TABLE 1.
List of 53 culture methods including media, additional supplements, and oxygen condition used for swine gut microbiome culturea
| Medium full name | Abbreviation(s) | Additional supplement(s) | Aerobic | Anaerobic |
|---|---|---|---|---|
| Brain heart infusion agar | BHI1 | No supplement | No. 1 | No. 2 |
| BHI3 | 0.5 g/liter l-cysteine hydrochloride hydrate, 10 mg/liter hemin, and 1 mg/liter vitamin K | No. 3 | No. 4 | |
| BHI2 | 10 mg/liter colistin sulfate and 5 mg/liter nalidixic acid | No. 5 | No. 6 | |
| BHI4 | 0.5 g/liter l-cysteine hydrochloride hydrate, 10 mg/liter hemin, 1 mg/liter vitamin K, 10 mg/liter colistin sulfate, and 5 mg/liter nalidixic acid | No. 7 | No. 8 | |
| 0.2× diluted BHI | B2I | 1 g/liter inulin | No. 9 | No. 10 |
| B2P | 0.5 g/liter pectin | No. 11 | No. 12 | |
| B2C | 0.5 g/liter cellulose | No. 13 | No. 14 | |
| B2M | 0.5 g/liter mucin | No. 15 | No. 16 | |
| B2S | 0.5 g/liter starch | No. 17 | No. 18 | |
| Columbia blood agar | CBA | 5% sheep’s blood | No. 19 | No. 20 |
| Chocolate agar | CHOC | Hemoglobin 2%; supplement VX | No. 21 | No. 22 |
| Tryptic soy agar | TSY | 0.5 g/liter l-cysteine hydrochloride hydrate, 10 mg/liter hemin, 1 mg/liter vitamin K, and 5 g/liter yeast extract | No. 23 | No. 24 |
| Fastidious anaerobe agar | FAA | No. 25 | No. 26 | |
| Cooked meat agar | BEEF | No. 27 | No. 28 | |
| Bifidobacterium selective medium | BSM | No. 29 | ||
| Phenyletdyl alcohol agar | PEA | 5% sheep’s blood | No. 30 | No. 31 |
| Actinomyces isolation agar | AIA | No. 32 | No. 33 | |
| Colistin-nalidixic acid agar | CNA | 5% sheep’s blood | No. 34 | No. 35 |
| McKay agar | MK | No. 36 | ||
| de Man-Rogosa-Sharpe agar | MRS | No. 37 | No. 38 | |
| Bacteroides bile esculin agar | BBE | No. 39 | No. 40 | |
| Deoxycholate agar | DOC | No. 41 | No. 42 | |
| Kanamycin-vancomycin laked blood agar | KVLB | No. 43 | ||
| Modified Gifu-anaerobic agar | MGAM | No. 44 | No. 45 | |
| Diluted MGAM | DMG | 10× diluted MGAM | No. 46 | No. 47 |
| Blautia selection medium | BLAU | MGAM + 64 μg/ml sulfamethoxazole and 16 μg/ml ciprofloxacin | No. 48 | No. 49 |
| Blood bottle-preculture + MGAM | BBMGAM | No. 50 | ||
| Blood bottle-preculture + CBA | BBCBA | No. 51 | ||
| Blood bottle-preculture + DMG | BBDMG | No. 52 | ||
| Blood bottle-preculture + BLAU | BBBLAUTb,c | No. 53 |
Methods using agars including BSM, MK, KVLB, and those preincubated in blood culture bottle (i.e., BBBLAUT, BBCBA, BBDMG, and BBMGAM) were not used under aerobic condition due to growth of rare colonies. Culture methods 1 to 43 were selected from a previous publication (16); culture methods 44 to 49 were selected from reference 17; culture methods 50 to 53 were adopted from the work of Lagier et al. (14).
BBBLAUT applied to only growing and finishing stages’ fecal microbial culture.
The blood culture bottle was a Bactec Standard Anaerobic/F culture vial (BD, Sparks, MD, USA) supplemented with 4 ml of sterile sheep blood. After microbial solution injection, the anaerobic blood culture bottle was incubated under 37°C for 4 days. Then, a volume of 50 μl of cell mixture from blood culture bottles was used for further regular anaerobic culturing on BLAU agar for 3 days.