A possible role of ProP, ProU and CaiT in osmoprotection of Escherichia coli by carnitine

Exogenously provided carnitine (beta-hydroxy-L-tau-N-trimethyl aminobutyrate) was found to stimulate aerobic growth of enterohaemorrhagic Escherichia coli O157:H7 in a medium of inhibitory osmotic strength. Its osmoprotective ability is comparable with that of betaine. As carnitine is an important compound in mammalian tissues, it is suggested that it might play a role in the growth of the pathogen on low water activity (aw) meat products. Using specific uptake mutants of E. coli K-12, it was established that, under osmotic stress, carnitine accumulates in the cytoplasm following import through the ProP and ProU transport systems. Betaine and carnitine also protect E. coli cells while growing anaerobically at inhibitory osmolarity. Under these conditions, an E. coli K-12 strain with lesions in both proP and proU accumulates low levels of L-carnitine but fails to accumulate betaine when these compounds are supplied in the external medium. This is probably a result of uptake of L-carnitine by the secondary transporter CaiT. The caiT gene forms part of the caiTABCDE operon which encodes the carnitine pathway, and is transcribed during anaerobic growth in the presence of carnitine. However, further experiments revealed that the carnitine pathway, including CaiT, does not play a significant role in osmoregulation of E. coli during anaerobiosis. Together, the results indicate that ProP and ProU are the sole transport systems involved in carnitine influx, both in aerobically and anaerobically osmotically stressed E. coli cells.     

Listeria monocytogenes repellence by enzymatically modified PES surfaces

The effect of enzyme‐catalyzed modification of poly(ethersulfone) (PES) on the adhesion and biofilm formation of two Listeria monocytogenes strains is evaluated under static and dynamic flow conditions. PES has been modified with gallic acid, ferulic acid and 4‐hydroxybenzoic acid. The surfaces modified with any of these compounds show up to 70% reduced adhesion of L. monocytogenes under static conditions and up to 95% under dynamic flow conditions compared with unmodified surfaces. Also, under static conditions the formation of biofilms is reduced by ～70%. These results indicate that the brush structures that are formed by the polymers on the PES surface directly influence the ability of microorganisms to interact with the surface, thereby reducing attachment and biofilm formation of L. monocytogenes. Based on these results, it is expected that enzyme‐catalyzed surface modification is a promising tool to reduce microbial adhesion and biofilm formation. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41576.     

The impact of oxygen availability on stress survival and radical formation of Bacillus cereus

Both the growth and stress survival of two model Bacillus cereus strains, ATCC 14579 and ATCC 10987, were tested in three different conditions varying in oxygen availability, i.e., aerobic, microaerobic and anaerobic conditions. Both B. cereus strains displayed highest growth rates and yields under aerobic conditions, whereas the microaerobic and anaerobic cultures showed similar reduced growth performances. The cells grown and exposed microaerobically and anaerobically were more resistant to heat and acid than cells that were cultured and exposed aerobically. On the other hand, the anaerobically grown cells were more sensitive to hydrogen peroxide compared to the (micro)aerobically grown cells. The increased heat- and acid-induced inactivation in aerobic conditions appeared to be associated with intracellular accumulation of excess hydroxyl and/or peroxynitrite radicals, as determined by flow cytometry in combination with the fluorescent reporter dye 3′-(p-hydroxyphenyl) fluorescein. This suggests that radical formation may contribute to inactivation of bacteria in the presence of oxygen, such as in aerobic and microaerobic conditions. No evidence was found for radical formation upon exposure to salt and hydrogen peroxide. The increased resistance to heat and acid in microaerobic and anaerobic conditions shows that oxygen availability should be taken into account when behavior of bacteria, such as B. cereus, in food industry related conditions is investigated, because oxygen availability may affect the efficiency of food preservation conditions.     

Assessment of viability of microorganisms employing fluorescence techniques

Viability assessment of microorganisms is relevant for a wide variety of applications in industry, including evaluation of inactivation treatments and quality assessment of starter cultures for beer, wine, and yoghurt production. Usually, the ability of microbial cells to reproduce is considered as the benchmark method for determination of cell viability, and this is most commonly determined by the plate count method. The time needed to form visible colonies, however, is relatively long. Therefore, there is an increasing interest in rapid methods which exploit criteria other than reproduction. In this review the applications of fluorescent probes for, e.g., determination of membrane integrity, enzyme activities, respiration, membrane potential and intracellular pH, are discussed in detail.     

