Abundance-dependent transmission of the pinewood nematode, <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis> (Nematoda: Aphelenchoididae), to the Japanese pine sawyer, <Emphasis Type="Italic">Monochamus alternatus</Emphasis> (Coleoptera: Cerambycidae), adult in its pupal chamber

The transmission ratio of the pinewood nematode (PWN), Bursaphelenchus xylophilus, to the emerging adult Japanese pine sawyer (JPS), Monochamus alternatus, in its pupal chamber is a determinant of the number of the nematodes carried by JPS beetles. To investigate the factors affecting the transmission ratio, we counted the number of the nematodes carried by 36 newly emerged JPS beetles and the number remaining in and around their pupal chambers, and then estimated the transmission ratio (the number of nematodes carried by a JPS adult as a percentage of the total number of nematodes aggregating in and around its pupal chamber). The total number of nematodes aggregating in and around a pupal chamber ranged from 0 to 19,041, and the number of nematodes carried by a beetle ranged from 0 to 18,920. The transmission ratio correlated with neither the water content of the wood around the pupal chamber nor the degree of wood discoloration caused by blue-stain fungus. The transmission ratio varied with the abundance of the nematodes aggregating in and around the pupal chamber. In pupal chambers with more than 1,000 nematodes, almost all the nematodes were transmitted to the beetle. However, in pupal chambers with fewer than 1,000 nematodes, the transmission ratio varied greatly, from 0 to 100%. These results suggest that aggregation of many PWNs in the pupal chamber might stimulate transmission of the PWNs to the JPS adult and that this abundance-related transmission might contribute to the large variation in the number of PWNs carried by the JPS beetle.     

Allele‐specific PCR typing and sequencing of the mitochondrial D‐loop region in four layer breeds

This study aimed to investigate the ability of single nucleotide polymorphism (SNP) haplotypes in chicken mtDNA for presumption of the origins of chicken meat. We typed five SNPs of the D‐loop region in mtDNA by allele‐specific PCR (AS‐PCR) in 556 hens, that is 233 White Leghorn (WL), 50 Dekalb‐TX35 (D‐TX), 140 Barred Plymouth Rock (BPR) and 133 Rhode Island Red (RIR) kept in the National Institute of Livestock and Grassland Science (NILGS, Tsukuba, Japan). Five haplotypes were observed among those chickens by AS‐PCR. WL, D‐TX, BPR and RIR displayed three, two, one and four SNP haplotypes, respectively. By a combination of the haplotypes by AS‐PCR and the breeds, these chickens were classified into 10 groups. After the D‐loop was sequenced in two chickens from every group (20 individuals), 15 SNP sites (including one insertion) and eight sequence haplotypes were observed. In conclusion, haplotype variation was observed in and among the layer breeds of the NILGS. This study demonstrates that SNP haplotypes in mtDNA should be appropriate for the presumption of the origins of chicken meat.     

One-step Generation of Phenotype-expressing Triple-knockout Mice with Heritable Mutated Alleles by the CRISPR/Cas9 System

Triple-knockout mice generated by the one-step CRISPR/Cas9 system were examined for the effects of multiple gene modifications on each phenotype and individual gene function. Sixty embryos were transferred, and 9 pups were obtained; all 9 pups had mutations on 3 loci, and 7 pups showed mutations in all-alleles. F0 mice showed knockout phenotypes or no protein expression of target genes simultaneously, and these mutations were normally inherited in the next generation.     

Different effects of ionic and non-ionic compounds on the freeze denaturation of myofibrils and myosin subfragment-1

The effects of non-ionic (sorbitol, maltose, trehalose) and ionic compounds (Na-glutamate, Na-acetate, Na-sulfate, ammonium sulfate) on freeze denaturation of myosin subfragment-1 (S-1) and of myofibrils were compared. Sugars, Na-glutamate and Na-acetate well suppressed the freeze denaturation of myofibrils as well as S-1 in a concentration dependent manner. Although sulfate suppressed freeze denaturation of S-1 irregularly, it accelerated myofibril denaturation. It was concluded that sulfate salts were useless as cryoprotectant for myofibrils. Stabilization extent by F-actin in frozen storage was much less than that in heating.     

