布朗葡萄藻的培养基选择及其产物代谢规律

通过对布朗葡萄藻分别在Chu13、Chu13×2和BG-11培养基中培养结果的比较,发现在气升式光照生物反应器中Chu13培养基最有利于布朗葡萄藻的生长和烃的合成,培养15d后,其生物量和粗烃质量分数分别为1.82g/L和58.7%;棕榈酸、油酸和亚麻酸是布朗葡萄藻的主要脂肪酸组成,Chu13培养获得的藻体不饱和脂肪酸比例最高。Chu13培养基中布朗葡萄藻代谢规律的研究表明:粗烃含量随着生物量的增加而逐渐增大,15d后粗烃产量达到最大值1.07g/L,不同生长周期烃的组成保持一致,布朗葡萄藻的烃主要由C33H56和C34H58组成;在布朗葡萄藻生长周期中,不饱和脂肪酸的比例显著上升,培养15d达到64%以上。     

Influence of CO2 on growth and hydrocarbon production in Botryococcus braunii

Botryococcus braunii is a green colonial fresh water microalga and it is recognized as one of the renewable resources for production of liquid hydrocarbons. CFTRI-Bb-1 and CFTRI-Bb-2 have been reported for the first time and their performance with regard to growth and biochemical profile is presented here. The present study focused on effect of carbon dioxide (CO2) on biomass, hydrocarbon, carbohydrate production, fatty acid profile, and carotenoid content in various species of B. braunii (LB-572, SAG 30.81, MCRC-Bb, N-836, CFTRI-Bb-1, and CFTRI-Bb-2) at 0.5, 1.0, and 2.0% (v/v) levels using a two-tier flask. CO2 at 2.0% (v/v) level enhanced growth of the organism, and a two-fold increase in biomass and carotenoid contents was observed in all the B. braunii strains studied compared with control culture (without CO2 supplementation). At 1% and 2% (v/v) CO2 concentrations, palmitic acid and oleic acid levels increased by 2.5 to 3 folds in one of the strains of B. braunii (LB-572). Hydrocarbon content was found to be above 20% at 2% CO2 level in the B. braunii LB-572, CFTRI-Bb-2, CFTRI-Bb-1, and N-836 strains, whereas it was less than 20% in the SAG 30.81 and MCRC-Bb strains compared with control culture. This culture methodology will provide information on CO2 requirement for growth of algae and metabolite production. B. braunii spp. can be grown at the tested levels of CO2 concentration without much influence on culture pH.     

Botryococcus braunii: a renewable source of hydrocarbons and other chemicals

Botryococcus braunii, a green colonial microalga, is an unusually rich renewable source of hydrocarbons and other chemicals. Hydrocarbons can constitute up to 75% of the dry mass of B. braunii. This review details the various facets of biotechnology of B. braunii, including its microbiology and physiology; production of hydrocarbons and other compounds by the alga; methods of culture; downstream recovery and processing of algal hydrocarbons; and cloning of the algal genes into other microorganisms. B. braunii converts simple inorganic compounds and sunlight to potential hydrocarbon fuels and feedstocks for the chemical industry. Microorganisms such as B. braunii can, in the long run, reduce our dependence on fossil fuels and because of this B. braunii continues to attract much attention.     

富营养废水中碳氮磷浓度对布朗葡萄藻总脂和总烃的影响

以经过二次过滤的富营养化鱼塘养殖污水为培养液,添加外源的碳、氮、磷元索,研究了污水中不同的外源无机碳、总氮和总磷浓度对布朗葡萄藻(Botryococcus braunii)生物量、总脂和总烃含量的影响.结果表明:(1)以NaHCO3作为碳源,布朗葡萄藻的生物量和总脂含量在外源无机碳浓度为5～10 mg/L时最高,总烃含量在外源无机碳浓度为15mg/L时最高.(2)以KNO3作为氮源,布朗葡萄藻的生物量在总氮浓度为15mg/L时最高,总脂含量在总氮浓度为2mg/L时最高,总烃含量在总氮浓度为20mg/L时最高.(3)以KH2 PO4作为磷源,布朗葡萄藻生物量在总磷浓度为2mg/L时最高,总脂含量和总烃含量在总磷浓度为1.5 mng/L时最高.     

