Nurmi-type Culture Prepared using Culture Media without l-Cysteine Enhances Salmonella Exclusion in Hatched Layer Chicks

To determine the influence of media composition on Salmonella exclusion of Nurmi-type cultures, two and four types of cultures in the first and second trial, respectively, were prepared from the cecal contents of conventional laying hens, and Salmonella exclusion was assessed in newly hatched chicks. In the first trial, modified Viande Levure (VL) broth or nutrient broth (NB) were used to prepare Nurmi-type cultures (N-VL and N-NB), which were administered to the newly hatched chicks. Twenty-four hours later, the chicks were challenged with Salmonella enterica Typhimurium EF85-9 (ST). ST recoveries (log(10) colony forming units/g of cecal contents) from the N-VL-, N-NB-, and control-treated groups 5 days after the challenge were 7.6±0.6, 0.9±1.9, and 7.7±0.4, respectively. The results suggested the influence of l-cysteine (Cys) present in the VL broth. Thus, we determined the effect of Cys in the second trial. We prepared two other cultures using VL broth without Cys (N-VL−Cys) and NB with Cys (NNB−Cys). ST recoveries from the cecal contents of the N-VL-, N-VL−Cys-, and control-treated groups were 6.3±0.9, 2.1±2.5, and 9.2±0.8, respectively. ST was not recovered from the N-NB- and N-NB−Cys-treated groups. To identify bacteria with Salmonella exclusion activity, we isolated 41 bacterial strains from the ceca of N-NB-treated chicks without Salmonella challenge. Most isolates were identified as Enterococcus faecalis or E. mundtii based on 16S rRNA gene sequencing, and only four cultures excluded Salmonella. Therefore, VL broth containing Cys was not always required for preparing Nurmi-type cultures. The use of media prepared with Cys at the lowest possible concentration or without Cys would promote to enhance Salmonella exclusion from Nurmi-type cultures.