PCR-DGGE analysis of denitrifying bacteria in a metallurgic wastewater treatment process

2002 ◽  
Vol 46 (1-2) ◽  
pp. 333-336 ◽  
Author(s):  
N. Noda ◽  
S. Yoshie ◽  
T. Miyano ◽  
S. Tsuneda ◽  
A. Hirata ◽  
...  
Keyword(s):  
Fluidized Bed ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Precious Metals ◽  
Packed Bed ◽  
Population Diversity ◽  
Operating Conditions ◽  
Industrial Wastes ◽  
Wastewater Treatment Process ◽  
Dgge Analysis ◽  
Gradient Gel Electrophoresis

The wastewater generated from the processes of recovering precious metals from industrial wastes contains high concentrations of acids such as nitric acid and of salts. Biological nitrogen removal from this wastewater was attempted by using a circulating bioreactor system equipped with an anoxic packed bed or an anoxic fluidized bed and an aerobic three-phase fluidized bed. The system was found to effectively remove nitrogen from the diluted wastewater (T-N; 1,000–4,000 mg litre−1). The microbial population structure of activated sludge in an anoxic reactor was analyzed by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S ribosomal DNA (rDNA) fragments. DGGE analysis under different operating conditions demonstrated the presence of some distinguishable bands in the separation pattern, which were most likely derived from many different species constituting the microbial communities. Furthermore, the population diversity varied in accordance with the nitrate-loading rate, water temperature and reactor condition. Some major DGGE bands were excised, reamplified and directly sequenced. It was revealed that the dominant population in the anoxic reactor were affiliated with the β subclass of the class Proteobacteria.

Characterization of microbial community in nitrogen removal process of metallurgic wastewater by PCR-DGGE

2002 ◽  
Vol 46 (11-12) ◽  
pp. 93-98 ◽  
Author(s):  
S. Yoshie ◽  
N. Noda ◽  
T. Miyano ◽  
S. Tsuneda ◽  
A. Hirata ◽  
...  
Keyword(s):  
Microbial Community ◽  
Nitrogen Removal ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Precious Metals ◽  
Community Analysis ◽  
Packed Bed ◽  
Community Diversity ◽  
Industrial Wastes ◽  
Microbial Community Analysis ◽  
Gradient Gel Electrophoresis

The metallurgic wastewater generated from the processes of recovering precious metals from industrial wastes contains high concentrations of nitrogen compounds such as ammonia and nitric acid and of salts such as sodium chloride and sodium sulfate. Biological nitrogen removal from this wastewater was attempted by a circulating bioreactor system equipped with an anoxic packed bed and an aerobic fluidized bed. The anoxic packed bed of this system was found to effectively remove nitrite and nitrate from the wastewater by denitrification at a removal ratio of 97%. As a result of denitrification activity tests at various NaCl concentrations, the sludge obtained from the anoxic packed bed exhibited accumulation of nitrite at 5.0 and 8.4% NaCl concentrations, suggesting that the reduction of nitrite is the key step in the denitrification pathway under hypersaline conditions. The microbial community analysis by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S ribosomal DNA (rDNA) fragments revealed that the community diversity varied in accordance with water temperature, nitrate-loading rate and ionic strength. When particular major DGGE bands were excised, reamplified and directly sequenced, the dominant species in the anoxic packed bed were affiliated with the beta and gamma subclasses of the class Proteobacteria such as Alcaligenes defragrans and Pseudomonas spp., respectively.

Nutrients removal in hybrid fluidised bed bioreactors operated with aeration cycles

2007 ◽  
Vol 55 (8-9) ◽  
pp. 51-58 ◽  
Author(s):  
M.A. Martín Martín ◽  
L. López Enríquez ◽  
M. Fernández-Polanco ◽  
S. Villaverde ◽  
P.A. García-Encina
Keyword(s):  
Total Nitrogen ◽  
Industrial Wastewater ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Operating Conditions ◽  
Synthetic Wastewater ◽  
Fluidised Bed ◽  
Total N ◽  
Biological Communities ◽  
Gradient Gel Electrophoresis ◽  
Polymerase Chain

Two hybrid fluidised bed reactors filled with sepiolite and granular activated carbon (GAC) were operated with short cycled aeration for removing organic matter, total nitrogen and phosphorous, respectively. Both reactors were continuously operated with synthetic and/or industrial wastewater containing 350–500 mg COD/L, 110–130 mg NKT/L, 90–100 mg NH3-N/L and 12–15 mg P/L for 8 months. The reactor filled with sepiolite, treating only synthetic wastewater, removed COD, ammonia, total nitrogen and phosphorous up to 88, 91, 55 and 80% with a hydraulic retention time (HRT) of 10 h, respectively. These efficiencies correspond to removal rates of 0.95 kgCODm−3d−1 and 0.16 kg total N m−3d−1.The reactor filled with GAC was operated for 4 months with synthetic wastewater and 4 months with industrial wastewater, removing 98% of COD, 96% of ammonia, and 66% of total nitrogen, with an HRT of 13.6 h. No significant phosphorous removing activity was observed in this reactor. Microbial communities growing with both reactors were followed using polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) techniques. The microbial fingerprints, i.e. DGGE profiles, indicated that biological communities in both reactors were stable along the operational period even when the operating conditions were changed.

