Manipulated Mouse Embryos as Bioassay System for Water Quality Control

Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K⁺ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D‐O) water and Milli‐Q system (M‐Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA‐supplemented protein‐free culture media, whereas in EDTA‐ and protein‐free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M‐Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein‐free culture media were very low (0–7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M‐Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system.     

Detection of an isoform of alpha(1)-antitrypsin in serum samples from foals with gastric ulcers

The objective of this study was to find serum indicators of gastric ulcers in foals. By using two-dimensional electrophoresis of serum proteins, three distinct spots were detected in samples from foals with gastric ulcers detected endoscopically. One of them appeared with high frequency and was identified by partial digestion with trypsin and subsequent nano-electrospray ionisation-tandem mass spectrometry (nanoesi-ms/ms) analysis as an alpha(1)-antitrypsin. Western blot analysis, using an antibody against human alpha(1)-antitrypsin, revealed at least two bands, of molecular weight 58 kDa and 55 kDa, in the sera. The 55 kDa band was detected in 44 of 47 serum samples from foals with gastric ulcers, but in only three of 22 serum samples from healthy foals.     

Effects of land-use type and nitrogen addition on nitrous oxide and carbon dioxide production potentials in Japanese Andosols

Land-use type and nitrogen (N) addition strongly affect nitrous oxide (N₂O) and carbon dioxide (CO₂) production, but the impacts of their interaction and the controlling factors remain unclear. The aim of this study was to evaluate the effect of both factors simultaneously on N₂O and CO₂ production and associated soil chemical and biological properties. Surface soils (0–10 cm) from three adjacent lands (apple orchard, grassland and deciduous forest) in central Japan were selected and incubated aerobically for 12 weeks with addition of 0, 30 or 150 kg N ha^–1 yr^–1. Land-use type had a significant (p < 0.001) impact on the cumulative N₂O and CO₂ production. Soils from the apple orchard had higher N₂O and CO₂ production potentials than those from the grassland and forest soils. Soil net N mineralization rate had a positive correlation with both soil N₂O and CO₂ production rates. Furthermore, the N₂O production rate was positively correlated with the CO₂ production rate. In the soils with no N addition, the dominant soil properties influencing N₂O production were found to be the ammonium-N content and the ratio of soil microbial biomass carbon to nitrogen (MBC/MBN), while those for CO₂ production were the content of nitrate-N and soluble organic carbon. N₂O production increased with the increase in added N doses for the three land-use types and depended on the status of the initial soil available N. The effect of N addition on CO₂ production varied with land use type; with the increase of N addition doses, it decreased for the apple orchard and forest soils but increased for the grassland soils. This difference might be due to the differences in microbial flora as indicated by the MBC/MBN ratio. Soil N mineralization was the major process controlling N₂O and CO₂ production in the examined soils under aerobic incubation conditions.     

Classification of Azolla spp., section Azolla

Summary Azolla accessions (section Azolla) from the germplasm collections of the International Rice Research Institute and Washington State University were fingerprinted and classified by enzyme electrophoresis and leaf trichome morphology. A. filiculoides was enzymatically distinctive and also reliably identified by its prominent one-celled trichomes. Neotropical accessions labelled as A. filiculoides proved to be members of other species. Two groups of isolates were designated A. rubra, but those from Japan were identified as A. filiculoides. The A. rubra of Australia-New Zealand was biochemically unique and possessed less protuberant trichomes than A. filiculoides. A. microphylla, A. mexicana, and A. caroliniana were phenetically similar, but a. microphylla was identifiable from the others in the banding patterns of certain enzymes. A. mexicana and A. caroliniana were closely related enzymatically. The two-celled leaf trichomes of these three species were similar in size and shape.     

Evaluation of technologies for sequestering carbon in agricultural land by using organic filter underdrains

Technology for storing carbon in the subsoil of agricultural land using organic filter materials in underdrains was evaluated as an activity for sequestering CO₂ to mitigate global warming. First, the quantity of carbon remaining in wood chips and rice husks in underdrains was determined over 11 years after construction. Moreover, the quantity of CO₂ emissions from the construction of two underdrain types was calculated. Then, a survey was conducted to investigate the effect on greenhouse gas emissions of the filter material. The results indicate that greenhouse gas emissions would not increase. The quantity of storage carbon in the wood chip filter material of underdrains during their 15-year service life was estimated to be 6.76 tCO₂eq ha^?1. Rice husks, in contrast, were found ineffective in storing carbon. Therefore, the selection of the filter material for underdrains is important. The value of storage carbon in the wood chip filter material is similar to the amount of carbon sequestered by no-till farming, which is now being implemented as a global warming countermeasure based on soil management in agricultural land.     

