scholarly journals Detection of Indigenous HalobacillusPopulations in Damaged Ancient Wall Paintings and Building Materials: Molecular Monitoring and Cultivation

2001 ◽  
Vol 67 (10) ◽  
pp. 4891-4895 ◽  
Author(s):  
Guadalupe Piñar ◽  
Cayo Ramos ◽  
Sabine Rölleke ◽  
Claudia Schabereiter-Gurtner ◽  
Dietmar Vybiral ◽  
...  
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Building Materials ◽  
Dna Analysis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Wall Paintings ◽  
Molecular Monitoring ◽  
Dna Reassociation ◽  
Gradient Gel Electrophoresis ◽  
Spore Forming Bacteria

ABSTRACT Several moderately halophilic gram-positive, spore-forming bacteria have been isolated by conventional enrichment cultures from damaged medieval wall paintings and building materials. Enrichment and isolation were monitored by denaturing gradient gel electrophoresis and fluorescent in situ hybridization. 16S ribosomal DNA analysis showed that the bacteria are most closely related to Halobacillus litoralis. DNA-DNA reassociation experiments identified the isolates as a population of hitherto unknownHalobacillus species.

Analysis of bacterial community in membrane-separation bioreactors by fluorescent in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE) techniques

2000 ◽  
Vol 41 (10-11) ◽  
pp. 259-268 ◽  
Author(s):  
B.S. Luxmy ◽  
F. Nakajima ◽  
Kazuo Yamamoto
Keyword(s):  
In Situ Hybridization ◽  
Bacterial Communities ◽  
Gel Electrophoresis ◽  
Fluorescent In Situ Hybridization ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Membrane Separation ◽  
Diversity Index ◽  
Ammonia Oxidizing Bacteria ◽  
Gradient Gel Electrophoresis

The bacterial communities of membrane-separation bioreactors (MBR) fed with raw sewage were analyzed by a pilot scale study. The community was analyzed by both Fluorescent in Situ Hybridization (FISH) and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) techniques. Five rRNA targeted group specific oligonucleotide probes showed that the alpha- and beta- subclasses of proteobacteria were the most dominant groups among them. The identification of ammonia-oxidizing bacteria in MBR was confirmed by three probes: NEU, Nsv 443 and Nso 190. Mostly the ammonia-oxidizers were found in groups and present in the form of clusters or aggregates. The ratio of NEU/EUB was estimated by double hybridization and image analysis techniques as 6%. The Nitrobacter sp. was also identified inside the MBR with the help of a NIT3 probe and they were also found to be present in the form of a cluster. Usually the clusters formed by the Nitrobacter sp. were smaller than those of ammonia-oxidizing groups. After numerical analysis on the band pattern of DGGE, it was found that the MBR bacterial communities were different from that of conventional activated sludge (CAS) communities with dissimilarity indexes more than 0.6. The diversity of the microbial community was estimated by the Shannon-Weaver index of general diversity. It was found that the value of the diversity index for the CAS process was 1.61 while those for two MBR processes were 1.68 and 1.59.

scholarly journals Novel Epibiotic Thiothrix Bacterium on a Marine Amphipod

2004 ◽  
Vol 70 (6) ◽  
pp. 3772-3775 ◽  
Author(s):  
David C. Gillan ◽  
Nicole Dubilier
Keyword(s):  
Gel Electrophoresis ◽  
Fluorescent In Situ Hybridization ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Rrna Gene ◽  
Filamentous Bacteria ◽  
Gradient Gel Electrophoresis ◽  
Marine Amphipod ◽  
Amphipod Crustacean ◽  
The 16S Rrna Gene

ABSTRACT Comparative analysis of the 16S rRNA gene and fluorescent in situ hybridization (FISH) was used to identify epibiotic filamentous bacteria living on the marine amphipod crustacean Urothoe poseidonis. The epibionts belong to the gamma proteobacteria and represent a novel marine phylotype within the genus Thiothrix. FISH and denaturing gradient gel electrophoresis revealed that the Thiothrix filaments are present on the majority of the amphipods examined.

scholarly journals An Intranuclear Sodalis-Like Symbiont and Spiroplasma Coinfect the Carrot Psyllid, Bactericera trigonica (Hemiptera, Psylloidea)

