scholarly journals Rapid and Sensitive Quantification of Vibrio cholerae and Vibrio mimicus Cells in Water Samples by Use of Catalyzed Reporter Deposition FluorescenceIn SituHybridization Combined with Solid-Phase Cytometry

2012 ◽  
Vol 78 (20) ◽  
pp. 7369-7375 ◽  
Author(s):  
Sonja Schauer ◽  
Regina Sommer ◽  
Andreas H. Farnleitner ◽  
Alexander K. T. Kirschner
Keyword(s):  
Vibrio Cholerae ◽  
Water Samples ◽  
Solid Phase ◽  
Soda Lake ◽  
Environmental Water ◽  
Relative Precision ◽  
Content Type ◽  
Vibrio Mimicus ◽  
Catalyzed Reporter Deposition ◽  
Card Fish

ABSTRACTA new protocol for rapid, specific, and sensitive cell-based quantification ofVibrio cholerae/Vibrio mimicusin water samples was developed. The protocol is based on catalyzed reporter deposition fluorescencein situhybridization (CARD-FISH) in combination with solid-phase cytometry. For pure cultures, we were able to quantify down to 6V. choleraecells on one membrane with a relative precision of 39% and down to 12 cells with a relative precision of 17% after hybridization with the horseradish peroxidase (HRP)-labeled probe Vchomim1276 (specific forV. choleraeandV. mimicus) and signal amplification. The corresponding position of the probe on the 16S rRNA is highly accessible even when labeled with HRP. For the first time, we were also able to successfully quantifyV. cholerae/V. mimicusvia solid-phase cytometry in extremely turbid environmental water samples collected in Austria. Cell numbers ranged from 4.5 × 101cells ml−1in the large saline lake Neusiedler See to 5.6 × 104cells ml−1in an extremely turbid shallow soda lake situated nearby. We therefore suggest CARD-FISH in combination with solid-phase cytometry as a powerful tool to quantifyV. cholerae/V. mimicusin ecological studies as well as for risk assessment and monitoring programs.

scholarly journals RNA Colony Blot Hybridization Method for Enumeration of Culturable Vibrio cholerae and Vibrio mimicus Bacteria

2009 ◽  
Vol 75 (17) ◽  
pp. 5439-5444 ◽  
Author(s):  
Christopher J. Grim ◽  
Young-Gun Zo ◽  
Nur A. Hasan ◽  
Afsar Ali ◽  
Wasimul B. Chowdhury ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Sensitivity And Specificity ◽  
Water Samples ◽  
Blot Hybridization ◽  
False Negative ◽  
Sodium Dodecyl ◽  
Environmental Water ◽  
Health Clinic ◽  
Vibrio Mimicus ◽  
Hybridization Protocol

ABSTRACT A species-specific RNA colony blot hybridization protocol was developed for enumeration of culturable Vibrio cholerae and Vibrio mimicus bacteria in environmental water samples. Bacterial colonies on selective or nonselective plates were lysed by sodium dodecyl sulfate, and the lysates were immobilized on nylon membranes. A fluorescently labeled oligonucleotide probe targeting a phylogenetic signature sequence of 16S rRNA of V. cholerae and V. mimicus was hybridized to rRNA molecules immobilized on the nylon colony lift blots. The protocol produced strong positive signals for all colonies of the 15 diverse V. cholerae-V. mimicus strains tested, indicating 100% sensitivity of the probe for the targeted species. For visible colonies of 10 nontarget species, the specificity of the probe was calculated to be 90% because of a weak positive signal produced by Grimontia (Vibrio) hollisae, a marine bacterium. When both the sensitivity and specificity of the assay were evaluated using lake water samples amended with a bioluminescent V. cholerae strain, no false-negative or false-positive results were found, indicating 100% sensitivity and specificity for culturable bacterial populations in freshwater samples when G. hollisae was not present. When the protocol was applied to laboratory microcosms containing V. cholerae attached to live copepods, copepods were found to carry approximately 10,000 to 50,000 CFU of V. cholerae per copepod. The protocol was also used to analyze pond water samples collected in an area of cholera endemicity in Bangladesh over a 9-month period. Water samples collected from six ponds demonstrated a peak in abundance of total culturable V. cholerae bacteria 1 to 2 months prior to observed increases in pathogenic V. cholerae and in clinical cases recorded by the area health clinic. The method provides a highly specific and sensitive tool for monitoring the dynamics of V. cholerae in the environment. The RNA blot hybridization protocol can also be applied to detection of other gram-negative bacteria for taxon-specific enumeration.

scholarly journals Total and Viable Legionella pneumophila Cells in Hot and Natural Waters as Measured by Immunofluorescence-Based Assays and Solid-Phase Cytometry

2011 ◽  
Vol 77 (17) ◽  
pp. 6225-6232 ◽  
Author(s):  
N. Parthuisot ◽  
M. Binet ◽  
A. Touron-Bodilis ◽  
C. Pougnard ◽  
P. Lebaron ◽  
...  
Keyword(s):  
Legionella Pneumophila ◽  
Water Samples ◽  
Natural Waters ◽  
Solid Phase ◽  
Environmental Water ◽  
Thermal Spring ◽  
Hot Water ◽  
Content Type ◽  
Thermal Spring Water ◽  
Plate Counts

