Rapid in situ hybridization technique using 16S rRNA segments for detecting and differentiating the closely related gram-positive organisms Bacillus polymyxa and Bacillus macerans

Jurtshuk, R; Blick, Mark; Bresser, Joel; Fox, George E.; Jurtshuk, Peter

doi:10.1128/aem.58.8.2571-2578.1992

Cited by 44 publications

(16 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Many Gram-positive bacteria are dif®cult to permeabilize after ®xation with PFA (Stahl and Amann, 1991;Macnaughton et al, 1994;Amann et al, 1995). Gram-positive bacteria were detected after ®xation by heat (Jurtshuk et al, 1992), ethanol±formalin (9:1 v/v); Braun-Howland et al, 1992) or 50% ethanol (Roller et al, 1994). Treatment of PFA-®xed cells with cell wall lytic enzymes, such as lysozyme or mutanolysin, also increased the permeability of Gram-positive lactococci, enterococci and streptococci (Beimfohr et al, 1993), Streptomyces scabies hyphae (Hahn et al, 1992), and Microthrix parvicella (Erhart et al, 1997).…”

Section: Optimization Of Conditions For¯uorescence In Situ Hybridizatmentioning

confidence: 99%

Desulfotomaculum genus‐ and subgenus‐specific 16S rRNA hybridization probes for environmental studies

Hristova¹,

Mau²,

Zheng³

et al. 2000

Environmental Microbiology

View full text Add to dashboard Cite

Based on comparative analysis of 16S rRNA sequences and the recently established phylogeny of the genus Desulfotomaculum, a set of phylogenetically nested hybridization probes was developed and characterized. A genus-specific probe targets all known Desulfotomaculum species (with the exception of Desulfotomaculum acetoxidans), and five specific probes target subclusters within the Desulfotomaculum genus. The dissociation temperature of each probe was determined experimentally. Probe specificities were verified through hybridizations with pure culture rRNA isolated from a wide variety of target and non-target organisms and through an evaluation of probe 'nesting' using samples obtained from four different environments. Fixation and hybridization conditions for fluorescence in situ hybridizations were also optimized. The probes were used in quantitative membrane hybridizations to determine the abundance of Desulfotomaculum species in thermophilic anaerobic digesters, in soil, in human faeces and in pig colon samples. Desulfotomaculum rRNA accounted for 0.3-2.1% of the total rRNA in the digesters, 2.6-6.6% in soil, 1.5-3.3% in human faeces and 2.5-6.2% in pig colon samples.

show abstract

Section: Optimization Of Conditions For¯uorescence In Situ Hybridizatmentioning

confidence: 99%

Desulfotomaculum genus‐ and subgenus‐specific 16S rRNA hybridization probes for environmental studies

Hristova¹,

Mau²,

Zheng³

et al. 2000

Environmental Microbiology

View full text Add to dashboard Cite

show abstract

“…Since the cell walls of "M. parvicella" and nocardioform actinomycetes are difficult to be penetrated by the oligonucleotide probes, different fixation and permeabilization protocols were tested: Fixation with ethanol [10] or paraformaldehyde (PFA) [6]; additional treatment as combinations of heat treatment (90°C, 30 min) [11] and the application of lysozyme [12] and mutanolysin [5] at different concentrations and incu-…”

Section: Fluorescence In Situ Hybridization (Fish)mentioning

confidence: 99%

Dynamics of Population and Scumming on a Full‐scale Wastewater Treatment Plant in Switzerland

Hug

Ziranke

Siegrist

2005

Acta hydrochim. hydrobiol.

View full text Add to dashboard Cite

SwitzerlandInstitute for Aquatic Science and Technology, Überlandstr.The formation of stable scum (activated sludge foaming) is a serious operational problem 133, Postfach 611, CH-8600 in wastewater treatment plants (WWTP) all over the world. Although scumming seems to Dübendorf, Switzerland be related to high numbers of "Microthrix parvicella" and nocardioform actinomycetes, it is still unknown whether these organisms are necessary and sufficient for the formation of stable scum. To tackle this question, the abundance of these organisms and the scum coverage of the activated sludge basins were measured on a WWTP in Switzerland for almost two years. Using fluorescence in situ hybridization we were able to recognize "M. parvicella" as well as filamentous and non-filamentous types of nocardioform actinomycetes. A newly developed rapid quantification method allowed us to measure seasonal variations of the population. The highly variable scumming followed only partly the abundance of the filamentous types of nocardioform actinomycetes and was not correlated to "M. parvicella". Neither these organisms nor any plant operating condition could fully explain the dynamics of the scumming. The data lead us to suggest that high numbers of "M. parvicella" or nocardioform actinomycetes are not sufficient to cause scumming. The measured dynamics of the population and the scum coverage also imply that, to investigate scumming at a particular WWTP, time-series of measurements including different seasons and periods of different scumming intensities are necessary. Dynamik der Population und der Schaumbildung in einer Schweizer Kläranlage

show abstract

“…This technique has been used extensively in the clinical field as a means of locating specific genes in human chromosomes, and can also be used to locate viral particles within human cells. A number of research groups have exploited ISH to detect individual bacterial ce11s (32,33,34). ISH relies on the use of short, labelled nucleic acid probes to bind to specific gene sequences within cells.…”

Section: In Sztu Hybridization (Ish)mentioning

confidence: 99%

Prospects for New Techniques for Rapid Bacteriological Monitoring of Drinking Water

Sidorowicz

Whitmore

1995

Water & Environment J

View full text Add to dashboard Cite

The microbiological safety of water supplies is at present assured by monitoring for the absence of the faecal indicator organisms, total coliform bacteria and Escherichia coli. This monitoring work represents the bulk of the workload of microbiology laboratories in the water industry. The accuracy of current methods is not questioned, but the times taken to obtain a presumptive result (18 h) and a confirmed result (up to 72 h) impose delays, particularly with regard to the recommissioning of distribution systems. There is therefore a continued demand for more rapid techniques, which has not been satisfied, despite the advances in analytical methods which have been adopted in other industries. Alternatives to the traditional culture‐based methods are reviewed in this paper, and promising techniques for water applications are highlighted. Highly specific bacterial labelling methods and sensitive instrumentation for detection and enumeration already exist, so that rapid same‐day detection of coliforms is technically feasible. Suitable separation methods for rapid analysis of water samples have not yet been developed, but antibody‐mediated separations which are now employed in clinical and food microbiology may be applicable.

show abstract

Rapid in situ hybridization technique using 16S rRNA segments for detecting and differentiating the closely related gram-positive organisms Bacillus polymyxa and Bacillus macerans

Cited by 44 publications

References 14 publications

Desulfotomaculum genus‐ and subgenus‐specific 16S rRNA hybridization probes for environmental studies

Desulfotomaculum genus‐ and subgenus‐specific 16S rRNA hybridization probes for environmental studies

Dynamics of Population and Scumming on a Full‐scale Wastewater Treatment Plant in Switzerland

Prospects for New Techniques for Rapid Bacteriological Monitoring of Drinking Water

Contact Info

Product

Resources

About