2020 ◽  
Vol 8 (5) ◽  
pp. 692 ◽  
Author(s):  
Saptarshi Ghosh ◽  
Noa Sela ◽  
Svetlana Kontsedalov ◽  
Galina Lebedev ◽  
Lee R. Haines ◽  
...  
Keyword(s):  
16S Rdna ◽  
Fluorescent In Situ Hybridization ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Tsetse Flies ◽  
Stink Bugs ◽  
Pcr Rflp ◽  
Gradient Gel Electrophoresis ◽  
Candidatus Liberibacter ◽  
Bacterial Plant Pathogen

Endosymbionts harbored inside insects play critical roles in the biology of their insect host and can influence the transmission of pathogens by insect vectors. Bactericera trigonica infests umbelliferous plants and transmits the bacterial plant pathogen Candidatus Liberibacter solanacearum (CLso), causing carrot yellows disease. To characterize the bacterial diversity of B. trigonica, as a first step, we used PCR-restriction fragment length polymorphism (PCR-RFLP) and denaturing gradient gel electrophoresis (DGGE) analyses of 16S rDNA to identify Sodalis and Spiroplasma endosymbionts. The prevalence of both symbionts in field-collected psyllid populations was determined: Sodalis was detected in 100% of field populations, while Spiroplasma was present in 82.5% of individuals. Phylogenetic analysis using 16S rDNA revealed that Sodalis infecting B. trigonica was more closely related to symbionts infecting weevils, stink bugs and tsetse flies than to those from psyllid species. Using fluorescent in situ hybridization and immunostaining, Sodalis was found to be localized inside the nuclei of the midgut cells and bacteriocytes. Spiroplasma was restricted to the cytoplasm of the midgut cells. We further show that a recently reported Bactericera trigonica densovirus (BtDNV), a densovirus infecting B. trigonica was detected in 100% of psyllids and has reduced titers inside CLso-infected psyllids by more than two-fold compared to CLso uninfected psyllids. The findings of this study will help to increase our understanding of psyllid–endosymbiont interactions.

Profiles ofMycobacteriumcommunities under polycyclic aromatic hydrocarbon contamination stress in the Shenfu Irrigation Area, northeast China

2013 ◽  
Vol 59 (10) ◽  
pp. 694-700 ◽  
Author(s):  
Xinyu Li ◽  
Xu Li ◽  
Jian Wang ◽  
Xiujuan Wang ◽  
Jian Sun ◽  
...  
Keyword(s):  
Rice Field ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Community Diversity ◽  
Hydrocarbon Contamination ◽  
Contaminated Sites ◽  
Field Soils ◽  
Polycyclic Aromatic Hydrocarbon Contamination ◽  
Polycyclic Aromatic ◽  
Gradient Gel Electrophoresis

Indigenous Mycobacterium communities play an important role in the degradation of polycyclic aromatic hydrocarbons (PAHs), but little is known about Mycobacterium distribution in situ at PAH-contaminated sites. In this study, the diversity and distribution of Mycobacterium communities were investigated in sediments and soils at sites upstream, midstream, and downstream of an oil-sewage irrigation channel, using denaturing gradient gel electrophoresis (DGGE). The results show that heavy PAH contamination in upstream sites negatively affected Mycobacterium community diversity compared with midstream and downstream sites in all 3 sample types (sediments, corn field soils, and rice field soils). There was a correlation between the distribution of Mycobacterium communities and PAH contamination, as indicated by canonical correspondence analysis. Mycobacterium diversity and distribution was found to vary between the 3 sample types.

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.

Degradation of Nonylphenol Polyethoxylates and its Microflora Structure in an Anoxic-Oxic Activated Sludge Process

2012 ◽  
Vol 610-613 ◽  
pp. 331-336
Author(s):  
Yuan Hua Xie ◽  
Tong Zhu ◽  
Xiao Jiang Liu ◽  
Hui Liu ◽  
Jin Han
Keyword(s):  
In Situ Hybridization ◽  
Activated Sludge ◽  
Removal Efficiency ◽  
Gel Electrophoresis ◽  
Loading Rate ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Oxygen Demand ◽  
Activated Sludge Process ◽  
Gradient Gel Electrophoresis

An anoxic-oxic activated sludge process (AOASP) was carried out to degrade nonylphenol polyethoxylates (NPEOs). The carbon source in influent was replaced stepwise by a mixture of nonylphenol decaethoxylate (M-NP10EO). The 2nd-derivative UV-spectrometry was applied to determine the total amount of M-NP10EO in water samples. Chemical oxygen demand (COD) removal efficiency achieves about 85% under the highest M-NP10EO loading rate, and M-NP10EO removal efficiency is about 80%. Denaturing gradient gel electrophoresis (DGGE) results of activated sludges show that the microbe species decrease but gradually stabilize with the increase of M-NP10EO concentration in influent. Fluorescence in situ hybridization (FISH) results of activated sludges showe that the dominant microflora under the highest M-NP10EO loading rate is β-Proteobacteria (35%), followed by α-Proteobacteria (15%), γ-Proteobacteria (5%) and Actinobateria (4%).