Effects of carbon source, phosphorus concentration, and several micronutrients on biomass and geosmin production by Streptomyces halstedii

The effects of various carbon sources, phosphorus concentration, and different concentrations of the micronutrients calcium, cobalt, copper, iron, manganese, potassium, and zinc were determined on biomass dry weight production, geosmin production, and geosmin/biomass (G/B) values for Streptomyces halstedii, a geosmin-producing actinomycete isolated from the sediment of an aquaculture pond. Of the substrates tested, maltose as a sole carbon source promoted maximal growth by S. halstedii while mannitol promoted maximal geosmin production, and galactose yielded the highest G/B values. Fish-food pellets and galactose were poor substrates for growth. Increasing phosphorus concentrations enhanced geosmin production and G/B values. Of the seven micronutrients tested, zinc, iron, and copper had the most profound effects on biomass and geosmin production. Increasing zinc concentrations promoted biomass production while inhibiting geosmin production and G/B values; increasing concentrations of copper and iron inhibited biomass and geosmin production. Increased copper concentrations had the greatest effect in preventing growth and geosmin production by S. halstedii. Journal of Industrial Microbiology & Biotechnology (2001) 26, 241–247. Received 20 September 2000/ Accepted in revised form 17 January 2001     

Effect of nutrients on the growth of a new alpine strain of Haematococcus (Chlorophyceae) from New Zealand

The microalga Haematococcus lacustris is a source of astaxanthin used widely in aquaculture, pharmaceutical, and cosmetic industries. A new strain of Haematococcus (LCR‐26C‐1f) isolated from the New Zealand alpine zone was evaluated in this study. The influence of vitamins, micronutrients, various carbon and nitrogen sources were investigated to maximize biomass production in batch cultures using shake flasks. Supplementation of vitamins consisting of thiamine, biotin, and cyanocobalamin improved the cell density by 40% over the vitamin‐free medium. Out of the individual vitamins tested, thiamine was shown to be necessary to maintain high cell densities. The best nitrogen source tested was nitrate in the form of sodium nitrate_, at a 40 mM concentration. Heterotrophic growth yielded much lower cell densities compared to autotrophic growth. The micronutrients iron and manganese were essential for growth. However, the best growth was obtained using a micronutrient mix that included iron, copper, cobalt, zinc, manganese and molybdenum.     

IRON-MEDIATED CHANGES IN THE GROWTH OF LAKE ERIE PHYTOPLANKTON AND AXENIC ALGAL CULTURES1

The effect of iron enrichment on algal growth and photosynthesis was investigated using natural assemblages of Lake Erie phytoplankton and axenic cultures of Anabaena, Scenedesmus and Selenastrum. Cell yield and photosynthesis were frequently inhibited in the presence of unchelated iron over the range of 3.6 to 53.7 μM iron as FeCl₃. In lake water and in a defined medium with low nutrient concentrations, the degree of inhibition by iron could be reduced by chelating the iron with EDTA or by enriching the cultures with phosphorus. Chemical analyses revealed that the EDTA efectively reduced the ability of the ferric iron to remove soluble phosphorus from the media. EDTA was also observed to reduce rather than enhance iron uptake by axenic cultures of A. flos-aquae. These data support the hypothesis that additions of EDTA to low-nutrient media may serve to stimulate algal growth in the presence of iron by preventing the iron from altering extracellular concentrations of soluble ions essential for algal metabolism. In medium with high nutrient concentrations, the soluble phosphorus concentration was not appreciably altered by either EDTA-chelated or unchelated iron enrichment (0.9 to 53.7 μM). Instead, the observed enhancement of cell yield by EDTA-chelated iron in nutrient-rich media appeared to be due to the direct effect of iron on intracellular metabolic processes.     

Effect of pH, temperature and nutrient limitations on growth and leukotoxin production by Mannheimia haemolytica in batch and continuous culture

AIMS: Quantification of the effects of pH, temperature and nutrient limitations on the growth and leukotoxin (LKT) production parameters of Mannheimia haemolytica in batch and chemostat culture. METHODS AND RESULTS: Mannheimia haemolytica strains OVI-1 and PH12296 were grown aerobically in two semi-defined media. In amino acid-limited cultures, the LKT concentration and yield in terms of biomass (Y(LKT/x)) were up to eightfold greater than in carbon-limited cultures. Supplementing amino acid-limited chemostat cultures with cysteine, glutamine, ferric iron and manganese further enhanced the Y(LKT/x) values up to threefold. Supplementation of an amino acid-limited batch culture of M. haemolytica strain OVI-1 with these nutrients resulted in an LKT concentration of 1.77 g l(-1) that was 45-fold greater than that obtained in RPMI 1640 medium. Aerobiosis enhanced LKT production. High acetic acid concentrations were produced under carbon-sufficient conditions. The highest maximum specific growth rates were recorded in the range of pH 6.8 to 7.8 and 37 to 40 degrees C. CONCLUSIONS: An amino acid-limited culture medium greatly improved LKT production in aerobic batch culture, which could be further enhanced by supplementation with cysteine, glutamine, ferric iron and manganese. SIGNIFICANCE AND IMPACT OF THE STUDY: It was demonstrated that LKT production by M. haemolytica could be dramatically increased through manipulation of the culture medium composition, which could benefit the production of LKT-based vaccines against bovine shipping fever pneumonia.     

