The PilQ secretin from the pathogenic bacterium Neisseria meningitidis is an integral outer membrane protein complex which plays a crucial role in the biogenesis of type IV pili. We present here the first three-dimensional structure of this type of secretin at 2.5-nm resolution, obtained by single-particle averaging methods applied to the purified protein complex visualized in a negative stain. In projection, the PilQ complex is circular, with a donut-like appearance. When viewed from the side it has a rounded, conical profile. The complex was demonstrated to have 12-fold rotational symmetry, and this property was used to improve the quality of the density map by symmetry averaging. The dominant feature of the structure is a cavity, 10 nm deep, within the center of the molecule. The cavity is funnel-shaped in cross section, measures 6.5 nm in diameter at the top of the complex, and tapers to a closed point, effectively blocking formation of a continuous pore through the PilQ complex. These results suggest that the complex would have to undergo a conformational change in order to accommodate an assembled pilus fiber of diameter 6.5 nm running through the outer membrane.
This study investigates different UV doses (mJ/cm(2)) and the effect of dark incubation on the survival of the algae Tetraselmis suecica, to simulate ballast water treatment and subsequent transport. Samples were UV irradiated and analyzed by flow cytometry and standard culturing methods. Doses of ≥400 mJ/cm(2) rendered inactivation after 1 day as measured by all analytical methods, and are recommended for ballast water treatment if immediate impairment is required. Irradiation with lower UV doses (100-200 mJ/cm(2)) gave considerable differences of inactivation between experiments and analytical methods. Nevertheless, inactivation increased with increasing doses and incubation time. We argue that UV doses ≥100 mJ/cm(2) and ≤200 mJ/cm(2) can be sufficient if the water is treated at intake and left in dark ballast tanks. The variable results demonstrate the challenge of giving unambiguous recommendations on duration of dark incubation needed for inactivation when algae are treated with low UV doses.
UV] irradiation, heat, and cavitation) treatment technologies (Shannon et al. 2008, Werschkun et al. 2012, 2014). Traditionally, water analysis used to assess most treatment technologies has depended on cultivation, such as the plate count or the most probable number (MPN) technique. Cultivation methods measure viability of organisms present, i.e. the ability of a cell to reproduce. Vital (live) cells can be either viable or non-viable, whereas non-vital (dead)
The transfer of non-native, possibly invasive species in ship's ballast water is of global concern, and the International Maritime Organization and U.S. Coast Guard have adopted standards to minimize the environmental footprint caused by the maritime industry. In this study, seawater spiked with the phytoplankter Tetraselmis suecica, was treated with Knutsen Ballast Water Treatment Technology (KBAL), combining UV irradiation with an in-line vacuum drop. The test water was subsequently incubated in dark tanks, simulating what happens onboard a ship, where ballast water is treated at intake, stored in dark ballast tanks during the voyage, and then treated at discharge. Our results of the test water treated with KBAL and stored 5 days in the dark showed < 10 viable T. suecica cells ml −1 when assessing reproduction and > 10 living cells ml −1 when assessing metabolism. This highlights the challenge UV-based BWTS can encounter when meeting testing regimes assessing different characteristics of life. By comparing the effects caused by KBAL treatment with effects caused by UV irradiation only, we demonstrated that the pressure/vacuum technology seems to improve the disinfection effect. In addition, our investigations point out possible challenges with in situ conditions getting representative ballast water samples.
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