The LiCl effect on the conformation of lentinan in DMSO

Lentinan (β‐(1→3)‐D ‐glucan) was found to be successfully fractionated by the mixture of dimethyl sulfoxide (DMSO) and lithium chloride (LiCl) as a solvent and acetone as a precipitant. Light scattering and viscosity measurements were made on solutions of fractionated samples in pure DMSO and 0.2M LiCl/DMSO in the range of the molecular weight M_w from 21.7 × 10⁴ to 84.7 × 10⁴. The values of M_w in both solvents were almost the same, but the remarkable difference between the values of intrinsic viscosity [η] demonstrated that the LiCl/DMSO solvent greatly enhances the stiffness of the lentinan backbone. The observed intrinsic viscosity [η] was analyzed by the Yoshizaki‐Nitta‐Yamakawa theory of a worm‐like chain, and the persistence length q and molecular weight per unit contour length M_L were determined roughly as 6.0 nm and 890 g nm^?1 in 0.2M LiCl/DMSO, and 5.1 nm and 890 g nm^?1 in pure DMSO, respectively. This slightly larger persistent length in 0.2M LiCl/DMSO also confirmed the higher stiffness of lentinan enhanced by the LiCl/DMSO solvent. The enhancement of the chain stiffness was ascribed to the electrostatic repulsion because of the hydrogen bonding of the hydroxyl protons of lentinan with the chloride ion, which is in turn associated with the Li⁺(DMSO)_n macrocation complex. © 2012 Wiley Periodicals, Inc. Biopolymers 97: 840–845, 2012.     

Outstanding Low Temperature Thermoelectric Power Factor from Completely Organic Thin Films Enabled by Multidimensional Conjugated Nanomaterials

In an effort to create a paintable/printable thermoelectric material, comprised exclusively of organic components, polyaniline (PANi), graphene, and double‐walled nanotube (DWNT) are alternately deposited from aqueous solutions using the layer‐by‐layer assembly technique. Graphene and DWNT are stabilized with an intrinsically conductive polymer, poly(3,4‐ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS). An 80 quadlayer thin film (≈1 μm thick), comprised of a PANi/graphene‐PEDOT:PSS/PANi/DWNT‐PEDOT:PSS repeating sequence, exhibits unprecedented electrical conductivity (σ ≈ 1.9 × 10⁵ S m^?1) and Seebeck coefficient (S ≈ 120 μV K^?1) for a completely organic material. These two values yield a thermoelectric power factor (PF = S ² σ ^?1) of 2710 μW m^?1 K^?2, which is the highest value ever reported for a completely organic material and among the highest for any material measured at room temperature. These outstanding properties are attributed to the highly ordered structure in the multilayer assembly. This water‐based thermoelectric nanocomposite is competitive with the best inorganic semiconductors (e.g., bismuth telluride) at room temperature and can be applied as a coating to any flexible surface (e.g., fibers in clothing). For the first time, there is a real opportunity to harness waste heat from unconventional sources, such as body heat, to power devices in an environmentally‐friendly way.     

Recombinant Protein Production with Pichia pastoris in Continuous Fermentation – Kinetic Analysis of Growth and Product Formation

Continuous fermentation was applied to the production of recombinant human chymotrypsinogen B (hCTRB) by the methylotrophic yeast Pichia pastoris as a tool for the kinetic analysis of growth and product formation. Using methanol as the sole source of carbon, energy, and induction, cell growth could be described by a non‐competitive Monod approach. Maximum growth rate μ_max was determined to 0.084 h^‐‐1 and the K_M‐value for methanol to 0.22 g·L^‐‐1, respectively. With respect to product formation, a similar model was established exhibiting a methanol concentration of 0.13 g·L^‐‐1 as the K_M‐value and a maximum biomass‐specific product‐formation rate of π_max = 0.23 mg·g^‐‐1·h^‐‐1. The production of hCTRB was strictly growth‐coupled. The data provided covers the range of methanol concentrations between 0 and 4 g·L^‐‐1. Substrate concentrations exceeding this upper value led to a complete collapse of product formation. This change in phenotype turned out to be irreversible indicating a genetic instability of transformed Pichia pastoris caused by excess methanol.     

