Isolation and identification of Burkholderia pseudomallei from soil using selective culture techniques and the polymerase chain reaction.

An environmental soil survey to detect Burkholderia pseudomallei was performed during the dry and wet seasons in Darwin, Northern Territory, Australia. Soil was sampled at regular intervals during a 15-month period at different depths from areas which were representative of the local, soil environment. Selective culture techniques using Ashdown's and Galimand and Dodin's methods and the polymerase chain reaction (PCR) using specific 16S rRNA primers were used to detect and identify the organism and determine its distribution within the soil stratum over the change in seasons. Results showed that Ashdown's method gave higher isolation rates in the dry season, and Galimand and Dodin's method gave higher isolation rates during the wet season. PCR of the soil enrichment proved to be a more sensitive method than culture and was also a useful confirmatory test in determining the identification of isolates where biochemical tests gave inconsistent results. The PCR primers were specific and able to detect 10(1) cfu g-1 soil and 10(4) cfu g-1 of soil using Ashdown's enrichment broth and Galimand and Dodin's broth, respectively. Overall the isolation of B. pseudomallei was greatest during the dry season and at the higher and lower soil depths, which is contradictory to epidemiological evidence that melioidosis occurs primarily during the wet season among patients exposed to contaminated surface soil and water.

Sensitivity of Australian isolates of Salmonella enteritidis to nitrofurantoin and furazolidone.

Susceptibility of 66 and 62 Australian isolates of Salmonella enteritidis to nitrofurantoin and furazolidone, respectively, was determined. Most isolates were susceptible to both antibiotics. Cross-resistance was low among all isolates, but higher among the subset of phage type 4 isolates. These results contrast directly with those of a previous study (Rampling, A., Upson, R. and Brown, D.F.J. (1990) J. Antimicrob. Chemother., 25: 285-290). Sensitivity among Australian isolates of S. enteritidis does to some extent, support the contention that furans may have played a role in the selection and enhanced colonisation of poultry by Salmonella enteritidis in Britain. Furthermore, nitrofurantoin should not be used as a selective agent in the isolation of Salmonella enteritidis.