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Bacillus genus are Gram-positive, rod shaped, spore-forming, aerobic or facultative anaerobic bacteria, which have the ability to produce a wide variety of enzymes, antimicrobial compounds, vitamins, and carotenoids. Nowadays, Bacillus species have been increasingly proposed for use as probiotics or feed additives as B. amyloliquefaciens, B. licheniformis, B. subtilis, and B. pumilus because of their advantages such as heat-stability of the spores, storage capacity at ambient temperature, and beneficial properties for health. Since microbes are used for customers and livestock, it is critical to substantiate not only the health benefits and efficacy of unique strains but also their safety for the hosts. This study aims to evaluate the in vitro safety and some beneficial properties of Bacillus strains to select potential strains for use in humans or animals. A total of 76 Bacillus strains belonged to the B. amyloliquefaciens and B. subtilis groups were preliminary evaluated for their safety via hemolytic activity. Six non-hemolytic strains were identified and their antibiotic susceptibility, cytotoxicity on the growth of HT29 and Vero cells, extracellular enzyme production, antimicrobial activity, and identification based on 16S rRNA and rpoB gene sequences were studied. Two strains, B. subtilis VTCC 10963 and B. subtilis VTCC 11039, should be considered safe. In addition, these two strains exhibited good extracellular enzyme production (amylase, cellulase, and protease) and strong antibacterial activity against Listeria monocytogenes. Our results support the use of B. subtilis VTCC 10963 and B. subtilis VTCC 11039 for the development of probiotic products and feed additives in the future.

Pelodiscus sinensis is a widespread species of soft-shelled turtle that is commercially cultured in freshwater and prominently exploited as speciality food in Vietnam. During the cultivation, there are distinctly recurrent epidemic diseases causing economically vast damage for farmers. In this research, we carried out the isolation of microorganisms from typical ulcers “ba dau” presenting on soft-shelled turtle taken in Nam Dinh farm. The samples were inoculated on common media used for bacteria, actinomyces, fungus and yeast. Only bacteria grew on typical culture medium. Fifteen bacterial strains were isolated and identified by 16S rRNA sequence analysis. They were belonged to genera such as Acidovorax (n=1), Acinetobacter (n=1), Aeromonas (n=8), Citrobacter (n=1), Chryseobacterium (n=1), Moraxella (n=1), Paludibacterium (n=1) and Parabacteroides (n=1). In which, eight strains Aeromonas spp. were selected for further research on virulence characterization. PCR results showed that these strains possessed at least 4/7 virulence genes including aer (75%), alt (37.5%), ast (50%), ela (87.5%), exu ( 100%), hlyA (37.5%) and ser (100%). With additional tests, we confirmed the virulent expression by haemolysin (100%), protease (75%), DNAase (75%), elastase (25%) and cytotoxicity (62.5%). These results are scientific basis to establish and develop practical methods for controlling and preventing "ba dau" disease in the soft-shelled turtle in the future.

A phytochemical investigation of methanol extract from leaves of Pachyrhizus erosus (L.) Urban, a leguminous shrub distributed in Vietnam and other tropical and subtropical countries led to the isolation of a new prenylated chalcone, erosusone (1) and a new megastigmane glycoside epimer, 3-episedumoside F1 (9), together with thirteen known compounds including flavonoids (2-6), a 3-benzoxepine lactone (7), a pyridine-4,5-diol derivative (8), megastigmanes and megastigmane glycosides (10-15). Their structures were elucidated by means of high resolution-electrospray ionization (HR-ESI)-MS, one dimensional (1D) and two-dimensional NMR (2D-NMR) spectroscopy as well as comparison with the data reported in the literature. The cytotoxic effects on LU-1 (lung carcinoma), HepG2 (hepatocellular carcinoma), and MCF7 (breast carcinoma) cell lines were assessed. Prenylated chalcones 1-2 and isoflavone 3 exhibited cytotoxicity against all tested cell lines with IC50 values ranging from 22.04 to 45.03 µM.

