The effect of replacing brewers' grains with barley tea grounds in total mixed ration silage on feed intake, digestibility and ruminal fermentation in wethers

Four wethers were used in a 4 × 4 Latin square to study the feed intake, apparent digestibility, nitrogen balance and ruminal fermentation characteristics when fed total mixed ration (TMR) silages which included wet barley tea grounds (WBTG). The TMR silages were prepared using compound feed including wet brewers' grains (WBG), corn, oat hay, alfalfa hay, dried beet pulp, salt and vitamin-mineral supplement in a ratio of 30.7:15:8:24:10:12:0.15:0.15, respectively, on a dry matter (DM) basis. The WBTG and soybean meal mixture (7:3 on DM basis) were substituted for WBG at ratio of 0% (Control), 5% (LTG), 10% (MTG), and 15% (HTG) on DM of TMR. WBTG addition to the TMR silages increased lactic acid concentration, decreased pH, acetic acid and ammonia-N ( P  < 0.001). Feed intakes and digestibilities for LTG and MTG (except ether extract (EE) digestibility) treatments were not different from the control ( P  > 0.05). However, EE and neutral detergent fiber (NDF) intake, crude protein, EE and NDF digestibility was lower, but the DM and gross energy digestibility was higher for the HTG treatment compared to control ( P  < 0.01). With progressive increases in WBTG concentrations, nitrogen intake, fecal nitrogen and retention nitrogen did not differ, but the urinary nitrogen for MTG and HTG treatments were lower than that of the control ( P  = 0.001). The ruminal total volatile fatty acid concentration and the molar ratios of propionate and butyrate were higher, but the acetate, ratio of acetate to propionate and ammonia-N content were lower for the HTG treatment compared with the control ( P  < 0.05). Therefore, the possible proportion of replacing WBG with WBTG for TMR silage can be 10% or less of the diet DM.     

ISSR variations in eggplant (Solanum melongena L.) and related Solanum species

Inter-simple sequence repeat (ISSR) analysis was performed in eight cultivars of eggplant (Solanum melongena) and 12 accessions in eight related Solanum species to evaluate the applicability of this analysis for assessing the phylogenetic relationships and identifying cultivars. A total of 552 polymorphic amplified bands were obtained from 34 of the 100 primers tested, and the percentage of polymorphisms was 99.1%. Cluster analysis based on the ISSR markers classified the Solanum species into seven groups: (i) S. melongena; (ii) S. aethiopicum and S. anguivi; (iii) S. incanum; (iv) S. violaceum and S. kurzii; (v) S. macrocarpon; (vi) S. virginianum and (vii) S. torvum. Combining the ISSR markers obtained by a few of the 34 primers was enough for distinguishing of the eight cultivars of eggplant. This ISSR analysis was demonstrated to be available for the phylogenetic study and the cultivar identification.     

The mechanism by which mouse spermatozoa are injured during freezing

To improve the cryopreservation protocol for mouse sperm, we attempted to estimate the type and extent of cryoinjury at various steps of the process. First, we demonstrated that mouse sperm are sensitive to chilling at -15 C and that the sensitivity is dependent on the length of exposure. To estimate cryoinjuries, sperm suspensions were ice-seeded at -5 or -15 C, frozen with liquid nitrogen (LN(2)) gas and then frozen in LN(2). In one experiment, sperm seeded at -5 C were cooled slowly to -15 C before deep freezing. At various steps of the cryopreservation process, the sperm were warmed and their viability was assessed based on motility and the integrities of the plasma membrane and acrosome. The motility of frozen-thawed sperm was higher on seeding at -5 C (28%) than at -15 C (9%). The motility did not decrease when the sample was transferred from LN(2) gas to LN(2). To estimate cryoinjury of sperm, we presumed the viability of frozen sperm to be decreased by chilling, hypertonic stress and formation of intracellular ice. When the sperm suspension was cooled and seeded at -5 C, the motility decreased by 25% due to hypertonic stress. When the sperm were cooled in LN(2) gas, the motility decreased by 17% with the formation of intracellular ice. When the sperm were cooled to -15 C, the motility decreased by 51% from chilling. After seeding, the motility decreased by 18% due to formation of intracellular ice and by 7% due to hypertonic stress. Considering the results, it would be preferable to seed samples at a higher temperature to prevent intracellular ice from forming and to cool seeded samples rapidly enough to minimize chilling injury and hypertonic stress, but not too rapidly to allow intracellular ice to form.     

