Ferric chloride flocculation plus carbon adsorption allows to reuse spent culture medium of Arthrospira platensis

Arthrospira platensis cultivation produces a saline spent medium that must be treated to allow its reuse, thus saving water and avoiding environmental pollution. This study evaluates the association of flocculation followed by adsorption to treat the spent medium by applying different concentrations of granular activated carbon (GAC) and ferric chloride (F), and using different residence times (T). The simultaneous optimization of the independent variables GAC, F, and T was performed using both a 2³ central composite design and a response surface methodology. The cells cultivated in the medium obtained after the optimal conditions of treatment (GAC = 54.2 g L^?1, F = 10.0 mg L^?1, and T = 30.8 min) provided the highest maximum cell concentration, X_m = 3140 ± 77 mg L^?1 in 0.5 L Erlenmeyer flasks with the highest protein biomass content (44.9%). The treated medium in such conditions was also used in a 3.5 L tubular photobioreactor (PBR), reaching X_m = 4033 ± 110 mg L^?1 and biomass with high contents of both protein (47.3 ± 2.6%) and chlorophyll (9.7 ± 0.3 mg g dry cell^?1). Therefore, this study can contribute to diminishing costs of A. platensis production by reusing its culture medium and improving its biomass quality in PBRs.     

Simultaneous use of sodium nitrate and urea as nitrogen sources improves biomass composition of Arthrospira platensis cultivated in a tubular photobioreactor

Arthrospira platensis is widely cultivated in open ponds for industrial purposes. However, high‐protein A. platensis biomass produced in photobioreactors (PBRs) is recommended for pharmaceutical and cosmetic formulations. A. platensis was cultivated in a 3.5 L tubular airlift PBR using both sodium nitrate and urea as nitrogen sources. Sodium nitrate was added from the start of the cultivation using a batch process. Urea was supplied daily at exponentially increasing feeding rate using a fed‐batch process. The simultaneous optimization of the independent variables, namely, total quantity of sodium nitrate (m_T1) and total quantity of urea (m_T2), led to an optimal condition of m_T1 = 15.0 mmol/L and m_T2 = 7.5 mmol/L. Maximum biomass concentration (5183 ± 94 mg/L) corresponding to the highest biomass productivity (683 ± 13 mg/L/day) was obtained under such condition. The addition protocol of both nitrogen sources resulted in high productivities of protein (6.2 ± 0.4 mg/L/day) as well as chlorophyll‐a (372.2 ± 7.7 mg/L/day). Such innovative process could be applied in the large‐scale production of A. platensis using tubular PBR for novel applications.     

Photoautotrophic Production of Lipids by Some <Emphasis Type="Italic">Chlorella</Emphasis> Strains

The microalgae Chlorella protothecoides UTEX 25, Chlorella sp. TISTR 8991, and Chlorella sp. TISTR 8990 were compared for use in the production of biomass and lipids under photoautotrophic conditions. Chlorella sp. TISTR 8990 was shown to be potentially suitable for lipid production at 30°C in a culture medium that contained only inorganic salts. For Chlorella sp. TISTR 8990 in optimal conditions in a stirred tank photobioreactor, the lipid productivity was 2.3 mg L⁻¹ h⁻¹ and after 14 days the biomass contained more than 30% lipids by dry weight. To attain this, the nitrogen was provided as KNO₃ at an initial concentration of 2.05 g L⁻¹ and chelated ferric iron was added at a concentration of 1.2 × 10⁻⁵ mol L⁻¹ on the ninth day. Under the same conditions in culture tubes (36 mm outer diameter), the biomass productivity was 2.8-fold greater than in the photobioreactor (0.125 m in diameter), but the lipid productivity was only 1.2-fold higher. Thus, the average low-light level in the photobioreactor actually increased the biomass specific lipid production compared to the culture tubes. A light-limited growth model closely agreed with the experimental profiles of biomass production, nitrogen consumption, and lipid production in the photobioreactor.     

