Table 3.
Physiological characterization of N. vectensis associated strains selected for genome-sequencing.
| Strain ID | Medium preference | Growth condition | ||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Aerobic growth (1) | Microaerophilic growth (2) | (3) | Temperature (5) | Salinity (6) | pH (7) | |||||||||||||||||
| 16SrRNA top BlastN | Isolation Date/Origin | LB | TSB | 2216 | Na2S | Na2S2O3 | 2216 | C-W | Bruc. blood | AB resistance | Cell morphology (1) | Colony morphology (4) | 4°C | 16°C | 22°C | 28°C | 37°C | 45°C | Catalase | Gram Staining | ||
| Po-Gab—P. oleovorans subsp. oleovorans (98.2%) | Isolated from lab anemones (2/2010). Gentamycin treatment after tissue homogenization; strain was isolated on 2216 marine agar | +++ | +++ | + | − | − | + | +++ | +++ | NAR, CmR, ApR | Motile rod 1~1.5 × 0.5 μm | 1 mm, ivory, opaque, circular, entire margin, bumpy surface | − | + | ++ | +++ | +++ | + | 0–5% (max. 2–3%) | pH6–10 (max. pH7) | ++ | − |
| Po-ls—P. oleovorans subsp. oleovorans (98.2%) | Isolated from lab anemones (2/2010). Enriched in sulfide-gradient-tube with ASW and 0.1 mM NaN03; isolation on 2216 marine agar | +++ | +++ | + | − | − | + | +++ | +++ | NAR, CmR, ApR | Motile rod 1~1.5 × 0.5 μm | 1 mm, ivory, opaque, circular, entire margin, bumpy surface | − | + | ++ | +++ | +++ | + | 0–5% (max. 2–3%) | pH6–10 (max. pH7) | ++ | − |
| Po-B4—P. oleovorans subsp. oleovorans (98.2%) | Isolated from lab anemones (1/2008). Homogenate plated on 2216 marine agar | +++ | +++ | + | − | − | + | +++ | +++ | NAR, CmR, ApR | Motile rod 1~1.5 × 0.5 μm | 1 mm, translucent, ivory, circular bumpy surface, entire margin | − | + | ++ | +++ | +++ | + | 0–5% (max. 2–3%) | pH6–10 (max. pH7) | ++ | – |
| Po-47—P. oleovorans subsp. oleovorans) (98.2%) | Isolated on 2216 marine agar from homogenized laboratory-raised anemones obtained from John Finnerty's Lab (BU) (11/2009) | +++ | +++ | + | − | − | + | +++ | +++ | NAR, CmR | Motile rod 1~1.5 × 0.5 μm | 0.5–0.7 mm, translucent, ivory, circular with entire margin, smooth shiny surface | − | + | ++ | +++ | +++ | + | 0–5% (max. 2–3%) | pH6–10 (max. pH7) | ++ | − |
| Lt-F1—Limnobacter thiooxidans (98.9%) | Field animals were used as innocula (4/2010); isolated on 0.1X strength 2216 marine broth solidified with gellan gum | +, 72 h | −, 72 h | +, 72 h | − | − | + | − | + | None observed | Motile rod 1~1.5 × 0.5 μm | 0.5 mm ivory, translucent, circular entire margine, smooth and shiny surface | − | + | + | + | + | − | 1–2% (max. 2%) | pH6–10 (max. pH8) | ++ | − |
| Lt-FCMA—Limnobacter thiooxidans (98.9%) | Field animals were used as innocula (4/2010); Collagenase was used for tissue maceration; isolated on 2216 marine agar | ++, 72 h | −, 72 h | +, 72 h | − | − | + | − | + | None observed | Motile rod 1~1.5 × 0.5 μm | 1 mm ivory, translucent/transparent, circular, umbonate, shiny/smooth surface | − | − | + | ++ | ++ | − | 0–3% (max. 2%) | pH6–10 (max. PH7) | ++ | – |
| Rr-ls—Rhizobium radiobacter (97.7%) | Lab anemone inocula (2/2010); Enriched in sulfide-gradient-tube with ASW and 0.4mM NaN03; further isolation on Brucella Blood agar | +++ | +++ | + | − | − | + | +++ | ++ | NAR, SmR, CmR, KnR | Short and stout rods 1.5 × 0.7 μm, relatively less motile, clusters of 10–20 cells. | Pinpoint, ivory, opaque, circular smooth with entire margin | − | + | ++ | +++ | + | − | 0–3% (max. 2%) | pH6–10 (max. pH7) | ++ | − |
| Rr-D8—Rhizobium radiobacter (97.7%) | Isolated on 2216 marine agar from homogenized laboratory-raised anemones (1/2008) | +++ | +++ | ++ | − | − | ++ | +++ | ++ | SmR | Slender rods 2–2.5 × 0.7 μm, relatively less motile and occur in pairs | Pinpoint, ivory, opaque, circular smooth surface, entire margin | − | + | ++ | +++ | + | + | 0–3% (max. 2%) | pH6–10 (max. pH7) | ++ | − |
| Rr-D5—Rhizobium radiobacter (97.7%) | Isolated on 2216 marine agar from homogenized laboratory-raised anemones (1/2008) | +++ | +++ | ++ | − | − | + | +++ | +++ | NAR, SmR, CmR, KnR, ApR | Short and stout, motile rods 0.7–1 × 0.5–0.7 μm in size | Pinpoint, ivory, opaque, circular smooth surface, entire margin | − | + | ++ | +++ | +++ | + | 0–7% (max. 2–3%) | pH6–10 (max. PH7) | ++ | − |
| Ss-F1—Stappia stellulata (98.7%) | Field animals were used as innocula (4/2010); isolated on 0.1X strength Marine (= 2216) broth solidified with gellan gum | +++ | +++ | ++ | − | − | ++ | +++ | +++ | NAR, SmR | Motile rod 1.5–2 × 0.5 μm | Pinpoint, ivory, opaque, circular smooth surface with entire margin | −− | ++ | +++ | +++ | +++ | + | 2–5% (max. 3%) | pH6–10 (max. PH7) | ++ | − |
+++, very good growth; ++, good growth; +, weak growth; –, no growth. (1) Growth under aerobic conditions: All strains were shaken at 28°C for 48 h unless otherwise specified. (2) Growth under Microaerophilic conditions. All strains were incubated with GasPak EZ Anaerobic Pouch System (BD) at 28°C for 3 day; MM, Minimal Media; C-W, Campylobacter Wollinella Agar; Bruc. Blood, Brucella Blood Agar. (3) Antibiotic Resistance. R, Resistant; NA, 4 ppm Nalidixic acid; Sm, 100 ppm Streptomycin; Cm, 10 ppm Chloramphenicol; Kn, 100 ppm Kanamycin; and Ap, 100 ppm Ampicillin. (4) Colony morphology determined after growth on 2216 agar at 28°C for 2 days unless otherwise specified (5) All strains were grown on 2216 plate and observed after 4 days. (6) All strains were grown on LB broth omitting NaCl at 28°C for 3 days. (7) All strains were grown on 2216 broth buffered with acetic acid, NaH2PO4, or Tris base at 28°C for 48 h.