Soil carbon changes under Miscanthus driven by C4 accumulation and C3 decompostion – toward a default sequestration function

Bioenergy has to meet increasing sustainability criteria in the EU putting conventional bioenergy crops under pressure. Alternatively, perennial bioenergy crops, such as Miscanthus, show higher greenhouse gas savings with similarly high energy yields. In addition, Miscanthus plantations may sequester additional soil organic carbon (SOC) to mitigate climate change. As the land‐use change in cropland to Miscanthus involves a C₃‐C₄ vegetation change (VC), it is possible to determine the dynamic of Miscanthus‐derived SOC (C₄ carbon) and of the old SOC (C₃ carbon) by the isotopic ratio of ¹³C to ¹²C. We sampled six croplands and adjacent Miscanthus plantations exceeding the age of 10 years across Europe. We found a mean C₄ carbon sequestration rate of 0.78 ± 0.19 Mg ha^?1 yr^?1, which increased with mean annual temperature. At three of six sites, we found a significant increase in C₃ carbon due to the application of organic fertilizers or difference in baseline SOC, which we define as non‐VC‐induced SOC changes. The Rothamsted Carbon Model was used to disentangle the decomposition of old C₃ carbon and the non‐VC‐induced C₃ carbon changes. Subsequently, this method was applied to eight more sites from the literature, resulting in a climate‐dependent VC‐induced SOC sequestration rate (0.40 ± 0.20 Mg ha^?1 yr^?1), as a step toward a default SOC change function for Miscanthus plantations on former croplands in Europe. Furthermore, we conducted a SOC fractionation to assess qualitative SOC changes and the incorporation of C₄ carbon into the soil. Sixteen years after Miscanthus establishment, 68% of the particulate organic matter (POM) was Miscanthus‐derived in 0–10 cm depth. POM was thus the fastest cycling SOC fraction with a C₄ carbon accumulation rate of 0.33 ± 0.05 Mg ha^?1 yr^?1. Miscanthus‐derived SOC also entered the NaOCl‐resistant fraction, comprising 12% in 0–10 cm, which indicates that this fraction was not an inert SOC pool.     

Optimization of norovirus virus‐like particle production in Pichia pastoris using a real‐time near‐infrared bioprocess monitor

The production of norovirus virus‐like particles (NoV VLPs) displaying NY‐ESO‐1 cancer testis antigen in Pichia pastoris BG11 Mut⁺ has been enhanced through feed‐strategy optimization using a near‐infrared bioprocess monitor (RTBio^® Bioprocess Monitor, ASL Analytical, Inc.), capable of monitoring and controlling the concentrations of glycerol and methanol in real‐time. The production of NoV VLPs displaying NY‐ESO‐1 in P. pastoris has potential as a novel cancer vaccine platform. Optimization of the growth conditions resulted in an almost two‐fold increase in the expression levels in the fermentation supernatant of P. pastoris as compared to the starting conditions. We investigated the effect of methanol concentration, batch phase time, and batch to induction transition on NoV VLP‐NY‐ESO‐1 production. The optimized process included a glycerol transition phase during the first 2 h of induction and a methanol concentration set point of 4 g L^?1 during induction. Utilizing the bioprocess monitor to control the glycerol and methanol concentrations during induction resulted in a maximum NoV VP1‐NY‐ESO‐1 yield of 0.85 g L^?1. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:518–526, 2016     

Quantitative physiology of Pichia pastoris during glucose‐limited high‐cell density fed‐batch cultivation for recombinant protein production