Energy transduction in the thermophilic anaerobic bacterium Clostridium fervidus is exclusively coupled to sodium ions

The thermophilic, peptidolytic, anaerobic bacterium Clostridium fervidus is unable to generate a pH gradient in the range of 5.5-8.0, which limits growth of the organism to a narrow pH range (6.3-7.7). A significant membrane potential (delta psi approximately -60 mV) and chemical gradient of Na+ (-Z delta pNa approximately -60 mV) are formed in the presence of metabolizable substrates. Energy-dependent Na+ efflux is inhibited by the Na+/H+ ionophore monensin but is stimulated by uncouplers, suggesting that the Na+ gradient is formed by a primary pumping mechanism rather than by secondary Na+/H+ antiport. This primary sodium pump was found to be an ATPase that has been characterized in inside-out membrane vesicles and in proteoliposomes in which solubilized ATPase was reconstituted. The enzyme is stimulated by Na+, resistant to vanadate, and sensitive to nitrate, which is indicative of an F/V-type Na(+)-ATPase. In the proteoliposomes Na+ accumulation depends on the presence of ATP, is inhibited by the ATPase inhibitor nitrate, and is completely prevented by the ionophore monensin but is stimulated by protonophores and valinomycin. These and previous observations, which indicated that secondary amino acid transport uses solely Na+ as coupling ion, demonstrate that energy transduction at the membrane in C. fervidus is exclusively dependent on a Na+ cycle.     

The effect of the growth phase of Staphylococcus aureus on resistance to disinfectants in a suspension test.

The influence of growth phase on the resistance of Staphylococcus aureus to the surface-active agents benzalkonium chloride and dodecylbenzyl sulfonic acid and the oxidizing agents sodium hypochlorite and hydrogen peroxide was studied. The resistances of cells in different growth phases were compared to those of solid medium cells grown according to the European phase I suspension test. Using cells from different growth phases (+/- 3 x 10(7) CFU ml(-1)), we found that decline-phase cells were the most resistant cells. However, the decline-phase cell suspension contained more than 90% dead cells. A 10-fold-diluted suspension with a total concentration of cells equal to that of the other cell suspensions still revealed decline-phase cells to be generally the most resistant cell type. However, the resistance was drastically reduced, indicating that the large proportion of dead cells provided significant protection to the viable decline-phase cells. Hydrogen peroxide resistance could be partly explained by the high catalase activity in the dead-cell fraction. Exponential-phase cells were less resistant than decline-phase cells, and, surprisingly, stationary-phase cells were the least resistant of the three. Cells grown according to the European phase 1 suspension test were never the most resistant cells. Their survival was 1 to 3 log units lower than that of the most resistant cells. These findings show that the solid-medium cells currently used in disinfectant tests are not the most resistant cells that can be used.     

Elevated ovarian expression and serum concentration of alpha inhibin in the luteal phase during follicular development in the squirrel monkey (Saimiri boliviensis) compared to the human

The goal of the present investigation was to determine in the squirrel monkey the source and pattern of inhibin, a hormone known to effect reproductive steroid levels via pituitary and ovarian mechanisms. Since this seasonally polyestrous species is known to have elevated serum levels of reproductive steroids compared to other primates, the levels of ovarian alpha subunit mRNA expression and serum total alpha inhibin, estradiol, progesterone, and luteinizing hormone were measured and compared to human levels. Expression of the alpha subunit was robust in monkey luteal tissue compared to expression in human luteal tissue. Squirrel monkey serum inhibin peaked 4 days after the luteinizing hormone surge and correlated with progesterone changes. These luteal serum levels of inhibin were greater than 12 times higher than the human levels yet bio-LH activities were less than in the human during the luteal phase. Inhibin concentrations during the nonbreeding season were generally half the levels measured in the breeding season and undetectable in ovariectomized animals. However, exogenous FSH stimulation induced a marked rise in inhibin, which correlated with an estradiol rise. In conclusion, abundant alpha inhibin subunit expression in the luteal ovary of the squirrel monkey and loss of serum delectability in ovariectomized animals indicates that the principle source of inhibin in the squirrel monkey is the ovary. Elevated serum inhibin levels during the luteal phase concurrent with ovulatory-size follicular development is unique among species studied thus far. Possible simultaneous inhibin production from both follicular and luteal tissue may be responsible for the exceptionally high inhibin levels.     

