The Microaerophile SPirillum volutans : Cultivation on Complex Liquid and Solid Media

Abstract

Spirillum volutans grows only under microaerobic conditions in a-peptonesuccinate-salts broth, but can grow aerobically when the peptone is replaced by vitamin-free acid-hydrolyzed casein broth. The addition of potassium metabisulfite, norepinephrine, catalase or superoxide dismutase (SOD) permitted aerobic growth in peptone-succinate-salts broth. A combination of catalase and SOD had a synergistic effect. S. volutans lacked catalase and had only a low level of peroxidase activity, but did possess SOD activity (12 to 14 U/mg of protein). The organism was found to be extraordinarily sensitive to exogenous hydrogen peroxide. Illumination of peptone-succinate-salts broth generated hydrogen peroxide and rendered the medium inhibitory to growth. A combination of catalase and SOD prevented this inhibition. Growth of S. volutans on solid media, not previously possible, was accomplished by the use of vitamin-free acid-hydrolyzed casein and peptone-succinate-salts agar media; maximum growth responses were dependent on the following combination of factors: addition of bisulfite, catalase, or SOD, protection of the media from illumination, incubation in a highly humid atmosphere, and incubation under atmospheres of 12% oxygen or less. The results indicate that the microaerophilic nature of S. volutans is attributable largely to the high sensitivity of the organism to exogenous hydrogen peroxide and, to a lesser extent, superoxide radicals occurring in the culture medium.

Detection of Actinobacteria cultivated from environmental samples reveals bias in universal primers

“…Many environmental factors contribute to the inability of micro-organisms to replicate on laboratory media. In particular, the accumulation of reactive oxygen species (ROS) in the growth media has been shown to be deleterious to the recovery of injured, stressed, or microaerophilic bacteria, while the addition of ROS-degrading compounds has been shown to support the cultivation of these micro-organisms (Burton and Morita 1964;Kautter 1976, 1977;Martin et al 1976;Padgett et al 1982;Cox et al 1990;Hood et al 1990;Olson et al 2000;Stevenson et al 2004). These studies suggest that the addition of ROS-degrading compounds to culture media may permit the cultivation of a greater percentage of the environmental microbial community than is currently contained within repositories.…”

Identification of superoxide dismutase activity in Borrelia burgdorferi

“…E. coli and aerobes such as Pseudomonas aeruginosa (25) contain specific activities of SOD that are severalfold greater than those of obligate anaerobes (26). The SOD levels in B. burgdorferi were found to be intermediate between those of the aerobes and those of the anaerobes and most like those of microaerophiles such as Campylobacter jejuni (30), Helicobacter pylori (43), and Spirillum volutans (36).…”

Sensitivity of an Oligotrophic Lake Planktonic Bacterial Community to Oxygen Stress

“…Superoxide dismutase and catalase were added to the standard medium in one experiment to determine whether superoxide or hydrogen peroxide was being generated exogenously to cells in inhibitory concentrations. Whereas positive evidence for such exogenous generation exists for the microaerophiles Spirillum volutans and Campylobacter fetus (3,7), the same enzyme concentrations had no effect (a > 0.05) on the Mountain Lake community treated with or without the enzymes, as measured by CFU development and 14C incorporation into total lipid (Table 4). Endogenous generation and activity of superoxide and hydrogen peroxide remains a possibility.…”