A phytochemical investigation of methanol extract from leaves of Pachyrhizus erosus (L.) Urban, a leguminous shrub distributed in Vietnam and other tropical and subtropical countries led to the isolation of a new prenylated chalcone, erosusone (1) and a new megastigmane glycoside epimer, 3-episedumoside F

Green macroalgae have gained attention as promising renewable sources for biorefining. Despite the wide potential availability of green macroalgae, their utilization has been limited to ethanol production, hindering their further application. In this study, we report that a bacterium, Halomonas sp. strain BL6 (isolated from a mangrove forest in Bach Long, Nam Dinh Province, Vietnam), produces pyruvate from a saccharified solution of the green seaweed Ulva reticulata and secretes it into the medium. Pyruvate, an important α-oxocarboxylic acid, plays a central role in energy and carbon metabolism in living organisms and is used mainly for the synthesis of various chemicals and polymers or as an ingredient or additive in food, cosmetics, and pharmaceuticals. To investigate the possibility of using U. reticulata from the seashore of Vietnam as biomass feedstock, the chemical composition and saccharification yield of this seaweed were studied. Dry biomass of U. reticulata was found to contain 65.5% carbohydrate, 10.3% protein, 1.8% lipid, and 10.6% ash. Reducing sugar content reached 608.79 mg g−1 of biomass after pretreatment with diluted acid and 24 h of incubation with 50 IU g−1 Viscozyme L. The resulting sugars were fermented by Halomonas sp. strain BL6 to produce pyruvate, and the maximal pyruvate concentration reached 55.23 g L−1 after 72 h of cultivation. This study is the first to report the production of valuable compounds other than bioethanol products, such as pyruvate, from U. reticulata hydrolysate by a Halomonas strain.

Pyruvic acid (pyruvate) is a central intermediate in carbon and energy metabolism in all organisms. It is widely used in the industrial biosynthesis of high - value compounds and food additives. Biotechnological pyruvate production has attracted attention as a potential alternative method of pyruvate synthesis. From 27 soil and mud samples collected from mangrove forests of Cam Ranh, Khanh Hoa province, we isolated 10 bacterial strains belonging to Halomonas genus. Among them, 9 strains were able to synthesis and secreted pyruvate in the culture medium. The maximal pyruvate production was in MC8 strain with value of 0.110 ± 0.015 g/L. MC8 strain was Gram-negative, short rod-shaped, 1,56 ± 0,07 in width, 5,09 ± 0,38 µm in length, non-flagella and nonmotile. MC8 strain was positive for oxidase and catalase activities and reduction of nitrate to nitrite and nitrogen. Cells were able to growth at salt concentrations of from 0.5 to 20%, temperature ranging from 20 to 45ºC and pH ranging from 5 to 12. The major fatty acids of MC8 biomass were C16:0; C18:1w7c và C12:0 3OH. Phylogenetic analyses based on 16S rRNA gene sequencing showed that MC8 strain possessed the highest similarity of 98.5% to the strain Halomonas flava. Based on morphological, biochemical characteristics and 16S rRNA gene sequencing, MC8 strain was identified as H. flava. This is the first study of Halophilic bacteria which is capable of pyruvate synthesis in Vietnam. Thus, these obtained results provided important insights into the production of pyruvate using Halomonas strains.

Pyruvic acid (pyruvate) the most important α-oxocarboxylic acid, plays a central role in energy metabo-lism in living organisms. It is used mainly as a starting material in the biosynthesis of pharmaceuticals as well as employed in the production of crop protection agents, polymers, cosmetics and food additives. Four Gram-negative, moderately halophilic, pyruvate and poly (3-hydroxybutyrate)- producing strains were isolated from soil samples collected from mangrove forest at Giao Thuy, Nam Dinh province. Preliminary screening results showed that among fours strain, the ND7 strain was able to produce the highest bioplastic-poly (3-hydroxybutyrate) (PHB) and extracellular secretion of pyruvate, respectively, reaching 50.23% dry weight and 6.87 g/L after 60 hours of cultivation. When cultured in high alkaliphilic medium (pH 9.4), the pyruvate secretion increased significantly (21.02 g/L after 60 hours of cultivation). Based on the morphological, bio-chemical characteristics and nucleotide sequence analysis of partial 16S rRNA gene, the ND7 strain was iden-tified as Halomonas maura. The results allow us to conclude that ND7 strain is a potential target for the ex-ploitation of pyruvate in a larger scale.