Sedimentary Organic Carbon and Nitrogen Sequestration Across a Vertical Gradient on a Temperate Wetland Seascape Including Salt Marshes,Seagrass Meadows and Rhizophytic Macroalgae Beds

Coastal wetlands are key in regulating coastal carbon and nitrogen dynamics and contribute significantly to climate change mitigation and anthropogenic nutrient reduction. We investigated organic carbon (OC) and total nitrogen (TN) stocks and burial rates at four adjacent vegetated coastal habitats across the seascape elevation gradient of Cádiz Bay (South Spain), including one species of salt marsh, two of seagrasses, and a macroalgae. OC and TN stocks in the upper 1 m sediment layer were higher at the subtidal seagrass Cymodocea nodosa (72.3 Mg OC ha⁻¹, 8.6 Mg TN ha⁻¹) followed by the upper intertidal salt marsh Sporobolus maritimus (66.5 Mg OC ha⁻¹, 5.9 Mg TN ha⁻¹), the subtidal rhizophytic macroalgae Caulerpa prolifera (62.2 Mg OC ha⁻¹, 7.2 Mg TN ha⁻¹), and the lower intertidal seagrass Zostera noltei (52.8 Mg OC ha⁻¹, 5.2 Mg TN ha⁻¹). The sedimentation rates increased from lower to higher elevation, from the intertidal salt marsh (0.24 g cm⁻² y⁻¹) to the subtidal macroalgae (0.12 g cm⁻² y⁻¹). The organic carbon burial rate was highest at the intertidal salt marsh (91 ± 31 g OC m⁻² y⁻¹), followed by the intertidal seagrass, (44 ± 15 g OC m⁻² y⁻¹), the subtidal seagrass (39 ± 6 g OC m⁻² y⁻¹), and the subtidal macroalgae (28 ± 4 g OC m⁻² y⁻¹). Total nitrogen burial rates were similar among the three lower vegetation types, ranging from 5 ± 2 to 3 ± 1 g TN m⁻² y⁻¹, and peaked at S. maritimus salt marsh with 7 ± 1 g TN m⁻² y⁻¹. The contribution of allochthonous sources to the sedimentary organic matter decreased with elevation, from 72% in C. prolifera to 33% at S. maritimus. Our results highlight the need of using habitat-specific OC and TN stocks and burial rates to improve our ability to predict OC and TN sequestration capacity of vegetated coastal habitats at the seascape level. We also demonstrated that the stocks and burial rates in C. prolifera habitats were within the range of well-accepted blue carbon ecosystems such as seagrass meadows and salt marshes.

     

Cultivation of Chlorella vulgaris in tubular photobioreactors: A lipid source for biodiesel production

Chlorella vulgaris was cultivated in two different 2.0 L-helicoidal and horizontal photobioreactors at 5 klux using the bicarbonate contained in the medium and ambient air as the main CO₂ sources. The influence of bicarbonate concentration on biomass growth as well as lipid content and profile was first investigated in shake flasks, where the stationary phase was achieved in about one half the time required by the control. The best NaHCO₃ concentration (0.2 g L⁻¹) was then used in both photobioreactors. While the fed-batch run performed in the helicoidal photobioreactor provided the best result in terms of biomass productivity, which was (84.8 mg L⁻¹ d⁻¹) about 2.5-fold that of the batch run, the horizontal configuration ensured the highest lipid productivity (10.3 mg L⁻¹ d⁻¹) because of a higher lipid content of biomass (22.8%). These preliminary results suggest that the photobioreactor configuration is a key factor either for the growth or the composition of this microalga. The lipid quality of C. vulgaris biomass grown in both photobioreactors is expected to meet the standards for biodiesel, especially in the case of the helicoidal configuration, provided that further efforts will be made to optimize the conditions for its production as a biodiesel source.     

