Special considerations for conducting permeation testing of protective clothing materials with gases and chemical vapors.

Testing the permeation resistance of protective clothing materials against chemical gases and vapors requires attention to additional factors over conventional material permeation testing with liquids. Permeation testing factors relevant to gas and vapor challenges are described, and results for testing various material-gas combinations are reported. Challenging protective clothing materials with gases presents a series of special problems including gas delivery, cell integrity, sufficient analytical detection, and disposal. The concentration and other properties of gases and vapors are very sensitive to small changes in temperature and pressure. The method of delivering gases or vapors to the test cell must provide for careful regulation of these variables and maintain homogeneous contact of the chemical with the material over the test period. While many organic vapors are easily and directly detectable by gas chromatographic methods, several gases require special collection media and analytical procedures to achieve detection limits below 1 ppm. Handling of exhaust gas from the challenge chamber of the test cell must reflect safe laboratory practices without creating unnecessary chemical waste. Recommended procedures and results are presented for the six new gases added to ASTM Standard Guide F1001, Selection of Chemical Liquids and Gases to Evaluate Protective Clothing Materials, as well as for other difficult test gases used in evaluating protective clothing materials.

Turgor pressure responses of a gram-negative bacterium to antibiotic treatment, measured by collapse of gas vesicles.

The internal hydrostatic pressure of Ancylobacter aquaticus was measured by collapsing the gas vesicles with an externally applied pressure. Turgor pressure was measured in conjunction with various antibiotic treatments to elucidate some aspects of the biophysics of gram-negative cell wall function. Differences in the effects of these drugs either alone or in combination with other treatments were related to known biochemical activities of these drugs. Our previous work, demonstrating a heterogeneous cellular response to beta-lactam antibodies, was confirmed and extended. Most of the cell wall growth-inhibiting antibiotics resulted in some cells (those in component I) developing a higher pressure, while the remainder (those in component II) lost turgor. Although the fraction of the cells in each component varied a little from subculture to subculture, it did not vary with time or choice of antibiotic treatment. Mecillinam gave a nearly monophasic response. All antibiotics blocking macromolecular synthesis gave monophasic curves. The 50% collapse pressure in some cases, however, was lower higher, or the same as the control.

Variability of the turgor pressure of individual cells of the gram-negative heterotroph Ancylobacter aquaticus.

Cells of Ancylobacter aquaticus were observed under phase microscopy in a chamber to which a measured pressure could be applied. The initial collapse pressure (Ca), i.e., the lowest pressure needed to collapse the most pressure-sensitive gas vesicles, was measured for 69 cells. The cells were taken from cultures in low-density balanced exponential growth, and the experiments were performed quickly so that the bacteria were in a uniform physiological state at the time of measurement. The turgor pressure, Pt, is the difference between the pressure, C, that would cause collapse of vesicles when removed from the cell and Ca. In this paper we focus on the variability of Pt from cell to cell. Part of the observed variability of Ca was due to the variability of the collapse pressure of individual vesicles (standard deviation [SD] = 90 kPa), but because there were about 100 vesicles per cell and because a change in refracted light after the fifth vesicle (approximately) collapsed probably could be detected by the human eye, the pressure would only have an SD of 18.6 kPa due to this type of sampling error. The observed SD of Pt was 42 kPa, indicating that turgor pressure did vary considerably from cell to cell. However, the turgor pressure was independent of cell size. Statistical analysis showed that Pt would decrease 6.9 kPa over a cell cycle, but with too large an SD (19.9 kPa) to be significant. This implies that the observed change in Pt over the cell cycle is not statistically significant.

Nephelometric determination of turgor pressure in growing gram-negative bacteria.

Gas vesicles were used as probes to measure turgor pressure in Ancylobacter aquaticus. The externally applied pressure required to collapse the vesicles in turgid cells was compared with that in cells whose turgor had been partially or totally removed by adding an impermeable solute to the external medium. Since gram-negative bacteria do not have rigid cell walls, plasmolysis is not expected to occur in the same way as it does in the cells of higher plants. Bacterial cells shrink considerably before plasmolysis occurs in hyperosmotic media. The increase in pressure required to collapse 50% of the vesicles as external osmotic pressure increases is less than predicted from the degree of osmotically inducible shrinkage seen with this organism or with another gram-negative bacterium. This feature complicates the calculation of the turgor pressure as the difference between the collapse pressure of vesicles with and without sucrose present in the medium. We propose a new model of the relationship between turgor pressure and the cell wall stress in gram-negative bacteria based on the behavior of an ideal elastic container when the pressure differential across its surface is decreased. We developed a new curve-fitting technique for evaluating bacterial turgor pressure measurements.