Latex agglutination test: a simple, rapid and practical method for bovine serum CRP determination.

A semi-quantitative latex agglutination test for bovine serum CRP levels has been established by mixing diluted serum (or diluted standard serum) with a 1% latex suspension containing 0.489 micron latex particles coated with affinity-purified antibody at a ratio of 20 micrograms/mg latex. The agglutination was performed on a glass slide in a moist chamber at room temperature with 45 min. incubation. This test is reliable, reproducible and the results correlate with those of the single radial immunodiffusion (SRID) test. The effect of low temperature storage on CRP concentration revealed a 30% degradation of CRP during 2 years storage at 4 degrees C. The possible role of EDTA addition to prevent a decrease in serum CRP concentration by freezing and thawing is also discussed.     

Changes in concentrations of plasma metabolites and insulin-like growth factor-1 after feeding in Japanese Black and Japanese Shorthorn cattle

The aim was to characterize the changes of plasma component concentrations after feeding in Japanese Black (B) and Japanese Shorthorn (S) cattle. Eight females, five B and three S, with a mean bodyweight (standard deviation) of 282.0 (20.3) kg were selected and supplied with only water for 28 h after feeding. Blood was taken from the jugular vein at 4‐h intervals for 32 h after feeding. Metabolite (cholesterol, CL; glucose, GLU; nonesterified fatty acid, NEFA; triacylglycerol, TG; total protein, TP) and insulin‐like growth factor‐1 (IGF‐1) concentrations in plasma were measured. In both breeds GLU tended to decrease linearly with a similar slope. Differences between the initial mean NEFA at 4 h after feeding (Start) and that at 24 h after the start (End) were significant in both breeds, but S increased significantly more than B at and after 8 h from the Start. The TG in S tended to change more than in B, and CL and IGF‐1 in both breeds changed continuously at similar levels. The TP in B tended to decrease more slowly and be lower than S. The change in NEFA, especially, suggests that nutritional status sensitively reflects genetic background.     

Susceptibility of various animals and cultured cells to exfoliative toxin produced by Staphylococcus hyicus subsp. hyicus

In piglets inoculated with partially purified exfoliative toxin (pp-shET) produced by Staphylococcus hyicus subsp. hyicus, exfoliation was observed at 12 h after injection. Chickens inoculated with the same dose of pp-shET also showed exfoliation within 30 min of injection. However, exfoliation was not demonstrated in mouse, rat, guinea pig, hamster, dog or cat inoculated with pp-shET until 24 h after injection. In cultured cell lines, especially L-929 and Hep-2, the rounding effect occurred after incubation with pp-shET for 1 h. The rounding effect was also seen in five other cultured cells (NCTC 2544, HeLa/S3, HmLu-1, CHO and BHK-21) 6-24 h after exposure to pp-shET. These round cells survived for 72 h after inoculation and formed a monolayer 24 h after changeover to a toxin-free medium. The rounding effect was observed in cells after the formation of the monolayer, but not before. It was suggested that the rounding effect was not caused by the increase in cyclic AMP in cells inoculated with pp-shET but by the cleavage of intracellular contacts.     

Isolation of exfoliative toxin from Staphylococcus hyicus subsp. hyicus and its exfoliative activity in the piglet

Exfoliative toxin was isolated from the sterile cell-free filtrate of 24 h culture of Staphylococcus hyicus subsp. hyicus strain P-1. The partial purification of exfoliative toxin produced by S. hyicus (shET) was performed by precipitation with 50-80% saturated ammonium sulfate, gel filtration on a Sephadex G-75 column and column chromatography on DEAE-cellulose. Partially purified shET (pp-shET) caused exfoliation in piglets at 8 to 12 h after intradermal or subcutaneous injection. However, heat-treated pp-shET did not cause exfoliation in piglets for up to 24 h after injection. On histopathological examination of the skin at 12 h after injection of pp-shET, an intraepidermal cleavage plane was shown between the stratum corneum and stratum granulosum and at the stratum granulosum.     