Bicarbonate-based carbon capture and algal production system on ocean with floating inflatable-membrane photobioreactor

This study aims to develop a low-cost microalgae culture system which uses a simple closed vessel as photobioreactor to save manufacturing cost, waves for mixing to save energy cost, and high concentration of bicarbonate for carbon supply to avoid the high cost of CO₂-bubbling pipeline construction on the ocean as well as to control pH by buffering the effect of bicarbonate/carbonate. To test this idea, the alkalihalophilic cyanobacterium Euhalothece sp. was cultured with 1.0 M NaHCO₃ in small-scale floating photobioreactors (PBRs) on 10-cm-high artificial waves at first. The final biomass concentration was up to 0.91 and 1.47 g L^?1 for indoor and outdoor cultures, respectively. However, the recorded dissolved oxygen (DO) was occasionally over-saturated (> 500% of air saturation), indicating mass transfer problem. k _L a in these PBRs with different culture depth was measured then, and the results showed great variation, from 0.13 to 4.87 h^?1. At the scale of 1.0 m², this floating PBR was made with low-cost membrane and inflatable design. It was placed on the ocean surface and mixed with natural waves. Biomass concentration of 1.63 g L^?1 and productivity of 8.27 g m^?2 day^?1 were obtained in this culture. With these results, the feasibility of a low-cost microalgae culture system was proven, which could systematically reduce the cost of photobioreactor manufacturing, operating, and maintenance.     

Outdoor microalgae cultivation in airlift photobioreactor at high irradiance and temperature conditions: effect of batch and fed-batch strategies,photoinhibition, and temperature stress

The microalgae Scenedesmus abundans cultivated in five identical airlift photobioreactors (PBRs) in batch and fed-batch modes at the outdoor tropical condition. The microalgae strain S. abundans was found to tolerate high temperature (35–45 °C) and high light intensity (770–1690 µmol m^− 2 s^− 1). The highest biomass productivities were 152.5–162.5 mg L^− 1 day^− 1 for fed-batch strategy. The biomass productivity was drastically reduced due to photoinhibition effect at a culture temperature of > 45 °C. The lipid compositions showed fatty acids mainly in the form of saturated and monounsaturated fatty acids (> 80%) in all PBRs with Cetane number more than 51. The fed-batch strategies efficiently produced higher biomass and lipid productivities at harsh outdoor conditions. Furthermore, the microalgae also accumulated omega-3 fatty acid (C18:3) up to 14% (w/w) of total fatty acid at given outdoor condition.

     

A submerged culture system for rapid micropropagation of the commercially important aquarium plant, ‘Amazon sword’ (Echinodorus ‘Indian Red’)

Echinodorus ‘Indian Red’ is an underwater plant, used worldwide for aquarium ornamentation. An efficient method for in vitro propagation and plantlet acclimatization of this popular aquarium plant was standardized. Surface-disinfected shoot-tips were cultured in submerged conditions in a solid–liquid bilayer medium, consisting of an upper, liquid layer (sterile distilled water) and a lower, solid layer Murashige and Skoog (MS) basal medium supplemented with 3.0% (w/v) sucrose, 0.8% (w/v) agar-agar, and plant growth regulators (PGRs) in different combinations and concentrations. The combination of 2.5 mg L⁻¹ 6-benzylaminopurine and 1.0 mg L⁻¹ α-naphthaleneacetic acid improved the multiplication rate to a maximum of 26.8 ± 0.51 shoots per explant after 60 d of culture. The number of multiplied shoots increased with each regeneration cycle, thus from only 26.8 ± 0.51 shoots per explant (first regeneration cycle), this number increased to 33.5 ± 0.58 (second regeneration cycle), and to 38.3 ± 0.62 for the third regeneration cycle with the same medium composition. The highest number of roots (8.3 ± 0.28) per shoot was induced in the presence of 1.0 mg L⁻¹ indole-3-butyric acid, but further growth of these roots was stunted. The best rooting was achieved on PGR-free ½-strength MS medium, where 6.1 ± 0.21 roots per shoot were induced with 5.8 ± 0.35 cm length after 30 d of culture. The regenerated plantlets were successfully acclimatized to submerged underwater conditions, with 100% survival rate. The present protocol is suitable for the commercial propagation of Echinodorus ‘Indian Red’ for aquarium-industries.

