Application of duplex-PCR in rapid and reliable detection of toxigenic Vibrio cholerae in water samples in Thailand

This study was conducted to assess the bacteriological quality of alternative drinking water supply options in southwest coastal areas of Bangladesh. A total of 90 water samples were collected during both dry and wet seasons from household based rainwater harvesting systems (RWHSs), community based rain water harvesting systems (CRWHSs), pond-sand filters (PSFs) and ponds. The samples were evaluated for faecal coliform, Escherichia coli and Heterotrophic Plate Count, as well as Vibrio cholerae, Salmonella spp., Shigella spp. and Pseudomonas spp. Physico-chemical parameters (pH, electrical conductivity, and color) were also examined. In addition, sanitary inspections were conducted to identify faecal contamination sources. All options showed varying degrees of indicator bacterial contamination. The median E. coli concentrations measured for RWHSs, CRWHSs, PSFs, and ponds were 16, 7, 11, and 488 cfu/100 ml during the wet season, respectively. Vibrio cholerae O1/O139, Salmonella and Shigella spp. were not found in any samples. However, Vibrio cholerae Non-O1/Non-O139 and Pseudomonas spp. were isolated from 74.4% and 91.1% of the water samples collected during the wet season. A maximum pH of 10.4 was found in CRWHSs. Estimation of the disease burden for all options in disability adjusted life years (DALYs) showed an increased disease burden during the wet season. According to sanitary inspections, poor maintenance and unprotected ponds were responsible for rainwater and PSF water contamination, respectively. The findings of the present study suggest that alternative drinking water supply options available in southwest coastal Bangladesh pose a substantial risk to public health.

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Outbreak investigation of Cholera in Shantinagar VDC of Ilam District in Eastern Nepal

International Journal of Infection and Microbiology ◽

10.3126/ijim.v2i3.8666 ◽

2013 ◽

Vol 2 (3) ◽

pp. 87-90

Author(s):

RB Rayamajhi ◽

G Pokharel ◽

G Sharma ◽

B Neupane ◽

VK Khanal ◽

...

Keyword(s):

Vibrio Cholerae ◽

Causative Agent ◽

Water Samples ◽

Group Discussion ◽

Control Measures ◽

Water Tank ◽

District Health ◽

Source Of Infection ◽

Local Residents ◽

Stool Analysis

INTRODUCTION: In 2012 August, suspected cases of Cholera in increased number were reported to the district health office of Ilam by the in charge of Shantinagar health post asking for intervention to stop further occurrences. The aim of the investigation was to identify the possible source of infection, the causative agent and the application of local control measures for prevention of further recurrences. MATERIALS AND METHODS: Focus group discussion and Key informant survey were conducted to assess the sanitary hygiene and practices along with source of drinking water for probable cause of diarrhea. Similarly, face to face interview was done among the sick and healthy local residents to collect the demographic and clinical details. Ten stool and water samples were sent for analysis in Biratnagar, Dharan. RESULTS: 150 local residents had suffered from diarrhea with no deaths. People of all the age group were affected with children being more sufferers. Six of the stool analysis and eight of the water samples were positive for Vibrio Cholerae. It was also observed that sanitary hygiene and practices wasn’t adequate after nose and bottoms cleaning of the children and before preparing the meal. CONCLUSIONS: Lack of adequate cleanliness of the common water tank was found to be the culprit behind the incident and Vibrio Cholerae being the causative agent. Local residents and water management committee were demonstrated the technique of chlorination of water and proper hand washing practices to prevent further recurrences. DOI: http://dx.doi.org/10.3126/ijim.v2i3.8666 Int J Infect Microbiol 2013;2(3):87-90

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Application of a paper based device containing a new culture medium to detect Vibrio cholerae in water samples collected in Haiti

Journal of Microbiological Methods ◽

10.1016/j.mimet.2016.12.014 ◽

2017 ◽

Vol 133 ◽

pp. 23-31 ◽

Cited By ~ 6

Author(s):

Romain Briquaire ◽

Rita R. Colwell ◽

Jacques Boncy ◽

Emmanuel Rossignol ◽

Aline Dardy ◽

...

