Melioidosis was first recognized in northeastern Brazil in 2003. Confirmation of additional cases from the 2003 cluster in Ceará, more recent cases in other districts, environmental isolation of Burkholderia pseudomallei, molecular confirmation and typing results, and positive serosurveillance specimens indicate that melioidosis is more widespread in northeastern Brazil than previously thought.

Isolation of Burkholderia pseudomallei currently relies on the use of Ashdown's selective agar (ASA). We designed a new selective agar (Burkholderia pseudomallei selective agar [BPSA]) to improve recovery of the more easily inhibited strains of B. pseudomallei. B. pseudomallei, Burkholderia cepacia, and Pseudomonas aeruginosa were used to determine the selectivity and sensitivity of BPSA. BPSA was more inhibitory to P. aeruginosa and B. cepacia and should make recognition of Burkholderia species easier due to distinctive colony morphology. BPSA also inhibited Enterococcus, Escherichia, Staphylococcus, and Streptococcus. These results indicate that BPSA is a potential replacement for ASA.

The effect of chlorine, monochloramine and UV disinfection on the water-borne pathogen Burkholderia pseudomallei was assessed. Persistence of B. pseudomallei was verified by MPN involving a one-step recovery procedure. Chlorine proved the most effective disinfectant with a 99.99% reduction of a 10 6 CFU/mL pure bacterial culture followed by 99.9% reduction by monochloramine and 99% reduction by UV. Co-culture of B. pseudomallei with Acanthamoeba astronyxis was found to greatly enhance survival of B. pseudomallei in the presence of all disinfecting agents tested. For example, when amoebae were present 100 times more monochloramine was required to maintain the disinfectant efficacy. Given the results obtained from these co-culture experiments, more research is needed to investigate the role of amoeba and biofilms in survival of B. pseudomallei in potable water.

Chlorine is widely used in public water supplies to provide a disinfection barrier. The effect of chlorine disinfection on the water-borne pathogen Burkholderia pseudomallei was assessed using multiple techniques. After exposure to chlorine viable bacteria were undetectable by conventional plate count techniques, however persistence of B. pseudomallei was verified by flow cytometry and bacteria were recoverable following a simple one step broth procedure. The minimum residual chlorine concentration and contact time as prescribed by potable water providers in Australia was insufficient to reduce a B. pseudomallei population by more than 2 log 10 . Chlorine had a bacteriostatic effect only on B. pseudomallei; viable bacteria were recovered from water containing up to 1000 ppm free chlorine. This finding has practical implications for water treatment in regions where B. pseudomallei is endemic. Future work to assess the effect of alternative water disinfection processes either singly or in sequence is necessary.

Melioidosis was first recognized in northeastern Brazil in 2003. Confirmation of additional cases from the 2003 cluster in Ceará, more recent cases in other districts, environmental isolation of Burkholderia pseudomallei, molecular confirmation and typing results, and positive serosurveillance specimens indicate that melioidosis is more widespread in northeastern Brazil than previously thought.