Characterization of the microbial diversity in a biotreatment process using non-culture based methods

2000 ◽  
Vol 42 (3-4) ◽  
pp. 143-148 ◽  
Author(s):  
S.J. You ◽  
C.F. Ouyang ◽  
S.F. Lin ◽  
W.T. Liu
Keyword(s):  
Activated Sludge ◽  
Microbial Population ◽  
Molecular Techniques ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Biological Nutrient Removal ◽  
Growth State ◽  
Microbial Structures ◽  
Population Structures

Molecular techniques were used to compare the microbial community of suspended activated sludge and attached biofilm in a biological nutrient removal process (TNCU-I) operated under various COD/total P (COD/TP) feed ratios. Analysis of restriction fragment length polymorphism of amplified 16S rRNA genes indicated that the microbial population structures were more closely related among those sludge samples taken from aerobic sludge than that taken from biofilm attached on the RBC. The use of different COD/TP feed ratios (300/2.5-300/25) had no significant effect on the change of microbial structures of sludge samples. The 16S rDNA clone library further indicated that at least eight and six different microbial populations were present in activated sludge and RBC biofilm, respectively. The phylogenetic analysis revealed that six of those eight major clones obtained from the suspended activated sludge were from the beta-subclass of the class Proteobacteria. In contrast, only one clone obtained from biofilm belonged to the beta-subclass of the Proteobacteria. This difference in the microbial population structure was possibly attributed to the growth state (suspended or attached) or carbon source (autographs or heterotrophs) of the sludge samples rather than the effect of the COD/TP ratio used.

scholarly journals Coaggregation among Nonflocculating Bacteria Isolated from Activated Sludge

2003 ◽  
Vol 69 (10) ◽  
pp. 6056-6063 ◽  
Author(s):  
Anushree Malik ◽  
Masashi Sakamoto ◽  
Shohei Hanazaki ◽  
Masamitsu Osawa ◽  
Takanori Suzuki ◽  
...  
Keyword(s):  
16S Rrna ◽  
Activated Sludge ◽  
Aggregate Size ◽  
Spectrophotometric Assay ◽  
Full Length ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Acinetobacter Junii ◽  
Acinetobacter Johnsonii

ABSTRACT Thirty-two strains of nonflocculating bacteria isolated from sewage-activated sludge were tested by a spectrophotometric assay for their ability to coaggregate with one other in two-membered systems. Among these strains, eight showed significant (74 to 99%) coaggregation with Acinetobacter johnsonii S35 while only four strains coaggregated, to a lesser extent (43 to 65%), with Acinetobacter junii S33. The extent and pattern of coaggregation as well as the aggregate size showed good correlation with cellular characteristics of the coaggregating partners. These strains were identified by sequencing of full-length 16S rRNA genes. A. johnsonii S35 could coaggregate with strains of several genera, such as Oligotropha carboxidovorans, Microbacterium esteraromaticum, and Xanthomonas spp. The role of Acinetobacter isolates as bridging organisms in multigeneric coaggregates is indicated. This investigation revealed the role of much-neglected nonflocculating bacteria in floc formation in activated sludge.

Prokaryote community dynamics in anaerobic co-digestion of swine manure, rice straw and industrial clay residuals

2016 ◽  
Vol 74 (4) ◽  
pp. 824-835 ◽  
Author(s):  
Janet Jiménez ◽  
Susanne Theuerl ◽  
Ingo Bergmann ◽  
Michael Klocke ◽  
Gilda Guerra ◽  
...  
Keyword(s):  
Rice Straw ◽  
Community Dynamics ◽  
Swine Manure ◽  
Molecular Techniques ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Nutritional Conditions ◽  
Continuously Stirred Tank Reactor ◽  
Positive Effect ◽  
Methanosarcina Species

The aim of this study was to analyze the effect of the addition of rice straw and clay residuals on the prokaryote methane-producing community structure in a semi-continuously stirred tank reactor fed with swine manure. Molecular techniques, including terminal restriction fragment length polymorphism and a comparative nucleotide sequence analyses of the prokaryotic 16S rRNA genes, were performed. The results showed a positive effect of clay addition on methane yield during the co-digestion of swine manure and rice straw. At the digestion of swine manure, the bacterial phylum Firmicutes and the archaeal family Methanosarcinaceae, particularly Methanosarcina species, were predominant. During the co-digestion of swine manure and rice straw the microbial community changed, and with the addition of clay residual, the phylum Bacteroidetes predominated. The new nutritional conditions resulted in a shift in the archaeal family Methanosarcinaceae community as acetoclastic Methanosaeta species became dominant.

scholarly journals 1358. A Novel Rapidly Growing Mycobacteria (RGM) Species Causing Soft Tissue and Orthopedic Hardware Infection After Trauma

