Effects of alkalinity and co-substrate on the performance of an upflow anaerobic sludge blanket (UASB) reactor through decolorization of Congo Red azo dye

The effect of substrate (glucose) concentrations and alkalinitiy (NaHCO3) on the decolorization of a synthetic wastewater containing Congo Red (CR) azo dye was performed in an upflow anaerobic sludge blanket (UASB). Color removal efficiencies approaching 100% were obtained at glucose-COD concentrations varying between 0 and 3000 mg/l. The methane production rate and total aromatic amine (TAA) removal efficiencies were found to be 120 ml per day and 43%, respectively, while the color was completely removed during glucose-COD free operation of the UASB reactor. The complete decolorization of CR dye under co-substrate free operation could be attributed to TAA metabolism which may provide the electrons required for the cleavage of azo bond in CR dye exist in the UASB reactor. No significant differences in pH levels (6.6-7.4), methane production rates (2000-2700 ml/day) and COD removal efficiencies (82-90%) were obtained for NAHCO3 concentrations ranging between 550 and 3000 mg/l. However, decolorization efficiency remained at 100% with decreasing NaHCO3 concentrations as low as 250 mg/l in the feed. An alkalinity/COD ratio of 0.163 in the feed was suggested for simultaneous optimum COD and color removal.     

Septum-localized protein required for filament integrity and diazotrophy in the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120

Heterocysts, formed when filamentous cyanobacteria, such as Anabaena sp. strain PCC 7120, are grown in the absence of combined nitrogen, are cells that are specialized in fixing atmospheric nitrogen (N(2)) under oxic conditions and that transfer fixed nitrogen to the vegetative cells of the filament. Anabaena sp. mutants whose sepJ gene (open reading frame alr2338 of the Anabaena sp. genome) was affected showed filament fragmentation and arrested heterocyst differentiation at an early stage. In a sepJ insertional mutant, a layer similar to a heterocyst polysaccharide layer was formed, but the heterocyst-specific glycolipids were not synthesized. The sepJ mutant did not exhibit nitrogenase activity even when assayed under anoxic conditions. In contrast to proheterocysts produced in the wild type, those produced in the sepJ mutant still divided. SepJ is a multidomain protein whose N-terminal region is predicted to be periplasmic and whose C-terminal domain resembles an export permease. Using a green fluorescent protein translationally fused to the carboxyl terminus of SepJ, we observed that in mature heterocysts and vegetative cells, the protein is localized at the intercellular septa, and when cell division starts, it is localized in a ring whose position is similar to that of a Z ring. SepJ is a novel composite protein needed for filament integrity, proper heterocyst development, and diazotrophic growth.     

Synthesis of nitrogenase and heterocysts by Anabaena sp. CA in the presence of high levels of ammonia.

Anabaena sp. CA fails to synthesize heterocysts and nitrogenase when grown with KNO3 as the nitrogen source. By contrast, both heterocysts and proheterocysts are synthesized in NH4Cl-containing media to a level nearly commensurate with cells grown in the absence of combined nitrogen. The growth rate of the organism in NH4Cl-containing media was similar to that obtained with KNO3 as the nitrogen source and was independent of the presence of N2 in the atmosphere. Thus, our results indicate that the organism assimilated nitrate and ammonium nitrogen equally well to meet the nitrogen requirements for growth. Moreover, in contrast to previous studies with other cyanobacteria, the repressor singal for heterocyst differentiation in Anabaena sp. CA is not derived from the metabolism of ammonia but appears to be involved with nitrate metabolism. Nitrogenase activity was partially expressed in NH4Cl-grown cultures. Increasing the level of nitrogenase activity to a value representative of a N2-grown culture required both the inhibition of ammonia assimilation and de novo protein synthesis. An increase in the number of mature heterocysts was not required. The fact that high levels of exogenous ammonia only partially repress the synthesis of proteins required for the maximum expression of nitrogenase activity in Anabaena sp. CA has important implications.     