     

Nutritional optimization of Arthrospira platensis for starch and Total carbohydrates production

Present study aims to optimize the production of starch and total carbohydrates from Arthrospira platensis. Growing concerns toward unprecedented environmental issues associated with plastic pollution has created a tremendous impetus to develop new biomaterials for the production of bioplastic. Starch-based biopolymers from algae serve as sustainable feedstock for thermoplastic starch production due to their abundant availability and low cost. A. platensis was cultivated in Zarrouk's medium at 32 ± 1°C and exposed to red light with a photoperiod of 12:12 hr light/dark. Growth kinetics studies showed that the maximum specific growth rate (μ_max) obtained was 0.059 day⁻¹ with the doubling time (t_d) of 11.748 days. Subsequently, Zarrouk's medium with different concentrations of sulfur, phosphorus and nitrogen was prepared to establish the nutrient-limiting conditions to enhance the accumulation of starch and total carbohydrates. In this study, the highest starch accumulated was 6.406 ± 0.622 mg L⁻¹ under optimized phosphorus limitation (0.025 g L⁻¹) conditions. Nitrogen limitation (0.250 g L⁻¹) results demonstrated significant influenced (p < 0.05) on total carbohydrates (67.573 ± 2.893 mg L⁻¹) accumulation in A. platensis. The starch accumulation in A. platensis was significantly affected (p < 0.05) by phosphorus limitation (0.0025 g L⁻¹). Subsequently, the optimized phosphorus concentration was coupled with mixotrophic cultivation to further enhance the starch accumulation. The results obtained indicated that, the starch (11.426 ± 0.314 mg L⁻¹) and carbohydrates (43.053 ± 2.986 mg L⁻¹) concentration obtained was significantly high (p < 0.05) under mixotrophic cultivation. Therefore, it shown that nutrient limitation and mixotrophic cultivation are viable strategies to enhance the accumulation of starch and total carbohydrates in A. platensis.     

Irradiance-driven 20-hydroxyecdysone production and morphophysiological changes in Pfaffia glomerata plants grown in vitro

Pfaffia glomerata possesses potential pharmacological and medicinal properties, mainly owing to the secondary metabolite 20-hydroxyecdysone (20E). Increasing production of biomass and 20E is important for industrial purposes. This study aimed to evaluate the influence of irradiance on plant morphology and production of 20E in P. glomerata grown in vitro. Nodal segments of accessions 22 and 43 (Ac22 and Ac43) were inoculated in culture medium containing MS salts and vitamins. Cultures were maintained at 25 ± 2 °C under a 16-h photoperiod and subjected to irradiance treatments of 65, 130, and 200 μmol m⁻² s⁻¹ by fluorescent lamps. After 30 days, growth parameters, pigment content, stomatal density, in vitro photosynthesis, metabolites content, and morphoanatomy were assessed. Notably, Ac22 plants exhibited 10-fold higher 20E production when cultivated at 200 μmol m⁻² s⁻¹ than at 65 μmol m⁻² s⁻¹, evidencing the importance of light quantity for the accumulation of this metabolite. 20E production was twice as high in Ac22 as in Ac43 plants although both accessions responded positively to higher irradiance. Growth under 200 μmol m⁻² s⁻¹ stimulated photosynthesis and consequent biomass accumulation, but lowered carotenoids and anthocyanins. Furthermore, increasing irradiance enhanced the number of palisade and spongy parenchyma cells, enhancing the overall growth of P. glomerata.

