Mouse in vivo-derived late 2-cell embryos have higher developmental competence after high osmolality vitrification and −80°C preservation than IVF or ICSI embryos

Mammalian embryos are most commonly cryopreserved in liquid nitrogen; however, liquid nitrogen is not available in special environments, such as the International Space Station (ISS), and vitrified embryos must be stored at −80°C. Recently, the high osmolarity vitrification (HOV) method was developed to cryopreserve mouse 2-cell stage embryos at −80°C; however, the appropriate embryo is currently unknown. In this study, we compared the vitrification resistance of in vivo-derived, in vitro fertilization (IVF)-derived, and intracytoplasmic sperm injection (ICSI)-derived mouse 2-cell embryos against cryopreservation at −80°C. The ICSI embryos had lower survival rates after warming and significantly lower developmental rates than the in vivo and IVF embryos. Further, IVF embryos had a lower survival rate after warming, but a similar rate to the in vivo embryos to full-term development. This result was confirmed by simultaneous vitrification of in vivo and IVF embryos in the same cryotube using identifiable green fluorescent protein-expressing embryos. We also evaluated the collection timing of the in vivo embryos from the oviduct and found that late 2-cell embryos had higher survival and developmental rates to full-term than early 2-cell embryos. Some early 2-cell embryos remained in the S-phase, whereas most late 2-cell embryos were in the G2-phase, which may have affected the tolerance to embryo vitrification. In conclusion, when embryos must be cryopreserved under restricted conditions, such as the ISS, in vivo fertilized embryos collected at the late 2-cell stage without long culture should be employed.     

The effect of frequent milk feeding on abomasal curd formation of Holstein calves

In order to understand the effects of the automatic milk replacer feeding system on calf health, we examined the effect of frequent milk feeding on curd formation in the abomasum using ultrasonographic imaging. Eight male Holstein calves were divided into a milk-replacer group and a fresh milk group. Calves were fed twice a day to 12 days after birth (Period A). From 13 days, calves were fed six times a day (Period B). The abomasal fluid was taken by paracentesis. In both periods A and B, the fresh milk group formed bigger curds faster than the milk replacer group. The curd score of the milk replacer group in period B at 2 h was significant lower than those of both groups in period A. The pH in the abomasum was lower in period B than in period A. We could not identify the location of the abomasum in the milk replacer group during period B at almost time points. Our experiment suggests the possibility that frequent feeding of milk replacer causes incomplete hydrolysis of κ-casein as well as curd formation, thus reducing the digestibility compared to the feeding of the milk replacer twice per day or frequent feeding of fresh milk.     

Characterization of fucoxanthin and fucoxanthinol esters in the Chinese surf clam, Mactra chinensis

Fucoxanthin and fucoxanthinol fatty acid esters in the clam, Mactra chinensis, were characterized on the basis of 1H NMR and FAB-MS spectra. 1H NMR revealed that the hydroxy group at C-3 in fucoxanthin and fucoxanthinol was acylated. 3'-O-Acylated compounds such as fucoxanthinol 3'-ester or fucoxanthinol 3,3'-diester were not found in the clam. The fatty acids esterified with fucoxanthin and fucoxanthinol were identified as C24:6, C22:5, C22:6, C20:5, C20:0, C20:1, C18:0, C18:1, C16:0, C16:1, and C14:0 by FAB-MS data.     

Isolated chloroplast division machinery can actively constrict after stretching

Chloroplast division involves plastid-dividing, dynamin, and FtsZ (PDF) rings. We isolated intact supertwisted (or spiral) and circular PDF machineries from chloroplasts of the red alga Cyanidioschyzon merolae. After individual intact PDF machineries were stretched to four times their original lengths with optical tweezers, they spontaneously returned to their original sizes. Dynamin-released PDF machineries did not retain the spiral structure and could not be stretched. Thus, dynamin may generate the motive force for contraction by filament sliding in dividing chloroplasts, in addition to pinching-off the membranes.     

Effects of livestock grazing on the spatial heterogeneity of net soil nitrogen mineralization in three types of Mongolian grasslands

Purpose

Small-scale soil heterogeneity relates to productivity and biodiversity and is crucial to understand. Soil heterogeneity could be affected by vegetation structure, and large mammal grazers could modify it through herbivory and excretion. The objective is to clarify the effects of livestock grazing on the small-scale (～3 m) soil heterogeneity in three types of Mongolian grasslands.

Materials and methods

We sampled soils from inside (ungrazed) and outside (grazed) exclosures in three vegetation types: forest-steppe, shrub-steppe, and desert-steppe. We measured laboratory rates of soil net nitrogen (N) mineralization and net nitrification and geostatistically analyzed heterogeneity.

Results and discussion

Average rates of net N mineralization and net nitrification were lower at shrub-steppe and desert-steppe and were decreased by grazing. Semivariograms showed vegetation-induced heterogeneity in ungrazed plots, except for net nitrification at forest-steppe. We found linear change with distance under dense and uniform vegetation at forest-steppe, 1.3 m patch under patchy vegetation at shrub-steppe, and linear change, but with much smaller semivariance, under sparse and poor vegetation at desert-steppe. At forest-steppe, grazing randomized the spatial patterns of net N mineralization and net nitrification. At shrub-steppe and desert-steppe, grazing greatly decreased the semivariances of net N mineralization and net nitrification as well as their averages, and the soil heterogeneity was virtually disappeared.

Conclusions

Grazing in Mongolian grasslands homogenized the spatial patterns of net N mineralization and net nitrification, irrespective of their original spatial patterns determined by the differences in vegetation structure.     

