Application of polymerase chain reaction (PCR) and TaqMan PCR techniques to the detection and identification of Rhodococcus coprophilus in faecal samples.

Rhodococcus coprophilus, a natural inhabitant of herbivore faeces, has been suggested as a good indicator of animal (as opposed to human) faecal contamination of aquatic environments. However, conventional detection methods limit its use for this as they require up to 21 days to obtain a result. In this paper an optimised method for extracting R. coprophilus DNA from faecal samples is described. PCR and 5'-nuclease (TaqMan) PCR methods were developed to allow the detection and enumeration of R. coprophilus in faecal samples within 2-3 days. Both PCR methods targeted the 16S rRNA gene, producing an amplicon of 443 bp which was specific for R. coprophilus. Sixty cells were required to produce an amplification product by conventional PCR, while as little as one cell was required for the TaqMan PCR method. The latter approach gave a linear quantitative response over at least four log units with both bacterial cells and DNA. Successful amplification by PCR was achieved using DNA extracted from cow, sheep, horse and deer faeces but was negative for samples from humans, pig, possum, duck and rabbit. These PCR methods enhance the feasibility of using R. coprophilus to distinguish faecal pollution of farmed herbivores from human pollution.

A slim 198gold-grain implanter loaded with standard Royal Marsden 14-grain magazines.

We designed a slim gold-grain implanter with adaptable lengths to implant areas accessible only through long, narrow, examining instruments, such as a suspension laryngoscope. The implanter is loaded with the same 14-grain magazine designed for and supplied with the Royal Marsden "gun." The simplicity of the loading mechanism with a minimum of moving parts makes the instrument practically trouble free. Although it is designed to be used along narrow examining instruments, it can also be used in any situation in which a permanent implant is required, for instance, prostatic cancer and pelvic recurrences in cancer of the uterine cervix previously treated by external and intracavitary irradiation.