This document summarizes a study examining the effects of hydraulic retention time (HRT), carbon source, and nitrogen source on bacterial communities grown using the effluent from a recirculation aquaculture system as a substrate. The study found that HRT influenced the bacterial community composition, with a 2 hour HRT favoring Acinetobacter spp. and a 7 hour HRT favoring Rhizobium and Shinella spp. The carbon source also impacted the community, with molasses favoring Aquaspirillum serpens. Providing both ammonia and nitrate as nitrogen sources led to bacteria related to Sphaerotilus, Flavobacterium, and Jonesia genera. The results
This document is a research article that examines the effects of the pesticide carbofuran on the early life stages of Nile tilapia (Oreochromis niloticus). The article describes experiments exposing tilapia larvae to various concentrations of carbofuran to determine mortality rates, effects on cholinesterase enzyme activity, and impacts on behaviors like vision, swimming, prey capture, and response to predators. The results showed sublethal carbofuran exposure can inhibit cholinesterase activity, impair vision and reduce swimming speed, prey capture attempts, growth, and ability to avoid predators in tilapia larvae.
2017 - Comparison of nitrifying microbial communities of two full-scale membr...WALEBUBLÉ
Barbarroja, P., Moreno-Mesonero, L., Zornoza, A., Fernández-Navarro, J., Alonso, J.L., Muñagorri, F., García, C., Álvarez, C. (2017) Comparison of nitrifying microbial communities of two full-scale membrane bioreactors treating wastewaters from municipal solid wastes using 16S rDNA gene amplicon sequencing. 7th congress of European microbiologists FEMS 2017, Valencia, Spain, 9-13 July 2017.
2017 - Environmental ordination of nitrifying bacterial community dynamics in...WALEBUBLÉ
Biological nitrification-denitrification is commonly used for nitrogen removal in Wastewater Treatment Plants (WWTPs). Nitrification, is the sequential oxidation of ammonia via nitrite to nitrate. This process is catalysed by ammonia-oxidizing bacteria and archaea (AOB and AOA) and nitrite-oxidizing bacteria (NOB), whose cooperation is needed to achieve complete nitrification. They are a phylogenetically diverse guild with pronounced ecological niche specialization and they differ from each other in fundamental physiological and molecular traits. Although the nitrification process in WWTPs has been investigated in depth, the response of microbial
communities are still a focus of considerable interest due to their high sensitivity to inhibitory compounds and environmental factors, that results in repeated breakdowns of nitrification performance. Most of studies have been mainly descriptive and/or exploratory and environmental interpretation has not been addressed. In this study, we focus on the environmental ordination of the relationships between biological variables (nitrifying bacterial community) and physicochemical variables (nitrogen compounds and environmental conditions), to propose new strategies to improve the performance of the nitrogen removal process in WWTPs.
2019 - Profiling of filamentous bacteria in activated sludge by 16s RNA ampli...WALEBUBLÉ
Abstract: In this study, filamentous bacteria in the activated sludge of a WWTP were investigated throughout a one-year period using high-throughput short-read (Illumina) and full-length (PacBio) 16S rRNA gene amplicon sequencing. The results showed that a total of 28 filamentous bacteria genera
were identified using Illumina sequencing. Also, we found 25 species using PacBio sequencing, belonging to Curvibacter, Mycobacterium, Haliscomenobacter, Defluvicoccus, Sphaerotilus, Thiothrix, Leptothrix, Gordonia and Tetrasphaera genera. Active Volatile Suspended Solids (AVSS) were
calculated from ATP data contained in living microorganisms, this parameter represents the living biomass concentration, and the food/microorganisms ratio (F/M ratio) was calculated using AVSS instead of MLVSS. To assess the contribution of the F/M ratio to the variability observed in the filamentous bacteria structure we carried out distance-based linear models (DISTLM) and distancebased redundancy analysis (dbRDA).
This document summarizes a study evaluating the use of algae to convert manure nutrients into animal feed. The goals were to optimize algal growth, nutritional value, pathogen inactivation, and toxin control. Algae were grown in ponds fed with dairy lagoon effluent. Nutrient uptake, biomass composition, and dominant algal species were monitored over time. Several algal strains were isolated and characterized. The results support using algae to beneficially reuse manure while producing safe, nutritious animal feed. Future work will refine techniques for safety testing and pathogen identification.
2017 - Environmental Ordination of Filamentous Bacteria in Activated SludgeWALEBUBLÉ
Reference:
Zornoza, A., Serrano, S. and Alonso, J.L. (2017) Environmental Ordination of Filamentous Bacteria in Activated Sludge. In: Abstracts of the 7th congress of European microbiologists FEMS 2017, Valencia, Spain, 9-13 July 2017.
his study investigated the microbial community in a full scale anaerobic baffled reactor and sequencing batch reactor system for oil-produced water treatment in summer and winter. The community structures of fungi and bacteria were analyzed through polymerase chain reaction–denaturing gradient gel electrophoresis and Illumina high-throughput sequencing, respectively. Chemical oxygen demand effluent concentration achieved lower than 50 mg/L level after the system in both summer and winter, however, chemical oxygen demand removal rates after anaerobic baffled reactor treatment system were significant higher in summer than that in winter, which conformed to the microbial community diversity. Saccharomycotina, Fusarium, and Aspergillus were detected in both anaerobic baffled reactor and sequencing batch reactor during summer and winter. The fungal communities in anaerobic baffled reactor and sequencing batch reactor were shaped by seasons and treatment units, while there was no correlation between abundance of fungi and chemical oxygen demand removal rates. Compared to summer, the total amount of the dominant hydrocarbon degrading bacteria decreased by 10.2% in anaerobic baffled reactor, resulting in only around 23% of chemical oxygen demand was removed in winter. Although microbial community significantly varied in the three parallel sulfide reducing bacteria, the performance of these bioreactors had no significant difference between summer and winter.
2008 - Molecular microbial and chemical investigation of the bioremediation o...WALEBUBLÉ
The document describes a study that used laboratory-scale bioreactors to investigate the biodegradation of two-phase olive mill waste (TPOMW) by its indigenous microbiota under different conditions. The effects of nutrient addition and aeration on bioremediation and microbial community changes were evaluated. Analysis found that nutrient addition and aeration led to greater decreases in polyphenolic content and increases in the fungal to bacterial ratio. Molecular identification of bacteria and fungi in the bioreactors identified several genera present, with fungi like Penicillium and Candida dominant.
This document is a research article that examines the effects of the pesticide carbofuran on the early life stages of Nile tilapia (Oreochromis niloticus). The article describes experiments exposing tilapia larvae to various concentrations of carbofuran to determine mortality rates, effects on cholinesterase enzyme activity, and impacts on behaviors like vision, swimming, prey capture, and response to predators. The results showed sublethal carbofuran exposure can inhibit cholinesterase activity, impair vision and reduce swimming speed, prey capture attempts, growth, and ability to avoid predators in tilapia larvae.
2017 - Comparison of nitrifying microbial communities of two full-scale membr...WALEBUBLÉ
Barbarroja, P., Moreno-Mesonero, L., Zornoza, A., Fernández-Navarro, J., Alonso, J.L., Muñagorri, F., García, C., Álvarez, C. (2017) Comparison of nitrifying microbial communities of two full-scale membrane bioreactors treating wastewaters from municipal solid wastes using 16S rDNA gene amplicon sequencing. 7th congress of European microbiologists FEMS 2017, Valencia, Spain, 9-13 July 2017.
2017 - Environmental ordination of nitrifying bacterial community dynamics in...WALEBUBLÉ
Biological nitrification-denitrification is commonly used for nitrogen removal in Wastewater Treatment Plants (WWTPs). Nitrification, is the sequential oxidation of ammonia via nitrite to nitrate. This process is catalysed by ammonia-oxidizing bacteria and archaea (AOB and AOA) and nitrite-oxidizing bacteria (NOB), whose cooperation is needed to achieve complete nitrification. They are a phylogenetically diverse guild with pronounced ecological niche specialization and they differ from each other in fundamental physiological and molecular traits. Although the nitrification process in WWTPs has been investigated in depth, the response of microbial
communities are still a focus of considerable interest due to their high sensitivity to inhibitory compounds and environmental factors, that results in repeated breakdowns of nitrification performance. Most of studies have been mainly descriptive and/or exploratory and environmental interpretation has not been addressed. In this study, we focus on the environmental ordination of the relationships between biological variables (nitrifying bacterial community) and physicochemical variables (nitrogen compounds and environmental conditions), to propose new strategies to improve the performance of the nitrogen removal process in WWTPs.
2019 - Profiling of filamentous bacteria in activated sludge by 16s RNA ampli...WALEBUBLÉ
Abstract: In this study, filamentous bacteria in the activated sludge of a WWTP were investigated throughout a one-year period using high-throughput short-read (Illumina) and full-length (PacBio) 16S rRNA gene amplicon sequencing. The results showed that a total of 28 filamentous bacteria genera
were identified using Illumina sequencing. Also, we found 25 species using PacBio sequencing, belonging to Curvibacter, Mycobacterium, Haliscomenobacter, Defluvicoccus, Sphaerotilus, Thiothrix, Leptothrix, Gordonia and Tetrasphaera genera. Active Volatile Suspended Solids (AVSS) were
calculated from ATP data contained in living microorganisms, this parameter represents the living biomass concentration, and the food/microorganisms ratio (F/M ratio) was calculated using AVSS instead of MLVSS. To assess the contribution of the F/M ratio to the variability observed in the filamentous bacteria structure we carried out distance-based linear models (DISTLM) and distancebased redundancy analysis (dbRDA).
This document summarizes a study evaluating the use of algae to convert manure nutrients into animal feed. The goals were to optimize algal growth, nutritional value, pathogen inactivation, and toxin control. Algae were grown in ponds fed with dairy lagoon effluent. Nutrient uptake, biomass composition, and dominant algal species were monitored over time. Several algal strains were isolated and characterized. The results support using algae to beneficially reuse manure while producing safe, nutritious animal feed. Future work will refine techniques for safety testing and pathogen identification.
2017 - Environmental Ordination of Filamentous Bacteria in Activated SludgeWALEBUBLÉ
Reference:
Zornoza, A., Serrano, S. and Alonso, J.L. (2017) Environmental Ordination of Filamentous Bacteria in Activated Sludge. In: Abstracts of the 7th congress of European microbiologists FEMS 2017, Valencia, Spain, 9-13 July 2017.
his study investigated the microbial community in a full scale anaerobic baffled reactor and sequencing batch reactor system for oil-produced water treatment in summer and winter. The community structures of fungi and bacteria were analyzed through polymerase chain reaction–denaturing gradient gel electrophoresis and Illumina high-throughput sequencing, respectively. Chemical oxygen demand effluent concentration achieved lower than 50 mg/L level after the system in both summer and winter, however, chemical oxygen demand removal rates after anaerobic baffled reactor treatment system were significant higher in summer than that in winter, which conformed to the microbial community diversity. Saccharomycotina, Fusarium, and Aspergillus were detected in both anaerobic baffled reactor and sequencing batch reactor during summer and winter. The fungal communities in anaerobic baffled reactor and sequencing batch reactor were shaped by seasons and treatment units, while there was no correlation between abundance of fungi and chemical oxygen demand removal rates. Compared to summer, the total amount of the dominant hydrocarbon degrading bacteria decreased by 10.2% in anaerobic baffled reactor, resulting in only around 23% of chemical oxygen demand was removed in winter. Although microbial community significantly varied in the three parallel sulfide reducing bacteria, the performance of these bioreactors had no significant difference between summer and winter.
2008 - Molecular microbial and chemical investigation of the bioremediation o...WALEBUBLÉ
The document describes a study that used laboratory-scale bioreactors to investigate the biodegradation of two-phase olive mill waste (TPOMW) by its indigenous microbiota under different conditions. The effects of nutrient addition and aeration on bioremediation and microbial community changes were evaluated. Analysis found that nutrient addition and aeration led to greater decreases in polyphenolic content and increases in the fungal to bacterial ratio. Molecular identification of bacteria and fungi in the bioreactors identified several genera present, with fungi like Penicillium and Candida dominant.
This document summarizes a study on the uptake of pharmaceuticals by vegetable crops during irrigation with recycled water. Key findings include:
- Crops were found to uptake selected pharmaceuticals (atenolol, ofloxacin, diclofenac) from soil into edible parts at low ng/g levels.