Graphical abstract

     

A closed solar photobioreactor for cultivation of microalgae under supra-high irradiance: basic design and performance

An account is given of the setting up and use of a novel type of closed tubular photobioreactor at the Academic and University Centre in Nove Hrady, Czech Republic. This "penthouse-roof" photobioreactor was based on solar concentrators (linear Fresnel lenses) mounted in a climate-controlled greenhouse on top of the laboratory complex combining features of indoor and outdoor cultivation units. The dual-purpose system was designed for algal biomass production in temperate climate zone under well-controlled cultivation conditions and with surplus solar energy being used for heating service water. The system was used to study the strategy of microalgal acclimation to supra-high solar irradiance, with values as much as 3.5 times the ambient value, making the approach unique. The cultivation system proved to be fully functional with sufficient mixing and cooling, efficient oxygen stripping and light tracking. Experimental results (measurement of the maximum photochemical yield of PSII and non-photochemical quenching) showed that the cyanobacterium Spirulina (= Arthrospira) platensis cultivated under sufficient turbulence and biomass density was able to acclimate to irradiance values as high as 7 mmol photon m^–2 s^–1. The optimal biomass concentration of Spirulina cultures in September ranged between 1.2 to 2.2 g L^–1, which resulted in a net productivity of about 0.5 g L^–1 d^–1 corresponding to a biomass yield of 32.5 g m^–2 d^–1 (based on the minimum illuminated surface area of the photobioreactor).     

Characteristics and molecular determinants of a highly selective and efficient glycyrrhizin-hydrolyzing β-glucuronidase from Staphylococcus pasteuri 3I10

Glycyrrhizin (GL), the principal sweet-tasting bioactive ingredient of licorice (root of Glycyrrhiza glabra), shows poor oral absorption and gut microbial transformation of GL to glycyrrhetinic acid (GA) plays a major role for its multiple pharmacological effects. Co-administration of GL-hydrolyzing bacteria appears to be a feasible strategy to enhance GA exposure. This study reported a gut bacterial strain Staphylococcus pasteuri 3I10 which exhibited moderate p-nitrophenyl-β-D-glucuronide (PNPG)-hydrolyzing activity but low GL deglucuronidation activity in its crude lysate. The gus gene encoding S. pasteuri 3I10 β-glucuronidase was successfully cloned and overexpressed in Escherichia coli BL21(DE3). The purified β-glucuronidase (SpasGUS) was 71 kDa and showed optimal pH and temperature at 6.0 and 50 °C, respectively. Comparing to E. coli β-glucuronidase (EcoGUS), SpasGUS displayed lower velocity and affinity to PNPG hydrolysis (V_max 16.1 ± 0.9 vs 140.0 ± 4.1 μmolmin⁻¹ mg⁻¹; K_m 469.4 ± 73.4 vs 268.0 ± 25.8 μM), but could selectively convert GL to GA at much higher efficiency (V_max 0.41 ± 0.011 vs 0.005 ± 0.002 μmolmin⁻¹ mg⁻¹; K_m 116.9 ± 15.4 vs 53.4 ± 34.8 μM). Molecular docking studies suggested SpasGUS formed hydrogen bond interactions with the glucuronic acids at Asn414, Glu415 and Leu450, and Val159, Tyr475, Ala368, and Phe367 provided a hydrophobic environment for enhanced activity. Two special substrate interaction loops near the binding pocket of SpasGUS (loop 1 β-glucuronidase) may account for the selective and efficient bioconversion of GL to GA, predicting that loop 1 β-glucuronidases show high possibility in processing GL than mini-loop 1 and loop 2 β-glucuronidases. These findings support potential applications of SpasGUS in cleaving GL to facilitate GA production in vivo or in pharmaceutical industry.

     

Simultaneous nutrients and carbon removal during pretreated swine slurry degradation in a tubular biofilm photobioreactor

