title: Use of blood-free enrichment broth in the development of a rapid protocol to detect Campylobacter in twenty-five grams of chicken meat.
abstract: A Food Pathogen Enrichment (FPE) broth, which supports the growth of Campylobacter without lysed blood and CO2, was developed. The FPE broth supports the growth of Campylobacter to the same degree as Bolton and Preston broths. Using the FPE broth, we developed a novel rapid protocol to detect small numbers of Campylobacter in 25g of food. The sensitivity of FPE enrichment and PCR to detect Campylobacter spp. from spiked chicken meat was determined. The detection sensitivities for non-stressed C. jejuni and C. coli from fresh meat ranged from 5.8 to 1.1×10(1)CFU per 25g of chicken meat, and those for freeze-stressed C. jejuni and C. coli from frozen meat ranged from 9.9×10(1) to 2.0×10(2)CFU. The FPE broth enrichment culture (24h) of chicken meat, followed by PCR, resulted in a significantly higher detection score (80% positive) than conventional Bolton enrichment and subsequent colony isolation using mCCDA agar plates (18% positive). Differences between our new protocol and the Bolton enrichment method were due to the overgrowth of many resistant bacteria, especially extended-spectrum beta-lactamase-producing bacteria in the Bolton enrichment broth.
authors.affiliation: Department of Microbiology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194, Japan. gif154@gifu-u.ac.jp
authors.forename: Masahiro
authors.initials: M
authors.lastname: Hayashi
authors.forename: Sayoko
authors.initials: S
authors.lastname: Kubota-Hayashi
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Natori
authors.forename: Takuya
authors.initials: T
authors.lastname: Mizuno
authors.forename: Machiko
authors.initials: M
authors.lastname: Miyata
authors.forename: Shigeru
authors.initials: S
authors.lastname: Yoshida
authors.forename: Jiwei
authors.initials: J
authors.lastname: Zhang
authors.forename: Keiko
authors.initials: K
authors.lastname: Kawamoto
authors.forename: Kiyofumi
authors.initials: K
authors.lastname: Ohkusu
authors.forename: Souichi
authors.initials: S
authors.lastname: Makino
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ezaki
mesh_terms: D000818:Animals; D002167:Campylobacter / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002645:Chickens; D016895:Culture Media, Serum-Free; D005516:Food Microbiology / Q000379:methods*; D005615:Freezing; D008460:Meat / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D016895:Culture Media, Serum-Free