ABSTRACTA new method was developed for the rapid and sensitive detection of viableLegionella pneumophila. The method combines specific immunofluorescence (IF) staining using monoclonal antibodies with a bacterial viability marker (ChemChrome V6 cellular esterase activity marker) by means of solid-phase cytometry (SPC). IF methods were applied to the detection and enumeration of both the total and viableL. pneumophilacells in water samples. The sensitivity of the IF methods coupled to SPC was 34 cells liter−1, and the reproducibility was good, with the coefficient of variation generally falling below 30%. IF methods were applied to the enumeration of total and viableL. pneumophilacells in 46 domestic hot water samples as well as in cooling tower water and natural water samples, such as thermal spring water and freshwater samples. Comparison with standard plate counts showed that (i) the total direct counts were always higher than the plate counts and (ii) the viable counts were higher than or close to the plate counts. With domestic hot waters, when the IF assay was combined with the viability test, SPC detected up to 3.4 × 103viable but nonculturableL. pneumophilacells per liter. These direct IF methods could be a powerful tool for high-frequency monitoring of domestic hot waters or for investigating the occurrence of viableL. pneumophilain both man-made water systems and environmental water samples.

scholarly journals A Novel Triplex Quantitative PCR Strategy for Quantification of Toxigenic and Nontoxigenic Vibrio cholerae in Aquatic Environments

2015 ◽  
Vol 81 (9) ◽  
pp. 3077-3085 ◽  
Author(s):  
Rupert Bliem ◽  
Sonja Schauer ◽  
Helga Plicka ◽  
Adelheid Obwaller ◽  
Regina Sommer ◽  
...  
Keyword(s):  
Risk Assessment ◽  
Vibrio Cholerae ◽  
Quantitative Pcr ◽  
Water Samples ◽  
Dynamic Range ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Aquatic Environments ◽  
Ecological Studies ◽  
Content Type

ABSTRACTVibrio choleraeis a severe human pathogen and a frequent member of aquatic ecosystems. Quantification ofV. choleraein environmental water samples is therefore fundamental for ecological studies and health risk assessment. Beside time-consuming cultivation techniques, quantitative PCR (qPCR) has the potential to provide reliable quantitative data and offers the opportunity to quantify multiple targets simultaneously. A novel triplex qPCR strategy was developed in order to simultaneously quantify toxigenic and nontoxigenicV. choleraein environmental water samples. To obtain quality-controlled PCR results, an internal amplification control was included. The qPCR assay was specific, highly sensitive, and quantitative across the tested 5-log dynamic range down to a method detection limit of 5 copies per reaction. Repeatability and reproducibility were high for all three tested target genes. For environmental application, global DNA recovery (GR) rates were assessed for drinking water, river water, and water from different lakes. GR rates ranged from 1.6% to 76.4% and were dependent on the environmental background. Uncorrected and GR-correctedV. choleraeabundances were determined in two lakes with extremely high turbidity. Uncorrected abundances ranged from 4.6 × 102to 2.3 × 104cell equivalents liter−1, whereas GR-corrected abundances ranged from 4.7 × 103to 1.6 × 106cell equivalents liter−1. GR-corrected qPCR results were in good agreement with an independent cell-based direct detection method but were up to 1.6 log higher than cultivation-based abundances. We recommend the newly developed triplex qPCR strategy as a powerful tool to simultaneously quantify toxigenic and nontoxigenicV. choleraein various aquatic environments for ecological studies as well as for risk assessment programs.

Development of immunoaffinity solid phase microextraction rods for analysis of three estrogens in environmental water samples

2017 ◽  
Vol 1061-1062 ◽  
pp. 41-48 ◽  
Author(s):  
Cuicui Wang ◽  
Linyan Yang ◽  
Na Li ◽  
Xinda Zhang ◽  
Yongze Guo ◽  
...  
Keyword(s):  
Water Samples ◽  
Solid Phase Microextraction ◽  
Solid Phase ◽  
Environmental Water ◽  
Environmental Water Samples

Modern nanobiotechnologies for efficient detection and remediation of mercury

Sensor Review ◽  
2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Mulayam Singh Gaur ◽  
Rajni Yadav ◽  
Mamta Kushwah ◽  
Anna Nikolaevna Berlina
Keyword(s):  
Heavy Metals ◽  
Water Samples ◽  
Low Cost ◽  
High Sensitivity ◽  
Environmental Water ◽  
Mercury Ions ◽  
Content Type ◽  
Efficient Detection ◽  
Sensitivity And Selectivity ◽  
Selection Of

Purpose This information will be useful in the selection of materials and technology for the detection and removal of mercury ions at a low cost and with high sensitivity and selectivity. The purpose of this study is to provide the useful information for selection of materials and technology to detect and remove the mercury ions from water with high sensitivity and selectivity. The purpose of this study is to provide the useful information for selection of materials and technology to detect and remove the mercury ions from water with high sensitivity and selectivity. Design/methodology/approach Different nano- and bio-materials allowed for the development of a variety of biosensors – colorimetric, chemiluminescent, electrochemical, whole-cell and aptasensors – are described. The materials used for their development also make it possible to use them in removing heavy metals, which are toxic contaminants, from environmental water samples. Findings This review focuses on different technologies, tools and materials for mercury (heavy metals) detection and remediation to environmental samples. Originality/value This review gives up-to-date and systemic information on modern nanotechnology methods for heavy metal detection. Different recognition molecules and nanomaterials have been discussed for remediation to water samples. The present review may provide valuable information to researchers regarding novel mercury ions detection sensors and encourage them for further research/development.

scholarly journals Isolation of viable but nonculturable Vibrio cholerae O1 from environmental water samples in Kolkata, India, in a culturable state

2014 ◽  
Vol 3 (2) ◽  
pp. 239-246 ◽  
Author(s):  
Mitsutoshi Senoh ◽  
Jayeeta Ghosh‐Banerjee ◽  
Tamaki Mizuno ◽  
Sumio Shinoda ◽  
Shin‐ichi Miyoshi ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Water Samples ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Viable But Nonculturable ◽  
Vibrio Cholerae O1
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