Isolation and characterization of a new Arthrospira strain

A filamentous microorganism, morphologically similar to the cyanobacterium Arthrospira, was isolated from Mangueira Lagoon in Brazil, from which Arthrospira has not previously been isolated. Random amplified polymorphic DNA (RAPD) comparison with the standard Arthrospira platensis strains LEB 52 and Paracas indicated that the organism isolated was an Arthrospira isolate, which we denominated strain LEB 18. The RAPD analysis showed conserved sequences which indicated that the three strains belonged to the same genus, and were all Arthrospira species, but there were sufficient differences between them suggesting that they were separate strains. The strain LEB 18 was cultivated in undiluted Zarrouk medium and in 60% and 20% (v/v) Zarrouk medium diluted with sterilized Mangueira Lagoon water (MLW) using illuminance rates of 32.5, 45.5 and 58.5 micromol m(-2) s(-1) according to a complete 32 factorial design with a triplicate central point. The strains LEB 52 and Paracas were cultived in the conditions central point. Our new isolate produced the highest specific growth rate (Umax = 0.22 d(-1)) in 60% Zarrouk medium diluted with MLW and illuminated with 58.5 micromol m(-2) s(-1) and the highest protein content (86.0% w/w).     

Dilution of solar radiation through "culture" lamination in photobioreactor rows facing south-north: a way to improve the efficiency of light utilization by cyanobacteria (Arthrospira platensis)

Efficient utilization of solar radiation for the photoautotrophic production of cyanobacterium biomass was achieved, using small pipes (ID = 0.01 m) arranged in rows in two photobioreactors facing south-north. A high Arthrospira yield of 47.7 g m(-2) (installation area) d(-1) was attained under outdoor conditions in a tubular undulating row photobioreactor (TURP-10r). During the summer, under a semicontinuous culture regime, the optimal biomass concentration (OBC) in TURP-5r was 6.0 g L(-1): it was 5.0 g L(-1) in TURP-10r. These OBCs made it possible to produce a biomass output rate of 2.7 +/- 0.2 g L(-1) d(-1) in the former and 2.1 +/- 0.1 g L(-1) d(-1) in the latter. When Arthrospira was grown at a preset dilution rate (0.3 d(-1)), sunrise cell density (SrCD) variations were not proportional to the drop of solar radiation. The SrCD was comparatively high at high solar radiation and decreased abruptly with decreasing solar radiation. There was a tendency to stabilize at low solar radiation. In both photobioreactors, the chlorophyll content of the Arthrospira biomass (% of the dry weight) was higher at sunrise than at sunset. A comparison of the chlorophyll biomass content in the TURPs showed no significant differences. Night biomass losses were very high (> 30% of the daylight productivity) when the culture temperature was kept constant at 31 +/- 1.0 degrees C: these losses fell to < 20% of the daylight productivity, when the night temperature of the cultures decreased according to the environmental temperature. Dilution of solar radiation was carried out using two quasi-laminated bioreactors. The rows of S-N facing bioreactors showed a very high growth yield in TURP-10r [about 2.1g (d.w.) MJ(-1)]. In TURP-10r, the high photic ratio (R(f) = 6), the high surface-to-volume ratio (S(ill)/V = 400 m(-1)) and the S-N facing of the rows (better than an E-W orientation) allowed for good results.     

Optimal temperature and photoperiod for the cultivation of Agardhiella subulata microplantlets in a bubble-column photobioreactor

The optimal temperature and illumination photoperiod requirements for the phototrophic growth of a novel microplantlet suspension culture derived from the macrophytic marine red alga Agardhiella subulata were determined. The optimal growth temperature was 24 degrees C. The effects of illumination light-dark (LD) photoperiod (hour of light:hours of darkness within a 24 h cycle) on biomass production was studied within a bubble-column photobioreactor. The 4.5 cm diameter photobioreactor was maintained at near-saturation conditions with respect to light flux (38 mciromol photons m(-2) s(-1)), nutrient medium delivery (20% nutrient replacement per day), and CO(2) delivery (0.35 mmol CO(2) L(-1) h(-1)) so that the cumulative effects of photodamage on the cell density versus time curve at photoperiods approaching continuous light could be observed. Biomass production was maximized at 16:8 LD, where biomass densities exceeding 3.6 g dry cell mass L(-1) were achieved after 60 days in culture. Biomass production was proportional to photoperiod at low fractional photoperiods (< or =10:14 LD), but high fractional photoperiods approaching continuous light (> or = 20:4 LD) shut down biomass production. Biomass production versus time profiles under resource-saturated cultivation conditions were adequately described by a cumulative photodamage growth model, which coupled reversible photodamage processes to the specific growth rate. Under light-saturated growth conditions, the rate constant for photodamage was kd = 1.17 +/- 0.28 day(-1) (+/-1.0 SE), and the rate constant for photodamage repair was kr = 5.12 +/- 0.95 day(-1) (+/-1.0 SE) at 24 degrees C.     

