Search results for: heterotrophic denitrification

Authors: Etleva Hamzaraj, Eva Kica, Anila Paparisto, Pranvera Lazo

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Worldwide demographic developments of recent decades have been associated with negative environmental consequences. For this reason, there is a growing interest in assessing the state of natural ecosystems or assessing human impact on them. In this respect, this study aims to evaluate the change in water quality of the Mat River for a period of about ten years to highlight human impact. In one year, period of study, several biological and environmental parameters are determined to evaluate river water quality, and the data collected are compared with those of a similar study in 2007. Samples are collected every month in five stations evenly distributed along the river. Total coliform bacteria, the number of heterotrophic bacteria in water, and benthic macroinvertebrates are used as biological parameters of water quality. The most probable number index is used for evaluation of total coliform bacteria in water, while the number of heterotrophic bacteria is determined by counting colonies on plates with Plate Count Agar, cultivated with 0.1 ml sample after series dilutions. Benthic macroinvertebrates are analyzed by the number of individuals per taxa, the value of biotic index, EPT Richness Index value and tolerance value. Environmental parameters like pH, temperature, and electrical conductivity are measured onsite. As expected, the bacterial load was higher near urban areas, and the pollution increased with the course of the river. The maximum concentration of fecal coliforms was 1100 MPN/100 ml in summer and near the most urbanized area of the river. The data collected during this study show that after about ten years, there is a change in water quality of Mat River. According to a similar study carried out in 2007, the water of Mat River was of ‘excellent’ quality. But, according to this study, the water was classified as of ‘excellent’ quality only in one sampling site, near river source, while in all other stations was of ‘good’ quality. This result is based on biological and environmental parameters measured. The human impact on the quality of water of Mat River is more than evident.

Keywords: water quality, coliform bacteria, MPN index, benthic macroinvertebrates, biotic index

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Authors: P. Kiran Kumar, S. Vijaya Krishna, Kavita Verma1, V. Himabindu

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Concern about global warming and energy security has led to increased biomass utilization as an alternative feedstock to fossil fuels. Biomass is a promising feedstock since it is abundant and cheap and can be transformed into fuels and chemical products. Microalgae biofuels are likely to have a much lower impact on the environment. Microalgae cultivation using sewage with industrial flue gases is a promising concept for integrated biodiesel production, CO₂ sequestration, and nutrients recovery. Autotrophic, Mixotrophic, and Heterotrophic are the three modes of cultivation for microalgae biomass. Several mechanical and chemical processes are available for the extraction of lipids/oily components from microalgae biomass. In organic solvent extraction methods, a prior drying of biomass and recovery of the solvent is required, which are energy-intensive. Thus, the hydrothermal process overcomes the drawbacks of conventional solvent extraction methods. In the hydrothermal process, the biomass is converted into oily components by processing in a hot, pressurized water environment. In this process, in addition to the lipid fraction of microalgae, other value-added products such as proteins, carbohydrates, and nutrients can also be recovered. In the present study was (Scenedesmus quadricauda) was isolated and cultivated in autotrophic, heterotrophic, and mixotrophically using sewage wastewater and industrial flue gas in batch and continuous mode. The harvested algae biomass from S. quadricauda was used for the recovery of lipids and bio-oil. The lipids were extracted from the algal biomass using sonication as a cell disruption method followed by solvent (Hexane) extraction, and the lipid yield obtained was 8.3 wt% with Palmitic acid, Oleic acid, and Octadeonoic acid as fatty acids. The hydrothermal process was also carried out for extraction of bio-oil, and the yield obtained was 18wt%. The bio-oil compounds such as nitrogenous compounds, organic acids, and esters, phenolics, hydrocarbons, and alkanes were obtained by the hydrothermal process of algal biomass. Nutrients such as NO₃⁻ (68%) and PO₄⁻ (15%) were also recovered along with bio-oil in the hydrothermal process.

Keywords: flue gas, hydrothermal process, microalgae, sewage wastewater, sonication

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Authors: M. T. Amina, A. A. Alazba, U. Manzoor

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Solar disinfection (SODIS) is a viable method for household water treatment and is recommended by the World Health Organization as cost effective approach that can be used without special skills. The efficiency of both SODIS and solar collector disinfection (SOCODIS) system was evaluated using four different sources of water including stored rainwater, storm water, ground water and treated sewage. Samples with naturally occurring microorganisms were exposed to sunlight for about 8-9 hours in 2-L polyethylene terephthalate bottles under similar experimental conditions. Total coliform (TC), Escherichia coli (E. coli) and heterotrophic plate counts (HPC) were used as microbial water quality indicators for evaluating the disinfection efficiency at different sunlight intensities categorized as weak, mild and strong weathers. Heterotrophic bacteria showed lower inactivation rates compared to E. coli and TC in both SODIS and SOCODIS system. The SOCODIS system at strong weather was the strongest disinfection system in this study and the complete inactivation of HPC was observed after 8-9 hours of exposure with SODIS being ineffective for HPC. At moderate weathers, however, the SOCODIS system did not show complete inactivation of HPC due to very high concentrations (up to 5x10^7 CFU/ml) in both storm water and treated sewage. SODIS even remained ineffective for the complete inactivation of E. coli due to its high concentrations of about 2.5x10^5 in treated sewage compared with other waters even after 8-9 hours of exposure. At weak weather, SODIS was not effective at all while SOCODIS system, though incomplete, showed good disinfection efficiency except for HPC and to some extent for high E. coli concentrations in storm water. Largest reduction of >5 log occurred for TC when used stored rainwater even after 6 hours of exposure in the case of SOCODIS system at strong weather. The lowest E. coli and HPC reduction of ~2 log was observed in SODIS system at weak weather. Further tests with varying pH and turbidity are required to understand the effects of reaction parameters that could be a step forward towards maximizing the disinfection efficiency of such systems for the complete inactivation of naturally occurring E. coli or HPC at moderate or even at weak weathers.

