Effect of activation methods for bovine oocytes after intracytoplasmic injection

A principal nuclear transfer procedure is to inject a donor cell into the perivitelline space in an enucleated oocyte and then electric fusion is performed (cell fusion method). The effects of activation methods in reconstructed oocytes for the serum-starved somatic cell cloning procedure were investigated in this study by means of intracytoplasmic injection (i.c.i.). Bovine oocytes were enucleated at 18-22 h for in vitro maturation, and subsequently the nucleus of cumulus cell collected from Japanese Black Bulls (JBCC) after 5-7 days of starved culture was injected into the recipient cytoplast with a piezo-micromanipulator. At 1 h after i.c.i., reconstructed oocytes were stimulated with ethanol (ET) or calcium ionophore (CaI) as the first activation treatment, followed by cycloheximide (CHX) or 6-dimethylaminopurin (DMAP) treatment as the second activation. In the experiment on the first activation method, the proportion of reconstructed oocytes developing to the blastocyst stage was significantly (p<0.01) higher in the ET activation method than that with CaI (10.5% and 4.7%, respectively). And the experiment on the second activation method after ET treatment showed similar proportions of blastocyst development in both CHX and DMAP treatments (5.9% and 2.8%, respectively). The present results indicated that combined activation treatment with ET and CHX was efficient for reconstructed bovine oocytes by i.c.i.     

In vitro but not in planta encapsidation of Rice gall dwarf virus core particles by the outer capsid P8 protein of Rice dwarf virus expressed in transgenic rice plants

Transencapsidation of the Rice gall dwarf virus (RGDV) inner core by the Rice dwarf virus (RDV) outer capsid P8 protein was examined in vitro and in planta. When RGDV core particles were incubated with an extract from RDV P8-transgenic rice leaf tissue, RDV P8 encapsidated the RGDV core particles to form double-shelled virus-like particles in vitro. In contrast, when RDV P8-transgenic rice plants were inoculated with RGDV, progeny RGDV particles contained RGDV P8 but RDV P8 was not detectable in the virions. No significant differences were found in acquisition by the vector insects and subsequent transmission rates between RGDV infecting nontransgenic rice plants and those infecting RDV P8-transgenic rice plants. These results indicate that mechanisms of and/or requirements for interactions between P8 and the inner core particles of phytoreoviruses differ between in vitro and in planta.     

Temporal and spatial dynamics of an old-growth beech forest in western Japan

Dendrochronological approaches enable us to understand forest stand dynamics by estimation of disturbance history and age structure. The present study was conducted in an old-growth beech forest in a forest reserve in western Japan. Increment cores were taken for tree ring analysis from all canopy trees in a 50 m × 130 m study plot. Radial growth release criteria were developed to identify significant growth releases in each tree ring series and to characterize the disturbance history of the study site. The age structure of the forest was indicative of continuous establishment by Fagus crenata and simultaneous establishment by Magnolia obovata. A variety of low-intensity disturbances were identified in each decade, especially after the 1900s, but the occurrence of high-intensity catastrophic disturbance was rare, and likely played an important role in maintaining species diversity in the existing forest canopy. The results also suggest that F. crenata regenerates gradually before and after both large- and small-scale disturbances, whereas M. obovata and Betula grossa regenerate only after large-scale catastrophic disturbances.     

Increased ectopic fat cells in the longitudinal muscularis layer of the oviduct isthmus in obese Japanese Black cows

In obese humans, mesenchymal stem cells differentiate to become ectopic fat cells in muscles. These ectopic fat cells inhibit the contraction of vascular smooth muscles. Stem cells have been recently identified in the human oviduct, a structure important in reproduction. We therefore investigated the number of Oil Red O (ORO)‐positive cells in the oviducts of control Japanese Black cows (n = 6; body condition score [BCS], 3.0 on a 5‐point scale) compared to those with diet‐induced obesity (n = 5; BCS, 4.0). We stained the ampulla and isthmus collected on the second day after ovulation with ORO and then counted the positive cells in each layer in 10 cross‐sections of the ampulla or isthmus. The obese group (23.4 ± 3.4 in the 10 sections) had larger numbers of ORO‐positive cells in the longitudinal muscularis of the isthmus (P < 0.05) than did the control group (15.0 ± 2.4). ORO‐positive cells were also observed in all other layers of the isthmus and ampulla; however, the number of cells in these layers did not differ significantly between obese cows and controls. Whether this observed increase in ORO‐positive cells in the oviducts of obese cows affects their reproduction warrants further study.     

How do visual and auditory cues influence calf stress reactions at weaning?

We investigated the effects of visual and auditory information related to cow and calf stress responses at weaning. Three weaning conditions were established with physical separation only (PS), physical and visual separation (VS), and physical, visual and auditory separation (VAS). Stress responses such as vocalization and self‐grooming, and changes of three maintenance behaviors (lying, walking and grazing) were recorded for experimental cows and calves in the respective conditions. Comparison of the data revealed that the cow responses differed slightly among groups. However, the peak frequency of vocalization was higher in the PS calves than in those of either VS or VAS. Self‐grooming frequency was significantly different in calves and was high in the order of VAS, VS and PS calves. Furthermore, the percentage of time spent lying down was greater, and that of walking was less, in VAS calves than in either VS or PS calves. These results demonstrate that different information at weaning might strongly affect the stress responses of calves but not those of cows. Relative impacts of different types sensory information on stressors must be considered to plan animal‐friendly weaning.     