- Uptake depended on soil moisture levels, with higher uptake at field capacity. Ofloxacin showed the highest uptake levels.
- Higher input concentrations of pharmaceuticals in irrigation water led to higher residues in plant tissues.
- Experiments were conducted in Hawaii and Arizona on various crops like basil, lettuce, peppers under different soil moisture and pharmaceutical concentration conditions.
- Further work is needed
This document summarizes a research project with 4 main tasks: [1] generate experimental data from algae bioreactors and calibrate optimization models, [2] maximize the nutritional value of algae for animal feed, [3] optimize pathogen inactivation methods, and [4] quantify and control toxic cyanobacteria. For task 1, the researchers collected seasonal data from outdoor bioreactors to develop productivity models. They identified dominant algae species and established laboratory protocols. For task 2, they isolated algae strains to optimize growth and nutritional composition. Tasks 3 and 4 will use these results to optimize pathogen inactivation and detect/control cyanotoxins.
2017 - Analysis of nitrifying microbial communities by FISH and 16S rRNA ampl...WALEBUBLÉ
Nitrification, the sequential oxidation of ammonia via nitrite to nitrate, is an important process for nitrogen removal from municipal wastewater. This process is catalysed by ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB), two different groups of slow-growing microorganisms whose cooperation is needed to achieve complete nitrification. High efficiency and stability of this process is required for wastewater treatment plants (WWTPs) operational optimization due to
nitrification is often subjected to recurring collapse in many WWTPs. Therefore, a better understanding of the microbial ecology of nitrifying bacteria in WWTPs could
potentially improve the nitrification stability. Novel high-throughput molecular methods, as next generation sequencing (NGS), are nowadays providing detailed knowledge on the microorganisms governing wastewater treatment systems. This
methods in conjunction with the environmental ordination of the relationships between biological variables (nitrifying bacterial community) and physicochemical variables (nitrogen compounds and environmental conditions) provide a powerful
tool to elucidate how selection pressures imposed by operational and environmental conditions affect community diversity and dynamics within activated sludge systems.
2017 - Effect of ozone addition to control Gordonia foaming on the nitrifying...WALEBUBLÉ
The ozonation of activated sludge has been used as a technical measure for bulking control in a high number of full-scale wastewater treatment plants (WWTP), despite a lack of precise
predictions on the level of reduction in filament growth or the lack of knowledge of impact on microbial community from this technique. Ozone is a strong oxidant reacting rapidly with
suspended solids. Various studies have suggested that ozone attacks the bacterial cell surface, alters the permeability of the cell membrane and ultimately results in the leakage of cell
contents. However, the microbes in the sludge form a complex matrix, and ozone may affect bacterial populations at different rates different depending on their locations in the floc or their
capacity for adaptation. Nitrification, a key step of the nitrogen cycle, is the sequential oxidation of ammonia via nitrite to nitrate. This process is catalysed by ammonia-oxidizing bacteria
(AOB) and nitrite-oxidizing bacteria (NOB), whose cooperation is needed to achieve complete nitrification. Although the nitrification process in WWTPs has been investigated in depth, the response of microbial communities are still a focus of considerable interest due to their high sensitivity to inhibitory compounds and environmental factors that results in repeated
breakdowns of nitrification performance. In this study, we focus on two aspects that have not been thoroughly considered in previous studies; the use of ozone for Gordonia foaming
elimination on dynamic population of a nitrifying bacterial community, and the nitrification performance of activated sludge system.
This document summarizes a research study that used an anoxic-aerobic sequencing batch reactor (SBR) system to treat high-strength wastewater containing 1000 mg/L of nitrate and 4000 mg/L of chemical oxygen demand (COD). The SBR was able to simultaneously remove 98% of nitrate, 86% of phosphate, and 72% of COD after 180 days of operation. Pyrosequencing analysis of the microbial communities revealed that Proteobacteria, Alphaproteobacteria, Rhodobacterales, Rhodobacteraceae, and Paracoccous were the dominant taxa present. The surplus electron donors and acceptors in the anoxic phase helped enrich denitrifying phosphate accumulating organisms, while
This document discusses using phosphate oxygen isotope ratios (δ18OP) to better understand phosphorus cycling in agricultural soils. It presents the goals of developing δ18OP as a tracer to identify the bioavailable P fraction in soils and track the long-term fate of externally applied P. It describes sample processing methods, measurement techniques, and initial findings showing transformation of fertilizer P into recalcitrant apatite P pools in agricultural soils. The document concludes that stable isotope labeling and tracking allows a deeper understanding of P sources, transfer, and transformations in natural environments.
Van Houten, 2009, Desulfovibrio Paquesii Sp. Nov., A Hydrogenotrophic Sulfate...roelmeulepas
This document describes the isolation and characterization of a novel sulfate-reducing bacterium, strain SB1T, isolated from a synthesis-gas-fed bioreactor treating zinc- and sulfate-rich wastewater. 16S rRNA gene sequencing showed that strain SB1T was most closely related to Desulfovibrio gigas, with 97.5% sequence similarity. However, DNA-DNA hybridization showed only 56% relatedness between the two strains, below the threshold for species designation. Phenotypic characterization also differentiated strain SB1T from D. gigas. Therefore, a new species, Desulfovibrio paquesii sp. nov., is proposed to accommodate strain SB1T.
2018 - Multiple identification of most important waterborne protozoa in surfa...WALEBUBLÉ
This study evaluated a next-generation sequencing (NGS)-based method to simultaneously identify multiple important waterborne protozoan parasites (WPPs) using 18S rRNA gene amplicon sequencing. The method used a set of primers targeting the V4 region of WPPs such as Cryptosporidium, Giardia, Blastocystis, Entamoeba, Toxoplasma, and free-living amoebae. The primers were tested on a mock community and surface water sample spiked with WPP DNA. The optimized method detected Giardia intestinalis, Acanthamoeba castellanii, Toxoplasma gondii, Entamoeba histoly
2015 - Archaeal populations in full-scale autotrophic nitrogen removal biorea...WALEBUBLÉ
This study analyzed the archaeal populations present in three full-scale autotrophic nitrogen removal bioreactors located in the Netherlands with different technologies: partial nitritation/anammox, CANON, and DEMON. Quantitative PCR and 16S rRNA gene pyrosequencing revealed the presence of archaea in all three bioreactors, comprising over 0.3% of the total prokaryotic communities. Phylogenetic analysis identified archaeal species related to methanogenesis and nitrogen cycling. The results confirm archaea play a role in full-scale anammox processes, though their functions require further study. An inverse correlation was also found between archaeal abundance and bioreactor performance.
This document describes research on nitrogen transformations in aquaponic systems. The objectives were to quantify the impact of physical and chemical variables on nitrogen transformations, evaluate transformations of different nitrogen forms under various conditions, examine ecologically important species, and investigate greenhouse gas emissions. Experiments were conducted with different hydraulic loading rates, pH levels, plant species, and sediment draining frequencies. The results showed that nitrate accumulation was affected by loading rate, pH, and plant uptake rate, while nitrogen loss occurred through denitrification. Microbial analyses provided insight into nitrifying bacteria populations and their role in the nitrogen cycle.
Molecular exploration of biomarkers as early warning system of aquatic pollutionAlexander Decker
1. The document discusses a study analyzing molecular changes in hepatocytes of carp (Cyprinus carpio L.) exposed to various concentrations of linear alkylbenzene sulfonate (LAS), a common pollutant found in aquatic environments.
2. Biomarkers like heat shock protein 70 (HSP70), inducible nitric oxide synthase (iNOS), p38 mitogen-activated protein kinase (p38 MAPK), and cytochrome P450 1A (CYP1A) were observed in hepatocytes using immunohistochemical methods.
3. The results showed a significant increase in the expression of all biomarkers in hepatocytes with increasing LAS concentration and exposure time, indicating that these biomarkers can
4.11 - "Aquatic biomass as a source of renewable energy" - Miroslaw Krzemiene...Pomcert
The document summarizes research on using algal biomass as a source of renewable energy. It describes the history of using algae for biofuel production and discusses cultivation of algae in open and closed systems. A pilot plant was constructed on the Vistula Lagoon to separate algal biomass using a floatation and filtration system. Tests of biogas production from the algal biomass showed the highest efficiency at organic compound loadings of 1.0 to 2.0 kg/m3/day, yielding up to 400 m3 of biogas per ton of dry mass.
This document summarizes research on how microbial activities control phosphorus mobility in agroecosystems under changing redox conditions. Studies examined stream biofilms and soils. Bench tests showed stream biofilms released phosphorus under anaerobic conditions due to polyphosphate-accumulating organisms (PAOs), and took it up aerobically. PAOs comprised 13-38% of biofilm cells. Soil columns will uncouple contributions of dissimilatory iron-reducing bacteria and PAOs to phosphorus trends under variable saturation. Ongoing work analyzes soils across saturation frequencies and metagenomes of microbial communities.
This document reports on the isolation and characterization of a denitrifying Acinetobacter baumannii strain H1 that was isolated from shrimp farming ponds. Strain H1 was able to use nitrite as its sole nitrogen source and showed high nitrite removal rates. It was identified as A. baumannii through biochemical testing and phylogenetic analysis of its 16S rRNA gene. PCR analysis detected key denitrification genes in strain H1. In vitro experiments demonstrated that strain H1 was able to remove nitrite effectively over a wide range of pH, temperatures, and cell densities. Tests also showed that strain H1 was non-pathogenic to mammals and shrimp. This study suggests that A. baumannii strain H
This document summarizes a study on the anaerobic digestion of cow dung for biogas production. Cow dung was digested in a 10L bioreactor at 53°C in batch and semi-continuous modes. In batch operation, the biogas yield was highest at 0.15 L/kg VS added with 47% methane content. Total solids and volatile solids were reduced by 49% and 47% respectively. In semi-continuous operation, biogas production and removals decreased likely due to uncontrolled pH and resulting ammonia inhibition. The study shows cow dung is an effective feedstock for anaerobic digestion and biogas production.
2010 - Assessment of advanced wastewater treatments for nitrogen removal sear...WALEBUBLÉ
This document summarizes a study that assessed advanced wastewater treatment plants for nitrogen removal in Spain over 14 months. The study characterized biological communities by identifying and counting protists (ciliates, flagellates, amoeba) and recording physical-chemical parameters. Surprisingly, ciliates were found to be less abundant and diverse than in conventional treatment plants. The aim was to determine potential biological indicators of treatment efficiency, as indicators from conventional plants could not be directly extrapolated to advanced nutrient removal systems with different biological characteristics.
Characterization of microbial communities in water and biofilms along a large...Adi Levi
The document summarizes a study characterizing microbial communities along a large-scale seawater reverse osmosis (SWRO) desalination facility. The study found:
1) Microbial biofilm communities differed significantly between locations in the treatment process (rapid sand filtration, cartridge filters, reverse osmosis membranes).
2) Biofilms that developed on cartridge filters provided inocula that enhanced downstream biofouling on reverse osmosis membranes.
3) Prior to developing advanced antibiofouling treatments, site-specific characterization of microbial communities in feedwater, pretreatment systems, and on reverse osmosis membranes is required to optimize pretreatment and cleaning procedures and reduce costs.
This document summarizes research on the biological and abiotic transformation of contaminants of emerging concern (CECs) in anaerobic soils. The research has three main objectives: 1) study the biodegradation of the anticonvulsant drug carbamazepine (CBZ) in soils, 2) examine abiotic factors influencing CEC degradation, and 3) investigate mineral-catalyzed degradation of organophosphate flame retardants. For CBZ, results show removal only under aerobic conditions in one soil, and enrichment of phylotypes like Sphingomonadaceae linked to degradation. Screening of CEC reactivity with abiotic soil components found reactions with reduced sulfur, iron, and
This document describes an experiment assessing the use of green crabs (Carcinus maenas) as indicator species in the Multispecies Freshwater Biomonitor (MFB), an online water quality biomonitor. The experiment tested whether: 1) the electrical field of the MFB affects crab behavior; 2) ammonia presence alters crab behavior; 3) the electrical field further impacts behavior in ammonia's presence. Results showed the electrical field did not significantly impact crabs on its own. Ammonia did alter crab behavior in a dose-dependent manner. Five distinct crab behaviors were detected by the MFB. Overall, green crabs appear suitable for use in the MFB as marine biomonitors.
