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

scholarly journals Variants of ctxB alleles of Vibrio cholerae O1 caused sequential cholera outbreaks in the tribal areas of Odisha, India

2021 ◽  
Author(s):  
Bibhuti Bhusan Pal ◽  
Smruti Ranjan Nayak ◽  
Ashish Kumar Nayak ◽  
Dipti Ranjan Behera ◽  
Swatishree Pany ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Water Samples ◽  
Environmental Water ◽  
High Morbidity ◽  
Rectal Swabs ◽  
Outbreak Areas ◽  
Vibrio Cholerae O1 ◽  
El Tor ◽  
Tribal Areas ◽  
The Relationship

Abstract Cholera localized outbreaks/epidemics accounting for high morbidity and mortality have been reported in different years both from the coastal and tribal districts of Odisha. In the present study, the emergence and spread of two sequential cholera outbreaks reported in July to October 2012 from Rayagada and Kalahandi districts of Odisha was investigated. Environmental water samples from different sources and rectal swabs from diarrhoea patients were analysed for identification, antibiogram profiles and molecular studies using DMAMA-PCR assays. The pulsed field gel electrophoresis (PFGE) was done on some selected Vibrio cholerae O1 strains isolated from these cholera outbreak areas. Results showed 42% of rectal swabs and 2.3% of water samples collected from both the districts were positive for Vibrio cholerae O1 Ogawa biotype El Tor carrying both ctxB1 and ctxB7 genotypes. The common resistance profile of V. cholerae O1 strains was ampicillin, nalidixic acid, furazolidone and co-trimoxazole. The PFGE analysis on selected V. cholerae O1 strains of ctxB1 and ctxB7 genotypes showed three pulsotypes with 96% similarity matrix exhibiting the relationship with their respective water sources. Hence, continuous surveillance is highly essential to monitor the antibiogram profile and changing pattern of ctxB genotypes of V. cholerae O1 in this region.

scholarly journals Particle Diffusometry: An Optical Detection Method for Vibrio cholerae Presence in Environmental Water Samples

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Katherine N. Clayton ◽  
Taylor J. Moehling ◽  
Dong Hoon Lee ◽  
Steven T. Wereley ◽  
Jacqueline C. Linnes ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Water Samples ◽  
Detection Method ◽  
Optical Detection ◽  
Environmental Water ◽  
Environmental Water Samples

scholarly journals Quorum-sensing autoinducers resuscitate dormant Vibrio cholerae in environmental water samples

2013 ◽  
Vol 110 (24) ◽  
pp. 9926-9931 ◽  
Author(s):  
S. M. N. Bari ◽  
M. K. Roky ◽  
M. Mohiuddin ◽  
M. Kamruzzaman ◽  
J. J. Mekalanos ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Vibrio Cholerae ◽  
Water Samples ◽  
Environmental Water ◽  
Environmental Water Samples

scholarly journals Detection of Vibrio cholerae by Real-Time Nucleic Acid Sequence-Based Amplification

2007 ◽  
Vol 73 (5) ◽  
pp. 1457-1466 ◽  
Author(s):  
Else M. Fykse ◽  
Gunnar Skogan ◽  
William Davies ◽  
Jaran Strand Olsen ◽  
Janet M. Blatny
Keyword(s):  
Nucleic Acid ◽  
Vibrio Cholerae ◽  
Real Time ◽  
Water Samples ◽  
Environmental Water ◽  
Nucleic Acid Sequence ◽  
Environmental Water Samples ◽  
Dnase Treatment ◽  
Nasba Assay ◽  
Toxigenic Strains

ABSTRACT A multitarget molecular beacon-based real-time nucleic acid sequence-based amplification (NASBA) assay for the specific detection of Vibrio cholerae has been developed. The genes encoding the cholera toxin (ctxA), the toxin-coregulated pilus (tcpA; colonization factor), the ctxA toxin regulator (toxR), hemolysin (hlyA), and the 60-kDa chaperonin product (groEL) were selected as target sequences for detection. The beacons for the five different genetic targets were evaluated by serial dilution of RNA from V. cholerae cells. RNase treatment of the nucleic acids eliminated all NASBA, whereas DNase treatment had no effect, showing that RNA and not DNA was amplified. The specificity of the assay was investigated by testing several isolates of V. cholerae, other Vibrio species, and Bacillus cereus, Salmonella enterica, and Escherichia coli strains. The toxR, groEL, and hlyA beacons identified all V. cholerae isolates, whereas the ctxA and tcpA beacons identified the O1 toxigenic clinical isolates. The NASBA assay detected V. cholerae at 50 CFU/ml by using the general marker groEL and tcpA that specifically indicates toxigenic strains. A correlation between cell viability and NASBA was demonstrated for the ctxA, toxR, and hlyA targets. RNA isolated from different environmental water samples spiked with V. cholerae was specifically detected by NASBA. These results indicate that NASBA can be used in the rapid detection of V. cholerae from various environmental water samples. This method has a strong potential for detecting toxigenic strains by using the tcpA and ctxA markers. The entire assay including RNA extraction and NASBA was completed within 3 h.

scholarly journals Simultaneous direct detection of toxigenic and non-toxigenic Vibrio cholerae from rectal swabs and environmental samples by sandwich ELISA

2007 ◽  
Vol 56 (10) ◽  
pp. 1340-1345 ◽  
Author(s):  
Urmil Tuteja ◽  
Sanjay Kumar ◽  
Jyoti Shukla ◽  
Joseph Kingston ◽  
Harsh V. Batra
Keyword(s):  
Vibrio Cholerae ◽  
Water Samples ◽  
Direct Detection ◽  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Simultaneous Detection ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Bacterial Strains ◽  
Rectal Swabs

A mAb-based simple, specific and rapid two-tip dipstick ELISA was developed for simultaneous detection of toxin- and non-toxin-producing strains of Vibrio cholerae, and for direct detection of V. cholerae from rectal swabs of patients and from environmental water samples. Rabbit polyclonal antibodies and murine mAbs were raised against recombinant protein (r-protein) antigens of cholera toxin B (CtxB) and outer membrane protein W (OmpW). Rabbit polyclonal antibodies to both r-proteins were coated individually onto the tips of nitrocellulose (NC) membranes of a two-tipped NC dipstick as capture antibodies and a mixture of two mAbs was used for the detecting antibodies. The test was found to be specific for V. cholerae strains O1, O139, non-O1 and non-O139, and did not show any cross-reaction to closely related bacterial strains. The test was evaluated on rectal swabs collected at the bedside of 75 hospitalized diarrhoeal patients and on 50 environmental water samples after enrichment for 4 h in alkaline peptone water. The mAb two-tip dipstick ELISA detected V. cholerae in 52/75 rectal swabs and 2/50 environmental water samples for CtxB antigen, and in 1/50 environmental water samples for the non-toxin OmpW antigen of V. cholerae within 1.5 h. These findings were identical to those observed using PCR and conventional culture methods. Thus, this mAb-based two-tip dipstick ELISA could be used for early and reliable simultaneous detection of toxigenic and non-toxigenic strains of V. cholerae from clinical 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.

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