Coexistence of anadromous and lacustrine life histories of the shirauo, Salangichthys microdon

The environmental history of the shirauo, Salangichthys microdon, was examined in terms of strontium (Sr) and calcium (Ca) uptake in the otolith, by means of wavelength dispersive X‐ray spectrometry on an electron microprobe. Anadromous and lacustrine type of the shirauo were found to occur sympatric. Otolith Sr concentration or Sr : Ca ratios of anadromous shirauo fluctuated strongly along the life‐history transect in accordance with the migration (habitat) pattern from sea to freshwater. In contrast, the Sr concentration or the Sr : Ca ratios of lacustrine shirauo remained at consistently low levels throughout the otolith. The higher ratios in anadromous shirauo, in the otolith region from the core to 90–230 μm, corresponded to the initial sea‐going period, probably reflecting the ambient salinity or the seawater–freshwater gradient in Sr concentration. The findings clearly indicated that otolith Sr : Ca ratios reflected individual life histories, enabling these anadromous shirauo to be distinguished from lacustrine shirauo.     

Overcoming the self-incompatibility of Raphanus sativus by application of plant hormones,amino acids and vitamines,and by temperature treatment of pollen

Summary Overcoming self-incompatibility by application of three kinds of plant hormones, sucrose, 3 kinds of amino acids and 2 kinds of vitamines was tested in cvs. Honbashi-taibyo Minowase (H-Mino) and Minowase (Mino) of Raphanus sativus. Effects differed between the cultivars. In H-Mino, BA (100 mg/l) and glutamic acid, folic acid and nicotinic acid (500 mg/l) resulted in higher fruit set and higher number of seeds per pollinated flower. In Mino, BA and NAA (100 mg/l) and glutamic acid and glycine (500 mg/l) induced a high number of seeds per pollinated flower. These chemicals, however, induced parthenocarpic fruit set, especially GA3. From the observation of pollen on stigmas washed with glutamic acid, it appeared that the pollen-tube penetrated into a papilla cell after 1 hour and openings of papillae and detached pollen grains and tubes were found after 2 hours as the result of successful pollentube penetration of papillae. Pollen was heated at 50°C for 30, 45 or 60 minutes, at 60°C for 15, 30 or 45 minutes and at 70°C for 10, 20 or 30 minutes prior to self-pollination. In H-Mino, 60 and 70°C were effective, and expecially 60°C for 15 or 30 minutes resulted a higher percentage fruit set and more seeds per fruit. In Mino, although 50–70°C were effective, the mean number of seeds per pollinated flower was lower than in H-Mino.     

Radiocesium transfer from Andosols to brown rice in the northern and northwest areas of Tochigi Prefecture,in the first 3 years following the 2011 Fukushima Daiichi nuclear power plant accident

ABSTRACT

Following the Fukushima Daiichi Nuclear Power Plant accident of 2011, the potential for radiocesium transfer from contaminated soils, such as Andosols, to agricultural crops became a significant concern. Andosols account for up to 70％ of paddy soils in the northern and northwest areas of Tochigi Prefecture, where the radiocesium concentration is 1000 Bq kg^?1 or greater in the soil of some fields. The present study was carried out in order to determine the phytoavailability of radiocesium in Andosols by comparing it with that of gray lowland soils in the first 3 years following the accident. The transfer factor (TF) tended to be higher in Andosols than in gray lowland soils, leading to higher radiocesium concentrations in brown rice grown in Andosols. The exchangeable potassium (Ex-K₂O) in Andosols was highly and negatively correlated with TF, followed by clay. The Ex-K₂O value was positively correlated with the clay/total carbon (T-C) value, suggesting that a high T–C ratio could weaken K₂O adsorption on clay mineral sites; hence, the low clay/T-C values can partially explain the relatively large TF values of Andosols. Samples with Ex-K₂O contents less than 200 mg kg^?1 and with low clay/T-C values showed striking decreases in TF values from 2011 to 2012. However, the decrease from 2012 to 2013 was quite small; radiocesium in these samples was potentially available for rice uptake for a long time, likely due to the reversible adsorption and fixation characteristics of allophane. Most gray lowland soil samples showed very low TF values over the 3 years of the study, except for those with TF values greater than 0.1 due to low Ex-K₂O and clay contents; the geometric mean (GM) value of TF was below 0.01 in 2012. The extraction of exchangeable radiocesium (Ex-Cs) with a 1 mol L^?1 ammonium acetate solution may not be an appropriate method for explaining the variability in radiocesium TF in Andosols. This is because the Ex-Cs value was significantly correlated with Ex-K₂O in Andosols, but not in gray lowland soils, indicating that Ex-K₂O explained this variability in relation to Ex-Cs.