Dispersal and persistence of genetically modified oilseed rape around Japanese harbors

Background, aim, and scope

The possibility of gene transfer from genetically modified oilseed rape (OSR) to its cultivated or wild relatives is of concern since its commercial cultivation, because of its potential weediness and impact on the environment. Introgression of modified genes can affect conservation of agricultural crops, because there are many cultivars and wild Brassicaceae that may cross with genetically modified OSR (Brassica napus) in Japan. Japan imports more than 2 million tons of OSR a year from Canada and other countries. Since volunteers of GM OSR were found around harbors in 2004, a lot of feral GM OSR was discovered in Japan. To consider the way how to keep domestic Brassicaceae from GM contamination, we surveyed and analyzed the dispersal and persistence of GM OSR around Japanese harbors. We present the cause and abundance of GM OSR in Japan by this paper.

Materials and methods

Survey of the feral OSR was performed several times a year at different seasons when domestic OSR either grows or does not around port areas. Detection of herbicide tolerance in feral B. napus was done by test papers that cross react with the modified gene product. Two kinds of herbicide tolerance (glyphosate and glufosinate) were tested.

Results

The feral B. napus were discovered around all 13 harbors that import rapeseeds from foreign countries. Genetically modified, herbicide-tolerant OSR were frequently found in the surveyed populations. Two kinds of herbicide-tolerant OSR (glyphosate- and glufosinate-tolerant) were discovered in a natural condition 40 km from port to an oil factory where 60,000 tons of OSR seed are processed a year. The cause of voluntary growth of OSR is seed spillage during transportation by trucks from harbors to oil factories and other processing facilities. Some of the feral OSR growing along the roadsides of transport paths exhibited perennial growth spilling their seeds around the places. Alteration of the generation of feral GM OSR was discovered for the first time in Japan as a result of this study. We studied the yearly change of feral OSR abundances focusing on Yokkaichi port over the 4 years since 2004. The rate of GM OSR increases year to year, and reaches nearly 90% in 2008.

Discussion

The possibility and consequences of gene transfer from the genetically modified OSR to domestic species (B. rapa and B. juncea) were discussed in relation to impact on domestic agriculture and on environments. Evolutional meaning of the gene transfer was also discussed with respect to the gene construct of GM OSR. This study shows the importance of another pathway of modified gene transfer to non-GM relative species by seed transportation in addition to pollen transfer from commercial cultivation of genetically modified OSR.

Conclusions and recommendations

We identified unintended dispersal and persistence of GM OSR around Japanese harbors that import OSR from Canada and other countries. Both glyphosate- and glufosinate-tolerant feral B. napus were discovered. The cause of volunteer OSR is spillage of the seeds during transportation by truck to oil factory. The feral GM OSR sometimes showed perennial growth in Japanese phonological conditions which are not observed for domestic Brassicaceae. In addition, we confirmed an alteration of generations by feral GM OSR in Japan. The possibility of cross pollination and GM gene introgression to domestic varieties can occur in these environments. To improve the situation, each responsible organization, company, administration, or government should establish measures how to stop the dispersal and persistence of GM OSR in nature. Also, the GM plant developers are responsible for revising this situation.     

Bioremediation of coastal areas 5 years after the Nakhodka oil spill in the Sea of Japan: isolation and characterization of hydrocarbon-degrading bacteria

Five years after the 1997 Nakhodka oil spill in the Sea of Japan, seven bacterial strains capable of utilizing the heavy oil spilled from the Nakhodka Russian oil tanker were isolated from three coastal areas (namely Katano Seashore of Fukui Prefecture, Osawa and Atake seashores of Ishikawa Prefecture) and the Nakhodka Russian oil tanker after a 5-year bioremediation process. All bacterial strains isolated could utilize long-chain-length alkanes efficiently, but not aromatic, and all of them were able to grow well on heavy oil. Using 16S rDNA sequencing, most of the strains were affiliated to Pseudomonas aeruginosa. Comparing between the year 1997 (at the beginning of bioremediation process) and the year 2001 (after 5 years of bioremediation), there was no significant change in morphology and size of hydrocarbon-degrading bacteria during the 5-year bioremediation. Scanning and transmission electron microscopic observations revealed that a large number of hydrocarbon-degrading bacteria still existed in the sites consisting of a variety of morphological forms of bacteria, such as coccus (Streptococcus and Staphylococcus) and bacillus (Streptobacillus). On the application of bioremediation processes on the laboratory-scale, laboratory microcosm experiments (containing seawater, beach sand, and heavy oil) under aerobic condition by two different treatments (i.e., placed the inside building and the outside building) were established for bioremediation of heavy oil to investigate the significance of the role of hydrocarbon-degrading bacteria on them. There was no significant bacterial activity differentiation in the two treatments, and removal of heavy oil by hydrocarbon-degrading bacteria in the outside building was slightly greater than that in the inside building. The values of pH, Eh, EC, and dissolved oxygen (DO) in two treatments indicated that the bioremediation process took place under aerobic conditions (DO: 1-6 mg/l; Eh: 12-300 mV) and neutral-alkaline conditions (pH 6.4-8) with NaCl concentrations of 3-15% (ECs of 45-200 mS/cm).     