The green alga Chlamydomonas reinhardtii is a model organism for studying mechanism of the body in response to changes in environmental conditions, photosynthesis as well as chloroplast biogenesis and can be modified to produce high value proteins. In this study, we present results on the selection of cultural conditions for the growth of cell wall - deficient mutant C. reinhardtii (503 strain- control) and recombinant 503-2 strain expressing VP28 protein of White Spot Syndrome Virus (WSSV) in 250 ml flasks. Suitable conditions for the growth of both control and recombinant strains were TAP medium, temperature of 25 C, pH = 7, light intensity of 2 klux, C source - Trisbase, N source - nitrate and salinity of 50 - 100 mM. In these conditions, maximum cell density of the control and recombinant strains was 3.20 ± 0.08 x 106 and 2.96 ± 0.17 x 106 cells/ mL after 3 and 7 days of cultivation, respectively. The presence of vp28 gene in the recombinant microalgal strain was stably maintained at all levels of culture. The results of this paper will be a scientific basis for culturing the recombinant C. reinhardtii to obtain algal biomass which had high expression of surface VP28 protein of WSSV’s envelope for edible vaccine production vaccine by feeding in order to prevent white spot disease of shrimp.

Soybean (Glycine max (L) Merr.) is one of the major crops grown worldwide for human food and animal feed. Nanotechnology is one of the most potential tools in modern agriculture to improve crop productivity. In this paper, we investigated the effect of single and mixture cobalt nanoparticles on the change of photosynthesis - related parameters and antioxidant enzyme activity of soybean Glycine max (L) Merr “DT26” at different growth stages under pilot scale. The results showed that all form of cobalt nanoparticles treatment enhanced the photosynthesis of soybean by increasing the content of chlorophyll a, Pn and the ratio of Fv/Fm. These values tended to increase and reached the maximum value at fifth trifoliate stage and then decreased at the beginning bloom stage. The activity of antioxidant enzymes such as superoxide dismutase and peroxidase (when seeds were treated with USA single colbalt nanoparticles at concentration of 16.67 mg/kg) and catalase (with USA single colbalt nanoparticles at concentration of 0.17 mg/kg) were increased in 2.11, 1.85 and 1.60 times respectively compared to the control group (without the treatment of colbalt nanoparticles) while ascorbate peroxidase was reached highest value (increased in 2.73 times) under the condition of Vietnam single colbalt nanoparticles at the dose of 0.17 mg/kg seeds. This suggests that cobalt nanoparticles treatment has generated oxidative stresses and soybean DT26 has the self-protection mechanism by increasing the activity of antioxidant enzymes in order to reduce the amount of H2O2 and O2 which accumulates in the leaf cells.

Phototrophic dinoflagellates of the genus Symbiodinium (= zooxanthellae) are well known as symbionts of corals. They play an important role in maintaining the healthy as well as establishment of new reefs. In addition, products such as toxins or pigments from these symbiotic microalgae are potentially important for applications in the biomedical sciences, especially in new drug development. In this study, we isolated successfully a symbiotic microalga from soft coral Sinularia sp. which was collected in Cu Lao Cham coastal waters of Quang Nam province. Based on morphological characteristics and 18S rRNA gene sequences, isolated SHM8 strain was identified as Symbiodinium goreaui. This symbiont was cultured in different media such as ASP-A8, F2, ES, IMK, K, and L1. L1 culture medium supplemented with taurine concentration of 4.88 mM led to the cell density of SHM8 strain increased in 50% compared to control on day 2 in cultivation; amino acid mixture enhanced cell density by 31.7% on day 5 in cultivation. Supplementation of carbon source (bicarbonate salt of 16.8 g/L) and nitrogen source (nitrate salt of 0.055 g/L) enhanced the cell density by 40% and 27%, respectively on day 6 in culture. Obtained results in this study proved that this microalga can live independently without host. Successful isolation and cultivation of Symbiodinium microalga are one of the necessary conditions for promising researches on the natural bioactive substances from this microalgal biomass in the future in Vietnam. Citation: Dang Diem Hong, Pham Van Nhat, Hoang Thi Huong Quynh, Luu Thi Tam, Ngo Thi Hoai Thu, Nguyen Cam Ha, Hoang Thi Lan Anh, Nguyen Hoai Nam, Nguyen Thi Minh Hang, Chau Van Minh, 2017. Study on biocharacteristics of Symbiodinium sp. isolated from soft coral in coastal water of Quang Nam, south central Vietnam. Tap chi Sinh hoc, 39(3): 367-375. DOI: 10.15625/0866-7160/v39n3.10112. *Corresponding author: ddhong60vn@yahoo.com Received 19 June 2017, accepted 20 August 2017