The biodegradation potential of an innovative enclosed tubular biofilm photobioreactor inoculated with a Chlorella sorokiniana strain and an acclimated activated sludge consortium was evaluated under continuous illumination and increasing pretreated (centrifuged) swine slurry loading rates. This photobioreactor configuration provided simultaneous and efficient carbon, nitrogen, and phosphorous treatment in a single-stage process at sustained nitrogen and phosphorous removals efficiencies ranging from 94% to 100% and 70–90%, respectively. Maximum total organic carbon (TOC), NH₄ ⁺, and PO₄ ³⁻ removal rates of 80 ± 5 g C m_r ⁻³ day⁻¹, 89 ± 5 g N m_r ⁻³ day⁻¹, and 13 ± 3 g P m_r ⁻³ day⁻¹, respectively, were recorded at the highest swine slurry loadings (TOC of 1,247 ± 62 mg L⁻¹, N–NH₄ ⁺ of 656 ± 37 mg L⁻¹, P–PO₄ ³⁺ of 117 ± 19 mg L⁻¹, and 7 days of hydraulic retention time). The unusual substrates diffusional pathways established within the phototrophic biofilm (photosynthetic O₂ and TOC/NH₄ ⁺ diffusing from opposite sides of the biofilm) allowed both the occurrence of a simultaneous denitrification/nitrification process at the highest swine slurry loading rate and the protection of microalgae from any potential inhibitory effect mediated by the combination of high pH and high NH₃ concentrations. In addition, this biofilm-based photobioreactor supported efficient biomass retention (>92% of the biomass generated during the pretreated swine slurry biodegradation).     

Novel polycondensed biopolyamide generated from biomass-derived 4-aminohydrocinnamic acid

Biomass plastics are expected to contribute to the establishment of a carbon-neutral society by replacing conventional plastics derived from petroleum. The biomass-derived aromatic amine 4-aminocinnamic acid (4ACA) produced by recombinant bacteria is applied to the synthesis of high-performance biopolymers such as polyamides and polyimides. Here, we developed a microbial catalyst that hydrogenates the α,β-unsaturated carboxylic acid of 4ACA to generate 4-aminohydrocinnamic acid (4AHCA). The ability of 10 microbial genes for enoate and xenobiotic reductases expressed in Escherichia coli to convert 4ACA to 4AHCA was assessed. A strain producing 2-enoate reductase from Clostridium acetobutylicum (ca2ENR) reduced 4ACA to 4AHCA with a yield of > 95% mol mol⁻¹ and reaction rates of 3.4 ± 0.4 and 4.4 ± 0.6 mM h⁻¹ OD₆₀₀ ⁻¹ at the optimum pH of 7.0 under aerobic and anaerobic conditions, respectively. This recombinant strain reduced caffeic, cinnamic, coumaric, and 4-nitrocinnamic acids to their corresponding propanoic acid derivatives. We polycondensed 4AHCA generated from biomass-derived 4ACA by dehydration under a catalyst to form high-molecular-weight poly(4AHCA) with a molecular weight of M _n = 1.94 MDa. This polyamide had high thermal properties as indicated by a 10% reduction in weight at a temperature of T _d10 = 394 °C and a glass transition temperature of T _g = 240 °C. Poly(4AHCA) derived from biomass is stable at high temperatures and could be applicable to the production of high-performance engineering plastics.

     

ndpT encodes a new protein involved in nicotine catabolism by Sphingomonas melonis TY

Sphingomonas melonis TY utilizes nicotine as a sole source of carbon, nitrogen, and energy to grow. One of the genes in its ndp catabolic cluster, ndpT, encodes a hypothetical transporter. Since no transporter for nicotine has been identified in microorganisms, we investigated whether NdpT is responsible for nicotine transport. ndpT was induced by nicotine, and gene knockout and complementation studies clearly indicated that ndpT is essential for the catabolism of nicotine in strain TY. NdpT-GFP was located at the periphery of the cells, suggesting that NdpT is a membrane protein. Uptake assays with L-[¹⁴C] nicotine illustrated that nicotine uptake in strain TY is mediated by a constitutively synthesized permease with a K_m of 0.362 ± 0.07 μM and a V_max of 0.762 ± 0.068 μmol min⁻¹ (mg cell dry weight)⁻¹ and that ndpT may play a role in nicotine exclusion. Hence, we consider NdpT a nicotine catabolism-related protein.

