Effect of crushing unhulled rice and the addition of fermented juice of epiphytic lactic acid bacteria on the fermentation quality of whole crop rice silage, and its digestibility and rumen fermentation status in sheep

This experiment was carried out to examine the effect of crushing unhulled rice and the addition of fermented juice of epiphytic lactic acid bacteria (FJLB) on the fermentation quality of whole crop rice silage, and its digestibility and rumen fermentation in sheep. After whole rice plants (Oryza sativa L. cv. Haenuki) were harvested between the yellow and full‐ripened period of maturity, and cut at a length of 2 cm, four silage treatments were set: (i) non‐treatment; (ii) crushing treatment of unhulled rice division (crushing); (iii) addition of 1% FJLB; and (iv) crushing + addition of 1% FJLB. Lactic acid content increased with the addition of FJLB, and was highest in the crushing + FJLB treatment. Digestibilities of crude fiber, acid detergent fiber and neutral detergent fiber were significantly higher during the crushing + FJLB treatment. There were no differences among treatments in the volatile fatty acid concentrations before and after feeding. At 4 h after feeding, however, the ratio of acetic acid : propionic acid was significantly lower with the addition of FJLB. It was proven that the fermented quality of silage was improved when FJLB was added to rice plants and then ensilaged, and the digestibility of the fibrous component was raised when unhulled rice division was crushed and FJLB was supplemented.     

The relationship between changes in serum glycine and alanine concentrations in non-essential amino acid and milk production in the transition period in dairy cows

The serum concentration of non-essential amino acid (NEAA) was measured in ten Holstein dairy cows grouped as low production (n=5) and high production (n=5) from one month pre-partum through four months post-partum and the relationship between production and amino acid concentrations was studied. The glycine (Gly)/NEAA ratio and the glycine/alanine ratio of the high production group were significantly higher than the low production group (p<0.01). The observed decrease of the alanine (Ala)/NEAA ratio was more remarkable in the high production group than in the low production group. Measurement of Gly/Ala ratio in serum may be useful for evaluating the nutritional status of peri-parturient dairy cow.     

Excretion patterns of fecal progestagens, androgen and estrogens during pregnancy, parturition and postpartum in okapi (Okapia johnstoni)

The aim of the present study was to establish a simple method to monitor ovarian activity and non-invasively diagnose pregnancy in okapi (Okapia johnstoni). The feces of a female okapi were collected daily or every 3 days for 28 months. Steroids in lyophilized feces were extracted with 80% methanol, and the fecal levels of immunoreactive progestagens (progesterone and pregnanediol-glucuronide), androgen (testosterone), and estrogens (estradiol-17beta and estrone) were determined by enzyme immunoassays with commercially available antisera. Using the progesterone profiles, the durations of the luteal phase, follicular phase, and estrous cycle were determined to be 11.1 +/- 0.4, 5.3 +/- 0.6, and 16.5 +/- 0.7 days (n=22), respectively. Fecal levels of immunoreactive progesterone, pregnanediol glucuronide, and testosterone gradually increased from early pregnancy and peaked several months before parturition. More pregnanediol glucuronide was excreted in feces than progesterone during late pregnancy, but not during the estrous cycle. Although the fecal concentrations of immunoreactive estradiol-17beta and estrone change a little throughout pregnancy and non-pregnancy, they rose sharply and temporarily on the day following parturition. The present study indicates that fecal assays with commercial antisera for progesterone and pregnanediol glucuronide are useful for evaluating luteal activity and diagnosing pregnancy and indicates that estrogens might have some role as a trigger of parturition.     

Kujigamberol, a new dinorlabdane diterpenoid isolated from 85million years old Kuji amber using a biotechnological assay

A new compound, 15,20-dinor-5,7,9-labdatriene-18-ol (1), named kujigamberol, was isolated from amber, fossilized tree resin from the Kuji area in Japan, has been dated as being 85 million years old (late Cretaceous). Kujigamberol was identified using the hypersensitive mutant yeast (zds1? erg3? pdr1? pdr3?) with respect to Ca²⁺-signal transduction. The structure was elucidated on the basis of spectroscopic analysis including 1D NMR, 2D NMR and HR-EI-MS. It was different from known diterpenoids with a similar activity isolated from Baltic amber (agathic acid 15-monomethyl ester (2), dehydroabietic acid (3) and pimaric acid (4)). Kujigamberol showed glycogen synthase kinase-3β (GSK-3β) inhibition activity involving the growth restored activity against the mutant yeast and was cytotoxic to HL60 cells (IC₅₀ = 19.6 μM).     

