INTRODUCTION Campylobacteriosis is a leading global public health issue and is the third most common foodborne disease in the European Union (EFSA, 2013). After a significant decline, the number of human Campylobacter infections in the United States increased in 2012, when the inci- dence was 14.30 cases per 100,000 inhabitants (CDC, 2013). Campylobacter jejuni currently accounts for 90% of the cases of foodborne campylobacteriosis confirmed in the laboratory, followed by C. coli, which causes 8% of the cases (CDC, 2013). Campylobacter exposure in- cludes contact with infected animals and consumption of either untreated water or contaminated foods of ani- mal origin, such as raw milk and meat (EFSA, 2013). Because of the optimal growth temperature and colo- nization ability, thermophilic Campylobacter species have effectively adapted to the avian gut, where they are commonly found in high numbers (Lee and Newell, 2006; Hermans et al., 2012). Although Campylobacter colonization of the broiler gut is not related to clinical disease, it might subsequently lead to contamination of broiler meat during processing (Rosenquist et al., 2006; Hermans et al., 2012). Carcasses generally become contaminated during defeathering or by the leakage of feces from colonized broilers due to eventual gut rup- ture during evisceration (Franchin et al., 2005; Reich et al., 2008). Several studies have reported the high mean prevalence of thermophilic Campylobacter in broiler carcasses (Rosenquist et al., 2006; Kuana et al., 2008; EFSA, 2013). Therefore, handling or consumption of undercooked chicken is considered a major source of human campylobacteriosis (EFSA, 2013). Campylobacter control in broilers is a multifactorial process and involves all steps of the food chain includ- ing primary production (Rosenquist et al., 2006; Reich et al., 2008; Hermans et al., 2012). Therefore, it is im- portant to establish surveillance programs in broiler Isolation of Campylobacter from Brazilian broiler flocks using different culturing procedures C. S. L. Vaz,* 1 D. Voss-Rech,* J. S. Pozza,† A. Coldebella,* and V. S. Silva* *Empresa Brasileira de Pesquisa Agropecuária–Embrapa Suínos e Aves, BR 153, Km 110, 89700-000, Concordia, SC, Brazil; and †Fundação Universidade do Contestado, 89700-000, Concordia, SC, Brazil ABSTRACT Conventional culturing methods enable the detection of Campylobacter in broiler flocks. How- ever, laboratory culture of Campylobacter is laborious because of its fastidious behavior and the presence of competing nontarget bacteria. This study evaluated different protocols to isolate Campylobacter from broil- er litter, feces, and cloacal and drag swabs. Samples taken from commercial Brazilian broiler flocks were di- rectly streaked onto Preston agar (PA), Campy-Line agar (CLA), and modified charcoal cefoperazone deoxy- cholate agar (mCCDA) and also enriched in blood-free Bolton broth (bfBB) for 24 and 48 h followed by plating onto the different selective media. Higher numbers of Campylobacter-positive cloacal and drag swab samples were observed using either direct plating or enrichment for 24 h before plating onto PA, compared with enrich- ment for 48 h (P < 0.05). Furthermore, direct plating was a more sensitive method to detect Campylobacter in broiler litter and feces samples. Analysis of directly plated samples revealed that higher Campylobacter lev- els were detected in feces streaked onto PA (88.8%), cloacal swabs plated onto mCCDA (72.2%), drag swabs streaked onto CLA or mCCDA (69.4%), and litter sam- ples inoculated onto PA (63.8%). Preston agar was the best agar to isolate Campylobacter from directly plated litter samples (P < 0.05), but there was no difference in the efficacies of PA, mCCDA, and CLA in detecting Campylobacter in other samples. The isolated Campylo- bacter strains were phenotypically identified as Campy- lobacter jejuni or Campylobacter coli. The predominant contaminant observed in the Campylobacter cultures was Proteus mirabilis, which was resistant to the major- ity of antimicrobial agents in selective media. Together, these data showed that direct plating onto PA and onto either CLA or mCCDA as the second selective agar enabled the reliable isolation of thermophilic Campylo- bacter species from broiler samples. Finally, Campylo- bacter was detected in all broiler flocks sampled. Key words: Campylobacter jejuni, Campylobacter coli, selective culture, Proteus mirabilis, food safety 2014 Poultry Science 93:2887–2892 http://dx.doi.org/10.3382/ps.2014-03943 PROCESSING, PRODUCTS, AND FOOD SAFETY 2887 Received January 30, 2014. Accepted July 9, 2014. 1 Corresponding author: clarissa.vaz@embrapa.br ©2014 Poultry Science Association Inc. Downloaded from https://academic.oup.com/ps/article-abstract/93/11/2887/1574980 by guest on 30 July 2018