RESUMO
Growing concerns exist regarding human ingestion of contaminated seafood that contains Vibrio biofilms on microplastics (MPs). One of the mechanisms enhancing biofilm related infections in humans is due to biofilm dispersion, a process that triggers release of bacteria from biofilms into the surrounding environment, such as the gastrointestinal tract of human hosts. Dispersal of cells from biofilms can occur in response to environmental conditions such as sudden changes in temperature, pH and nutrient conditions, as the bacteria leave the biofilm to find a more stable environment to colonize. This study evaluated how brief exposures to nutrient starvation, elevated temperature, different pH levels and simulated human media affect Vibrio parahaemolyticus and Vibrio vulnificus biofilm dispersal and processes on and from low-density polyethylene (LDPE), polypropylene (PP), and polystyrene (PS) MPs. Both species were able to adequately disperse from all types of plastics under most exposure conditions. V. parahaemolyticus was able to tolerate and survive the low pH that resembles the gastric environment compared to V. vulnificus. pH had a significantly (p ≤ 0.05) positive effect on overall V. parahaemolyticus biofilm biomass in microplates and cell colonization from PP and PS. pH also had a positive effect on V. vulnificus cell colonization from LDPE and PP. However, most biofilm biomass, biofilm cell and dispersal cell densities of both species greatly varied after exposure to elevated temperature, pH, and nutrient starvation. It was also found that certain exposures to simulated human media affected both V. parahaemolyticus and V. vulnificus biofilm biomass and biofilm cell densities on LDPE, PP and PS compared to exposure to traditional media of similar pH. Cyclic-di-GMP was higher in biofilm cells compared to dispersal cells, but exposure to more stressful conditions significantly increased signal concentrations in both biofilm and dispersal states. Taken together, this study suggests that human pathogenic strains of V. parahaemolyticus and V. vulnificus can rapidly disperse with high cell densities from different plastic types in vitro. However, the biofilm dispersal process is highly variable, species specific and dependent on plastic type, especially under different human body related environmental exposures.
RESUMO
Vibrio vulnificus is an opportunistic pathogen indigenous to estuarine and marine environments and associated with aquatic organisms. Vibrio vulnificus is of utmost importance because it causes 95% of the seafood-related deaths in the United States due to rapid progression of septicemia. Changes in environmental parameters associated with climate change and coastal population expansion are altering geographical constraints, resulting in increased Vibrio spread, exposure, and rates of infection. In addition, coastal population expansion is resulting in increased input of treated municipal sewage into areas that are also experiencing increased Vibrio proliferation. This study aimed to better understand the influence of treated sewage effluent on effluent-receiving microbial communities using Vibrio as a model of an opportunistic pathogen. Integrated transcriptomic approaches were used to analyze the changes in overall gene expression of V. vulnificus NBRC 15645 exposed to wastewater treatment plant (WWTP) effluent for a period of 6h using a modified seawater yeast extract media that contained 0, 50, and 100% filtered WWTP effluent. RNA-seq reads were mapped, annotated, and analyzed to identify differentially expressed genes using the Pathosystems Resource Integration Center analysis tool. The study revealed that V. vulnificus responds to wastewater effluent exposure by activating cyclic-di-GMP-influenced biofilm development. Also, genes involved in crucial functions, such as nitrogen metabolism and bacterial attachment, were upregulated depending on the presence of treated municipal sewage. This altered gene expression increased V. vulnificus growth and proliferation and enhanced genes and pathways involved in bacterial survival during the early stages of infection in a host. These factors represent a potential public health risk due to exposure to environmental reservoirs of potentially Vibrio strains with enhanced virulence profiles in coastal areas.
RESUMO
Microspatial arrangements of sulfate-reducing microorganisms (SRM) in surface microbial mats (~1.5 mm) forming open marine stromatolites were investigated. Previous research revealed three different mat types associated with these stromatolites, each with a unique petrographic signature. Here we focused on comparing "non-lithifying" (Type-1) and "lithifying" (Type-2) mats. Our results revealed three major trends: (1) Molecular typing using the dsrA probe revealed a shift in the SRM community composition between Type-1 and Type-2 mats. Fluorescence in-situ hybridization (FISH) coupled to confocal scanning-laser microscopy (CSLM)-based image analyses, and 35SO4(2-)-silver foil patterns showed that SRM were present in surfaces of both mat types, but in significantly (p < 0.05) higher abundances in Type-2 mats. Over 85% of SRM cells in the top 0.5 mm of Type-2 mats were contained in a dense 130 µm thick horizontal layer comprised of clusters of varying sizes; (2) Microspatial mapping revealed that locations of SRM and CaCO3 precipitation were significantly correlated (p < 0.05); (3) Extracts from Type-2 mats contained acylhomoserine-lactones (C4- ,C6- ,oxo-C6,C7- ,C8- ,C10- ,C12- , C14-AHLs) involved in cell-cell communication. Similar AHLs were produced by SRM mat-isolates. These trends suggest that development of a microspatially-organized SRM community is closely-associated with the hallmark transition of stromatolite surface mats from a non-lithifying to a lithifying state.