Effects of oxygen tension in the gas atmosphere during in vitro maturation, in vitro fertilization and in vitro culture on the efficiency of in vitro production of mouse embryos

Effects of oxygen (O2) tension in the gas atmosphere during in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC) on the efficiency of in vitro production of mouse embryos were examined. Mouse oocytes recovered from large antral follicles were subjected to IVM in Waymouth medium for 15, 16 and 17 hr under 5 or 20% O2 and then subjected to IVF and IVC under 5 or 20% O2 tension. Lowering the O2 tension in the gas atmosphere for IVM from 20 to 5% improved the cleavage rate after IVF when the oocytes were subjected to IVM for 15 hr; however, no improvement in the cleavage rate was observed when the culture period for IVM was extended to 16 and 17 hr. Lowering the O2 tension to 5% for IVM and IVC improved the development of the cleaved oocytes to the blastocyst stage, regardless of the culture period for IVM. However, the O2 tension for IVF had no remarkable effect on the subsequent embryonic development. These results demonstrate that 5% O2 is superior to 20% O2 for IVM and IVC, and suggest that 20% O2 for IVM may delay oocyte maturation and/or the acquisition of fertilizability and impair the developmental competence of oocytes.     

Growth inhibition due to continuous cropping of dryland rice and other crops

Rice, corn, sorghum, mungbeans, and cowpeas were continuously grown on the same land at 2- to 6-week intervals between crops to determine effects on yield. Decline of growth and yield occurred during continuous cropping. Dryland rice, mungbeans, and cowpeas were affected most, corn slightly, and sorghum was not visibly affected.

Growth inhibition in dryland rice and mungbeans occurred after one or two crops; growing the same crop in rapid succession resulted il1low grain yields. The persistence of inhibitory effects was indicated. One crop rotation or 5 months of fallow removed only a part of the harmful effects.

The causal agent appears to have a specific affinity to the host crop. Root residues of the previous crop may serve as a source of the causal agent for the subsequent crop.

Biological agents were involved in growth inhibition of rnungbeans and dryland rice. In mungbeans, the inhibitory effects appeared to be directly dependent on microorganisms. It appeared that microorganisms were not the primary cause of harmful effects in dryland rice.     

Drainage and irrigation performance of hybrid ditches in converted paddy fields under winter wheat cultivation in Hokkaido

Paddy fields converted into winter wheat fields in Hokkaido, Japan, receive extremely high snowfall, creating a risk of flood damage to crops in spring due to waterlogging of snowmelt runoff and poor drainage. Meanwhile, in June there is relatively little rainfall, and a lack of moisture inhibits winter wheat growth. Therefore, we developed a method involving a series of 30-cm-deep ditches in agricultural fields to be used for drainage during the flood-prone period and for furrow irrigation during the dry period using water drawn from the canals that feed the paddy fields. The ditches are called ‘hybrid ditches’ as they are able to perform both drainage and irrigation functions. In this study, we investigated the optimal construction timing and spacing for hybrid ditches. We also evaluated their ability to improve the drainage and irrigation of winter wheat. We found that the optimal timing for digging hybrid ditches is immediately after sowing, and the inter-ditch spacing for irrigation should be 15 m or less. The hybrid ditches promoted increased soil temperature and healthy development of wheat plants by improving drainage during the flood-prone period. In addition, water was successfully supplied via the hybrid ditches to irrigate the fields in June. Under experimental conditions in which rainfall was excluded, grain yield was 10% higher and percent protein content was more than 1% point greater in the irrigated plot compared with the non-irrigated plot. Grain yield was also observed to increase by 3–29% in demonstration tests conducted at local farms. From these results, we conclude that hybrid ditches are capable of improving the growth and yield of winter wheat by improving drainage and providing irrigation in converted paddy fields in Hokkaido.