Microbiological and chemical characteristics of beaches along the Taranto Gulf (Ionian Sea, Southern Italy).

Coastal habitats provide important ecosystem services, such as the maintenance of ecological sustainability, water quality regulation, nutrient recycling, and sandy beaches which are important areas for recreation and tourism. The quality of seawater is generally measured by determining the concentrations of Escherichia coli and intestinal Enterococci, which might be affected by the persistent populations of these bacteria in sand. Sand might thus be a significant source of pathogen exposure to beachgoers. The quality of coastal recreational waters can also be affected by eutrophication, water discoloration, and harmful algal blooms, which pose additional human health risks. Here, we conducted a monitoring of the beaches quality along the Taranto Gulf by determining the concentrations of fecal indicator organisms, as well as other parameters that are not traditionally measured (physicochemical parameters, Pseudomonas aeruginosa, and harmful microalgae), in shallow seawater and sand sampled from three beaches. The concentrations of bacteria were determined using both standard microbiological methods and the IDEXX system. Our results demonstrate the utility of measuring a greater number of parameters in addition to those conventionally measured, as well as the importance of assessing the health risks posed by the sand matrix. Additional work is needed to develop rapid analytical techniques that could be used to monitor the microbiological parameters of solid matrices.

Confocal hyperspectral microscopic imager for the detection and classification of individual microalgae.

We propose a confocal hyperspectral microscopic imager (CHMI) that can measure both transmission and fluorescent spectra of individual microalgae, as well as obtain classical transmission images and corresponding fluorescent hyperspectral images with a high signal-to-noise ratio. Thus, the system can realize precise identification, classification, and location of microalgae in a free or symbiosis state. The CHMI works in a staring state, with two imaging modes, a confocal fluorescence hyperspectral imaging (CFHI) mode and a transmission hyperspectral imaging (THI) mode. The imaging modes share the main light path, and thus obtained fluorescence and transmission hyperspectral images have point-to-point correspondence. In the CFHI mode, a confocal technology to eliminate image blurring caused by interference of axial points is included. The CHMI has excellent performance with spectral and spatial resolutions of 3 nm and 2 µm, respectively (using a 10× microscope objective magnification). To demonstrate the capacity and versatility of the CHMI, we report on demonstration experiments on four species of microalgae in free form as well as three species of jellyfish with symbiotic microalgae. In the microalgae species classification experiments, transmission and fluorescence spectra collected by the CHMI were preprocessed using principal component analysis (PCA), and a support vector machine (SVM) model or deep learning was then used for classification. The accuracy of the SVM model and deep learning method to distinguish one species of individual microalgae from another was found to be 96.25% and 98.34%, respectively. Also, the ability of the CHMI to analyze the concentration, species, and distribution differences of symbiotic microalgae in symbionts is furthermore demonstrated.

Cyanobacteria and microalgae growing on monuments of UNESCO World Heritage site Champaner Pavagadh, India: biofilms and their exopolysaccharide composition.

The present study investigated the biofilm organisms growing on selected monuments of the Champaner Pavagadh complex (Gujarat, India), which is a UNESCO World Heritage Site. The cyanobacteria and microalgae were isolated from biofilms collected through non-destructive methods. The identification of these biological organisms was done using micro-morphological characters and confirmed by 16S rRNA gene sequencing. The exopolysaccharide of each of the isolated strains was extracted, hydrolysed and analysed by the HPTLC. Six isolated strains representing five cyanobacteria and one microalga belong to the genera Desmonostoc, Nostoc, Leptolyngbya, Chroococcidiopsis and Asterarcys. The relationships between substrates' specificity of these isolated biofilm organisms and those identified globally were evaluated using maximum parsimony analysis to generate a consensus phylogenetic tree. The five strains of cyanobacteria isolated were closely clustered with cyanobacteria belonging to a tropical region. At the generic level, no relationship between the species and substratum specificity was recorded. The exopolysaccharide analysis of the isolated strains revealed the presence of seven monosaccharides. While glucose was present in all the analysed species, the concentration of either fucose or arabinose was high. The current study presents a novel HPTLC-based method for determination of monosaccharides composition from the extracellular polymeric substances.

Molecular and Structural Characterizations of Lipases from Chlorella by Functional Genomics.

