Development of a Practical Method to Detect Noroviruses Contamination in Composite Meals

Various methods to detect foodborne viruses including norovirus (NoV) in contaminated food have been developed. However, a practical method suitable for routine examination that can be applied for the detection of NoVs in oily, fatty, or emulsive food has not been established. In this study, we developed a new extraction and concentration method for detecting NoVs in contaminated composite meals. We spiked NoV-GI.4 or -GII.4 stool suspension into potato salad and stir-fried noodles. The food samples were suspended in homogenizing buffer and centrifuged to obtain a food emulsion. Then, anti-NoV-GI.4 or anti-NoV-GII.4 rabbit serum raised against recombinant virus-like particles or commercially available human gamma globulin and Staphylococcus aureus fixed with formalin as a source of protein A were added to the food emulsion. NoV-IgG-protein A-containing bacterial complexes were collected by centrifugation, and viral RNA was extracted. The detection limits of NoV RNA were 10–35 copies/g food for spiked NoVs in potato salad and stir-fried noodles. Human gamma globulin could also concentrate other NoV genotypes as well as other foodborne viruses, including sapovirus, hepatitis A virus, and adenovirus. This newly developed method can be used as to identify NoV contamination in composite foods and is also possibly applicable to other foodborne viruses.     

Molecular size as a limiting characteristic for bioconcentration in fish

The relationships between the bioconcentration factor (BCF) of chemicals in fish and their size, as characterized by molecular weight (MW), effective cross sectional diameter (Deff), and maximum diameter (Dmax) have been investigated using an experimental data set of 737 new and 441 existing chemicals monitored by the Japanese Chemical Substances Control Law (CSCL). Substances with BCF > or = 5000 (very high bioconcentration potential) typically have MW < 550, Deff < 1.1 nm and Dmax < 2.0 nm, respectively and the substances with BCF > or = 1000 (high bioconcentration potential) have MW < 550, Deff < 1.4 nm and Dmax < 2.9 nm, respectively Therefore, the previously suggested threshold values for Deff (0.95 nm) and Dmax (1.5 nm) used for discriminating between bioconcentrative and non-bioconcentrative substances were found to be somewhat small. We found that many substances with BCF > or = 1000 and Dmax > or = 1.5 nm have Deff < 0.95 nm.     

Studies on automated survey system of environmental chemicals by gas chromatography-mass spectrometry-computer III. Construction of an automated survey system and its application

An automated survey system on the chemicals in environments by a gas chromatography-mass spectrometry-computer system has been constructed of Three Dimensional Mass Chromatography, Modified Probability Based Matching method, and Self Training Interpretive and Retrieval method. And it was applied to analyze environmental materials, with interesting results.     

Foliage profiles of individual trees determine competition,self-thinning,biomass and NPP of a Cryptomeria japonica forest stand: A simulation study based on a stand-scale process-based forest model

A simulation study was carried out to investigate simultaneously the effects of eco-physiological parameters on competitive asymmetry, self-thinning, stand biomass and NPP in a temperate forest using an atmosphere–vegetation dynamics interactive model (MINoSGI). In this study, we selected three eco-physiological relevant parameters as foliage profiles (i.e. vertical distribution of leaf area density) of individual trees (distribution pattern is described by the parameter η), biomass allocation pattern in individual tree growth (χ) and the maximum carboxylation velocity (V_max). The position of the maximal leaf area density shifts upward in the canopy with increasing η. For scenarios with η < 4 (foliage concentrated in the lowest canopy layer) or η > 12 (foliage concentrated in the uppermost canopy layer), a low degree of competitive asymmetry was produced. These scenarios resulted in the survival of subordinate trees due to a brighter lower canopy environment when η < 4 or the generation of spatially separated foliage profiles between dominant and subordinate trees when η > 12. In contrast, competition between trees was most asymmetric when 4 ≤ η ≤ 12 (vertically widespread foliage profile in the canopy), especially when η = 8. In such cases, vertically widespread foliage of dominant trees lowered the opportunity of light acquisition for subordinate trees and reduced their carbon gain. The resulting reduction in carbon gain of subordinate trees yielded a higher degree of competitive asymmetry and ultimately higher mortality of subordinate trees. It was also shown that 4 ≤ η ≤ 12 generated higher self-thinning speed, smaller accumulated NPP, litter-fall and potential stand biomass as compared with the scenarios with η < 4 or η > 12. In contrast, our simulation revealed small effects of χ or V_max on the above-mentioned variables as compared with those of η. In particular, it is notable that greater V_max would not produce greater potential stand biomass and accumulated NPP although it has been thought that physiological parameters relevant to photosynthesis such as V_max influence dynamic changes in forest stand biomass and NPP (e.g. the greater the V_max, the greater the NPP). Overall, it is suggested that foliage profiles rather than biomass allocation or maximum carboxylation velocity greatly govern forest dynamics, stand biomass, NPP and litter-fall.     

Some properties of trypsin-like proteases extracted from the seaweedCodium fragile and their purification

Two enzymes were purified from the green algaCodium fragile, collected in 1988 at Fukuoka, Japan, by batch-wise ion-exchange extraction, affinity chromatography and ion-exchange high-performance liquid chromatography (HPLC) using Boc-Ala-Ala-Pro-Arg-pNA as a substrate. The molecular weights of the enzymes were estimated as 38 000 and 39 000, using gel filtration HPLC. The enzymes had the same optimal pH range of 7 to 9 for both activities, and an exclusively hydrolyzed peptide bond on the carboxyl-terminal side of theL-arginine of peptidep-nitroanilides. The ratios of the enzymatic activity for X-Arg-pNA to X-Lys-pNA were larger than 100. The enzymes exhibited 30 times higher activity toward Boc-Ala-Ala-Pro-Arg-pNA when compared with trypsin. The activities were strongly inhibited by diisopropyl phosphofluoridate (DFP), and partially inhibited by phenylmethylsulfonyl fluoride (PMSF), benzamidine, leupeptin and antipain. The isolated enzymes were presumed to be trypsin-like serine protease from their primary substrate specificities and inactivation behavior.     