Keywords: efficiency, microbial, SODIS, SOCODIS, weathers

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Authors: Anastasia Poliakova

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Harmful algal bloom (HAB) is a known negative phenomenon that is caused both by natural factors and anthropogenic influence. HABs can result in a series of deleterious effects, such as beach fouling, paralytic shellfish poisoning, mass mortality of marine species, and a threat to human health, especially if toxins pollute drinking water or occur nearby public resorts. In South China, the problem of HABs has an ultimately important meaning. For this study, we used a 1.5 m sediment core LAX-2018-2 collected in 2018 from the Zhanjiang Mangrove National Nature Reserve (109°03´E, 20°30´N), Guangdong Province, South China. High-resolution coastal environment reconstruction with a specific focus on the HABs history during the last ca 2500 yrs was attempted. Age control was performed with five radiocarbon dates obtained from benthic foraminifera. A total number of 71 dinoflagellate cyst types was recorded. The most common types found consistently throughout the sediment sequence were autotrophic Spiniferites spp., Spiniferites hyperacanthus and S. mirabilis, S. ramosus, Operculodinium centrocarpum sensu Wall and Dale 1966, Polysphaeridium zoharyi, and heterotrophic Brigantedinium ssp., cyst of Gymnodinium catenatum and cysts mixture of Protoperidinium. Three local dinoflagellate zones LAX-1 to LAX-3 were established based on the results of the constrained cluster analysis and data ordination; additionally, the middle zone LAX-2 was derived into two subzones, LAX-2a and LAX-2b based on the dynamics of toxic and heterotrophic cysts as well as on the significant changes (probability, P=0.89) in percentages of eutrophic indicators. The total cyst count varied from 106 to 410 dinocysts per slide, with 177 cyst types on average. Dinocyst assemblages are characterized by high values of the dost-depositional degradation index (kt) that varies between 3.6 and 7.6 (averaging 5.4), which is relatively high and is very typical for the areas with selective dinoflagellate cyst preservation that is related to bottom-water oxygen concentrations.

Keywords: reconstruction of palaeoenvironment, harmful algal blooms, anthropogenic influence on coastal zones, South China Sea

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Authors: Hsin-Yueh Chang, Pin-Chen Liao, Jo-Shu Chang, Chun-Yen Chen

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Biofuel has drawn much attention as a potential substitute to fossil fuels. However, biodiesel from waste oil, oil crops or other oil sources can only satisfy partial existing demands for transportation. Due to the feature of being clean, green and viable for mass production, using microalgae as a feedstock for biodiesel is regarded as a possible solution for a low-carbon and sustainable society. In particular, Thraustochytrium sp. BM2, an indigenous heterotrophic microalga, possesses the potential for metabolizing glycerol to produce lipids. Hence, it is being considered as a promising microalgae-based oil source for biodiesel production and other applications. This study was to optimize the culture pH, scale up, assess the feasibility of producing microalgal lipid from crude glycerol and apply operation strategies following optimal results from shake flask system in a 5L stirred-tank fermenter for further enhancing lipid productivities. Cultivation of Thraustochytrium sp. BM2 without pH control resulted in the highest lipid production of 3944 mg/L and biomass production of 4.85 g/L. Next, when initial glycerol and corn steep liquor (CSL) concentration increased five times (50 g and 62.5 g, respectively), the overall lipid productivity could reach 124 mg/L/h. However, when using crude glycerol as a sole carbon source, direct addition of crude glycerol could inhibit culture growth. Therefore, acid and metal salt pretreatment methods were utilized to purify the crude glycerol. Crude glycerol pretreated with acid and CaCl₂ had the greatest overall lipid productivity 131 mg/L/h when used as a carbon source and proved to be a better substitute for pure glycerol as carbon source in Thraustochytrium sp. BM2 cultivation medium. Engineering operation strategies such as fed-batch and semi-batch operation were applied in the cultivation of Thraustochytrium sp. BM2 for the improvement of lipid production. In cultivation of fed-batch operation strategy, harvested biomass 132.60 g and lipid 69.15 g were obtained. Also, lipid yield 0.20 g/g glycerol was same as in batch cultivation, although with poor overall lipid productivity 107 mg/L/h. In cultivation of semi-batch operation strategy, overall lipid productivity could reach 158 mg/L/h due to the shorter cultivation time. Harvested biomass and lipid achieved 232.62 g and 126.61 g respectively. Lipid yield was improved from 0.20 to 0.24 g/g glycerol. Besides, product costs of three kinds of operation strategies were also calculated. The lowest product cost 12.42 $NTD/g lipid was obtained while employing semi-batch operation strategy and reduced 33% in comparison with batch operation strategy.

Keywords: heterotrophic microalga Thrasutochytrium sp. BM2, microalgal lipid, crude glycerol, fermentation strategy, biodiesel

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Authors: Michael Huber, Maximilian J. Poller, Jens Tochtermann, Wolfgang Korth, Andreas Jess, Jakob Albert

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Aside from the desulfurisation, the denitrogenation of fuels is of great importance to minimize the environmental impact of transport emissions. The oxidative reaction pathway of organic nitrogen in the catalytic oxidative denitrogenation could be successfully elucidated. This is the first time such a pathway could be traced in detail in non-microbial systems. It was found that the organic nitrogen is first oxidized to nitrate, which is subsequently reduced to molecular nitrogen via nitrous oxide. Hereby, the organic substrate serves as a reducing agent. The discovery of this pathway is an important milestone for the further development of fuel denitrogenation technologies. The United Nations aims to counteract global warming with Net Zero Emissions (NZE) commitments; however, it is not yet foreseeable when crude oil-based fuels will become obsolete. In 2021, more than 50 million barrels per day (mb/d) were consumed for the transport sector alone. Above all, heteroatoms such as sulfur or nitrogen produce SO₂ and NOx during combustion in the engines, which is not only harmful to the climate but also to health. Therefore, in refineries, these heteroatoms are removed by hy-drotreating to produce clean fuels. However, this catalytic reaction is inhibited by the basic, nitrogenous reactants (e.g., quinoline) as well as by NH3. The ion pair of the nitrogen atom forms strong pi-bonds to the active sites of the hydrotreating catalyst, which dimin-ishes its activity. To maximize the desulfurization and denitrogenation effectiveness in comparison to just extraction and adsorption, selective oxidation is typically combined with either extraction or selective adsorption. The selective oxidation produces more polar compounds that can be removed from the non-polar oil in a separate step. The extraction step can also be carried out in parallel to the oxidation reaction, as a result of in situ separation of the oxidation products (ECODS; extractive catalytic oxidative desulfurization). In this process, H8PV5Mo7O40 (HPA-5) is employed as a homogeneous polyoxometalate (POM) catalyst in an aqueous phase, whereas the sulfur containing fuel components are oxidized after diffusion from the organic fuel phase into the aqueous catalyst phase, to form highly polar products such as H₂SO₄ and carboxylic acids, which are thereby extracted from the organic fuel phase and accumulate in the aqueous phase. In contrast to the inhibiting properties of the basic nitrogen compounds in hydrotreating, the oxidative desulfurization improves with simultaneous denitrification in this system (ECODN; extractive catalytic oxidative denitrogenation). The reaction pathway of ECODS has already been well studied. In contrast, the oxidation of nitrogen compounds in ECODN is not yet well understood and requires more detailed investigations.

