Effect of sampling size on the determination of accurate pesticide residue levels in Japanese agricultural commodities

The uncertainty in pesticide residue levels (UPRL) associated with sampling size was estimated using individual acetamiprid and cypermethrin residue data from preharvested apple, broccoli, cabbage, grape, and sweet pepper samples. The relative standard deviation from the mean of each sampling size (n = 2(x), where x = 1-6) of randomly selected samples was defined as the UPRL for each sampling size. The estimated UPRLs, which were calculated on the basis of the regulatory sampling size recommended by the OECD Guidelines on Crop Field Trials (weights from 1 to 5 kg, and commodity unit numbers from 12 to 24), ranged from 2.1% for cypermethrin in sweet peppers to 14.6% for cypermethrin in cabbage samples. The percentages of commodity exceeding the maximum residue limits (MRLs) specified by the Japanese Food Sanitation Law may be predicted from the equation derived from this study, which was based on samples of various size ranges with mean residue levels below the MRL. The estimated UPRLs have confirmed that sufficient sampling weight and numbers are required for analysis and/or re-examination of subsamples to provide accurate values of pesticide residue levels for the enforcement of MRLs. The equation derived from the present study would aid the estimation of more accurate residue levels even from small sampling sizes.     

Gene response in rice plants treated with continuous fog influenced by pH,was similar to that treated with biotic stress

Background

Throughout Asia, including Japan, rice plants are cultivated in a wide range of areas from lowlands to highlands and are frequently exposed to fog, including acid fog. Some physiological studies have shown that acid fog can be a stress factor for plants. We analyzed the gene expression profiles of rice plants treated with artificially prepared simulated acid fog (SiAF) or simulated neutral fog (SiNF) for 1 or 7 days.

Results

Microarray analysis results suggested that both the SiAF and the SiNF treatments induced the expression of genes involved in the defense and stress responses in rice plants. Induction of such genes was detected in plants treated with SiAF for 1 day, and the number of induced genes increased in plants treated with SiAF for 7 days. The genes for defense and stress responses were also induced by SiNF for 7 days, although they were not induced by SiNF for 1 day. The gene expression profiles of the SiAF-treated and the SiNF-treated plants were compared to those of plants treated with other stress factors. The comparison revealed that both SiAF and SiNF treatments have similar effects to biotic stresses and ozone stress. The genes encoding NADPH oxidase and germin, which function in apoplasts, were also induced by SiAF, SiNF and biotic stresses.

Conclusions

These findings suggest that both the SiAF and the SiNF treatments may result in oxidative stress through the apoplastic production of reactive oxygen species.     

Optimization of in vitro embryo production and zygote vitrification for the indigenous Vietnamese Ban pig: The effects of different in vitro oocyte maturation systems

The Vietnamese Ban pig is a precious genetic resource that needs to be preserved. In vitro embryo production from in vitro matured (IVM) oocytes is an important tool for the utilization of cryopreserved porcine sperm. The aim of this study was to compare two media for the IVM of Ban pig oocytes. Immature oocytes were subjected to IVM either in a non‐defined (TCM‐199 + pig follicular fluid) or in a defined base medium (POM + epidermal growth factor). At the end of IVM, the oocytes were in vitro fertilized (IVF) with frozen Ban sperm. Ten hours after IVF, the oocytes were either subjected to orcein staining to check fertilization and maturation status or cultured in vitro for 7 days. There was no difference between the two IVM media in terms of percentages of oocyte maturation and blastocyst production. However, the percentage of male pronuclear formation after IVF and the total cell numbers in blastocysts were higher with the defined system. Zygotes obtained by the two IVM systems survived vitrification at similar rates. In conclusion, the two IVM systems were both effective for the production of Ban pig embryos; however, better embryo quality was achieved with the defined one.     

Abstracts of Nipon Dojo-Hiryogaku Zasshi

We investigated changes in radiocesium concentrations in a Japanese chestnut (Castanea crenata Sieold &; Zucc.) orchard in Ibaraki prefecture for 3 years after the Tokyo Electric Power Company’s Fukushima Daiichi nuclear power plant accident in March 2011. The radiocesium concentrations in the aboveground organs of Japanese chestnut trees were almost the same, while the concentration in the roots was the lowest among all the organs investigated. The concentration of radiocesium decreased exponentially for 3 years in nuts, leaves and current shoots. The radiocesium concentrations in soils were higher in the surface layer, and the trend of an annual decrease in radiocesium in the soils was similar to that of the natural decay of radiocesium. The transfer factor of radiocesium from soils to nuts of Japanese chestnut decreased annually. These results suggest that radiocesium adhered directly to the aboveground organs of Japanese chestnut trees in March 2011, and that the accumulation of radiocesium in nuts is mainly due to radiocesium transfer from the branches and trunk to nuts several years after the nuclear power plant accident.     