scholarly journals Biodiversity of Thermophilic Prokaryotes with Hydrolytic Activities in Hot Springs of Uzon Caldera, Kamchatka (Russia)

2008 ◽  
Vol 75 (1) ◽  
pp. 286-291 ◽  
Author(s):  
Ilya V. Kublanov ◽  
Anna A. Perevalova ◽  
Galina B. Slobodkina ◽  
Aleksander V. Lebedinsky ◽  
Salima K. Bidzhieva ◽  
...  
Keyword(s):  
Hot Springs ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Rrna Gene ◽  
Specific Primers ◽  
Uzon Caldera ◽  
Dgge Analysis ◽  
Hydrolytic Activities ◽  
Gradient Gel Electrophoresis ◽  
Polymeric Substrates

ABSTRACT Samples of water from the hot springs of Uzon Caldera with temperatures from 68 to 87�C and pHs of 4.1 to 7.0, supplemented with proteinaceous (albumin, casein, or α- or β-keratin) or carbohydrate (cellulose, carboxymethyl cellulose, chitin, or agarose) biological polymers, were filled with thermal water and incubated at the same sites, with the contents of the tubes freely accessible to the hydrothermal fluid. As a result, several enrichment cultures growing in situ on different polymeric substrates were obtained. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments obtained after PCR with Bacteria-specific primers showed that the bacterial communities developing on carbohydrates included the genera Caldicellulosiruptor and Dictyoglomus and that those developing on proteins contained members of the Thermotogales order. DGGE analysis performed after PCR with Archaea- and Crenarchaeota-specific primers showed that archaea related to uncultured environmental clones, particularly those of the Crenarchaeota phylum, were present in both carbohydrate- and protein-degrading communities. Five isolates obtained from in situ enrichments or corresponding natural samples of water and sediments represented the bacterial genera Dictyoglomus and Caldanaerobacter as well as new archaea of the Crenarchaeota phylum. Thus, in situ enrichment and consequent isolation showed the diversity of thermophilic prokaryotes competing for biopolymers in microbial communities of terrestrial hot springs.

PCR-DGGE Analysis of Bacterial Diversity of Intestinal System in Hyperlipidemia Rats

2013 ◽  
Vol 726-731 ◽  
pp. 898-901
Author(s):  
Ri Na Wu ◽  
Xiao Meng Pang ◽  
Xi Yan Wang ◽  
Jun Rui Wu
Keyword(s):  
Bacterial Diversity ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Intestinal Flora ◽  
Intestinal Bacteria ◽  
Control Group ◽  
Rrna Gene ◽  
Dgge Analysis ◽  
Gradient Gel Electrophoresis ◽  
The Relationship ◽  
Insight Into

Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene has been regarded as one of powerful tools for gaining insight into the bacterial diversity of intestinal system. In the present study, hyperlipidemia model was constructed in rat according to the tests of blood lipids. Fecal samples of rats were collected after 60d feeding, and DGGE was used to investigate the diversities of intestinal bacteria in the artificially-induced hyperlipidemia rats and normal rats. The results showed that two patterns had similarities, but there were also some different bacteria communities. Moreover, control group had much more bands than model group on gel, showing species in intestinal of model rats might be deduced by hyperlipidemia. It will be helpful to explore the relationship between hyperlipidemia and intestinal flora.

scholarly journals Identification of Bacteria Potentially Responsible for Oxic and Anoxic Sulfide Oxidation in Biofilters of a Recirculating Mariculture System

2005 ◽  
Vol 71 (10) ◽  
pp. 6134-6141 ◽  
Author(s):  
Eddie Cytryn ◽  
Jaap van Rijn ◽  
Andreas Schramm ◽  
Armin Gieseke ◽  
Dirk de Beer ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Sulfide Oxidation ◽  
16S Rrna Genes ◽  
Microbial Consortia ◽  
Rrna Genes ◽  
Marine Aquaculture ◽  
Dgge Analysis ◽  
Dna And Rna ◽  
Gradient Gel Electrophoresis

ABSTRACT Bacteria presumably involved in oxygen- or nitrate-dependent sulfide oxidation in the biofilters of a recirculating marine aquaculture system were identified using a new application of reverse transcription-PCR denaturing gradient gel electrophoresis (DGGE) analysis termed differential-transcription (DT)-DGGE. Biofilter samples were incubated in various concentrations of sulfide or thiosulfate (0 to 5 mM) with either oxygen or nitrate as the sole electron acceptor. Before and after short-term incubations (10 to 20 h), total DNA and RNA were extracted, and a 550-bp fragment of the 16S rRNA genes was PCR amplified either directly or after reverse transcription. DGGE analysis of DNA showed no significant change of the original microbial consortia upon incubation. In contrast, DGGE of cDNA revealed several phylotypes whose relative band intensities markedly increased or decreased in response to certain incubation conditions, indicating enhanced or suppressed rRNA transcription and thus implying metabolic activity under these conditions. Specifically, species of the gammaproteobacterial genus Thiomicrospira and phylotypes related to symbiotic sulfide oxidizers could be linked to oxygen-dependent sulfide oxidation, while members of the Rhodobacteraceae (genera Roseobacter, Rhodobacter, and Rhodobium) were putatively active in anoxic, nitrate-dependent sulfide oxidation. For all these organisms, the physiology of their closest cultured relatives matches their DT-DGGE-inferred function. In addition, higher band intensities following exposure to 5 mM sulfide and nitrate were observed for Thauera-, Hydrogenophaga-, and Dethiosulfovibrio-like phylotypes. For these genera, nitrate-dependent sulfide oxidation has not been documented previously and therefore DT-DGGE might indicate a higher relative tolerance to high sulfide concentrations than that of other community members. We anticipate that DT-DGGE will be of general use in tracing functionally equivalent yet phylogenetically diverse microbial populations in nature.