Keywords: oxidative reaction pathway, denitrogenation of fuels, molecular catalysis, polyoxometalate

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Authors: Sakura Yoshii, Masakazu Abe, Akihiro Iijima

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Nitrogen saturation has become one of the serious issues in the field of forest environment. The watershed protection forests located in the downwind hinterland of Tokyo Metropolitan Area are believed to be facing nitrogen saturation. In this study, we carefully focus on the balance of nitrogen between load and runoff. Annual nitrogen load via atmospheric deposition was estimated to 461.1 t-N/year in the upper reaches of the Kanna River. Annual nitrogen runoff to the forested headwater stream of the Kanna River was determined to 184.9 t-N/year, corresponding to 40.1% of the total nitrogen load. Clear seasonal change in NO3-N concentration was still observed. Therefore, watershed protection forest of the Kanna River is most likely to be in Stage-1 on the status of nitrogen saturation.

Keywords: atmospheric deposition, nitrogen accumulation, denitrification, forest ecosystems

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Authors: Gabriela Kalcikova, Igor Bosevski, Ula Rozman, Andreja Zgajnar Gotvajn

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Antibiotics are extensively used in human medicine and also in animal husbandry to prevent or control infections. Recently, a lot of attention has been put on veterinary antibiotics, because their global consumption is increasing and it is expected to be 106.600 tons in 2030. Most of veterinary antibiotics are introduced into the environment via animal manure, which is used as fertilizer. One of such veterinary antibiotics is tiamulin. It is used the form of fumarate for treatment of pig and poultry. It is used against prophylaxis of dysentery, pneumonia and mycroplasmal infections, but its environmental impact is practically unknown. Tiamulin has been found very persistent in animal manure and thus it is expected that can be, during rainfalls, transported into the aquatic environment and affect various organisms. For assessment of its environmental impact, it is necessary to evaluate its biodegradability and toxicity to various organisms from different levels of a food chain. Therefore, the aim of our study was to evaluate ready biodegradability and toxicity of tiamulin fumarate to various organisms. Bioassay used included luminescent bacterium Vibrio fischeri heterotrophic and nitrifying microorganisms of activated sludge, water flea Daphnia magna and duckweed Lemna minor. For each species, EC₅₀ values were calculated. Biodegradability test was used for determination of ready biodegradability and it provides information about biodegradability of tiamulin under the most common environmental conditions. Results of our study showed that tiamulin differently affects selected organisms. The most sensitive organisms were water fleas with 48hEC₅₀ = 14.2 ± 4.8 mg/L and duckweed with 168hEC₅₀ = 22.6 ± 0.8 mg/L. Higher concentrations of tiamulin (from 10 mg/L) significantly affected photosynthetic pigments content in duckweed and concentrations above 80 mg/L cause visible chlorosis. It is in agreement with previous studies showing significant effect of tiamulin on green algae and cyanobacteria. Tiamuline has a low effect on microorganisms. The lower toxicity was observed for heterotrophic microorganisms (30minEC₅₀ = 1656 ± 296 mg/L), than Vibrio fisheri (30minEC₅₀ = 492 ± 21) and the most sensitive organisms were nitrifying microorganisms (30minEC₅₀ = 183 ± 127 mg/L). The reason is most probably the mode of action of tiamulin being effective to gram-positive bacteria while gram-negative (e.g., Vibrio fisheri) are more tolerant to tiamulin. Biodegradation of tiamulin was very slow with a long lag-phase being 20 days. The maximal degradation reached 40 ± 2 % in 43 days of the test and tiamulin as other antibiotics (e.g. ciprofloxacin) are not easily biodegradable. Tiamulin is widely used antibiotic in veterinary medicine and thus present in the environment. According to our results, tiamulin can have negative effect on water fleas and duckweeds, but the concentrations are several magnitudes higher than that found in any environmental compartment. Tiamulin is low toxic to tested microorganisms, but it is very low biodegradable and thus possibly persistent in the environment.

Keywords: antibiotics, biodegradability, tiamulin, toxicity

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Authors: Junior B. N. Adohinzin, Ling Xu

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Nitrogen is among the main nutrients encouraging the growth of organic matter and algae which cause eutrophication in water bodies. Therefore, its removal from wastewater has become a worldwide emerging concern. In this research, an innovative Membrane Bioreactor (MBR) system named “moving bed membrane bioreactor (MBMBR)” was developed and investigated under intermittently-aerated mode for simultaneous removal of organic carbon and nitrogen. Results indicated that the variation of the intermittently aerated duration did not have an apparent impact on COD and NH4+–N removal rate, yielding the effluent with average COD and NH4+–N removal efficiency of more than 92 and 91% respectively. However, in the intermittently aerated cycle of (continuously aeration/0s mix), (aeration 90s/mix 90s) and (aeration 90s/mix 180s); the average TN removal efficiency was 67.6%, 69.5% and 87.8% respectively. At the same time, their nitrite accumulation rate was 4.5%, 49.1% and 79.4% respectively. These results indicate that the intermittently aerated mode is an efficient way to controlling the nitrification to stop at nitrition; and also the length of anoxic duration is a key factor in improving TN removal.

Keywords: membrane bioreactor (MBR), moving bed biofilm reactor (MBBR), nutrients removal, simultaneous nitrification and denitrification

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Authors: S. K. Al-Musharafi, I. Y. Mahmoud, S. N. Al-Bahry

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At Ras Al-Hadd Reserve, Eggs from green turtles and Chelonia mydas were randomly collected immediately after Oviposition. Eggshells taken from fresh eggs and sand collected from the body chamber were analyzed for eight heavy metals (Al, Br, Cd, Co, Cu, Fe, S, and Zn) using inductively coupled plasma mass spectrometry (ICP). Heavy metal concentrations varied significantly (P<0.05) between nest sand and eggshells. Zn values were significantly higher than the other heavy metals. A total of 60 heterotrophic bacteria belong to eight genera were isolated from fresh egg contents (albumen and yolk). Resistance of the isolates to Amikacin, ampicillin, chloramphenicol, gentamycine, minocylin, nalidixicacid, neomycin, streptomycin, tetracycline, tobramycin, and Trimethoprim was tested. More than 40 % of the isolates were multiple resistant to 2-7 antibiotics. Most of the resistant strains were also resistant to Zn. The value of these findings may indicate that the origin of pollution is of human contaminated effluents.