Influence of ammonium chloride feeding time and light intensity on the cultivation of Spirulina (Arthrospira) platensis

This study dealt with the influence of both the feeding time and light intensity on the fed-batch culture of the cyanobacterium Spirulina (Arthrospira) platensis using ammonium chloride as a nitrogen source. For this purpose, a 2(2) plus star central composite experimental design combined with response surface methodology was employed, and the maximum cell concentration (X(m)), the cell productivity (P(X)), and the yield of biomass on nitrogen (Y(X/N)) were selected as the response variables. The optimum values of X(m) (1,833 mg L(-1)) and Y(X/N) (5.9 g g(-1)) estimated by the model at light intensity of 13 klux and feeding time of 17.2 days were very close to those obtained experimentally under these conditions (X(m) = 1,771 +/- 41 mg L(-1); Y(X/N) = 5.7 +/- 0.17 g g(-1)). The cell productivity was a decreasing function of the ammonium chloride feeding time and a quadratic function of the light intensity. The protein and lipid contents of dry biomass collected at the end of cultivations were shown to decrease with increasing light intensity.     

Kinetic and growth parameters of Arthrospira (Spirulina) platensis cultivated in tubular photobioreactor under different cell circulation systems

Arthrospira platensis was cultivated in tubular photobioreactor in order to evaluate growth and biomass production at variable photosynthetic photon flux density (PPFD = 60, 120, and 240 μmol photons m(-2)s(-1)) and employing three different systems for cell circulation, specifically an airlift, a motor-driven pumping and a pressurized system. The influence of these two independents variables on the maximum cell concentration (X(m)), cell productivity (P(x)), nitrogen-to-cell conversion factor (Y(X/N) ), photosynthetic efficiency (PE), and biomass composition (total lipids and proteins), taken as responses, was evaluated by analysis of variance. The statistical analysis revealed that the best combination of responses' mean values (X(m) = 4,055 mg L(-1), P(x) = 406 mg L(-1)day(-1), Y(X/N) = 5.07 mg mg(-1), total lipids = 8.94%, total proteins = 30.3%, PE = 2.04%) was obtained at PPFD = 120 μmol photons m(-2)s(-1); therefore, this light intensity should be considered as the most well-suited for A. platensis cultivation in this photobioreactor configuration. The airlift system did not exert any significant positive statistical influence on the responses, which suggests that this traditional cell circulation system could successfully be substituted by the others tested in this work.     

Physico-chemical parameters influencing DNase activity of the cyanobacterium Spirulina platensis

The effects of temperature, Mg2+, EDTA concentration and rinsing on extra- and intra-cellular DNase activity of Spirulina platensis strain SSP-14, were investigated. The results indicate that the tested strain contains very high extra- and intracellular DNase activity, which actually hinders the transfer of foreign gene(s) to S. platensis, a cyanobacterium with multiple economic potentials. The extracellular DNase activity could easily be removed by rinsing the cells with Zarrouk medium more than once. The intracellular DNase activity could also be inhibited by (1) removal of Mg2+, (2) maintaining EDTA concentration above 1 mmol l(-1), and (3) manipulating below 0-4 degrees C, during all the incubation procedures. We suggest that, by using one or more of, or combining, all those experimental conditions, the chances of foreign DNA attempted to be introduced into S. platensis without being digested would be increased.     

Impact of ammonia concentration on Spirulina platensis growth in an airlift photobioreactor

Spirulina platensis was cultivated in a bench-scale airlift photobioreactor using synthetic wastewater (total nitrogen 412 mg L(-1), total phosphorous 90 mg L(-1), pH 9-10) with varying ammonia/total nitrogen ratios (50-100% ammonia with balance nitrate) and hydraulic residence times (15-25 d). High average biomass density (3500-3800 mg L(-1)) and productivity (5.1 g m(-2) d(-1)) were achieved when ammonia was maintained at 50% of the total nitrogen. Both high ammonia concentrations and mutual self-shading, which resulted from the high biomass density in the airlift reactor, were found to partially inhibit the growth of S. platensis. The performance of the airlift bioreactor used in this study compared favorably with other published studies. The system has good potential for treatment of high strength wastewater combined with production of algae for biofuels or other products, such as human and animal food, food supplements or pharmaceuticals.     