Microalgae have been poorly investigated for new-lipolytic enzymes of biotechnological interest. In silico study combining analysis of sequences homologies and bioinformatic tools allowed the identification and preliminary characterization of 14 putative lipases expressed by Chlorella vulagaris. These proteins have different molecular weights, subcellular localizations, low instability index range and at least 40% of sequence identity with other microalgal lipases. Sequence comparison indicated that the catalytic triad corresponded to residues Ser, Asp and His, with the nucleophilic residue Ser positioned within the consensus GXSXG pentapeptide. 3D models were generated using different approaches and templates and demonstrated that these putative enzymes share a similar core with common α/ß hydrolases fold belonging to family 3 lipases and class GX. Six lipases were predicted to have a transmembrane domain and a lysosomal acid lipase was identified. A similar mammalian enzyme plays an important role in breaking down cholesteryl esters and triglycerides and its deficiency causes serious digestive problems in human. More structural insight would provide important information on the enzyme characteristics.

Potential of Exopolysaccharide from Porphyridium marinum to Contend with Bacterial Proliferation, Biofilm Formation, and Breast Cancer.

Exopolysaccharide (EPS) from marine microalgae are promising sources of a new generation of drugs. However, lot of them remain to be discovered and tested. In this study, EPS produced by Porphyridium marinum and its oligomers prepared by High Pressure Homogenizer have been tested for different biological activities, i.e., antibacterial, anti-fungal and antibiofilm activities on Candida albicans, as well as for their effects on the viability of murine breast cancer cells. Results have shown that all EPS samples present some biological activity. For antibacterial and antibiofilm activities, the native EPS exhibited a better efficiency with Minimum Inhibitory Concentration (MIC) from 62.5 µg/mL to 1000 µg/mL depending on the bacterial strain. For Candida albicans, the biofilm formation was reduced by about 90% by using only a 31.3 µg/mL concentration. Concerning breast cancer cells, lower molar masses fractions appeared to be more efficient, with a reduction of viability of up to 55%. Finally, analyses of polymers composition and viscosity measurements were conducted on all samples, in order to propose hypotheses involving the activities caused by the intrinsic properties of polymers.

Improvement of size-based particle separation throughput in slanted spiral microchannel by modifying outlet geometry.

The inertial microfluidic technique, as a powerful new tool for accurate cell/particle separation based on the hydrodynamic phenomenon, has drawn considerable interest in recent years. Despite numerous microfluidic techniques of particle separation, there are few articles in the literature on separation techniques addressing external outlet geometry to increase the throughput efficiency and purity. In this work, we report on a spiral inertial microfluidic device with high efficiency (>98%). Herein, we demonstrate how changing the outlet geometry can improve the particle separation throughput. We present a complete separation of 4 and 6 µm from 10 µm particles potentially applicable to separate microalgae (Tetraselmis suecica from Phaeodactylum tricornutum). Two spiral microchannels with the same cross section dimension but different outlet geometry were considered and tested to investigate the particle focusing behavior and separation efficiency. As compared with particle focusing observed in channels with a simple outlet, the particle focusing in a modified outlet geometry appears in a more successful focusing manner with complete separation. This simple approach of particle separation makes it attractive for lab-on-a-chip devices for continuous extraction and filtration of a wide range of cell/particle sizes.

Low-cost multi-core inertial microfluidic centrifuge for high-throughput cell concentration.

We developed a low-cost multi-core inertial microfluidic centrifuge (IM-centrifuge) to achieve a continuous-flow cell/particle concentration at a throughput of up to 20 mL/min. To lower the cost of our IM-centrifuge, we clamped a disposable multilayer film-based inertial microfluidic (MFIM) chip with two reusable plastic housings. The key MFIM chip was fabricated in low-cost materials by stacking different polymer-film channel layers and double-sided tape. To increase processing throughput, multiplexing spiral inertial microfluidic channels were integrated within an all-in-one MFIM chip, and a novel sample distribution strategy was employed to equally distribute the sample into each channel layer. Then, we characterized the focusing performance in the MFIM chip over a wide flow-rate range. The experimental results showed that our IM-centrifuge was able to focus various-sized particles/cells to achieve volume reduction. The sample distribution strategy also effectively ensured identical focusing and concentration performances in different cores. Finally, our IM-centrifuge was successfully applied to concentrate microalgae cells with irregular shapes and highly polydisperse sizes. Thus, our IM-centrifuge holds the potential to be employed as a low-cost, high-throughput centrifuge for disposable use in low-resource settings.

Photosynthetic Sponge-associated Eukaryotes in the Aegean Sea: A Culture-dependent Approach.