Keywords: antibiotic resistance, bacteria, environment, heavy metals, sea turtles

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Authors: Fezzani Boubaker

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In this work, a comprehensive study of respirometric method was performed to assess the biodegradable COD fractions of domestic sewage mixed with baker’s yeast factory effluent treated by wastewater treatment plant (WWTP) of Beja. Three respirometric runs were performed in a closed tank reactor to characterize this mixed raw effluent. Respirometric result indicated that the readily biodegradable fraction (SS) was in range of 6-22%, the slowly biodegradable fraction (Xs) was in range of 33-42%, heterotrophic biomass (XH) was in range of 9-40% and the inert fractions: XI and SI were in range of 2-40% and 6-12% respectively which were high due to the presence of baker’s yeast factory effluent compared to domestic effluent alone. The fractions of the total nitrogen showed that SNO fraction is between 6 and 9% of TKN, the fraction of nitrogen ammonia SNH was ranging from 5 to 68%. The organic fraction divided into two compartments SND (11-85%) and XND (5-20%) the inert particulate nitrogen fraction XNI was between 0.4 and 1% and the inert soluble fraction of nitrogen SNI was ranged from 0.4 to 3%.

Keywords: wastewater characterization, COD fractions, respirometry, domestic sewage

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Authors: C. E. Ihejirika, M. I. Nwachukwu, R. F. Njoku-Tony, O. C. Ihejirika, U. O. Enwereuzoh, E. O. Imo, D. C. Ashiegbu

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Disposal of industrial solid wastes in the environment is a major environmental challenge. This study investigated the effects of calcium carbide waste dumpsites on soil quality. Soil samples were collected with hand auger from three different dumpsites at varying depths and made into composite samples. Samples were subjected to standard analytical procedures. pH varied from 10.38 to 8.28, nitrate from 5.6mg/kg to 9.3mg/kg, phosphate from 8.8mg/kg to 12.3mg/kg, calcium carbide reduced from 10% to to 3%. Calcium carbide was absent in control soil samples. Bacterial counts from dumpsites ranged from 1.8 x 105cfu/g - 2.5 x 105cfu/g while fungal ranged from 0.8 x 103cfu/g - 1.4 x 103cfu/g. Bacterial isolates included Pseudomonas spp, Flavobacterium spp, and Achromobacter spp, while fungal isolates include Penicillium notatum, Aspergillus niger, and Rhizopus stolonifer. No organism was isolated from the dumpsites at soil depth of 0-15 cm, while there were isolates from other soil depths. Toxicity might be due to alkaline condition of the dumpsite. Calcium carbide might be bactericidal and fungicidal leading to cellular physiology, growth retardation, death, general loss of biodiversity and reduction of ecosystem processes. Detoxification of calcium carbide waste before disposal on soil might be the best option in management.

Keywords: biodiversity, calcium-carbide, denitrification, toxicity

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Authors: B. A. A. Paixão, J. V. B. Gomide

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This work aims to analyze the locative structure used by the locative games of the company Niantic. To fulfill this objective, a literature review on the representation and simulation of cities was developed; interviews with Ingress players and playing Ingress. Relating these data, it was possible to deepen the relationship between the virtual and the real to create the simulation of cities and their cultural objects in locative games. Cities representation associates geo-location provided by the Global Positioning System (GPS), with augmented reality and digital image, and provides a new paradigm in the city interaction with its parts and real and virtual world elements, homeomorphic to real world. Bibliographic review of papers related to the representation and simulation study and their application in locative games was carried out and is presented in the present paper. The cities representation and simulation concepts in locative games, and how this setting enables the flow and immersion in urban space, are analyzed. Some examples of games are discussed for this new setting development, which is a mix of real and virtual world. Finally, it was proposed a Locative Structure for electronic games using the concepts of heterotrophic representations and isotropic representations conjoined with immediacy and hypermediacy.

Keywords: cities representation, cities simulation, games simulation, immersion, locative games

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Authors: Hamed A. El-Serehy, Khaled A. Al-Rasheid, Fahad A Al-Misned

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The meiofaunal population fluctuation, microbial dynamic and the composition of the sedimentary organic matter were investigated seasonally in the Egyptian shores along the northern part of Red Sea. Total meiofaunal population densities were extremely low with an annual average of 109 ±26 ind./10 cm2 and largely dominated by nematodes (on annual average from 52% to 94% of total meiofaunal density). The benthic microbial population densities ranged from 0.26±0.02 x 108 to 102.67±18.62 x 108/g dry sediment. Total sedimentary organic matter concentrations varied between 5.8 and 11.6 mg/g and the organic carbon, which was measured as summation of the carbohydrates, proteins and lipids, accounted for only a small fraction of being 32 % of the total organic matter. Chlorophyll a attained very low values and fluctuated between 2 and 11 µg/g. The very low chlorophyll a concentration in the Egyptian coasts along the Red Sea can suggest that the sedimentary organic matter along the Egyptian coasts is dominated by organic detrital and heterotrophic bacteria on one hand, and do not promote carbon transfer towards the higher trophic level on the other hand. However, the present study indicates that the existing of well diversified meiofaunal group, with a total of ten meiofaunal taxa, can serve as food for higher trophic levels in the Red Sea marine ecosystem.

Keywords: bacteria, meiofauna, intertidal sediments, Red Sea

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Authors: E. Osazee, L. U. Bashir

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This study was conducted at the Department of Biological Sciences, Usmanu Danfodiyo University, Sokoto, Nigeria, to determine the effects of different weights of cow dung on indigenous bacterial isolates in remediation of crude oil contaminated soils. The soil (1kg) was contaminated with 20g of crude oil and this was treated with three (40g, 80g and 120g) weights of cow dung. The soils were amended after two weeks of crude oil contamination. Soil samples were collected from the plastic bags for microbiological analyses. The isolates were cultured to test their ability to grow on crude oil. The ability of the isolates to utilize the crude oil was determined using media dilution technique. Bacteria such as Proteus mirabilis, Bacillus lacterosporus, Morganella morganii, Serratia marcescens and Bacillus alvei were isolated. The variables measured were heterotrophic bacterial populations, hydrocarbon utilizing bacterial populations and the percentage of crude oil degraded in the soils. Data collected were subjected to analysis of variance (ANOVA). Results obtained indicated that all the different weights of cow dung showed appreciable effect in crude oil decontamination. Based on the findings of the experiments, it could be deduced that 120g of cow dung promoted higher degradation of hydrocarbons. Thus, it should be recommended for remediation of crude oil contaminated soil in the study area.