Chemoheterotrophic Growth of the Cyanobacterium Anabaena sp. Strain PCC 7120 Dependent on a Functional Cytochrome c Oxidase

Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium commonly used as a model organism for studying cyanobacterial cell differentiation and nitrogen fixation. For many decades, this cyanobacterium was considered an obligate photo-lithoautotroph. We now discovered that this strain is also capable of mixotrophic, photo-organoheterotrophic, and chemo-organoheterotrophic growth if high concentrations of fructose (at least 50 mM and up to 200 mM) are supplied. Glucose, a substrate used by some facultatively organoheterotrophic cyanobacteria, is not effective in Anabaena sp. PCC 7120. The gtr gene from Synechocystis sp. PCC 6803 encoding a glucose carrier was introduced into Anabaena sp. PCC 7120. Surprisingly, the new strain containing the gtr gene did not grow on glucose but was very sensitive to glucose, with a 5 mM concentration being lethal, whereas the wild-type strain tolerated 200 mM glucose. The Anabaena sp. PCC 7120 strain containing gtr can grow mixotrophically and photo-organoheterotrophically, but not chemo-organoheterotrophically with fructose. Anabaena sp. PCC 7120 contains five respiratory chains ending in five different respiratory terminal oxidases. One of these enzymes is a mitochondrial-type cytochrome c oxidase. As in almost all cyanobacteria, this enzyme is encoded by three adjacent genes called coxBAC1. When this locus was disrupted, the cells lost the capability for chemo-organoheterotrophic growth.     

Isolation and characterization of mutants of two diazotrophic cyanobacteria tolerant to high concentrations of inorganic carbon

Diazotrophic heterocystous cyanobacteria Nostoc calcicola and Anabaena sp. ARM 629 were investigated for their ability to grow in presence of sodium bicarbonate (NaHCO3) or carbon dioxide (CO2) under cultural conditions. Maximum growth was observed in 75 mM NaHCO3 and 5% CO2 in N. calcicola and Anabaena ARM 629, respectively. Although their growth rate declined, N. calcicola and Anabaena sp. could tolerate upto 250 mM NaHCO3 and 20% CO2, respectively. N-methyl-N'-nitro N nitrosoguanidine induced mutants of these cyanobacteria were isolated which showed growth upto 1 M NaHCO3 (N. calcicola) or 50% CO2 (Anabaena sp.) in comparison to their wild types. The mutants also showed cross-resistance to either of the inorganic carbon compounds, which was not observed for wild type. It was concluded that mutants were altered in multiple properties enabling them to grow at elevated levels of inorganic carbon compounds.     

Cyanobacteria in Uruguayan rice fields: diversity, nitrogen fixing ability and tolerance to herbicides and combined nitrogen

It has been established that cyanobacteria play a vital role in the maintenance of flooded rice field fertility. To evaluate the potential use of nitrogen-fixing cyanobacteria as a natural biofertilizer for rice in Uruguay, the diversity, abundance and nitrogen fixing ability of these microorganisms were studied in the field and in the laboratory. The effect of urea fertilization on population density and diversity of heterocystous cyanobacteria was determined on a 3-year assay. The highest number of cyanobacteria, 1.6x10(4) CFU x m(-2), was found at the control 8 weeks after flooding. About 90% of the heterocystous cyanobacteria found in both treatments belong to the genera Nostoc and Anabaena. Maximal nitrogenase activity was reached after 12 weeks of flooding in both treatments, with an average of about 20 micromol C2H4 x m(-2) x h(-1). To improve the understanding of the environmental factors that can limit nitrogenase activity in rice fields, two of the most abundant cyanobacteria isolates were tested for tolerance to combined nitrogen and two herbicides. In both isolates 0.2 mM ammonium inhibited nitrogenase activity after 24 h of culture. The addition of field-recommended doses of quinclorac and propanil affected oxygen photoevolution but nitrogenase activity was only inhibited by propanil.     

Aerobic degradation of the azo dye acid red 151 in a sequencing batch biofilter

The azo dye acid red 151 (AR151) was aerobically biodegraded in a sequencing batch biofilter packed with a porous volcanic rock. AR151 was used as the sole source of carbon and energy for acclimated microorganisms. Acclimation was followed using the degradation time and the oxygen uptake rate. A maximal oxygen uptake rate of 0.5 mg O(2)/(lmin) was obtained. Mineralization studies showed that 73% (as carbon) of the initial azo dye was transformed to CO(2) by the consortia. A maximal substrate degradation rate of 247 mg AR151/(l(reactor)d) was obtained. Color removal was up to 99% using an initial concentration of 50 mg AR151/l. Anaerobic tests suggested that in the interior of the porous material, anaerobic biotransformations can occur, contributing from 14% to 16% of the decoloration of the azo dye.     