Symbioses between sponges and photosynthetic organisms are very diverse regarding the taxonomy and biogeography of both hosts and symbionts; to date, most research has focused on the exploration of bacterial diversity. The present study aims to characterize the culturable diversity of photosynthetic eukaryotes associated with sponges in the Aegean Sea, on which no information exists. Five microalgae strains were isolated from marine sponges; the strains were characterized by morphological features, and the 18S rRNA, 18S-28S Internal Transcribed Spacer, and ribulose-bisphosphate carboxylase large chain (rbcL) sequences. Our polyphasic approach showed that the strains belonged to the green-alga Acrochaete leptochaete, the diatom Nanofrustulum cf. shiloi, the rhodophyte Acrochaetium spongicola, and the chlorachniophyte Lotharella oceanica. A. leptochaete is reported for the first time in sponges, even though green algae are known to be associated with sponges. Nanofrustulum shiloi was found in association with the sponges Agelas oroides and Chondrilla nucula, whereas information existed only for its association with the species Aplysina aerophoba. Acrochaetium spongicola was found for the first time in association with sponges in the eastern Mediterranean. Moreover, we report herein for the first time a sponge-chlorarachniophycean association. Our research revealed new diversity of microalgae associated with sponges and added new records of sponge species, previously unknown for their association with microalgae.

Use of organic exudates from two polar diatoms by bacterial isolates from the Arctic Ocean.

Global warming affects primary producers in the Arctic, with potential consequences for the bacterial community composition through the consumption of microalgae-derived dissolved organic matter (DOM). To determine the degree of specificity in the use of an exudate by bacterial taxa, we used simple microalgae-bacteria model systems. We isolated 92 bacterial strains from the sea ice bottom and the water column in spring-summer in the Baffin Bay (Arctic Ocean). The isolates were grouped into 42 species belonging to Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. Forty strains were tested for their capacity to grow on the exudate from two Arctic diatoms. Most of the strains tested (78%) were able to grow on the exudate from the pelagic diatom Chaetoceros neogracilis, and 33% were able to use the exudate from the sea ice diatom Fragilariopsis cylindrus. 17.5% of the strains were not able to grow with any exudate, while 27.5% of the strains were able to use both types of exudates. All strains belonging to Flavobacteriia (n = 10) were able to use the DOM provided by C. neogracilis, and this exudate sustained a growth capacity of up to 100 times higher than diluted Marine Broth medium, of two Pseudomonas sp. strains and one Sulfitobacter strain. The variable bioavailability of exudates to bacterial strains highlights the potential role of microalgae in shaping the bacterial community composition. This article is part of the theme issue 'The changing Arctic Ocean: consequences for biological communities, biogeochemical processes and ecosystem functioning'.

Amyloid protein produced by B. cereus CR4 possesses bioflocculant activity and has potential application in microalgae harvest.

Bacillus cereus CR4 from the flocs of activated sludge was found to produce an extracellular bioflocculant, which was characterized as amyloid protein and demonstrated to have potential application in microalgae recovery. Cell surface amyloid production was demonstrated by fluorescence, confocal and scanning electron microscopy. Birefringence, spectral shift assay, TEM, FTIR and CD spectra confirmed the amyloid nature of the purified protein that demonstrated flocculation. The gene for amyloid protein of B. cereus CR4 was found to be related to tasA gene of amyloid protein produced by Bacillus subtilis. The results demonstrated that the amyloid protein produced by B. cereus CR4 possessed a novel bioflocculant activity which at pH below 4.5 reached to a maximum of 86.87%. The amyloid bioflocculant producing B. cereus CR4 has a potential in biotechnological application like Scenedesmus biomass recovery.

Improvement of carotenoid extraction from a recently isolated, robust microalga, Tetraselmis sp. CTP4 (chlorophyta).

