Production of Poly(3-hydroxyalkanoate)s by Pseudomonas putida Cultivated in a Glycerol/Nonanoic Acid-Containing Medium

The biosynthesis of poly(3-hydroxyalkanoate) (PHA) by Pseudomonas putida (JCM6160) cultivated in a medium containing glycerol, nonanoic acid, or a glycerol/nonanoic acid mixture as the sole carbon sources was investigated. The PHA content was ~20 % when glycerol was the carbon source. This relatively low content can be attributed to the glycerol end-cap effect and the absence of enzymes that can directly synthesize PHA from acetyl CoA, which is the major metabolite of glycerol. Fatty acids, containing even numbered carbons, are synthesized from acetyl CoA, and they can be used as substrates for PHA synthesis. However, this process also results in decreasing PHA content as fatty acids are siphoned off into other pathways. However, addition of 5 mM nonanoic acid into a 20 mM glycerol-containing medium dramatically increased the PHA content in P. putida, which was 1.3 times larger than the sum of the values found when glycerol and nonanoic acid were each used as the sole carbon source. The PHA, synthesized in the glycerol/nonanoic acid medium, contains 3-hydroxy alkanoate units that have 5, 6, 7, 8, 9, or 10 carbons. The units that contain the even numbered carbons are derived from fatty acids that were produced from glycerol; whereas, the PHA units with the odd numbered carbons are derived from nonanoic acid. Pentanoate units were also found in the polyester derived from glycerol and nonanoic acid, and must have been synthesized indirectly via β-oxidation of nonanoic acid with the assistance of glycerol because pentanoate units were not found in PHA when P. putida was cultivated in the presence of only nonanoic acid.     

Intracellular localization of vanadium in the fan worm Pseudopotamilla occelata

A very high level of vanadium was found to be contained in the polychaete worm Pseudopotamilla occelata collected in 1992–1993 from the Sanriku coast on the main island of Japan. The vanadium concentration (mean±SD=5500±1800 g g^-1 dry wt) in the worm's branchial crown which is composed of many bipinnate radioles was approximately 100 times higher than that (mean±SD=60±25 g g^-1 dry wt) in the trunk body. Electron probe X-ray microanalysis revealed that a large amount of vanadium was present in the outer potion of the epidermis of the bipinnate radiole. Analytical electron microscopy for a cryo-section of the bipinnate radioles indicated that vanadium was localized in electron-dense deposits in the apical portion of epidermal cells. From an examination of the fine structures, the locality of the electron-dense deposits were found to correspond to that of the apical vacuoles in the epidermal cells. It was concluded that most of the vanadium in P. occelata was contained in the vacuoles of the epidermal cells of the bipinnate radioles.     

Migration of C in the paddy soil-to-rice plant system after C-acetic acid breakdown by microorganisms below the plow layer

Migration of ¹⁴C derived from ¹⁴C-acetic acid was examined by using soils sampled from paddies in four administrative areas in Japan (Aomori, Yamanashi, Ehime and Okinawa) and rice plant in a tracer experiment to understand the fate of ¹⁴C in the paddy soil-to-rice plant system. The loss of ¹⁴C radioactivity levels derived from ¹⁴C-acetic acid was caused by soil microorganism breakdown. A part of the ¹⁴C fixation to soil was caused by microbial assimilation into the fatty acid fraction. ¹⁴C moved upward via two different types of ¹⁴C dynamics in soil: quick movement upward; and constant but slow movement upward. ¹⁴C was highly assimilated into the plant panicle and that was caused by the root-uptake and the transfer of ¹⁴C. Migration of ¹⁴C derived from ¹⁴C-acetic acid relied heavily upon changes of chemical forms and characteristics of ¹⁴C-compound as caused by microorganisms in soil.     

Numerical model for a watering plan to wash out organic matter from the municipal solid waste incinerator bottom ash layer in closed system disposal facilities