Keywords: crude oil, cow dung, amendment, bioremediation, decontamination

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Authors: A. Mohammed, A. Babatunde

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This study investigated the integrated removal of heavy metals, organic matter and nitrogen from landfill leachate using a novel laboratory scale constructed wetland system. The main objectives of this study were: (i) to assess the overall effectiveness of the constructed wetland system for treating landfill leachate; (ii) to examine the interactions and impact of key leachate constituents (heavy metals, organic matter and nitrogen) on the overall removal dynamics and efficiency. The constructed wetland system consisted of four stages operated in tidal flow and anoxic conditions. Results obtained from 215 days of operation have demonstrated extraordinary heavy metals removal up to 100%. Analysis of the physico- chemical data reveal that the controlling factors for metals removal were the anoxic condition and the use of the novel media (dewatered ferric sludge which is a by-product of drinking water treatment process) as the main substrate in the constructed wetland system. Results show that the use of the ferric sludge enhanced heavy metals removal and brought more flexibility to simultaneous nitrification and denitrification which occurs within the microbial flocs. Furthermore, COD and NH₄-N were effectively removed in the system and this coincided with enhanced aeration in the 2nd and 3rd stages of the constructed wetland system. Overall, the results demonstrated that the ferric dewatered sludge constructed wetland system would be an effective solution for integrated removal of pollutants from landfill leachates.

Keywords: constructed wetland, ferric dewatered sludge, heavy metals, landfill leachate

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Authors: Jian Li

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Carotenoids area group of metabolites in mostly photosynthetic organisms such as plants and microalgae and have wide applications in cosmetics, food, feed, and health industries. Although phototrophic cultivation of microalgae has been developed to produce some carotenoids for decades, most carotenoids are currently synthesized chemically at industrial scales because of affordable production costs. Chemical carotenoids are regarded not as safe for human beings as natural carotenoids and are restricted only for animal feed markets, and the industries call for inexpensive sources of natural products. Microalgae grow much quicker in mixotrophy than in phototrophy, and thus mixotrophic cultivation processes have great potential to reduce the production cost of carotenoids from microalgae. However, much more expensive photobioreactor systems and more strictly controlled sterile processes are needed to avoid contamination by heterotrophic organisms during mixotrophic cultivation processes, which makes mixotrophy, in fact, much more expensive than phototrophic cultivation. Recently technical breakthroughs have been reported to overcome contamination problems in photobioreactor systems traditionally used for phototrophic cultivation, and a much lower process cost of mixotrophic cultivation than that of phototrophic cultivation might be achieved for carotenoid production. These reviews intend to summarize recent technical advancements in mixotrophic cultivation of microalgae, to evaluate the economic viability of carotenoid production from mixotrophically cultivated microalgae, and to prospect mixotrophy as a strategy to produce a variety of carotenoids for industrial applications.

Keywords: microalgae, carotenoid, mixotrophy, biotechnology

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Authors: A. Jéglot, F. Plauborg, M. K. Schnorr, R. S. Sørensen, L. Elsgaard

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Artificial wetlands such as woodchip bioreactors are efficient tools to remove nitrate from agricultural wastewater with a minimized environmental impact. However, the temperature dependence of the microbiological nitrate removal prevents the woodchip bioreactors from being an efficient system when the water temperature drops below 8℃. To quantify and describe the temperature effects on nitrate removal efficiency, we studied nitrate-reducing enrichments from a woodchip bioreactor in Denmark based on samples collected in Spring and Fall. Growth was quantified as optical density, and nitrate and nitrous oxide concentrations were measured in time-course experiments to compare the growth of the microbial population and the nitrate conversion efficiencies at different temperatures. Ammonia was measured to indicate the importance of dissimilatory nitrate reduction to ammonia (DNRA) in nitrate conversion for the given denitrifying community. The temperature responses observed followed the increasing trend proposed by the Arrhenius equation, indicating higher nitrate removal efficiencies at higher temperatures. However, the growth and the nitrous oxide production observed at low temperature provided evidence of the psychrotolerance of the microbial community under study. The assays conducted showed higher nitrate removal from the microbial community extracted from the woodchip bioreactor at the cold season compared to the ones extracted during the warmer season. This indicated the ability of the bacterial populations in the bioreactor to evolve and adapt to different seasonal temperatures.

Keywords: agricultural waste water treatment, artificial wetland, denitrification, psychrophilic conditions

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Authors: Benaliouche Hana, Abdessemed Djamal, Meniai Abdessalem, Lesage Geoffroy, Heran Marc

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This paper presents a dynamic mathematical model of activated sludge which is able to predict the formation and degradation kinetics of SMP (Soluble microbial products) in membrane bioreactor systems. The model is based on a calibrated version of ASM1 with the theory of production and degradation of SMP. The model was calibrated on the experimental data from MBR (Mathematical modeling Membrane bioreactor) pilot plant. Analytical expressions have been developed, describing the concentrations of the main state variables present in the sludge matrix, with the inclusion of only six additional linear differential equations. The objective is to present a new dynamic mathematical model of activated sludge capable of predicting the formation and degradation kinetics of SMP (UAP and BAP) from the submerged membrane bioreactor (BRMI), operating at low organic load (C / N = 3.5), for two sludge retention times (SRT) fixed at 40 days and 60 days, to study their impact on membrane fouling, The modeling study was carried out under the steady-state condition. Analytical expressions were then validated by comparing their results with those obtained by simulations using GPS-X-Hydromantis software. These equations made it possible, by means of modeling approaches (ASM1), to identify the operating and kinetic parameters and help to predict membrane fouling.

Keywords: Activated Sludge Model No. 1 (ASM1), mathematical modeling membrane bioreactor, soluble microbial products, UAP, BAP, Modeling SMP, MBR, heterotrophic biomass

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Authors: Patrick Mueller, Alan Martin, Justin Stockwell, Robert Goldblatt

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Elevated concentrations of inorganic nitrogen (N) compounds (nitrate, nitrite, and ammonia) are a ubiquitous feature to mine-influenced drainages due to the leaching of blasting residues and use of cyanide in the milling of gold ores. For many mines, the management of N is a focus for environmental protection, therefore understanding the factors controlling the speciation and behavior of N is central to effective decision making. In this paper, the passive attenuation of ammonia and nitrite is described for three northern water bodies (two lakes and a tailings pond) influenced by mining activities. In two of the water bodies, inorganic N compounds originate from explosives residues in mine water and waste rock. The third water body is a decommissioned tailings impoundment, with N compounds largely originating from the breakdown of cyanide compounds used in the processing of gold ores. Empirical observations from water quality monitoring indicate nitrification (the oxidation of ammonia to nitrate) occurs in all three waterbodies, where enrichment of nitrate occurs commensurately with ammonia depletion. The N species conversions in these systems occurred more rapidly than chemical oxidation kinetics permit, indicating that microbial mediated conversion was occurring, despite the cool water temperatures. While nitrification of ammonia and nitrite to nitrate was the primary process, in all three waterbodies nitrite was consistently present at approximately 0.5 to 2.0 % of total N, even following ammonia depletion. The persistence of trace amounts of nitrite under these conditions suggests the co-occurrence denitrification processes in the water column and/or underlying substrates. The implications for N management in mine waters are discussed.

