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Enhanced Hydrogen Production by the Halotolerant Cyanobacterium Aphanothece halophytica Through Bacterial Co-Cultivation
(2026-05-01)
Somsin, Chutikarn
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Chinchusak, Nattanon
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Incharoensakdi, Aran
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Phunpruch, Saranya
Hydrogen (H<inf>2</inf>) is a promising clean energy carrier with the potential to partially replace fossil fuels. Biological H<inf>2</inf> production using microorganisms offers an environmentally friendly alternative. The halotolerant cyanobacterium Aphanothece halophytica can produce H<inf>2</inf> under nitrogen-deprived and dark anaerobic conditions. In this study, a co-culture strategy was investigated to enhance H<inf>2</inf> production. Five bacterial strains were screened for their ability to improve H<inf>2</inf> production when co-cultivated with A. halophytica. Among them, Staphylococcus aureus significantly enhanced H<inf>2</inf> production, achieving a maximum rate of 11.11 ± 0.18 µmol H<inf>2</inf> g−1 dry weight h−1. Optimization of the bacterial partner revealed that S. aureus cells harvested at 12 h in the mid-logarithmic phase with an OD<inf>600</inf> of 4.0 were the most effective. An inoculum ratio of A. halophytica to S. aureus of 4:1 further enhanced H<inf>2</inf> production, increased bidirectional hydrogenase activity, and reduced O<inf>2</inf> accumulation. Under optimal conditions (0.945 mmol C-atom L−1 glucose, 0.25 M NaCl, pH 7.4, and 35 °C), the maximum H<inf>2</inf> production rate reached 132.49 ± 4.45 µmol H<inf>2</inf> g−1 dry weight h−1, approximately 5.5-fold higher than that under normal conditions. The co-culture achieved a cumulative H<inf>2</inf> yield of 3248.51 ± 88.11 µmol H<inf>2</inf> g−1 dry weight after 48 h.
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Stable Nanoemulsion System Development Enabling the Topical Delivery of Synechococcus-Derived Peptides
(2026-03-01)
Keawbankrud, Wannisa
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Nakyai, Wongnapa
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Phunpruch, Saranya
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Sangtanoo, Papassara
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Karnchanatat, Aphichart
Marine microalgae including Synechococcus sp. VDW offer the potential to serve as a source of bioactive peptides offering valuable antioxidant and antimelanogenic qualities for use in the pharmaceutical and cosmetics industries. At present, however, the use of such peptides is challenging due to their poor physicochemical stability. This research therefore sought to achieve the production and characterization of a stable nanoemulsion system based upon the use of synthetic Synechococcus-derived peptides through the process of high-pressure homogenization (HPH). Preparation of the nanoemulsions involved the use of Tween-80 and caprylic/capric triglyceride at a pressure of 7,500 psi to perform homogenization for varying durations of 15, 30 and 45 min. Using the optimized formulation with 0.1% w/w peptide for a time of 45 min resulted in 122.16 nm droplets while the zeta potential was −80.09 mV and the PDI (polydispersity index) value was 0.13. Colloidal stability could be considered high, while physical stability under thermal cycling, centrifugation and freeze-thaw cycles was very good, with no phase separation. For all testing intervals, the viscosity and refractive index were stable. It can thus be argues that the HPH approach is suitable to produce peptide-loaded nanoemulsions offering good stability and useful physicochemical characteristics. The developed nanoemulsion system has been optimized to offer potential for applications involving the transdermal delivery of marine peptides in the cosmeceutical sector and for a range of dermal therapies.