Batch kinetics of Pseudomonas sp. growth on benzene. Modeling of product and substrate inhibitions

Batch tests of benzene degradation were performed in liquid phase at 30 degrees C, pH 6.8 +/- 0.2, and 200 rpm in two 3-L stirred tank bioreactors, using the benzene-degrading bacterium Pseudomonas sp. NCIMB 9688. A relatively high starting biomass level (220-270 mg(X)/L) and starting benzene concentration ranging from 20 to 200 mg(S)/L were selected as conditions to investigate possible inhibition phenomena. Volumetric as well as specific rates of biomass formation and substrate consumption were calculated from experimental data of both growth and benzene degradation and used to propose and check a new overall kinetic model for cell growth simultaneously accounting for both product and substrate inhibitions. The results of the present study evidenced the occurrence of a competitive-type product inhibition due to 2-hydroxymuconic semialdehyde (K(iP)' = 0.902 mg(S)/L), which was stronger than the uncompetitive-type inhibition exerted by substrate (K(iS) = 7.69 mg(S)/L).     

Manganese peroxidase production by immobilized Phanerochaete chrysosporium BKM-F-1767 in a cell bioreactor

Manganese-dependent peroxidase (MnP) production was performed in an immobilized cell bioreactor in which Phanerochaete chrysosporium BKM-F-1767 was immobilized on polystyrene foam. The immobilized cell culture yielded significantly greater MnP activity than the conventional stationary liquid culture. Cultivation was carried out in batch mode; the effect of glucose concentration was investigated and growth kinetics parameters were found as, micromax=0.59 day(-1), Ks=0.33 g/L and Kss=14.5. Batch operation led to maximum MnP (770.82 U/L) in the culture medium containing 0.05% Tween 80, 10 g/L glucose, and 174 microM Mn2+ at 37 degrees C and pH 4.5. Enzyme productivity was obtained as 110.12 U/day/L.     

Effect of different thermal conditions on the pre-imaginal biology of Culex apicinus (Philippi, 1865) (Diptera: Culicidae).

The time of development and survival of the pre-imaginal period and the adult body size of Culex apicinus were analyzed in individuals reared from the 1st instar larva to adult emergence under laboratory and field conditions. In the laboratory, insects were exposed to three constant temperatures (15, 20, and 25 degrees C) and a photoperiod of 14:10 (L:D). In the field, temperature and photoperiod were not manipulated; during the study period water temperature ranged between 15.5 and 24.2 degrees C, and photoperiod changed from 13:11 to 14:10 (L:D). Survival to the imaginal stage at 15 degrees C (85.4%) and in the field (88.8%) was higher than that at 20 degrees C (45.8%) and 25 degrees C (8.3%). Based on femur and wing length measurements, the mosquitoes reared under field conditions (mean water temperature = 20.5 degrees C) were smaller than those laboratory-reared, even when the time of development in the field (29.5 - 36 days) was longer than that observed at 15 degrees C (24 days), 20 degrees C (15 days), and 25 degrees C (11 days). Male emergence was earlier than that of females (protandry) only in the field. Although laboratory conditions included a photoperiod and temperature range comparable with the observed values in the field, the disagreements between field and laboratory results suggest that the characteristics examined in this work could be affected by the joint variation of several factors not controlled in field conditions, such as photoperiod, temperature regime, and/or food quality of larval habitat.     

CO₂ from alcoholic fermentation for continuous cultivation of Arthrospira (Spirulina) platensis in tubular photobioreactor using urea as nitrogen source