Keywords: explosives, mining, nitrification, water

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Authors: M. Nita-Lazar, E. Manea, C. Bumbac, A. Banciu, C. Stoica

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The population and economic growth have generated a significant new number of pollutant compounds which have to be degraded before reaching the environment. The wastewater treatment plants (WWTPs) have been the last barrier between the domestic and/or industrial wastewaters and the environment. At present, the conventional WWTPs have very high operational costs, most of them linked to the aeration process (60-65% from total energy costs related to wastewater treatment). In addition, they have had a low efficiency in pollutants removal such as pharmaceutical and other resilient anthropogenic compounds. In our study, we have been focused on new wastewater treatment strategies to enhance the efficiency of pollutants removal and decrease the wastewater treatment operational costs. The usage of mixed microalgae-bacteria granules technology generated high efficiency and low costs by a better harvesting and less expensive aeration. The intertrophic relationships between microalgae and bacteria have been characterized by the structure of the population community to their metabolic relationships. The results, obtained by microscopic studies, showed well-organized and stratified microalgae-bacteria granules where bacteria have been enveloped in the microalgal structures. Moreover, their population community structure has been modulated as well as their nitrification, denitrification processes (analysis based on qPCR genes expression) by the type of the pollutant compounds and amounts. In conclusion, the understanding and modulation of intertrophic relationships between microalgae and bacteria could be an economical and technological viable alternative to the conventional wastewater treatment. Acknowledgements: This research was supported by grant PN-III-P4-ID-PCE-2016-0865 from the Romanian National Authority for Scientific Research and Innovation CNCS/CCCDI-UEFISCDI.

Keywords: activated sludge, bacteria, granules, microalgae

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Authors: Qiongpeng Dan, Xiyao Li, Qiong Zhang, Yongzhen Peng

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The nutrients removal from wastewater is of great significance for global wastewater recycling and sustainable reuse. Traditional nitrogen and phosphorus removal processes are very dependent on the input of aeration and carbon sources, which makes it difficult to meet the low-carbon goal of energy saving and emission reduction. This study reported a proof-of-concept demonstration of integrating anammox and enhanced biological phosphorus removal (EBPR) by flocs management in a single-stage hybrid bioreactor (biofilms and flocs) for simultaneous nitrogen and phosphorus removal (SNPR). Excellent removal efficiencies of nitrogen (97.7±1.3%) and phosphorus (97.4±0.7%) were obtained in low C/N ratio (3.0±0.5) municipal wastewater treatment. Interestingly, with the loss of flocs, anammox bacteria (Ca. Brocadia) was highly enriched in biofilms, with relative and absolute abundances reaching up to 12.5% and 8.3×1010 copies/g dry sludge, respectively. The anammox contribution to nitrogen removal also rose from 32.6±9.8% to 53.4±4.2%. Endogenous denitrification by flocs was proven to be the main contributor to both nitrite and nitrate reduction, and flocs loss significantly promoted nitrite flow towards anammox, facilitating AnAOB enrichment. Moreover, controlling the floc's solid retention time at around 8 days could maintain a low poly-phosphorus level of 0.02±0.001 mg P/mg VSS in the flocs, effectively addressing the additional phosphorus removal burden imposed by the enrichment of phosphorus-accumulating organisms in biofilms. This study provides an update on developing a simple and feasible strategy for integrating anammox and EBPR for SNPR in mainstream municipal wastewater.

Keywords: anammox process, enhanced biological phosphorus removal, municipal wastewater, sustainable nutrients removal

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Authors: Keisuke Konishi, Yoshinari Hiroshiro, Kento Terashima, Atsushi Tsutsumi

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Imazu Bay plays an important role for endangered species such as horseshoe crabs and black-faced spoonbills that stay in the bay for spawning or the passing of winter. However, this bay is semi-enclosed with slow water exchange, which could lead to eutrophication under the condition of excess nutrient inflow to the bay. Therefore, quantification of nutrient inflow is of great importance. Generally, analysis of nutrient inflow to the bays takes into consideration nutrient inflow from only the river, but that from groundwater should not be ignored for more accurate results. The main objective of this study is to estimate the amounts of nutrient inflow from river and groundwater to Imazu Bay by analyzing water budget in Zuibaiji River Basin and loads of T-N, T-P, NO3-N and NH4-N. The water budget computation in the basin is performed using groundwater recharge model and quasi three-dimensional two-phase groundwater flow model, and the multiplication of the measured amount of nutrient inflow with the computed discharge gives the total amount of nutrient inflow to the bay. In addition, in order to evaluate nutrient inflow to the bay, the result is compared with nutrient inflow from geologically similar river basins. The result shows that the discharge is 3.50×107 m3/year from the river and 1.04×107 m3/year from groundwater. The submarine groundwater discharge accounts for approximately 23 % of the total discharge, which is large compared to the other river basins. It is also revealed that the total nutrient inflow is not particularly large. The sum of NO3-N and NH4-N loadings from groundwater is less than 10 % of that from the river because of denitrification in groundwater. The Shin Seibu Sewage Treatment Plant located below the observation points discharges treated water of 15,400 m3/day and plans to increase it. However, the loads of T-N and T-P from the treatment plant are 3.9 mg/L and 0.19 mg/L, so that it does not contribute a lot to eutrophication.