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Enhanced hydrogen production by green alga Scenedesmus obliquus TISTR 8546 under atmospheric air through potassium deprivation and cysteine supplementation
(2026-03-01)
Sereetrakul, Kodchaporn
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Taikhao, Samart
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Chinchusak, Nattanon
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Phunpruch, Saranya
Hydrogen is a promising alternative energy carrier that can be produced by green microalgae. However, the key enzyme catalyzing this process, [FeFe]‑hydrogenase, is highly sensitive to O<inf>2</inf> generated during photosynthesis, which severely restricts its activity. This study aimed to screen the microalgal strain capable of producing high levels of H<inf>2</inf> in the presence of O<inf>2</inf> and to enhance H<inf>2</inf> production yields using O<inf>2</inf>-scavenging strategies to levels comparable to or exceeding those achieved under anaerobic conditions. Among the 24 strains tested, Scenedesmus obliquus TISTR 8546 demonstrated the highest H<inf>2</inf> production under both aerobic and anaerobic conditions. The O<inf>2</inf>I<inf>50</inf> of S. obliquus TISTR 8546 cells for H<inf>2</inf> evolution was 15.93 ± 0.24%. Potassium deprivation significantly enhanced H<inf>2</inf> production and hydrogenase activity by lowering O<inf>2</inf> levels through reduced photosynthetic O<inf>2</inf> evolution and increased dark respiration. Moreover, potassium deprivation promoted intracellular starch accumulation, providing reducing equivalents for H<inf>2</inf> generation. Cysteine supplementation further stimulated H<inf>2</inf> production by serving as a reducing agent. S. obliquus TISTR 8546 exhibited a maximum H<inf>2</inf> production rate of 22.92 ± 1.05 μmol H<inf>2</inf> mg Chl−1 h−1 and achieved a maximum cumulative H<inf>2</inf> production of 1153.78 ± 52.65 μmol H<inf>2</inf> mg Chl−1 when incubated in potassium-deprived TAP medium supplemented with 0.1 mM cysteine under atmospheric air for 6 days. This rate was 23.6- and 2.7-fold higher than those obtained in TAP and TAP-K media, respectively. These findings demonstrate the potential of S. obliquus TISTR 8546 as a robust microalgal strain for sustainable H<inf>2</inf> production under atmospheric air, highlighting its promise for future industrial applications.
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Microwave-Assisted Biosynthesis of Silver Nanoparticles Using Chlorella sp. for Antibacterial and Cytotoxicity Effects of Breast Cancer Cell Line
(2026-03-01)
Manklinniam, Piyapan
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Pornroongruengchok, Weerawat
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Phunpruch, Saranya
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Phaepilin, Adisorn
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Pook-In, Grissana
Microwave-assisted biosynthesis using marine Chlorella sp. extracts provides a green and efficient route for the production of silver nanoparticles (AgNPs). Compared with the conventional method (24 h), microwave-assisted synthesis reduces the reaction time to less than 7 min while producing smaller and more uniformly distributed nanoparticles. AgNPs were synthesized using extracts obtained with different solvents and directly compared with those produced via the conventional method to substantiate the efficiency of the microwave-assisted approach. UV–visible spectroscopy confirmed rapid nanoparticle formation, exhibiting surface plasmon resonance peaks in the range of 405 to 427 nm. TEM analysis revealed predominantly spherical AgNPs with particle sizes of approximately 10 to 20 nm. The XRD and FTIR analyses confirmed their crystalline structure and stabilization by algal-derived functional groups. The biological activities of the AgNPs were dependent on the extraction solvent. AgNPs synthesized using hexane extracts exhibited pronounced antibacterial activity, achieving minimum inhibitory concentrations as low as 0.31 µg/mL. In addition, the AgNP induced concentration-dependent cytotoxic effects in human breast cancer cell lines. IC<inf>50</inf> values, determined via dose–response analysis, ranged from 0.18 to 0.67 μg/mL in MDA-MB-231 cells and 1.70 to 8.42 μg/mL in MCF-7 cells. These results indicate a potent cytotoxic profile, with MDA-MB-231 cells exhibiting significantly higher sensitivity to the microwave-assisted formulations. Collectively, these findings highlight microwave-assisted algal-mediated biosynthesis as a sustainable and effective platform for generating bioactive AgNPs with promising antibacterial and anticancer potential.
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Microwave-assisted biosynthesis of silver nanoparticles using two marine microalgal extracts and their antimycobacteriosis activity against bacteria isolated from Betta splendens
(2025-12-01)
Manklinniam, Piyapan
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Phunpruch, Saranya
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Sakulkalavek, Aparporn
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Sakdanuphab, Rachsak
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Worananthakij, Worakrit
This study explores the synthesis and antibacterial properties of silver nanoparticles (AgNPs) as a safer, eco-friendly alternative to traditional chemical treatments for bacterial infections. AgNPs were synthesized using aqueous extracts of marine microalgae, Isochrysis galbana and Chaetoceros calcitrans, via conventional and microwave-assisted methods, with the latter accelerating nanoparticle production. Extracts in ethanol, hexane, and acetone were tested, with the ethanolic extract of I. galbana showing the strongest antibacterial effects. The AgNPs exhibited broad-spectrum antibacterial activity against pathogens such as Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, and fish pathogens like Aeromonas veronii. Microwave-assisted synthesis with ethanolic extracts resulted in the highest inhibition, particularly against fish and tuberculosis-related pathogens, including Mycobacterium marinum. Nanoparticle formation was confirmed using various characterization methods, including ultraviolet-visible (UV-Vis) spectroscopy, X-ray diffraction (XRD), fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), which revealed crystalline structures. Transmission electron microscopy (TEM) analysis revealed that AgNPs varied in size, with an average diameter of less than 50 nm and all particles being smaller than 100 nm. This research demonstrates the potential of AgNPs as an effective alternative to antibiotics, offering targeted bacterial inhibition while reducing the risk of antibiotic resistance. This makes it a promising approach for treating bacterial infections in ornamental fish.
