DLVO, hydrophobic, capillary and hydrodynamic forces acting on bacteria at solid-air-water interfaces: Their relative impact on bacteria deposition mechanisms in unsaturated porous media.

Experimental and modeling studies were performed to investigate bacteria deposition behavior in unsaturated porous media. The coupled effect of different forces, acting on bacteria at solid-air-water interfaces and their relative importance on bacteria deposition mechanisms was explored by calculating Derjaguin-Landau-Verwey-Overbeek (DLVO) and non-DLVO interactions such as hydrophobic, capillary and hydrodynamic forces. Negatively charged non-motile bacteria and quartz sands were used in packed column experiments. The breakthrough curves and retention profiles of bacteria were simulated using the modified Mobile-IMmobile (MIM) model, to identify physico-chemical attachment or physical straining mechanisms involved in bacteria retention. These results indicated that both mechanisms might occur in both sand. However, the attachment was found to be a reversible process, because attachment coefficients were similar to those of detachment. DLVO calculations supported these results: the primary minimum did not exist, suggesting no permanent retention of bacteria to solid-water and air-water interfaces. Calculated hydrodynamic and resisting torques predicted that bacteria detachment in the secondary minimum might occur. The capillary potential energy was greater than DLVO, hydrophobic and hydrodynamic potential energies, suggesting that film straining by capillary forces might largely govern bacteria deposition under unsaturated conditions.

Bacteria cell properties and grain size impact on bacteria transport and deposition in porous media.

The simultaneous role of bacteria cell properties and porous media grain size on bacteria transport and deposition behavior was investigated in this study. Transport column experiments and numerical HYDRUS-1D simulations of three bacteria with different cell properties (Escherichia coli, Klebsiella oxytoca, and Rhodococcus rhodochrous) were carried out on two sandy media with different grain sizes, under saturated steady state flow conditions. Each bacterium was characterized by cell size and shape, cell motility, electrophoretic mobility, zeta potential, hydrophobicity and potential of interaction with the sand surface. Cell characteristics affected bacteria transport behavior in the fine sand, but similar bacteria breakthroughs and retardation factors observed in the coarse sand, indicated that bacteria transport was more depended on grain size than on bacteria cell properties. Retention decreased with increasing hydrophobicity and increased with increasing electrophoretic mobility of bacteria for both sand. The increasing sand grain size resulted in a decrease of bacteria retention, except for the motile E. coli, indicating that retention of this strain was more dependent on cell motility than on the sand grain size. Bacteria deposition coefficients obtained from numerical simulations of the retention profiles indicated that straining was an important mechanism affecting bacteria deposition of E. coli and Klebsiella sp., in the fine sand, but the attachment had the same importance as straining for R. rhodochrous. The results obtained in the coarse sand did not permit to discriminate the predominant mechanism of bacteria deposition and the relative implication of bacteria cell properties of this process.

Passaging impact of H9N2 avian influenza virus in hamsters on its pathogenicity and genetic variability.

INTRODUCTION: Avian influenza viruses of the H9N2 subtype have been reported to cause human infections. This study demonstrates the impact of nasal viral passaging of avian H9N2 in hamsters on its cross species-pathogenic adaptability and variability of amino acid sequences of the hemagglutinin (HA) and neuraminidase (NA) stalk. METHODOLOGY: Three intranasal passagings of avian H9N2 in hamsters P1, P2, and P3 were accomplished. Morbidity signs and lesions were observed three days post viral inoculation. The HA test was used for presumptive detection of H9N2 virus in the trachea and lungs of the hamsters challenged with the differently passaged viruses. Different primers were used for PCR amplification of the HA1 and NA stalk regions of the differently passaged H9N2 viruses, followed by sequence alignment. RESULTS: The morbidity signs indicated low pathogenicity of the differently passaged H9N2 viruses in hamsters. The frequency of gross and microscopic lesions in the tracheas and lungs were insignificantly different among hamsters challenged with the differently passaged H9N2 viruses (p > 0.05). There was 100% similarity in the amino acid sequence of the HA gene of most passaged viruses. The amino acid sequence of the neuraminidase in the third passaged H9N2 virus recovered from lungs showed a R46P mutation that might have a role in the pathogenic adaptability of P3 viruses in hamsters' lungs. CONCLUSIONS: The apparent adaptation of avian H9N2 virus to mammalian cells is in agreement with the World Health Organization's alertness for a possible public health threat by this adaptable virus.

Impact of embryonic passaging of H9N2 virus on pathogenicity and stability of HA1- amino acid sequence cleavage site.

BACKGROUND: Highly virulent Avian Influenza viruses might arise from avirulent strains following viral passaging. This work aims at studying the impact of embryonic passaging of H9N2 on the stability of the HA1 amino acid sequence and its relatedness to pathogenicity. MATERIAL/METHODS: The original H9N2 virus was propagated for 3 consecutive passages in embryonated chicken eggs. Pathogenicity and amino acids sequences at the HA1 gene level of the original (P0), and the once (P1), twice (P2), and three times (P3) passaged viruses were compared. RESULTS: The percent mortality significantly increased in embryos inoculated with P2 (86.7%) and P3 of H9N2 (100%) in comparison to P0 (0.0%) and P1 of H9N2 (46.1%) (P<0.05), while the density of propagated H9N2 declined with passaging. The R-S-S-R motif was stable at the HA1 cleavage site of P0, P1, P2, and P3 viruses. The similarity in the HA1 sequences among the differently passaged viruses ranged between 93.2 to 100%. CONCLUSIONS: The pathogenicity increased significantly upon passaging in chicken embryos in spite of the presence of the same motif at the HA1 cleavage site. Further investigations will target the study of changes in the whole HA protein and of Neuraminidases that could be responsible for a higher pathogenicity.

Monitoring the impact of hydrocarbon contamination and nutrient addition on microbial density, activity, and diversity in soil.

The development of optimal in situ bioremediation strategies requires a better knowledge of their impact on the soil microbial communities. We have evaluated the impact of hexadecane contamination and different nutrient amendments on soil microbial density and activity. Microbial density was measured via total DNA quantification, and microbial activity via respiration and RNA variation. The RNA/DNA ratio was also determined, as it is a potential indicator of microbial activity. PCR-amplified 16S rRNA genes were cloned and sequenced to analyze the diversity of bacterial communities. Nutrient addition significantly increased respiration and DNA and RNA concentrations in contaminated soil, indicating a limitation of degradation and growth by the availability of nitrogen and phosphorus in unamended microcosms. Hexadecane treatment slightly affected the diversity of the bacterial community, while it was dramatically reduced by nutrient treatments, particularly the addition of nitrogen and phosphorus. Microbial community composition was also altered with the enrichment of populations related to Nocardia in bioremediated soils, while uncultured Proteobacteria were mostly detected in uncontaminated soil.