Pichia pastoris has become one of the major microorganisms for the production of proteins in recent years. This development was mainly driven by the readily available genetic tools and the ease of high‐cell density cultivations using methanol (or methanol/glycerol mixtures) as inducer and carbon source. To overcome the observed limitations of methanol use such as high heat development, cell lysis, and explosion hazard, we here revisited the possibility to produce proteins with P. pastoris using glucose as sole carbon source. Using a recombinant P. pastoris strain in glucose limited fed‐batch cultivations, very high‐cell densities were reached (more than 200 g_CDW L^?1) resulting in a recombinant protein titer of about 6.5 g L^?1. To investigate the impact of recombinant protein production and high‐cell density fermentation on the metabolism of P. pastoris, we used ¹³C‐tracer‐based metabolic flux analysis in batch and fed‐batch experiments. At a controlled growth rate of 0.12 h^?1 in fed‐batch experiments an increased TCA cycle flux of 1.1 mmol g^?1 h^?1 compared to 0.7 mmol g^?1 h^?1 for the recombinant and reference strains, respectively, suggest a limited but significant flux rerouting of carbon and energy resources. This change in flux is most likely causal to protein synthesis. In summary, the results highlight the potential of glucose as carbon and energy source, enabling high biomass concentrations and protein titers. The insights into the operation of metabolism during recombinant protein production might guide strain design and fermentation development. Biotechnol. Bioeng. 2010;107: 357–368. © 2010 Wiley Periodicals, Inc.     

Organic geochemical studies of modern microbial mats from Shark Bay: Part I: Influence of depth and salinity on lipid biomarkers and their isotopic signatures

The present study investigated the influence of abiotic conditions on microbial mat communities from Shark Bay, a World Heritage area well known for a diverse range of extant mats presenting structural similarities with ancient stromatolites. The distributions and stable carbon isotopic values of lipid biomarkers [aliphatic hydrocarbons and polar lipid fatty acids (PLFAs)] and bulk carbon and nitrogen isotope values of biomass were analysed in four different types of mats along a tidal flat gradient to characterize the microbial communities and systematically investigate the relationship of the above parameters with water depth. Cyanobacteria were dominant in all mats, as demonstrated by the presence of diagnostic hydrocarbons (e.g. n‐C₁₇ and n‐C_17:1). Several subtle but important differences in lipid composition across the littoral gradient were, however, evident. For instance, the shallower mats contained a higher diatom contribution, concordant with previous mat studies from other locations (e.g. Antarctica). Conversely, the organic matter (OM) of the deeper mats showed evidence for a higher seagrass contribution [high C/N, ¹³C‐depleted long‐chain n‐alkanes]. The morphological structure of the mats may have influenced CO₂ diffusion leading to more ¹³C‐enriched lipids in the shallow mats. Alternatively, changes in CO₂ fixation pathways, such as increase in the acetyl COA‐pathway by sulphate‐reducing bacteria, could have also caused the observed shifts in δ¹³C values of the mats. In addition, three smooth mats from different Shark Bay sites were analysed to investigate potential functional relationship of the microbial communities with differing salinity levels. The C_25:1 HBI was identified in the high salinity mat only and a lower abundance of PLFAs associated with diatoms was observed in the less saline mats, suggesting a higher abundance of diatoms at the most saline site. Furthermore, it appeared that the most and least saline mats were dominated by autotrophic biomass using different CO₂ fixation pathways.     

Chiral separation of flurbiprofen enantiomers by preparative and simulated moving bed chromatography

This study presents the chiral resolution of flurbiprofen enantiomers by preparative liquid chromatography using the simulated moving bed (SMB) technology. Flurbiprofen enantiomers are widely used as nonsteroidal anti‐inflammatory drugs, and although demonstrate different therapeutic actions, they are still marketed as a racemic mixture. The results presented here clearly show the importance of the selection of the proper solvent composition for the preparative separation of flurbiprofen enantiomers. Chiral SMB separation is carried out using a laboratory‐scale unit (the FlexSMB‐LSRE®) with six columns, packed with the Chiralpak AD® stationary phase (20 μm). Results presented include the experimental measurement of equilibrium and kinetic data for two very different solvent compositions, a traditional high hydrocarbon content [10%ethanol/90%n‐hexane/0.01% trifluoroacetic acid (TFA)] and a strong polar organic composition (100%ethanol/0.01%TFA). Experimental data, obtained using the two mobile phase compositions, are used to predict and optimize the SMB operation. After selecting 10%ethanol/90%n‐hexane/0.01%TFA as the most appropriate solvent composition, three feed concentrations of racemic flurbiprofen were considered. Using 40 g/l of racemic flurbiprofen feed solution, the purities for both outlet streams were above 99.4%, the productivity was 13.1 g_feed/(L_bed h), and a solvent consumption of 0.41 L_solvent/g_feed was achieved. Chirality, 2011. © 2011 Wiley‐Liss, Inc.     