This study analyzed bacterial communities in drinking water biofilms using next-generation sequencing of 16S rRNA genes. Biofilm samples were collected from water meters and pipes in a drinking water distribution system in southern Sweden. Over 600,000 DNA sequences were obtained and classified. The bacterial communities differed between water meters from households with and without complaints about water quality. Water meters from complaining households had fewer Proteobacteria and more Nitrospira and Pedomicrobium. Biofilm communities also differed between water meters and pipes, with pipes containing more Mycobacterium, Nocardia, Desulfovibrio, and Sulfuricurvum. Next-generation sequencing resolved bacterial diversity and differences in communities associated with water quality
2009 - Efficiency of nitrogen removal and protist communities the potential f...WALEBUBLÉ
Article published in the International Workshop on Integrated vision of urban and agro-industrial wastewater treatment, monitoring and reclamation: key role played by the Waste Water Treatment Plant. 2-3 Julio, 2009. ISRIM / LIFE (CEE n. 1973/92 EU Financial Instrument for the Environment) , Terni, Italy.
This document summarizes a study that used a continuous stirred bed reactor (SBR) to treat textile wastewater through bacterial decolourisation. A mixture of three bacterial strains - Sphingomonas paucimobilis, Bacillus sp., and filamentous bacteria - were introduced to the SBR. The study used a mixture experimental design to optimize the formulation of bacterial strains and assess colour and COD removal. The results found that the highest predictable decolourisation and COD removal rates were 86.72% and 75.06% respectively. Regression analysis showed excellent correlation between the bacterial mixtures and responses of decolourisation and COD removal.
This document summarizes a study on the uptake of pharmaceuticals by vegetable crops during irrigation with recycled water. Key findings include:
- Crops were found to uptake selected pharmaceuticals (atenolol, ofloxacin, diclofenac) from soil into edible parts at low ng/g levels.
- Uptake depended on soil moisture levels, with higher uptake at field capacity. Ofloxacin showed the highest uptake levels.
- Higher input concentrations of pharmaceuticals in irrigation water led to higher residues in plant tissues.
- Experiments were conducted in Hawaii and Arizona on various crops like basil, lettuce, peppers under different soil moisture and pharmaceutical concentration conditions.
- Further work is needed
This document summarizes a research project with 4 main tasks: [1] generate experimental data from algae bioreactors and calibrate optimization models, [2] maximize the nutritional value of algae for animal feed, [3] optimize pathogen inactivation methods, and [4] quantify and control toxic cyanobacteria. For task 1, the researchers collected seasonal data from outdoor bioreactors to develop productivity models. They identified dominant algae species and established laboratory protocols. For task 2, they isolated algae strains to optimize growth and nutritional composition. Tasks 3 and 4 will use these results to optimize pathogen inactivation and detect/control cyanotoxins.
2017 - Analysis of nitrifying microbial communities by FISH and 16S rRNA ampl...WALEBUBLÉ
Nitrification, the sequential oxidation of ammonia via nitrite to nitrate, is an important process for nitrogen removal from municipal wastewater. This process is catalysed by ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB), two different groups of slow-growing microorganisms whose cooperation is needed to achieve complete nitrification. High efficiency and stability of this process is required for wastewater treatment plants (WWTPs) operational optimization due to
nitrification is often subjected to recurring collapse in many WWTPs. Therefore, a better understanding of the microbial ecology of nitrifying bacteria in WWTPs could
potentially improve the nitrification stability. Novel high-throughput molecular methods, as next generation sequencing (NGS), are nowadays providing detailed knowledge on the microorganisms governing wastewater treatment systems. This
methods in conjunction with the environmental ordination of the relationships between biological variables (nitrifying bacterial community) and physicochemical variables (nitrogen compounds and environmental conditions) provide a powerful
tool to elucidate how selection pressures imposed by operational and environmental conditions affect community diversity and dynamics within activated sludge systems.
2017 - Effect of ozone addition to control Gordonia foaming on the nitrifying...WALEBUBLÉ
The ozonation of activated sludge has been used as a technical measure for bulking control in a high number of full-scale wastewater treatment plants (WWTP), despite a lack of precise
predictions on the level of reduction in filament growth or the lack of knowledge of impact on microbial community from this technique. Ozone is a strong oxidant reacting rapidly with
suspended solids. Various studies have suggested that ozone attacks the bacterial cell surface, alters the permeability of the cell membrane and ultimately results in the leakage of cell
contents. However, the microbes in the sludge form a complex matrix, and ozone may affect bacterial populations at different rates different depending on their locations in the floc or their
capacity for adaptation. Nitrification, a key step of the nitrogen cycle, is the sequential oxidation of ammonia via nitrite to nitrate. This process is catalysed by ammonia-oxidizing bacteria
(AOB) and nitrite-oxidizing bacteria (NOB), whose cooperation is needed to achieve complete nitrification. Although the nitrification process in WWTPs has been investigated in depth, the response of microbial communities are still a focus of considerable interest due to their high sensitivity to inhibitory compounds and environmental factors that results in repeated
breakdowns of nitrification performance. In this study, we focus on two aspects that have not been thoroughly considered in previous studies; the use of ozone for Gordonia foaming
elimination on dynamic population of a nitrifying bacterial community, and the nitrification performance of activated sludge system.
This document summarizes a research study that used an anoxic-aerobic sequencing batch reactor (SBR) system to treat high-strength wastewater containing 1000 mg/L of nitrate and 4000 mg/L of chemical oxygen demand (COD). The SBR was able to simultaneously remove 98% of nitrate, 86% of phosphate, and 72% of COD after 180 days of operation. Pyrosequencing analysis of the microbial communities revealed that Proteobacteria, Alphaproteobacteria, Rhodobacterales, Rhodobacteraceae, and Paracoccous were the dominant taxa present. The surplus electron donors and acceptors in the anoxic phase helped enrich denitrifying phosphate accumulating organisms, while
This document discusses using phosphate oxygen isotope ratios (δ18OP) to better understand phosphorus cycling in agricultural soils. It presents the goals of developing δ18OP as a tracer to identify the bioavailable P fraction in soils and track the long-term fate of externally applied P. It describes sample processing methods, measurement techniques, and initial findings showing transformation of fertilizer P into recalcitrant apatite P pools in agricultural soils. The document concludes that stable isotope labeling and tracking allows a deeper understanding of P sources, transfer, and transformations in natural environments.
Van Houten, 2009, Desulfovibrio Paquesii Sp. Nov., A Hydrogenotrophic Sulfate...roelmeulepas
This document describes the isolation and characterization of a novel sulfate-reducing bacterium, strain SB1T, isolated from a synthesis-gas-fed bioreactor treating zinc- and sulfate-rich wastewater. 16S rRNA gene sequencing showed that strain SB1T was most closely related to Desulfovibrio gigas, with 97.5% sequence similarity. However, DNA-DNA hybridization showed only 56% relatedness between the two strains, below the threshold for species designation. Phenotypic characterization also differentiated strain SB1T from D. gigas. Therefore, a new species, Desulfovibrio paquesii sp. nov., is proposed to accommodate strain SB1T.
2018 - Multiple identification of most important waterborne protozoa in surfa...WALEBUBLÉ
This study evaluated a next-generation sequencing (NGS)-based method to simultaneously identify multiple important waterborne protozoan parasites (WPPs) using 18S rRNA gene amplicon sequencing. The method used a set of primers targeting the V4 region of WPPs such as Cryptosporidium, Giardia, Blastocystis, Entamoeba, Toxoplasma, and free-living amoebae. The primers were tested on a mock community and surface water sample spiked with WPP DNA. The optimized method detected Giardia intestinalis, Acanthamoeba castellanii, Toxoplasma gondii, Entamoeba histoly
2015 - Archaeal populations in full-scale autotrophic nitrogen removal biorea...WALEBUBLÉ
This study analyzed the archaeal populations present in three full-scale autotrophic nitrogen removal bioreactors located in the Netherlands with different technologies: partial nitritation/anammox, CANON, and DEMON. Quantitative PCR and 16S rRNA gene pyrosequencing revealed the presence of archaea in all three bioreactors, comprising over 0.3% of the total prokaryotic communities. Phylogenetic analysis identified archaeal species related to methanogenesis and nitrogen cycling. The results confirm archaea play a role in full-scale anammox processes, though their functions require further study. An inverse correlation was also found between archaeal abundance and bioreactor performance.
This document describes research on nitrogen transformations in aquaponic systems. The objectives were to quantify the impact of physical and chemical variables on nitrogen transformations, evaluate transformations of different nitrogen forms under various conditions, examine ecologically important species, and investigate greenhouse gas emissions. Experiments were conducted with different hydraulic loading rates, pH levels, plant species, and sediment draining frequencies. The results showed that nitrate accumulation was affected by loading rate, pH, and plant uptake rate, while nitrogen loss occurred through denitrification. Microbial analyses provided insight into nitrifying bacteria populations and their role in the nitrogen cycle.
Molecular exploration of biomarkers as early warning system of aquatic pollutionAlexander Decker
1. The document discusses a study analyzing molecular changes in hepatocytes of carp (Cyprinus carpio L.) exposed to various concentrations of linear alkylbenzene sulfonate (LAS), a common pollutant found in aquatic environments.
2. Biomarkers like heat shock protein 70 (HSP70), inducible nitric oxide synthase (iNOS), p38 mitogen-activated protein kinase (p38 MAPK), and cytochrome P450 1A (CYP1A) were observed in hepatocytes using immunohistochemical methods.
3. The results showed a significant increase in the expression of all biomarkers in hepatocytes with increasing LAS concentration and exposure time, indicating that these biomarkers can
4.11 - "Aquatic biomass as a source of renewable energy" - Miroslaw Krzemiene...Pomcert
The document summarizes research on using algal biomass as a source of renewable energy. It describes the history of using algae for biofuel production and discusses cultivation of algae in open and closed systems. A pilot plant was constructed on the Vistula Lagoon to separate algal biomass using a floatation and filtration system. Tests of biogas production from the algal biomass showed the highest efficiency at organic compound loadings of 1.0 to 2.0 kg/m3/day, yielding up to 400 m3 of biogas per ton of dry mass.
This document summarizes research on how microbial activities control phosphorus mobility in agroecosystems under changing redox conditions. Studies examined stream biofilms and soils. Bench tests showed stream biofilms released phosphorus under anaerobic conditions due to polyphosphate-accumulating organisms (PAOs), and took it up aerobically. PAOs comprised 13-38% of biofilm cells. Soil columns will uncouple contributions of dissimilatory iron-reducing bacteria and PAOs to phosphorus trends under variable saturation. Ongoing work analyzes soils across saturation frequencies and metagenomes of microbial communities.
This document reports on the isolation and characterization of a denitrifying Acinetobacter baumannii strain H1 that was isolated from shrimp farming ponds. Strain H1 was able to use nitrite as its sole nitrogen source and showed high nitrite removal rates. It was identified as A. baumannii through biochemical testing and phylogenetic analysis of its 16S rRNA gene. PCR analysis detected key denitrification genes in strain H1. In vitro experiments demonstrated that strain H1 was able to remove nitrite effectively over a wide range of pH, temperatures, and cell densities. Tests also showed that strain H1 was non-pathogenic to mammals and shrimp. This study suggests that A. baumannii strain H
This document summarizes a study on the anaerobic digestion of cow dung for biogas production. Cow dung was digested in a 10L bioreactor at 53°C in batch and semi-continuous modes. In batch operation, the biogas yield was highest at 0.15 L/kg VS added with 47% methane content. Total solids and volatile solids were reduced by 49% and 47% respectively. In semi-continuous operation, biogas production and removals decreased likely due to uncontrolled pH and resulting ammonia inhibition. The study shows cow dung is an effective feedstock for anaerobic digestion and biogas production.
2010 - Assessment of advanced wastewater treatments for nitrogen removal sear...WALEBUBLÉ
This document summarizes a study that assessed advanced wastewater treatment plants for nitrogen removal in Spain over 14 months. The study characterized biological communities by identifying and counting protists (ciliates, flagellates, amoeba) and recording physical-chemical parameters. Surprisingly, ciliates were found to be less abundant and diverse than in conventional treatment plants. The aim was to determine potential biological indicators of treatment efficiency, as indicators from conventional plants could not be directly extrapolated to advanced nutrient removal systems with different biological characteristics.
Characterization of microbial communities in water and biofilms along a large...Adi Levi
The document summarizes a study characterizing microbial communities along a large-scale seawater reverse osmosis (SWRO) desalination facility. The study found:
1) Microbial biofilm communities differed significantly between locations in the treatment process (rapid sand filtration, cartridge filters, reverse osmosis membranes).
