TL;DR: This study compared the growth of H. pylori in vitro, under various gas atmospheres, and determined some specific changes in the physiology of bacteria grown under different oxygen partial pressures.
Abstract: Background: There is no general consensus about the specific oxygen and carbon dioxide requirements of the human pathogen Helicobacter pylori. This bacterium is considered a microaerophile and consequently, it is grown under atmospheres at oxygen tensions 5–19% and carbon dioxide tensions 5–10%, both for clinical and basic and applied research purposes. The current study compared the growth of H. pylori in vitro, under various gas atmospheres, and determined some specific changes in the physiology of bacteria grown under different oxygen partial pressures.
Methods: Measurements of bacterial growth under various conditions were carried out employing classical solid and liquid culture techniques. Enzymatic activities were measured using spectrophotometric assays.
Results: H. pylori and all the other Helicobacter spp. tested had an absolute requirement for elevated carbon dioxide concentrations in the growth atmosphere. In contrast with other Helicobacter spp., H. pylori can tolerate elevated oxygen tensions when grown at high bacterial concentrations. Under 5% CO2, the bacterium showed similar growth in liquid cultures under oxygen tensions from microaerobic (< 5%) to fully aerobic (21%) at cell densities higher than 5 × 105 cfu/ml for media supplemented with horse serum and 5 × 107 cfu/ml for media supplemented with β-cyclodextrin. Evidence that changes occurred in the physiology of H. pylori was obtained by comparing the activities of ferredoxin:NADH (nicotinamide adenine dinucleotide) oxidoreductases of bacteria grown under microaerobic and aerobic atmospheres.
Conclusions: H. pylori is a capnophile able to grow equally well in vitro under microaerobic or aerobic conditions at high bacterial concentrations, and behaved like oxygen-sensitive microaerophiles at low cell densities. Some characteristics of H. pylori cells grown in vitro under microaerobic conditions appeared to mimic better the physiology of organisms grown in their natural niche in the human stomach.
TL;DR: The ability to culture an environmental isolate indicates that organisms found in CO2 environments from previous genomic and metagenomics studies are viable, metabolizing, and potentially affecting the surrounding environment.
Abstract: When CO2 is sequestered into the deep subsurface, changes to the subsurface microbial community will occur. Capnophiles, microorganisms that grow in CO2-rich environments, are some organisms that may be selected for under the new environmental conditions. To determine whether capnophiles comprise an important part of CO2-rich environments, an isolate from Crystal Geyser, Utah, U.S.A., a CO2- rich spring considered a carbon sequestration analogue, was characterized. The isolate was cultured under varying CO2, pH, salinity, and temperature, as well as different carbon substrates and terminal electron acceptors (TEAs) to elucidate growth conditions and metabolic activity. Designated CG-1, the isolate is related (99%) to Lactobacillus casei in 16S rRNA gene identity, growing at PCO2 between 0 to 1.0 MPa. Growth is inhibited at 2.5 MPa, but stationary phase cultures exposed to this pressure survive beyond 5 days. At 5.0 MPa, survival is at least 24 hours. CG-1 grows in neutral pH, 0.25 M NaCl, and between 25° to 45°C andconsumes glucose, lactose, sucrose, or crude oil, likely performing lactic acid fermentation. Fatty acid profiles between 0.1 MPa to 1.0 MPa suggests decreases in cell size and increases in membrane rigidity. Transmission electron microscopy reveals rod shaped bacteria at 0.1 MPa. At 1.0 MPa, cells are smaller, amorphous, and produce abundant capsular material. Its ability to grow in environments regardless of the presence of CO2 suggests we have isolated an organism that is more capnotolerant than capnophilic. Results also show that microorganisms are capable of surviving the stressful conditions created by the introduction of CO2 for sequestration. Furthermore, our ability to culture an environmental isolate indicates that organisms found in CO2 environments from previous genomic and metagenomics studies are viable, metabolizing, and potentially affecting the surrounding environment.