Utilization of inorganic carbon in the edible cyanobacterium Ge-Xian-Mi (Nostoc) and its role in alleviating photo-inhibition

The present work investigated the inorganic carbon (C_i) uptake, fluorescence quenching and photo‐inhibition of the edible cyanobacterium Ge‐Xian‐Mi (Nostoc) to obtain an insight into the role of CO₂ concentrating mechanism (CCM) operation in alleviating photo‐inhibition. Ge‐Xian‐Mi used HCO₃^– in addition to CO₂ for its photosynthesis and oxygen evolution was greater than the theoretical rates of CO₂ production derived from uncatalysed dehydration of HCO₃^–. Multiple transporters for CO₂ and HCO₃^– operated in air‐grown Ge‐Xian‐Mi. Na⁺‐dependent HCO₃^– transport was the primary mode of active C_i uptake and contributed 53–62% of net photosynthetic activity at 250 µmol L^?1 KHCO₃ and pH 8.0. However, the CO₂‐uptake systems and Na⁺‐independent HCO₃^– transport played minor roles in Ge‐Xian‐Mi and supported, respectively, 39 and 8% of net photosynthetic activity. The steady‐state fluorescence decreased and the photochemical quenching increased in response to the transport‐mediated accumulation of intracellular C_i. Inorganic carbon transport was a major factor in facilitating quenching during the initial stage and the initial rate of fluorescence quenching in the presence of iodoacetamide, an inhibitor of CO₂ fixation, was 88% of control. Both the initial rate and extent of fluorescence quenching increased with increasing external dissolved inorganic carbon (DIC) and saturated at higher than 200 µmol L^?1 HCO₃^–. The operation of the CCM in Ge‐Xian‐Mi served as a means of diminishing photodynamic damage by dissipating excess light energy and higher external DIC in the range of 100–10000 µmol L^?1 KHCO₃ was associated with more severe photo‐inhibition under strong irradiance.     

Syngas fermentation to biofuel: Evaluation of carbon monoxide mass transfer coefficient (kLa) in different reactor configurations

Lignocellulosic biomass such as agri‐residues, agri‐processing by‐products, and energy crops do not compete with food and feed, and is considered to be the ideal renewable feedstocks for biofuel production. Gasification of biomass produces synthesis gas (syngas), a mixture primarily consisting of CO and H₂. The produced syngas can be converted to ethanol by anaerobic microbial catalysts especially acetogenic bacteria such as various clostridia species.One of the major drawbacks associated with syngas fermentation is the mass transfer limitation of these sparingly soluble gases in the aqueous phase. One way of addressing this issue is the improvement in reactor design to achieve a higher volumetric mass transfer coefficient (k_La). In this study, different reactor configurations such as a column diffuser, a 20‐μm bulb diffuser, gas sparger, gas sparger with mechanical mixing, air‐lift reactor combined with a 20‐μm bulb diffuser, air‐lift reactor combined with a single gas entry point, and a submerged composite hollow fiber membrane (CHFM) module were employed to examine the k_La values. The k_La values reported in this study ranged from 0.4 to 91.08 h^?1. The highest k_La of 91.08 h^?1 was obtained in the air‐lift reactor combined with a 20‐μm bulb diffuser, whereas the reactor with the CHFM showed the lowest k_La of 0.4 h^?1. By considering both the k_La value and the statistical significance of each configuration, the air‐lift reactor combined with a 20‐μm bulb diffuser was found to be the ideal reactor configuration for carbon monoxide mass transfer in an aqueous phase. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2011     

Simultaneous determination of three naphthoquinones in the leaves of Impatiens balsamina L. by reversed-phase high-performance liquid chromatography

