Biochemical and Genetic Characterization of an Extracellular Poly(3-Hydroxybutyrate) Depolymerase from Acidovorax Sp. Strain TP4

To determine the properties of enzymes from bacteria that degrade polypropiolactone (PPL), we isolated 13 PPL-degrading bacteria from pond water, river water, and soil. Nine of these strains were identified as Acidovorax sp., three as Variovorax paradoxus, and one as Sphingomonas paucimobilis. All the isolates also degraded poly(3-hydroxybutyrate) (PHB). A PPL-degrading enzyme was purified to electrophoretical homogeneity from one of these bacteria, designated Acidovorax sp. TP4. The purified enzyme also degraded PHB. The molecular weight of the enzyme was estimated as about 50,000. The enzyme activity was inhibited by diisopropylfluorophosphate, dithiothreitol, and Triton X-100. The structural gene of the depolymerase was cloned in Escherichia coli. The nucleotide sequence of the cloned DNA fragment contained an open reading frame (1476 bp) specifying a protein with a deduced molecular weight of 50,961 (491 amino acids). The deduced overall sequence was very similar to that of a PHB depolymerase of Comamonas acidovorans YM1609. From these results it was concluded that the isolated PPL-degrading enzyme belongs to the class of PHB depolymerases. A conserved amino acid sequence, Gly-X₁-Ser-X₂-Gly (lipase box), was found at the N-terminal side of the amino acid sequence. Site-directed mutagenesis of the TP4 enzyme confirmed that ²⁰Ser in the lipase box was essential for the enzyme activity. This is the first report of the isolation a PHB depolymerase from Acidovorax.     

Determination of dissolved oxygen limitation in aerobic biofilm reactors

The concentration of dissolved oxygen (DO) strongly influences the performance of aerobic biofilm reactors because organic oxidation is limited by the availability of oxygen. However, it is not necessary to maintain a high DO level in the reactors in order to overcome this limitation. Excessive aeration wastes energy. Therefore, the determination of the onset of DO limitation against organic substrate removal in aerobic biofilm reactors is important for their effective operation. This study is aimed at developing an expression to determine the onset of DO limitation and hence to control the aeration system. The expression developed is as follows: , where S_b and C_b are the bulk concentrations of organic substrate and DO, respectively; D_ws and D_wc are the diffusion coefficients of organic substrate and oxygen in the reactors respectively; and R_b is an overall ratio of oxygen consumption to organic substrate removal in the reactors. The latter is the key parameter in the equation, and is determined by the characteristics of the substrate, biofilm, and reactor. In order to measure the value of R_b, the authors have developed a micro-biofilm reactor. The value of R_b was determined to be 0.13 (mg O₂ mg⁻¹ COD_cr) for glucose removal with this reactor. The equation has, subsequently, been verified with data from batch and continuous experiments.     

Effect of Poly(dioxolane) as Compatibilizer in Poly(ɛ-caprolactone)/Tapioca Starch Blends

The influence of poly(dioxolane) (PDXL), a poly(ethylene oxide-alt-methylene oxide), as compatibilizer on poly(ɛ-caprolactone) (PCL)/tapioca starch (TS) blends was studied. In order to facilitate blending; PCL, PDXL and TS must be blended together directly; so that PDXL is partially adhered at the TS surface as shown by scanning electron microscopy. The molecular weight effect of PDXL on the PCL/TS blends showed that mechanical properties of PCL/TS/PDXL blends from low molecular weight (M _n=10,000) and high molecular weight (M _n=200,000) PDXL were rather dependent on TS content. The enzymatic degradability of PCL/TS/PDXL blends using α-amylase increased as the TS content increased but was independent on the dispersion of tapioca starch in the PCL matrix.     

Biparental defence in a spider mite (Acari: Tetranychidae) infesting Sasa bamboo

Summary Field observations on the nesting pattern of Schizotetranychus celarius (Banks) (Acari: Tetranychidae) revealed that many individuals live gregariously in a united nest. Behavioural experiments recorded on video tape recorder and 16 mm film showed that adult males and females can effectively defend their nest and offspring from a phytoseiid predator, larvae of Typhlodromus bambusae Ehara (Acari: Phytoseiidae), through counterattacks. The male spider mite often killed the predator's larva and succeeded in defending offspring and nest completely. Comparison of the behaviour patterns recorded between a nest owner parent S. celarius and two kinds of intruders (predators and conspecific males or females) revealed conspicuous differences. I suggest that there is a kind of biparental care in S. celarius, and that the life-pattern of this species therefore fits the definition of subsociality originally framed for insects.     

Feasibility Study of Airbag Concept Applicable to Motorcycles Without Sufficient Reaction Structure

Objectives: An airbag system for motorcycle applications was developed and marketed in 2006 followed by many research projects on the system. In the airbag system, the bag should be supported during the kinetic energy–absorbing period of a rider in a collision. The previously developed system employed a configuration in which motorcycle structures support the airbag, such as a gauge unit and/or a steering structure. The supporting structure functions to receive the reaction force to hold the airbag during a crash to properly absorb the rider's kinetic energy. However, the previous system requires a larger area for this reaction structure and is applicable only to the motorcycles that can provide that area. To overcome this limitation, we propose an airbag system employing another concept. In this concept, the airbag does not use its vehicle structures as a reaction structure but uses the structures of an opposing vehicle, such as doors and/or pillars of an opposing vehicle. In this project, we aim to verify the effectiveness of the proposed system when installed in a motorcycle that cannot provide a larger area for the reaction structure.

