Carbapenemases were rare at the time of this study. Dissemination of carbapenemases was due to both dominant clones and common plasmid backbones.

pPrY2001 differed from all known plasmids due to its novel backbone and repB. pKp11-42 was similar to IncFIIk plasmids and contained a number of genes that aid in plasmid persistence.

The bacterial spirochetes, Treponema spp., are thought to be a major contributor to the etiology of bovine digital dermatitis (DD), a skin disease with worldwide economic impact. Hoofbath strategies are commonly used in an attempt to control and prevent the development of DD and continuing research has been done to develop an optimal hoofbath strategy for this purpose. The aim of this study was to develop a protocol that can be used as part of the screening process for candidate hoofbath disinfectants. This protocol allows an accurate determination of the in vitro minimum inhibitory concentration and minimum bactericidal concentration of a series of disinfectants for Treponema microorganisms. Assays were performed in triplicate for each of the disinfectants at 30-s and 10-min exposure times and exposed to 10 and 20% manure (vol/vol). The results of this study can be used to categorize disinfectants based on the effect of exposure and manure concentration regarding their ability to inhibit Treponema growth. This information can then aid in optimizing strategies for hoofbath-based control of DD development and spread.

We present a method that allows preparing long DNA containing defined mismatches without the use of gel electrophoretic or chromatographic purification steps. The preparation starts with the synthesis of two PCR products, which are identical except for those positions that will later form the mismatches. One of the PCR primers must be 5'-phosphorylated, such that in two separate reactions two PCR products are obtained, which are 5'-phosphorylated in one strand. After removal of the phosphorylated strands by lambda-exonuclease, the resulting single strands are hybridized to form the mismatch-containing heteroduplex. The application of this procedure is demonstrated for the analysis of the Escherichia coli MutHLS system.

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