In recent years, there has been increasing consumer interest in carotenoids, particularly of marine sustainable origin with applications in the food, cosmeceutical, nutritional supplement and pharmaceutical industries. For instance, microalgae belonging to the genus Tetraselmis are known for their biotechnologically relevant carotenoid profile. The recently isolated marine microalgal strain Tetraselmis sp. CTP4 is a fast-growing, robust industrial strain, which has successfully been produced in 100-m3 photobioreactors. However, there are no reports on total carotenoid contents from this strain belonging to T. striata/convolutae clade. Although there are several reports on extraction methods targeting chlorophytes, extraction depends on the strength of cell coverings, solvent polarity and the nature of the targeted carotenoids. Therefore, this article evaluates different extraction methods targeting Tetraselmis sp. CTP4, a strain known to contain a mechanically resistant theca. Here, we propose a factorial experimental design to compare extraction of total carotenoids from wet and freeze-dried microalgal biomass using four different solvents (acetone, ethanol, methanol or tetrahydrofuran) in combination with two types of mechanical cell disruption (glass beads or dispersion). The extraction efficiency of the methods was assessed by pigment contents and profiles present in the extracts. Extraction of wet biomass by means of glass bead-assisted cell disruption using tetrahydrofuran yielded the highest amounts of lutein and ß-carotene (622 ± 40 and 618 ± 32 µg g-1 DW, respectively). Although acetone was slightly less efficient than tetrahydrofuran, it is preferable due to its lower costs and toxicity.

Revision of Coelastrella (Scenedesmaceae, Chlorophyta) and first register of this green coccoid microalga for continental Norway.

A terrestrial green microalga was isolated at Ås, in Akershus County, Norway. The strain corresponded to a coccoid chlorophyte. Morphological characteristics by light and electron microscopy, in conjunction with DNA amplification and sequencing of the 18 s rDNA gene and ITS sequences, were used to identify the microalgae. The characteristics agree with those of the genus Coelastrella defined by Chodat, and formed a sister group with the recently described C. thermophila var. globulina. Coelastrella is a relatively small numbered genus that has not been observed in continental Norway before; there are no previous cultures available in collections of Norwegian strains. Gas chromatography analyses of the FAME-derivatives showed a high percentage of polyunsaturated fatty acids (44-45%) especially linolenic acid (C18:3n3; 30-34%). After the stationary phase, the cultures were able to accumulate several carotenoids as neoxanthin, pheophytin a, astaxanthin, canthaxanthin, lutein, and violaxanthin. Due to the scarcity of visual characters suitable for diagnostic purposes and the lack of DNA sequence information, there is a high possibility that species of this genus have been neglected in local environmental studies, even though it showed interesting properties for algal biotechnology.

Dielectrophoretic separation of microalgae cells in ballast water in a microfluidic chip.

The composition of the ship's ballast water is complex and contains a large number of microalgae cells, bacteria, microplastics, and other microparticles. To increase the accuracy and efficiency of detection of the microalgae cells in ballast water, a new microfluidic chip for continuous separation of microalgae cells based on alternating current dielectrophoresis was proposed. In this microfluidic chip, one piece of 3-dimensional electrode is embedded on one side and eight discrete electrodes are arranged on the other side of the microchannel. An insulated triangular structure between electrodes is designed for increasing the inhomogeneity of the electric field distribution and enhancing the dielectrophoresis (DEP) force. A sheath flow is designed to focus the microparticles near the electrode, so as to increase the suffered DEP force and improve separation efficiency. To demonstrate the performance of the microfluidic separation chip, we developed two species of microalgae cells (Platymonas and Closterium) and a kind of microplastics to be used as test samples. Analyses of the related parameters and separation experiments by our designed microfluidic chip were then conducted. The results show that the presented method can separate the microalgae cells from the mixture efficiently, and this is the first time to separate two or more species of microalgae cells in a microfluidic chip by using negative and positive DEP force simultaneously, and moreover it has some advantages including simple operation, high efficiency, low cost, and small size and has great potential in on-site pretreatment of ballast water.

Rapid and sensitive detection of Amphidinium carterae by loop-mediated isothermal amplification combined with a chromatographic lateral-flow dipstick.

Frequent outbreaks of toxic algal blooms devastate marine ecosystems, marine fisheries, and public health. Monitoring toxic algae is crucial to reduce losses caused by imminent algal blooms. However, traditional detection techniques relying on morphological examination are tedious and time-consuming. Therefore, the development of convenient strategies to detect toxin-producing microalgae is necessary. In this study, a novel method for the rapid, sensitive detection of Amphidinium carterae by loop-mediated isothermal amplification (LAMP) combined with a chromatographic lateral-flow dipstick (LFD) was established. The partial internal transcribed spacer gene was PCR amplified, cloned, and sequenced to design four LAMP primers and a detection probe for A. carterae detection. The LAMP detection conditions were optimized, and the optimum parameters were determined to be the following: dNTP concentration, 1.2 mM; betaine concentration, 1.2 M; magnesium ion concentration, 8 mM; ratio of inner primer to outer primer, 8:1; amplification temperature, 59 °C; and amplification time, 60 min. The specificity of LAMP-LFD was confirmed by cross-reactivity tests with other algal species. LAMP-LFD was 100 times more sensitive than regular PCR and similarly sensitive as LAMP and SYBR Green I. LAMP-LFD can be completed within 70 min and did not require special detection equipment. The convenience of the established LAMP-LFD assay was further validated by tests with simulated field-water samples. In conclusion, the developed LAMP-LFD assay can be used as a reliable and simple method of detecting A. carterae.