Halophilic bacteria are receiving increasing attention for industrial chemical production processes due to their unique properties. Herein, an alkaliphilic and halophilic bacterium was isolated from a commercial Spirulina culture at Nghe An province in Vietnam and found to secrete pyruvate. Pyruvate is widely used as a starting material in the industrial biosynthesis of pharmaceuticals, and is employed for production of crop protection agents, polymers, cosmetics, and food additives. Phenotypic and chemotaxonomic characterization, and the 16S rRNA gene sequence homology with Halomonas hydrothermalis strain DSM 15,725 (99.2%) predicted that the strain belongs to the Halomonas genus, thus we named this strain as H. hydrothermalis strain C22. We investigated the biocharacteristics and capacity of strain C22 and determined the draft genome sequence comprising 3,934,166 bp with a G + C content of 60.2% encoding 3,668 proteins, 58 tRNAs, 9 rRNAs, and 1 tmRNA. Maximal pyruvate secretion reached 51.1 g/l after 84 h of cultivation. The results will facilitate future studies on the genetic and metabolic diversity of halophilic bacteria and expand our understanding of important bioprocesses in this microorganism.

Nowadays, nanotechnology is one of the most potential tools in the modern agriculture to improve productivity, adaptation to climate change and sustainable development. In this study, the effect of nanoscale zerovalent cobalt (NZVC) on the growth, photosynthetic parameters expressed by Fo (initial fluorescence), Fm (maximal fluorescence), Fv/Fm (maximal photochemical efficiency), ΦPSII (effective quantum yield of photosystem II), ETR (photosynthetic electron transport rate), Pn (net photosynthetic rate) and content of chlorophyll a and b at different stages in soybean leaves were investigated. The NZVC made from Vietnam and USA with different concentrations (0, 0.17 and 16.7 mg/kg seed) were used. It was found that when soybean seeds were treated with NZVC, the growth indices (such as the plant height, dry weights of stems and leaves and leaf area), photosynthetic parameters and chlorophyll a and b content were increased obviously compared with that of the control. These values had tended to gradually increase and reach maximum at fifth (V5) trifoliate stage, whereas they were significantly decreased at begin at flowering (R1) stage. The real productivity of the experimental formulas exceeded the control (5%), while the quality of grain was significant different among the plots. The zerovalent cobalt nanoparticle from Vietnam exhibited similar efficiency compared to those that came from USA. Our results indicated that photosynthesis was positively affected by NZVC, leading to the improved growth of soybean and therefore to improved productivity as a whole.