Guidelines for soil conservation towards integrated basin management for sustainable development: A new approach based on the assessment of soil loss risk using remote sensing and GIS

Soil suspension in the Cidanau River of western Java, Indonesia, has increased recently, perhaps because of rapid environmental change in this watershed region. The objectives of this research are to assess soil loss risk using remote sensing and GIS and to develop effective guidelines for soil conservation in this watershed. To assess soil loss risk, a new soil loss model based on Universal Soil Loss Equation (USLE) was applied, in which C factor (crop management factor) was estimated using the perpendicular vegetation index (PVI); this was computed with satellite remote sensing data and used to simulate soil loss risk. The simulation showed that areas with highest risk of soil loss are on northern- and southern-facing hillsides with poor vegetation. Guidelines for soil conservation in the watershed were proposed: under these guidelines soil-loss risk is managed by evaluating the effectiveness of contour farming, belt farming, and mulch farming. Some recommended measures for soil conservation are as follows: (1) Green vegetation cover should be preserved as much as possible. (2) Vegetation coverage must be increased by forestation in steep sloped areas. (3) Belt farming and contour farming are recommended in areas with slopes under 100%, and mulch farming (more than 50% ground cover mulching is recommended) is desirable in areas with slopes over 100% and without green cover.     

Protective effect of sodium glutamate and sorbitol on thermal denaturation of chicken and croaker actomyosin

The thermal protective effect (E‐Value) of sodium glutamate (Na‐Glu) on chicken and croaker actomyosin (AM) was significant but decreased slightly with increasing concentration. The E‐value of chicken AM with Na‐Glu was 1.11 mol/L^?1 at 0–0.75 mol/L and decreased to 0.4 mol/L^?1 at 0.75–1.5 mol/L. Likewise, the E‐value of croaker AM with Na‐Glu was 1.51 mol/L^?1 at 0–1 mol/L and decreased to 0.2 mol/L^?1 at 1–1.5 mol/L. Sorbitol showed a steady thermal protective effect on chicken and croaker AM. The E‐values of chicken and croaker AM at 0 to 1.5 mol/L sorbitol were 0.71 mol/L^?1 and 0.59 mol/L^?1, respectively. The protective effect of sorbitol on both types of AM was greater than that of Na‐Glu. Higher concentrations of Na‐Glu with chicken and croaker AM gave a decrease in viscosity number and turbidity. Higher concentrations of Na‐Glu caused internal aggregation of AM and the protective effect of Na‐Glu was decreased. The results of this experiment confirmed that Na‐Glu and sorbitol have different modes of action and ability to control the thermal denaturation of chicken and croaker AM.     

Potential of preimplantation genomic selection using the blastomere separation technique in bovine in vitro fertilized embryos

Preimplantation genomic selection combined with an in vitro embryo production system is expected as a means of accelerating genetic improvement in cattle. While micromanipulation-based biopsy approaches are often used to collect embryonic cells for genetic testing, they require expensive equipment and sophisticated skills, hindering the adoption of this system. In the present study, to develop a simple method for preimplantation genomic selection using the blastomere separation (BS) technique in bovine in vitro fertilized embryos, we examined the accuracy of single nucleotide polymorphism (SNP) genotyping and optimal cryopreservation method in demi-blastocysts produced by the BS technique. We demonstrated reliable SNP genotyping using DNA derived from demi-blastocysts. We indicated a suitable equilibrium time in vitrification solution for demi-blastocysts and succeeded obtaining pregnancies by the transfer of vitrified demi-blastocysts. In conclusion, our findings suggest that the BS technique provides a simple method for preimplantation genomic selection in bovine in vitro fertilized embryos.     

Role of neck region in the thermal aggregation of myosin.

Carp dorsal myosin formed oligomers that retained ATPase activity upon heating. Cleavage of the oligomeric myosin at subfragment-1 (S-1)/rod junction released monomeric S-1 and rod, indicating that ATPase retaining myosin associated near the S-1/rod junction. The digest also contained rod oligomers. Heating a mixture of S-1 and rod generated neither ATPase retaining S-1 oligomers nor rod oligomers. Electron microscopic observation of the heated myosin revealed that some oligomers were formed by associating at the S-1/rod joining region, exhibiting a recognized double head, probably ATPase retaining oligomers. No myosin oligomers associated at the tail region were observed, thus, rod aggregation would be formed at its very restricted region near the S-1/rod junction. Based on the findings, we proposed that the neck structure is important in the thermal oligomerization process of myosin.     

不同种类与浓度的阴离子对菠菜镉吸收的影响

通过盆栽试验,研究了不同种类与浓度的阴离子（Cl-, SO42-）以及不同镉（Cd）水平对菠菜镉吸收的影响。结果表明,随着土壤中Cd浓度的增加,菠菜中的Cd浓度也随之增加。施Cd 1 mg/kg处理,菠菜中Cd浓度是对照的11～17倍;而施Cd 5 mg/kg处理,菠菜中Cd 浓度是对照的10～43倍。增施Cl- 可使土壤溶液中镉浓度增加,进而提高菠菜中镉浓度。施Cl- 5与10 mmol/kg,菠菜中Cd浓度分别为对照的1.45倍与2.31倍。施SO42-处理,则会降低菠菜中Cd浓度。施SO42- 5 mmol/kg ,Cd浓度仅为对照的0.77倍。土壤溶液中的Cd浓度与Cl- 浓度呈显著正相关。