Separation and recovery of carbon dioxide by a membrane flash process

A novel process for carbon dioxide (CO₂) separation, which was named a membrane flash process, was developed to realize an energy-saving technology and to substitute it for a conventional regenerator. The electric energy for CO₂ recovery in a membrane flash process using aluminum oxide and diethanolamine was lower than the thermal energy of the conventional chemical absorption process. Flashing at elevated temperature by the low temperature energy significantly reduced the electric energy and required much less membrane area. This process has potentiality of low cost capture of CO₂ when the low temperature energy, which is not available for other purposes, can be utilized to elevate flashing temperature.     

Concentrations of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and dioxin-like polychlorinated biphenyls in blood and breast milk collected from 60 mothers in Sapporo City, Japan

We measured the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), non-ortho polychlorinated biphenyls (non-ortho PCBs), and mono-ortho polychlorinated biphenyls (mono-ortho PCBs) in paired samples of blood and breast milk collected from 60 mothers in Sapporo City, Hokkaido Prefecture, Japan. The present study is one of the few studies in which PCDDs, PCDFs, and dioxin-like PCBs have been measured in blood and breast milk collected from 60 same mothers. Of these 60 mothers, 30 were primipara (mean: 30.1 years, median: 28.0 years) and 30 were multipara (mean: 32.2 years, median: 32.5 years). The arithmetic mean TEQ concentrations of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in blood and breast milk of the primiparous mothers were 9.0, 3.3, 5.7, and 0.4 pg TEQ g(-1) lipid, respectively, and 5.2, 2.2, 4.5, and 0.4 pg TEQ g(-1) lipid, respectively, with the total TEQ concentrations of these dioxin-like compounds being 9.3-42.9 (mean: 18.4, median: 17.3) and 7.0-41.1 (mean: 12.3, median: 11.4) pg TEQ g(-1) lipid, respectively. In the case of multiparous mothers, the arithmetic mean TEQ concentrations of these dioxin-like compounds in blood and breast milk were 7.1, 2.7, 5.3, and 0.4 pg TEQ g(-1) lipid, respectively, and 3.9, 1.7, 3.8, and 0.4 pg TEQ g(-1) lipid, respectively, with the total TEQ concentrations being 3.4-28.1 (mean: 15.5, median: 13.9) and 2.7-20.0 (mean: 9.8, median: 9.2)pg TEQ g(-1) lipid, respectively. The total TEQ concentrations of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in blood and breast milk of primiparous mothers in Sapporo City appeared to be generally lower than those recently surveyed throughout the greater area of Japan. Significant correlations were observed between age and the total TEQ concentrations of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in the blood of primiparae and multiparae, and significant correlations were also observed between age and the total TEQ concentrations of these dioxin-like compounds in the breast milk of primiparae and multiparae. The total TEQ concentrations of PCDDs, PCDFs, and dioxin-like PCBs in the blood of primiparous mothers showed a close correlation to those in their breast milk and also showed good correlations between the total TEQ concentrations of these dioxin-like compounds in the blood of multiparous mothers and those in their breast milk. The result of a comparison of the arithmetic mean concentrations of each congener of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in blood and breast milk indicated that the transfer of octachlorodibenzo-p-dioxin (OCDD) from the blood to the breast milk was lower than those of other congeners. In contrast, among mono-ortho PCBs congeners, the arithmetic mean concentrations of 2,3',4,4',5-pentachlorobiphenyl (PentaCB) (#118) and 2,3,3',4,4'-PentaCB (#105) in the breast milk were slightly higher than those in the blood, which suggested that breast-fed infants may be at higher risk from mono-ortho PCBs.     

Concentrations of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and non-ortho and mono-ortho polychlorinated biphenyls in blood of Yusho patients

We measured the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), non-ortho polychlorinated biphenyls (non-ortho PCBs) and mono-ortho polychlorinated biphenyls (mono-ortho PCBs) in blood collected from 279 Yusho patients living in Japan, 92 Yusho-suspected persons living in Japan and 127 normal controls living in Fukuoka Prefecture, and compared among the groups in terms the concentrations of these compounds. The total toxic equivalents (TEQ) concentrations of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in the bloods of Yusho patients and Yusho-suspected persons were 161.4 and 51.2 pg TEQ/g lipid, respectively and were about 3.5 and 1.1 times higher than those of normal controls, respectively. Although the TEQ concentrations of PCDDs, non-ortho PCBs, and mono-ortho PCBs among Yusho patients, Yusho-suspected persons and normal controls were nearly the same, the PCDFs levels of Yusho patients were significantly higher than those of Yusho-suspected persons and about 10.3 times higher than those of normal controls. In Yusho patients, PCDFs contributed about 65% to the total TEQ concentration. Among the PCDFs congeners for Yusho patients, the concentration of 2,3,4,7,8-pentachlorodibenzofuran (PeCDF) was about 11.3 times higher than that of normal controls. These findings indicated that Yusho patients even now, more than 34 years after the outbreak of Yusho, have much higher blood levels of 2,3,4,7,8-PeCDF in general than do unaffected persons.     

