Decolorization of azo dyes with high salt concentration by salt-tolerant mixed cultures under anaerobic conditions

Because the lack of detailed study of biological decolorization in high salt dye wastewater, it is still difficult to evaluate the biological treatment on high-salinity dye wastewater. The experiments were carried out to study the salt-tolerant bacteria, which is useful in the treatment of high-salinity colored wastewater. Simulated wastewater containing 5-150 g/L salt （NaCI） and 50-2000 mg/L Reactive Brilliant Red K-2BP was treated with three salt-tolerant mixed cultures （CAS, TAS, DSAS）, which were under a gradually acclimated procedure. With the increase of concentrations of salt and dye, the decolorization became low. The abilities of decolorization of dyes wastewater by three mixed cultures （CAS, TAS, DSAS） were studied, CAS and DSAS mixed cultures showed more active for the treatment of high-salinity colored wastewater than TAS mixed cultures. The results suggested that there might be a simple process for the high salt wastewater treatment, which could be incorporated into conventional activated sludge plants.

Optimization of Culture Conditions and Medium Composition for the Marine Algicidal Bacterium Alteromonas sp. DH46 by Uniform Design

Harmful algal blooms(HABs) have led to extensive ecological and environmental issues and huge economic losses.Various HAB control techniques have been developed,and biological methods have been paid more attention.Algicidal bacteria is a general designation for bacteria which inhibit algal growth in a direct or indirect manner,and kill or damage the algal cells.A metabolite which is strongly toxic to the dinoflagellate Alexandrium tamarense was produced by strain DH46 of the alga-lysing bacterium Alteromonas sp.The culture conditions were optimized using a single-factor test method.Factors including carbon source,nitrogen source,temperature,initial pH value,rotational speed and salinity were studied.The results showed that the cultivation of the bacteria at 28℃ and 180 r min-1with initial pH 7 and 30 salt contcentration favored both the cell growth and the lysing effect of strain DH46.The optimal medium composition for strain DH46 was determined by means of uniform design experimentation,and the most important components influencing the cell density were tryptone,yeast extract,soluble starch,NaNO3 and MgSO4.When the following culture medium was used(tryptone 14.0g,yeast extract 1.63g,soluble starch 5.0 g,NaNO3 1.6 g,MgSO4 2.3 g in 1L),the largest bacterial dry weight(7.36 g L-1) was obtained,which was an enhancement of 107% compared to the initial medium;and the algal lysis rate was as high as 98.4% which increased nearly 10% after optimization.

CLONAL PROLIFERATION AND LONG-TERM CULTURE OF MALIGNANT LYMPHOMA CELLS UNDER SERUM-FREE CULTURE CONDITIONS

We developed a serum-free culture system that promoted the growth of B cell colonies in peripheral blood, bone marrow, lymph nodes and cerebrospinal fluid (CSF) from 7 out of 8 patients with non-Hodgkin's lymphomas of B cell type. The culture cells were pretreated with or without galactose oxi-dase (GO) prior to plating. Colony growth was best supported with BCGF. A moderate increment was observed with rIL-3, as well as rIL-1β and even to a lesser degree, by rlL-2, while B cell stimulating factor-2 (rBCSF-2) and rlL-1β did not show significant activity. rGM-CSF and rG-CSF had little effect, while rM-CSF enhanced the formation of lymphoma colonies. The cells from different patients had different requirements for Staphylococcus aureus protein A and GO pretreatment. It reflected the differences in activation and differentiation status and surface properties of lymphoma cells from different patients. The cells from CSF of one patient were successfully maintained in serum-free culture medium supplemented with 10% BCGF or 5% PHA-LCM for more than 4 months. The long-term culture cells were EBV negative, phenotypically consistent with B cells and gene rearrangements for JH, Kappa and myc. This serum-free culture system allowed extensive analysis of the growth requirements for clonogenic precursors.

Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli

Anti-lipopolysaccharide factors (ALFs) are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In this study, we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Escherichia coli maintained in shake-flasks. In particular, we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design, and two significant screened factors, (NH4)2SO4 and KH2PO4, were further optimized via the central composite design (CCD). Based on the CCD analysis, we investigated the induction start-up time, the isopropylthio-D-galactoside (IPTG) concentration, the post-induction time, and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L (NH4)2SO4 and 3.18 g/L KH2PO4, with a cell optical density of 0.8 at 600 nm before induction, an IPTG concentration of 0.5 mmol/L, a post-induction temperature of 32.7°C, and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% (without optimization) to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation, and determined that the soluble target protein constituted 10.5% of the total protein. Our identification of the economic medium composition, optimal culture conditions, and details of the fermentation process should facilitate the potential application of ALF for further research.

The Skin Whitening Effect of Co-Cultured Conditioned Medium: Involvement of Synergy between Stem Cells and Immune Cells

Many researchers have described that mesenchymal stem cells conditioned medium and immune cells conditioned medium have a clear whitening effect when they are used as cosmetic ingredients. In this study, we confirmed the whitening efficacy of various concentrations of immune cells and stem cell conditioned media. The author tried to study a conditioned medium that has a strong whitening effect even with a composition of less than 20% (the most used concentration in cosmetics). Because of the fact that the conditioned medium contains various cytokines and growth factors secreted by stem cells or immune cells, it is known to have effects such as wound healing, antioxidant, and whitening effect. Recently, stem cells have been used not only in the development of cosmetic raw materials but also in skincare procedures, and there are reports being released of cosmetics using immune cells conditioned medium. The concentration-dependent whitening effect equivalently increased as the concentration of the mono-cultured conditioned medium was obtained through the stem cells or immune cells culture. In the case of co-culture, whitening results are like the effect of positive control such as arbutin in the medium carrying only 10% of the co-cultured conditioned medium. It is possible that enhanced whitening efficiency in co-cultured conditioned medium leads to a major innovation in the global cosmetic market.

Effects of culture conditions on ligninolytic enzymes and protease production by Phanerochaete chrysosporium in air

The production of ligninolytic enzymes and protease by Phanerochaete chrysosporium was investigated under different culture conditions. Different amounts of medium were employed in free and immobilized culture, together with two kinds of medium with different C/N ratios. Little lignin peroxidase （LIP） （〈 2 U/L） was detected in free culture with nitrogen-limited medium （C/N ratio： 56/2.2, in mmol/L）, while manganese peroxidase （MnP） maximum activity was 231 and 240 U/L in 50 and 100 ml medium culture, respectively. Immobilized culture with 50 ml nitrogen-limited medium gave the highest MnP and LiP production with the maximum values of 410 and 721 U/L separately on the day 5; however, flasks containing 100 ml nitrogen-limited medium only produced less MnP with a peak value of 290 U/L. Comparatively, carbon-limited medium （C/N ratio： 28/44, in mmol/L） was adopted in culture but produced little MnP and LiE Medium type had the greatest impact on protease production. Large amount of protease was produced due to glucose limitation. Culture type and medium volume influence protease activity corporately by affecting oxygen supply. The results implied shallow immobilized culture was a possible way to gain high production of ligninolytic enzymes.

STUDIES ON CULTURE CONDITIONS OF BENTHIC DIATOMS FOR FEEDING ABALONE I.EFFECTS OF TEMPERATURE AND LIGHT INTENSITY ON GROWTH RATE

Effects of temperature and light intensity on the groWth rate of four species of benthic diatoms for feeding abalone were studied by using a novel method (in situ optical density measurement)which was simple, convenient, fast, accurate and reliable for determining biomass of benthic algae. The results showed that the favourable temperature and light intensity were as follows: for Amphora coffeaeformis, 30℃, 18 W/m2 for Cocconeis scutellum var. parv. 18℃, 20 W/m2; for Navicula Corymbosa, 18℃. 12.4 W/m2;for Navicula mollis, 24℃, 6 W/m2.

