Multilocus variable-number of tandem repeat analysis (MLVA) for Clostridium tyrobutyricum strains isolated from cheese production environment.

Clostridium tyrobutyricum is a gram-positive spore-forming anaerobe that is considered as the main causative agent for late blowing in cheese due to butyric acid fermentation. In this study, multilocus variable-number of tandem repeat (VNTR) analysis (MLVA) for C. tyrobutyricum was developed to identify the source of contamination by C. tyrobutyricum spores in the cheese production environment. For each contig constructed from the results of a whole genome draft sequence of C. tyrobutyricum JCM11008(T) based on next-generation sequencing, VNTR loci that were effective for typing were searched using the Tandem Repeat Finder program. Five VNTR loci were amplified by polymerase chain reaction (PCR) to determine their number of repeats by sequencing, and MLVA was conducted. 25 strains of C. tyrobutyricum isolated from the environment, raw milk, and silage were classified into 18 MLVA types (DI=0.963). Of the C. tyrobutyricum strains isolated from raw milk, natural cheese, and blown processed cheese, strains with identical MLVA type were detected, which suggested that these strains might have shifted from natural cheese to blown processed cheese. MLVA could be an effective tool for monitoring contamination of natural cheese with C. tyrobutyricum in the processed cheese production environment because of its high discriminability, thereby allowing the analyst to trace the source of contamination.

Detection and isolation of p-nitrophenol-lowering bacteria from intestine of marine fishes caught in Japanese waters.

To determine the existence of p-nitrophenol (PNP)-lowering bacteria in intestine of Japanese coastal fish, the gastro-intestinal contents were incubated in Brain Heart Infusion (BHI) broth and minimal medium (MM) broths containing 1 mmol/L PNP at 30 °C for 7 days. Among 26 samples of 19 fish species, 17 samples showed a decrease in PNP of 0.5-0.8 mmol/L in BHI broth, but no decrease was shown in MM broth. Eighteen PNP-lowering bacterial strains were isolated from four fishes. All of the strains were identified as Lactococcus lactis subsp. lactis. Three L. lactis strains JS1-3 isolated from Japanese seabass Lateolabrax japonicus showed the highest PNP-lowering activity (0.44 mmol/L). Optimum temperature and pH for the growth and PNP decreasing corresponded with the marine environment. These results suggested that marine fishes have PNP decreasing bacteria in their intestine. These bacteria might protect host fish from toxicities of PNP and PNP related compounds.

Reversible hydrogen storage property and structural analysis for face-centered cubic hydride Mg0.82Zr0.18H2 prepared by gigapascal hydrogen pressure method.

The face-centered cubic (fcc) type magnesium-zirconium hydride (Mg0.82Zr0.18Hx) was synthesized by means of the ultrahigh pressure (UHP) technique, which could generate 8 GPa of hydrogen pressure. The differential scanning calorimeter (DSC) measurements indicated that the fcc phase exhibited reversible hydrogen releasing and restoring properties under 0.5 MPa of hydrogen pressure. On the pressure-composition isotherms, the released and restored hydrogen capacities were estimated to be 3 approximately 3.5 wt %. The Rietveld analysis for synchrotron X-ray diffraction (XRD) data showed that the fcc phase had around 70 wt % mass fraction and was preserved without decomposition during hydrogen releasing and restoring cycles.