Identification of particles containing chromium and lead in road dust and soakaway sediment by electron probe microanalyser

Individual particles containing Cr and/or Pb and other major components were identified in road dust from a heavily used road (hereinafter 'heavy traffic road dust'), road dust from a residential area and soakaway sediment by electron probe microanalyser to locate their sources and carrier particles. Individual particles containing high levels of Cr and/or Pb (>or=0.2%) were identified using wavelength dispersive spectrometry (WDS) map analysis. Chromium, Pb and other major elements were then determined by means of a combination of WDS and energy-dispersive spectrometry in all identified particles, 50 particles containing neither Cr nor Pb from each type of road dust and soakaway sediment, and yellow road line markings. WDS map analysis revealed that many particles containing both Cr and Pb were present among the identified particles in heavy traffic road dust, whereas they were minor components in road dust from the residential area and soakaway sediment. The plots of X-ray intensities of Cr vs. Pb were linear for the identified particles containing both Cr and Pb in heavy traffic road dust, and the line closely fitted the plots for the three yellow road line marking samples. Individual particles were then classified using cluster analysis of element components. The results revealed that the adsorption of source materials or released metals onto soil minerals occurred in road dust and soakaway sediment, that the yellow road line markings were sources of Cr and Pb in heavy traffic road dust, and that materials containing Fe as a major component, such as stainless steel, were additional sources of Cr in both road dust and soakaway sediment.     

Determination of nonylphenol ethoxylates and octylphenol ethoxylates in environmental samples using 13C-labeled surrogate compounds

Alkylphenol polyethoxylates (APEOs) have been widely used as nonionic surfactants in a variety of industrial and commercial products. Typical compounds are nonylphenol polyethoxylates (NPEOs) and octylphenol polyethoxylates (OPEOs), which serve as precursors to nonylphenol (NP) and octylphenol (OP), respectively. NP and 4-t-OP are known to have endocrine disrupting effects on fish (medaka, Oryzias latipes), so it is important to know the concentrations of APEOs in the environment. Because the analytical characteristics of these compounds depend on the length of the ethoxy chain, it is necessary to use appropriate compounds as internal standards or surrogates. We synthesized two 13C-labeled surrogate compounds and used these compounds as internal standards to determine NPEOs and OPEOs by high-performance liquid chromatography (LC)-mass spectrometry. Method detection limits were 0.015 microg/L for NP (2)EO to 0.037 microg/L for NP(12)EO, and 0.011 microg/L for OP(3,6)EO to 0.024 microg/L for OP (4)EO. NPEO concentrations in water from a sewage treatment plant were less than 0.05-0.52 microg/L for final effluent and 1.2-15 microg/L for influent. OPEO concentrations were less than 0.05-0.15 microg/L for the final effluent and less than 0.05-1.1 microg/L for influent.     

Material flows of end-of-life home appliances in Japan

In Japan, the Law for the Recycling of Specified Kinds of Home Appliances came into effect in April 2001. This law aims to introduce appropriate waste treatment and promote the efficient use of resources so that we can achieve a sound material-cycle society. The law introduces a new framework for recycling that places obligations principally on the manufacturers and retailers of home appliances. Material flow analysis of this new framework is essential for evaluation of the system and for finding any problems that may exist. We therefore analyzed the material flows in facilities that recycle electrical home appliances. The mass balances in the recycling facilities were appropriately estimated. Then the potential destinations were surveyed and analyzed, even though the legal framework does not include flows after the recycling facilities. The potential destination after the facilities varies and depends on both the economy and the related regulations. Currently, considerable amounts of recovered materials are exported and recycled. However, the sustainability of these trades is unknown. Therefore, we should keep a close eye on the situation and make sure that our recycling system is adaptable to meet any future changes.     

A new protease assay system using gelatin thin film for monitoring indoor air quality

Airborne particulates have a major influence on the quality of indoor environments, which in turn affects human health. Both allergens and proteases are major etiological factors in allergic disease. Thus, the monitoring of environmental protease particulates is very important for the management of allergic disease. Protease-monitoring systems have been introduced in industry, but have not been applied to domestic settings, probably because of the high cost and many complex steps involved in these systems. In this study, we developed an indoor protease-monitoring system which can easily detect protease particulates without need for pre-treatment of dust samples or complex measuring instruments such as fluorescent plate reader. As a protease substrate, gelatin thin film (GTF) was adopted to evaluate small amounts of house dust particulates. The principle of this assay is based on the hydrolysis of GTF. Protease molecules spread from a particulate to GTF can hydrolyze the gelatin, thereby producing a spot on the GTF. This hydrolyzed spot visualized by staining was much larger than the particulate size, making visible to naked eye. The GTF method facilitated the observation of a subtle increase in protease particulates fallen on the GTF after the turnover of a used bed-quilt. The newly developed GTF method seems to be a suitable tool for protease monitoring at home.