Keywords: Eutrophication, groundwater recharge model, nutrient inflow, quasi three-dimensional two-phase groundwater flow model, submarine groundwater discharge

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Authors: George Obinna Akuaka

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Solid waste disposal techniques in Nigeria pose an environmental threat to the environment and to nearby resident. The presence of microbial physical and chemical concentration in boreholes samples nearby dumpsite implies that groundwater is normally contaminated by leachate infiltration from an open landfill dumpsite. In this study, the physicochemical and microbial analyses of water samples from hand dug well in the site and boreholes were carried out around the active landfill and from different distances (50 m to 200 m). leachate samples collected were used to ascertain the effect or extent of contamination on the groundwater quality. A total of 5 leachate samples and 5 samples of groundwater were collected, and all samples were analyzed for various physical and chemical parameters according to the standard methods. These include pH, Electrical conductivity, Total dissolved solid, BOD, OD, Temperature, major cations such as Mg²+ Ca²+, Fe²+ Cu²+, major anions NO³-, Cl-,SO⁴- PO⁴-, Zn, Ar, Cd, Cr, Hg, Pb, Ni are the heavy metals and metalloids. The mean values of the physical and chemical parameters obtained from both sites were compared with the established of the World Health Organization (WHO). The leachate samples were found to be higher in the concentration of the results obtained than that of the boreholes water, and the recorded mean values of heavy metals were above approved standard minimum limits. The results indicated that mercury and copper were not found in all the borehole water samples. Microbial analyses showed that total heterotrophic bacteria mean count ranged from 10.6 X10⁷ cfu/ml to 2.04x10⁷cfu/ml and 9.5 X 10⁷ cfu/ml to 18.9 X 10⁷ cfu/ml in leachate and borehole samples respectively. It also revealed that almost at the bacteria isolated in the leachate were also found in the water samples. This results indicated that heavy pollution in all the samples with most physicochemical parameters and microbes showed traceable pollution, which occurred as a result of leachate infiltration into the ground water.

Keywords: physicochemical, landfill dumpsite, microbial, leachate, groundwater

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Authors: Nabil Elkhatri, Mohamed Louay Metougui, Ngonidzashe Chirinda

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Mining activities have left a legacy of degraded landscapes, prompting urgent efforts for ecological restoration. Reforestation holds promise as a potent tool to rehabilitate these old mining sites, with the potential to sequester carbon and contribute to climate change mitigation. This study focuses on evaluating the carbon stock and sequestration potential of reforestation species in the context of Morocco's mining areas, employing the DeNitrification-DeComposition (DNDC) model. The research is grounded in recognizing the need to connect theoretical models with practical implementation, ensuring that reforestation efforts are informed by accurate and context-specific data. Field data collection encompasses growth patterns, biomass accumulation, and carbon sequestration rates, establishing an empirical foundation for the study's analyses. By integrating the collected data with the DNDC model, the study aims to provide a comprehensive understanding of carbon dynamics within reforested ecosystems on old mining sites. The major findings reveal varying sequestration rates among different reforestation species, indicating the potential for species-specific optimization of reforestation strategies to enhance carbon capture. This research's significance lies in its potential to contribute to sustainable land management practices and climate change mitigation strategies. By quantifying the carbon stock and sequestration potential of reforestation species, the study serves as a valuable resource for policymakers, land managers, and practitioners involved in ecological restoration and carbon management. Ultimately, the study aligns with global objectives to rejuvenate degraded landscapes while addressing pressing climate challenges.

Keywords: carbon stock, carbon sequestration, DNDC model, ecological restoration, mining sites, Morocco, reforestation, sustainable land management.

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Authors: Keerthi Katam, Abhinav B. Tirunaghari, Vinod Vadithya, Toshiyuki Shimizu, Satoshi Soda, Debraj Bhattacharyya

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Researchers worldwide are increasingly focusing on the removal of carbon and nutrient from wastewater using algal-bacterial hybrid systems. Algae produce oxygen during photosynthesis, which is taken up by heterotrophic bacteria for mineralizing organic carbon to carbon dioxide. This phenomenon reduces the net mechanical aeration requirement of aerobic biological wastewater treatment processes. Consequently, the treatment cost is also reduced. Microalgae also participate in the treatment process by taking up nutrient (N, P) from wastewater. Algal biomass, if harvested, can generate value-added by-products. The aim of the present study was to compare the performance of two systems - System A (mixed microalgae and bacteria) and System B (diatoms and bacteria) in treating kitchen wastewater (KWW). The test reactors were operated at five different solid retention times (SRTs) -2, 4, 6, 8, and 10-days in draw-and-fill mode. The KWW was collected daily from the dining hall-kitchen area of the Indian Institute of Technology Hyderabad. The influent and effluent samples were analyzed for total organic carbon (TOC), total nitrogen (TN) using TOC-L analyzer. A colorimetric method was used to analyze anionic surfactant. Phosphorus (P) and chlorophyll were measured by following standard methods. The TOC, TN, and P of KWW were in the range of 113.5 to 740 mg/L, 2 to 22.8 mg/L, and 1 to 4.5 mg/L, respectively. Both the systems gave similar results with 85% of TOC removal and 60% of TN removal at 10-d SRT. However, the anionic surfactant removal in System A was 99% and 60% in System B. The chlorophyll concentration increased with an increase in SRT in both the systems. At 2-d SRT, no chlorophyll was observed in System B, whereas 0.5 mg/L was observed in System A. At 10-d SRT, the chlorophyll concentration in System A was 7.5 mg/L, whereas it was 4.5 mg/L in System B. Although both the systems showed similar performance in treatment, the increase in chlorophyll concentration suggests that System A demonstrated a better algal-bacterial symbiotic relationship in treating KWW than System B.

Keywords: diatoms, microalgae, retention time, wastewater treatment

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Authors: I. Kamika, S. Azizi, M. Tekere

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Nanotechnology promises significant improvements of advanced materials and manufacturing techniques with a vast range of applications, which are critical for the future competitiveness of national industries. The manipulations and productions of materials, whilst, controlling the optical properties and surface area to a nanosize scale enabled a birth of a new field known as nanotechnology. However, their rapidly developing industry raises concerns about the environmental impacts of nanoparticles, as their effects on functional bacterial community in wastewater treatment remain unclear. The present research assessed the impact of cerium Oxide nanoparticles (nCeO) on the bacterial microbiome of an activated sludge system, which influenced its performance of this system on nutrient removal. Out of 15875 reads sequenced, a total of 13133 reads were non-chimeric. The wastewater samples were more dominant to the unclassified bacteria (51.07% of bacteria community) followed with the classified bacteria (48.93). Proteobacteria was the most dominant phylum in both classified and unclassified bacteria, whereas 18% of bacteria could even not be assigned a phylum and remained unclassified suggesting hitherto vast untapped microbial diversity. The bacterial operational taxonomic units (OTUs) ranged from 1014 to 2629 over the experimental period. The denitrification related species including Diaphorobacter species, Thauera species and those in the Sphaerotilus and Leptothrix group were found to be inhibited in a high concentration of CeO-NP. The diversity indices suggested that the bacterial community inhabiting the wastewater samples were less diverse as the concentration of CeO increases. The canonical correspondence analysis (CCA) results highlighted that the bacterial community variance had the strongest relationship with water temperature, conductivity, pH, and dissolved oxygen (DO) content as well as nCeO. The results provided the relationships between the microbial community and environmental variables in the wastewater samples.