Birth of normal offspring from mouse eggs activated by a phospholipase Czeta protein lacking three EF-hand domains

Sperm-specific phospholipase C, PLCzeta, is a candidate for the Ca(2+) oscillation-inducing factor that is introduced into the ooplasm upon sperm-egg fusion. In addition to the 647-residue full-length PLCzeta, s-PLCzeta lacking the N-terminal 110 amino acids is known to be present in the mouse testis. In this study, we attempted to obtain full-term offspring from s-PLCzeta-activated eggs by round spermatid injection. Metaphase II-arrested eggs injected with a high RNA concentration of s-PLCzeta RNA normally developed to blastocysts. When the round spermatid nucleus was injected into telophase II-stage eggs previously activated by s-PLCzeta RNA, three live offspring were successfully obtained by transfer of the developed 4-cell embryos to pseudopregnant mice. These three offspring all grew to be normal adults and reproduced healthy second-generation mice.     

Molecular evidence of the hybrid origins of leyland cypress (×Cupressocyparis leylandii)

Leyland cypress (×Cupressocyparis leylandii) has been regarded as an intergeneric hybrid between Monterey cypress (Cupressus macrocarpa) and Alaska cypress (Chamaecyparis nootkatensis) in the Cupressaceae. Each partial sequence of the 18S-rDNA from nuclear and therbcL from chloroplast DNA genomes was determined using above three species. At the 59th site from the 5′ end of the 152 bp of the determined 18S-rDNA sequence, one base difference was identified between Monterey cypress (T) and Alaska cypress (A). However, Leyland cypress showed either nucleotide A or T at this site. Since nuclear DNA is inherited by biparental mode, this result shows genetic evidence that the nucleic genome of Leyland cypress originates from Monterey cypress and Alaska cypress and that this species is of hybrid origin.     

Evaluation of antixenosis in soybean against Spodoptera litura by dual-choice assay aided by a statistical analysis model: Discovery of a novel antixenosis in Peking

The method for evaluating soybean (Glycine max) antixenosis against the common cutworm (Spodoptera litura) was developed based on a dual-choice assay aided by a statistical analysis model. This model was constructed from the results of a dual-choice assay in which Enrei, a soybean cultivar susceptible to S. litura, was used as both a standard and a test leaf disc for 2nd–5th instar larvae. The statistical criterion created by this model enabled the evaluation of the presence of antixenosis. This method was applied to four soybean varieties, including Tamahomare (susceptible), Himeshirazu (resistant), IAC100 (resistant), and Peking (unknown), as well as Enrei. Subsequently, the degrees of antixenosis were also compared by F-test, followed by maximum likelihood estimation (MLE). According to the results, the antixenosis of Tamahomare, Himeshirazu, and IAC100 was statistically reevaluated and Peking exhibited a novel antixenosis, which was stronger for 3rd–5th instar larvae than for 2nd instar.     

