Search

The availability of water, which can be expressed in terms of water activity (a(w)), is one of the most important determinants for microbial homeostasis and growth on surface to air interfaces. Here we show, using an environmental control chamber containing a precisely controlled temperature/a(w) gradient in combination with a mathematical approach, that the environmental a(w) growth limit of a microorganism can be lower than its intracellular a(w) limit. This internal limit represents the point at which microbial cells cannot lower their internal a(w) any further in response to low external a(w) values without interfering with essential intracellular processes. To grow at external a(w) values below their internal limit, microbes need to generate more water metabolically than they lose to their environment. This internal a(w) limit can be calculated by measuring the a(w) growth limit of an organism at different water vapour diffusivities using barometric pressure as a variable. Fascinating morphological changes, such as rope-like superstructures formed by B. subtilis, are furthermore observed in response to low external a(w) values in particular around the calculated intracellular a(w) limit. The intracellular a(w) limit of an organism is a decisive parameter for water limitation-induced adaptations in cellular hydrophilicity and morphogenesis., (© 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.)

The response of different bacterial species to reduced water availability was studied using a simple relative humidity gradient technique. Interestingly, distinct differences in morphology and growth patterns were observed between populations of the same species growing at different relative humidity. Gram-positive cocci increased in cell size as they approached humidity growth limits and staphylococcal species started growing in tetrad/cubical formations instead of their normal grape-like structures. Gram-negative rods displayed wave-like patterns, forming larger waves as they became increasingly filamentous at low humidity. In contrast, cells of the Gram-positive bacterium Bacillus subtilis became shorter, curved, and eventually almost coccoid. Moreover, B. subtilis started to sporulate at low humidity. The altered morphology and/or growth patterns of bacteria growing at low humidity might be more ecologically relevant than their textbook appearance at high humidity since their natural habitats are often dry. Transmission electron microscopic analyses revealed that staphylococci grown at low humidity have significantly thickened cell walls, which may explain why these cells displayed increased resistance to vancomycin. We conclude that our relative humidity gradient technique is widely applicable for investigating effects of relative humidity on microbial survival, growth and competitive success at solid-air interfaces, making it a versatile tool in microbial ecology.

Many of the substrates available as energy sources for microorganisms in the human colon, including dietary plant fibre and secreted mucin, are insoluble. It seems likely that such insoluble substrates support a specialized microbiota, and in order to test this hypothesis, faecal samples from four healthy subjects were fractionated into insoluble (washed particulate) and liquid fractions. Analysis of 1252 PCR-amplified 16S rRNA sequences revealed a significantly lower percentage of Bacteroidetes (P = 0.021) and a significantly higher percentage of Firmicutes (P = 0.029) among bacterial sequences amplified from particle-associated (mean 76.8% Firmicutes, 18.5% Bacteroidetes) compared with liquid phase (mean 65.8% Firmicutes, 28.5% Bacteroidetes). Within the Firmicutes, the most significant association with solid particles was found for relatives of Ruminococcus-related clostridial cluster IV species that include Ruminococcus flavefaciens and R. bromii, which together accounted for 12.2% of particle-associated, but only 3.3% of liquid phase, sequences. These findings were strongly supported by microscopy, using group-specific FISH probes able to detect these species. This work suggests that the primary colonizers of insoluble substrates found in the gut are restricted to certain specialized groups of bacteria. The abundance of such primary degraders may often be underestimated because of the difficulty in recovering these bacteria and their nucleic acids from the insoluble substrate.