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S492-S492
Author(s):  
David C Nguyen ◽  
Michelle Lisgaris ◽  
Sruthi Vasireddy ◽  
Richard J Wallace ◽  
Federico Perez ◽  
...  
Keyword(s):  
Soft Tissue ◽  
16S Rrna ◽  
Antibiotic Susceptibility ◽  
Molecular Techniques ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Mycobacterium Fortuitum ◽  
Rapidly Growing Mycobacteria ◽  
Maldi Tof

Abstract Background The widespread use of molecular techniques has resulted in increasing numbers of newly characterized rapidly growing mycobacteria (RGM). Many RGM cause soft tissue and orthopedic hardware infection, particularly after trauma. RGM species identification remains challenging with few genetic differences between species. Methods We describe a case involving RGM. We report results of matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (Bruker Biotyper), sequencing of rpoB, erm(39), and 16S rRNA genes, and antibiotic susceptibility testing (AST). We review previous reports describing similar RGM infections. Results A 58-year-old male sustained multiple fractures and right thigh compartment syndrome after a motorcycle accident. He underwent fasciotomy and multi-stage surgical fixations. 3 months later, he had wound dehiscence, purulence and multiple fluid collections of his right leg and knee requiring surgical drainage and removal of orthopedic hardware. After 4 days, acid-fast bacilli grew on routine bacterial culture media. MALDI-TOF identified the isolate as Mycobacterium mageritense. In contrast, sequencing of 16S rRNA (100% identity) and erm(39) (> 99% identity) identified the isolate as Mycobacterium houstonense; erm(39) only had 80% similarity with Mycobacterium fortuitum. Sequencing of rpoB showed a 19 bp difference with the M. houstonense type strain, and showed similarity to M. fortuitum (97.64%) than M. houstonense (97.45%). AST demonstrated resistance to clarithromycin only. After initial treatment with imipenem, ciprofloxacin, and doxycycline, definite therapy with ciprofloxacin and doxycycline was successful. In the literature, we found one case each of M. mageritense and M. houstonense infection after trauma. Conclusion This case highlights the importance of RGM other than M. fortuitum as a cause of soft tissue and orthopedic hardware infections, and illustrates the difficulty of identifying them to the species level. Sequencing of erm(39) and 16S rRNA gene identified the isolate as M. houstonense, but the larger difference (>2.5%) in rpoB sequence suggests a novel species. Further characterization is underway. Efforts to determine RGM species and antibiotic susceptibility give important insight into diagnosis and management. Disclosures All authors: No reported disclosures.

Phenotypic and molecular characterization of chickpea rhizobia isolated from different areas of Tunisia

2007 ◽  
Vol 53 (3) ◽  
pp. 427-434 ◽  
Author(s):  
Boulbaba L’taief ◽  
Bouaziz Sifi ◽  
Maher Gtari ◽  
Mainassara Zaman-Allah ◽  
Mokhtar Lachaâl
Keyword(s):  
Rflp Analysis ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Phenotypic Characteristics ◽  
Cicer Arietinum L ◽  
Mesorhizobium Ciceri ◽  
Length Polymorphisms ◽  
Reference Strains ◽  
Tolerance To Salinity

Several phenotypic markers were used in this study to determine the biodiversity of rhizobial strains nodulating Cicer arietinum L. in various areas of Tunisia. They include symbiotic traits, the use of 21 biochemical substrates, and tolerance to salinity and pH. In addition, restriction fragment length polymorphisms (RFLPs) of PCR-amplified 16S rDNA were compared with those of reference strains. Numeric analysis of the phenotypic characteristics showed that the 48 strains studied fell into three distinct groups. This heterogeneity was highly supported by the RFLP analysis of 16S rRNA genes, and two ribotypes were identified. Chickpea rhizobia isolated from Tunisian soils are both phenotypically and genetically diverse. Results showed that 40 and 8 isolates were assigned, respectively, to Mesorhizobium ciceri and Mesorhizobium mediterraneum .

scholarly journals Bioaugmentation of Activated Sludge by an Indigenous 3-Chloroaniline-Degrading Comamonas testosteroni Strain, I2gfp

2000 ◽  
Vol 66 (7) ◽  
pp. 2906-2913 ◽  
Author(s):  
Nico Boon ◽  
Johan Goris ◽  
Paul De Vos ◽  
Willy Verstraete ◽  
Eva M. Top
Keyword(s):  
Community Structure ◽  
Microbial Community ◽  
Activated Sludge ◽  
Microbial Community Structure ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Dynamic Change ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Adaptation Period ◽  
Comamonas Testosteroni

ABSTRACT A strain identified as Comamonas testosteroni I2 was isolated from activated sludge and found to be able to mineralize 3-chloroaniline (3-CA). During the mineralization, a yellow intermediate accumulated temporarily, due to the distalmeta-cleavage of chlorocatechol. This strain was tested for its ability to clean wastewater containing 3-CA upon inoculation into activated sludge. To monitor its survival, the strain was chromosomally marked with the gfp gene and designated I2gfp. After inoculation into a lab-scale semicontinuous activated-sludge (SCAS) system, the inoculated strain maintained itself in the sludge for at least 45 days and was present in the sludge flocs. After an initial adaptation period of 6 days, complete degradation of 3-CA was obtained during 2 weeks, while no degradation at all occurred in the noninoculated control reactor. Upon further operation of the SCAS system, only 50% 3-CA removal was observed. Denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes revealed a dynamic change in the microbial community structure of the activated sludge. The DGGE patterns of the noninoculated and the inoculated reactors evolved after 7 days to different clusters, which suggests an effect of strain inoculation on the microbial community structure. The results indicate that bioaugmentation, even with a strain originating from that ecosystem and able to effectively grow on a selective substrate, is not permanent and will probably require regular resupplementation.