Biological characteristics of marine bacterium S-9801 strain and its culture conditions of pigment production

Strain of Flavobacterium sp.(S-9801),was screened from 207 strains of marine bacteria isolated from the Bohai Sea continental shelf and the Zhujiang Estuary,for its red pigment production.The biological characteristics of strain S-9801 and culture conditions of pigment production have been checked out in this study. The color of the bacterial colony on 2216E medium was from coccineus to rose bengal. Optimum culture conditions were sodium chloride concentration(g/dm3),10～30;pH,3～8;temperature,25～28℃;tryptone and yeast extract as nitrogen sources and glucose as carbon source. Under optimum conditions,pigment accumulation started after 12 h,reaching a maximum rate of synthesis at 36 h.

The Influence of Culture Medium Type on Cellular Phenotype of Canine Adipose Derived Stem Cells

Canine adipose derived stem cells (ASCs) hold a great promise for the therapy of osteoarthritis in veterinary medicine. Current therapy is an autologous, stromal vascular fraction. Allogeneic ASCs provide many advantages, including efficient, cost-effective treatments while eliminating a surgical procedure in a diseased animal. Cultured ASCs can be expanded and characterized, allowing selection of desirable qualities. Use of allogeneic ASCs requires selection of a culture medium that provides consistent, desirable cellular products. The supplements within a medium can greatly influence cellular phenotypes. We hypothesized that medium type influenced cellular phenotype, allowing selection of a specified cellular product for clinical applications. We evaluated ASCs derived from adipose tissue of six dogs, assessing mRNA expression of proinflammatory: interleukin-1b, cyclooxygenase-2, and anti-inflammatory mediators: tissue inhibitor metalloproteinase-2 and interleukin -1 receptor antagonist, via quantitative RT-PCR prior to, and following culture in five cell culture media: basic cell growth medium (BGM), Keratinocyte N acetyl-L-cysteine supplemented (KNAC) medium, Multipotent Adult Progenitor Cell (MAPC) medium, serum free medium (SFM) and xeno-free medium. Major histocompatability complex I (MHCI), major histocompatability complex II (MHCII), CD44 and CD90 immunophenotypes were assessed via flow cytometry analysis. Tri-lineage differentiation (bone, adipose and cartilage tissue) was utilized to verify multipotency. SFM and xeno-free culture conditions did not produce cell expansion sufficient to assess phenotype. ASCs prior to culture had wide variability in all mediator levels, while culturing in the remaining conditions resulted in more predictable expression levels of inflammatory mediators, with a decrease in all levels. Cultured ASCs retained expression of cell surface markers MHCI, CD44 and CD90, while decreasing MHCII expression levels. KNAC and MAPC medium conditions consistently produced tri-lineage differentation;BGM, SFM and xeno-free medium did not. Culture condition will influence phenotype of ASCs, and should be selected according to the intended therapeutic effect.

Effects of Culture Conditions on Growth and Docosahexaenoic Acid Production from Schizochytrium limacinum

The effects of temperature, initial pH, salinity of culture medium, and carbon and nitrogen sources on growth and doco- sahexaenoic acid (C22 : 6 n-3, DHA) production from Schizochytrium limacinum OUC88 were investigated in the present study. The results revealed that the optimal temperature, initial pH and salinity level of the medium for DHA production were 23℃, 7.0 and 18, respectively. Glucose was proved the best carbon source for the growth and DHA production from S. limacinum. Among the nitrogen sources tested, soybean cake hydrolysate, a cheap by-product, was found to be effective for the accumulation of DHA in S. lima- cinum cells. In addition, increasing the concentration of carbon sources in the medium caused a significant increase in cell biomass; however, accumulation of DHA in cells was mainly stimulated by the ratio of C/N in the medium. Under the optimal culture condi- tions, the maximum DHA yield achieved in flasks was 4.08 g L-1 after 5 d of cultivation.