Introduction – Naphthoquinones; lawsone ( 1 ), lawsone methyl ether ( 2 ) and methylene‐3,3′‐bilawsone ( 3 ) are the main active compounds of Impatiens balsamina leaves. Objective – To develop and validate an HPLC method for simultaneous quantitative determination of 1 – 3 in I. balsamina leaf extracts. Methodology – The method utilised a Supelco® C₁₈ column (5 µm, 4.6 × 150 mm) at 25°C with the mixture of 2% aqueous acetic acid : methanol (gradient elution as follows: 0–10 min, 25 : 75; 10–20 min, 32 : 68; 20–35 min, 55 : 45) as the mobile phase at a flow‐rate of 1 mL/min, and UV detection at 280 nm. The parameters of linearity, repeatability, reproducibility, accuracy specificity and sensitivity of the method were evaluated. Results – The recovery of the method was 96–101% and linearity (r² ≥ 0.9995) was obtained for all naphthoquinones. A high degree of specificity, as well as repeatability and reproducibility (RSD less than 5%), were also achieved. Copyright © 2010 John Wiley & Sons, Ltd.     

Effects of 2‐phenoxyethanol anaesthesia on juvenile meagre (Argyrosomus regius)

This study was performed to evaluate the effective concentration of the anaesthetic 2‐phenoxyethanol (2‐PE) on juvenile (1.3 ± 0.03 g) meagre (Argyrosomus regius, Asso, 1801) and establish the LC₅₀ (through a series of exposure concentrations) and LT₅₀ of 2‐PE at 20 ± 0.5°C, salinity 38 g × L^?1, pH 8.2–8.4 and dissolved oxygen >7 mg × L^?1. The induction time decreased and the recovery time increased with increasing concentrations. Conflicting results were found only in recovery time and there were no significant differences among the recovery times from all concentrations. The most suitable concentration of 2‐PE was 0.3 ml × L^?1 for about or over 15 min exposure time. The LC₅₀ and LT₅₀ for the 3–60 min exposure periods were estimated for juvenile meagre. The toxic effect of 2‐PE on survival rates of A. regius juveniles increased depending on the exposure period. In addition, 2‐phenoxyethanol LT₅₀ (median survival time) values, slope function (S) and lower and upper 95% confidence limits were estimated.     

3D‐Printed Stretchable Micro‐Supercapacitor with Remarkable Areal Performance

While stretchable micro‐supercapacitors (MSCs) have been realized, they have suffered from limited areal electrochemical performance, thus greatly restricting their practical electronic application. Herein, a facile strategy of 3D printing and unidirectional freezing of a pseudoplastic nanocomposite gel composed of Ti₃C₂T_x MXene nanosheets, manganese dioxide nanowire, silver nanowires, and fullerene to construct intrinsically stretchable MSCs with thick and honeycomb‐like porous interdigitated electrodes is introduced. The unique architecture utilizes thick electrodes and a 3D porous conductive scaffold in conjunction with interacting material properties to achieve higher loading of active materials, larger interfacial area, and faster ion transport for significantly improved areal energy and power density. Moreover, the oriented cellular scaffold with fullerene‐induced slippage cell wall structure prompts the printed electrode to withstand large deformations without breaking or exhibiting obvious performance degradation. When imbued with a polymer gel electrolyte, the 3D‐printed MSC achieves an unprecedented areal capacitance of 216.2 mF cm^?2 at a scan rate of 10 mV s^?1, and remains stable when stretched up to 50% and after 1000 stretch/release cycles. This intrinsically stretchable MSC also exhibits high rate capability and outstanding areal energy density of 19.2 µWh cm^?2 and power density of 58.3 mW cm^?2, outperforming all reported stretchable MSCs.     

Toward a High‐Performance All‐Plastic Full Battery with a'Single Organic Polymer as Both Cathode and Anode

The design and fabrication of high‐performance all‐plastic batteries is essentially important to achieve future flexible electronics. A major challenge in this field is the lack of stable and reliable soft organic electrodes with satisfactory performance. Here, a novel all‐plastic‐electrode based Li‐ion battery with a single flexible bi‐functional ladderized heterocyclic poly(quinone), (C₆O₂S₂)_n, as both cathode and anode is demonstrated. Benefiting from its unique ladder‐like quinone and dithioether structure, the as‐prepared polymer cathode shows a high energy density of 624 Wh kg^?1 (vs lithium anode) and a stable battery life of 1000 cycles. Moreover, the as‐fabricated symmetric full‐battery delivers a large capacity of 249 mAh g^?1 (at 20 mA g^?1), a good capacity retention of 119 mAh g^?1 after 250 cycles (at 1.0 A g^?1) and a noteworthy energy density up to 276 Wh kg^?1. The superior performance of poly(2,3‐dithiino‐1,4‐benzoquinone)‐based electrode rivals most of the state‐of‐the‐art demonstrations on organic‐based metal‐ion shuttling batteries. The study provides an effective strategy to develop stable bi‐functional electrode materials toward the next‐generation of high performance all‐plastic batteries.     