Determination of FVIIa-sTF Inhibitors in Toxic Microcystis Cyanobacteria by LC-MS Technique

The blood coagulation cascade involves the human coagulation factors thrombin and an activated factor VII (fVIIa). Thrombin and fVIIa are vitamin-K-dependent clotting factors associated with bleeding, bleeding complications and disorders. Thrombin and fVIIa cause excessive bleeding when treated with vitamin-K antagonists. In this research, we explored different strains of toxic Microcystis aeruginosa and cyanobacteria blooms for the probable fVIIa-soluble Tissue Factor (fVIIa-sTF) inhibitors. The algal cells were subjected to acidification, and reverse phase (ODS) chromatography-solid phase extraction eluted by water to 100% MeOH with 20%-MeOH increments except for M. aeruginosa NIES-89, from the National Institute for Environmental Studies (NIES), which was eluted with 5%-MeOH increments as an isolation procedure to separate aeruginosins 89A and B from co-eluting microcystins. The 40%–80% MeOH fractions of the cyanobacterial extract are active against fVIIa-sTF. The fVIIa-sTF active fractions from cultured cyanobacteria and cyanobacteria blooms were subjected to liquid chromatography-mass spectrometry (LC-MS). The 60% MeOH fraction of M. aeruginosa K139 exhibited an m/z 603 [M + H]⁺ attributed to aeruginosin K139, and the 40% MeOH fraction of M. aeruginosa NIES-89 displayed ions with m/z 617 [M − SO₃ + H]⁺ and m/z [M + H]⁺ 717, which attributed to aeruginosin 89. Aeruginosins 102A/B and 298A/B were also observed from other toxic strains of M. aeruginosa with positive fVIIa-sTF inhibitory activity. The active fractions contained cyanobacterial peptides of the aeruginosin class as fVIIa-sTF inhibitors detected by LC-MS.     

Introduction of a fungal NADP(H)-dependent glutamate dehydrogenase (gdhA) improves growth,grain weight and salt resistance by enhancing the nitrogen uptake efficiency in forage rice

We report the effect of the introduction of a fungal glutamate dehydrogenase gene (gdhA) into forage rice (cv. Momiroman) by analyzing the transgenic rice plants in terms of growth, source function, and nitrogen contents. NADP (H)-dependent glutamate dehydrogenase activities in the gdhA-transgenic lines were markedly higher than those in non-transgenic control plants. Plant growth analysis at the seedling stage revealed that the leaf area and shoot and root dry weights of the high gdhA-expressors were higher than those of control plants under both high (high N) and low nitrogen (low N) conditions. These results suggested that the source ability was enhanced by the gdhA introduction. This was supported by the fact that the net photosynthesis rate at the heading stage was also higher in transgenic than in control leaves. Furthermore, under both high and low N conditions, the nitrogen contents in the shoots and roots, at seedling and grain-harvest stages, were significantly higher in high gdhA-expressors than in control plants, indicating that nitrogen uptake was higher in transgenic than in control plants. At the harvest stage, the high gdhA-expressors exhibited greater panicle and spikelet numbers per plant compared with control plants, resulting in higher grain weight, under the high N conditions. In addition, gdhA expression in forage rice significantly enhanced their tolerance to salt stress compared to control plants. The present study showed that the introduction of a fungal gdhA into forage rice could lead to higher source ability, better growth and higher grain weight by enhancing nitrogen uptake efficiency.     

Inflammatory effect of cleaved bovine lactoferrin by elastase on staphylococcal mastitis

Elastase activity and concanavalin A (Con A) low affinity bovine lactoferrin (bLf) molecule were detected in mammary gland secretions (MGSs) from mammary glands (MGs) with clinical staphylococcal mastitis. Changes in clinical symptoms correlated with increases in both elastase activity and the concentration of Con A low-affinity Lf in MGSs from mastitic MGs. Bovine Lf treated with elastase (elastase-Lf) showed various small bLf molecules and the same image on Con A two-dimensional immunoelectrophoresis as low Con A affinity bLf in MGSs. We confirmed the presence of four common bLf peptides for the elastase-bLf and low Con A affinity bLf molecules in mastitic MGSs, and synthesized four peptides. Strong mRNA expression of interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1) was induced in bovine mammary epithelial cells on stimulation with low Con A affinity bLf, elastase-bLf, and GQRDLLFKDSAL, a synthesis bLf peptide based on nuclear factor kappa B (NFkappaB) activation. These results suggest that bLf was cleaved by elastase, and that this cleavage changed the physical function of Lf. Our results indicate that elastase induced production of low Con A affinity bLf, including the bLf peptide GQRDLLFKDSAL, and had an inflammatory effect on staphylococcal mastitis.     

Bait twig method for soil detection of <Emphasis Type="Italic">Rosellinia necatrix</Emphasis>, causal agent of white root rot of Japanese pear and apple,at an early stage of tree infection

Mulberry twigs were inserted into the soil as bait to detect Rosellinia necatrix at an early stage of tree infection in the orchard. R. necatrix was frequently trapped on twigs near the trunk base at soil depths of 6–20 cm within 10–20 days in May–July, suggesting that the incubation period was dependent on soil temperature. Subsequently, we inserted twig in the soil around healthy-looking trees in naturally infested orchards. R. necatrix was trapped from 80.0% of Japanese pear and 75.0% of apple trees that later proved to be infected. This bait twig method facilitated quicker diagnosis of white root rot on Japanese pear and apple at early stages of infection and can be used to detect recurrence of the fungus after fungicide treatment.