The selection of a surfactant for freshwater microalgae harvesting and separation by the foam separation method.

Collecting microalgae from water with less energy and cost is significant to gain economic profit from microalgae harvesting and processing. Foam separation has certain advantages including low energy consumption, simple operation and easy maintenance of the equipment. Natural surfactants, compared to traditional surfactants, were used to harvest and separate the freshwater microalgae Desmodesmus brasiliensis by foam separation. Results showed a recovery percentage of 93.6% and an enrichment ratio of 23.1 with the natural surfactant cocamidopropyl betaine (CAPB), suggesting that this low-cost surfactant can be applied to microalgae biomass recovery on a commercial scale using foam separation with no negative effect on the content of microalgae chlorophyll, carotenoid or protein.

Evaluation of Chlorella sorokiniana isolated from local municipal wastewater for dual application in nutrient removal and biodiesel production.

The isolated microalga Chlorella sorokiniana BENHA721_ABO4 was grown in Bold's basal medium (BBM) as a control, municipal wastewater (WW), and wastewater enriched with BBM elements (WW+). Cultivation in WW+ showed the highest cell number which represented 25.3 and 47.3% over that grown in WW and BBM, respectively. However, rapid growth in WW+ was accompanied by significant reduction in lipid content. Due to lipid accumulation in WW, it showed the maximum significant lipid productivity of 16.2 mg L-1 day-1. Microalgae cultivation in WW for 10 days showed 74.2, 83.3, and 78.0% removal efficiency for NO3-N, NH3-N and TP, respectively. In addition, growth in WW significantly reduced polyunsaturated fatty acids by 36.0% with respect to BBM in favor of monounsaturated fatty acids. The present results confirmed that C. sorokiniana isolate BENHA721_ABO4 grown in secondary effluent municipal wastewater offers real potential for future application in wastewater treatment and biodiesel production.

Growth and metabolic characteristics of oleaginous microalgal isolates from Nilgiri biosphere Reserve of India.

BACKGROUND: Renewable energy for sustainable development is a subject of a worldwide debate since continuous utilization of non-renewable energy sources has a drastic impact on the environment and economy; a search for alternative energy resources is indispensable. Microalgae are promising and potential alternate energy resources for biodiesel production. Thus, our efforts were focused on surveying the natural diversity of microalgae for the production of biodiesel. The present study aimed at identification, isolation, and characterization of oleaginous microalgae from shola forests of Nilgiri Biosphere Reserve (NBR), the biodiversity hot spot of India, where the microalgal diversity has not yet been systematically investigated. RESULTS: Overall the higher biomass yield, higher lipid accumulation and thermotolerance observed in the isolated microalgal strains have been found to be the desirable traits for the efficient biodiesel production. Species composition and diversity analysis yielded ten potential microalgal isolates belonging to Chlorophyceae and Cyanophyceae classes. The chlorophytes exhibited higher growth rate, maximum biomass yield, and higher lipid accumulation than Cyanophyceae. Among the chlorophytes, the best performing strains were identified and represented by Acutodesmus dissociatus (TGA1), Chlorella sp. (TGA2), Chlamydomonadales sp. (TGA3) and Hindakia tetrachotoma (PGA1). The Chlamydomonadales sp. recorded with the highest growth rate, lipid accumulation and biomass yield of 0.28 ± 0.03 day-1 (µexp), 29.7 ± 0.69% and 134.17 ± 16.87 mg L-1 day-1, respectively. It was also found to grow well at various temperatures, viz., 25 °C, 35 °C, and 45 °C, indicating its suitability for open pond cultivation. The fatty acid methyl ester (FAME) analysis of stationary phase cultures of selected four algal strains by tandem mass spectrograph showed C16:0, C18:1 and C18:3 as dominant fatty acids suitable for biodiesel production. All the three strains except for Hindakia tetrachotoma (PGA1) recorded higher carbohydrate content and were considered as potential feed stocks for biodiesel production through hydrothermal liquefaction technology (HTL). CONCLUSIONS: In conclusion, the present investigation is a first systematic study on the microalgal diversity of soil and water samples from selected sites of NBR. The study resulted in isolation and characterization of ten potent oleaginous microalgae and found four cultures as promising feed stocks for biodiesel production. Of the four microalgae, Chlamydomonadales sp. (TGA3) was found to be significantly thermo-tolerant and can be considered as promising feedstock for biodiesel production.

