Biotechnological product potential of Auxenochlorella protothecoides including biologically active compounds (BACs) under nitrogen stress conditions.

Nitrogen stress can influence microalgae's growth characteristics, and microalgae grown in nitrogen-deficient conditions may produce higher or lower levels of biotechnological products as a result of metabolic changes. In photoautotrophic and heterotrophic cultures, nitrogen limitation has been proven effective in promoting lipid accumulation. In spite of this, no study has demonstrated a significant correlation between lipid content and other biotechnological products such as bioactive compounds (BACs). This research examines a strategy for lipid accumulation as well as the potential production of BACs with antibacterial properties in parallel with that strategy. This concept involved the treatment of the microalga Auxenochlorella protothecoides with low and high concentrations of ammonium (NH4+). This particular experiment reached a maximum lipid content of 59.5% using a 0.8 mM NH4+ concentration, resulting in the yellowing of the chlorophyll levels. Agar diffusion assays were conducted to determine the antibacterial activity of different extracts derived from the biomass when stressed with different levels of nitrogen. Algal extracts prepared by a variety of solvents showed different levels of antibacterial activity against representative strains of both gram-negative (Escherichia coli) and gram-positive (Staphylococcus aureus) bacteria. Among the extracts tested, 500 mg/L ethyl acetate extract had the greatest antibacterial activity against Escherichia coli. In order to identify the components responsible for the extract's antibacterial activity, fatty acid methyl ester (FAME) analysis was performed. It has been suggested that the lipid fraction may be a valuable indicator of these activities since some lipid components are known to possess antimicrobial properties. In this regard, it was found that the amount of polyunsaturated fatty acid (PUFA) significantly decreased by 53.4% under the conditions with the highest antibacterial activity observed.

Author Correction: Effect of iron and magnesium addition on population dynamics and high value product of microalgae grown in anaerobic liquid digestate.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Effect of iron and magnesium addition on population dynamics and high value product of microalgae grown in anaerobic liquid digestate.

In this study, FeSO4 supplementation ranging from 0 to 4.5 mM, and MgSO4 supplementation ranging from 0 to 5.1 mM were investigated to observe the effect on the population dynamics, biochemical composition and fatty acid content of mixed microalgae grown in Anaerobic Liquid Digestate (ALD). Overall, 3.1 mM FeSO4 addition into ALD increased the total protein content 60% and led to highest biomass (1.56 g L-1) and chlorophyll-a amount (18.7 mg L-1) produced. Meanwhile, 0.4 mM MgSO4 addition increased the total carotenoid amount 2.2 folds and slightly increased the biomass amount. According to the microbial community analysis, Diphylleia rotans, Synechocystis PCC-6803 and Chlorella sorokiniana were identified as mostly detected species after confirmation with 4 different markers. The abundance of Chlorella sorokiniana and Synechocystis PCC-6803 increased almost 2 folds both in iron and magnesium addition. On the other hand, the dominancy of Diphylleia rotans was not affected by iron addition while drastically decreased (95%) with magnesium addition. This study helps to understand how the dynamics of symbiotic life changes if macro elements are added to the ALD and reveal that microalgae can adapt to adverse environmental conditions by fostering the diversity with a positive effect on high value product.

Impact of ultrasonic pre-treatment on domestic sludge digestion performance and microbial community dynamics.

Ultrasonication-assisted sludge digestion technology is a lately used alternative sludge treatment method in wastewater treatment plants (WWTPs). This study focused on determining the influence of ultrasonication on aerobic and anaerobic sludge digestion, two most commonly used sludge handling processes, as well as on the investigation of microbial community structure after digestion. The effect of ultrasonication as a pre-treatment technique prior to sludge digestion on microbial population dynamics was not yet investigated comprehensively. Sludge sample taken from the primary and secondary settling tanks of a domestic wastewater treatment plant was used during the experiments. Based on the relevant data, while applied ultrasonication did not improve the anaerobic digestion efficiency, progress was achieved in the sludge dewaterability characteristics at the end of aerobic digestion. According to the results of both denaturant gradient gel electrophoresis and pyrosequencing data, ultrasonic pre-treatment decreased the richness of the microbial population in aerobic digestion, whereas increased the biocomplexity of the population in anaerobic digestion. We revealed that sludge pre-treatment with ultrasonication does not always improve the digestion performance. Composition of the sludge was the main factor defining the digestion performance.