Schizochytrium mangrovei PQ6 contains many important polyunsaturated fatty acids, such as docosahexaenoic acid (DHA, C22: 6w-3), eicosahexaenoic acid (EPA, C20: 5w-3) and docosapentaenoic acid (DPA, C22: 5w-6). These fatty acids are essential for survival and growth of many marine fish larvae. In this paper, fresh biomass of S. mangrovei PQ6 was used for culturing rotifer Brachionus plicatilis. In the first formula (L1), rotifers were fed on baker’s yeast; in second formula (L2), rotifers were fed on mixed two microalgae, Nannochloropsis oculata and Chaetoceros gracilis; in the third formula (L3), rotifers were fed on S. mangrovei PQ6 biomass. The results indicated that S. mangrovei PQ6 biomass can replace baker’s yeast and autotrophic microalgae in feeding rotifers. Total lipid, total fatty acid and polyunsaturated fatty acid contents of rotifers fed on formula 3 (S. mangrovei PQ6) were higher than those within others. High polyunsaturated fatty acid (C ≥ 20) in omega-3 and 6 groups; DHA and DPA contents of rotifers fed on S. mangrovei PQ6 were the highest, accounting for 39.81% of total fatty acid; 41.95% and 8.24% polyunsaturated fatty acids, corresponding to 20.52; 12.15 and 2.4 mg/g of dried weight, respectively.The survival rate of black sleeper’s larvae was highest when they were fed on rotifers grown on L3 formula (51.20 ± 0.89%), followed by L2 formula (48.70 ± 2.67%) and the lowest with L1 formula (43.44 ± 1.54%) (P < 0.05). However, among three formulas, no significant difference was found in the growth of black sleeper’s larvae (P > 0.05). These results suggest that biomass of S. mangrovei PQ6 can replace traditional feed like autotrophic microalgae or baker’s yeast for biomass culture of the Brachionus plicatilis rotifer for rearing black sleeper’s larvae to enhance seed quality to meet demands of market.

Seaweed is rich source of natural bioactive compounds that could be exploited as functional ingredient for cosmetic applications. The aim of this study was to evaluate the biochemical compositions and bioactivities of 10 seaweeds collected from coastal waters of Vietnam. The present study also prepared and evaluated cream mask from mixture of seaweeds extracted with water. The results showed that

A moderately halophilic bacteria designed strain D34 was isolated from the culture medium of green microalga Dunaliella tertiolecta. The isolate was Gram-negative, aerobic, rod-shaped, approximately 0.45–0.60 mm wide and 1.25–5.10 mm long, occuring singly, non-motile, and flagellum- less. Colonies on solid media are cream, circular, and smooth. This strain was able to produce exopolysaccharide, poly hydroxybutyrate, oxidase and catalase positive. Growth occurred in a temparature range of 20–40°C, a salts concentration of 0.1–25% (w/v), and pH range 6–12. The major fatty acids were C16:0 (35.59%), C16:1w-7 (20.54%), C18: 1w-7 (30.14%), and C12:0 (10.03% of total fatty acids). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain D34 belonged to the genus Halomonas. The highest levels of 16S rRNA gene sequence similarity were found between the strain D34 and H. aquamarina (sequence similarity 98.6 %).Pyruvate, a central intermediate in metabolism processes in all organisms, is widely used for the synthesis of various chemicals and polymers as well as ingredient or additive in food, cosmetics, and pharmaceuticals. In this study, pyruvate production by strain D34 following changes in culture medium, glucose and nitrate concentrations and culture temperature were also studied. In 84 hours of batch- cultivation, pyruvate production by wild-type Halomonas sp. D34 reached 37.24 g/L at 37°C with 20% glucose and 30 g/L sodium nitrate adding to SOT medium. These data provided evidences for pyruvate production using novel wild-type strains.

Marine microalga Isochrysis sp. was isolated from Hai Phong area of Vietnam coastal zone. Their pure cultures were raised in the laboratory for culture optimization, molecular identification and biochemical analysis. Based on the morphological studies the newly isolated species was identified as Isochrysis galbana Parke and was confirmed by 18S rRNA gene analysis. The nucleotide sequence of partial 18S rRNA gene was submitted to the GENEBANK with accession number FJ536744. Optimum conditions for growth of the species was f/2 medium at temperature 25 ± 1°C, salinity 30 ± 1 psu, pH 7.0 and light intensity of 100 μmol m−2 s−1. Under these optimum conditions, the content of protein, carbohydrate and total lipid were 27.981 ± 0.892%, 25.263 ± 0.789% and 9.780 ± 0.120% of dry cell weight, respectively (equivalent 38.768 ± 0.892%, 35.000 ± 0.789% and 13.550 ± 0.120% of ash free dry weight). I. galbana Parke, strain HP has the highest content of docosahexanenoic acid (DHA), up to 14.7% of total fatty acid and maximal polyunsaturated fatty acids (PUFAs) values at the early stationary phase in f/2 medium. The n-3 and n-6 PUFA ratio (n-3/n-6) of strain HP was from 4.50 to 5.69 at different phases of growth curve. These results suggest that biomass of this strain can be used as live feed in aquaculture.