2) Biofilms that developed on cartridge filters provided inocula that enhanced downstream biofouling on reverse osmosis membranes.
3) Prior to developing advanced antibiofouling treatments, site-specific characterization of microbial communities in feedwater, pretreatment systems, and on reverse osmosis membranes is required to optimize pretreatment and cleaning procedures and reduce costs.
This document summarizes research on the biological and abiotic transformation of contaminants of emerging concern (CECs) in anaerobic soils. The research has three main objectives: 1) study the biodegradation of the anticonvulsant drug carbamazepine (CBZ) in soils, 2) examine abiotic factors influencing CEC degradation, and 3) investigate mineral-catalyzed degradation of organophosphate flame retardants. For CBZ, results show removal only under aerobic conditions in one soil, and enrichment of phylotypes like Sphingomonadaceae linked to degradation. Screening of CEC reactivity with abiotic soil components found reactions with reduced sulfur, iron, and
This document describes an experiment assessing the use of green crabs (Carcinus maenas) as indicator species in the Multispecies Freshwater Biomonitor (MFB), an online water quality biomonitor. The experiment tested whether: 1) the electrical field of the MFB affects crab behavior; 2) ammonia presence alters crab behavior; 3) the electrical field further impacts behavior in ammonia's presence. Results showed the electrical field did not significantly impact crabs on its own. Ammonia did alter crab behavior in a dose-dependent manner. Five distinct crab behaviors were detected by the MFB. Overall, green crabs appear suitable for use in the MFB as marine biomonitors.
This study analyzed bacterial communities in drinking water biofilms using next-generation sequencing of 16S rRNA genes. Biofilm samples were collected from water meters and pipes in a drinking water distribution system in southern Sweden. Over 600,000 DNA sequences were obtained and classified. The bacterial communities differed between water meters from households with and without complaints about water quality. Water meters from complaining households had fewer Proteobacteria and more Nitrospira and Pedomicrobium. Biofilm communities also differed between water meters and pipes, with pipes containing more Mycobacterium, Nocardia, Desulfovibrio, and Sulfuricurvum. Next-generation sequencing resolved bacterial diversity and differences in communities associated with water quality
2009 - Efficiency of nitrogen removal and protist communities the potential f...WALEBUBLÉ
Article published in the International Workshop on Integrated vision of urban and agro-industrial wastewater treatment, monitoring and reclamation: key role played by the Waste Water Treatment Plant. 2-3 Julio, 2009. ISRIM / LIFE (CEE n. 1973/92 EU Financial Instrument for the Environment) , Terni, Italy.
This document summarizes a study that used a continuous stirred bed reactor (SBR) to treat textile wastewater through bacterial decolourisation. A mixture of three bacterial strains - Sphingomonas paucimobilis, Bacillus sp., and filamentous bacteria - were introduced to the SBR. The study used a mixture experimental design to optimize the formulation of bacterial strains and assess colour and COD removal. The results found that the highest predictable decolourisation and COD removal rates were 86.72% and 75.06% respectively. Regression analysis showed excellent correlation between the bacterial mixtures and responses of decolourisation and COD removal.
This document summarizes a study on enhancing biogas production from abattoir cow liquor waste and various agro-industrial wastes through anaerobic digestion. Cow liquor waste was combined with brewery spent grain, cassava waste water, and carbonated soft drink sludge at different ratios. Adding cow liquor waste significantly increased biogas production from brewery spent grain but had negative effects on carbonated soft drink sludge. Cassava waste water, which previously did not produce biogas, was able to produce biogas when inoculated with cow liquor waste. The study suggests cow liquor waste can improve biogas yields from some agro-industrial wastes through anaerobic digestion.
This document presents a study on quantifying the neurotoxic cyanobacterial toxin β-N-methylamino-L-alanine (BMAA) in irrigation water sources, fish, and crop plants in Sohag governorate, Egypt. Water, phytoplankton, fish, and plant samples were collected from various sites and analyzed for free and protein-bound BMAA using extraction and HPLC-MS/MS methods. The objectives were to detect BMAA in these matrices, identify cyanobacteria producing BMAA, and determine BMAA concentrations in fish and plant tissues to evaluate potential health risks to humans. The results of this study could provide evidence of BMAA accumulation in Egyptian freshwaters, fish, and agricultural crops
This document is a thesis project examining innovative biological phosphate and anaerobic digestion technology for waste treatment, energy generation, and phosphorus recovery. It includes an abstract, introduction covering topics like sewage, current bioremediation methods, anaerobic digestion processes, low temperature anaerobic digestion, bioreactor configurations, and the global phosphorus crisis. It also outlines the materials and methods, expected results sections, and planned discussion. The introduction provides background on anaerobic digestion and examines its application to low-temperature wastewater treatment.
This document describes a project to design an energy-producing waste treatment system using anaerobic co-digestion of organic wastes from the University of Arkansas Swine and Poultry Units coupled with algae cultivation. A prototype was constructed and tested to generate data for designing a full-scale system. The full-scale system was designed to treat all biological wastes from the units while producing net energy and retaining nutrients that could be used as fertilizer.
Compost tea is being used increasingly in agricultural production to
control plant diseases. However, there has been limited investigation relating
disease control efficacy to various compost tea production methods,
particularly compost tea produced with active aeration and additives to
increase microbial population densities in compost tea. Aerated compost
tea (ACT) and nonaerated compost tea (NCT), produced with or without
additives, was investigated for the suppression of damping-off of cucumber
caused by Pythium ultimum. Compost tea was used to drench soilless
container medium inoculated with P. ultimum; effect on damping-off
ranged from not suppressive to consistently suppressive depending on the
method used to produce the tea. The most consistent formulation for
damping-off suppression was ACT produced with kelp and humic acid
additives. Producing ACT with a molasses-based additive inconsistently
suppressed damping-off; evidence suggests that residual nutrients can interfere
with disease suppression. Heating or diluting compost tea negated
suppression. Across all compost tea samples, there was no significant
relationship of bacterial populations, measured as active cells, total cells,
or CFU, to disease suppression. However, for all ACT produced without
the molasses-based additive, there was a threshold of bacterial population
density (6 log10 active cells per ml, 7.48 log10 total cells per ml, or 7 log10
CFU per ml) above which compost teas were suppressive.
Do pitcher plants control the assembly of pitcher microbiomes?Leonora Bittleston
The document summarizes a study investigating the factors influencing the assembly of microbial communities within the pitchers of three Nepenthes plant species in Singapore. The study finds that bacterial and eukaryotic community composition in the pitchers is most influenced by the host plant species and pH level. While collection site and pitcher volume also have significant effects, bacterial and eukaryotic diversity are strongly correlated, suggesting similar responses to environmental conditions rather than interactions between organisms shaping the communities. Deterministic factors like biochemical characteristics have a stronger influence on community assembly than stochastic factors like chance colonization.
Cashew nut processing industry waste water treatmentVishnu Raj
This document discusses the treatment of waste water from the cashew nut processing industry in India. It provides background on the industry and production levels in India. It then describes the sources and characteristics of the wastewater, which contains toxic cashew nut shell liquid (CNSL) that is difficult to treat using conventional biological methods. The document reviews various treatment alternatives that have been studied including aerobic reactors with fungal inoculums, bioaugmentation with specific microorganisms, and enzymatic bioremediation approaches. It concludes that biological remediation technologies show promise for treating this type of wastewater but further optimization is needed given the recalcitrant nature of some CNSL components.
International Journal of Engineering and Science Invention (IJESI) inventionjournals
International Journal of Engineering and Science Invention (IJESI) is an international journal intended for professionals and researchers in all fields of computer science and electronics. IJESI publishes research articles and reviews within the whole field Engineering Science and Technology, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Investigation of the effect of initial biomass on nitrate and phosphate remov...Alexander Decker
This study investigated the effect of initial biomass concentration on the ability of four bacterial species
(Klebsiella sp., Pseudomonas sp., Lysinibacillus sp., and Staphylococcus sp.) to remove nitrate and phosphate from synthetic wastewater. The bacteria were inoculated at different concentrations and their ability to remove nutrients was measured over 96 hours. All isolates significantly removed nitrate except Lysinibacillus sp., removing between 68-91% nitrate. Phosphate removal was slight. The study revealed the nutrient removal abilities of the isolates at different initial biomass concentrations.
High Rate of Water Biodenitrification Using Anthracite as Hyphomicrobium Deni...theijes
Pure culture of Hyphomicrobium denitrificans DSM 1869 was immobilized on anthracite and utilized for biological denitrification in 50-ml flasks employing methanol and acetic acid as carbon source. The results demonstrate that acetic acid was a suitable carbon source for H. denitrificans to remove high nitrate concentrations. The maximum denitrification rate was 233.1 mg NO3-N/g MLSS.h and the highest NO3-N removal efficiency was obtained when using C/N ratio of 4.0 and acetic acid as the carbon source. C/N ratio can significantly affect denitrification in different operational conditions. The low C/N ratios did not allow the denitrification process to be completed in case of high NO3-Nconcentrations. High C/N ratio increased the rate of nitrate conversion when using acetic acid as a carbon source; but added a pollutant to denitrified water when using methanol as a carbon source. The results demonstrated that H. denitrificans was a suitable bacterium for denitrifying high NO3-N concentrations.
Slaughter waste effluents and river catchment watershed contamination in Caga...Angelo Mark Walag
Slaughterhouse waste products are commonly known globally to pollute nearby communities and receiving bodies of water. The main aim of this study was to analyze the effluents disposed by Cagayan de Oro City Slaughterhouse to river catchment watershed. Standard methods were utilized in sampling and analyzing water quality parameters to determine the levels of nitrates, BOD, COD, total coliform, and lead. It was found out that the majority of wastes produced are internal organs, blood and urine mixtures, and manures. The study also revealed that all parameters tested crossed the permissible limits set by the government for effluent and inland water except for BOD and nitrates, in the river watershed. It was also determined that during wet seasons, major contaminants like lead and nitrates were diluted resulting to lower levels when compared to national standards. The result of this study also revealed the need for further remediation of the river water quality and intervention strategies to sustainably manage and prevent disposal of untreated effluents.
ANABAENA AZOLLAE CULTURE WATER, A SOURCE FOR BIOREMEDIATIONanupfpic
ABSTRACT:
The Azolla-Anabaena symbiosis is outstanding due to its high productivity combined with its ability to fix nitrogen
at high rates. Anabaena azollae a blue green alga, isolated from a pteridophyte, Azolla pinnata by growing in stress
conditions. Anabaena culture was grown in Benecks media and further transferred in sterile soil and tap water
medium (w/v). Water from the cultures of Anabaena contains nitrogen and can promote the growth of test crop.
Taken this into consideration, the bioassays were performed on growth of wheat by evaluating different growth
parameters, like germination percentage, seedling height, weight and chlorophyll content. The effect of Anabaena
culture on soil texture was observed by analyzing the physico-chemical properties of treated soil against the
control. In the experiment of presoaking treatment of wheat seeds, 12 hrs presoaking was found most promoting
which showed 100% germination. Wheat seedlings growth enhancement was observed in treated wheat seedling
with Anabaena culture water and a significant improvement in nitrogen, potassium, phosphorus content was
observed with increased pH and reduced EC. The Anabaena culture water was analyzed in comparison with
normal tap water as control, to evaluate the difference in the nutrient quality available for treated plants.
Key words: - Anabaena azollae, Azolla pinnata, Bioremediation, Blue green algaABSTRACT:
The Azolla-Anabaena symbiosis is outstanding due to its high productivity combined with its ability to fix nitrogen
at high rates. Anabaena azollae a blue green alga, isolated from a pteridophyte, Azolla pinnata by growing in stress
conditions. Anabaena culture was grown in Benecks media and further transferred in sterile soil and tap water
medium (w/v). Water from the cultures of Anabaena contains nitrogen and can promote the growth of test crop.
Taken this into consideration, the bioassays were performed on growth of wheat by evaluating different growth
parameters, like germination percentage, seedling height, weight and chlorophyll content. The effect of Anabaena
culture on soil texture was observed by analyzing the physico-chemical properties of treated soil against the
control. In the experiment of presoaking treatment of wheat seeds, 12 hrs presoaking was found most promoting
which showed 100% germination. Wheat seedlings growth enhancement was observed in treated wheat seedling
with Anabaena culture water and a significant improvement in nitrogen, potassium, phosphorus content was
observed with increased pH and reduced EC. The Anabaena culture water was analyzed in comparison with
normal tap water as control, to evaluate the difference in the nutrient quality available for treated plants.
