François Krier

Bovine hemoglobin : An attractive source of antibacterial peptides

A peptic hemoglobin hydrolysate was fractioned by a semi-preparative reversed-phase HPLC and some fractions have an antibacterial activity against four bacteria strains: Micrococcus luteus A270, Listeria innocua, Escherichia coli and Salmonella enteritidis. These fractions were analyzed by ESI/MS and ESI/MS/MS, in order to characterize the peptides in these fractions. Each fraction contains at least three peptides and some fractions contain five peptides. All these fractions were purified several times by HPLC to obtain pure peptides. Thirty antibacterial peptides were identified. From the isolated antibacterial peptides, 24 peptides were derived from the alpha chains of hemoglobin and 6 peptides were derived from the beta chains of hemoglobin. The lowest concentration of these peptides (minimum inhibitory concentration (MIC)) necessary to completely inhibit the growth of four bacteria strain was determined. The cell population of all of the tested bacteria species decreased by at least 97% after a 24-h incubation with any of the peptides at the minimum inhibitory concentration.

New antibacterial peptide derived from bovine hemoglobin

Peptic digestion of bovine hemoglobin at low degree of hydrolysis yields an intermediate peptide fraction exhibiting antibacterial activity against Micrococcus luteus A270, Listeria innocua, Escherichia coli and Salmonella enteritidis after separation by reversed-phase HPLC. From this fraction a pure peptide was isolated and analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). This peptide correspond to the 107-136 fragment of the alpha chain of bovine hemoglobin. The minimum inhibitory concentrations (MIC) towards the four strains and its hemolytic activity towards bovine erythrocytes were determined. A MIC of 38 microM was reported against L. innocua and 76 microM for other various bacterial species. This peptide had no hemolytic activity up to 380 microM concentration.

Isolation and characterization of four antibacterial peptides from bovine hemoglobin.

Peptic digestion of bovine hemoglobin at low degree of hydrolysis yields several intermediate peptide fractions after separation by reversed phase HPLC exhibiting antibacterial activity against Micrococcus luteus A270, Listeria innocua, Escherichia coli, and Salmonella enteritidis. From these fractions, four new antibacterial peptides were isolated and analyzed by ESI-MS/MS. Three of these peptides correspond to fragments of the alpha-chain of bovine hemoglobin: alpha107-141, alpha137-141, and alpha133-141, and one peptide to the beta-chain: beta126-145. The minimum inhibitory concentrations (MIC) of these peptides towards the four strains and their hemolytic activity towards bovine erythrocytes were determined.

Involvement of cell shape and flagella in the bacterial retention during percolation of contaminated water through soil columns in tropical region.

Microorganisms’ retention in soil contributes to the natural purification of groundwater. Bacteria found in groundwater are generally of various shapes. The aim of this study was to assess the importance of cell shape and flagella in bacterial retention during polluted water percolation through two soil columns CA and CB, in the equatorial region in Central Africa. Percolation tests were carried out using different water loads samples which were contaminated by Escherichia coli (straight rods, peritrichous flagella), Vibrio parahaemolyticus (rods bacteria, polar flagella), and Staphylococcus saprophyticus (spherical, free-flagellum). It has been noted that showed that through soil column CA, the mean values of cells retention ratios (TR ) varied with bacteria species considered, and from one applied water load sample to another. E. coli T R and that of S. saprophyticus were not significantly different (P> 0.05) for the two soil columns. V. parahaemolyticus TR significantly differed from that of E. coli and S. saprophyticus through soil column CA (P< 0.01) when the highest water load was applied, and through soil column CB (P< 0.05) for each of water load applied. A relative hierarchical arrangement of retained cells based on the TR showed that V. parahaemolyticus was less retained through the 2 soil columns. S. saprophyticus in most cases was more retained than others. The physical properties of the bacterial cell must be taken into consideration when evaluating the transfer of bacteriological pollutants towards groundwater.

Assessment of the hierarchical involvement of chemical characteristics of soil layer particles during bacterial retention in Central Africa

The main purpose of this investigation was to rank, in hierarchical order, some soil chemical factors that impact the sorption parameters of fecal indicator bacteria and Salmonella sp, during their transfer from soil surface to the groundwater table. Sorption coefficients (Kf) and linearity exponents (Ϫ) were obtained using the Freundlich isotherm. When Salmonella sp was present alone, the values of Kf varied from 2.70 log units mg−1 to 7.84 log units mg−1 of soil, and Ϫ varied from 0.04 to 2.70. When the two fecal indicator bacteria were present, Kf varied from 0.48 log units mg−1 to 8.53 log units mg−1 of soil, and Ϫ varied from 0.06 to 1.19. When fecal indicator bacteria and Salmonella sp were mixed, Kf varied from 1.73 log units mg−1 to 7.70 log units mg−1 of soil, and Ϫ varied from 0.09 to 0.88. The predominant factors controlling each sorption parameter varied with respect to soil layer and the bacterial species involved.

Photoinactivation of Staphylococcus aureus and Vibrio parahaemolyticus in the model aquatic microcosm: effect of light intensity and dissolved biodegradable organic compound

The impact of light (1,000 × 100,000 lx) on the inactivation of S. aureus and V. parahaemolyticus has been assessed under different concentrations of dissolved biodegradable organic compound (BOC) at pH 7.0. First, a gradual decrease in the number of cultivable cells was observed. Secondly, a cell reactivation was observed and it was marked in the absence of BOC. In the absence of BOC, the lowest value of cell inhibition rate (CIR) during the first 3 h was 0.138 h(-1) for S. aureus and 0.218 h(-1) for V. parahaemolyticus. In the presence of 10,100 and 1,000 mg/l of BOC, it was 0.196 h(-1), 0.243 h(-1) and 0.257 h(-1) for S. aureus respectively, and 0.285 h(-1), 0.306 h(-1) and 0.409 h(-1) for V. parahaemolyticus respectively. The CIRs values of each bacterial species significantly varied (P<0.001) with the changes in BOC concentration. In most cases, no significant difference was noted in the CIRs of both species when they were under the same light intensity and BOC. Nevertheless, it seems important to consider the impact of dissolved BOC during the treatment of bacterial polluted water.