New Revolutionary Technologies of Inspection of the Condition of Architectural Monuments and Buildings After Earthquakes

The existing modern traditional methods of estimation of the technical condition of buildings structure after seismic influences do not give a full picture of integrity of a construction.Besides,these methods are not suitable for difficult complexes and responsible objects since they are based on expert estimation and do not give quantitative values of parameters.The problem of creation of

Isolation Technology of Single Spore and Optimization of Conidia Culture Condition for Ustilaginoidea virens

The single spores were isolated from chlamydospores of Ustilaginoidea virens with three different maturities by PSA. The isolated single spores were cul- tured on different media at different temperatures under natural light for inducing conidia to explore the optimum isolation technique of single spore and optimum cul- ture condition of conidia. The results showed that the successful isolating rate of single spore from yellow rice false smut balls reached 90.00%. The sporulatina quantities of isolated single spores cultured on PSD and PDB media at 22 -29 ~C （variable temperature under natural light） or 28 ℃ （constant temperature under dark condition） for 12 d were up to 6.3× 107 and 1.1× 106 spore/mL, respectively. PSA was the most effective method to isolate single spores from yellow rice false smut balls of U. virens. The optimum conidia culture condition included PSD or PDB medium, 22 -29℃ or 28℃, natural light and vibration culture. Key words Ustilaginoidea virens; Single spore isolation; Conidia; Culture condition

STUDIES ON CULTURE CONDITIONS OF BENTHIC DIATOMS FOR FEEDING ABALONE Ⅱ. EFFECTS OF SALINITY pH, NITROGENOUS AND PHOSPHATE NUTRIENTS ON GROWTH RATE

Effects of salinity , pH, nitrogenous and phosphate nutrients on the growth rate of four species of benthic diatoms were studied by using the method of in situ optical density measurement . The optimal culture conditions for the four species of diatoms are as follows: for Amphora coffeaeformis , s 35 ; pH 8.5; N03 -N 1.54, NH4-N, 7.0; CO (NH2)2-N, 2.5 (mg/L); PO43--P 1.13 (mg/L) ; for Cocconeis scutellum var parva , s 30 ; pH 8.5 ; NO3-N 3.08, NH4+-N 3.5, CO(NH2)2-N 5.0 (mg/L); pO43-－P0.283 (mg/L), for Navicula corymbosa s 25; pH 8.0; N03- －N 1.54, NH4+ -N 3.5, CO(NH2)2-N 5.0(mg/L); PO43-－P0.565 (mg/L), for Navicula mollis , s 25 ; pH 8.0; NO3- － 1.54 NH4+ - N 1.75, CO(NH2)2 -N 1.25 (mg/L), P043-－P 0.141(mg/L).

Optimization of Culture Conditions for Production of Bioactive Metabolites by <i>Streptomyces</i>spp. Isolated from Soil

The current work was carried out under a screening program targeted at isolation of bioactive Streptomyces species from soil samples. A total of 54 Streptomyces species were isolated from soil samples, out of which 4 isolates were found to be promising. These isolates were identified as Streptomyces spectabilis, Streptomyces purpurascens, Streptomyces coeruleorubidus and Streptomyces lavendofoliae and their sequences have been deposited in the GenBank. The influence of culture conditions including, incubation time, incubation temperature, initial pH and different carbon and nitrogen sources on growth and bioactive compound formation was investigated. Isolate R1, identified as Streptomyces spectabilis, showed maximum bioactive metabolite production with cellobiose and peptone as the carbon and nitrogen sources, on the 5th day at pH 5 at 30℃. The optimum conditions for production by isolate R3, identified as Streptomyces purpurascens, were observed to be starch and casein as the carbon and nitrogen sources, pH 7, temperature 30℃ and an incubation period of eight days. For isolate R5, identified as Streptomyces coeruleorubidus, maximal production resulted on the sixth day at pH 6 and temperature of 35℃ with mannitol and JBM. Isolate Y8, identified as Streptomyces lavendofoliae, was found to produce high levels of bioactive metabolites in the medium supplemented with starch and peptone on the 10th day at pH 7 and at an incubation temperature of 30℃. The four strains tested here behaved differently, each one requiring specific conditions for maximum growth as well as bioactive metabolite production.

