A methods comparison for the isolation and detection of thermophilic Campylobacter in agricultural watersheds

被引:20
作者
Khan, Izhar U. H. [1 ]
Gannon, Vic [2 ]
Loughborough, Alyssa [1 ]
Jokinen, Cassandra [2 ]
Kent, Rob [3 ]
Koning, Wendell [4 ]
Lapen, David R. [5 ]
Medeiros, Diane [6 ]
Miller, Jim [7 ]
Neumann, Norman [8 ,9 ]
Phillips, Rob [3 ]
Robertson, Will [6 ]
Schreier, Hans [10 ]
Topp, Edward [11 ]
van Bochove, Eric [12 ]
Edge, Thomas A. [1 ]
机构
[1] Environm Canada, Natl Water Res Inst, Aquat Ecosyst Protect Res Div, Water Sci & Technol Directorate, Burlington, ON L7R 4A6, Canada
[2] Publ Hlth Agcy Canada, Lab Foodborne Zoonoses, Lethbridge, AB, Canada
[3] Environm Canada, Natl Water Qual Monitoring Water Sci & Technol, Gatineau, ON, Canada
[4] Alberta Environm, Calgary, AB, Canada
[5] Agr & Agri Food Canada, Eastern Cereal & Oilseed Res Ctr, Ottawa, ON, Canada
[6] Hlth Canada, Water Air & Climate Change Bur, Ottawa, ON K1A 0L2, Canada
[7] Agr & Agri Food Canada, Lethbridge Res Ctr, Lethbridge, AB, Canada
[8] Univ Alberta, Sch Publ Hlth, Edmonton, AB, Canada
[9] Univ Calgary, Dept Microbiol & Infect Dis, Calgary, AB, Canada
[10] Univ British Columbia, Vancouver, BC V5Z 1M9, Canada
[11] Agr & Agri Food Canada, So Crop Protect & Food Res Ctr, London, ON, Canada
[12] Agr & Agri Food Canada, Soils & Crop Res & Dev Ctr, Quebec City, PQ, Canada
关键词
Thermophilic Campylobacter; Method comparison; Triplex PCR; Agriculture water; PCR ASSAY; JEJUNI; IDENTIFICATION; COLI; VIRULENCE; OUTBREAKS; MEDIA; RIVER; GENE;
D O I
10.1016/j.mimet.2009.09.024
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Campylobacter species contribute to an enormous burden of enteric illnesses around the world. This study compared two different methods for detecting Campylobacter species in surface water samples from agricultural watersheds across Canada. One method was based on membrane filtration (MF) of 500 ml water samples followed by selective microaerophilic enrichment at 42 degrees C in Bolton broth, isolation of Campylobacter on CCDA, and subsequent identification confirmation by a PCR assay. The second method was based on centrifugation (CF) of 1000 ml water samples, followed by selective microaerophilic enrichment at 42 degrees C in Bolton broth, isolation of Campylobacter on Modified Karmali Agar, and subsequent identification confirmation by a different PCR assay. Overall comparison of the CF and MF methods indicated that both methods found Camylobacter jejuni to be the most commonly detected Campylobacter species in 699 water samples from four agricultural watersheds across Canada, and that C. jejuni frequency of occurrence was similar by both methods. However, the CF method detected significantly higher frequencies of Campylobacter coli (17%) and other Campylobacter species (13%) than the MF method (11% and 3%, respectively). It was frequently found that one method would detect Campylobacter in a water sample when the other method would not for a simultaneously collected, duplicate water sample. This study indicates that methods can have significantly different recovery efficiencies for Campylobacter species, and that caution is needed when comparing studies that report on the frequency of occurrence of waterborne Campylobacter at the genus level when different detection methods are used. Crown Copyright (C) 2009 Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:307 / 313
页数:7
相关论文
共 29 条
[1]  
ARVANTIDOU M, 1996, MICROBIOL RES, V50, P153
[2]  
Atabay HI, 1998, J APPL MICROBIOL, V84, P733
[3]   Comparison of three enrichment media for the isolation of Campylobacter spp. from foods [J].
Baylis, CL ;
MacPhee, S ;
Martin, KW ;
Humphrey, TJ ;
Betts, RP .
JOURNAL OF APPLIED MICROBIOLOGY, 2000, 89 (05) :884-891
[4]   A MOST PROBABLE NUMBER METHOD FOR ESTIMATING SMALL NUMBERS OF CAMPYLOBACTERS IN WATER [J].
BOLTON, FJ ;
HINCHLIFFE, PM ;
COATES, D ;
ROBERTSON, L .
JOURNAL OF HYGIENE, 1982, 89 (02) :185-190
[5]  
Clark CG, 2003, EMERG INFECT DIS, V9, P1232
[6]   CULTURE MEDIA FOR THE ISOLATION OF CAMPYLOBACTERS [J].
CORRY, JEL ;
POST, DE ;
COLIN, P ;
LAISNEY, MJ .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 1995, 26 (01) :43-76
[7]   Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli [J].
Denis, M ;
Soumet, C ;
Rivoal, K ;
Ermel, G ;
Blivet, D ;
Salvat, G ;
Colin, P .
LETTERS IN APPLIED MICROBIOLOGY, 1999, 29 (06) :406-410
[8]   The occurrence of campylobacters in water sources in South Africa [J].
Diergaardt, SM ;
Venter, SN ;
Spreeth, A ;
Theron, J ;
Brözel, VS .
WATER RESEARCH, 2004, 38 (10) :2589-2595
[9]   Current epidemiological issues in human campylobacteriosis [J].
Frost, JA .
JOURNAL OF APPLIED MICROBIOLOGY, 2001, 90 :85S-95S
[10]   Specific identification of the enteropathogens Campylobacter jejuni and Campylobacter coli by using a PCR test based on the ceuE gene encoding a putative virulence determinant [J].
Gonzalez, I ;
Grant, KA ;
Richardson, PT ;
Park, SF ;
Collins, MD .
JOURNAL OF CLINICAL MICROBIOLOGY, 1997, 35 (03) :759-763