Development of Automatic Continuous Measurement System of Chemical Constituents in the Precipitation

Nowadays, acid rain is generally noticed as a global environmental problem. While acid rain has very much to do with the air pollutants, the relation between air pollution and chemical constituents in precipitation is not understood clearly yet. It is important to measure a variation of ion concentration in precipitation in short term for understanding the formation mechanism of acid rain. Therefore, an automatic continuous measurement system of chemical constituents in precipitation was developed and put into practical use in this study. The developed system was able to collect automatically every 1mm of precipitation and analyze major ions within 20 minutes.     

The first detection of Dicrocoelium chinensis sporocysts from the land snail Aegista vulgivaga in Gifu Prefecture, Japan

Trematodes of the genus Dicrocoelium are one of the most common parasites in ruminant animals; however, their life cycles in Japan are unclear. To find the sporocysts of D. chinensis in the natural field, we sampled 269 land snails (14 species) at a location with high level infection of sika deer in Gifu Prefecture, Honshu Island, Japan in autumn between 2017 and 2019. During the sampling period, we found mother sporocysts in the hepatopancreas of Aegista vulgivaga and Cyclophorus herklotsi. DNA barcoding based on the sequences of cytochrome c oxidase subunit 1 showed that the sporocysts from A. vulgivaga belonged to D. chinensis, indicating that this snail has potential as the first intermediate host of D. chinensis at this location.     

Mass balance of saline lakes considering inflow loads of rivers and groundwater: the case of Lake Issyk- Kul,Central Asia

This study aimed to elucidate the influence of inflow water on the salinity concentration process of a saline lake and the mass balance of Lake Issyk-Kul, a tectonic saltwater lake in Kyrgyzstan. Based on the survey results and meteorological data from 2012 to 2015, we analyzed the dissolved chemical composition loads due to water inflow. Then, we discussed the relationship between the increase in salinity and water inflow into the lake. Through the water quality analysis data, we used the tank model to estimate the river inflow and analyze the loads by the L-Q curve. The groundwater loads were then estimated from the average annual increase in salinity of the lake over a period of 30 a. The results suggest that Lake Issyk-Kul was temporarily freshened between about AD 1500 and 1800 when an outflowing river existed, and thereafter, it became a closed lake in AD 1800 and continued to remain a saline lake until present. The chemical components that cause salinization are supplied from the rivers and groundwater in the catchment area, and when they flow into the lake, Ca²⁺, HCO₃^- and Mg²⁺ precipitate as CaCO₃ and MgCO₃. These compounds were confirmed to have been left on the lakeshore as evaporite. The model analysis showed that 1.67 mg/L of Ca²⁺ and Mg²⁺ supplied from rivers and groundwater are precipitated as evaporite and in other forms per year. On the other hand, salinity continues to remain in the lake water at a rate of 27.5 mg/L per year. These are the main causes of increased salinity in Lake Issyk-Kul. Since Na⁺ and Cl^- are considered to be derived from geothermal water, they will continue to flow in regardless of the effects of human activities. Therefore, as long as these components are accumulated in Lake Issyk-Kul as a closed lake, the salinity will continue to increase in the future.     

Comparison of Salmonella enterica serovar Abortusequi isolates of equine origin by pulsed-field gel electrophoresis and fluorescent amplified-fragment length polymorphism fingerprinting

Equine paratyphoid is caused by Salmonella enterica serovar Abortusequi, and manifests mainly as abortion in the mare. We compared S. Abortusequi strains isolated in Japan and other countries using pulsed-field gel electrophoresis (PFGE) and fluorescent amplified-fragment length polymorphism (FAFLP) analysis. PFGE analysis of S. Abortusequi strains gave 21-27 fragments ranging in size from 33 to 602kb. Although two PFGE profiles were observed among the 20 S. Abortusequi isolates in Japan, the restriction fragments originating from the chromosome were common between the two profiles. The similarity index of the two profiles was 90.9%, while those between Japanese and five other S. Abortusequi strains were 29.8-37.5%. On the other hand, FAFLP analysis of S. Abortusequi strains generated 64-67 amplified fragments ranging in size from 100 to 400bp. One polymorphic fragment was observed among the 20 S. Abortusequi isolates in Japan. These data indicate the close relation of this agent in Japan. S. Abortusequi strains sharing a common ancestry might have been conserved in Japan.     