Decolourization of azo dyes and a dye industry effluent by a white rot fungus Thelephora sp

A white rot fungus Thelephora sp. was used for decolourization of azo dyes such as orange G (50 microM), congo red (50 microM), and amido black 10B (25 microM). Decolourization using the fungus was 33.3%, 97.1% and 98.8% for orange G, congo red and amido black 10B, respectively. An enzymatic dye decolourization study showed that a maximum of 19% orange G was removed by laccase at 15 U/ml whereas lignin peroxidase (LiP) and manganese dependent peroxidase (MnP) at the same concentration decolourized 13.5% and 10.8%, orange G, respectively. A maximum decolourization of 12.0% and 15.0% for congo red and amido black 10B, respectively, was recorded by laccase. A dye industry effluent was treated by the fungus in batch and continuous modes. A maximum decolourization of 61% was achieved on the third day in the batch mode and a maximum decolourization of 50% was obtained by the seventh day in the continuous mode. These results suggest that the batch mode of treatment using Thelephora sp. may be more effective than the continuous mode for colour removal from dye industry effluents.     

Salicylate biodegradation by various algal-bacterial consortia under photosynthetic oxygenation

Four green microalgae (Chlorella sorokiniana, Chlorella vulgaris, Scenedesmus obliquus and Selenastrum capricornutum), a wild Bolivian microalga strain and two cyanobacteria (Anabaena catenula and Microcystis aeruginosa) were compared for tolerance to salicylate, O2 production capacity and ability to support salicylate degradation by a Ralstonia basilensis strain in symbiotic microcosms with the microalgae. Microcystis aeruginosa had the highest tolerance to salicylate at 500 mg l(-1) and 1500 mg l(-1) but only produced 0.7 mg O2 l(-1) h(-1) in the absence of pollutant. Chlorella sorokiniana resisted salicylate at 1500 mg l(-1) with the highest O2 production in the absence of salicylate (26 mg l(-1) h(-1)) closely followed by the Bolivian microalga (23 mg l(-1) h(-1)) and Chlorella vulgaris (21 mg l(-1) h(-1)). Selenastrum capricornutum and Anabaena catenula were completely inhibited by salicylate at 500 mg l(-1). When inoculated with Ralstonia sp. and supplied with salicylate, Chlorella sorokiniana had the highest removal rate (19 mg l(-1) h(-1)), followed by the wild Bolivian strain (18 mg l(-1) h(-1)) and Chlorella vulgaris (14 mg l(-1) h(-1)).     

Use of recombinant aequorin to study calcium homeostasis and monitor calcium transients in response to heat and cold shock in cyanobacteria

We investigated the possibility of Ca(2+) signaling in cyanobacteria (blue-green algae) by measuring intracellular free Ca(2+) levels ([Ca(2+)](i)) in a recombinant strain of the nitrogen fixing cyanobacterium Anabaena strain sp. PCC7120, which constitutively expresses the Ca(2+)-binding photoprotein apoaequorin. The homeostasis of intracellular Ca(2+) in response to increasing external Ca(2+) has been studied in this strain. The resting level of free Ca(2+) in Anabaena was found to be between 100 and 200 nM. Additions of increasing concentrations of external Ca(2+) gave a transient burst of [Ca(2+)](i) followed by a very quick decline, reaching a plateau within seconds that brought the level of [Ca(2+)](i) back to the resting value. These results indicate that Anabaena strain sp. PCC7120 is able to regulate its internal Ca(2+) levels. We also monitored Ca(2+) transients in our recombinant strain in response to heat and cold shock. The cell's response to both stresses was dependent on the way they were induced. The use of inhibitors suggests that heat shock mobilizes cytosolic Ca(2+) from both intracellular and extracellular sources, while the Ca(2+) source for cold shock signaling is mostly extracellular.     

EFFECT OF SODIUM AND NITRATE ON GROWTH OF ANABAENA FLOS-AQUAE (CYANOPHYTA)1

The growth response of a nitrogen fixing cyanobacterium, Anabaena flos-aquae (Lyng) Bréb., to sodium and nitrate was examined in batch culture under controlled laboratory conditions. Sodium (range 0-12 mg Na⁺· L^-1) enhanced growth of the cyanobacterium under nitrate-sufficient (5.7 mg NO³-N · L^-1) but not nitrate-limited (0.49 mg NO³- N · L^-1) conditions. The magnitude of the growth response was related to the nutritional history of the culture. No significant effect of sodium on nitrate utilization was observed. The increase in ambient sodium levels in many lakes may provide a competitive advantage to cyanobacteria.     

