Detection of Multiple Resistances, Biofilm Formation and Conjugative Transfer of Bacillus cereus from Contaminated Soils.

The purpose of this study was to detect microbial resistances to a set of antibiotics/pesticides (multi-resistance) within pesticide and antibiotic-contaminated alluvial soils and to identify the corresponding antibiotic resistance genes (ARGs). To assess whether identified multi-resistant isolates are able to construct biofilms, several biofilm formation and conjugation experiments were conducted. Out of 35 isolates, six strains were used for filter mating experiments. Nine strains were identified by 16S rDNA gene sequence analyses and those were closely related to Pseudomonas sp., Citrobacter sp., Acinetobacter sp., Enterobacter sp., and in addition, Bacillus cereus was chosen for multi-resistant and pesticide-tolerant studies. Antibiotic-resistant and pesticide-tolerant bacterial strains were tested for the presence of ARGs. All nine strains were containing multiple ARGs (ampC, ermB, ermD, ermG, mecA, tetM) in different combinations. Interestingly, only strain WR34 (strongly related to Bacillus cereus) exhibited a high biofilm forming capacity on glass beads. Results obtained by filter mating experiments demonstrated gene transfer frequencies from 10(-5) to 10(-8). This study provides evidence that alluvial soils are hot spots for the accumulation of antibiotics, pesticides and biofilm formation. Particularly high resistances to tetracycline, ampicillin, amoxicillin and methicillin were proved. Apparently, isolate WR34 strongly correlated to a pathogenic organism had high potential to deploy biofilms in alluvial soils. Thus, we assume that loosened and unconsolidated soils investigated pose a high risk of an enhanced ARG prevalence.