The Campylobacter jejuni strains shown in S1 and S2 Tables were isolated and characterized by Luangtongkum et al. [22 (link)], and were stored at -80°C in 80% Mueller Hinton broth (MHB: Oxoid, UK) with 20% glycerol. They were then grown on Mueller-Hinton agar (MHA; Oxoid, UK) at 42°C under microaerobic conditions (5% O2, 10% CO2, 85% N2) for 24 h. The second passage from each culture was used in the experiments. When necessary, MHA was supplemented with selective medium (SR01176; Oxoid, UK) and growth medium (SR0232E; Oxoid, UK) (MHA-SS), 30 mg/L kanamycin (Merck, Germany), or 4 mg/L ciprofloxacin (Merck, Germany). The Vibrio harveyi BB170 reporter strain [19 (link),23 (link)] was grown on autoinducer bioassay (AB) medium at 30°C, which contained 17 g/L NaCl (Merck, Germany), 12.3 g/L MgSO4 (Merck, Germany), 2 g/L casamino acids (BD Bacto; Fisher Scientific), 1 mM K2HPO4 (Kemika, Croatia), 0.1 mM L-arginine (Sigma Aldrich, Germany), and 1% (v/v) glycerol (Kemika, Croatia).
Sr0232e
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The SR0232E is a laboratory centrifuge designed for general-purpose applications. It features a maximum speed of 6,000 RPM and a maximum RCF of 3,461 x g. The centrifuge accommodates a variety of rotor types and has a compact, benchtop design.
Automatically generated - may contain errors
Lab products found in correlation
Glycerol, by Kemika (2 mentions)
Mueller hinton broth (mhb), by Thermo Fisher Scientific (2 mentions)
Sr01176, by Thermo Fisher Scientific (2 mentions)
K2hpo4, by Kemika (2 mentions)
Mueller hinton agar (mha), by Thermo Fisher Scientific (2 mentions)
L arginine, by Merck Group (2 mentions)
Sodium chloride (nacl), by Merck Group (2 mentions)
Mgso4, by Merck Group (2 mentions)
Kanamycin, by Merck Group (2 mentions)
Mueller hinton agar (mha), by Merck Group (1 mentions)
Ciprofloxacin, by Merck Group (1 mentions)
Campylobacter growth supplement, by Thermo Fisher Scientific (1 mentions)
Cm0001b, by Thermo Fisher Scientific (1 mentions)
3 protocols using sr0232e
1
Cultivation of Campylobacter and Vibrio strains
2
Cultivation and Storage of Campylobacter and Vibrio Strains
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Campylobacter jejuni NCTC 11168 and C. jejuni 11168ΔluxS [19 (link),27 (link)] provided by Klančnik A. and Bezek K. (University of Ljubljana) were stored at −80 °C in 20% glycerol and 80% Mueller–Hinton broth (MHB; Oxoid, UK). They were grown on Mueller–Hinton agar (MHA; Oxoid, UK) at 42 °C under microaerobic conditions (5% O2, 10% CO2, 85% N2) for 24 h. The second passage from each culture was used in the experiments. When necessary, MHA was supplemented with selective medium (SR01176; Oxoid, UK) and growth medium (SR0232E; Oxoid, UK) (MHA-SS) or 30 mg/L kanamycin (Merck, Darmstadt, Germany). The Vibrio harveyi BB170 reporter strain [27 (link),28 (link)] was grown on autoinducer bioassay (AB) medium at 30 °C, which contained 17 g/L NaCl (Merck, Darmstadt, Germany), 12.3 g/L MgSO4 (Merck, Darmstadt, Germany), 2 g/L casamino acids (BD Bacto, Fisher Scientific, Hampton, NH, USA), 1 mM K2HPO4 (Kemika, Zagreb, Croatia), 0.1 mM L-arginine (Sigma Aldrich, Steinheim am Albuch, Germany), and 1% (v/v) glycerol (Kemika, Zagreb, Croatia).
3
Preparation of Campylobacter Cell Suspensions
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Cultures were prepared from frozen stocks by aseptically placing one bead (TSC, Lancashire, UK) of each isolate in 25 ml Hunts broth containing 0.65 g nutrient broth (CM0001B, Oxoid, Cambridge, UK) and 0.15 g Yeast extract (CM0019B, Oxoid, Cambridge, UK), 5% Lysed Horse Blood (SR048C, Lennox, Dublin), and 0.4% Campylobacter growth supplement (SR0232E, Oxoid, Cambridge, UK). The inoculated broths were incubated under microaerobic conditions at 42°C for 48 h. After incubation, broths were vortexed for 30 s followed by centrifugation at 10,000 rpm for 10 min at 4°C. The supernatant was discarded and the pellet was suspended in 25 ml phosphate-buffered saline (P4417, Sigma-Aldrich Arklow, Wicklow, Ireland) and vortexed. Cell suspensions were diluted to 10−3 in 9 ml Maximum Recovery Diluent (MRD, CM0733B Oxoid, Cambridge, UK). A 1 ml volume of the suspensions were then transferred to 99 ml Hunts broth to give a final cell concentration of 3 log10 CFU/ml for spiking of the caeca. Plate counts were carried to confirm spiking concentrations.
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