Keywords: bacterial community, next generation, cerium oxide, wastewater, activated sludge, nanoparticles, nanotechnology

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Authors: Namratha Pai, K. Vasantharaj, K. Haribabu

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Our Team is developing an innovative cost effective biological technique to desulfurize crude oil. ’Sulphur’ is found to be present in crude oil samples from .05% - 13.95% and its elimination by industrial methods is expensive currently. Materials required :- Alicyclobacillus acidoterrestrius, potato dextrose agar, oxygen, Pyragallol and inert gas(nitrogen). Method adapted and proposed:- 1) Growth of bacteria studied, energy needs. 2) Compatibility with crude-oil. 3) Reaction rate of bacteria studied and optimized. 4) Reaction development by computer simulation. 5) Simulated work tested by building the reactor. The method being developed requires the use of bacteria Alicyclobacillus acidoterrestrius - an acidothermophilic heterotrophic, soil dwelling aerobic, Sulfur bacteria. The bacteria are fed to crude oil in a unique manner. Its coated onto potato dextrose agar beads, cultured for 24 hours (growth time coincides with time when it begins reacting) and fed into the reactor. The beads are to be replenished with O2 by passing them through a jacket around the reactor which has O2 supply. The O2 can’t be supplied directly as crude oil is inflammable, hence the process. Beads are made to move around based on the concept of fluidized bed reactor. By controlling the velocity of inert gas pumped , the beads are made to settle down when exhausted of O2. It is recycled through the jacket where O2 is re-fed and beads which were inside the ring substitute the exhausted ones. Crude-oil is maintained between 1 atm-270 M Pa pressure and 45°C treated with tartaric acid (Ph reason for bacteria growth) for optimum output. Bacteria being of oxidising type react with Sulphur in crude-oil and liberate out SO4^2- and no gas. SO4^2- is absorbed into H2O. NaOH is fed once reaction is complete and beads separated. Crude-oil is thus separated of SO4^2-, thereby Sulphur, tartaric acid and other acids which are separated out. Bio-corrosion is taken care of by internal wall painting (phenolepoxy paints). Earlier methods used included use of Pseudomonas and Rhodococcus species. They were found to be inefficient, time and energy consuming and reduce the fuel value as they fed on skeleton.

Keywords: alicyclobacillus acidoterrestrius, potato dextrose agar, fluidized bed reactor principle, reaction time for bacteria, compatibility with crude oil

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Authors: Mary A. Bisi-Johnson, Kehinde A. Adediran, Saheed A. Akinola, Hamzat A. Oyelade

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Some of the main problems man contends with are the quantity (source and amount) and quality of water in Nigeria. Scarcity leads to water being obtained from various sources and microbiological contaminations of the water may thus occur between the collection point and the point of usage. Thus, this study aims to assess the general and microbiological quality of domestic water sources and household stored water used within selected areas in Ile-Ife, South-Western part of Nigeria for microbial contaminants. Physicochemical and microbiological examination were carried out on 45 source and stored water samples collected from well and spring in three different local government areas i.e. Ife east, Ife-south, and Ife-north. Physicochemical analysis included pH value, temperature, total dissolved solid, dissolved oxygen, and biochemical oxygen demand. Microbiology involved most probable number analysis, total coliform, heterotrophic plate, faecal coliform, and streptococcus count. The result of the physicochemical analysis of samples showed anomalies compared to acceptable standards with the pH value of 7.20-8.60 for stored and 6.50-7.80 for source samples as the total dissolved solids (TDS of stored 20-70mg/L, source 352-691mg/L), dissolved oxygen (DO of stored 1.60-9.60mg/L, source 1.60-4.80mg/L), biochemical oxygen demand (BOD stored 0.80-3.60mg/L, source 0.60-5.40mg/L). General microbiological quality indicated that both stored and source samples with the exception of a sample were not within acceptable range as indicated by analysis of the MPN/100ml which ranges (stored 290-1100mg/L, source 9-1100mg/L). Apart from high counts, most samples did not meet the World Health Organization standard for drinking water with the presence of some pathogenic bacteria and fungi such as Salmonella and Aspergillus spp. To annul these constraints, standard treatment methods should be adopted to make water free from contaminants. This will help identify common and likely water related infection origin within the communities and thus help guide in terms of interventions required to prevent the general populace from such infections.

Keywords: domestic, microbiology, physicochemical, quality, water

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Authors: Eva Slaninova, Diana Cernayova, Zuzana Sedrlova, Katerina Mrazova, Petr Sedlacek, Jana Nebesarova, Stanislav Obruca

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Cyanobacteria are gram-negative prokaryotes belonging to a group of photosynthetic bacteria. In comparison with heterotrophic microorganisms, cyanobacteria utilize atmospheric nitrogen and carbon dioxide without any additional substrates. This ability of these microorganisms could be employed in biotechnology for the production of bioplastics, concretely polyhydroxyalkanoates (PHAs) which are primarily accumulated as a storage material in cells in the form of intracellular granules. In this study, there two cyanobacterial cultures from genera Synechocystis were used, namely Synechocystic sp. PCC 6803 and Synechocystis salina CCALA 192. There were optimized and used several various approaches, including microscopic techniques such as cryo-scanning electron microscopy (Cryo-SEM) and transmission electron microscopy (TEM), and fluorescence lifetime imaging microscopy using Nile red as a fluorescent probe (FLIM). Due to these instrumental techniques, the morphology of intracellular space and surface of cells were characterized. The next group of methods which were employed was spectroscopic techniques such as UV-Vis spectroscopy measured in two modes (turbidimetry and integration sphere) and Fourier transform infrared spectroscopy (FTIR). All these diverse techniques were used for the detection and characterization of pigments (chlorophylls, carotenoids, phycocyanin, etc.) and PHAs, in our case poly (3-hydroxybutyrate) (P3HB). To verify results, gas chromatography (GC) was employed concretely for the determination of the amount of P3HB in biomass. Cyanobacteria were also characterized as polyhydroxybutyrate producers by flow cytometer, which could count cells and at the same time distinguish cells including P3HB and without due to fluorescent probe called BODIPY and live/dead fluorescent probe SYTO Blue. Based on results, P3HB content in cyanobacteria cells was determined, as also the overall fitness of the cells. Acknowledgment: Funding: This study was partly funded by the projectGA19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), project I 4082-B25.

Keywords: cyanobacteria, fluorescent probe, microscopic techniques, poly(3hydroxybutyrate), spectroscopy, chromatography

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