Quantitative analysis of anthrax spores from environmental samples is essential for accurate detection and risk assessment since Bacillus anthracis spores have been shown to be one of the most effective biological weapons. Using TaqMan real-time PCR, specific primers and probes were designed for the identification of pathogenic B. anthracis strains from pag gene and cap gene on two plasmids, pXO1 and pXO2, as well as a sap gene encoded on the S-layer. To select the appropriate lysis method of anthrax spore from environmental samples, several heat treatments and germination methods were evaluated with multiplex-PCR. Among them, heat treatment of samples suspended with sucrose plus non-ionic detergent was considered an effective spore disruption method because it detected up to 10 5 spores/g soil by multiplex-PCR. Serial dilutions of B. anthracis DNA and spore were detected up to a level of 0.1 ng/l and 10 spores/ml, respectively, at the correlation coefficient of 0.99 by real-time PCR. Quantitative analysis of anthrax spore could be obtained from the comparison between CT value and serial dilutions of soil sample at the correlation coefficient of 0.99. Additionally, spores added to soil samples were detected up to 10 4 spores/g soil within 3 hr by real-time PCR. As a consequence, we established a rapid and accurate detection system for environmental anthrax spores using real-time PCR, avoiding time and labor-consuming preparation steps such as enrichment culturing and DNA preparation.

This outbreak points to the importance of drinking water quality management in group facilities where underground water is used and emphasizes the need for periodic sanitation and inspection to prevent possible waterborne outbreaks.

No abstract

Antimicrobial-resistant gonococcus has been a major problem in sexually transmitted disease control. Outer membrane proteins (OMPs) of Neisseria gonorrhoeae were suggested to have influence on its resistance to antibiotics. So, in this work, we provide a proteomic analysis tool for examining the OMPs of N. gonorrhoeae and also provide a comparative analysis of the OMPs between the susceptible parent strain (92WT) and the resistance-induced isogenic mutant (92mu13) to determine the OMPs responsible for resistance. The 2-D gel spots of 92mu13 differed from 92WT particularly in porin, pilus secretion protein (PilQ) and enzymes. PilQ expression in 92mu13 was considerably reduced by abrupt termination at nucleotide 2,112. This made it difficult to form a high molecular mass (HMM) pore at the outer membrane; it is suspected that reduction of PilQ serves a role in antibiotic resistance in N. gonorrhoeae. The amount of porin was not changed but its isoelectric point (pI) shifted to a basic region, which is caused by the alteration of an amino acid of porin and it is suggested to relate to the development of antimicrobial resistance. Differential regulation of the enzymes involved in metabolism was found in 92mu13, believed to represent an adaptation of N. gonorrhoeae to the antibiotic environment.