Degradation of azo dyes containing aminonaphthol by Sphingomonas sp strain 1CX

Sphingomonas sp strain 1CX was isolated from a wastewater treatment plant and is capable of aerobically degrading a suite of azo dyes, using them as a sole source of carbon and nitrogen. All azo dyes known to be decolorized by strain 1CX (Orange II, Acid Orange 8, Acid Orange 10, Acid Red 4, and Acid Red 88) have in their structure either 1-amino-2-naphthol or 2-amino-1-naphthol. In addition, an analysis of the structures of the dyes degraded suggests that there are certain positions and types of substituents on the azo dye which determine if degradation will occur. Growth and dye decolorization occurs only aerobically and does not occur under fermentative or denitrification conditions. The mechanism by which 1CX decolorizes azo dyes appears to be through reductive cleavage of the azo bond. In the case of Orange II, the initial degradation products were sulfanilic acid and 1-amino-2-naphthol. Sulfanilic acid, however, was not used by 1CX as a growth substrate. The addition of glucose or inorganic nitrogen inhibited growth and decoloration of azo dyes by 1CX. Attempts to grow the organism on chemically defined media containing several different amino acids and sugars as sources of nitrogen and carbon were not successful. Phylogenetic analysis of Sphingomonas sp strain 1CX shows it to be related to, but distinct from, other azo dye-decolorizing Sphingomonas spp strains isolated previously from the same wastewater treatment facility. Received 19 May 1999/ Accepted in revised form 11 August 1999     

Isolation and characterization of transposon-induced chlorate resistant mutants of the cyanobacterium Anabaena species PCC 7120

Mutants of Anabaena sp. PCC 7120 resistant to chlorate were isolated using transposon mutagenesis. The Anabaena population of 5 x 10(7) cells ml(-1) and log phase Escherichia coli cultures in undisturbed conditions produced maximum exconjugants. Nitrate-promoted growth and cellular constituents observed in the parent were absent in the mutants. Nitrate repressed heterocyst formation and N2-fixation in the parent, but had little or no effect on the mutants.     

六种固氮蓝藻提取液对玉米的促长作用和提取液成分比较

本文用六种固氮蓝藻的提取液处理玉米种子,同时对其提取液氨基酸组成和碳水化合物与维生素Ｂ１２的含量进行了分析。结果表明固氮鱼腥藻ＨＢ６８６（ＡｎａｂａｅｎａａｚｏｔｉｃａＨＢ６８６）、球孢鱼腥藻ＨＢ１０１７（Ａ．ｓｐｈａｅｒｉｃａＨＢ１０１７）、多变鱼腥藻ＨＢ１０５８（Ａ．ｖａｒｉａｂｉｌｉｓＨＢ１０５８）和小单歧藻ＨＢＴＴ（ＴｏｌｙｐｏｔｈｒｉｘｔｅｎｕｉｓＨＢＴＴ）的提取液中氨基酸、碳水化合物和维生素Ｂ１２的含量高于鱼腥藻ＳＰ．ＨＢ１０４２（Ａｎａｂａｅｎａｓｐ．ＨＢ１０４２）和繁育管链藻ＨＢ３８（ＡｕｌｏｓｉｒａｆｅｒｔｉｌｉｓｓｉｍａＨＢ３８）。同时,促进玉米种子萌发和幼苗生长的效果前四种藻较好,后两种则较差。     