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Biomass Production of Chlorella vulgaris var. vulgaris TISTR 8261 During Cultivation in Modified Food Industry Wastewater
(2025-12-01)
Taikhao, Samart
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Phunpruch, Saranya
Industrial wastewater can serve as a low-cost nutritional source for sustainable microalgal biomass production. This study investigated the biomass of Chlorella vulgaris var. vulgaris TISTR 8261 grown in untreated wastewater collected from four food industry factories in Phra Nakhon Sri Ayutthaya Province, Thailand. Among them, wastewater from a processed food production plant (PFPP) supported the highest algal growth. Supplementation with 17.4 mM sodium acetate significantly improved algal biomass yield. Further optimization with 3.7 mM NH<inf>4</inf>Cl, 1.0 mM KH<inf>2</inf>PO<inf>4</inf>, 0.2 mM MgSO<inf>4</inf>, and a moderate concentration of trace minerals enhanced the specific growth rate and chlorophyll concentration. Scaled-up cultivation in 3.5 L culture bottles in optimized PFPP yielded a maximum biomass yield of 8.436 ± 0.378 g L−1, comparable to 6.498 ± 0.436 g L−1 in standard TAP medium. Biomass composition analysis after 15 days of cultivation revealed 42.70 ± 1.40% protein, 17.10 ± 1.60% carbohydrate, and 1.90 ± 0.10% lipid on a dry weight basis. These findings demonstrate that optimized PFPP wastewater can effectively support high-density cultivation of C. vulgaris var. vulgaris TISTR 8261, yielding nutritionally rich biomass, and offering a cost-effective and environmentally sustainable strategy for industrial-scale microalgal production.
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Physiological and Environmental Factors Influencing Hydrogen Production by Unicellular Green Alga Monoraphidium sp. KMITL-1
(2025-10-01)
Krutpan, Varanya
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Supakriangkrai, Thaninthorn
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Lohasupthawee, Pana
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Phunpruch, Saranya
With the growing global energy demand and the urgent need to reduce carbon emissions, hydrogen (H<inf>2</inf>) has emerged as a promising clean energy carrier. Among various biological H<inf>2</inf> production, green algae present a sustainable and eco-friendly alternative due to their ability to produce H<inf>2</inf> via photobiological pathways. This study aimed to investigate H<inf>2</inf> production by unicellular green alga Monoraphidium sp. KMITL-1, isolated from hydroponic water at the Plant Tissue Culture Laboratory, King Mongkut’s Institute of Technology Ladkrabang. The taxonomic identity of the strain, belonging to the genus Monoraphidium within the Selenastraceae family, was confirmed through morphological observation and molecular characterization using 23S plastid rRNA gene sequencing. Various physiological and environmental parameters influencing H<inf>2</inf> production were evaluated, including cell age, cell density, nutrient deprivation, carbon source, pH, temperature, and light intensity. A 24-hour-old culture with an OD<inf>750</inf> of 0.8 exhibited a significant increase in H<inf>2</inf> production. The optimal medium was potassium-deprived Tris-acetate-phosphate (TAP-K) supplemented with glucose at a concentration of 350 mmol C-atom L-1. The ideal environmental conditions for H<inf>₂</inf> production were pH 7.2, a temperature of 30 °C, and a light intensity of 60 μmol photons m-2 s-1. Under these optimized conditions, Monoraphidium sp. KMITL-1 achieved a maximum H<inf>2</inf> production rate of 67.976 ± 1.096 μmol H<inf>2</inf> mg Chl-1 h-1 and a cumulative H<inf>2</inf> yield of 3,190.436 ± 2.219 μmol H<inf>2</inf> mg Chl-1 after 72 h of incubation. These results highlight the potential of Monoraphidium sp. KMITL-1 for large-scale biohydrogen production and its applicability in the development of sustainable energy technologies.