Layer‐by‐Layer Na3V2(PO4)3 Embedded in Reduced Graphene Oxide as Superior Rate and Ultralong‐Life Sodium‐Ion Battery Cathode

Na₃V₂(PO₄)₃ (NVP) is regarded as a promising cathode for advanced sodium‐ion batteries (SIBs) due to its high theoretical capacity and stable sodium (Na) super ion conductor (NASICON) structure. However, strongly impeded by its low electronic conductivity, the general NVP delivers undesirable rate capacity and fails to meet the demands for quick charge. Herein, a novel and facile synthesis of layer‐by‐layer NVP@reduced graphene oxide (rGO) nanocomposite is presented through modifying the surface charge of NVP gel precursor. The well‐designed layered NVP@rGO with confined NVP nanocrystal in between rGO layers offers high electronic and ionic conductivity as well as stable structure. The NVP@rGO nanocomposite with merely ≈3.0 wt% rGO and 0.5 wt% amorphous carbon, yet exhibits extraordinary electrochemical performance: a high capacity (118 mA h g^?1 at 0.5 C attaining the theoretical value), a superior rate capability (73 mA h g^?1 at 100 C and even up to 41 mA h g^?1 at 200 C), ultralong cyclability (70.0% capacity retention after 15 000 cycles at 50 C), and stable cycling performance and excellent rate capability at both low and high operating temperatures. The proposed method and designed layer‐by‐layer active nanocrystal@rGO strategy provide a new avenue to create nanostructures for advanced energy storage applications.     

Drastic Layer‐Number‐Dependent Activity Enhancement in Photocatalytic H2 Evolution over nMoS2/CdS (n ≥ 1) Under Visible Light

Exploiting noble‐metal‐free cocatalysts is of huge interest for photocatalytic water splitting using solar energy. As an efficient cocatalyst in photocatalysis, MoS₂ is shown promise as a low‐cost alternative to Pt for hydrogen evolution. Here we report a systematical study on controlled synthesis of MoS₂ with layer number ranging from ≈1 to 112 and their activities for photocatalytic H₂ evolution over commercial CdS. A drastic increase in photocatalytic H₂ evolution is observed with decreasing MoS₂ layer number. Particularly for the single‐layer (SL) MoS₂, the SL‐MoS₂/CdS sample reaches a high H₂ generation rate of ≈2.01 × 10^?3m h^?1 in Na₂S–Na₂SO₃ solutions and ≈2.59 × 10^?3m h^?1 in lactic acid solutions, corresponding to an apparent quantum efficiency of 30.2% and 38.4% at 420 nm, respectively. In addition to the more exposed edges and unsaturated active S atoms, valence band–XPS and Mott–Schottky plots analysis indicate that the SL MoS₂ has the more negative conduction band energy level than the H⁺/H₂ potential, facilitating the hydrogen reduction.     