Impact of salinity on the population dynamics of microorganisms in a membrane bioreactor treating produced water.

Biomass characteristics may change subject to a salinity increase when treating high strength wastewater. In this study, the impact of salinity in a membrane bioreactor (MBR) was investigated for the treatment of produced water (PW). MBR was operated as a pre-treatment prior to nanofiltration (NF) and reverse osmosis (RO). Mixed PW, that was originated from different oil, gas, and oil-gas wells, were subjected to pre-treatment for 146 days including three different operational phases: Low-salinity (~10 mS/cm), gradual increased salinity (10-40 mS/cm) and high salinity (~40 mS/cm). The results of this study showed that microorganisms could adapt using real wastewater and treat PW up to a certain level of the salinity. At high salinity levels of PW, the floc structure started to be disrupted and membrane fouling was accelerated. PCR-DGGE analysis confirmed the changes in microbial communities' composition in relation with high salinity. The results of the final treatment experiments presented that NF and RO treatment produced high-quality effluents that could be suitable for reuse.

Co-digestion performance of organic fraction of municipal solid waste with leachate: Preliminary studies.

The main aim of the study was to evaluate the co-digestion performance of OFMSW with different wastes. Leachate, reverse osmosis (RO) concentrate collected from a leachate treatment facility and dewatered sewage sludge taken from a wastewater treatment plant (WWTP) were used for co-digestion in this paper. An extra effort was made to observe the effect of leachate inclusion in the co-digestion. In the study, the mono-digestion of OFMSW, leachate, RO concentrate and sewage sludge as well as digestion of 7 different waste mixtures were carried out for this objective. The experiments were carried out for approximately 50days under mesophilic conditions. The highest methane yield was 785L CH4/kg VSadded in the reactor, which had only OFMSW. While the methane yield derived from OFMSW was found higher than previous studies, methane yield of leachate was found to be 110L CH4/kg VSadded, which was lower than findings in the literature. The mono-substrate of OFMSW was followed by the reactor of having waste mixture of leachate+sewage sludge+OFMSW+water (C7) with 391L CH4/kg VSadded, which was the only combination included water. In order to understand the effect of leachate and water inclusions on co-digestion, two separate waste combinations; leachate+sewage sludge+OFMSW+water (C7) and leachate+sewage sludge+OFMSW (C1) were prepared that had different amounts of leachate but same amounts of other wastes. The methane yield of leachate+sewage sludge+OFMSW+water (C7) indicated that addition of some water instead of leachate could stimulate biogas production. Methane yield of this reactor was found to be 71% higher than the waste combination of leachate+sewage sludge+OFMSW (C1). It could be thought that the high amount of non-biodegradable matters in leachate could be responsible for lower methane yield in leachate+sewage sludge+OFMSW (C1) reactor. Methane yields of the reactors showed that co-digestion of OFMSW and leachate could be a solution not only for treatment of leachate and but also increasing the biogas potential of leachate. Leachate addition could also adjust optimum total solids (TS) content in anaerobic digestion. It was also understood that RO concentrate did not affect the methane yield in a negative way. The similar characterization of leachate and RO concentrate in this study could offer the utilization of RO concentrate instead of leachate. The findings showed that volatile solids (VS) removals were changed from 32% to 61% in the reactors. While the reactor of leachate+RO concentrate+OFMSW (C6) had the highest VS removal, the reactor of the sole substrate leachate had the lowest VS removal.

Desulfatirhabdium butyrativorans gen. nov., sp. nov., a butyrate-oxidizing, sulfate-reducing bacterium isolated from an anaerobic bioreactor.

