Effects of Heavy Metal Stress on the Growth Curves of Two Sepcies of Bacteria

[Objective] The aim was to study on the effects of heavy metal stress on the growth curves of Escherichia coli and Bacillus subtili. [Method] Using traditional culture method,Escherichia coli and Bacillus subtili were cultured under heavy metal stress including Cu2＋,Hg2＋,Pb2＋,Cd2＋ and Cr6＋ in different concentrations. Then the growth curves of the bacteria were determined to investigate the effects of exogenous heavy metals on the growth of the two kinds of bacteria. [Result] The proliferation of the two bacteria was inhibited at high concentrations of Hg2＋ and Cd2＋ respectively,and G＋ is more sensitive to them than G-; when the heavy metal concentration was 50 mg/L,the toxicity of the five kinds of heavy metals on the two bacteria was Hg2＋Cd2＋Cu2＋Cr6＋≈Pb2＋. [Conclusion] The research will provide a basis to explore the effects of heavy metal on environment and ecological system.

Effect of Bacillus baekryungensis YD13 Supplemented in Diets on Growth Performance and Immune Response of Sea Cucumber(Apostichopus japonicus)

The effect of a potential probiotic on the growth performance and immune response of sea cucumber(Apostichopus japonicus) was investigated. Bacillus baekryungensis YD13 isolated from sea cucumber culturing ponds was added to sea cucumber basal feed as a probiotic in different doses(0, the control; 1×104(YD134), 1×106(YD136) and 1×108(YD138) CFU g-1 of diet), and administered orally to A. japonicus(initial mean wet weight 5.44 g ± 0.17 g). The sea cucumbers were fed in 20 aquaria, 5 each treatment, for 60 d. At the end of growth trial, 20 sea cucumbers from each treatment were challenged with Vibrio splendidus. A. japonicus in YD134 and YD136 exhibited significantly better growth performance than control(P < 0.05). Five non-specific immune parameters including lysozyme, acid phosphatase, alkaline phosphatase, superoxide dismutase and catalase in coelomic fluid were measured to evaluate the immune response of A. japonicus to the probiotics. Results showed that all parameters were significantly improved when YD11 was supplemented in the dose of 1×106 CFU g-1(P < 0.05). The cumulative incidence and mortality after the Vibrio splendidus challenge decreased significantly in sea cucumbers of YD136. Accordingly, 1×106 CFU g-1 of YD13 in diet was recommended for the growth promotion and immune enhancement of A. japonicus.

Antifouling Potential of Bacteria Isolated from a Marine Biofilm

Marine microorganisms are a new source of natural antifouling compounds. In this study, two bacterial strains, Kytococcus sedentarius QDG-B506 and Bacillus cereus QDG-B509, were isolated from a marine biofilm and identified. The bacteria fermentation broth could exert inhibitory effects on the growth of Skeletonema costatum and barnacle larvae. A procedure was employed to extract and identify the antifouling compounds. Firstly, a toxicity test was conducted by graduated pH and liquid-liquid extraction to determine the optimal extraction conditions. The best extraction conditions were found to be pH 2 and 100% petroleum ether. The EC50 value of the crude extract of K. sedentarius against the test microalgae was 236.7 ± 14.08 μg mL-1, and that of B. cereus was 290.6 ± 27.11 μg mL-1. Secondly, HLB SPE columns were used to purify the two crude extracts. After purification, the antifouling activities of the two extracts significantly increased: the EC50 of the K. sedentarius extract against the test microalgae was 86.4 ± 3.71 μg mL-1, and that of B. cereus was 92.6 ± 1.47 μg mL-1. These results suggest that the metabolites produced by the two bacterial strains are with high antifouling activities and they should be fatty acid compounds. Lastly, GC-MS was used for the structural elucidation of the compounds. The results show that the antifouling compounds produced by the two bacterial strains are myristic, palmitic and octadecanoic acids.