BACTERIAL INFLUENCE UPON GROWTH AND HYDROCARBON PRODUCTION OF THE GREEN ALGA BOTRYOCOCCUS BRAUNII1

Batch cultures of the hydrocarbon-rich alga Botryococcus braunii, Kütz. (axenic strains, non-axenic strains, associations with selected microorganisms) were examined with regard to total biomass and hydrocarbons at the onset of the stationary phase. Pronounced variations, related to the origin of the strains and to growth conditions, were observed with axenic cultures. It also appeared that the presence of microorganisms is not essential for high hydrocarbon production. Nevertheless, numerous bacteria were shown to exert considerable influence, antagonistic or beneficial, on B. braunii growth yield and hydrocarbon production. Such effects were strongly dependent on the species involved and on culture conditions. The presence of various microorganisms can influence not only the quantity of hydrocarbons produced, but also their level in the algal biomass and their relative abundance. However, their chemical structure is not affected. Intricate relationships were observed in B. braunii-bacteria systems and numerous factors (including, in some cultures, large positive effects due to bacterially produced CO₂) were implicated. Accordingly, specific associations should provide appropriate conditions for renewable hydrocarbon production via B. braunii large scale cultures.     

Variability of selected trace elements of different meat cuts determined by ICP-MS and DRC-ICPMS

The aim of this study was to determine the levels of cadmium, lead, iron, zinc, selenium, manganese, copper and molybdenum in different cuts of beef, pork, lamb, chicken and foal collected from supermarkets and butcheries in Switzerland. The concentrations of manganese, copper, molybdenum, zinc, iron, selenium, cadmium and lead were determined by inductively coupled plasma mass spectrometry (ICP-MS) after microwave digestion. Mean values and their respective coefficients of variation were calculated from the measured concentrations. The concentrations found for cadmium and lead ranged from 0.6 to 3.9 μg/100 g and 1.0 to 2.1 μg/100 g, respectively. Concentrations ranged between 0.5 and 3.3 mg/100 g for iron, 0.7 and 5.1 mg/100 g for zinc, 9 and 44 μg/100 g for selenium, 3.1 and 16.7 μg/100 g for manganese, 0.3 and 132 μg/100 g for copper and 0.9 and 3.2 μg/100 g for molybdenum. Differences found for the concentrations in meat from different species as well as between the individual meat cuts were notable for iron, zinc, selenium and copper. Manganese concentrations were found to vary unsystematically within muscles and species. Molybdenum concentrations were higher in chicken meat in comparison with the mammalian meats. The highest coefficients of variation were found for manganese (13% to 142%) and copper (13% to 224%), while the lowest was found for zinc (4% to 45%). In conclusion, in order to provide an accurate overview and to be able to calculate reliable dietary intakes, it is important to include the variability in food composition data.     

Intraspecific variability in growth response to cadmium of the wood-rotting fungus Piptoporus betulinus

The intraspecific variability in growth response to cadmium (Cd) on agar media and in liquid culture was studied among fourteen strains of a wood-rotting fungus Piptoporus betulinus. The variability of Cd tolerance was found to be very high. The ED(50) ranged from 6.8 μM Cd in the most sensitive strain, up to 255.1 μM in the most resistant one. On agar media the addition of Cd to nutrient media resulted in reduction of relative growth rate and increased lag time. While the reduction of growth rate was already apparent at 10 μM Cd, the lag time was significantly increased in higher Cd concentrations. Five strains of P. betulinus failed to grow at 250 μM Cd and none grew at 500 μM metal. Biomass production in liquid culture was less sensitive to addition of Cd than the growth rate on solid media. At 100 μM Cd the radial growth rate of the mycelium was reduced to 27%, whereas the dry mass of mycelium was 77% of the respective control value. A group of four Cd-sensitive strains was found, showing low metal tolerance both on solid media and in liquid cultures. Although the isolates originated from sites with different Cd-pollution level, no correlation between level of Cd-pollution and resistance (ED(50)) was found. The growth rate of fourteen tested strains displayed lower variability than biomass production, showing that radial growth rate is more species-specific and therefore more valuable for interspecific comparisons of growth response.     