Methods: In the system with this concept, it is assumed that the occupant protection performance is largely affected depending on impact configurations. Accordingly, full-scale motorcycle-to-car crash tests using 125 cm³ scooter-type models with and without the proposed system were conducted in various impact configurations. The 7 impact configurations specified in ISO 13232 were selected as the test configurations. Injury variables and injury indices of head, neck, chest, and abdomen were evaluated with the motorcyclist dummy.

Results: Injury variables and indices obtained from the crash tests with the airbag were compared to those of the baseline tests. In 2 impact configurations, the airbags were supported by the side structures of the opposing vehicle and performed to reduce the injury variable of head and/or chest compared to that of the baseline test.

Conclusion: Through the crash tests, beneficial protection effects of the airbag system were confirmed in particular impact configurations. No significant risk for the occupant due to the airbag was observed in the conducted crash tests. It was concluded that the proposed airbag system has feasibility to reduce rider injury in a collision of a motorcycle without sufficient reaction structure.     

Utilization of Broken Rice for the Production of Poly(3-hydroxybutyrate)

The feasibility of utilizing non edible rice (broken rice) for production of fine materials such as poly(3-hydroxybutyrate) (PHB) was considered as one of the alternative ways of keeping the environment clean for sustainable development. Thus, production of PHB from broken rice by simultaneous saccharification and fermentation (SSF) was investigated. During the SSF process, the rice (15% w/v) material was hydrolyzed to glucose, which was utilized by Cupriavidus necator for growth and production of PHB. The PHB content reached 38% at 58 h fermentation. The PHB had weight average molar mass (Mw) and polydipersity index of 3.82 × 10⁵ (g/mol) and 4.15, respectively. Differential calorimetric scan of the PHB showed a melting temperature (Tm) of 176 °C. Given that the PHB was a homopolymer (which consisted of (R)-3-hydroxybutyric acid monomers), it was thought that broken rice could be a raw material for production of both PHB and (R)-3-hydroxybutyric acid. This SSF process would not only help in the utilization of broken rice or non edible rice, but would also serve as a model for utilization of other raw materials that contain starch for production of PHB.     

Detection of Escherichia coli in a cattle manure composting process by selective cultivation and colony polymerase chain reaction

Livestock manure is suitable for use as a composting material. However, various intestinal microbes, such as Escherichia coli, are significant components of such manures. Thus, it is desirable that the level of intestinal microbes, and particularly opportunistic pathogens, in compost is inspected and counted regularly. The sensitivity and specificity of detection of E. coli in compost have been improved by selective cultivation followed by colony polymerase chain reaction (PCR) using the ECO primer. Indeed, the sensitivity of this method is higher than that of DNA extraction from compost and PCR. In this study, changes in numbers of E. coli present in a field-scale composting process over time was assessed using selective cultivation and colony PCR. Numbers of ECO-positive colonies after 24 h decreased, with a concomitant rise in compost temperature. ECO-positive colonies were not detected from 33 to 48 h. However, ECO-positive colony numbers increased beginning on day 4 and continuing until day 42. Thus, it seems likely that the high temperatures reached during the composting process did not affect E. coli numbers in the final compost. Additionally, selective cultivation followed by colony PCR using specific primers is an appropriate method of determining levels of cultivable pathogens in composted materials.     

Utilizing the Cyberforest live sound system with social media to remotely conduct woodland bird censuses in Central Japan

We have developed a system that streams and archives live sound from remote areas across Japan via an unmanned automatic camera. The system was used to carry out pilot bird censuses in woodland; this allowed us to examine the use of live sound transmission and the role of social media as a mediator in remote scientific monitoring. The system has been streaming sounds 8 h per day for more than five years. We demonstrated that: (1) the transmission of live sound from a remote woodland could be used effectively to monitor birds in a remote location; (2) the simultaneous involvement of several participants via Internet Relay Chat to listen to live sound transmissions could enhance the accuracy of census data collection; and (3) interactions through Twitter allowed members of the public to engage or help with the remote monitoring of birds and experience inaccessible nature through the use of novel technologies.     

Photodecomposition of humic acid and natural organic matter in swamp water using a TiO2-coated ceramic foam filter: Potential for the formation of disinfection byproducts

This paper reports on the photodecomposition of aqueous humic acid (HA) by a TiO₂-coated ceramic foam filter (TCF) reactor and on the potential for the formation of disinfection byproducts (DBPs) upon chlorination of the photocatalytically treated solutions. This photocatalytic reactor can also be applied to the removal of natural organic matter (NOM) in swamp waters. The proposed photocatalytic reaction system was operated as per standardized methodologies. First, the ability of the TCF to decompose HA (a representative compound of NOM) was evaluated from the changes in the total organic carbon (TOC) and UV₂₅₄ with the reaction time. Remarkably, TOC removal and UV₂₅₄ values ranging from 44% to 61% and from 60% to 83%, respectively, were achieved. The potential for the formation of DBPs (total trihalomethane and total haloacetic acid) by chlorination of the phototreated solution was strongly dependent on the TOC removal and UV₂₅₄ values in the solution. The degree of photodecomposition of NOMs in the swamp water samples and the DBP formation potential showed similar trends as in the case of the standard solutions containing HA. The method used in this study could be effectively used to evaluate the efficiency of TCF for reducing HA and NOM, while suppressing the formation of DBP products.