Insulin administration suppresses an increase in insulin-like growth factor binding protein-2 gene expression stimulated by fasting in the chicken

1. We examined the changes in plasma IGF-I concentration and tissue IGFBP-2 gene expression of young fasted chickens refed a commercial diet or administered bovine insulin intravenously. 2. Plasma IGF-I concentration was decreased by fasting for 2 d. Although plasma IGF-I concentration was increased by refeeding, it didn't recover to the level of chickens fed a commercial diet ad libitum. 3. Insulin administration lowered plasma IGF-I concentration compared to other groups. 4. Hepatic IGFBP-2 mRNA was increased by fasting for 2 d and decreased by refeeding for 6 h. Insulin administration also decreased hepatic IGFBP-2 gene expression stimulated by fasting to the level of refed chickens. 5. IGFBP-2 mRNA in the gizzard was increased by fasting for 2 d and tended to decrease after refeeding for 6 h. Insulin administration decreased gizzard IGFBP-2 gene expression to less than that in refed chickens. 6. There was no between-treatment difference in IGFBP-2 mRNA in the brain and kidney. 7. These results suggest that the changes in IGFBP-2 gene expression in the liver and gizzard by fasting and refeeding might be partly regulated by the alteration in plasma insulin concentration.     

A study of reproductive performance in pregnant, IL-2 receptor beta-chain overexpressed transgenic mice

Relationships between female reproductive performance and uterine natural killer (uNK) cells were investigated in pregnant IL-2 receptor beta-chain overexpressed transgenic (Tg2Rbeta) mice. At 8 days of pregnancy, all fetuses were alive, suggesting that implantation normally occurred in these mice. However, 47% of fetuses were dead at 10 days of pregnancy and at 12 days all fetuses were resorbing, indicating that fetal loss progressed with the advance of pregnancy. The placenta of Tg2Rbeta mice gradually decreased in weight with the advance of pregnancy. At 10 days the placental labyrinth, decidua basalis, and metrial gland in Tg2Rbeta mice were poorly developed, and more uNK cells were found in Tg2Rbeta mice than in the control mice. We propose that Tg2RPbeta mice are the first and interesting model that uNK cells can cause abortion, to clarify the involvement of uNK cell function in female reproductive performance.     

Rumen fermentation,milk production and conjugated linoleic acid in the milk of cows fed high fiber diets added with dried distillers grains with solubles

The effects of corn dried distillers grains with solubles (DDGS) feeding on rumen fermentation and milk production in cows were evaluated using diets high in neutral detergent fiber (NDF, 45.9–46.6%). The control diet (Control) consisted mainly of hay, corn silage and concentrates. In the experimental diets, the concentrates were replaced with DDGS as 10% dry matter (DM) (10%DDGS) and 20% DM (20%DDGS). Eight cows were used for each 14‐day treatment period. Effect of DDGS feeding on DM intake was not significant. Ruminal volatile fatty acids and ammonia‐N at 5 h after feeding of 20%DDGS were decreased compared to Control, whereas protozoal count at 2 h after feeding of 20%DDGS was higher than that of 10%DDGS. Milk yield of cows fed DDGS diets was greater than that of Control, although percentages of milk protein and solids‐not‐fat were decreased by DDGS diets. The proportions of C10:0, C12:0, C14:0 and C16:0 in the milk fat decreased, and those of C18:0, C18:1, C18:2 and cis‐9, trans‐11 conjugated linoleic acid (CLA) increased markedly with elevated DDGS. Increase in trans‐11 C18:1 was observed in the rumen fluid at 5 h after feeding. These findings suggest that DDGS feeding enhanced milk yield, as well as CLA synthesis under a high dietary NDF condition.     

The assessment of genetic diversity within and among the eight subpopulations of Japanese Black cattle using 52 microsatellite markers

Japanese Black cattle are at risk for genetic homogeneity due to intensive use of a few sires. Therefore, assessment of the actual genetic diversity of this breed is important for future breeding plans. In the present study, we investigated the genetic diversity within and among eight subpopulations of Japanese Black cattle using 52 microsatellite markers. The parameters for genetic diversity of Japanese Black cattle were comparable to those of other cattle breeds, suggesting that the relatively high genetic diversity of the breed. However, upon comparison among the eight subpopulations, the Hyogo subpopulation showed markedly low genetic diversity. The results of the pairwise F_ST values, phylogenetic network and structure analysis indicated that the Hyogo population has remarkably high level of genetic differentiation from other populations, while Yamagata, Niigata, Hiroshima and Kagawa populations have low levels of genetic differentiation. Furthermore, multidimensional scaling plots indicated that individuals in some subpopulations were separated from individuals in the other subpopulations. We conclude that while the overall genetic diversity of Japanese Black cattle is still maintained at a relatively high level, that of a particular subpopulation is significantly reduced, and therefore the effective population size of the breed needs to be controlled by correct mating strategies.