Keyword(s):

Vibrio Cholerae ◽

Water Samples ◽

Culture Medium

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Semi-nested polymerase chain reaction for detection of toxigenic Vibrio cholerae from environmental water samples

Indian Journal of Microbiology ◽

10.1007/s12088-007-0041-7 ◽

2007 ◽

Vol 47 (3) ◽

pp. 207-211 ◽

Cited By ~ 3

Author(s):

Ajay Kumar Goel ◽

Shweta Bhadauria ◽

Pramod Kumar ◽

Dev V. Kamboj ◽

Lokendra Singh

Keyword(s):

Polymerase Chain Reaction ◽

Vibrio Cholerae ◽

Water Samples ◽

Environmental Water ◽

Environmental Water Samples ◽

Chain Reaction ◽

Nested Polymerase Chain Reaction ◽

Polymerase Chain

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Detection of Vibrio cholerae and Vibrio vulnificus by duplex PCR specific to the groEL gene

Fisheries Science ◽

10.1007/s12562-012-0586-1 ◽

2012 ◽

Vol 79 (2) ◽

pp. 335-340 ◽

Cited By ~ 2

Author(s):

Muhammad Tofazzal Hossain ◽

Yu-Ri Kim ◽

Eun-Young Kim ◽

Jong-Min Lee ◽

In-Soo Kong

Keyword(s):

Vibrio Cholerae ◽

Vibrio Vulnificus ◽

Duplex Pcr ◽

Groel Gene

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Detection of Vibrio cholerae O1 and O139 in Environmental Water Samples by an Immunofluorescent-Aggregation Assay

Applied and Environmental Microbiology ◽

10.1128/aem.02559-09 ◽

2010 ◽

Vol 76 (16) ◽

pp. 5520-5525 ◽

Cited By ~ 15

Author(s):

Duochun Wang ◽

Xuebin Xu ◽

Xiaoling Deng ◽

Changyi Chen ◽

Baisheng Li ◽

...

Keyword(s):

Vibrio Cholerae ◽

Real Time ◽

Water Samples ◽

Real Time Pcr ◽

High Specificity ◽

Culture Method ◽

Environmental Water ◽

Estuarine Water ◽

Conventional Culture ◽

Conventional Culture Method

ABSTRACT Environmental waters are an important reservoir for Vibrio cholerae, and effective surveillance of the pathogen can help to warn of and prevent infection with this potentially fatal pathogen. An immunofluorescent-aggregation (IFAG) assay to detect V. cholerae O1 and O139 was established and evaluated with estuarine water samples. The practical application of this assay was compared with the conventional culture method and real-time PCR. The IFAG method had a sensitivity of 103 CFU/ml for detection of V. cholerae O1 and O139 strains in a suspension containing 10 different species of enterobacterial strains (total, 105 CFU/ml). Ten fluorescent bacterial aggregate colonies were randomly picked and tested positive in serum agglutination tests for the V. cholerae O1 and O139 strains, showing a high specificity. The enrichment broths of 146 samples of estuarine water were tested, and the percentage positive by the IFAG assay was 19.9% (29/146), which was significantly higher than that of the conventional culture method (10.3%, 15/146; P < 0.01) but lower than that of real-time PCR (29.5%, 43/146; P < 0.01). The coincidence rates of real-time PCR and IFAG detection were decreased with the reduction of the V. cholerae concentration. The IFAG method, with a high specificity and a relatively high sensitivity, may be used for detection and isolation of V. cholerae in environmental water samples.

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

Journal of Water and Health ◽

10.2166/wh.2021.126 ◽

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.

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

Scientific Reports ◽

10.1038/s41598-018-38056-7 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 6

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

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

Applied and Environmental Microbiology ◽

10.1128/aem.02007-08 ◽

2009 ◽

Vol 75 (17) ◽

pp. 5439-5444 ◽

Cited By ~ 5

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.

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