Protozoan predation of Escherichia coli in hydroponic media of leafy vegetables

Multiple outbreaks of food poisoning associated with fresh vegetable consumptions have occurred in many countries. Numerous reports have described human pathogenic bacteria, such as Escherichia coli O157:H7 and Salmonella spp., that can internalize into fresh vegetables via root or leaf surfaces. While attempting to obtain the threshold concentration of internalization of E. coli inoculated into hydroponic medium during vegetable cultivation, we observed a rapid decrease in E. coli numbers. In the present study, we determined that the rapid decline in E. coli was not due to a physiological change into a viable but non-culturable (VNC) state. The population crash was instead caused by true bacterial death, as the rapid descent was also confirmed by micro-colony fluorescence in situ hybridization, a culture-independent method that can detect VNC cells. We next monitored the number of E. coli inoculated into intact or filter-sterilized hydroponic medium after cultivation of various types of plants. We found that the number of E. coli in intact hydroponic medium decreased markedly, whereas the level in filter-sterilized hydroponic medium was completely unchanged. This result suggests that biotic factors were present that could be eliminated by filtering. Robust predation of E. coli by protozoa (ciliates and flagellates) was observed using fluorescently labeled bacteria incorporated into the hydroponic medium. Finally, morphological identification of flagellates by scanning electron microscopy revealed the presence of a species of Stramenopiles. These findings suggest the importance of protozoa as bacterial feeders in hydroponic systems and hence the use of these organisms as potential control agents of human pathogenic bacteria.     

Nitrogen metabolism of paddy rice at heading

The ears of paddy rice at boot stage, that is, just before heading, are being imbeded in the boot leaves' sheaths and are known to produce much guttation li:juid from their spikelets.     

Changes in fatty acid composition with age and environment in different types of peat profiles in Japan

Fatty acid composition of the crude lipid fraction of peat was investigated using several typical peat profiles in Japan. Fatty acid composition varied with the peat layers accumulated in a peat profile since 32,000 years BP. Deposition of long-range transported volcanic ash tephras also affected the composition remarkably due to the acceleration of decomposition. Fatty acid composition differed among high moor, transitional moor, and low moor peat profiles sampled in several locations in Hokkaido, Japan. The difference in the plants involved in the formation of peat was considered to be a very important factor determining the fatty acid composition. For example, arachidic acid was considered to be a good indicator for the presence of reed, while the percentage of stearic acid tended to be high in the high moor peat profiles dominated by sphagnum. As the fatty acids with a longer chain length were more stable than those with a shorter chain length, the percentage of longer fatty acids (C24 and C26) tended to increase and that of shorter fatty acids (CI4 and CI6) to decrease in the lower peat profiles. The drying of peatland also exerted a similar effect on the fatty acid composition.     

Incorporation of 14C into methionine as an intermediate for phytosiderophore biosynthesis in barley

In order to verify the precursory role of methionine (Met) in the biosynthesis of the mugineic acid family of phytosiderophores (MAs), feeding experiments of ¹⁴C-Iabeled compounds to barley (Hordeum vulgare L. cv. Minorimugi) roots grown hydroponically were conducted. When both ^l4C-Glucose (Glc) and unlabeled Met were fed to segmented roots, ¹⁴C was incorporated into Met and MAs in the roots. Molar-radioactivity of Met was higher than that of the amino-butanoic-acid unit in MAs in the roots. When ^l4C-Glc and unlabeled homoserine (Hse) were fed to decapitated roots, l4C was incorporated into Met but not into Hse. Therefore, it was considered that Hse might not be a major precursor of MAs. In addition, ¹⁴C was incorporated into Met and MAs in the roots when both ¹⁴C-glycerol (Gol) and unlabeled Met were fed to segmented roots. It is suggested that MAs may be synthesized from Glc via Met, bypassing Hse, and that the MAs biosynthesis may involve an unknown pathway associated with Gol and leading to Met.     

Effect of calcium concentration in nutrient solution on development of bacterial wilt and population of its pathogen Ralstonia solanacearum in grafted tomato seedlings

We examined the effect of the calcium (Ca) concentration in the nutrient solution on the development of bacterial wilt and the population of its pathogen Ralstonia solanacearum in tomato (Lycopersicon esculentum Mill. cv. Momotarou) seedlings grafted onto the rootstock of a highly resistant cultivar (cv. Hawaii 7998). The grafted seedlings were cultured in a nutrient solution containing Ca at concentrations of 0.4, 4.4, and 20.4 mm, and inoculated with the pathogen by stem puncture at the base of the stem of the rootstock, and the disease incidence was recorded for a period of 21 d. In another experiment, xylem exudates were collected from decapitated scions of the Ca-treated seedlings 5 d after inoculation, and the population of the pathogen in the exudates was counted by plating on a selective medium. The grafted tomato seedlings were highly resistant to bacterial wilt, when cultured in the nutrient solution containing Ca at 20.4 mm. The population of the pathogen in the xylem exudates decreased with increasing concentration of Ca in the solution. However, even in the presence of Ca at a high concentration, infection with the virulent pathogen was observed in the xylem of the scion.