Water-flooded oil reservoirs have specific ecological environments due to continual water injection and oil production and water recycling. Using 16S rRNA gene clone library analysis, the microbial communities present in injected waters and produced waters from four typical water-flooded oil reservoirs with different in situ temperatures of 25°C, 40°C, 55°C and 70°C were examined. The results obtained showed that the higher the in situ temperatures of the oil reservoirs is, the less the effects of microorganisms in the injected waters on microbial community compositions in the produced waters is. In addition, microbes inhabiting in the produced waters of the four water-flooded oil reservoirs were varied but all dominated by Proteobacteria. Moreover, most of the detected microbes were not identified as indigenous. The objective of this study was to expand the pictures of the microbial ecosystem of water-flooded oil reservoirs.

Bacillus thuringiensis is the most widely used bacterial bio-insecticide, and most insecticidal crystal protein-coding genes are located on plasmids. Most strains of B. thuringiensis harbor numerous diverse plasmids, although the plasmid copy numbers (PCNs) of all native plasmids in this host and the corresponding total plasmid DNA amount remains unknown. In this study, we determined the PCNs of 11 plasmids (ranging from 2 kb to 416 kb) in a sequenced B. thuringiensis subsp. kurstaki strain YBT-1520 using real-time qPCR. PCNs were found to range from 1.38 to 172, and were negatively correlated to plasmid size. The amount of total plasmid DNA (∼8.7 Mbp) was 1.62-fold greater than the amount of chromosomal DNA (∼5.4 Mbp) at the mid-exponential growth stage (OD600 = 2.0) of the organism. Furthermore, we selected three plasmids with different sizes and replication mechanisms to determine the PCNs over the entire life cycle. We found that the PCNs dynamically shifted at different stages, reaching their maximum during the mid-exponential growth or stationary phases and remaining stable and close to their minimum after the prespore formation stage. The PCN of pBMB2062, which is the smallest plasmid (2062 bp) and has the highest PCN of those tested, varied in strain YBT-1520, HD-1, and HD-136 (172, 115, and 94, respectively). These findings provide insight into both the total plasmid DNA amount of B. thuringiensis and the strong ability of the species to harbor plasmids.

SummaryPlant‐parasitic nematodes are the most destructive group of plant pathogens worldwide and are extremely challenging to control. Some Bacillus thuringiensis crystal proteins are highly toxic to the plant‐parasitic nematode Meloidogyne incognita. In this study, the nematicidal crystal proteins Cry6Aa, Cry5Ba and Cry55Aa were tested against M. incognita to select the best toxin combination for its management. The results showed that a combination of Cry6Aa and Cry55Aa showed significant synergistic toxicity against M. incognita, and the highest synergistic effect (five times the expected toxicity of the two toxins calculated from their separate toxicities) was observed when they were combined in a 1:1 ratio. Furthermore, ligand blot analyses of the interaction between total proteins of M. incognita and the three toxins showed many different signal bands, indicating that there is a range of host proteins with which the toxins can interact. One explanation of the observed synergism is that the toxins damage the host in diverse ways, and they may thus act cooperatively and thereby show greater toxicity in combination. Our discovery provides an effective strategy for controlling M. incognita by using a combination of Cry6Aa and Cry55Aa.

Based on 454 pyrosequencing of 16S rRNA gene amplicons, microbial communities in samples collected from seven wells, six of which had positive paraffin deposition reduction and one with negative paraffin deposition reduction upon a microbial treatment designed for the prevention of paraffin precipitation, were analyzed. Microbial communities' structures were significantly different for the samples from the negative well and the positive wells. Microbes affiliated with Diaphorobacter belonging to β-Proteobacteria were predominant in the negative well, While γ-Proteobacteria-affiliated microbes of Pseudomonas and Enterobacter and Firmicute-affiliated Bacillus were shared and dominant in the positive wells. Microbes shared in the positive wells could be considered as potential candidates for investigations into microbial paraffin control. In addition, microbial activity of hydrocarbon-degradation and microbial products such as biosurfactants were proposed to be the main potential mechanisms for the microbial treatment for the prevention of paraffin precipitation.