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Combined Effect of Acid Salts with Clove and Cinnamon Oils on Controlling of Postharvest Decay in Carrot
(2025-09-03)
Nanasombat, Suree
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Phunpruch, Saranya
Fungal decay in carrots is a serious problem during postharvest storage. The present work aimed to study the antifungal effect of ammonium carbonate (AC) or potassium metabisulfite (PM) in combination with cinnamon or clove oils on delaying carrot decay caused by black rot mold during chill storage. In vitro trials were conducted to determine the antifungal activity of those acid salts, essential oils and their combinations against selected fungi isolated from carrots. Alternaria sp. C7D7 isolated from carrots was inhibited by AC and PM at 0.5-1.0% w/v minimum inhibitory concentration (MIC), while cinnamon and clove oils possessed strong anti-Alternaria effect with 0.025% w/v MIC. The combination of AC or PM at 0.25-0.5% w/v with cinnamon or clove oils at 0.04-0.25% w/v in potato dextrose agar produced a 100% antifungal index. Synergy testing by the checkerboard method showed that the combination of AC and cinnamon oil had partial synergistic effects against the mold. The optimum concentrations of combined AC or PM and cinnamon or clove oil were selected to formulate dipping solutions for carrot treatments. Their effect on controlling black rot decay in carrots during chill storage was investigated. The treatment with the dipping solutions consisting of combined 0.5% AC with 0.25% cinnamon oil or 1% AC alone caused a lower percentage of black rot decay (13.9-19.4% decay) in carrots compared to the others during storage at 5°C for 10 weeks. These findings show the usefulness of cinnamon oil and AC or their combination on delaying black rot decay in carrots.
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Light-Emitting Diode Illumination Enhances Biomass, Pigment, and Lipid Production in Halotolerant Cyanobacterium Aphanothece halophytica
(2025-06-01)
Thongtha, Sitthichai
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Kittiwongwattana, Chokchai
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Incharoensakdi, Aran
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Phunpruch, Saranya
Light characteristics, including spectrum and intensity, significantly impact cyanobacterial biomass production, pigment biosynthesis, and cellular metabolism, influencing the composition of various biochemical compounds. This study aimed to investigate the effects of light-emitting diode (LED) illumination on biomass, pigment, and lipid production in the unicellular halotolerant cyanobacterium Aphanothece halophytica, cultivated in a suitable natural seawater (SNSW) medium. The results revealed that LED light outperformed fluorescent light, with blue LED light, particularly at an intensity of 60 μmol photons m−2 s−1, significantly enhancing growth, pigment synthesis, and lipid accumulation. This resulted in a maximum cell density of 68.96 ± 1.52 × 106 cells mL−1, a specific growth rate of 0.302 ± 0.002 day−1, and a lipid productivity of 56.81 ± 0.75 mg L−1 day−1. White LED light produced lipids suitable for biodiesel, whereas blue, green, and red LEDs promoted the accumulation of polyunsaturated fatty acids (PUFAs), beneficial for food supplements. These findings highlight the potential of LED-based cultivation strategies for optimizing biomass and biochemical compound production in A. halophytica.
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Occurrence of Parasitic and Bacterial Pathogen in Ornamental and Wild Populations of Siamese Fighting Fish (Betta splendens) in a Region of Thailand
(2025-05-01)
Manklinniam, Piyapan
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Phunpruch, Saranya
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Worananthakij, Worakrit
The diversity of fish parasites reflects the diversity of parasites in the water source, providing insights into the effects of pathogens and essential information about parasite-host relationships. Parasitic infections are valuable indicators of the aquatic ecosystem, influenced by various factors. This study aimed to investigate the prevalence and seasonality of parasite species and pathogenic bacteria in Siamese fighting fish (Betta splendens) over 12 months, involving sixty ornamental Siamese fighting fish collected from commercial sources and 81 wild Siamese fighting fish from different natural habitats. Parasite prevalence in ornamental Siamese fighting fish (25.00%) was lower than in the wild (34.57%). The protozoa parasite Trichodina was found in both Siamese fighting fish, with the highest prevalence recorded for Trichodina sp. in wild Siamese fighting fish. The winter season exhibited the highest parasitic prevalence, with a tremendous diversity of parasites found at each location, followed by the rainy and summer seasons. This study also reported the first finding of Henneguya sp. infection in Siamese fighting fish and on the body surface. The prevalence and seasonality of parasite genera were significant in the wild compared to ornamental Siamese fighting fish. Bacterial isolation was performed on internal organs, and isolates were identified using PCR techniques. Aeromonas veronii and Mycobacterium marinum were detected in ornamental Siamese fighting fish, while A. veronii was found in wild Siamese fighting fish. These findings indicate that infections in Siamese fighting fish display seasonal variation and are impacted by their ecology. This information is fundamental for managing the biodiversity of parasites in fish and preventing parasite infections in aquaculture.

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