     

Evaluation of the Impacts of Long-Term Enriched Artemia with Bacillus subtilis on Growth Performance,Reproduction, Intestinal Microflora,and Resistance to Aeromonas hydrophila of Ornamental Fish Poecilia latipinna

The present study investigated the effect of enriched Artemia with Bacillus subtilis on growth performance, reproductive factors, proximate composition, intestinal microflora, and resistance to Aeromonas hydrophila of ornamental fish, Poecilia latipinna. Using a completely randomized design, the experiment included three groups. The first group was fed with commercial food without any probiotic. The second group was fed with unenriched Artemia, and the last group consumed long-time enriched Artemia with Bacillus subtilis. The bacteria B. subtilis with a density of 1 × 10⁵ CFU mL⁻¹ was added daily to Artemia culture medium. The total microflora and Bacillus subtilis counts were significantly increased in enriched Artemia compared to the unenriched group (P < 0.05). In fish fed groups, growth factors did not show any significant difference (P > 0.05). The maximum relative fecundity (28.65 ± 2.52 egg number g⁻¹), fry production (62.93 ± 4.6 individual per female), and fry survival (70.97 ± 1.56%) obtained in the third group were found to be significantly more than those in the first and the second groups. Moreover, intestinal bacterial count for Bacillus revealed that the higher concentration of bacteria was significantly related to the third group (6.24 ± 0.11 log CFU g⁻¹) (P < 0.05). Maximum protein and fat contents were observed in fish fed with Bacillus-enriched Artemia; however, no significant difference was found between control and unenriched Artemia groups (P > 0.05). The highest amount of ash was observed in fish fed with commercial food without any probiotic (P < 0.05). At the end of the feeding period, each of the three groups along with positive group (oxytetracycline 100 mg kg⁻¹ of commercial food) was exposed to A. hydrophila (BCCM5/LMG3770) bacteria intraperitoneally. Based on the results, the lowest cumulative mortality was significantly found in group three (68.75 ± 3.6%) and positive group (62.5 ± 7.0%) compared to control and unenriched Artemia groups (P < 0.05). Hence, B. subtilis with a concentration of 1 × 10⁵ CFU mL⁻¹ during the period of Artemia culturing can improve the reproductive parameters, intestinal microflora, and resistance to pathogenic bacteria of Poecilia latipinna.

     

Kinetic modeling of light limitation and sulfur deprivation effects in the induction of hydrogen production with Chlamydomonas reinhardtii. Part II: Definition of model‐based protocols and experimental validation

Photosynthetic hydrogen production under light by the green microalga Chlamydomonas reinhardtii was investigated in a torus‐shaped PBR in sulfur‐deprived conditions. Culture conditions, represented by the dry biomass concentration of the inoculum, sulfate concentration, and incident photon flux density (PFD), were optimized based on a previously published model (Fouchard et al., 2009. Biotechnol Bioeng 102:232–245). This allowed a strictly autotrophic production, whereas the sulfur‐deprived protocol is usually applied in photoheterotrophic conditions. Experimental results combined with additional information from kinetic simulations emphasize effects of sulfur deprivation and light attenuation in the PBR in inducing anoxia and hydrogen production. A broad range of PFD was tested (up to 500 µmol photons m⁻² s⁻¹). Maximum hydrogen productivities were 1.0 ± 0.2 mL H₂/h/L (or 25 ± 5 mL H₂/m² h) and 3.1 mL ± 0.4 H₂/h L (or 77.5 ± 10 mL H₂/m² h), at 110 and 500 µmol photons m⁻² s⁻¹, respectively. These values approached a maximum specific productivity of approximately 1.9 mL ± 0.4 H₂/h/g of biomass dry weight, clearly indicative of a limitation in cell capacity to produce hydrogen. The efficiency of the process and further optimizations are discussed. Biotechnol. Bioeng. 2011;108: 2288–2299. © 2011 Wiley Periodicals, Inc.     

