Malakar, P. K. | Search bioRxiv

bioRxiv2019

Cold Shock Fail to Restrain Pre-formed Vibrio parahaemolyticus Biofilm

The source of persistent infections can be biofilms that occur naturally on food surfaces and medical biomaterials. Biofilm formation on these materials are likely to be affected by environmental temperature fluctuations and information on noticeable temperature shifts on the fate of pre-formed biofilm is sparse. Changes to pre-formed Vibrio parahaemolyticus biofilm under cold shock (4 {degrees}C and 10 {degrees}C) was explored in this study. We show that V. parahaemolyticus biofilm biomass increased significantly during this cold shock period and there was a gradual increase of polysaccharides and proteins content in the extracellular polymeric matrix (EPS). In addition, we demonstrate that the expression of flagella and virulence-related genes were differentially regulated. The architecture of the biofilm, quantified using mean thickness (MT), average diffusion distance (ADD), porosity (P), biofilm roughness (BR) and homogeneity (H) also changed during the cold shock and these parameters were correlated (P < 0.01). However, the correlation between biofilm architecture and biofilm-related genes expression was relatively weak (P < 0.05). Cold shock at 4 {degrees}C and 10 {degrees}C is not sufficient to reduce V. parahaemolyticus biofilm formation and strategies to reduce risk of foodborne infections should take this information into account.

microbiology↗

bioRxiv2018

Cold Shock Fail to Restrain Pre-formed Bacterial Biofilm

Environmental temperature fluctuation has great impact on the formation of bacterial biofilm, while little information is available for assessing the influence of sharp temperature shifts on the fate of pre-formed biofilm. In this study, experimental evidence is firstly explored on the response of Vibrio parahaemolyticus pre-formed biofilm under cold shock (4 {degrees}C and 10 {degrees}C). Surprisingly, biofilm biomass of V. parahaemolyticus significantly increased during the period of cold shock as revealed by crystal violet staining. Polysaccharides and proteins contents in extracellular polymeric substances were gradually enhanced after cold shocks and exhibited high consistency. RT-qPCR demonstrated the expression of flagella and virulence-related genes were up-regulated. Most of QS and T3SS genes were slightly up-regulated, and three T3SS genes (vcrD1, vcrD2{beta} and vopD1) were down-regulated. Furthermore, the biofilm structure of V parahaemolyticus have been analyzed by Confocal laser scanning microscopy (CLSM), which sharply changed under cold shocks. The correlation analysis further displayed the significant correlation (P < 0.01) among biofilm structure parameters, and weak correlation (P < 0.05) between biofilm related genes and biofilm structure parameters. In conclusion, our results novel discovered that V. parahaemolyticus biofilm related genes were actively expressed and biofilm biomass was continuously increased, biofilm structure was tremendously changed after cold shock. This study underscored the risk that biofilm cells had the ability to adapt to low temperature shift.\n\nIMPORTANCEBiofilms are widespread in natural environments, especially on the surface of food and medical biomaterials, which threaten human safety from persistent infections. Previous studies simply focused on biofilm formation of microorganisms under steady state, however, the actual environment frequently fluctuated. V. parahaemolyticus is a widely distributed foodborne pathogen, temperature play a great role in its survival. Researchers generally assume that cold environment can restrain biofilm formation and bacterial activity. This study explored the effects of V. parahaemolyticus biofilm upon a shift from 37 {degrees}C to 4 {degrees}C or 10 {degrees}C from two aspects. On the one hand, the changes of biofilm biomass and EPS contents, the expression of biofilm related genes directly described that pre-formed bacterial biofilm could not be controlled efficiently in cold environment. On the other hand, the CLSM images revealed biofilm morphological structure change, the correlation analysis showed inner relationship among biofilm structure parameters and biofilm related genes. These results suggested that cold shock fail to restrain pre-formed bacterial biofilm, therefore be a potential risk in nature environment.

microbiology↗