Effects of slaughter age on the levels of free amino acids and dipeptides in fattening cattle

The effects of slaughter age, breed type and postmortem period were studied on the concentrations of free amino acids (FAA) and dipeptides (carnosine and anserine) in Longissimus dorsi muscle of beef. Slaughter age affected the levels of most FAA and dipeptides, and the concentrations of these compounds were significantly lower at 35 months of age than either 15 or 25 months of age. Slight increases were observed at 25 months compared with 15 months, but the differences were not significant except for a few FAA. These tendencies still remained when the concentrations were recalculated on the basis of protein. A significant breed effect was observed for taurine, carnosine and anserine. Almost all FAA were significantly increased during postmortem conditioning, but the level of each FAA fluctuated, and the fluctuations could not be explained statistically by slaughter age or breed differences.     

Estimation of the base line imagined as a muscular background for myoglobin determination by reflectance spectrometry

Liquid extraction method for determination of myoglobin in muscle is not suitable because of changes in the derivatives and time required. Optical reflectance method is useful as a non‐destructive and real‐time measurement. However, the base line of reflectance method, the muscular reflectance background (MRB) imagined as colorless structural elements, has been unknown because of non‐existence. The spectrum of the base line was analyzed as the analogy of bleached meat. A formula was derived for reflectance that had a linear relationship to the reciprocal of wavelength to the power 4. The reflectance characteristic of the MRB was not constant and the curve tended to rise with increasing wavelength. The reflectance at 700 nm was estimated to be 50%. By the characteristic analysis on Commission Internationale l’Eclairage color values and reciprocal reflectance, it was concluded that the reflectance spectrum of the muscular background obtained in this research was fitted as a base line for the reflectance spectrometry.     

Novel polymorphism of the canine dopamine receptor D4 gene intron II region

Various dog breeds are remarkably different from each other not only in their sizes and shapes but also in behavioral traits, suggesting that some of these characteristics are under genetic control. However, little is known about genes related to behavioral traits in canine species. In humans, it has been reported that the dopamine receptor D4 gene (DRD4) includes polymorphism at several regions that relate to personality or psychiatric disorders. In an earlier study by the authors of the present study, the polymorphisms in canine DRD4 exon III and exon I regions were reported. In the present study, a novel polymorphism in canine DRD4 intron II was found based on a 17 base pair insertion/deletion, and the two alleles detected were named P (shorter allele) and Q (longer allele). The allelic distribution in 28 breeds of dog, including a total of 1114 unrelated individuals, were then investigated. Both P and Q alleles were detected in most of the breeds investigated; however, the frequencies of P and Q differed greatly between breeds. With respect to classification based on breed origin, P and Q alleles were frequent in Occidental and Oriental breeds, respectively. Furthermore, two subspecies of wolves, the ancestors of dogs, were analyzed for the comparison of allele frequencies with dogs, and the P allele was predominant in both European and Chinese wolves.     

Butt rot ofChamaecyparis obtusa (Sieb. et Zucc.) Endlicher trees caused byPerenniporia subacida in Shikoku district, Japan: Pathogen, distribution of damaged trees in the stand, and soil investigation

The rot ofChamaecyparis obtusa (hinoki) trees was studied in the northern part of Kagawa Prefecture, Japan. Among 158 hinoki examined, butt rot was found in 28.6, 58.3, and 100% of trees in the 29, 30, and 34-year-old stands, respectively. All trees with butt rot, and 11 trees without it had rotted roots. Thirteen hinoki were peeled byCervus nippon (sika deer) and all of them were infected with butt rot. The maximum height of rot in deer-damaged trees was 2.6 m from the ground level, but was 1.9 m in undamaged trees. About eighty-two percent of rotted trees showed rotted areas of less than 40 cm² on the cross section of stems at the ground level. White mycelia and black flecks sometimes appeared in the rotted wood. Basidiocarps of polyporaceous fungus were often found on felled logs and rotted stumps of hinoki and identified asPerenniporia subacida. Basidiomycetous fungus was isolated frequently from rotted wood of roots and stems, and determined to beP. subacida by comparative study on cultural characteristics. An inoculation experiment and wood-decay test proved that the fungus was the cause of the rot of hinoki. Few absorbing roots of living trees were found in the clayey subsurface soil of the high dry bulk density and the less soil aeration. Some absorbing roots had root rot and the rot spread from the base of the absorbing root to the central part of the woody root. This is the first report on the rot of hinoki caused byP. subacida in Japan.     