Composition and Regiodistribution of Fatty Acids in Triacylglycerols and Phospholipids from Red and Black Rices

Regiospecific profiles of fatty acids (FA) of triacylglycerols (TAG) and phospholipids (PL) isolated from red and black rices were investigated. The lipids extracted from red and black rices were separated by thin‐layer chromatography (TLC) into eight subfractions, respectively. With a few exceptions, the major lipid components were TAG (76.4–80.5%), free FA (7.2–9.8%), and phospholipids (3.5–3.6%), while hydrocarbons, steryl esters, diacylglycerols (1,3‐DAG and 1,2‐DAG), and monoacylglycerols were present in minor proportions (0.1–4.1%). The PL components isolated from the two cultivars were phosphatidyl choline (52.3–53.7%), phosphatidyl ethanolamine (22.3–23.1%), phosphatidyl inositol (20.6–21.3%), and others (<3.4%). No significant differences (P < 0.05) in FA distribution were found when these cultivars were compared. The principal characteristics of the FA distribution in the TAG and PL were evident in the rices between the two cultivars: unsaturated FA were predominantly located at the sn‐2 position (77.3–96.8%), while saturated FA primary occupied the sn‐1 or the sn‐3 position (35.0–78.7%) in these lipids. The results of this study could provide useful information to both consumers and producers during manufacture of traditional rice foods in Japan.     

Effect of sulfonamide antibiotics on microbial diversity and activity in a Californian Mollic Haploxeralf

Purpose  

Up to 90% of antibiotics that are fed to livestock are excreted unaltered or as metabolites and thus are present in manure. By application of manure as fertilizer, veterinary antibiotics can reach soil and groundwater. The aim of this study is to determine the effect of three commonly used (and simultaneously applied) sulfonamide antibiotics on both function and structural diversity of soil microorganisms. To this end, the activity of the enzymes urease and dehydrogenase was determined, and the composition of phospholipid fatty acids (PLFA) was analyzed.     

Effect of gestational exposure to nonylphenol on the development and fertility of mouse offspring

Nonylphenol (NP), a kind of environmental chemical, is thought to imitate endogenous hormones, inhibit the actions of hormones, and induce reproductive abnormalities. A number of experimental animals, usually rats, have been used to evaluate the potential reproductive toxicity of NP. However, the findings of previous studies were contradictory in some cases. Therefore, we used ICR mice as a biomodel for in utero study of NP. After mating, 8- to 12-week-old females were assigned to four groups (n=8) for subcutaneous injections from day 5 to 20 of gestation. Group I animals received corn oil alone as a control, while the mice of groups II, III and IV received NP at concentrations of 1/1000, 1/100 and 1/10 of the LD(50), respectively. A dose-dependent decrease was observed in terminal body weights of males of the F1 generation; however, a very small negative effect was only found in the females of the NP1/10 group. No significant effect was found on the liver weights of both sexes. The weights of the testis and epididymis were slightly decreased in the NP1/10 group. The NP1/100 treatment increased ovary weight considerably. The uterus weight tended to be increased in the NP treatment groups; however, there were large variations. The gestational exposure of the groups had no significant effect on the rate of pregnancy (94.4-100%) and the number of fetuses per litter (13.6-14.3 males, 12.3-13.7 females) compared with the control group. However, the overall mortality of fetuses/embryos was increased considerably in the NP1/100 (male: 13.9%) and NP1/10 (female: 9.8%) groups. These results suggest that exposure to NP in utero possibly affects the body weight and some reproductive organ weights, but does not influence the potential fertility of the F1 generation.     

Analysis of the promoter of mutated human whey acidic protein (WAP) gene

Although whey acidic protein (WAP) has been identified in the milk of a range of species, it has been predicted that WAP is not secreted into human milk as a result of critical point mutations within the coding region. In the present study, we first investigated computationally the promoter region of mutated human WAP genes by comparing with those of other known WAP genes. Computational database analyses showed that the human WAP promoter region was highly conserved, as in other species with milk WAP. Next, we evaluated the activity of the human WAP promoter (2.6 kb) using a reporter gene assay. MCF-7 cells were stably transfected with the hWAP/hGH (human growth hormone) fusion gene, cultured on Matrigel, and treated with lactogenic hormones. Radioimmunoassay detected hGH in the culture medium, indicating that the human WAP promoter was responsible for the lactogenic hormones. The human WAP promoter was significantly more active in MCF-7 cells than the mouse WAP promoter (2.4 kb). The present results provide us with important information on the molecular evolution of milk protein genes.