults increased gradually from the beginning to the end of the culture period. Under the experimental conditions of this study, both IG and mixed algal diets can be recommended for the best growth performance of A. erythraea and O. brevicornis.
In this study, Nannochloropsis oceanica CY2 was grown in deep-sea water (DSW)-based medium in 5-L plastic bag-type photobioreactors (PBRs) for the autotrophic production of Eicosapentaenoic acid (EPA, 20:5n-3). EPA production of N. oceanica CY2 was stimulated when it was grown in 100% DSW amended with 1.5 g L-1 NaNO3, achieving a EPA content of 3.1% and a biomass concentration of 3.3 g L-1. An outdoor-simulated microalgae cultivation system was also conducted to validate the feasibility of outdoor cultivation of the CY2 strain in plastic bag-type PBRs. Using an inoculum size of 0.6 g/L, the biomass concentration in the PBR culture was 3.5 g L-1, while the EPA content and productivity reached a maximal level of 4.12% and 7.49 mg L-1 d-1, respectively. When the PBRs were operated on semi-batch mode, the EPA productivity could further increase to 9.9 mg L-1 d-1 with a stable EPA content of 4.1%.
The types of microalgae strains and the method used in lipid extraction have become crucial factors which influence the productivity of crude oil. In this paper, Nannochloropsis sp. and Tetraselmis sp. were chosen as the strains and four different methods were used to extract the lipids: Hara and Radin, Folch, Chen and Bligh and Dyer. These methods were performed by using conventional heating and microwave irradiation methods. Results revealed that highest lipid yield from the different species was obtained using different extraction methods; both under microwave irradiation. The lipid yield for Tetraselmis sp. and Nannochloropsis sp. was highest when Hara and Radin (8.19%), and Folch (8.47%) methods were used respectively under microwave irradiation. The lipids extracted were then transesterified to biodiesel and the quality of the biodiesel was analyzed using the gas chromatography.
The use of acetosyringone in Agrobacterium-mediated gene transfer into plant hosts has been favored for the past few decades. The influence of other phenolic compounds and their effectiveness in Agrobacterium-mediated plant transformation systems has been neglected. In this study, the efficacy of four phenolic compounds on Agrobacterium-mediated transformation of the unicellular green alga Nannochloropsis sp. (Strain UMT-M3) was assessed by using β-glucuronidase (GUS) assay. We found that cinnamic acid, vanillin and coumarin produced higher percentages of GUS positive cells as compared to acetosyringone. These results also show that the presence of methoxy group in the phenolic compounds may not be necessary for Agrobacterium vir gene induction and receptor binding as suggested by previous studies. These findings provide possible alternative Agrobacterium vir gene inducers that are more potent as compared to the commonly used acetosyringone in achieving high efficiency of Agrobacterium-mediated transformation in microalgae and possibly for other plants.
Natural growth-promoting nutrients extracted from aquaculture sludge waste can be used to maximise microalgal growth. This study identified the influence of aquaculture sludge extract (SE) on four microalgae species. Conway or Bold's Basal Media (BBM) was supplemented with SE collected from a Sabak Bernam shrimp pond (SB) and Kota Puteri fish pond (KP), and tested using a novel microplate-incubation technique. Five different autoclave extraction treatment parameters were assessed for both collected SE, i.e., 1-h at 105 °C, 2-h at 105 °C, 1-h at 121 °C, 2-h at 121 °C, and 24-h at room temperature (natural extraction). Microalgae culture in the microplates containing control (media) and enriched (media + SE) samples were incubated for nine days, at 25 °C with the light intensity of 33.75 μmol photons m-2 s-1 at 12-h light/dark cycle. The total dissolved nitrogen (TDN) and total dissolved phosphorus (TDP) in KP SE were 44.0-82.0 mg L-1 and 0.96-8.60 mg L-1. TDN (8.0%-515.0%) and TDP (105%-186 %) were relatively higher in KP SE compared to SB SE. The growth of microalgae species Nannochloropsis ocenica showed significant differences (p < 0.05) between the five extraction treatments from SB and the control. However, Chlorella vulgaris, Neochloris conjuncta, and Nephroclamys subsolitaria showed no significant differences (p > 0.05) in SB SE. N. ocenica, C. vulgaris, and N. conjuncta showed significant differences (p < 0.05) between five extraction treatments from KP and the control while N. subsolitaria showed no significant difference (p > 0.05). The specific growth rate (SGR) in the exponential phase of all microalgae species were relatively higher in SB SE compared to KP SE. While the organic matter content of KP SE was relatively higher, there were no significant differences in microalgae growth compared to SB SE. Nonetheless, modified SE did influence microalgae growth compared to the control. This study shows that modified SE could be used as enrichment media for microalgae cultivation.