Studies on automated survey system of environmental chemicals by gas chromatography-mass spectrometry-computer I. Retrieval system of mass spectra

A retrieval system of mass spectra based upon the Probability Based Matching method was studied in order to apply it to survey the chemicals in environments by a gas chromatography-mass spectrometry-computer system. As retrieval indices, peak missing probability and relative confidence value were proposed.     

Trends in waste plastics and recycling

Zero emission is an ambitious project aimed at the conversion of the conventional top-down or one-directional flow type production systems of modern society to those of a recycling type. The basic idea underlying this project is to understand in detail the material flow mechanism in a particular segment of society and to evaluate the transformability of the hitherto one-directional types of process into recycling types. The authors, members of the Zero-Emission Research Group of Japan, are investigating the material flow of plastics in Japan. The present work describes the features of this project, as well as the results obtained so far. Received: May 28, 1998 / Accepted: October 16, 1998     

Spatial distribution and source identification of wet deposition at remote EANET sites in Japan

Wet deposition of major ions was discussed from the viewpoint of its potential sources for six remote EANET sites in Japan (Rishiri, Happo, Oki, Ogasawara, Yusuhara, and Hedo) having sufficiently high data completeness during 2000–2004. The annual deposition for each site ranged from 12.1 to 46.6 meq m⁻² yr⁻¹ for nss-SO₄²⁻, from 5.0 to 21.9 meq m⁻² yr⁻¹ for NO₃⁻. The ranges of annual deposition of the two ions for the sites were lower than those for urban and rural sites in Japanese Acid Deposition Survey by Ministry of the Environment, Japan, and higher than those for global remote marine sites. Factor analysis was performed on log-transformed daily wet deposition of major ions for each site. The obtained two factors were interpreted as (1) acid and soil source (or acid source for some sites), and (2) sea-salt source for all the sites. This indicates that wet deposition of ions over the remote areas in Japan has a similar structure in terms of types of sources. Factor scores of acid and soil source were relatively high during Kosa (Asian dust) events in spring in western Japan. Back-trajectories for high-deposition episodes of acid and soil source (or acid source) for the remote sites showed that episodic air masses frequently came from the northeastern area of Asian Continent in spring and winter, and from central China in summer and autumn. This indicates a large contribution of continental emissions to wet deposition of ions over the remote areas in Japan.     

Metabolism and elemental composition of four oncaeid copepods in the western subarctic Pacific

Respiration rates and elemental composition (carbon and nitrogen) were determined for four dominant oncaeid copepods (Triconia borealis, Triconia canadensis, Oncaea grossa and Oncaea parila) from 0–1,000 m depth in the western subarctic Pacific. Across the four species of which dry weight (DW) varied from 2.0 to 32 μg, respiration rates measured at in situ temperature (3°C) increased with DW, ranging from 0.84 to 7.4 nl O₂ individual⁻¹ h⁻¹. Carbon (C) and nitrogen (N) composition of the four oncaeid species ranged from 49–57% of DW and 7.0–10.3% of DW, respectively, and the resultant C:N ratios were 4.8–8.3. The high C contents and C:N ratios were reflected by large accumulation of lipids in their body. Specific respiration rates (SR, a fraction of body C respired per day) ranged between 0.5 and 1.3% day⁻¹. Respiration rates adjusted to a body size of 1 mg body N (i.e. adjusted metabolic rates, AMR) of the four oncaeid species [0.6–1.1 μl O₂ (mg body N)^−0.8 h⁻¹ at 3°C] were significantly lower than those (1.7–5.1) reported in the literature for oithonid and calanoid copepods at the same temperature. The present results indicate that lower metabolic expenditure due to less active swimming (pseudopelagic life mode) together with rich energy reserve in the body (as lipids) are the characters of oncaeid copepods inhabiting in the epi- and mesopelagic zones of this region.     

Formation of estrogenic products from environmental phthalate esters under light exposure

Phthalate esters (PEs) have been suspected to be environmental endocrine disruptors and the detailed mechanism remains unclear. The activities of these chemicals can be enhanced through chemical modification under the environmental conditions. We demonstrate that PEs acquire unequivocal estrogenic activity by light exposure. Through UV exposure of an aqueous PE solution, one active photoproduct, identified as 4-hydroxyPE (PE-4OH) based on its characteristic UV and mass spectra, was detected in an estrogen receptor alpha-dependent transactivation assay. PE-4OH was effectively generated by UV 290 nm. The PE-4OH production accompanied H2O2 generation in a UV dose-dependent manner. Both PE and UV irradiation were indispensable in the generation of H2O2. Addition of H2O2 to the PE solution increased PE-4OH production under UV irradiation. The PE-4OH production was also observed in the PE reaction with the Fenton reagent generating hydroxyl radical without UV irradiation. The proposed mechanism for PE-4OH production based on these results is such that by PE-mediated photosensitization H2O2 is generated from O2 and H+ and decomposed to hydroxyl radical, thus oxidizing the PE benzene ring. The PEs-4OH are remarkably active estrogenic products of PEs and would be involved in ER-mediated endocrine disruption.