Studies on culture condition and extracellular hydrolase of psychrophilic bacteria from Arctic sea ice

Arctic sea ice in the polar region provides a cold habitat for microbial community. Arctic sea ice microorganisms are revealed to be of considerable importance in basic research and potential in biotechnological application. This paper investigated the culture condition and extraceIlular hydrolase of 14 strains of different Arctic sea ice bacteria. The results showed that optimal growth temperature of strains is 15 ℃ or 20 ℃. The optimal pH is about 8.0. They hardly grow at acid condition. 3 % NaCl is necessary for better growth. These strains have different abilities in producing amylase, protease, eellulase and lipase. Pseudoalteronomas sp. Bsi429 and Pseudoalteronomas sp. Bsi539 produced both cellulose, protease and lipase. These results provide a basis for further developing and exploiting the cold adapted marine enzyme resources.

Optimization of Streptomyces caelestis Culture Conditions and Preliminarily Determination of Its Antifungal Components

Streptomyces caelestis FAS is an actinomyces strain isolated with Ustilago scitaminea as an indicator. With GAUZE s medium as the basic medium, carbon source and nitrogen source in the medium and ventilation volume were optimized, and the antifungal substances produced by S. caelestis FAS were preliminarily determined through organic solvent extraction and ammonium sulfate precipitation. The results showed that the antifungal components produced by S. caelestis FAS were proteins, and the optimal culture conditions were using corn flour as carbon source and KNO 3 as nitrogen source and culturing in 250 ml triangular flask added with 70 ml of culture medium. The crude protein obtained under these culture conditions had the best antifungal effect with an inhibition zone diameter of 25 mm. This study will provide a scientific basis for further study on the antifungal mechanism and application of S. caelestis FAS.

Optimization of Rooting Medium for Tissue Culture Seedlings of Dendrobium officinale by Response Surface Methodology

[Objectives] To increase the survival rate of tissue culture seedlings of Dendrobium officinale,and optimize the conditions of rooting medium by the response surface methodology( RSM). [Methods]The effects of 6-BA concentration,NAA concentration,potato amount and the amount of mashed banana on the growth of seedlings were determined by single factor experiment and were analyzed by Box-Behnken design and response surface methodology. [Results]The optimal culture conditions: rooting medium 1/2 MS + 6-BA 0. 24 mg/L + mashed banana 87. 63 g/L + potato 89. 30 g/L + NAA 0. 52 mg/L + sucrose 20. 0 g/L + activated carbon 4. 0 g/L + agar 7. 0 g/L,p H 5. 8,and light intensity 2 000 Lx. [Conclusions]The model established by response surface methodology has a good predictability and could be used to optimize the conditions of tissue culture and rooting medium of D. officinale.

Effects of nutrients and light intensity on the growth and biochemical composition of a marine microalga Odontella aurita

Algal biotechnology has advanced greatly in the past three decades.Many microalgae are now cultivated to produce bioactive substances.Odontella aurita is a marine diatom industrially cultured in outdoor open ponds and used for human nutrition.For the first time,we have systematically investigated the effects of culture conditions in cylindrical glass columns and flat-plate photobioreactors,including nutrients(nitrogen,phosphorus,silicon,and sulfur),light intensity and light path,on O.aurita cell growth and biochemical composition(protein,carbohydrate,β-1,3-glucan,lipids,and ash).The optimal medium for photoautotrophic cultivation of O.aurita contained 17.65 mmol/L nitrogen,1.09 mmol/L phosphorus,0.42 mmol/L silicon,and 24.51 mmol/L sulfur,yielding a maximum biomass production of 6.1–6.8 g/L and6.7–7.8 g/L under low and high light,respectively.Scale-up experiments were conducted with flat-plate photobioreactors using different light-paths,indicating that a short light path was more suitable for biomass production of O.aurita.Analyses of biochemical composition showed that protein content decreased while carbohydrate(mainly composed ofβ-1,3-glucan)increased remarkably to about 50%of dry weight during the entire culture period.The highest lipid content(19.7%of dry weight)was obtained under 0.11 mmol/L silicon and high light conditions at harvest time.Fatty acid profiles revealed that 80%were C14,C16,and C20,while arachidonic acid and eicosapentaenoic acid(EPA)accounted for 1.6%–5.6%and 9%–20%of total fatty acids,respectively.High biomass production and characteristic biochemical composition profiles make O.aurita a promising microalga for the production of bioactive components,such as EPA andβ-1,3-glucan.