Key words: Anabaena azollae, Azolla pinnata, Bioremediation, Blue green alga
1) The document describes a study on the anaerobic co-digestion of water hyacinth and poultry litter.
2) Batch experiments were conducted in 300ml digesters at mesophilic temperatures to determine biogas production, total solids, volatile solids, and pH over retention periods of 0-56 days.
3) Kinetic modeling using an integral method revealed that the co-digestion process followed first-order reaction kinetics with a rate constant of 0.026 day-1.
IRJET-Biogas Generation from Combination of Food Waste and Water HyacinthIRJET Journal
1. The document discusses biogas generation from anaerobic digestion of various ratios of food waste and water hyacinth using cow dung as a seed material. Ratios studied included 80:20, 70:30, 60:40, and 0:100 of food waste to water hyacinth.
2. For the 80:20 ratio, biogas generation was 450 mL and reductions in parameters like BOD, COD, and TS were 58%, 68%, and 19% respectively. Nutrient levels like phosphates and nitrates increased.
3. For the 70:30 ratio, biogas generation was highest at 480 mL. Reductions in BOD, COD, and TS were
Bacteriological Investigation of Well Water Samples from Selected Market Loca...inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Bacterial indicators and antibiotic resistance of Escherichia coli in groundw...Agriculture Journal IJOEAR
— The aim of this research was to investigate the groundwater quality in El Barreal basin (Córdoba, Argentina), through bacteriological analysis and antibiotic resistance of fecal bacteria indicators and their relationship with geochemical and land uses characteristics. Groundwater samples were collected in 36 wells and the following parameters were determined: major chemical components, heterotrophic plate counts, total and fecal coliforms, Escherichia coli and Pseudomonas aeruginosa. Antibiotic resistance of E. coli was analyzed using standard methods. The chemical and bacteriological analysis showed that more than 80 % of samples were unfit for human consumption. Bacteriological contamination was significant in 36 % of samples. The multivariate analysis between bacterial and geochemical variables explained local contamination conditions, evidenced by the arrival of bacteria and some typical indicators (NO 3-, Cl-y HCO 3-) to groundwater. No significant correlation between NO 3-and bacterial counts was observed, which let us to interpret that part of NO 3-contents can be supplied by inorganic fertilizers. Total and fecal coliforms were linked to a local increase of Cl-and HCO 3-and a decrease of pH and dissolved oxygen, indicating the arrival and degradation of organic matter into groundwater. The resistance pattern of total E. coli isolates (n=12) showed that the highest percentages were observed for antibiotics of animal use (ampicillin, tetracycline and cephalothin). The analysis of results revealed the impact of land uses demonstrating that livestock activities are the main punctual contaminant sources in this sedimentary aquifer.
This document describes a sustainable biorefinery concept that converts animal wastes into value-added products through three main steps:
1) Anaerobic digestion is used to treat animal wastes and produce biogas for energy generation. This results in liquid and solid digestates.
2) The liquid digestate is treated with electrocoagulation to reclaim water.
3) The solid digestate undergoes enzymatic hydrolysis and fungal fermentation using the electrocoagulation water, producing fungal biomass containing the high-value chemical chitin.
The biorefinery concept fully utilizes animal wastes to simultaneously treat wastes and produce a valuable product, while being energy
Investigating the bacterial inactivation potential of purified okra (Hibiscus...AZOJETE UNIMAID
The ability of purified okra protein (POP) as coagulant and as disinfectant material in comparison with aluminium sulphate (AS) in water treatment was assessed. A laboratory jar test experiments and Colilert-18/Quanti-Tray method of bacterial analysis were conducted using POP as coagulant in treating river water. The results show an excellent dual performance function of POP against the conventional coagulant, AS in drinking water treatment. It was observed that a marked inactivation of approximately 100% of faecal and E-coli count in raw water was achieved with POP and zero regrowth of bacteria after 72-hour post treatment. However, there was regrowth in total coliform count as a result of the presence of other microbes other than E-coli and faecal coliform in the system. In all cases AS showed a reduced performance against the two indicator organisms achieving only 93% with remarkable regrowth of E-coli and faecal coliform after prolonged storage time in the clarified water. Turbidity removal was also noted to be approximately similar, 92% across all coagulants tested. Therefore, the use of POP in water treatment could improve access to clean water in developing countries and could help in reducing the import of water treatment chemicals.
Cover Story - China's Investment Leader - Dr. Alyce SUmsthrill
In World Expo 2010 Shanghai – the most visited Expo in the World History
https://www.britannica.com/event/Expo-Shanghai-2010
China’s official organizer of the Expo, CCPIT (China Council for the Promotion of International Trade https://en.ccpit.org/) has chosen Dr. Alyce Su as the Cover Person with Cover Story, in the Expo’s official magazine distributed throughout the Expo, showcasing China’s New Generation of Leaders to the World.
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Dive into this presentation and learn about the ways in which you can buy an engagement ring. This guide will help you choose the perfect engagement rings for women.
The Steadfast and Reliable Bull: Taurus Zodiac Signmy Pandit
Explore the steadfast and reliable nature of the Taurus Zodiac Sign. Discover the personality traits, key dates, and horoscope insights that define the determined and practical Taurus, and learn how their grounded nature makes them the anchor of the zodiac.
Brian Fitzsimmons on the Business Strategy and Content Flywheel of Barstool S...Neil Horowitz
On episode 272 of the Digital and Social Media Sports Podcast, Neil chatted with Brian Fitzsimmons, Director of Licensing and Business Development for Barstool Sports.
What follows is a collection of snippets from the podcast. To hear the full interview and more, check out the podcast on all podcast platforms and at www.dsmsports.net
Digital Marketing with a Focus on Sustainabilitysssourabhsharma
Digital Marketing best practices including influencer marketing, content creators, and omnichannel marketing for Sustainable Brands at the Sustainable Cosmetics Summit 2024 in New York
Discover timeless style with the 2022 Vintage Roman Numerals Men's Ring. Crafted from premium stainless steel, this 6mm wide ring embodies elegance and durability. Perfect as a gift, it seamlessly blends classic Roman numeral detailing with modern sophistication, making it an ideal accessory for any occasion.
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This presentation is a curated compilation of PowerPoint diagrams and templates designed to illustrate 20 different digital transformation frameworks and models. These frameworks are based on recent industry trends and best practices, ensuring that the content remains relevant and up-to-date.
Key highlights include Microsoft's Digital Transformation Framework, which focuses on driving innovation and efficiency, and McKinsey's Ten Guiding Principles, which provide strategic insights for successful digital transformation. Additionally, Forrester's framework emphasizes enhancing customer experiences and modernizing IT infrastructure, while IDC's MaturityScape helps assess and develop organizational digital maturity. MIT's framework explores cutting-edge strategies for achieving digital success.
These materials are perfect for enhancing your business or classroom presentations, offering visual aids to supplement your insights. Please note that while comprehensive, these slides are intended as supplementary resources and may not be complete for standalone instructional purposes.
Frameworks/Models included:
Microsoft’s Digital Transformation Framework
McKinsey’s Ten Guiding Principles of Digital Transformation
Forrester’s Digital Transformation Framework
IDC’s Digital Transformation MaturityScape
MIT’s Digital Transformation Framework
Gartner’s Digital Transformation Framework
Accenture’s Digital Strategy & Enterprise Frameworks
Deloitte’s Digital Industrial Transformation Framework
Capgemini’s Digital Transformation Framework
PwC’s Digital Transformation Framework
Cisco’s Digital Transformation Framework
Cognizant’s Digital Transformation Framework
DXC Technology’s Digital Transformation Framework
The BCG Strategy Palette
McKinsey’s Digital Transformation Framework
Digital Transformation Compass
Four Levels of Digital Maturity
Design Thinking Framework
Business Model Canvas
Customer Journey Map
NIMA2024 | De toegevoegde waarde van DEI en ESG in campagnes | Nathalie Lam |...BBPMedia1
Nathalie zal delen hoe DEI en ESG een fundamentele rol kunnen spelen in je merkstrategie en je de juiste aansluiting kan creëren met je doelgroep. Door middel van voorbeelden en simpele handvatten toont ze hoe dit in jouw organisatie toegepast kan worden.
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Storytelling is an incredibly valuable tool to share data and information. To get the most impact from stories there are a number of key ingredients. These are based on science and human nature. Using these elements in a story you can deliver information impactfully, ensure action and drive change.
1. HRTand nutrients a¡ect bacterial communities grown on
recirculation aquaculture system e¥uents
Oliver Schneider1, Mariana Chabrillon-Popelka2, Hauke Smidt2, Olga Haenen3, Vasiliki Sereti1,
Ep H. Eding1 & Johan A. J. Verreth1
1
Aquaculture and Fisheries Group, Wageningen University, Wageningen, The Netherlands; 2Laboratory of Microbiology, Wageningen University,
Wageningen, The Netherlands; and 3CIDC-Lelystad, NRL for Fish and Shellfish Diseases, Wageningen University, Wageningen, The Netherlands
Correspondence: Oliver Schneider, Abstract
Aquaculture and Fisheries Group,
Wageningen University, PO Box 338, 6700 AH
In a recirculation aquaculture system the drumfilter effluent can be used as
Wageningen, substrate for heterotrophic bacterial production, which can be recycled as feed.
The Netherlands. Tel.: 100 31 317 485147; Because the bacteria might contain pathogens, which could reduce its suitability as
fax: 100 31 317 483937; feed, it is important to characterize these communities. Bacteria were produced in
e-mail: oliver.schneider@wur.nl growth reactors under different conditions: 7 h hydraulic retention time (HRT) vs.
2 h, sodium acetate vs. molasses, and ammonia vs. nitrate. The community of the
Received 26 January 2006; revised 26 drumfilter effluent was different from those found in the reactors. However, all
November 2006; accepted 2 December 2006.
major community components were present in the effluent and reactor broths.
First published online 16 March 2007.
HRT influenced the bacteria community, resulting in a DGGE profile dominated
DOI:10.1111/j.1574-6941.2007.00282.x
by a band corresponding to an Acinetobacter sp.-related population at 2 h HRT
compared to 7 h HRT, where bands indicative of a-proteobacterial populations
Editor: Michael Wagner most closely related to Rhizobium and Shinella spp. were most abundant. Molasses
influenced the bacterial community. It was dominated by an Aquaspirillum
Keywords serpens-related population. Providing total ammonia nitrogen (TAN) in addition
bacteria; community; aquaculture; waste to nitrate led to the occurrence of bacteria close to Sphaerotilus spp., Flavobacter-
conversion; 16S rRNA gene. ium mizutaii and Jonesia spp. It was concluded from these results that a 6–7 h HRT
is recommended, and that the type of substrate is less important, and results in
communities with a comparably low pathogenic risk.
already applied in integrated and activated ponds. In such
Introduction ponds, waste conversion does not only improve water
In recirculation aquaculture systems (RAS), feed is con- quality but also feed conversion ratios, because the pro-
verted into fish and faecal and nonfaecal loss. These two duced bacteria biomass may be consumed by fish or shrimp
waste sources are composed mainly of solid waste, and (Avnimelech et al., 1989; Edwards, 1993; Burford et al., 2003;
dissolved waste: ammonia and phosphate. The waste is Hari et al., 2004).
treated by mechanical filtration to remove the solids from To produce bacterial biomass utilizing the effluent stream
the system water and by biofiltration to nitrify ammonia to of the drum filter, a bacterial reactor has to be integrated
less hazardous nitrate. The effluent from the mechanical into the system (Fig. 1). The nutrient ratios in the slurry
filter is the major discharge of such systems. It comprises coming from the filter are normally not ideal for bacteria
solid (faecal loss) and dissolved waste (nonfaecal loss). This production. Optimal C : N ratios for heterotrophic bacteria
slurry contains dissolved waste, because the solid particles production are about 12–15 g : 1 g (Lechevallier et al., 1991;
are backwashed from the filter screen using system water, Henze et al., 1996; Avnimelech, 1999). Fish receiving high
which contains dissolved waste. The effluent of the RAS is protein diets produce carbon deficient waste. This is due to
either directly discharged to the environment, or digested in the amount of nitrogen, which accumulates in the RAS
lagoons or septic tanks, or thickened and/or applied as system water. The C : N resulting ratio in the effluent is
fertilizer for land based agriculture (Chen et al., 1997; 2–3 g : 1 g (Table 1).
