title: Characteristics and comparative performance of direct culture, direct PCR and enumeration methods for detection and quantification of Campylobacter spp. in broiler caeca.
abstract: Detection and enumeration of Campylobacter spp. in broiler chicken flocks are key components of research and surveillance studies aimed at reducing Campylobacter infections in people. Direct culture of caecal contents onto selective agar is the typical method used to confirm flock colonisation. Modified charcoal cefoperazone deoxycholate agar (mCCDA) is commonly used for this method, although alternative selective media have been used. Additionally, PCR methods to detect Campylobacter DNA from caecal contents may provide a rapid alternative. However comparative performance data for these methods is limited and therefore required to ensure optimal detection methods for this sample type. In this study, 306 broiler caeca were tested for Campylobacter using direct culture on mCCDA, Skirrows and Preston agars and two real-time PCR methods, one specific for mapA/ceuE regions and another for the flaA gene region. Additionally, the suitability of spread plating and spiral plating methods for enumeration of Campylobacter and the impact of sample storage were assessed. This study confirmed modified CCDA as an optimal media for detection of Campylobacter in broiler caeca. It was significantly more sensitive than Skirrows or Preston agars. This study also demonstrated that the mapA/ceuE PCR had excellent agreement with culture on mCCDA and is a genuine alternative method. Spread plating and spiral plating methods were suitable for enumeration although spiral plating appeared more sensitive for stored samples (72 h). A 1 log reduction in viable Campylobacters was observed in stored samples, therefore storage effects should be considered for quantitative studies with broiler caeca.
authors.affiliation: Department of Bacteriology, Animal Health and Veterinary Laboratories Agency Weybridge, United Kingdom. john.rodgers@ahvla.gsi.gov.uk
authors.forename: J D
authors.initials: JD
authors.lastname: Rodgers
authors.forename: J R
authors.initials: JR
authors.lastname: Lawes
authors.forename: A B
authors.initials: AB
authors.lastname: Vidal
authors.forename: J
authors.initials: J
authors.lastname: Ellis-Iversen
authors.forename: A
authors.initials: A
authors.lastname: Ridley
authors.forename: E J
authors.initials: EJ
authors.lastname: Pleydell
authors.forename: L F
authors.initials: LF
authors.lastname: Powell
authors.forename: M
authors.initials: M
authors.lastname: Toszeghy
authors.forename: K
authors.initials: K
authors.lastname: Stapleton
authors.forename: F A
authors.initials: FA
authors.lastname: Clifton-Hadley
mesh_terms: D000362:Agar; D000818:Animals; D058491:Bacterial Load / Q000379:methods*; D002167:Campylobacter / Q000254:growth & development / Q000302:isolation & purification* / Q000502:physiology; D002169:Campylobacter Infections / Q000662:veterinary*; D002432:Cecum / Q000382:microbiology; D002645:Chickens; D016133:Polymerase Chain Reaction / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D000362:Agar