Aglaophenia octodonta (Cnidaria, Hydrozoa) and the Associated Microbial Community: a Cooperative Alliance?

Recently, genetic approaches have revealed a surprising bacterial world as well as a growing knowledge of the enormous distribution of animal-bacterial interactions. In the present study, the diversity of the microorganisms associated to the hydroid Aglaophenia octodonta was studied with epifluorescence, optical, and scanning electron microscopy. Small subunit ribosomal RNA gene sequencing with "universal" and taxon-specific primers allowed the assignment of the microalgae to Symbiodinium and the peritrich ciliates to Pseudovorticella, while the luminous vibrios were identified as Vibrio jasicida of the Harvey clade. To understand the possible relationships among Vibrio jasicida, Symbiodinium, A. octodonta, and Pseudovorticella, specific treatments were conducted in microcosm experiments, with the antibiotic ampicillin and other substances that interfere with bacterial and hydroid metabolism. Treatment of A. octodonta with ampicillin resulted in a decrease of bacterial luminescence followed by Pseudovorticella detachment and Symbiodinium expulsion and suggesting that these microorganisms form a "consortium" with beneficial metabolic interdependence. This hypothesis was reinforced by the evidence that low concentrations of hydrogen peroxide, which stimulate the bacterial oxidative metabolism and luminescence by releasing oxygen, were able to counteract the detrimental effect of ampicillin on the stability of the studied A. octodonta association. A model is proposed in which microalgae that release oxygen during photosynthesis are useful to luminous bacteria for their metabolism and for establishing/maintaining symbiosis leading to a close alliance and mutual benefit of the system A. octodonta-Vibrio jasicida-Pseudovorticella sp.-Symbiodinium sp.

Exploiting diversity and synthetic biology for the production of algal biofuels.

Modern life is intimately linked to the availability of fossil fuels, which continue to meet the world's growing energy needs even though their use drives climate change, exhausts finite reserves and contributes to global political strife. Biofuels made from renewable resources could be a more sustainable alternative, particularly if sourced from organisms, such as algae, that can be farmed without using valuable arable land. Strain development and process engineering are needed to make algal biofuels practical and economically viable.

Cultivation of newly isolated microalgae Coelastrum sp. in wastewater for simultaneous CO2 fixation, lipid production and wastewater treatment.

Cultivation of microalgae in wastewater is a promising and cost-effective approach for both CO2 biofixation and wastewater remediation. In this study, a new strain of Coelastrum sp. was isolated from cattle manure leachate. The isolated microalgae were then cultivated in wastewater. Effects of different sCOD concentrations (600, 750, 900, 1050 mg L-1) and light intensities (1000, 2300, 4600, 6900 and 10000 Lux) on biomass production, CO2 consumption rate and nutrient removal from wastewater were investigated. The results showed that maximum cell growth and CO2 consumption rate were 2.71 g L-1 and 53.12 mg L-1 day-1, respectively, which were obtained in the wastewater with 750 mg L-1 sCOD and under the light intensity of 6900 Lux. The microalgae were able to completely consume all CO2 after incubation period of 4 days. The highest sCOD, total Kjeldahl nitrogen (TKN), nitrate and total phosphorous (TP) removal at such conditions were 53.45, 91.18, 87.51 and 100%, respectively. The lipid content of microalgal biomass was also measured under different light intensities; maximum amount of lipid was determined to be 50.77% under illumination of 2300 Lux. Finally, the CO2 consumption rate and biomass productivity of microalgae in semi-batch culture with continuous gas flow (CO2 6%:N2 94%) were investigated. The rate of CO2 consumption and biomass productivity were 0.528 and 0.281 g L-1 day-1, respectively. The TKN, nitrate, TP and sCOD removal rate of microalgae were 83.51, 80.91, 100, 41.4%, respectively.