Decolorization of azo dyes by Shewanella sp. under saline conditions

Wastewaters from textile processing and dye-stuff manufacture industries contain substantial amounts of salts in addition to azo dye residues. To examine salinity effects on dye-degrading bacteria, a study was carried out with four azo dyes in the presence of varying concentrations of NaCl (0-100 g l(-1)) with a previously isolated bacterium, Shewanella putrefaciens strain AS96. Under static, low oxygen conditions, the bacterium decolorized 100 mg dye l(-1) at salt concentrations up to 60 g NaCl l(-1). There was an inverse relationship between the velocity of the decolorization reaction and salt concentration over the range between 5 and 60 g NaCl l(-1) and at dye concentrations between 100 and 500 mg l(-1). The addition of either glucose (C source) or NH(4)NO(3) (N source) to the medium strongly inhibited the decolorization process, while yeast extract (4 g l(-1)) and Ca(H(2)PO(4))(2).H(2)O (1 g l(-1)) both enhanced decolorization rates. High-performance liquid chromatography analysis demonstrated the presence of 1-amino-2-naphthol, sulfanilic acid and nitroaniline as the major metabolic products of the azo dyes, which could be further degraded by a shift to aerobic conditions. These findings show that Shewanella could be effective for the treatment of dye-containing industrial effluents containing high concentrations of salt.     

Regulation of nitrate reductase cellular levels in the cyanobacteria Anabaena variabilis and Synechocystis sp.

Abstract The effect of the nitrogen source on the cellular activity level of assimilatory nitrate reductase in the cyanobacteria Anabaena variabilis (ATCC29413) and Synechocystis sp. (PCC6714) has been examined. In the filamentous N₂-fixing A. variabilis , nitrate behaved as a nutritional inducer of nitrate reductase, with ammonium acting (via products of its assimilation) as an antagonist with regard to nitrate. Ammonium-promoted repression of nitrate reductase was also evident in the unicellular non-nitrogen fixer Synechocystis , but in this strain nitrate was not required as an obligatory inducer.     

Pesticide (hexachlorocyclohexane) inhibition of growth and nitrogen fixation in blue-green algae Anabaenopsis raciborskii and Anabaena aphanizomenoides.

The effects of the pesticide hexachlorocyclohexane (HCH) on the nitrogen fixing blue-green algae Anabaenopsis raciborskii and Anabaena aphanizomenoides commonly found as blooms in fish ponds were studied. These algae were very sensitive to HCH, and a distinct decrease in growth rate was observed on prolonged incubation. Lower concentrations (10 microgram/ml) were algistatic and higher concentrations (60 microgram/ml) were algicidal. The inhibition of nitrogen fixation indicated that the presence of HCH might affect overall nitrogen economy of inland waters.     

Hydrogen formation by cyanobacteria cultures selected for nitrogen fixation

Seventy-one cyanobacteria containing cultures were enriched from various soil and water locations either under aerobic and/or anaerobic conditions on agar medium selective for nitrogen fixation. Kept under argon containing 1% CO₂ for 24 and 48 h most of these cultures evolved hydrogen at very variable rates up to 116 l per mg chlorophyll and hour as a mean value over a time period of 24h. Several samples evolved hydrogen more efficiently compared with known hydrogen producing pure strains from culture collections. Thirty-one of the investigated cultures showed a hydrogen formation higher than 10 l per mg chlorophyll and hour measured over 24 or 48 h. Among these all the morphological forms of cyanobacteria i.e. unicellular and filamentous with or without heterocysts are found. Hence, selecting for nitrogen fixing cyanobacteria seems to be a practical method to find efficient hydrogen producers.     

鱼腥藻PCC 7120乙酰辅酶A合成酶All0769的缺失降低异形胞频率

研究鉴定了All0769为鱼腥藻PCC 7120中乙酰辅酶A合成酶,通过CRISPR/Cpf1系统敲除鱼腥藻PCC7120中的乙酰辅酶A合成酶(由all0769编码),探究了乙酰辅酶A合成酶在异形胞分化中的调控机制。结果所示:All0769能在体外反应中催化乙酰辅酶A的生成。在供氮环境下,敲除all0769会影响藻细胞生长速率。而无论环境中是否存在化合氮,Δall0769突变株的乙酰辅酶A和α-酮戊二酸含量均显著减少。在供氮环境下,Δall0769突变株中检测到(26.17±1.55) nmol/mg protein的乙酰辅酶A,而在野生型中检测出(43.04±1.09) nmol/mg的乙酰辅酶A。Δall0769突变株的α-酮戊二酸[(1.41±0.24) nmol/mg protein]低于野生型的α-酮戊二酸[(2.13±0.05) nmol/mg protein]。在缺氮环境下,Δall0769突变株中检测到(10.00±2.81) nmol/mg protein的乙酰辅酶A,而在野生型中检测出(29.82±4.04) nmol/mg protein的乙酰辅酶A。Δall07...