TABLE 1.
Organisms in the mixture and their growth/recovery media and titers
| Organism (ATCC no.) | Growth/recovery medium or host cell line | Procedure for culture and prepn of stock | Viability titer in stock |
|---|---|---|---|
| Mycobacterium terrae pBEN genetically modified in-house (ATCC 15755) | Middlebrook 7H11 agar, OADC,a and kanamycin (10 μg/ml); incubation 20 days at 36 ± 1°C | 7H9 broth with ADCb and glycerol; cells washed and resuspended in deionized water (8 ml) in a Bijoux bottle (Wheaton, Millville, NJ) with glass beads (Sigma-Aldrich; 3 mm in diam; catalog no. Z143928) and stored at 4°C | 3.7 × 109 CFU/ml |
| Geobacillus stearothermophilus (ATCC 12980) | Trypticase soy agar plates incubated at 56°C for 48 h | Spores heat shocked at 100°C for 45 min, washed in deionized H2O, and stored at 4°C | 1.5 × 108 CFU/ml |
| Acinetobacter baumannii (ATCC 19606) | Trypticase soy agar plates incubated at 36 ± 1°C for 24 h | Inoculated into Trypticase soy broth and incubated for 24 h at 36 ± 1°C, broth centrifuged, and pellet resuspended in deionized H2O and stored at 4°C | 1.2 × 109 CFU/ml |
| Hepatitis A virus (ATCC VR-1402) | FRhK-4 cells (CRL-1688) infected and incubated for 6 days | Cells grown in MEMc with 7% (vol/vol) fetal bovine serum (Fisher; M33-500) and 1% nonessential amino acids (Gibco; 11140) at 36 ± 1°C, monolayers infected and incubated at 36 ± 1°C for 7 days in medium with no antibiotics, flasks frozen and thawed (thrice), cell lysate centrifuged, and supernatant aliquoted for storage at −80°C | 8 × 108 PFU/ml |
a
OADC, oleic acid-albumin-dextrose-catalase.
b
ADC, albumin dextrose-catalase.
c
MEM, minimal essential medium.