The Gram-negative saprophyte Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease which is endemic in Southeast Asia and northern Australia. This bacterium possesses many virulence factors which are thought to contribute to its survival and pathogenicity. Using a virulent clinical isolate of B. pseudomallei and an attenuated strain of the same B. pseudomallei isolate, 6 genes BPSL2033, BP1026B_I2784, BP1026B_I2780, BURPS1106A_A0094, BURPS1106A_1131, and BURPS1710A_1419 were identified earlier by PCR-based subtractive hybridization. These genes were extensively characterized at the molecular level, together with an additional gene BPSL3147 that had been identified by other investigators. Through a reverse genetic approach, single-gene knockout mutants were successfully constructed by using site-specific insertion mutagenesis and were confirmed by PCR. BPSL2033::Km and BURPS1710A_1419::Km mutants showed reduced rates of survival inside macrophage RAW 264.7 cells and also low levels of virulence in the nematode infection model. BPSL2033::Km demonstrated weak statistical significance (P = 0.049) at 8 hours after infection in macrophage infection study but this was not seen in BURPS1710A_1419::Km. Nevertheless, complemented strains of both genes were able to partially restore the gene defects in both in vitro and in vivo studies, thus suggesting that they individually play a minor role in the virulence of B. pseudomallei.
Cannibalism is a major problem in lobster and crab aquaculture. Reducing the aggressive characteristics of lobsters and crabs can improve survival during the culturing process. In this study, juvenile scalloped spiny lobsters (Panulirus homarus) and crucifix crabs (Charybdis feriatus) were both cultured under different shelter and live prey conditions. Groups with shelter (seaweed and cotton filter) showed a better survival rate than the control group (no shelter; p < 0.05) for both Pa. homarus and Char. feriatus. Co-culturing with live prey (Litopenaeus vannamei) significantly benefited the juveniles of Pa. homarus and visibly increased the survival of juvenile Char. feriatus. Although providing shelter is currently the main method for reducing agonistic behavior, it must be continually altered as the lobsters and crabs grow. Live prey can grow and attract lobsters and crabs to hunt them, and live prey can be supplemented at any time. They can also be used as an additional source of income during the harvest season.