Losordo et al., 2003). A possible alternative approach is to Therefore, the slurry requires organic carbon supplemen-
convert the waste into heterotrophic bacterial biomass. This tation. Sources and levels of carbon supplementation, sludge
biomass can be reutilized as aquatic feed. Such processes are composition [total ammonia nitrogen (TAN) or nitrate]
FEMS Microbiol Ecol 60 (2007) 207–219
c2007 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
2. 208 O. Schneider et al.
Screen filter (60 µm)
Flow
equalizer
Trickling filter
Org. C
Source
pH control
Pure oxygen
Biofilter sump
Bacteria reactor
Pump sump
Bacteria collection
Fig. 1. Simplified scheme of a conventional recirculation aquaculture system for African catfish extended by the bacteria growth reactor and the flow
equalizer.
Table 1. Waste composition measured in the influent of the bioreactors and the influence of different carbon and nitrogen sources
(mg LÀ1) were evaluated. The second objective was to assess if the
Waste concentration produced bacteria biomass contains potential pathogens,
TAN 1.3 Æ 0.8 (0.3–4.8) which could reduce its suitability as feed. To address these
NO2-N 3.3 Æ 1.3 (0.7–12.4) objectives, we used complementary cultivation-dependent
NO3-N 182 Æ 58 (76–419) and 16S rRNA gene-targeted biomolecular approaches,
Kj-N 59 Æ 43 (13–260) aiming at the identification of bacterial populations present
TOC 422 Æ 159 (64–884) under the various process conditions,
Ortho-P 15.1 Æ 7.7 (6.2–40.1)
Ash 1776 Æ 717 (857–4957)
TS 3530 Æ 1033 (1936–7300) Materials and methods
TSS 1472 Æ 1041 (200–5770)
VSS 707 Æ 460 (40–2226) System setup
Conductivity 2000 –3000 mS
Two bacteria growth reactors were connected in parallel to a
Concentrations as averages Æ SD (minimum and maximum).
flow equalizer which received the effluent of a screen filter
TAN, total ammonia nitrogen; NO2-N, nitrite-N; NO3-N, nitrate-N; Kj-N,
(60-mm mesh size, Fig. 1). The screen filter was part of a
Kjeldahl nitrogen corrected for TAN concentrations; TOC, total organic
carbon; ortho-P, ortho-phosphate phosphorus; TS, total solids; TSS, total
recirculation aquaculture system (RAS), which was com-
suspended solids; VSS, volatile suspended solids. posed of four culture tanks, a biofilter and two sumps. In the
equalizer the slurry was aerated and agitated. The equalizer
was integrated into the system to allow for constant waste
flows towards the bacteria reactor, because the screenfilter
and sludge and hydraulic retention time (SRT, HRT) are all backwashes in pulses, depending on its automated flushing
factors influencing the bacteria community forming the cycle. The hydraulic retention time (HRT) of the drumfilter
produced biomass. Furthermore, the community composi- effluent in the equalizer was 4 h and the drumfilter backwash
tion depends also on the natural autochthonous microbiota volume about 120–140 L kgÀ1 feed.
from the sludge and system water. If the produced biomass is
reused as aquatic feed, it is important to evaluate the
Fish husbandry
biomass for potential bacteria pathogens.
The first study objective was to characterize the bacterial Fish were obtained from a commercial African catfish
community in the system water, in the slurry coming from hatchery (Fleuren and Nooijen, The Netherlands). Fish were
the flow equalizer, and of the produced bacterial biomass in stocked initially in four different cohorts of 140 fish each
the reactor. The effect of different hydraulic retention times (70, 170, 320 and 560 g individual average weight) into the
c 2007 Federation of European Microbiological Societies FEMS Microbiol Ecol 60 (2007) 207–219
Published by Blackwell Publishing Ltd. All rights reserved
3. Bacteria communities produced on RAS effluents 209
four tanks. Every 28 days the oldest cohort was harvested. conditions in the reactor (4 2 mg LÀ1). Oxygen was mon-
The emptied tank was restocked with 140 fish of about itored online using pH/Oxi 304i meters (WTW, Germany)
70 g. The final fish weight ranged between 823 and connected to a PC. This PC controlled the oxygenation of
1038 g. Therefore a complete production cycle from 70 to the broth. The oxygen control program reacted on a set-
about 1000 g lasted 112 days. Fish were fed with commercial point concentration of 3 mg LÀ1 oxygen inside the broth. pH
diet (Biomeerval, Skretting, France), containing 7% levels were maintained between 7.0 and 7.2 by addition of
moisture, 49% crude protein, 11% crude fat, 22% carbohy- acid or base (HCl, NaOH, 0.5–1N) stirred by a pH controller
drates, of which 2% crude fiber, 11% crude ash and (Liquisys M, Endress-Hauser, Germany). The reactor tem-
1.7% phosphorous (based on manufacturer information). perature was 28 1C, fixed by a water bath. The reactor was
The realized feeding level was between 16 and 19 g kgÀ1 continuously agitated by a rotor (RZR 2102, Heidolph,
metabolic body weight (W0.8) day-1. Diurnal waste fluctua- Germany) and the agitation speed was fixed to 350 r.p.m.
tions were minimized by applying a 24 h feeding regime. The
monthly harvesting/restocking scheme minimized changes Experimental designs and sampling
in both biomass within the system and in feed load. This
In this study, the bacterial communities corresponding to
stocking and feeding strategy assured minimal fluctuations
the content of bioreactors which operated under four
of waste production during a production cycle.
different conditions were analyzed (Table 2). In addition,
the communities of the system water and flow equalizer
Bacteria reactors were characterized. To achieve the different culture condi-
The reactors were made of glass in the workshop of tions, two flows were combined in the reactor influent: the
Wageningen University. The reactors had a working volume waste flow containing the fish waste from the flow equalizer
of 3.5 L and were equipped with baffles to improve the and the supplement flow containing the three organic
hydrodynamics (Fig. 2). From the flow equalizer the slurry carbon supplements. In the fourth operation condition,
was continuously pumped into the bacterial culture reactor TAN was added to the supplement flow. The supplements
by a peristaltic pump (Masterflex L/S, Masterflex). The SRT were mixed with distilled water and pumped by a peristaltic
was equal to the HRT as no sludge was returned. Pure pump (PD5001, Heidolph, Germany) into the reactors at a
oxygen was diffused by air-stones to maintain aerobic flow rate which was about 5% of the total flow rate. These
experimental conditions allowed comparing the effects of
Degassing pipe different HRTs, different carbon sources, and different
Inlets for
Oxygen inlet nitrogen sources. Because bacteria prefer TAN over nitrate
acetate
acid
base Waste inlet as a nitrogen source, the effect of those two nitrogen sources
could be investigated. Nitrate was available from the RAS
effluent stream, but it was decreasingly taken up by the
bacteria in the presence of increasing TAN concentrations. A
more detailed description of the experiments is provided in
Schneider (2007) and further in Schneider (2006), Schnei-
der et al. (2006a, b). From the three sampling points (system
water at the fish tanks influent, flow equalizer and bacteria
reactor), samples were siphoned and either analyzed as
Stirrer aqueous samples (50 mL), or sample material was collected
with 2 blades
pH electrode
over time (10.5 L) and centrifuged at 12g for 20 min (Table
Oxygen electrode
2). The supernatant was discarded, and the solid fraction
was collected and freeze dried.
Reactor outlet
Isolation, biochemical and 16S rRNA gene
ribotyping of cultured bacteria
Baffles Aqueous samples (1– 4) were homogenized, and each
homogenate was inoculated on to brain heart infusion
(BHI) agar with 5% sheep blood (home made at CIDC-
Lelystad, The Netherlands), and in parallel on to Cytophaga
agar (Oxoid), and incubated at 22 1C for 5 –7 days. After
Air stone
bacterial growth occurred, morphologically different colo-
Fig. 2. Schematic drawing of the bacteria growth reactor. nies were randomly selected for further typing in a pure
FEMS Microbiol Ecol 60 (2007) 207–219
c 2007 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
4. 210 O. Schneider et al.
Table 2. Sample scheme for the four experimental conditions
Biochemical analysis and DNA isolation and
Sample HRT (h) Sample-ID 16S rRNA gene ribotyping PCR amplification
System water 1 Aqueous sample (50 mL) –
Equalizer 2 Aqueous sample (50 mL) Lyophilized (10.5 L)
Reactor
1.7 g C LÀ1 sodium acetate 7 3 Aqueous sample (50 mL) Lyophilized (10.5 L)
1.7 g C LÀ1 sodium acetate 2 4 Aqueous sample (50 mL) Lyophilized (10.5 L)
2.5 g C LÀ1 molasses 6 5 – Aqueous sample (50 mL)
1.7 g LÀ1 sodium acetate plus 250 mg LÀ1 TAN 6 6 – Aqueous sample (50 mL)
Volumes represent the original sample volume.
plate culture. These were cultured to a monoculture, using denaturation 95 1C for 30 s, annealing at 56 1C for 40 s, and
BHI with 5% sheep blood and identified according to extension at 72 1C for 1 min; and a final extension at 72 1C
standard biochemical tests (Bergey, 1984; Austin Austin, for 5 min. PCR products were verified by electrophoresis on
1987; Barrow Feltham, 1993). If identification was not a 1% (w/v) agarose gel containing ethidium bromide.
possible by these conventional methods, further typing was
carried out using molecular methods, using the Microseq DGGE analysis
500, 16S rRNA gene bacterial identification kits (Applied
Amplicons were separated by DGGE based on the protocol
Biosystems, Foster City, CA), according to the method
of Muyzer Smalla (1998) using the Decode system (Bio-
provided by the manufacturer.
Rad Laboratories, Hercules) with the following modifica-
tions. The polyacrylamide gels consisted of 8% (v/v) poly-
DNA isolation and PCR amplification for acrylamide (ratio of acrylamide to bisacrylamide: 37.5 :1)
molecular characterization of bacterial and 0.5 Â Tris-acetate-EDTA buffer (pH 8.0). Denaturing
communities acrylamide of 100% was defined as 7 M urea and 40%
formamide. The polyacrylamide gels were prepared with
For molecular analyses, DNA was isolated from freeze-dried
denaturing gradients ranging from 30% to 55% to separate
samples with the Fast DNASPIN kit (for soil; QBIOgene,
the generated amplicons of the total bacterial communities.
Cambridge, United Kingdom). Briefly, 0.1 g of freeze-dried
The gels were poured from the top using a gradient maker
material from each sample were placed in Lysing Matrix E
and a pump (Econopump; Bio-Rad) set at a rate of
Tubes with 122 mL of MT buffer and 978 mL of PBS and
4.5 mL minÀ1. Prior to the polymerization of the denaturing
processed three times for 30 s at speed 5.5 m sÀ1. The rest of
gel (gradient volume, 28 mL), a 7.5 mL stacking gel without
the protocol was carried out according to the manufacturer’s
denaturing chemicals was added. Electrophoresis was per-
instructions.
formed first for 5 min at 200 V and then for 16 h at 85 V in
PCR was performed with Taq polymerase kit (Invitrogen,
0.5 Â Tris-acetate-EDTA buffer (pH 8.0) at a constant
Carlsbad, CA) with the universal primer set 0968-a-S-GC-f
temperature of 60 1C. The gels were stained with AgNO3
(5 0 -AACGCGAAGAACCTTA-3 0 ) and S-D-Bact-L1401-a-A-
according to the method of Sanguinetti et al. (1994) and
17 r (5-CGGTGTGTACAAGACCC-3 0 ; N¨ bel et al., 1996),
u
dried overnight at 60 1C. Gels were scanned at 400DPI, and
which amplify the V6 to V8 regions of the bacterial 16S
analyzed with gel analysis software (BIONUMERICS 4.0; Applied
rRNA gene. The first primer has a 40 nucleotide GC rich
Maths BVBA, Sint-Martens-Latem, Belgium).
sequence at the 5 0 end (CGC CGG GGG CGC GCC CCG
GGC GGG GCG GGG GCA CGG GGG G), which allows the
Cloning of the PCR-amplified products
detection of sequence variations of amplified DNA frag-
ments by subsequent denaturing gradient gel electrophoresis 16S rRNA gene-targeted PCR amplicons (1500 bp) were
(DGGE; Muyzer et al., 1993). Each PCR reaction mixture generated with the set of primers 27-f (5-
contained (final volume, 50 mL) 20 mM Tris-HCl (pH 8.4), GTTTGATCCTGGCTCAG-3) and S-D-Bact-1492-a-A-19 r
3 mM MgCl2, each deoxynucleoside triphosphate at a con- (5-CGGCTACCTTGTTACGAC-3; Lane, 1991) and were
centration of 0.2 mM, each primer at a concentration of purified with NucloeSpin Extract II (Macherey-Nagel, The
0.2 mM, 1.25 U of Taq polymerase, and 1 mL of template Netherlands) according to the manufacturer’s instructions.