Bottom ash from municipal solid waste incineration (MSWI) is a main type of waste that is landfilled in Japan. The long-term elution of organic matter from the MSWI bottom ash layers is a concern because maintenance and operational costs of leachate treatment facilities are high. In closed system disposal facilities (CSDFs), which have a roof to prevent rainfall from infiltrating into the waste layers, water must be supplied artificially and its quantity can be controlled. However, the quantity of water needed and how to apply it (the intensity, period and frequency) have not been clearly defined. In order to discuss an effective watering plan, this study proposes a new washout model to clarify a fundamental mechanism of total organic carbon (TOC) elution behavior from MSWI bottom ash layers. The washout model considers three phases: solid, immobile water and mobile water. The parameters, including two mass transfer coefficients of the solid-immobile water phases and immobile-mobile water phases, were determined by one-dimensional column experiments for about 2 years. The intensity, period and frequency of watering and other factors were discussed based on a numerical analysis using the above parameters. As a result, our washout model explained adequately the elution behavior of TOC from the MSWI bottom ash layer before carbonation occurred (pH approximately 8.3). The determined parameters and numerical analysis suggested that there is a possibility that the minimum amount of water needed for washing out TOC per unit weight of MSWI bottom ash layer could be determined, which depends on the two mass transfer coefficients and the depth of the MSWI bottom ash layer. Knowledge about the fundamental mechanism of the elution behavior of TOC from the MSWI bottom ash layer before carbonation occurs, clarified by this study, will help an effective watering plan in CSDFs.     

Centralization of dairy farming facilities for improved economics and environmental quality

In Japan, most farm animal excreta has been stored directly on farmland. Runoff from this storage has often caused water pollution. Biogasification is anticipated as an important technology to manage excreta properly, but complex problems hinder its introduction. Economic aspects of management have been especially difficult for dairy farmers. For this study, structural problems regarding introduction of biogasification into dairy farming were identified. Subsequently, a desirable system of dairy farming including biogasification was suggested, and an evaluation model of the financial balance was constructed. A case study using current financial balances of several systems of dairy farming was evaluated using the constructed model and actual data. The systems were based on several policy alternatives including the suggested system mentioned above. Results show that a farmer can obtain sufficient income from a system featuring centralization of dairy housing and biogasification facilities and coordinated management by over six farmers.     

Assessment of ozone effects on local rice cultivar by portable ozone fumigation system in Hanoi, Vietnam

This study examined the effects of troposphere ozone (O3) on rice by developing a portable ozone fumigation system and applied to the field in Hanoi, Vietnam. The system was nearly identical to the open top chamber but designed to easily and cheaply assess the O3 sensitivity of local agricultural crops in the actual field. The rice (Oryza sativa L.) was exposed to ozone during the flowering stage at peri-urban experimental site of Hanoi. The fumigation protocols covered five different levels, namely; non-filtered air (ambient) as the control treatment, 32, 62, 85 and 113 ppb in 7-h mean. It was found that observed impacts corresponded well to similar open-top chamber studies in other Asian countries. The methodology and the system left a large room for improvement and called for very careful interpretation and pre-conditions (e.g. low ambient O3 concentration). However, the portable O3 exposure in the field might open a door to the less developed countries to implement the phytotoxic risk assessment of the local agricultural species.     

Ozonation as an efficient pretreatment method to alleviate reverse osmosis membrane fouling caused by complexes of humic acid and calcium ion

Humic acids (HA) didn’t cause obvious reverse osmosis (RO) membrane fouling in 45 h. Osmotic pressure (NaCl) affected slightly the RO membrane fouling behavior of HA. Ca²⁺ promoted aggregation of HA molecules and thus aggravated RO membrane fouling. Ozonation eliminated the effect of Ca²⁺ on the RO membrane fouling behavior of HA. The change of the structure of HA was related to its membrane fouling behavior. Humic acid has been considered as one of the most significant sources in feed water causing organic fouling of reverse osmosis (RO) membranes, but the relationship between the fouling behavior of humic acid and the change of its molecular structure has not been well developed yet. In this study, the RO membrane fouling behavior of humic acid was studied systematically with ozonation as a pretreatment method to control RO membrane fouling. Furthermore, the effect of ozone on the structure of humic acid was also explored to reveal the mechanisms. Humic acid alone (10–90 mg/L, in deionized water) was found not to cause obvious RO membrane fouling in 45-h operation. However, the presence of Ca²⁺ aggravated significantly the RO membrane fouling caused by humic acid, with significant flux reduction and denser fouling layer on RO membrane, as it was observed by scanning electron microscope (SEM) and atomic force microscope (AFM). However, after the pretreatment by ozone, the influence of Ca²⁺ was almost eliminated. Further analysis revealed that the addition of Ca²⁺ increased the particle size of humic acid solution significantly, while ozonation reduced the SUVA₂₅₄, particle size and molecular weight of the complexes of humic acid and Ca²⁺ (HA-Ca²⁺ complexes). According to these results and literature, the bridge effect of Ca²⁺ aggregating humic acid molecules and the cleavage effect of ozone breaking HA-Ca²⁺ complexes were summarized. The change of the structure of humic acid under the effect of Ca²⁺ and ozone is closely related to the change of its membrane fouling behavior.