Effect of mate selection on fuzzy selective mating criteria in closed dairy multiple ovulation and embryo transfer nucleus programs

In order to control rates of response and inbreeding, mate selection using fuzzy selective mating criteria (FMC) was investigated in adult multiple ovulation and embryo transfer nucleus schemes for dairy cattle. Stochastic simulation was used to model the closed nucleus scheme. This mate selection was examined in four alternative mating and male selection schemes: (i) a hierarchical scheme; (ii) a hierarchical sibship scheme (two males per sibship); (iii) a factorial scheme (two sires per dam); and (iv) a factorial sibship scheme (two males per sibship and two sires per dam). Genetic response and inbreeding rate tended to be reduced by increasing the trade-off parameter of FMC between the expected breeding value and inbreeding of progeny. Inbreeding rates in all schemes were reduced by reducing the variance of family size through selection and the average coancestry of mating pairs through mate allocation.     

Involvement of a host erythrocyte sialic acid content in Babesia bovis infection

Host sialic acid (SA) has recently been suggested to play an important role in erythrocyte (RBC) infection by Babesia spp. The present study attempted to further determine the specific type of SAs important in the RBC invasion. Bovine RBC was found to bear abundant alpha2-3-linked SA residues but not alpha2-6-linked SA in nature, confirmed by flow cytometric analysis of the neuraminidase (Nm)-treated RBCs. Lectin-blot analyses revealed the removal of alpha2-3-linked SAs from the 97-, 33-, and 31-kDa bands by the Nm treatment. Addition of the Nm-treated RBCs into an in vitro culture of B. bovis resulted in a decreased population of the parasitized RBCs. The thin smear samples from the cultures were then observed under a confocal laser scanning microscope after staining with the alpha2-3-linked SA-specific lectin: a selective invasion of B. bovis was found only in the intact RBCs bearing the SAs, but not in the desialylated RBCs. Furthermore, a significant reduction of the parasitized RBCs was also observed in the culture supplemented with exogenous 3'-sialyllactose containing the alpha2-3-linked SAs. However, the complete inhibition of parasite proliferation was not achieved in the culture. These findings indicate that while the alpha2-3-linked SA-dependent pathway is needed for highly efficient invasion of host RBCs by B. bovis, there might also be other potential alternative pathways.     

Spindle formation and microtubule organization during first division in reconstructed rat embryos produced by somatic cell nuclear transfer

The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 microM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-alpha-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization.     

Comparison of flower color with anthocyanin composition patterns in evergreen azalea

In evergreen azaleas, major anthocyanins were detected from petals of wild species and cultivars by HPLC analysis. Depending on flower color, all samples were divided into three groups: red, purple or white, using the Japan color standard for horticultural plants. The chromatic components a* and b* values of red group samples showed a convergent distribution, whereas those of purple group samples showed a wider distribution. According to the HPLC analysis, red group samples had two to four major anthocyanins, and those of the purple group had two to six major ones. In contrast, no anthocyanins were detected in the white group petals, although anthocyanidins were detected. These results suggest that the anthocyanin constitution of the purple group flowers is more varied than that of the red group flowers, and this wider variety among purple flowers contributes to extending the diversity of flower color in evergreen azalea.     

Varietal differences in phenolic content and astringency in skin and flesh of hardy kiwifruit resources in Japan

Varietal differences in the total phenolic content and astringency in the skin and flesh were determined among the cultivars and local collections of hardy kiwifruit with a ploidy variance found in Japan. The average values of the total phenolic content in the skin and flesh were 2.66 and 0.18 g 100 g⁻¹ FW, respectively. There were large varietal differences in the total phenolic content in the skin in the range of 1.3–5.0 g 100 g⁻¹ FW. Kochi (tetraploid), while Gassan and ‘Mitsuko’ (hexaploid) contained a larger amount of total phenolics. High astringency was found in Gassan, ‘Mitsuko’ and ‘Hoko’ (hexaploids) and Kochi. HPLC analysis showed that the major components of phenolics in the flesh were (+)-catechin, chlorogenic acid, rutin, (−)-epicatechin and quercetin.     

Estimation of daily urinary potassium excretion using urinary creatinine as an index substance in prepartum dairy cows

In the present study, the daily excretion of potassium (K) in urine (urinary K(UK)) was estimated from a 6 h urine sample using urinary creatinine (UC) as the index substance. All urine was collected from six pregnant Holstein cows at 6 h intervals for 24 h on 3 days of the 4th, 2nd and final week before the expected date of parturition. In total, 72 6 h urine samples were obtained. Daily UC excretion (mg/day per kg bodyweight (BW)) was almost the same for the three sampling days. Daily UC excretion varied among cows from 22.1 to 24.3 mg/day per kg BW with a mean of 22.8 mg/day per kg BW with no significant difference. Thus, daily UC excretion was confirmed to be constant throughout the prepartum period with no differences among individuals. The concentration ratios of K to creatinine ((UK mg/dL)/(UC mg/dL) (UK/UC)) correlated strongly to the hourly K excretions (mg/h per kg BW) (r = 0.952, P < 0.01) in the 6 h urine samples. The differences in the UK/UC ratio between sampling periods were not significant within each cow. Therefore, daily UK excretion (mg/day) can be estimated using the equation: daily UK excretion (mg/day) = daily UC excretion (mg/day per kg BW) × BW (kg) × 6 h urine sample UK/UC, where daily UC excretion can be a given value.     

Phylogenetic Analysis of Sugarcane Rusts Based on Sequences of ITS, 5.8 S rDNA and D1/D2 Regions of LSU rDNA

Phylogenetic analysis of sugarcane rusts based on sequences of ITS and the 5.8 S rDNA revealed two highly divergent ITS groups among isolates of Puccinia sp. sensu Muta, 1987 and P. kuehnii specimens. Although there is sufficient divergence (exceeding normal intraspecific variation) between the ITS regions of the two groups to support separation into different species, unusually high homology of the ITS group I sequences with those of members of Cronartium and identical sequences of the D1/D2 regions of the LSU rDNA for all the isolates of “Puccinia sp.” and P. kuehnii that otherwise exhibited different ITS sequences, suggest that the two highly divergent sequences may have resulted from abnormal genetic events leading to non-orthologous, intraspeciflc polymorphisms. The other sugarcane rust, P. melanocephala and the grass rusts, P. miscanthi and P. rufipes, were separated from “Puccinia sp.” and P. kuehnii and from each other in D1/D2 region analyses, indicating that D1/D2 region sequences may more correctly reflect phylogenetic relationships in these rusts than do the ITS regions. Further studies to examine differences in patho-genicity or finer morphological features within P. kuehnii that may be correlated with the high divergence in ITS sequences and experiments to determine if these two sequence types represent intraspeciflc polymorphism are necessary. Received 11 October 2000/ Accepted in revised form 24 November 2000     

An Improved Method for Isolating Violet Root Rot Fungus, Helicobasidium mompa, from Basidiocarps

Basidiocarps of the violet root rot fungus, Helicobasidium mompa, are less frequently used for isolation than are mycelial strands and sclerotia even though the basidiocarps are conspicuously produced at the trunk base of diseased plants. Basidiocarps are also more suitable for storage. This paper describes an improved method for obtaining pure cultures from basidiocarps using microcentrifuge tubes to facilitate the awkward steps of rinsing fungal materials under a dissecting microscope. Received 28 June 2000/ Accepted in revised form 4 August 2000