This paper reports the development of microencapsulated bacteriophage Felix O1 for oral delivery using a chitosan-alginate-CaCl 2 system. In vitro studies were used to determine the effects of simulated gastric fluid (SGF) and bile salts on the viability of free and encapsulated phage. Free phage Felix O1 was found to be extremely sensitive to acidic environments and was not detectable after a 5-min exposure to pHs below 3.7. In contrast, the number of microencapsulated phage decreased by 0.67 log units only, even at pH 2.4, for the same period of incubation. The viable count of microencapsulated phage decreased only 2.58 log units during a 1-h exposure to SGF with pepsin at pH 2.4. After 3 h of incubation in 1 and 2% bile solutions, the free phage count decreased by 1.29 and 1.67 log units, respectively, while the viability of encapsulated phage was fully maintained. Encapsulated phage was completely released from the microspheres upon exposure to simulated intestinal fluid (pH 6.8) within 6 h. The encapsulated phage in wet microspheres retained full viability when stored at 4°C for the duration of the testing period (6 weeks). With the use of trehalose as a stabilizing agent, the microencapsulated phage in dried form had a 12.6% survival rate after storage for 6 weeks. The current encapsulation technique enables a large proportion of bacteriophage Felix O1 to remain bioactive in a simulated gastrointestinal tract environment, which indicates that these microspheres may facilitate delivery of therapeutic phage to the gut.

Background: In order to study the mechanism of U(VI) reduction, the effect of deleting c-type cytochrome genes on the capacity of Geobacter sulfurreducens to reduce U(VI) with acetate serving as the electron donor was investigated.

This manuscript is dedicated to our friend, mentor, and coauthor Dr Terry Beveridge, who devoted his scientific career to advancing fundamental aspects of microbial ultrastructure using innovative electron microscopic approaches. During his graduate studies with Professor Robert Murray, Terry provided some of the first glimpses and structural evaluations of the regular surface arrays (S-layers) of Gram-negative bacteria (Beveridge & Murray, 1974, 1975, 1976a). Beginning with his early electron microscopic assessments of metal binding by cell walls from Gram-positive bacteria (Beveridge & Murray, 1976b, 1980) and continuing with more than 30 years of pioneering research on microbe-mineral interactions (Hoyle & Beveridge, 1983, 1984; Ferris et al., 1986; Gorby et al., 1988; Beveridge, 1989; Mullen et al., 1989; Urrutia Mera et al., 1992; Mera & Beveridge, 1993; Brown et al., 1994; Konhauser et al., 1994; Beveridge et al., 1997; Newman et al., 1997; Lower et al., 2001; Glasauer et al., 2002; Baesman et al., 2007), Terry helped to shape the developing field of biogeochemistry. Terry and his associates are also widely regarded for their research defining the structure and function of outer membrane vesicles from Gram-negative bacteria that facilitate processes ranging from the delivery of pathogenic enzymes to the possible exchange of genetic information. The current report represents the confluence of two of Terry's thematic research streams by demonstrating that membrane vesicles produced by dissimilatory metal-reducing bacteria from the genus Shewanella catalyze the enzymatic transformation and precipitation of heavy metals and radionuclides. Under low-shear conditions, membrane vesicles are commonly tethered to intact cells by electrically conductive filaments known as bacterial nanowires. The functional role of membrane vesicles and associated nanowires is not known, but the potential for mineralized vesicles that morphologically resemble nanofossils to serve as palaeontological indicators of early life on Earth and as biosignatures of life on other planets is recognized.

Electron microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis with silver staining and 1 H, 13 C, and 31 P-nuclear magnetic resonance (NMR) were used to detect and characterize the lipopolysaccharides (LPSs) of several Shewanella species. Many expressed only rough LPS; however, approximately one-half produced smooth LPS (and/or capsular polysaccharides). Some LPSs were affected by growth temperature with increased chain length observed below 25°C. Maximum LPS heterogeneity was found at 15 to 20°C. Thin sections of freeze-substituted cells revealed that Shewanella oneidensis, S. algae, S. frigidimarina, and Shewanella sp. strain MR-4 possessed either O-side chains or capsular fringes ranging from 20 to 130 nm in thickness depending on the species. NMR detected unusual sugars in S. putrefaciens CN32 and S. algae BrY DL . It is possible that the ability of Shewanella to adhere to solid mineral phases (such as iron oxides) could be affected by the composition and length of surface polysaccharide polymers. These same polymers in S. algae may also contribute to this opportunistic pathogen's ability to promote infection.Shewanella organisms are generally associated with aquatic habitats and play important roles in the cycling of particulate iron and organic matter, but they can also be opportunistic pathogens (33,45). Because of their environmental significance, they are presently under vigorous investigation and many new species were recently included in the genus (45). Most Shewanella organisms are capable of dissimilatory reduction of a wide range of electron acceptors, including metal oxides [e.g., those of Fe(III) and Mn(IV)]. Several reductive mechanisms are possible. Electron flux from the organism to the solid oxide may occur via (i) direct contact of bacterial outer membrane and oxide surface (2, 9, 21, 31), (ii) organic shuttles (such as humic acids or quinones to mediate electron flow (22, 32), or (iii) a combination of both processes. For the direct contact model, metal reductases (such as c-type cytochromes) seem to be embedded in the outer membrane of dissimilatory metal-reducing bacteria to facilitate electron flow (13,29,30). Lipopolysaccharide (LPS) and outer membrane proteins could play principal roles in establishing and maintaining contact with oxide minerals so that electron transport to the terminal acceptor occurs. The junction between cell and mineral must be tight so as to ensure that the reductase functions effectively (23). However, the cell surface structure and LPS of Shewanella, which could affect the cell-mineral connection, are poorly understood (28,39,40,48). In this present article we characterize those structural elements that can extend beyond the outer face of the outer membrane, i.e., the LPS O-side chains and capsular polymers.Bacterial strains and growth conditions. The strains used in this study are shown in Table 1. Most of these strains were kindly provided by Doug Lies (Jet Propulsion Laboratory, Pasadena, Calif.). S. algae BrY was supplied by both D. Lies an...

Shewanella strains have previously been studied with regard to their cell surface ultrastructure and LPS composition. They have now been further characterized with respect to their surface physicochemistry and ability to adhere to haematite. The surfaces of the Shewanella strains were found to be electronegative and hydrophilic, and these properties could be correlated with LPS composition or the presence of capsular polysaccharides. Strains expressing rough LPS with no capsule were more hydrophobic and electronegative than those possessing smooth LPS or capsules. By combining different approaches, such as contact-angle measurement, hydrophilic/ hydrophobic chromatography, microelectrophoresis, adhesion assays and calculation of interaction energies, it was shown that electrostatic interactions predominate over hydrophobic interactions at the cell-iron oxide interface. Bacterial adhesion to haematite was significantly reduced in strains expressing smooth LPS or a capsule. These findings remained true for Shewanella strains grown under either aerobic or anaerobic conditions, although the surfaces of anaerobic cells appeared to be less electronegative and more hydrophilic than those of aerobic cells.