Carbon dioxide released from alcoholic fermentation accounts for 33% of the whole CO(2) involved in the use of ethanol as fuel derived from glucose. As Arthrospira platensis can uptake this greenhouse gas, this study evaluates the use of the CO(2) released from alcoholic fermentation for the production of Arthrospira platensis. For this purpose, this cyanobacterium was cultivated in continuous process using urea as nitrogen source, either using CO(2) from alcoholic fermentation, without any treatment, or using pure CO(2) from cylinder. The experiments were carried out at 120 μmol photons m(-2) s(-1) in tubular photobioreactor at different dilution rates (0.2 ≤ D ≤ 0.8 d(-1) ). Using CO(2) from alcoholic fermentation, maximum steady-state cell concentration (2661 ± 71 mg L(-1) ) was achieved at D = 0.2 d(-1) , whereas higher dilution rate (0.6 d(-1) ) was needed to maximize cell productivity (839 mg L(-1) d(-1) ). This value was 10% lower than the one obtained with pure CO(2) , and there was no significant difference in the biomass protein content. With D = 0.8 d(-1) , it was possible to obtain 56% ± 1.5% and 50% ± 1.2% of protein in the dry biomass, using pure CO(2) and CO(2) from alcoholic fermentation, respectively. These results demonstrate that the use of such cost free CO(2) from alcoholic fermentation as carbon source, associated with low cost nitrogen source, may be a promising way to reduce costs of continuous cultivation of photosynthetic microorganisms, contributing at the same time to mitigate the greenhouse effect.     

Influence of cultivation conditions on xylose-to-xylitol bioconversion by a new isolate of Debaryomyces hansenii

About 270 yeast isolates were screened for xylitol production using xylose as the sole carbon source. The best isolate, Debaryomyces hansenii UFV-170, released 5.84 g L(-1) xylitol from 10 g L(-1) xylose after 24 h, corresponding to a yield of xylitol on consumed substrate (Y(P/S)) of 0.54 g g(-1). This strain was cultivated batch-wise at variable starting concentrations of xylose (S(o)) and biomass (X(o)) and agitation intensity, in order to improve xylitol production and to evaluate, through simple carbon balances, the influence of these conditions on xylose metabolism. Under the best microaerobic conditions (S(o) = 53 g L(-1), X(o) = 1.4 g L(-1), 200 rpm), xylitol production reached 37.0 g L(-1), corresponding to xylitol volumetric productivity of 1.0 g L(-1)h(-1), specific productivity of 0.22 g g(-1)h(-1) and Y(P/S) = 0.76 g g(-1). Almost 83% of xylose was consumed for xylitol production, the rest being consumed for growth, while respiration was negligible. The new isolate appeared to be a promising alternative for industrial xylitol bioproduction.     

Effects of light intensity and dilution rate on the semicontinuous cultivation of Arthrospira (Spirulina) platensis. A kinetic Monod-type approach

Semicontinuous cultures were carried out at different dilution rates (D) and light intensities (I) to determine the maximum productivity of Arthrospira platensis cultivated in helicoidal photobioreactor up to the achievement of pseudo-steady-state conditions. At I=108 μmol photons m(-2) s(-1), the semicontinuous regime ensured the highest values of maximum cell concentration (X(m)=5772±113 mg L(-1)) and productivity (P(XS)=1319±25 mg L(-1) d(-1)) at the lowest (D=0.1 day(-1)) and the highest (D=0.3 day(-1)) dilution rates, respectively. A kinetic model derived from that of Monod was proposed to determine the relationship between the product of light intensity to dilution rate (ID) and the cell productivity, which were shown to exert a combined influence on this parameter. This result put into evidence that pseudo-steady-state conditions could be modified according to circumstances, conveniently varying one or other of the two independent variables.     

Bioeffects of selenite on the growth of Spirulina platensis and its biotransformation

The bioeffects of selenium on the growth of Spirulina platensis and the selenium distribution were investigated. S. platensis was batch cultured in Zarrouk medium containing increasing concentrations of sodium selenite. The biotransformation characteristic of selenium was analysed by the determination of the detailed selenium distribution forms. At 35 degrees C, 315.2 microEm(-2) x s(-1), sodium selenite concentrations below 400 mg x l(-1) were found to stimulate algal growth, especially in the range of 0.5-40 mg x l(-1). However, above 500 mg x l(-1) sodium selenite was toxic to this alga with the toxicity being related to the sulfite level in the medium. S. platensis was found to resist higher selenite by reducing toxic Se(IV) to nonsoluble Se(0). Selenium was accumulated efficiently in S. platensis during cultivation with accumulated selenium increasing with selenite concentration in the medium. It was demonstrated that inorganic selenite could be transformed into organic forms through binding with protein, lipids and polysaccharides and other cell components. The organic selenium accounted for 85.1% of the total accumulated selenium and was comprised of 25.2% water-soluble protein-bound, 10.6% lipids-bound and 2.1% polysaccharides-bound selenium. Among the organic fractions lipid possessed the strongest ability to accumulate Se (6.47 mg x kg(-1)). The 14.9% inorganic selenium in S. platensis was composed of Se(IV) (13.7%) and Se(VI) (1.2%).     

