Impact of metal exposure on environmentally isolated Serratia marcescens' growth, oxidative-stress resistance, biofilm formation, and proliferation in eukaryotic co-culture models.

Environmental metals can be noxious to the surrounding biota, indirectly impact freshwater habitats, and also impact microbiological communities. In this study, zinc (Zn) (55.5 mg/kg), manganese (Mn) (863.4 mg/kg) and lead (Pb) (17.5 mg/kg) levels measured in Houston watershed flood plain soil samples were higher than environmental agencies' thresholds. To investigate the effects of metal exposures, an environmentally isolated Serratia marcescens (SME), etiological agent of endocarditis and respiratory infections, and its reference strain (SMR) were exposed to Pb, Zn, and Mn, and subsequent oxidative stress responses and biofilm production were measured. Not surprisingly, SME was less sensitive to all 3 metal exposures than was SMR. Interestingly, SME produced increased biofilm and was more resistant to oxidative stress in the presence of Zn and Pb than SMR. In a 6 h lung infection model using BAES-2B cells, SME exhibited greater proliferation than SMR in all metal challenges. Similarly, in our HT29 gut infection model, SME out-proliferated SMR when challenged with Pb and Mn following the 6 h infection. Taken together, SME was better able to withstand environmental stressors than SMR, suggesting increased virulence potential of this opportunistic human pathogen.

Cytotoxicity analysis of pre- and post-hurricane harvey soil samples collected from greater houston bayous.

Rapid urbanization, anthropogenic pollution and frequent flooding events are affecting the soil and water quality along the streams and bayous of Houston. Soil acts as sink and reservoir of heavy metals and nutrients affecting human and animal health. The objectives of the study are 1) to analyze the effects of the metal and nutrient concentration of bayou flood plain surface soil samples on the gut cell cytotoxicity and 2) to evaluate the spatial and temporal difference in soil contamination on cell viability of colon cancer (HT-29) and normal colon epithelial (CCD 841 CoN) cell lines. To evaluate soil contamination between pre- and post-hurricane (Summer and Fall) conditions in six Bayous (Brays, Buffalo, Halls, Hunting, Greens and White Oak Bayous) of Harris County, Texas, in vitro bioassay analysis was applied to soil extracts. The MTT assay determined that, with increase in concentration of Bayou soil from 12.5% to 100%, the viability of CCD 841 CoN and HT-29 cells decreased significantly, across all sampling locations during both summer and fall seasons. Among all the bayous, the viability of CCD 841 CoN cells in summer and fall followed the pattern of White Oak > Greens > Halls > Brays Bayou, where the viability of cells exposed to White Oak soils was 3-4 times higher than cells exposed to Brays Bayou soil at 100% soil concentration. The viability of HT-29 cells in both seasons followed the pattern of Greens > White Oak > Halls > Brays Bayou, where the viability of cells exposed to Greens Bayou soil was more than 3-4 times higher than the cells exposed to Brays Bayou soil at 100% concentration. The higher concentration of metals and nutrients such as P, Zn, Cd, and Cu might have contributed to the significant cell lethality in Brays Bayou samples compared to other locations.

Klebsiella spp. isolates from Houston bayous exhibit increased resistance to lead exposure and possess enhanced virulence potential.

Houston watersheds are susceptible to microbial contamination as well as chemical contaminations from bordering industrial facilities. Bacterial loads in various Houston bayous were determined, and pathogenic Gram-negative bacteria were isolated for characterization. Isolates included Klebsiella aerogenes and Klebsiella pneumoniae. To determine whether environmental exposures to lead (Pb), measured in our Houston bayou samples, resulted in bacterial adaptations, we compared growth kinetics, biofilm production, oxidative stress resistance, and eukaryotic co-culture growth of environmentally isolated K. aerogenes and K. pneumoniae to their respective commercially acquired reference strains. Interestingly, the K. aerogenes environmental isolate displayed significantly better growth than the reference strain in the presence of 50 ppb of Pb. Unexpectedly, we did not observe any differences in biofilm production of the aforementioned strains when challenged with a range of Pb (0.5-50 ppb). However, when comparing our K. pneumoniae environmental isolate to its reference strain, there were significantly higher levels of biofilm produced by the environmental isolate when challenged with Pb concentrations of 10 and 50 ppb. When grown in eukaryotic cell co-culture with either BAES 2B lung cells or CCD 841 colon epithelial cells in the presence of 20 ppb Pb, the environmental isolates of K. aerogenes and K. pneumoniae had a significantly higher fold-increase over 6 h than their respective reference strains. Taken together, the environmentally isolated Klebsiella spp. appeared to be more Pb-tolerant than their respective reference strains, a possible environmental adaptation. Such enhanced tolerance can promote environmental persistence and increase the possibility of causing human disease.