scholarly journals Molecular Monitoring of Succession of Bacterial Communities in Human Neonates

2002 ◽  
Vol 68 (1) ◽  
pp. 219-226 ◽  
Author(s):  
Christine F. Favier ◽  
Elaine E. Vaughan ◽  
Willem M. De Vos ◽  
Antoon D. L. Akkermans
Keyword(s):  
Sequence Analysis ◽  
16S Rdna ◽  
Bacterial Communities ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Bacterial Colonization ◽  
Molecular Monitoring ◽  
Fecal Samples ◽  
16S Rdna Sequence Analysis ◽  
Rdna Sequences ◽  
Gradient Gel Electrophoresis

ABSTRACT The establishment of bacterial communities in two healthy babies was examined for more than the first 10 months of life by monitoring 16S ribosomal DNA (rDNA) diversity in fecal samples by PCR and denaturing gradient gel electrophoresis (DGGE) and by analyzing the sequences of the major ribotypes. DGGE profiles of the dominant populations in the intestines of the infants were obtained by analyzing daily or weekly fecal samples. After delivery, the germfree infant gastrointestinal tracts were rapidly colonized, and the succession of bacteria in each ecosystem was monitored. During the first few days of life the profiles were simple, but they became more complex as the bacterial diversity increased with time in both babies. Clone libraries of amplified 16S rDNA fragments from baby feces were constructed, and these libraries allowed identification of the bacterial types by comparative DNA sequence analysis; the bacteria identified included members of the genera Bifidobacterium, Ruminococcus, Enterococcus, Clostridium, and Enterobacter. Species most closely related to the genera Bifidobacterium and Ruminococcus in particular dominated the intestinal microbiota based on the stability over time and the numbers, as estimated by the intensities of the bands. However, 19 of the 34 cloned rDNA sequences exhibited less than 97% identity with sequences of known bacteria or cloned sequences in databases. This study showed that using PCR-DGGE and 16S rDNA sequence analysis together resulted in a dynamic description of bacterial colonization in the infant intestinal ecosystem and allowed visualization of bacteria that are difficult to cultivate or to detect by other methods.

scholarly journals Characterization of a Culturable Alphaproteobacterial Symbiont Common to Many Marine Sponges and Evidence for Vertical Transmission via Sponge Larvae

2006 ◽  
Vol 72 (5) ◽  
pp. 3724-3732 ◽  
Author(s):  
Julie J. Enticknap ◽  
Michelle Kelly ◽  
Olivier Peraud ◽  
Russell T. Hill
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Marine Sponges ◽  
Rrna Gene ◽  
Surrounding Water ◽  
Axinella Corrugata ◽  
Gradient Gel Electrophoresis ◽  
Related Group

ABSTRACT A closely related group of alphaproteobacteria were found to be present in seven genera of marine sponges from several locations and were shown to be transferred between sponge generations through the larvae in one of these sponges. Isolates of the alphaproteobacterium were cultured from the sponges Axinella corrugata, Mycale laxissima, Monanchora unguifera, and Niphates digitalis from Key Largo, Florida; Didiscus oxeata and Monanchora unguifera from Discovery Bay, Jamaica; an Acanthostronglyophora sp. from Manado, Indonesia; and Microciona prolifera from the Cheasapeake Bay in Maryland. Isolates were very similar to each other on the basis of 16S rRNA gene sequence (>99% identity) and are closely related to Pseudovibrio denitrificans. The bacterium was never isolated from surrounding water samples and was cultured from larvae of M. laxissima, indicating that it is a vertically transmitted symbiont in this sponge. Denaturing gradient gel electrophoresis, 16S rRNA gene clone library analysis, and fluorescent in situ hybridization with probes specific to the alphaproteobacterium confirmed the presence of this bacterium in the M. laxissima larvae. The alphaproteobacterium was densely associated with the larvae rather than being evenly distributed throughout the mesohyl. This is the first report of the successful culture of a bacterial symbiont of a sponge that is transferred through the gametes.

Sign in / Sign up

Export Citation Format

Share Document