A novel sulfate-reducing bacterium, strain HB1(T), was isolated from an upflow anaerobic sludge blanket (UASB) reactor treating paper-mill wastewater operated at 37 degrees C. Cells of strain HB1(T) were oval to rod-shaped, 1-1.3 microm wide and 2.6-3.5 microm long and Gram-negative. The optimum temperature for growth was 28-30 degrees C. In the presence of sulfate, the isolate was able to grow on H(2)/acetate, formate, ethanol, propionate, fumarate, succinate, butyrate, crotonate, catechol, benzoate, 4-hydroxybenzoate, palmitate and stearate. The isolate only grew on H(2) when acetate was added as a carbon source; when grown on formate, acetate was not required. Growth was also possible on pyruvate and crotonate without an electron acceptor. The isolate showed very poor growth on acetate. Thiosulfate and sulfate were used as electron acceptors. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain HB1(T) represents a novel lineage within the Deltaproteobacteria; sequence similarities between strain HB1(T) and members of other related genera were less than 91%. Strain HB1(T) was also distinguished from members of related genera based on differences in several phenotypic characteristics. It is a member of the family Desulfobacteraceae. The major cellular fatty acids of strain HB1(T) were C(16:0), iso-C(15:0), anteiso-C(15:0) and C(14:0). beta-Hydroxy fatty acids were also present in the range of C(14:0) to C(18:0), of which C(16:0) was the most abundant. The G+C content of the DNA was 55.1 mol%. Based on physiological, biochemical and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence analysis, strain HB1(T) is considered to represent a novel species in a new genus, for which the name Desulfatirhabdium butyrativorans gen. nov., sp. nov. is proposed. The type strain of Desulfatirhabdium butyrativorans is HB1(T) (=DSM 18734(T) =JCM 14470(T)).

Molecular characterization of mesophilic and thermophilic sulfate reducing microbial communities in expanded granular sludge bed (EGSB) reactors.

The microbial communities established in mesophilic and thermophilic expanded granular sludge bed reactors operated with sulfate as the electron acceptor were analyzed using 16S rRNA targeted molecular methods, including denaturing gradient gel electrophoresis, cloning, and phylogenetic analysis. Bacterial and archaeal communities were examined over 450 days of operation treating ethanol (thermophilic reactor) or ethanol and later a simulated semiconductor manufacturing wastewater containing citrate, isopropanol, and polyethylene glycol 300 (mesophilic reactor), with and without the addition of copper(II). Analysis, of PCR-amplified 16S rRNA gene fragments using denaturing gradient gel electrophoresis revealed a defined shift in microbial diversity in both reactors following a change in substrate composition (mesophilic reactor) and in temperature of operation from 30 degrees C to 55 degrees C (thermophilic reactor). The addition of copper(II) to the influent of both reactors did not noticeably affect the composition of the bacterial or archaeal communities, which is in agreement with the very low soluble copper concentrations (3-310 microg l(-1)) present in the reactor contents as a consequence of extensive precipitation of copper with biogenic sulfides. Furthermore, clone library analysis confirmed the phylogenetic diversity of sulfate-reducing consortia in mesophilic and thermophilic sulfidogenic reactors operated with simple substrates.

Enrichment and detection of microorganisms involved in direct and indirect methanogenesis from methanol in an anaerobic thermophilic bioreactor.

To gain insight into the microorganisms involved in direct and indirect methane formation from methanol in a laboratory-scale thermophilic (55 degrees C) methanogenic bioreactor, reactor sludge was disrupted and serial dilutions were incubated in specific growth media containing methanol and possible intermediates of methanol degradation as substrates. With methanol, growth was observed up to a dilution of 10(8). However, when Methanothermobacter thermoautotrophicus strain Z245 was added for H2 removal, growth was observed up to a 10(10)-fold dilution. With H2/CO2 and acetate, growth was observed up to dilutions of 10(9) and 10(4), respectively. Dominant microorganisms in the different dilutions were identified by 16S rRNA-gene diversity and sequence analysis. Furthermore, dilution polymerase chain reaction (PCR) revealed a similar relative abundance of Archaea and Bacteria in all investigated samples, except in enrichment with acetate, which contained 100 times less archaeal DNA than bacterial DNA. The most abundant bacteria in the culture with methanol and strain Z245 were most closely related to Moorella glycerini. Thermodesulfovibrio relatives were found with high sequence similarity in the H2/CO2 enrichment, but also in the original laboratory-scale bioreactor sludge. Methanothermobacter thermoautotrophicus strains were the most abundant hydrogenotrophic archaea in the H2/CO2 enrichment. The dominant methanol-utilizing methanogen, which was present in the 10(8)-dilution, was most closely related to Methanomethylovorans hollandica. Compared to direct methanogenesis, results of this study indicate that syntrophic, interspecies hydrogen transfer-dependent methanol conversion is equally important in the thermophilic bioreactor, confirming previous findings with labeled substrates and specific inhibitors.

Advanced oxidation of biologically pretreated Baker's Yeast Industry effluents for high recalcitrant COD and color removal.