钝顶螺旋藻在不同光照条件下的放氧特性

钝顶螺旋藻在持续照光和中等频率 (0.01~20 Hz) 的光/暗交替照光下的放氧特性对光生物反应器的设计和操作具有重要意义。构建了一套可实现光/暗交替的光生物反应器系统对此进行研究,结果显示：根据与放氧速率的关系,可以将光强分为4个区：光限制区 (0~335 μmol/(m2·s)),过渡区 (335~875 μmol/(m2·s)),光饱和区 (875~2 775 μmol/(m2·s)) 以及光抑制区 (2 775 μmol/(m2·s)以上)。提高光/暗频率能否提高微藻光合速率取决于所采用的光强和     

A two-plane tubular photobioreactor for outdoor culture of Spirulina

A photobioreactor in the form of a 245-m-long loop made of plexiglass tubes having an inner diameter of 2.6 cm was designed and constructed for outdoor culture of Spirulina. The loop was arranged in two planes, with 15 8-m-long tubes in each plane. In the upper plane, the tubes were placed in the vacant space between the ones of the lower plane. The culture recycle was performed either with two airlifts, one per plane, or with two peristaltic pumps. The power required for water recycle in the tubular photobioreactor, with a Reynolds number of 4000, was 3.93 x 10(-2) W m(-2). The photobioreactor contained 145 L of culture and covered an overall area of 7.8 m(2). The photobioreactor operation was computer controlled. Viscosity measurements performed on Spirulina cultures having different biomass concentrations showed non-Newtonian behavior displaying decreasing viscosity with an increasing shear rate. The performance of the two-plane photobioreactor was tested under the climatic conditions of central Italy (latitude 43.8 degrees N, longitude 11.3 degrees E). A biomass concentration of 3.5 g L(-1) was found to be adequate for outdoor culture of Spirulina. With a biomass concentration of 6.3 g L(-1), the biomass output rate significantly decreased. The net biomass output rate reached a mean value of 27.8 g m(-2) d(-1) in July; this corresponded to a net photosynthetic efficiency of 6.6% (based on visible irradiance). (c) 1993 John Wiley & Sons, Inc.     

Production of pullulan by a fed-batch fermentation

Summary In pullulan production from sucrose byAureobasidium pullulans, a sugar concentration higher than 5% (w/v) inhibited cell growth and the production of exopolysaccharide. By a fed-batch fermentation, the inhibitory effects of the high sugar concentration were overcome and 58.0 g/1 of exopolysaccharide were obtained from 10% sucrose.Abbreviations m, n relationship parameters for the growth and non-growth associated product formation - X, Xmax biomass and maximum biomass concentration (g cell/1) - P product concentration (g exopolysaccharide/1) - specific growth rate of cell (hr^–1)     

Influence of pH, temperature, and urea molar flowrate on Arthrospira platensis fed-batch cultivation: a kinetic and thermodynamic approach

Arthrospira platensis was cultivated photoautotrophically at 6.0 klux light intensity in 5.0-L open tanks, using a mineral medium containing urea as nitrogen source. Fed-batch experiments were performed at constant flowrate. A central composite factorial design combined to response surface methodology (RSM) was utilized to determine the relationship between the selected response variables (cell concentration after 10 days, X(m), cell productivity, P(X), and nitrogen-to-cell conversion factor, Y(X/N)) and codified values of the independent variables (pH, temperature, T, and urea flowrate, K). By applying the quadratic regression analysis, the equations describing the behaviors of these responses as simultaneous functions of the selected independent variables were determined, and the conditions for X(m) and P(X) optimization were estimated (pH 9.5, T = 29 degrees C, and K = 0.551 mM/day). The experimental data obtained under these conditions (X(m) = 749 mg/L; P(X) = 69.9 mg/L.day) were very close to the estimated ones (X(m) = 721 mg/L; P(X) = 67.1 mg/L.day). Additional cultivations were carried out under the above best conditions of pH control and urea flowrate at variable temperature. Consistently with the results of RSM, the best growth temperature was 29 degrees C. The maximum specific growth rates at different temperatures were used to estimate the thermodynamic parameters of growth (DeltaH* = 59.3 kJ/mol; DeltaS* = -0.147 kJ/mol.K; DeltaG* = 103 kJ/mol) and its thermal inactivation (DeltaH(D) (o) = 72.0 kJ/mol; DeltaS(D) (o) = 0.144 kJ/mol.K; DeltaG(D) (o) = 29.1 kJ/mol).