The heterotrophic marine microalga, Schizochytrium mangrovei PQ6, synthesizes large amounts of polyunsaturated fatty acids (PUFAs) with possible nutritional applications. We characterized the transcriptome of S. mangrovei PQ6, focusing on lipid metabolism pathways throughout growth. Cell growth, total lipid, and docosahexaenoic acid (DHA, 22:6n-3) contents of S. mangrovei PQ6 in 500 ml batch cultures rapidly increased on day 1 in cultivation and reached their maximum levels on day 5. Maximum lipid accumulation in 500 ml batch cultures occurred on day 5, with total lipid and DHA contents reaching 33.2 ± 1.25% of dry cell weight (DCW) and 136 mg/g DCW, respectively. 11,025 unigenes, 28,617 unigenes and 18,480 unigenes from the transcriptomes of samples collected on day 1, 3, and 5 in cultivation were identified, respectively. These unigenes of the three samples were further assembled into 30,782 unigenes with an average size of 673 bp and N50 of 950 bp, and a total of 9,980 unigenes were annotated in public protein databases. 93 unigenes involved in lipid metabolism in which expression patterns corresponded with total lipid and DHA accumulation patterns were identified. The possible roles of PUFAs pathways, such as those mediated by fatty acid synthase, polyketide synthase, and desaturase/elongase, co-exist in S. mangrovei PQ6.

Schizochytrium sp. PQ6, a heterotrophic microalga isolated from Phu Quoc (PQ) Island in the Kien Giang province of Vietnam, contains a high amount of docosahexaenoic acid (DHA, C22:6n-3). In this study, the culture conditions are developed to maximize biomass and DHA production. Nucleotide sequence analysis of partial 18S rRNA gene from genomic DNA showed that PQ6 has a phylogenetic relationship close to Schizochytrium mangrovei Raghu-Kumar. The highest growth rate and DHA accumulation of this strain were obtained in 6.0% glucose, 1.0% yeast extract, 50% artificial seawater (ASW), and pH 7 at 28°C. In addition, carbon and nitrogen sources could be replaced by glycerol, ammonium acetate, sodium nitrate, or fertilizer N-P-K. Total lipid content reached 38.67% of dry cell weight (DCW), in which DHA and eicosapentaenoic acid (EPA, C20:5n-3) contents accounted for 43.58% and 0.75% of the total fatty acid (TFA), respectively. In 5 and 10 L fermenters, the cell density, DCW, total lipid content, and maximum DHA yield were 46.50 × 10(6) cells · mL(-1) , 23.7 g · L(-1) , 38.56% of DCW, and 8.71 g · L(-1) (in 5 L fermenter), respectively, and 49.71 × 10(6) cells · mL(-1) , 25.34 g · L(-1) , 46.23% of DCW, and 11.55 g · L(-1) (in 10 L fermenter), respectively. Biomass of PQ6 strain possessed high contents of Na, I, and Fe (167.185, 278.3, and 43.69 mg · kg(-1) DCW, respectively). These results serve as a foundation for the efficient production of PQ6 biomass that can be used as a food supplement for humans and aquaculture in the future.

A growth promotion effect of steelmaking slag on Spirulina platensis M135 was investigated. The growth promotion effect was obtained that was 1.27 times greater than that obtained by the control by adding 500 mg L−1 of steelmaking slag and culturing for 60 days. The lipid content decreased in a concentration-dependent manner with steelmaking slag, whereas the carbohydrate content remained constant. The protein content of S. platensis M135 increased in a concentration-dependent manner with steelmaking slag when cultured at day 45. The superoxide dismutase activity of S. platensis M135 exhibited a decreasing trend in a time-dependent manner and an increasing trend in the control. The superoxide dismutase activity was lower than that of the control at day 1 but was higher at day 30. No genetic damage was observed up to 500 mg L−1 of steelmaking slag at 30 days of culture. Recovery from genetic damage was observed at 1,000 mg L−1 of steelmaking slag but not at higher concentrations.