scholarly journals Nutrient Amendments in Soil DNA Stable Isotope Probing Experiments Reduce the Observed Methanotroph Diversity

2006 ◽  
Vol 73 (3) ◽  
pp. 798-807 ◽  
Author(s):  
Aur�lie C�bron ◽  
Levente Bodrossy ◽  
Nancy Stralis-Pavese ◽  
Andrew C. Singer ◽  
Ian P. Thompson ◽  
...  
Keyword(s):  
Stable Isotope ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Stable Isotope Probing ◽  
Population Diversity ◽  
Type I ◽  
Mineral Salts ◽  
Bacterial Populations ◽  
Soil Dna ◽  
Gradient Gel Electrophoresis ◽  
Soil Water Conditions

ABSTRACT Stable isotope probing (SIP) can be used to analyze the active bacterial populations involved in a process by incorporating 13C-labeled substrate into cellular components such as DNA. Relatively long incubation times are often used with laboratory microcosms in order to incorporate sufficient 13C into the DNA of the target organisms. Addition of nutrients can be used to accelerate the processes. However, unnatural concentrations of nutrients may artificially change bacterial diversity and activity. In this study, methanotroph activity and diversity in soil was examined during the consumption of 13CH4 with three DNA-SIP experiments, using microcosms with natural field soil water conditions, the addition of water, and the addition of mineral salts solution. Methanotroph population diversity was studied by targeting 16S rRNA and pmoA genes. Clone library analyses, denaturing gradient gel electrophoresis fingerprinting, and pmoA microarray hybridization analyses were carried out. Most methanotroph diversity (type I and type II methanotrophs) was observed in nonamended SIP microcosms. Although this treatment probably best reflected the in situ environmental conditions, one major disadvantage of this incubation was that the incorporation of 13CH4 was slow and some cross-feeding of 13C occurred, thereby leading to labeling of nonmethanotroph microorganisms. Conversely, microcosms supplemented with mineral salts medium exhibited rapid consumption of 13CH4, resulting in the labeling of a less diverse population of only type I methanotrophs. DNA-SIP incubations using water-amended microcosms yielded faster incorporation of 13C into active methanotrophs while avoiding the cross-feeding of 13C.

Biodegradation of MTBE and BTEX in an aerobic fluidized bed reactor

2003 ◽  
Vol 47 (9) ◽  
pp. 123-128 ◽  
Author(s):  
A. Pruden ◽  
M. Sedran ◽  
M. Suidan ◽  
A. Venosa
Keyword(s):  
Fluidized Bed ◽  
Flow Rate ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Oxygen Demand ◽  
Fluidized Bed Reactor ◽  
Methyl Tert Butyl Ether ◽  
Reactor Performance ◽  
Butyl Ether ◽  
Acclimation Period ◽  
Gradient Gel Electrophoresis

An aerobic fluidized bed reactor (FBR) was operated for the removal of methyl tert-butyl ether (MTBE) and benzene, toluene, ethylbenzene, and p-xylene (BTEX) from water. The reactor was seeded with a mixed culture adapted to MTBE. Granular activated carbon (GAC) was used as the biological attachment medium. Influent MTBE to the reactor was 7.8 mg/L MTBE, with a flow rate of 22.7 L/day, and an empty bed contact time of 1 hour. The acclimation period required was relatively short, about 30 days before reaching an average stable effluent concentration of 18.5 ± 10 μg/L. BTEX was introduced to the feed at an equivalent chemical oxygen demand (COD) as the MTBE at day 225 and was biodegraded spontaneously with no apparent acclimation period required. The average influent of each of the four BTEX compounds was about 2 mg/L, and the range of the average effluent concentrations was 1.4-2.2 μg/L. After achieving 180 days of stable performance with BTEX addition, the total flow rate to the reactor was gradually increased by 20% increments to 160% of the original flow (36.4 L/day). Increases by 20% and 40% had no apparent effect on reactor performance, but increase by 60% required 30 days before effluent quality returned to previous values. Composition of the culture was monitored throughout operation of the reactor using denaturing gradient gel electrophoresis (DGGE). The culture consisted of Flavobacteria-Cytophaga and organisms with high similarity to the known MTBE degrader PM1.

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