DNA. Samples were amplified in a Whatman Biometra PCR products were cloned into Escherichia coli XL1-Blue
Thermocycler (G¨ ttingen, Germany) using the following
o competent cells (Stratagene) using the Promega pGEM-T
program: predenaturation at 95 1C for 2 min; 35 cycles of easy vector system (Promega, Madison, WI). Ligation and
c 2007 Federation of European Microbiological Societies FEMS Microbiol Ecol 60 (2007) 207–219
Published by Blackwell Publishing Ltd. All rights reserved
5. Bacteria communities produced on RAS effluents 211
transformation reactions were performed according to the construction of a 16S rRNA gene based phylogenetic tree,
protocol described by the manufacturer. PCR was per- using the neighbour joining method (Saitou Nei, 1987).
formed on cell lysates of ampicillin-resistant transformants Phylogenetic placement was performed in comparison with
using vector specific primers T7 (TAATACGACTCACTA- reference sequences with Jukes–Cantor correction and
TAGG) and Sp6 (GATTTAGGTGACACTATAG) to confirm application of a phylum-level filter as implemented in ARB
the size of the inserts. A total of 96 amplicons of the correct (release February 2005). Chimeric sequences were identified
size (per sample) were subjected to amplified ribosomal by comparison of phylogenetic affiliation of the two respec-
DNA restriction analysis (ARDRA) using the restriction tive 5 0 - and 3- partial sequences. For the tree shown in Fig. 4
enzymes MspI, CfoI, and AluI. From each sample, clones and 5 0 -partial sequences obtained from clones and
corresponding to a unique RFLP pattern were used to sequences determined by ribotyping of cultured isolates
amplify V6–V8 regions of 16S rRNA genes with the primers were used from E. coli position 118 to 412.
968f-GC-f and 1401r as described previously, and they were
selected for subsequent sequence analysis according to their Nucleotide sequence accession numbers
migration position in the DGGE gel compared to the
Partial 16S rRNA gene sequences of the 16S rRNA gene
amplicons of the original DGGE profile of the sample.
clones have been deposited in the GenBank database under
accession numbers DQ788530–DQ788539.
Sequence analysis
PCR amplicons (1.4 kb) of transformants selected by the Results
above-described ARDRA/DGGE screening procedure were
purified with NucloeSpin Extract II (Macherey-Nagel, The Isolation, and biochemical and 16S rRNA gene
Netherlands) according to the manufacturer’s instructions. ribotyping of cultured bacteria
The samples were subjected to DNA sequence analysis
The results from the biochemical and 16S rRNA gene
(BaseClear Lab services, The Netherlands) with the primers
ribotyping for the system water, the equalizer and different
SP6 and T7, yielding two partial sequences (5 0 and 3 0 ) per
reactor broths are given in Table 3. While the system water
clone of c. 500 nucleotides.
and the flow equalizer contained five and seven different
Sequences were analyzed for similarity with sequences
bacteria, only four and three different bacteria were detected
deposited in public databases using the BLAST tool (McGinnis
in the reactor samples.
Madden, 2004) at the National Center for Biotechnology
Information database (http://www.ncbi.nlm.nih.gov/BLAST).
Molecular analysis of bacterial community
Alignment and further phylogenetic analysis of the
structure
sequences were performed using the ARB software package
(Ludwig et al., 2004). The resulting alignments were manu- The phylogenetic affiliations of the clones corresponding to
ally checked and corrected when necessary, and unambigu- prevalent bands in the DGGE sample profile were deter-
ously aligned nucleotide positions were used for mined by sequence analysis (Fig. 3, Table 4, Fig. 4). In the
Table 3. Results from the biochemical and 16S rRNA gene typing for the system water, the equalizer and different reactor broths
System water Equalizer 1.7 g C LÀ1, HRT 7 h 1.7 g C LÀ1, HRT 2 h
% of matching (homology)
Sample ID 1 2 3 4 by ribotyping Method
Bacillus sp. 1 – 1
Edwardsiella sp. 1 99 2
Proteus vulgaris 1 – 1
Aeromonas hydrophilia 1 1 1 – 1
Aeromonas sobria 1 1 – 1
Acinetobacter Iwoffi 1 – 1
Pseudomonas sp. 1 1 – 2
Comamonas sp. 1 99 2
Arcobacter butzlerii sp. 1 1 99 2
Chryseobacterium sp. 1 100 2
Flavobacterium sp. – 1
Myroides sp. 1 1 1 98 and 93 1,2
Sphingobacterium sp. 1 99 2
C, carbon; HRT, hydraulic retention time; method 1, biochemical procedure; method 2, 16S rRNA gene ribotyping.
FEMS Microbiol Ecol 60 (2007) 207–219
c2007 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
6. 212 O. Schneider et al.
1.7g/L–1 sodium acetate, 2.5gC/1 molasses. 1.7gC L–1sodium 1.7gC L–1 sodium
250mg L–1 TAN, 6 h 6h acetate, 7 h acetate, 2 h Equalizer
1
2
3
4
5
6
7
12 8
11
9
10
Fig. 3. 16S rRNA gene-targeted PCR-DGGE analysis of bacterial communities in samples 2–6. Identification of bands was performed by DGGE analysis
of clones. Clones corresponding to bands 1 and 3 were all chimeric, and sequences are therefore not considered.
flow equalizer (sample 2), the predominant bands corre- phylogenetic relations between the detected phylotype and
sponded to sequences most closely related to Sarcina sp., related sequences are displayed in Fig. 4.
Flavobacterium columnare and Catellibacterium terrae
(bands 2, 4, 5). Clones corresponding to bands 1 and 3 were
found to be chimeric, unfortunately prohibiting unambig-
Discussion
uous identification. In sample 3 (1.7 g C LÀ1 sodium acetate, The integrated application of complementary cultivation-
7 h HRT) and in sample 4 (1.7 g C LÀ1 sodium acetate, 2 h dependent and biomolecular approaches allowed for the
HRT), similar profiles were found. For sample 3, DGGE qualitative and semi-quantitative comparison of the bacteria
fingerprinting suggested that the microbial community was communities present in the system water and the flow
dominated by a-proteobacterial populations most closely equalizer, and those that developed in bioreactors operated
related to Rhizobium spp. and Shinella zoogloeoides. In at four different conditions.
sample 4, the most predominant/abundant population was In general, only a limited number of bacterial populations
related to Acinetobacter lwoffi, while this phylotype was less were identified that were common to both the system
abundant in sample 3. Alpha-proteobacterial populations water and the flow equalizer. Examples were Aeromonas sp.
were only detected as minor community components. In and Myroides sp. RAS configuration might have caused
sample 5 (2.5 g C LÀ1 molasses, 6 h HRT), the most abun- such differences in the two bacteria communities. The
dant phylotype was most closely related to Aquaspirillum drumfilter effluent originates from water with a higher
serpens. Comparison of DGGE fingerprints suggested that organic waste load than the tank influent water, which
also a-proteobacterial populations corresponding to bands was treated by the drumfilter. This treatment can reduce
7 and 8 were present, while other minor bands could not be the chemical oxygen demand (COD) load in the system
identified. In sample 6 (1.7 g C LÀ1 sodium acetate, 250 mg water by 50% (own unpublished data). This reduction
TAN/l and 6 h HRT), the main identified components of the affects bacterial numbers, namely by removal of those
microbial community were populations related to Jonesia populations which grow in flocks and on solid particular
spp., Sphaerotilus spp. and Flavobacterium mizutaii. The waste, and of substrates, which are no longer available
c 2007 Federation of European Microbiological Societies FEMS Microbiol Ecol 60 (2007) 207–219
Published by Blackwell Publishing Ltd. All rights reserved
7. Bacteria communities produced on RAS effluents 213
Fig. 4. Phylogenetic tree of bacterial 16S rRNA gene sequences retrieved from the different samples and cultured isolates (^, 16S rRNA gene
ribotyping; , biochemical procedures; V, PCR-DGGE). The tree was constructed from sequences obtained in this study and reference sequences by
neighbor joining procedures, using a bacterial filter, as implemented in ARB (Ludwig et al., 2004). Accession numbers of reference sequences and 16S
rRNA gene clones are provided in the figure. The reference bar indicates 10% sequence divergence.
for bacteria growth. The bacterial strains, found in the all experiments fish were healthy and the system performed
system water and the flow equalizer, included suspected well. The detected bacteria had, therefore, no visible nega-
pathogens at different levels (Tables 3–5). Despite the fact, tive impact on fish health. In general, the bacterial phylo-
however, that potential pathogens were detected, during types found in the system water and flow equalizer are
FEMS Microbiol Ecol 60 (2007) 207–219
c 2007 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
8. 214 O. Schneider et al.
Table 4. Results from the DNA isolation and PCR amplification for the equalizer and different reactor broths
1.7 g C acetate 1.7 g C acetate 2.5 g C LÀ1 molasses, 1.7g C LÀ1, 250 mgTAN LÀ1,
Equalizer LÀ1 HRT 7 h LÀ1 HRT 2 h HRT 6 h HRT 6 h
Sample ID 2 3 4 5 6 Band-ID
Sarcina ventriculi (95) 1 (1) (1) (1) (1) 2
Flavobacterium collumnare (94) 1 (1) (1) (1) (1) 4
Catellibacterium terrae (95) 1 (1) (1) (1) (1) 5
Rhodobacter sphaeroides (93)
Gammaproteobacterium Bioluz (98) (1) 1 (1) (1) 6
Acinetobacter johnsonii (98)
Rhizobium sp. OK-55(97) 1 (1) (1) (1) 7
Rhizobium sp. OK-55(97) 1 (1) 8
Aquaspirillum serpens (98) 1 9
Jonesia denitrificans (91) 1 10
Sphaerotilus sp. IF5 (98) 1 11
Sphingobacterium multivorum (90) 1 12
Named bacteria are the closest match to the analysed sequences; sequence similarity is given in parantheses. Where most closely related sequences
correspond to phylotypes that are not or poorly characterized, the closest cultured reference strain is also provided.
1, identified as present in the sample; (1), presence predicted from identical band migration; C, carbon; HRT, hydraulic retention time.
typical for aquatic, fish farm and wastewater environments was not reported. To grow at a HRT of 2 h, a growth rate of
(Table 5). at least 0.5 hÀ1 is required, which is out of range for
The communities obtained from the reactors operated at Rhizobium spp. at high conductivities. Shorter HRT (e.g.
four different operation conditions were different from the 2 h compared to 7 h) might therefore bear the risk to culture
community of the flow equalizer. Only Arcobacter sp. and mainly potentially pathogenic bacteria.
Myroides sp. were found in both the flow equalizer and in A community similar to that obtained with sodium
one reactor broth samples (sample 3), when communities acetate (sample 3) was found for the reactor using molasses
were analyzed by biochemical and 16S rRNA gene ribotyp- as substrate (sample 5). The major difference was a commu-
ing of culture isolates. Based on comparison of 16S rRNA nity shift from strains represented by bands 7 and 8 to a
gene-targeted PCR-DGGE community fingerprints, all bac- population close to Aquaspirillum serpens, which was not
teria present in the equalizer were also present in the reactor detected as major component in sample 3. Such changes can
broth (Table 4). However, the major community compo- occur if both bacteria utilize similar substrates and can grow
nents in the reactor were composed of other populations, under similar conditions (Table 5). Furthermore, the
which were not found in the equalizer. HRT seemed to have molasses was not sterile. Bacteria other than those existing
a minor effect on the bacterial community as is shown by the in the system might have been introduced through the
results of samples 3 and 4, which differed only in their HRT substrate. Whether the bacteria, close to Aquaspirillum
(7 vs. 2 h). However, in sample 3, a-proteobacterial popula- spp., were more capable to utilize molasses than the Rhizo-
tions close to Rhizobium spp. and Shinella zoogloeoides were bium spp./Shinella zoogloeoides related populations has
the major community components, whereas in sample 4 (2 h nevertheless not been reported elsewhere.