Antimicrobial lysozyme-containing starch microgel to target and inhibit amylase-producing microorganisms

The aim of this study is to determine the release of lysozyme from oxidized starch microgels and subsequently test its antimicrobial activity. The gels are made of oxidized potato starch polymers, which are chemically cross-linked by sodium trimetaphosphate (STMP). The microgel is negatively charged and interacts with positively charged lysozyme by electrostatic attraction. Application of the lysozyme-containing starch particles to environments contaminated with microbes, may lead to hydrolysis of the starch by microbial enzymes. As a result, lysozyme is released in the environment where it inhibits microbial growth. In this study, first bacteria were screened for amylase production and lysozyme sensitivity. Then, the bacteria were mixed with empty gel particles (i.e., without lysozyme) in a Nutrient Broth liquid medium to test whether the bacteria that can produce amylase are also able to degrade oxidized starch gel. Subsequently the amylase-producing lysozyme sensitive bacteria, Bacillus licheniformis 7558 and Bacillus subtilis 168, were selected for further quantification of the antimicrobial activity of the gel-lysozyme particles after incubation with these bacteria in Nutrient Broth liquid suspensions. The results prove that the starch microgel has a potential as antimicrobial carrier targeting amylase-producing and lysozyme-sensitive bacteria. The controlled antimicrobial delivery for inactivating undesired microorganisms may find applications in food related systems, where amylase-producing bacteria may be abundantly present.     

Antibacterial activity of carbon nanoparticles isolated from chimney soot

Carbon nanoparticles (CNPs) are isolated from chimney soot and characterised by various tools such as X‐ray diffraction, scanning electron microscopy, transmission electron microscopy and ultraviolet–visible spectroscopy. The X‐ray diffraction studies confirm the presence of C₆₀ nanoparticles in the isolated sample. The thermal properties of the prepared CNPs are recorded using thermogravimetric analysis and differential thermal analysis. The analysis of the antibacterial activity of the synthesised CNPs against selected Gram‐positive and Gram‐negative bacterial strains is also investigated. The systematic study confirms that CNPs collected from chimney soot exhibit good antibacterial potency against Staphylococcus aureus, Streptococcus pyogenes, Klebsiella pneumoniae, and Proteus mirabilis.Inspec keywords: ultraviolet spectra, scanning electron microscopy, visible spectra, differential thermal analysis, thermal analysis, antibacterial activity, nanoparticles, X‐ray diffraction, nanofabrication, transmission electron microscopy, carbonOther keywords: chimney soot, transmission electron microscopy, ultraviolet–visible spectroscopy, thermal properties, thermogravimetric analysis, differential thermal analysis, antibacterial activity, carbon nanoparticles, X‐ray diffraction study, gram‐positive bacterial strains, gram‐negative bacterial strains, antibacterial potency, scanning electron microscopy, C₆₀      

Characterization of two phosphate transport systems in Acinetobacter johnsonii 210A

The transport of P(i) was characterized in Acinetobacter johnsonii 210A, which is able to accumulate an excessive amount of phosphate as polyphosphate (polyP) under aerobic conditions. P(i) is taken up against a concentration gradient by energy-dependent, carrier-mediated processes. A. johnsonii 210A, grown under P(i) limitation, contains two uptake systems with Kt values of 0.7 +/- 0.2 microM and 9 +/- 1 microM. P(i) uptake via the high-affinity component is drastically reduced by N,N'-dicyclohexylcarbodiimide, an inhibitor of H(+)-ATPase, and by osmotic shock. Together with the presence of P(i)-binding activity in concentrated periplasmic protein fractions, these results suggest that the high-affinity transport system belongs to the group of ATP-driven, binding-protein-dependent transport systems. Induction of this transport system upon transfer of cells grown in the presence of excess P(i) to P(i)-free medium results in a 6- to 10-fold stimulation of the P(i) uptake rate. The constitutive low-affinity uptake system for P(i) is inhibited by uncouplers and can mediate counterflow of P(i), indicating its reversible, secondary nature. The presence of an inducible high-affinity uptake system for P(i) and the ability to decrease the free internal P(i) pool by forming polyP enable A. johnsonii 210A to reduce the P(i) concentration in the aerobic environment to micromolar levels. Under anaerobic conditions, polyP is degraded again and P(i) is released via the low-affinity secondary transport system.