Identification of prevalent microbial communities in a municipal solid waste landfill

2006 ◽  
Vol 53 (8) ◽  
pp. 139-147 ◽  
Author(s):  
B. Calli ◽  
S. Durmaz ◽  
B. Mertoglu
Keyword(s):  
16S Rrna ◽  
Municipal Solid Waste ◽  
Microbial Communities ◽  
Solid Waste ◽  
Molecular Techniques ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Fish Analysis ◽  
Municipal Solid Waste Landfill ◽  
Waste Landfill

To identify the microbial communities in Istanbul, Odayeri Municipal Solid Waste Landfill, leachate samples were collected from different sections at different stabilization phases. In identification of microbial communities in leachate samples, molecular techniques such as FISH, DGGE and cloning based on 16S rRNA and mcrA genes were used. As the chemical and microbiological compositions of the samples were compared, obvious correlations were found between the stability of the landfill section and abundance of active methanogens. On the other hand, there were considerable differences between acidogenic and mature leachate samples in DGGE profiles of archaeal and bacterial 16S rRNA genes. Moreover, in acidogenic leachate samples having BOD5/COD ratio of about 0.5 acetate utilizing Methanosarcina and Methanosaeta species were intensively detected in FISH. Although only very few H2-utilizing methanogens were identified with FISH analysis, most of the clones isolated from mature leachate samples clustered within H2-utilizing Methanobacteriales and Methanomicrobiales according to phylogenetic analysis of 16S rRNA and mcrA clones, respectively.

scholarly journals Community and Proteomic Analysis of Anaerobic Consortia Converting Tetramethylammonium to Methane

Archaea ◽  
2017 ◽  
Vol 2017 ◽  
pp. 1-14
Author(s):  
Wei-Yu Chen ◽  
Lucia Kraková ◽  
Jer-Horng Wu ◽  
Domenico Pangallo ◽  
Lenka Jeszeová ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
High Throughput Sequencing ◽  
Molecular Techniques ◽  
Upflow Anaerobic Sludge Blanket ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Uasb Reactor ◽  
Anaerobic Sludge ◽  
Methane Formation ◽  
Proteomic Approach

Tetramethylammonium-degrading methanogenic consortia from a complete-mixing suspended sludge (CMSS) and an upflow anaerobic sludge blanket (UASB) reactors were studied using multiple PCR-based molecular techniques and shotgun proteomic approach. The prokaryotic 16S rRNA genes of the consortia were analyzed by quantitative PCR, high-throughput sequencing, and DGGE-cloning methods. The results showed that methanogenicarchaeawere highly predominant in both reactors but differed markedly according to community structure. Community and proteomic analysis revealed thatMethanomethylovoransandMethanosarcinawere the major players for the demethylation of methylated substrates and methane formation through the reduction pathway of methyl-S-CoM and possibly, acetyl-CoA synthase/decarbonylase-related pathways. Unlike high dominance of oneMethanomethylovoranspopulation in the CMSS reactor, diverse methylotrophicMethanosarcinaspecies inhabited in syntrophy-like association with hydrogenotrophicMethanobacteriumin the granular sludge of UASB reactor. The overall findings indicated the reactor-dependent community structures of quaternary amines degradation and provided microbial insight for the improved understanding of engineering application.

scholarly journals Census of the Viral Metagenome within an Activated Sludge Microbial Assemblage

2010 ◽  
Vol 76 (8) ◽  
pp. 2673-2677 ◽  
Author(s):  
Larissa C. Parsley ◽  
Erin J. Consuegra ◽  
Stephen J. Thomas ◽  
Jaysheel Bhavsar ◽  
Andrew M. Land ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
16S Rrna ◽  
Activated Sludge ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Bacterial Isolates ◽  
Microbial Assemblage ◽  
Culture Independent ◽  
Culture Dependent

ABSTRACT The viral metagenome within an activated sludge microbial assemblage was sampled using culture-dependent and culture-independent methods and compared to the diversity of activated sludge bacterial taxa. A total of 70 unique cultured bacterial isolates, 24 cultured bacteriophages, 829 bacterial metagenomic clones of 16S rRNA genes, and 1,161 viral metagenomic clones were subjected to a phylogenetic analysis.

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