Modification of Medium Composition for Enhancing the Production of Antifungal Activity from Xenorhabdus stockiae PB09 by Using Response Surface Methodology.

Xenorhabdus stockiae PB09 bacterium has been shown to exhibit antifungal activity against several plant pathogens. To improve its efficacy, the optimization of the nutritional components in culture media was performed. The medium components that have significant effects on antifungal activity of X. stockiae PB09 were initially identified using a fractional factorial design. Response surface methodology and central composite design were then used to create a model for optimizing the levels of carbon, nitrogen, and mineral sources that maximize antifungal activity of X. stockiae PB09. After that, the suitable carbon, nitrogen, and mineral sources were selected and adjusted by the second-order polynomial regression model, which predicted that 98.62% of antifungal activity could be obtained when the medium contained sucrose, yeast extract, NaCl, and K2HPO4 at 3.24, 23.71, 5.46, and 2.73 g/L, respectively. Laboratory verification of this recipe resulted in the antifungal activity at 97.95% in the shake flask experiment after 48-hour cultivation, which was significantly 27.22% higher than that obtained by using the TSB medium. In addition, X. stockiae PB09 cultured in the verified recipe by using 5 L fermenter could effectively inhibit the mycelial growth of Phytophthora sp., Rhizoctonia solani, Pythium sp., and Fusarium oxysporum. This study demonstrated that the RSM and CCD were shown to be valuable tools for optimizing the culture medium that maximize the antifungal activity of X. stockiae PB09.

Fast Population Growth in Physogastry Reproduction of Luciaphorus perniciosus (Acari: Pygmephoridae) at Different Temperatures.

Luciaphorus perniciosus Rack is one of the most serious pests of several cultivated mushroom species including Ganoderma lucidum (Fr.), Flammulina velutipes Karst., Auricularia polytricha (Mont.) Saac., Lentinus polychrous Lev., and Lentinus squarrosulus (Mont.) Singer in Thailand. Adult female Lu. perniciosus produce offspring inside their physogastric hysterosomas, with all embryos developing through to the adult stage while remaining in the abdomen. Once the abdomen ruptures, the female parent dies and the offspring consisting of mostly fertilized female adults along with a few male adults continue to emerge from the cadaver of the mother for a period of several days. This peculiar type of reproduction after the death of the mother is a special case for life table analysis and has not been discussed previously in demographic analyses. In this study, the life table data of this mite fed on Le. squarrosulus were collected at 25, 30, and 35 °C and analyzed by using the age-stage, two-sex life table. The standard errors of population parameters were estimated by using the bootstrap technique (200,000 bootstraps). At 25, 30, and 35 °C, females started reproduction at ages 9, 5, and 3 d, respectively; the net reproductive rates (R0) were 192.27, 253.81, and 234.11 offspring. Due to their rapid development and high fecundity, the r values were as high as 0.4189, 0.8653, and 1.0892 d-1 at 25, 30, and 35 °C, respectively. Computer projection indicated that the mushroom mites Lu. perniciosus is capable of a threefold daily increase at 35 °C.

Acaricidal activities of whole cell suspension, cell-free supernatant, and crude cell extract of Xenorhabdus stokiae against mushroom mite (Luciaphorus sp.).

Xenorhabdus bacterium has been used as a biological control agent against Luciaphorus sp., a mushroom mite endemic in Thailand. To develop an effective formulation of Xenorhabdus stokiae, treatments using different parts of X. stokiae isolate PB09 culture, including whole cell suspension, cell-free supernatant, and crude cell extract, were performed. The results show that different parts of X. stokiae isolate PB09 culture could induce variable effects on mite mortality and fecundity. Application with cell-free supernatant of X. stokiae culture resulted in both the highest mite mortality rate [(89.00±3.60)%] and the lowest mite fecundity [(41.33±23.69) eggs/gravid female]. Whole cell suspension of X. stokiae isolate PB09 culture was found to be slightly less effective than its cell-free supernatant, suggesting that X. stokiae was more likely to release its metabolites with acaricidal activities to the surrounding culture media. Crude cell extract of X. stokiae was not effective against mites. Cell-free supernatant of X. stokiae isolate PB09 was the most effective biological control agent and it could be conveniently used in future formulations instead of live bacteria.