RESPONSE TO SALINITY AND HEAT STRESS IN TWO HALOTOLERANT CHLOROPHYTE ALGAE1

Two new isolates of halotolerant chlorophyte algae from the Salt Plains National Wildlife Refuge in Oklahoma, USA, tentatively identified as Dunaliella sp. Teodoresco and Nannochloris sp. Naumann, were characterized with respect to interaction between growth salinity and short‐term heat tolerance. Cells were cultured at 23–25° C over a wide range of salinity. In both species, salinity alone had little effect on maximum photochemical yield (measured by pulse modulated fluorescence) and integrity of the light harvesting system (77 K fluorescence emission spectra). In contrast, Nannochloris exhibited decreasing growth rate (μ), light‐saturated photosynthetic capacity (P^cell_max), respiration (R_d), light‐harvesting efficiency (α^cell), and chl content with increasing salinity. Cultures were heated for 2 h near their upper temperature limits (41.5° C for Dunaliella and 45° C for Nannochloris grown at 50 psu). Dunaliella was progressively more heat‐tolerant with increasing salinity. Photochemical yield of cells at 100 and 50 psu was inhibited by about 15% and 40%, respectively, and largely recovered within 30 min after return to 23° C. Thermal inhibition of photochemical yield in Nannochloris was about 45% at both 50 and 100 psu, but recovery was slower at 100 psu. At 20 psu, both species were almost 90% inhibited by high temperature and required more than a day to recover. In both species, 2 h of heating increased the PSI:PSII fluorescence emission ratio (714:690 nm) at all salinities. This ratio largely recovered within 24 h in Dunaliella at 50 and 100 psu and partially recovered in Nannochloris at 100 psu, but cells of both species heated at 20 psu were chlorotic the next day.     

Potential agronomic options for energy-efficient sugar beet-based bioethanol production in northern Japan

Sugar beet (Beta vulgaris L. subsp. vulgaris) is deemed to be one of the most promising bioethanol feedstock crops in northern Japan. To establish viable sugar beet‐based bioethanol production systems, energy‐efficient protocols in sugar beet cultivation are being intensively sought. On this basis, the effects of alternative agronomic practices for sugar beet production on total energy inputs (from fuels and agricultural materials during cultivation and transportation) and ethanol yields (estimated from sugar yields) were assessed in terms of (i) direct drilling, (ii) reduced tillage (no moldboard plowing), (iii) no‐fungicide application, (iv) using a high‐yielding beet genotype, (v) delayed harvesting and (vi) root+crown harvesting. Compared with the conventional sugar beet production system used in the Tokachi region of Hokkaido, northern Japan, which makes use of transplants, direct drilling and no‐fungicide application contributed to reduced energy inputs from raising seedlings and fungicides, respectively, but sugar (or ethanol) yields were also reduced by these practices, to a greater equivalent extent than the reductions in energy inputs. Consequently, direct drilling (6.84 MJ L^?1) and no‐fungicide application (7.78 MJ L^?1) worsened the energy efficiency (total energy inputs to produce 1 L of ethanol), compared with conventional sugar beet production practices (5.82 MJ L^?1). Sugar yields under conventional plow‐based tillage and reduced tillage practices were similar, but total energy inputs were reduced as a result of reduced fuel consumption from not plowing. Hence, reduced tillage showed improved energy efficiency (5.36 MJ L^?1). The energy efficiency was also improved by using a high‐yielding genotype (5.23 MJ L^?1) and root+crown harvesting (5.21 MJ L^?1). For these practices, no major changes in total energy inputs were noted, but sugar yields were consistently increased. Neither total energy inputs nor ethanol yields were affected by extending the vegetative growing period by delaying harvesting.     

Production of Metarhizium anisopliae spores using nutrient‐impregnated membranes and its economic analysis

Metarhizium anisopliae spores were produced on nutrient‐impregnated membranes (NIMs). The NIM system involved wetting the membrane with a spore and nutrient suspension, followed by harvesting the spores produced after incubation. The cost efficiency of spore production was assessed for a range of nutrient sources and membrane types. Skim milk powder (20 g l^‐1) was found to be the most cost‐effective nutrient source of the nine nutrients examined. Yield was 5.7 × 10⁶ spores/cm² after 28 days incubation on a paper membrane. Supplementation of the skim milk with either sucrose (2 g l^‐1) or dextrose + KNO₃ maximized yield. Superwipe, an absorbent fibrous material, was the most efficient of 16 membranes tested which ranged from fibreglass mesh to paper and cloth. A series of small pilot plants were built, but the cost efficiency of spore production decreased as the size of the membrane increased from 24 × 24 cm to 270 × 15 cm and up to 100 × 80 cm. Yield on the two smaller pilot plants was over 10⁷spores/cm², but the cost (nutrient and membrane only) of producing 10¹³ spores (standard dose required per hectare) was around $A37 and was found not to be competitive with spore production on grain.