Open AccessComparison of Arcobacter Isolation Methods, and Diversity of Arcobacter spp. in Cheshire, United Kingdom
Author(s) -
J. Yvette Merga,
A. J. H. Leatherbarrow,
Craig Winstanley,
Malcolm J. Bennett,
C. A. Hart,
William G. Miller,
Nicola Williams
Publication year - 2011
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.01964-10
Subject(s) - arcobacter , multilocus sequence typing , biology , feces , microbiology and biotechnology , campylobacter , isolation (microbiology) , veterinary medicine , typing , genotype , genetics , bacteria , medicine , gene
The aims of this study were, firstly, to compare five published methods for the isolation ofArcobacter spp. from animal feces in order to determine the most sensitive and specific method. Second, we analyzed the resulting isolates by multilocus sequence typing (MLST) in order to investigate the diversity of the isolates recovered. Third, we investigated the ability to recoverArcobacter spp. from frozen fecal samples. Seventy-seven fecal samples from cattle, sheep, and badgers were subjected to five isolation methods, based on published methods for the isolation ofArcobacter andCampylobacter spp. Thirty-nineArcobacter butzleri isolates were analyzed using a multilocus sequence typing scheme. The survival ofArcobacter spp. in frozen samples was investigated by freezing the fecal samples at −80°C for 7 days and then applying the same five isolation methods. The most sensitive and specific method used anArcobacter -specific broth in conjunction with modified charcoal cefoperazone deoxycholate agar (mCCDA) with added antibiotics. Freezing of fecal samples led to a reduction in the recovery ofArcobacter spp. by approximately 50%. The 39 allelic profiles obtained by MLST could be divided into 11 sequence types (STs). We have identified the most sensitive and specific method for the isolation ofArcobacter spp. from animal feces and demonstrated that the freezing of fecal samples prior to isolation reduces arcobacter recovery. MLST analysis of the isolates revealed a high level of diversity.