TRACE-ELEMENT TOXICITIES IN OAT PLANTS

Excessive amounts of nickel, cobalt, chromium, copper, zinc, manganese, molybdenum and aluminium in nutrient solutions supplied to oat plants in sand culture produce ( a ) chlorosis and ( b ) other symptoms specific to the element involved. The specific symptoms are distinct for each metal, although those of cobalt and nickel might be confused.
The toxic effects of nickel, cobalt, copper, zinc, manganese and molybdenum are associated with high concentrations of the element in the leaf tissue, but this is not always so with chromium and aluminium.
The toxic effects of nickel, chromium, copper and molybdenum are associated with a reduced nitrogen content of the plant. Nickel, cobalt, chromium, zinc and manganese increase the concentration of phosphorus in the tissue whilst aluminium decreases it, probably to a deficiency level.
Aluminium reduces the intensity of toxic symptoms produced by nickel—probably by reducing the uptake of nickel and phosphorus. Copper effectively reduces the leaf necrosis produced by nickel, but not the nickel content of the leaf tissue; it is suggested that one factor in nickel toxicity may be inhibition of one or more functions of copper. The other elements slightly increase chlorosis and some increase necrosis.
The order of activitjl of the elements in producing chlorosis is found to be Ni>Cu>Co>Cr>Zn>Mo>Mn. This order, which is related to that giving yield reduction and is similar to the order of stability of metal complexes, is discussed in relation to induced iron deficiency.     

The impact of petroleum hydrocarbons (diesel) on periphyton in an impacted tropical estuary based on in situ microcosms

The distribution of petroleum hydrocarbons and their effects on the periphytic algal biomass using in situ microcosms were investigated in Ponggol estuary located on the northeastern coast of Singapore. Dissolved or dispersed petroleum hydrocarbon (DDPH) concentrations in the surface and bottom waters and absorbed or adsorbed petroleum hydrocarbon (AAPH) concentrations in sediments were monitored from July 1999 to June 2000. Results showed concentrations ranging from 4.42 to 248.94 μg l⁻¹, from 0.35 to 1099.65 μg l⁻¹, and from 20.55 to 541.01 mg kg⁻¹ for DDPH in surface and bottom waters and AAPH in sediments, respectively. Accidental spillages of fuel from dredgers operating in the estuary, fuel and engine oil from recreational boats, shipping operations in the adjacent strait, and runoff monsoon drains in the vicinity were some of the possible sources of petroleum hydrocarbons in the estuary. An assessment of environmentally realistic concentrations of petroleum hydrocarbons on periphytic algal biomass using in situ microcosms revealed signs of acute toxicity. A reduction in periphytic algal biomass (with respect to controls) of 68-93% was observed for various treatments exposed to diesel.     

LOSS OF NUTRIENTS FROM WATER SAMPLES BY FILTRATION AND ITS AFFECT ON ALGAL BIOASSAY PROCEDURES

SUMMARY

Water samples were collected from fourteen sampling points along the Hunyani River system and subjected to various filtration treatments involving glass fibre filters and 1,2 and 0,45 pm membrane filters. Chemical analyses of the filtered waters showed that nitrogen, phosphorus and iron were lost by different filtration treatments. Filtration by membrane filters led to a reduction of algal growth potential as demonstrated by algal bioassays using Selanastrwn capricornutum Printz as the test alga.

Algal bioassays showed that biologically available phosphorus was primarily removed by the 1,2 μm membrane filter while biologically available nitrogen and iron was principally lost by filtration through the 0,25 μm membrane filter. A refined algal bioassay designed to determine the identity of limiting micronutrients found that all essential micronutrients were affected by membrane filtration.     

Effect of iron on growth and lipid accumulation in Chlorella vulgaris

The economic feasibility of algal mass culture for biodiesel production is enhanced by the increase in biomass productivity and storage lipids. Effect of iron on growth and lipid accumulation in marine microalgae Chlorella vulgaris were investigated. In experiment I, supplementing the growth media with chelated FeCl3 in the late growth phase increased the final cell density but did not induce lipid accumulation in cells. In experiment II, cells in the late-exponential growth phase were collected by centrifugation and re-inoculated into new media supplemented with five levels of Fe3+ concentration. Total lipid content in cultures supplemented with 1.2 x 10(-5) mol L(-1) FeCl3 was up to 56.6% biomass by dry weight and was 3-7-fold that in other media supplemented with lower iron concentration. Moreover, a simple and rapid method determining the lipid accumulation in C. vulgaris with spectrofluorimetry was developed.     

