Cultivation of Campylobacter and Probiotics

Campylobacter jejuni 81-176 is a well-characterized invasive and wild type strain that has been routinely used as a “global model” in studies that characterize C. jejuni virulence and host pathogen interactions (Korlath et al., 1985 (link); Hendrixson and DiRita, 2004 (link); Papp-Szabo et al., 2005 (link); Hofreuter et al., 2006 (link)). In this study, C. jejuni 81-176 was routinely cultured using Mueller-Hinton (MH) agar (Difco) with a Campylobacter selective supplement (CSS) (SR0117; Oxoid) at 42°C under microaerobic conditions (5% O₂, 10% CO₂, and 85% N₂) (Kassem et al., 2012 (link)). E. coli strain Nissle 1917 (EcN) was cultured aerobically using Luria-Bertani (LB) broth at 37°C to achieve logarithmic growth. Other bacterial strains, Lactobacillus rhamnosus GG (LGG; ATCC 53703), Lactobacillus acidophilus NCFM (LA; ATCC 700396), and Bifidobacterium animalis subsp. Lactis (Bb-12; Christian Hansen, Ltd, Hørsholm, Denmark) were cultured using MRS (de Man, Rogosa and Sharpe) media under anaerobic condition, which was generated using the GasPak^TM EZ Anaerobe Container System Sachets (BD, United States) (Kassem et al., 2012 (link)). To facilitate the growth of Bb-12, the MRS broth was supplemented with 0.05% cysteine hydrochloride. LA, LGG, and Bb-12 were grown at 37°C for 18 h (Kumar et al., 2014 (link)).