Microalgae can produce various natural products such as pigments, enzymes, unique fatty acids and vitamin that benefit humans. The objective of the study is to study the bioaccessibility of carotenoids (β-carotene and lycopene) and vitamin E (α- and β-tocopherol) of Nannochloropsis oculata and Chaetoceros calcitrans. Analyses were carried out for both the powdered forms of N. oculata and C. calcitrans, and the dried extract forms of N. oculata and C. calcitrans. In vitro digestion method together with RP-HPLC was used to determine the bioaccessibility of carotenoids and vitamin E for both forms of microalgae. Powdered form of N. oculata had the highest bioaccessibility of β-carotene (28.0 ± 0.6 g kg-1), followed by dried extract N. oculata (21.5 ± 1.1 g kg-1), dried extract C. calcitrans (16.9 ± 0.1 g kg-1), and powdered C. calcitrans (15.6 ± 0.1 g kg-1). For lycopene, dried extract of N. oculata had the highest bioaccessibility of lycopene (42.6 ± 1.1 g kg-1), followed by dried extract C. calcitrans (41.9 ± 0.6 g kg-1), powdered C. calcitrans (39.7 ± 0.1 g kg-1) and powdered N. oculata (32.6 ± 0.7 g kg-1). Dried extract C. calcitrans had the highest bioaccessibility of α-tocopherol (72.1 ± 1.2 g kg-1). However, β-tocopherol was not detected in both dried extract and powdered form of C. calcitrans. In conclusion, all samples in their dried extract forms were found to have significantly higher bioaccessibilities than their powdered forms. This may be due to the disruption of the food matrix contributing to a higher bioaccessibility of nutrients shown by the dried extract forms.
Microalgae biorefinery is presently receiving a lot of attention as driven by its production of high value-added products. In this study, an oleaginous microalga Aurantiochytrium limacinum SR21 was cultured for docosahexaenoic acid (DHA) production using 20% (w/v) of K2HPO4-waste feedstock to replace 0.005% (w/v) of KH2PO4 in the flask culture. DHA is an essential nutrient for human's brain functionalities. Collectively, the K2HPO4-waste feedstock with working concentration of 0.005% (w/v) in the cultivation prompted a higher lipid content (8.29%) and DHA production (128.81 mg.L-1). Moreover, natural plant pigment products containing stabilised betacyanins were utilised as natural red colourants for hard candy production. This study develops microalgal cultivation using salt-rich waste feedstock for a higher lipid and DHA content as well as application of natural colouring agents in food products.
This study was conducted to investigate and compare the fatty acids and tocopherols of lipid extracted from marine microalgae, Nannochloropsis oculata (NO) and Tetraselmis suecica (TS) using solvent extraction and supercritical fluid extraction (SFE). Fatty acids and tocopherols were determined in the extracted lipid as functions of the temperature (40, 80oC) and pressure (3000, 5000, 7000, 9000psi). Dichloromethane/methanol and hexane were the chosen conventional solvent for fatty acids and tocopherols extraction respectively. The results obtained showed that there were differences in the fatty acid composition of various lipid extracts of NO and TS. Extracts of NO were high in myristic acid (C14:0) (17-35%), palmitic acid (C16:0) (14-47%) and palmitoleic acid (C16:1) (11-42%) whereas extracts of TS were high in C14:0 (21-34%) and C16:0 (29-49%). Eicosapentaenoic acid (EPA) was detected only under certain SFE conditions in NO but was not detected in TS. α-, β- and γ-tocopherol were detected in various SFE extracts of NO but only α- and β-tocopherol were detected in TS. Hexane extraction of both NO and TS resulted in the detection of only α-tocopherol. In conclusion, the use of different extraction methods resulted in different compositions and concentrations of fatty acids and tocopherols in the microalgae studied.
Intestinal protozoa are increasingly being studied because of their association with acute and chronic diarrhoea in immunocompromised as well as immunocompetent patients. Various community outbreaks due to contamination of water or food with these protozoa have further highlighted their importance in public health. Among these important pathogens are Giardia duodenalis, Entamoeba histolytica, Cryptosporidium parvum, Cyclospora cayetanensis, Isospora belli, and microsporidia. Except for the cyst-forming G. duodenalis and E. histolytica, the others are intracellular and form spores which are passed out with the faeces. These organisms are also found in various animals and birds and zoonotic transmission is thought to occur. These infections are distributed worldwide, with a higher prevalence in developing compared to developed countries. However, the relative importance of zoonotic infections especially in developing countries has not been studied in detail. The prevalence rates are generally higher in immunodeficient compared to immunocompetent patients. Higher prevalence rates are also seen in rural compared to urban communities. Most studies on prevalence have been carried out in developed countries where the laboratory and other health infrastructure are more accessible than those in developing countries. This relative inadequacy of laboratory diagnosis can affect accurate estimates of the prevalence of these infections in developing countries. However, reports of these infections in travellers and workers returning from developing countries can provide some indication of the extent of these problems. Most studies on prevalence of amoebiasis in developing countries were based on morphological identification of the parasite in faecal smears. As the pathogenic E. histolytica is morphologically indistinguishable from that of non-pathogenic E. dispar, estimates of amoebiasis may not be accurate. The epidemiology of human microsporidia infections is not completely understood. Two species, Enterocytozoon bieneusi and Encephalitozoon intestinalis, are associated with gastrointestinal disease in humans and it is believed that human to human as well as animal to human infections occur. However, the importance of zoonotic infections has not been fully characterised. G. duodenalis cysts, microsporidia and Cryptosporidium oocysts have been detected in various ground water resources, but their role in community outbreaks and maintenance of the infection has not been fully characterised. The taxonomic classification and pathogenic potential of B. hominis are still controversial. While considered by many as yeast, fungi or protozoon, recent sequence analysis of the complete SSUrRNA gene has placed it within an informal group, the stramenopiles. This review covers recent published data on these zoonotic infections and examines their public health importance in Asian countries.