QTL analysis of black rot resistance in cabbage using newly developed EST-SNP markers

One hundred sixty-one EST-SNP markers were newly developed for analysis of QTLs for resistance to black rot caused by Xanthomonas campestris pv. campestris by determining EST sequences of a resistant line obtained from cabbage ‘Early Fuji’ and a susceptible broccoli line. A linkage map consisting of nine linkage groups was constructed with a total of 209 markers, including these new SNP markers and previously reported DNA markers. F₂ plants grown in a field for 1 month were inoculated by spraying bacteria of race 1, and disease severity of each plant was recorded. Three QTLs, i.e., QTL-1, QTL-2, and QTL-3, were detected on linkage group C2, C4 and C5, respectively. QTL-1, which showed the highest LOD score and additive effect, was again detected in another F₂ population used the next year, suggesting QTL-1 to be a major QTL. QTL-2 and QTL-3 could be minor QTLs influenced by environmental factors. The genomic region harboring QTL-1 showed synteny with a region from 5.3 to 7.4 Mb from the short arm end of chromosome 5 of Arabidopsis thaliana, which is rich in TIR-NBS-LRR family genes. The identified SNP markers in QTL-1 are considered to be useful in marker-assisted selection for black rot resistance in Brassica oleracea lines.     

Effect of co‐culture with intact embryos on development of bovine separated blastomeres

To improve embryo development in bovine separated blastomeres, we evaluated applicability of co‐culture with intact embryos. The morphological quality of blastocysts derived from separated blastomeres and rate of blastocyst formation were only slightly increased when the cells were co‐cultured with intact embryos, which did not provide significant differences when statistically analyzed. However, the cell count of inner cell mass (ICM), trophectoderm (TE) and total number of cells in Day 8 blastocysts were significantly higher when the cells were co‐cultured with the intact embryos than those with the cells cultured individually (P < 0.05). Transfer of four monozygotic pairs of blastocysts derived from the cells co‐cultured with intact embryos led to three pregnancies even when the blastomeres were produced by in vitro maturation and in vitro fertilization of oocytes collected by ovum pick‐up from elite cows. These results suggest that co‐culturing with intact embryos may enhance development of bovine separated blastomere.     

Primary analysis of DNA polymorphisms in the TRIM region (MHC subregion) of the Japanese quail,Coturnix japonica

Based on sequences of two cosmid clones from Japanese quail (Coturnix japonica, Coja), we confirmed that the syntenic cluster, GNB2L1～BTN1～BTN2, is located in the quail TRIM subregion of the quail major histocompatibility complex (MHC Coja) region. These cosmids also included four CjBG loci and one CjLEC locus; therefore, the quail TRIM subregion was thought to be adjacent to the BG/LEC subregion. We then identified three polymorphic markers – CjHEP21, CjTRIM39.2 and CjBTN2 – in the TRIM subregion that may be useful for the functional analysis of the MHC‐Coja region. We examined MHC‐Coja sequences from 321 individual quails sampled from 11 inbred strains, and we found eight alleles for each of the three genes – CjHEP21, CjTRIM39.2 and CjBTN2. These polymorphisms represent the first avian DNA markers in the TRIM subregion. Additionally, we discovered a quail‐specific VNTR (variable number of long tandem repeats, 133–137 bp) in intron 7 of CjBTN2. We identified 25 haplotypes in the sample of 321 quail; these haplotypes comprised combinations of all 24 alleles of the three polymorphic genes. We suggest that there are two recombination hotspots, one between each pair of adjacent loci. All strains, except AMRP, contained multiple haplotypes; the AMRP strain contained a single, apparently fixed haplotype.     

Downy mildew of busy lizzie caused by Plasmopara obducens in Japan

In June 2010 and March 2012, busy lizzie (Impatiens sultanii) plants in a glasshouse in Yamagata Prefecture and in nurseries in a plastic house in Shimane Prefecture, Japan were observed with yellowing of the adaxial surface of leaves or cotyledons and a white downy growth on the abaxial surface of leaves or cotyledons. We identified the causal pathogen as Plasmopara obducens based on the morphologies and rDNA-large subunit sequences. This new disease was named “downy mildew of busy lizzie” in Japan.     

Cesium-137 discharge into the freshwater fishery ground of grazing fish, ayu Plecoglossus altivelis after the March 2011 Fukushima nuclear accident

The Fukushima nuclear accident led to deposition of radionuclides into the freshwater fishery ground of ayu Plecoglossus altivelis. For the purpose of more comprehensive understanding of the radioactive contamination, cesium-137 (¹³⁷Cs) was analyzed in running water, muddy sediment, attached algae, and ayu using samples from three river systems with different levels of contamination within Fukushima Prefecture in summer and autumn 2011. The amount of ¹³⁷Cs contained in water as well as in sediment declined over an interval of two months, although the concentration in sediment samples was up to 10,000-fold higher than that in water samples. The changes in ¹³⁷Cs concentration of algae showed different courses among the three river systems, the concentration being greater in autumn in the river systems with high and moderate radioactive contamination. ¹³⁷Cs detected within a single stock of ayu appeared to be excreted rather rapidly with a calculated half-life of approximately 39 days, while the concentration ratio ranged from 10³ to 10⁴ l/kg, attaining higher levels than previously reported. These findings indicate that the most important pathway of ¹³⁷Cs into ayu is via intake of algae as food but that the retention time of ¹³⁷Cs inside ayu is relatively short.