HRT), a population most closely related to Acinetobacter When TAN was applied in addition to sodium acetate, the
spp. was the major component. This suggests that the bacteria community changed significantly (samples 3 and
previously dominating a-proteobacterial populations were 6). Nearly all bacteria, which were detected in sample 3 were
out-competed at this low HRT. This corroborates data from also present in sample 6, but another three were also found
Singleton et al. (1982), who reported growth rates for in sample 6. These bacteria were close to Sphaerotilus spp.,
Rhizobium spp. as 0.7–0.2 and 0.4–0.2 hÀ1 for water Sphingobacterium/Flavobacterium spp. and Jonesia spp. (Fig.
conductivities of 1200 and 6000 mS cmÀ1, respectively. 4). Pathogenicity has not been reported for any of these
The experimental conditions were in this range strains (Table 5). Sphingobacterium spp. grow well on swine
(2000–3000 mS cmÀ1). In contrast, Acinetobacter spp. grown manure, where TAN is a major nitrogen source (Leung
on sodium acetate have higher growth rates of 0.2–0.8 hÀ1 at Topp, 2001). Furthermore, Sphaerotilus- and Jonesia spp.-
25 1C compared to the high conductivity conditions related populations have been found in wastewater and mud
(Oerther et al., 2002). Unfortunately water conductivity (Table 5). All three might be then superior to other,
c 2007 Federation of European Microbiological Societies FEMS Microbiol Ecol 60 (2007) 207–219
Published by Blackwell Publishing Ltd. All rights reserved
9. Bacteria communities produced on RAS effluents 215
Table 5. Habitat and growth conditions and pathogenicity for bacterial phylotypes or their closest related strains found in the different samples
Habitat and growth Pathogenicity focussing
Bacteria conditions on animals and fish Reference
Bacillus sp. Saprophytic Some strains, f.i. B. cereus (in carp and Weber (1997); Austin Austin
Waste water, paper mill slime striped bass), B. mycoides (in channel (1999); Oppong et al. (2003);
catfish), and B. subtilis (in carp) Tchobanoglous et al. (2003)
Edwardsiella sp. 23–28 1C Some fish pathogenic enterobacteria: Austin Austin (1987); Abbott
Aquatic habitats and especially fish, E.tarda (eel), E.ictaluri (channel Janda (2001)
amphibians, reptiles, and birds catfish), different effects on various
species, reaching from fatal to none
Proteus vulgaris Saprophytic Only few indication Austin Austin (1987); Weber
Soil, water, integral part of gut flora (1997); Manos Belas (2001);
Tanaka R et al. (2004)
Aeromonas hydrophilia Facultative anaerobic, 4–37 1C Facultative opportunistic found as Kinne (1984); Rice et al. (1984);
Different salinities well on healthy fish Austin Austin (1987); Meyer-Reil
Aquatic habitats, waste water Koester (1993); Weber (1997);
found frequently at fish farms Leonard et al. (2000)
Aeromonas sobria Facultative anaerobic 4–37 1C Facultative opportunistic or not Kinne (1984); Austin Austin (1987);
Different salinities necessarily attributed as pathogenic Meyer-Reil Koester (1993); Weber
Aquatic habitats, waste water found as well on healthy fish (1997)
frequently on fish farms
Acinetobacter iwoffi Aerobic 20–30 1C Facultative opportunistic, few Rice et al. (1984); Austin Austin
Different salinities indications (1987); Meyer-Reil Koester (1993);
Soil, aquatic habitats, waste water Fang et al. (2002); Wagner Loy
frequently on fish farms (2002)
Pseudomonas sp. Mesophilic temperatures Some facultative opportunistic, or Adamse (1968a); Austin Austin
Different salinities pathogenic strains reported: f.i. P. (1999); Palleroni et al. (1999)
Soils, water, sewage, animals, anguilliseptica (in eel, sea bream
plants and sea bass)
Sphaerotilus natans Aerobic/anaerobic Not reported Adamse (1968b); Pasveer (1968);
Freshwater Spring (2002); Schonborn (2003)
Sludges, waste water
Comamonas sp. Aerobic Rare opportunistic pathogens, no Etchebehere et al. (2001); Gumaelius
20–37 1C evidence of pathogenic effect on et al. (2001); Willems de Vos (2002)
Waste water, activated sludge, healthy people
animals’ blood
Aquaspirillum serpens Aerobic Not reported Payne (1981); Pot et al. (1999); Tal
Different salinities et al. (2003); Thomsen et al. (2004)
Denitrifcication reactors as well in
marine recirculation systems
Rhizobium/ Facultative aerobic Not reported Payne (1981); O’Hara Daniel (1985);
Mesorhizobium Soil, denitrification reactors, Batut Boistard (1994); Encarnacion
culturable on wastewater sludge, et al. (1995); Sadowsky Graham
aquatic (2000); Rebah et al. (2001);
systems, denitrification reactors Etchebehere et al. (2002); Liu et al.
(2005)
Shinella zoogloeoides Aerobic Not reported Kargi Karapinar (1995); Dugan et al.
Aquatic systems, domestic sewage (1999)
and aerobic sewage-treatment
systems
Castellibacterium sp./ Fresh to salt water Not reported Kersters et al. (2003); Tanaka Y et al.
Rhodobacter sp. Activated sludge, marine sludge (2004); Cytryn et al. (2005a, b)
Arcobacter butzlerii/sp. Aerobic Possibly involved Moreno et al. (2003); Tanaka R et al.
15–37 1C (2004); Lehner et al. (2005)
Gut microbiota, surface and
ground waters
Seawage and activated sludge
FEMS Microbiol Ecol 60 (2007) 207–219
c2007 Federation of European Microbiological Societies
Published by Blackwell Publishing Ltd. All rights reserved
10. 216 O. Schneider et al.
Table 5. Continued.
Habitat and growth Pathogenicity focussing
Bacteria conditions on animals and fish Reference
Chryseobacterium sp. Aerobic Pathogenic, f.i. C. scophthalmum (in Urdaci et al. (1998); Austin Austin
Different salinities turbot), C. balustinum (in marine fish) (1999); Jooste Hugo (1999);
Soil, plants, aquatic habitat, Bernardet Nakagawa (2000);
activated sludge Mustafa et al. (2002);
Bernardet (2005)
Flavobacterium sp. Aerobic Facultative, mostly found externally, Kinne (1984); Austin Austin (1987);
5–42 1C may induce skin necrosis after stress Murray et al. (1990); Meyer-Reil
Salinity below 1% found as well on healthy fish, some Koester (1993); Bernardet
Soil, aquatic habitat frequently species are very pathogenic Nakagawa (2000); Bernardet et al.
at fish farms (2005)
Myroides sp. Aerobic Opportunistic Gonzalez et al. (2000); Hugo et al.
25–30 1C (2000)
Human instestine, soil, water
Sphingobacterium sp. Aerobic Not reported Holmes (1999); Leung Topp (2001);
Soil, activated sludge, gut fauna, Tanaka R et al. (2004)
liquid swine manure
Sarcina ventriculi Obligate anaerobic, but not Not reported Goodwin Zeikus (1987); Jung et al.
oxygen sensitive (1993); Snell-Castro et al. (2005)
30–37 1C
Gut fauna
Jonesia quinghaiensis Aerobic Not reported Schumann et al. (2004)
20–30 1C
Different salinities
Mud
outcompeted populations in the utilization of TAN, result- ing community were not identified. Those bacteria have
ing in higher growth rates. been found in other studies, focusing on the system as a
Given the pathogenic risk associated with short HRTs, it whole by investigating its components (Tal et al., 2003;
is advisable to choose for HRTs of 6–7 h. The choice of Cytryn et al., 2005). Investigations of heterotrophic bacteria
organic C donor seems of less importance, as the obtained communities yielded some similar results, such as the
communities in the presence of sodium acetate or molasses, detection of Pseudomonas spp., Aeromonas spp., Aquaspir-
respectively, did not change in their pathogenicity. More- illum spp. and others (Leonard et al., 2000; Tal et al., 2003).
over, the addition of TAN did not increase the risk of Anyway, it is unlikely to find completely identical bacterial
potentially pathogenic populations, as revealed by the communities in RAS, because of differences in their envir-
comparison of samples 3 and 6. Two considerations have to onmental conditions (marine vs. freshwater), configurations
be made. The ‘native’ nitrogen source supplied in the RAS (e.g. presence of UV, foam fractionators), and in the
effluent stream is nitrate. To utilize this nitrogen species, the cultured animals.
system design did not change and the reactor can easily be
installed after the drumfilter. If TAN should be used, the
system would have to be modified to eliminate nitrification.
Conclusion
The only advantage to using TAN might then be the The bacteria community found in the system water and in
potentially higher nutritional value of the obtained bacteria the flow equalizer contained some possible opportunistic
biomass. This advantage would have to be confirmed by pathogens, but did not result in severe disease symptoms or
additional experiments. Generally, the pathogenic risk and production losses during the fish culture operation. The
nutritional value of all obtained bacterial material has to be bacteria community of the flow equalizer was semi-quantita-
further investigated in feeding trials, if the bacteria biomass tively different from the communities found in the bacteria
should be used as aquatic feed. reactors. However, all major community components were
To compare the occurrence of bacteria found in the present in both equalizer slurry and reactor broths. Hydrau-
system water, the flow equalizer and in the bacteria reactor lic retention times (7 h vs. 2 h) influenced the bacteria
with bacteria found in RAS in general is difficult, because community composition, resulting in a more abundant
literature data is scarce. Because no biofilter material was fraction of potentially pathogenic populations related to
investigated in this study, bacteria belonging to the nitrify- Acinetobacter at 2 h HRT compared to 7 h HRT. At 7 h
c 2007 Federation of European Microbiological Societies FEMS Microbiol Ecol 60 (2007) 207–219
Published by Blackwell Publishing Ltd. All rights reserved
11. Bacteria communities produced on RAS effluents 217
bacteria close to Rhizobium spp. and Shinella zoogloeoides Bernardet JF, Vancanneyt M, Matte-Tailliez O, Grisez L, Tailliez P,
formed the major components of the community. The use of Bizet C, Nowakowski M, Kerouault B Swings J (2005)
molasses instead of sodium acetate caused a shift in compo- Polyphasic study of Chryseobacterium strains isolated from
sition to a bacterial community dominated by a population diseased aquatic animals. Syst Appl Microbiol 28: 640–660.
similar to Aquaspirillum serpens. Providing TAN in addition Burford MA, Thompson PJ, McIntosh RP, Bauman RH
to nitrate as nitrogenous substrate led to the occurrence of Pearson DC (2003) Nutrient and microbial dynamics in high-
bacteria close to Sphaerotilus spp., Flavobacterium mizutaii intensity, zero-exchange shrimp ponds in Belize. Aquaculture
and Jonesia spp. It was concluded from those results that a 219: 393–411.
reactor operation regime of 6–7 h HRT is recommended, and Chen SL, Coffin DE Malone RF (1997) Sludge production and
that the type of substrate (sodium acetate or molasses, TAN management for recirculating aquacultural systems. J World
or nitrate) is less important. Considering conventional RAS Aquac Soc 28: 303–315.
configurations, nitrate might be preferred over TAN. How- Cytryn E, Minz D, Gelfand I, Neori A, Gieseke A, De Beer D
ever, for all the obtained bacteria communities, additional Van Rijn J (2005a) Sulfide-oxidizing activity and bacterial
tests are required to investigate their pathogenic risk and community structure in a fluidized bed reactor from a zero-
nutritional values as aquatic feed in more detail. discharge mariculture system. Environ Sci Technol 39:
1802–1810.
Cytryn E, van Rijn J, Schramm A, Gieseke A, de Beer D Minz D
Acknowledgements (2005b) Identification of bacteria potentially responsible for
oxic and anoxic sulfide oxidation in biofilters of a recirculating
The authors would like to thank the hatchery and the
mariculture system. Appl Environ Microbiol 71: 6134–6141.
laboratory staff of the AFI group for their support during Dugan PR, Stoner DL Pickrum HM (1999) The genus Zooglea.
the study. This study was financed by the European Union in The Prokaryotes: An Evolving Electronic Resource for the
the frame of the ZAFIRA project (ICA4-CT-2001-10025). Microbiological Community (Dworkin M, Falkow S, Rosenberg
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