Isolation of Six Phosphate Dissolving Rhizosphere Bacteria （Bacillus subtilis） and Their Effects on the Growth, Phosphorus Nutrition and Yield of Maize （Zea mays L.） in Mali

The disadvantages of the long term application of Tilemsi natural phosphate （TNP） on maize （Zea mays L.） production has come out because of its low P availability. Some functional soil microbes, such as phosphate dissolving bacteria, have great potential in improvement of P solubility from TNP and P uptake by plants. The present study aimed to isolate and characterize typical phosphate dissolving bacterial strains （Bacillus subtilis） from Malian soils, and investigate their role in P uptake by maize grown in soils amended with TNP. The experimental design was a split plot with three main plots of fertilizers sources, i.e., natural phosphate, commercial fertilizer and without fertilizer, and with seven sub-plots of six microorganisms plus the control. The field experiment results have shown that the maize inoculated with the phosphate dissolving bacteria was improved in seed germination, plant growth, plant production （increase yield by 42%）, grain and aerial dry biomass （P） content of 34% and 64%, respectively. They have also shown that the locally available TNP can be used by the Malians farmers in maize culture and have comparable production to the one obtained with the costly imported commercial phosphate fertilizer, like the complex cereal. The project has provided information for the combined use of the Mali TNP and phosphate dissolving bacteria Bacillus subtilis subsp, subtilis （T）： DSM 10 in improvement of maize production in the country.

Bacillus promotes invasiveness of exotic Flaveria bidentis by increasing its nitrogen and phosphorus uptake

The effect of exotic plants on Bacillus diversity in the rhizosphere and the role of Bacilli in exotic or native plant species remain poorly understood.Flaveria bidentis is an invasive grass in China.Setaria viridis is a native grass and occurs in areas invaded by F.bidentis.Our objectives were(i)to examine the differences in the Bacillus communities between F.bidentis and S.viridis rhizospheres soil,and(ii)to compare the effects of Bacilli from F.bidentis and S.viridis rhizospheres on the competitiveness of the invasive species.Flaveria bidentis monoculture,mixture of F.bidentis and S.viridis and S.viridis monoculture were designed in the field experiment.Bacillus diversity in their rhizosphere was analyzed using 16S rRNA.One of the dominant Bacilli in the rhizosphere soil of F.bidentis was selected to test its effect on the competitive growth of F.bidentis in a greenhouse experiment.Bacillus diversity differed in F.bidentis and S.viridis rhizosphere.Brevibacterium frigoritolerans was the dominant Bacilli in the rhizosphere of both F.bidentis and S.viridis;however,its relative abundance in the F.bidentis rhizosphere was much higher than that in the S.viridis rhizosphere.In addition,B.frigoritolerans in the F.bidentis rhizosphere enhanced the growth of the plant compared with that of S.viridis by improving the nitrogen and phosphorus levels.This study showed that F.bidentis invasion influenced Bacillus communities,especially B.frigoritolerans,which,in turn,facilitated F.bidentis growth by increasing the levels of available nitrogen and phosphorus.

Bacillus amyloliquefaciens Strain W19 can Promote Growth and Yield and Suppress Fusarium Wilt in Banana Under Greenhouse and Field Conditions

Plant growth-promoting rhizobacteria （PGPR） are considered to be the most promising agents for cash crop production via increasing crop yields and decreasing disease occurrence. The Bacillus amyloliquefaciens strain W19 can produce secondary metabolites （iturin and bacillomycin D） effectively against Fusarium oxysporum f. sp. cubense （FOC）. In this study, the ability of a bio-organic fertilizer （BIO） containing strain W19 to promote plant growth and suppress the Fusarium wilt of banana was evaluated in both pot and field experiments. The results showed that application of BIO significantly promoted the growth and fruit yield of banana while suppressing the banana Fusariurn wilt disease. To further determine the beneficial mechanisms of the strain, the colonization of green fluorescent protein-tagged strain W19 on banana roots was observed using confocal laser scanning microscopy and scanning electron microscopy. The effect of banana root exudates on the formation of biofilm of strain W19 indicated that the banana root exudates may enhance colonization. In addition, the strain W19 was able to produce indole-3-acetic acid （IAA）, a plant growth-promoting hormone. The results of these experiments revealed that the application of strain W19-enriched BIO improved the banana root colonization of strain W19 and growth of banana and suppressed the Fusarium wilt. The PGPR strain W19 can be a useful biocontrol agent for the production of banana under field conditions.