Response surface methodology as a decision-making tool for optimization of culture conditions of green microalgae Chlorella spp. for biodiesel production

We have evaluated process optimization and the interactive effects of a number of variables using a Box–Behnken design of response surface methodology (RSM). The process variables nitrate, phosphate, glucose and pH were optimized to enhance the cell growth rate, lipid accumulation and other biochemical parameters of Chlorella spp. The most significant increase in lipid production (dry cell weight basis) occurred at limited concentrations of nitrate and phosphate, 1 % glucose and pH 7.5. The addition of nitrates during the mid-lag and mid-exponential phases produced the maximum inhibitory effect on lipid accumulation and the presence of yeast extract led to a further enhancement of lipid accumulation. Of all the media tested, BG-11 was the best suited medium for algal biomass production and chlorophyll content. A significant increase in algal biomass was observed in BG-11 supplemented with bicarbonate and glucose (1 %). The maximum specific growth rate observed was on 9th day of culturing. Results of optimization of process variables through response surface methodology and optimization of various other conditions reflect cutting edge research directed towards increasing algal biomass and lipid content for biodiesel production using an efficient economical technological approach.     

Trace element uptake by L-cells as a function of trace elements in a synthetic growth medium

The concentration of trace elements in L-cells has been studied as a function of the trace metal content of the growth medium. Cells were cultured in synthetic media which contained varying trace amounts of the elements manganese, iron, cobalt, copper, zinc and molybdenum. The cellular concentration of the elements potassium, iron, copper and zinc were then determined. It was found that the cell accumulates trace metals at a different rate than they are made available. Deficiencies in zinc could be “induced” in the cell by increasing the concentration of iron, manganese and cobalt; cellular iron deficiencies were observed at larger medium concentrations of zinc, manganese, copper and cobalt. Trace metal uptake by the cell was seen to parallel the utilization by multicellular organisms.     

Effect of metal ions on aflatoxin production byAspergillus parasiticus

The effect of iron, copper, cobalt, cadmium, zinc, molybdenum, magnesium and manganese salts was studied on aflatoxin production in relation to mycelial mass. Iron, copper and cadmium salts decreased the aflatoxin production to different levels but a mixed trend was observed depending on salt concentration, with molybdenum, magnesium and manganese. Cobalt and zinc salts stimulated aflatoxin production at all concentrations studied. The maximum increase in aflatoxin production, 655% and 519% was observed in the presence of zinc sulfate and sodium molybdate, respectively. A negative correlation was observed between aflatoxin production and vegetative growth of fungus.     

Inhibitory effect of molybdenum and vanadium salts on aflatoxin B1 synthesis by Aspergillus flavus

The effects of the elements zinc, manganese, iron, copper, molybdenum, and vanadium, added in various salt forms, on mycelial weights and aflatoxin B1 accumulation in the mycelium of Aspergillus flavus were investigated in liquid shake cultures. Ammonium heptamolybdate, when added to a complete medium at concentrations of 50-100 mg/L, appreciably reduced aflatoxin B1 accumulation without affecting growth of the fungus. Sodium molybdate and sodium monovanadate also reduced aflatoxin B1 yields without affecting mycelial growth but to a lesser extent. The addition of zinc sulphate stimulated aflatoxin B1 production in all media used. The influence of the other trace elements on aflatoxin production depended on the level of trace elements present in the basal medium. In general, manganese chloride had a stimulatory effect, whereas copper sulphate depressed yields. Mycelial levels of aflatoxin had peaked and then declined before mycelial dry weights had reached maximum. High yields of aflatoxin B1 were obtained in media having a final pH as low as pH 2.8.     

Studies on batch and continuous cultures of Botryococcus braunii: hydrocarbon production in relation to physiological state, cell ultrastructure, and phosphate nutrition

The growth of the hydrocarbon-rich alga Botryococcus braunii was studied under "air-lift" conditions using batch and continuous cultures. Large variations in the physiological state of B. braunii were achieved in batch cultures and in continuous cultures with various dilution rates. The possible effects of these variations upon hydrocarbons (nature, relative abundance, location, level, productivity) and also on the production of exocellular polysaccharides were examined. The relationships between the physiological state of B. braunii and its hydrocarbon and polysaccharide production were discussed and compared with those generally observed in unicellular algae. The factors giving rise to the transition from high to low productivity stages were considered. To this end we examined, at first, the variations in cell ultrastructure and the resulting degeneration occurring during batch cultures. Afterward the parallel changes in some parameters of the medium (pH, phosphate level) were determined and their possible relationships with B. braunii growth and hydrocarbon production were discussed. The main features of phosphate nutrition in B. braunii and its effects on hydrocarbons were finally examined.