The aim of this study was to investigate the effectiveness of chemical oxidation by applying ozonation, ozonation with hydrogen peroxide and Fenton's processes for decolorization and residual COD removal of biologically pretreated baker's yeast industry (BYI) effluents. Baker's yeast industry effluents characterizing with high COD, TKN, dark color, and non-biodegradable organic pollutants. The batch tests were performed to determine the optimum operating conditions including pH, O3, H2O2, and FeSO4 dosages, molar ratio of Fe2+/H2O2 and reaction time. It was noticed that H2O2 significantly reduced the reaction times for the same ozone dosages: however, COD and color removals were not remarkable. In the Fenton's oxidation studies, the removal efficiencies of COD and color for 30 min reaction time for three different types of BYI effluents were found about 86 and 92%, respectively. Experimental results of the presented study have clearly indicated that the Fenton's oxidation technology is capable to fate almost all parts of the organics which consist of both soluble initial and microbial inert fractions of COD for baker's yeast effluents. Effluents from the Fenton's oxidation process can satisfy effluent standards for COD and color in general.

Anaerobic digestion of municipal sludges with high silt content using granular seed.

The performance of anaerobic digestion of municipal sludge having high inorganic solid/silt in Tuzla Wastewater Treatment Plant (TWWTP) in Istanbul was evaluated using granular seed. High silt/solid content is the main problem related to wastewater collection system in TWWTP. Due to high TSS concentration of the influent, inorganic solid ratio reaches to 60-70% in the primary sludge that makes sludge digestion very difficult. The aim of this study was to evaluate the detrimental effects of inorganic silt/clay on anaerobic sludge solubilization of the primary sludge (PS), of the biological sludge (BS), and of the mixed sludge (50%PS + 50%BS). Anaerobic solubilization was carried out in continuously stirred anaerobic reactors at mesophilic temperature (35 degrees C). Results indicated significant total biogas productions in all reactors during digestion period. The biogas production of the primary sludge was higher than the biogas productions of the mixed sludge and the biological sludge. The methane contents of the primary, biological. and mixed sludges were found as around 72, 78, and 75%, respectively. When all types of sludges were used as substrates, the reactors removed VSS with a corresponding production of Soluble COD (SCOD). The greatest degree of hydrolysis/acidogenesis was observed in all types of sludges in about 10 days of operation and became almost stable which indicated that the reactors were still in the hydrolysis/acidogenesis phase. VFAs measurements were well matched with SCOD results. In all reactors, observation of the conversion into methanogenic phase in about 1.5 month was not possible which might be attributed to high inorganic silt/clay and salinity content of TWWTP. High inorganic silt/clay and salinity might have decreased the biodegradability of all sludges and methanogenesis could not operate in early periods of digestion. Higher SCOD decreases might have been possible if longer treatment time and more optimal conditions for anaerobic solubilization were applied. The pH did not fall below the unfavorable range in all reactors during the study.

Advanced physico-chemical treatment experiences on young municipal landfill leachates.

In this study, Membrane Filtration (UF+RO), Struvite (MAP) precipitation and ammonia stripping alternatives were studied on biologically pre-treated Landfill Leachate. The results indicated that the system including the Upflow Anaerobic Sludge Blanket Reactor (UASBR) and Membrane Reactors (UF+RO) has been offered as an appropriate treatment alternative for young landfill leachates. This system provided high removals of COD, colour and conductivity (>98-99%). For ammonia removal, struvite precipitation was applied at the stoichiometric ratio (Mg:NH4:PO4=1:1:1) to anaerobically pre-treated raw landfill leachate effluent having an influent ammonium concentration of 2240 mg/l. Maximum ammonium nitrogen removal was observed as 85% at pH of 9.2. In ammonia stripping following 2 h of aeration, the removal was 72% at pH=12 while the removals were around 20% at pH=10 and pH=11. When membrane reactor, and struvite precipitation or ammonia stripping was applied to anaerobically pre-treated effluents, the results indicated that each system could be used as an appropriate post-treatment option for young landfill leachates. In economic aspect, ammonia stripping was found as the cheapest alternative with high ammonium removal. However, when both high COD and ammonium removals were to be achieved membrane technology such as UF+RO (SW) could be considered as the most appropriate system due to the fact that COD removal could be obtained very low by ammonia stripping.