Growth and biomass productivity of Scenedesmus vacuolatus on a twin layer system and a comparison with other types of cultivations

Scenedesmus is a genus of microalgae employed for several industrial uses. Industrial cultivations are performed in open ponds or in closed photobioreactors (PBRs). In the last years, a novel type of PBR based on immobilized microalgae has been developed termed porous substrate photobioreactors (PSBR) to achieve significant higher biomass density during cultivation in comparison to classical PBRs. This work presents a study of the growth of Scenedesmus vacuolatus in a Twin Layer System PSBR at different light intensities (600 μmol photons m⁻² s⁻¹ or 1000 μmol photons m⁻² s⁻¹), different types and concentrations of the nitrogen sources (nitrate or urea), and at two CO₂ levels in the gas phase (2% or 0.04% v/v). The microalgal growth was followed by monitoring the attached biomass density as dry weight, the specific growth rate and pigment accumulation. The highest productivity (29 g m⁻² d⁻¹) was observed at a light intensity of 600 μmol photons m⁻² s⁻¹ and 2% CO₂. The types and concentrations of nitrogen sources did not influence the biomass productivity. Instead, the higher light intensity of 1000 μmol photons m⁻² s⁻¹ and an ambient CO₂ concentration (0.04%) resulted in a significant decrease of productivity to 18 and 10–12 g m⁻² d⁻¹, respectively. When compared to the performance of similar cultivation systems (15–30 g m⁻² d⁻¹), these results indicate that the Twin Layer cultivation System is a competitive technique for intensified microalgal cultivation in terms of productivity and, at the same time, biomass density.

     

Morphophysiological in vitro performance of Brazilian ginseng (Pfaffia glomerata (Spreng.) Pedersen) based on culture medium formulations

Pfaffia glomerata has potential pharmacological and medicinal properties due to the production of a secondary metabolite known as the phytoecdysteroid 20-hydroxyecdysone (20E). There have been increasing efforts for massive in vitro propagation of Pfaffia plants due to high extractivism and overharvesting of this species. Research on the species has shown that photoautotrophic cultivation can improve the production of 20E. In addition, other abiotic factors such as the formulations of culture media can influence the morphophysiological behavior of the plants in vitro. Therefore, the objective of this study was to analyze the morphological and physiological performances of P. glomerata plants in different formulations of culture media, under photoautotrophic and photomixotrophic propagation conditions. Six medium formulations, the Driver and Kuniyuki medium (DKW), Correia et al. medium (JADS), Murashige and Skoog medium (MS), Quoirin and Lepoivre medium (QL), Rugini medium (OM), and Woody Plant medium (WPM), all supplemented with DKW vitamins, 100 mg L⁻¹ myo-inositol, 6.5 g L⁻¹ agar, and with or without 3% (w/v) sucrose, were evaluated. Cultures were maintained at 25 ± 2°C, with a 16 h-photoperiod under 60 μmol m⁻² s⁻¹ of irradiance under a fluorescent lamp for 50 d. Results showed that the presence or absence of sucrose, and the different nutritional formulations influenced growth, photosynthetic pigment content, endogenous levels of sugars, leaf morphology, levels of 20E, and transport of water and minerals in P. glomerata. Notably, OM, DKW, QL, and WPM media promoted higher production of 20E under photomixotrophic growth conditions.

     

Innovative development of membrane sparger for carbon dioxide supply in microalgae cultures

The present study was aimed to develop a membrane sparger (MS) integrated into a tubular photobioreactor to promote the increase of the carbon dioxide (CO₂) fixation by Spirulina sp. LEB 18 cultures. The use of MS for the CO₂ supply in Spirulina cultures resulted not only in the increase of DIC concentrations but also in the highest accumulated DIC concentration in the liquid medium (127.4 mg L⁻¹ d⁻¹). The highest values of biomass concentration (1.98 g L⁻¹), biomass productivity (131.8 mg L⁻¹ d⁻¹), carbon in biomass (47.9% w w⁻¹), CO₂ fixation rate (231.6 mg L⁻¹ d⁻¹), and CO₂ use efficiency (80.5% w w⁻¹) by Spirulina were verified with MS, compared to the culture with conventional sparger for CO₂ supply. Spirulina biomass in both culture conditions had high protein contents varying from 64.9 to 69% (w w⁻¹). MS can be considered an innovative system for the supply of carbon for the microalgae cultivation and biomass production. Moreover, the use of membrane system might contribute to increased process efficiency with a reduced cost of biomass production.