Extraction and analysis of the tangential arrangement of cambial cells in Japanese hardwood species

Arrangement of the cambium and marginal zone of xylem were illustrated by drawing the outlines of fusi-form cells and tissue on serial tangential sections inZelkova serrata andRobinia pseudoacacia with storied cambium and inAlbizia julibrissin with non-storied cambium. The comparison of cambium drawings with parenchyma drawings revealed that the lengths and arrangement of parenchyma strands correspond closely with those of corresponding fusiform initials in all three species with storied or non-storied cambium. Based on this result, we deduced the correlation between length and variation of length and arrangement of the fusiform initials in 20 species of Japanese hardwoods using parenchyma drawings instead of cambium drawings. The species with a smaller variation of length tend to exhibit a storied arrangement regardless of the length of the fusiform initial. Thus, it is suggested that the variation of the length of the fusiform initial has more influence than the length itself on the development of the storied arrangement.     

Relationship between the incidence of pine wilt disease and the drainage area

We examined the relationship between the incidence of pine wilt disease and moisture conditions in the stand level ofPinus densiflora Sieb. et Zucc. forests in the warm-temperate zone of the western part of Japan. For this analysis, pine trees killed by pine wilt disease were distinguished from the suppressed trees by their position in the layer of the pine forest stand. The drainage area, which is small in the upper part and large in the lower part of the slope, was adopted for representing the moisture conditions in the soil of the slope. The percentage of the pine trees killed by pine wilt disease increased as the size of the drainage area increased. This result suggested that the incidence of pine wilt disease tended to be high in areas with moist conditions. Pine trees attacked by the pinewood nematode die from extensive water deficit due to tracheid cavitations. Pine wilt disease mainly emerges in the summer when the soil water conditions become especially severe, and the radical water stress is thought to accelerate the disease. It was assumed that pine trees in the plots with the small drainage area resisted the influence of the attack of the pinewood nematode because pine trees in the plots with the small drainage area encountered long-term water stress and acquired drought tolerance. Pine trees in the plots with the large drainage area were presumed to be well established in the moist conditions and not to have acquired drought tolerance. The drought tolerance of pine trees was thought to be an important factor in resistance to pine wilt disease.     

Isolation and characterization of a novel anionic peroxidase cDNA found in poplar (Populus nigra) suspension cultured cells

A cDNA clone fromPopulus nigra L. var.italica Koehne, denoted PCY2-6, coding for an anionic peroxidase has been isolated, cloned, and characterized. PCY2-6 is 1160 bp long; and its deduced product, PnC26, contains 343 amino acid residues. The mature protein has a calculated isoelectric point of 4.09. The protein contains two motifs typical of peroxidase and 10 potentialN-glycosylation sites. PnC26 is therefore classified as an anionic peroxidase. The mRNA of the PCY2-6 gene family was detected in immature and mature leaves and in two parts of current-year stems: the shoot tip and the older stem. The mRNA of PCY2-6 gene family was found to localize in the phloem and cortex of the current-year stems. We therefore conclude that expression of the PCY2-6 gene family is related to bark development.     

Distribution of radioactive cesium and stable cesium in cattle kept on a highly contaminated area of Fukushima nuclear accident

Radioactivity inspection of slaughtered cattle is generally conducted using a portion of the neck muscle; however, there is limited information about the distribution of radioactive cesium in cattle. In this study, therefore, we measured not only radioactive cesium but also stable cesium in various tissues of 19 cattle that had been kept in the area highly contaminated by the Fukushima nuclear accident. Skeletal muscles showed approximately 1.5‐3.0 times higher concentration of radioactive cesium than internal organs. Radioactive cesium concentration in the tenderloin and top round was about 1.2 times as high as that in the neck muscle. The kidney showed the highest concentration of radioactive cesium among internal organs, whereas the liver was lowest. Radioactive cesium concentration in the blood was about 8% of that in the neck muscle. Characteristics of stable cesium distribution were almost the same as those of radioactive cesium. Correlation coefficient between radioactive cesium and stable cesium in tissues of individual cattle was 0.981 ± 0.012. When a suspicious level near 100 Bq/kg is detected in the neck of slaughtered cattle, re‐inspection should be conducted using a different region of muscle, for example top round, to prevent marketing of beef that violates the Food Sanitation Act.     

Analysis of volatile compounds released during the grinding of roasted coffee beans using solid-phase microextraction

A dynamic solid-phase microextraction (SPME) method to sample fresh headspace volatile compounds released during the grinding of roasted coffee beans was described and the analytical results using gas chromatography/mass spectrometry (GC/MS) and GC/olfactometry (GC/O) were compared to those of the conventional static SPME sampling methods using ground coffee. Volatile compounds released during the grinding of roasted coffee beans (150 g) were obtained by exposing the SPME fiber (poly(dimethylsiloxane)/divinylbenzene, PDMS/ DVB) for 8 min to nitrogen gas (600 mL/min) discharged from a glass vessel in which the electronic coffee grinder was enclosed. Identification and characterization of volatile compounds thus obtained were achieved by GC/MS and GC/O. Peak areas of 47 typical coffee volatile compounds, separated on total ion chromatogram (TIC), obtained by the dynamic SPME method, showed coefficients of variation less than 5% (n = 3) and the gas chromatographic profile of volatile compounds thus obtained was similar to that of the solvent extract of ground coffee, except for highly volatile compounds such as 4-hydroxy-2,5-dimethyl-3(2H)-furanone and 4-ethenyl-2-methoxyphenol. Also, SPME dilution analysis of volatile compounds released during the grinding of roasted coffee beans showed linear plots of peak area versus exposed fiber length (R (2) > 0.89). Compared with those of the headspace volatile compounds of ground coffee using GC/MS and GC/O, the volatile compounds generated during the grinding of roasted coffee beans were rich in nutty- and smoke-roast aromas.