Elucidating the genetic basis of crystalline biofilm formation in Proteus mirabilis.

Proteus mirabilis forms extensive crystalline biofilms on urethral catheters that occlude urine flow and frequently complicate the management of long-term-catheterized patients. Here, using random transposon mutagenesis in conjunction with in vitro models of the catheterized urinary tract, we elucidate the mechanisms underpinning the formation of crystalline biofilms by P. mirabilis. Mutants identified as defective in blockage of urethral catheters had disruptions in genes involved in nitrogen metabolism and efflux systems but were unaffected in general growth, survival in bladder model systems, or the ability to elevate urinary pH. Imaging of biofilms directly on catheter surfaces, along with quantification of levels of encrustation and biomass, confirmed that the mutants were attenuated specifically in the ability to form crystalline biofilms compared with that of the wild type. However, the biofilm-deficient phenotype of these mutants was not due to deficiencies in attachment to catheter biomaterials, and defects in later stages of biofilm development were indicated. For one blocking-deficient mutant, the disrupted gene (encoding a putative multidrug efflux pump) was also found to be associated with susceptibility to fosfomycin, and loss of this system or general inhibition of efflux pumps increased sensitivity to this antibiotic. Furthermore, homologues of this system were found to be widely distributed among other common pathogens of the catheterized urinary tract. Overall, our findings provide fundamental new insight into crystalline biofilm formation by P. mirabilis, including the link between biofilm formation and antibiotic resistance in this organism, and indicate a potential role for efflux pump inhibitors in the treatment or prevention of P. mirabilis crystalline biofilms.

Occurrence of N-nitrosodimethylamine precursors in wastewater treatment plant effluent and their fate during ultrafiltration-reverse osmosis membrane treatment.

The formation of N-nitrosodimethylamine (NDMA) is of major concern among wastewater recycling utilities practicing disinfection with chloramines. The NDMA formation potential (FP) test is a simple and straightforward method to evaluate NDMA precursor concentrations in waters. In this paper we show the NDMA FP results of a range of tertiary wastewater treatment plants that are also the source for production of recycled water using an Ultrafiltration - Reverse Osmosis (UF-RO) membrane process. The results indicate that the NDMA FP of different source waters range from 350 to 1020±20 ng/L. The fate of these NDMA precursors was also studied across the different stages of two Advanced Water Treatment Plants (AWTP) producing recycled water. These results show that more than 98.5±0.5% of NDMA precursors are effectively removed by the Reverse Osmosis (RO) membranes used at the AWTPs. This drastically reduces any potential for re-formation of NDMA after the RO stage even if chloramines may be present (or added) there.

Quantitative GC-MS analysis of D(9)-tetrahydrocannabinol in fiber hemp varieties.

A sensitive and specific method for the determination of total available D(9)-tetrahydrocannabinol in fiber hemp varieties is described. The method was used for the regulatory purposes in which the detection of higher than the maximum allowed concentration of the psychoactive cannabinoid, D(9)-tetrahydrocannabinol, in industrial fiber hemp would result in cancellation of the grower's license. Cannabinoids were extracted from dry leaf powder into hexane containing internal standard chrysene-d(12) using sonication. D(9)-Tetrahydrocannabinol in the extract was separated by gas chromatography and quantitated by mass spectroscopy. A linear calibration range extending to 40 ppm and a limit of detection of 0.2 ng were obtained by using the total ion current mode of detection.