切花月季芽变品种的分子标记与鉴别研究

现代月季的基因型高度杂合，目的基因的定位如同大海捞针，形态指标模棱两可的描述比比皆是。为此，特别收集了组别17个，品种45个及相对的芽变品种进行随机增护多态DNA（Random Amplified Polymorphic DNA,RAPD）基因分析。结果表明：11个经筛选的人工全盛随机引物（10－base）共增扩出基因位点143个，其中多态性位点93个，平均多型性比率高达66.0％；切花月季品种与品种之间，特异性RAPD图带清晰稳定，极易识别；由同一品种变异而来的芽变品种之间，特异性位点极少，相似性遗传距离非常接近，最小为0.8607，最大为1；幼叶DNA双步CTAB提取法有助于提高基因指纹图谱的重现性。这为表型性状较为相近的切花月季芽变品种的分子鉴别提供了可靠的技术手段，也为转基因品种的育成奠定了基础。     

Time-dependent changes in inhibitory action of lipopolysaccharide on intestinal motility in rat

Endotoxin causes gastrointestinal motility disorder. Aim of this study is to clarify inhibitory mechanisms of lipopolysaccharide (LPS) on smooth muscle contraction in rat ileum. Ileal tissues were isolated from control rat or from LPS-induced peritonitis model rat. Treatment with LPS inhibited carbachol (CCh)-mediated contraction in a time-dependent manner. Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) genes were also upregulated, but iNOS expression was preceded by a rising of COX-2. All subtypes of prostaglandin E₂ (PGE₂) receptors (EP1-EP4) were expressed in ileum, and PGE₂ and selective EP2 or EP4 agonist inhibited CCh-mediated contraction. Selective iNOS inhibitor did not reverse LPS-induced inhibition of contraction by CCh at 1 and 2 hr, but reduced the inhibitory action at 4 hr after the LPS treatment. COX-2 inhibitor reversed the inhibitory action by LPS in all exposure time. Finally, in ileal tissues isolated from peritonitis model rat, iNOS expression was upregulated only at 4 hr after LPS administration, resulting in enhanced inhibitory action of LPS against CCh-induced contraction. In conclusion, LPS induces COX-2 to produce PGE₂, which initially activates EP2 and/or EP4 on smooth muscle cells to inhibit the contractility in early phase of LPS exposure. Moreover, in late phase of LPS treatment, iNOS is expressed to produce NO, which in turn inhibited the contraction by CCh. The inhibitory cascade is similar in the ileum isolated from peritonitis model rat, indicating time-dependent changes of inhibitory action by LPS on intestinal motility in peritonitis.     

Analysis of a pair of END+ and END? viruses derived from the same bovine viral diarrhea virus stock reveals the amino acid determinants in Npro responsible for inhibition of type I interferon production

The Exaltation of Newcastle disease virus (END) phenomenon is induced by the inhibition of type I interferon in pestivirus-infected cells in vitro, via proteasomal degradation of cellular interferon regulatory factor (IRF)-3 with the property of the viral autoprotease protein N^pro. Reportedly, the amino acid residues in the zinc-binding TRASH motif of N^pro determine the difference in characteristics between END-phenomenon-positive (END⁺) and END-phenomenon-negative (END⁻) classical swine fever viruses (CSFVs). However, the basic mechanism underlying this function in bovine viral diarrhea virus (BVDV) has not been elucidated from the genomic differences between END⁺ and END⁻ viruses using reverse genetics till date. In the present study, comparison of complete genome sequences of a pair of END⁺ and END⁻ viruses isolated from the same virus stock revealed that there were only four amino acid substitutions (D136G, I2623V, D3148G and D3502Y) between two viruses. Based on these differences, viruses with and without mutations at these positions were generated using reverse genetics. The END assay, measurements of induced type I interferon and IRF-3 detection in cells infected with these viruses revealed that the aspartic acid at position 136 in the zinc-binding TRASH motif of N^pro was required to inhibit the production of type I interferon via the degradation of cellular IRF-3, consistently with CSFV.     

Effect of dietary histidine on contents of carnosine and anserine in muscles of broilers

Carnosine (β‐alanyl‐L‐histidine) and anserine (β‐alanyl‐1‐methyl‐L‐histidine) are dipeptides mainly found in skeletal muscle and brain of many vertebrates, and particularly high concentrations are observed in chicken pectoral muscles. It was reported that these peptides have many functions, such as antioxidant activity. In this study, we examined the effect of different levels of dietary histidine on carnosine and anserine contents in broiler muscles. The 14‐days‐old female Chunky strain broilers were given feeds containing three different levels of histidine; 67% (Low‐His), 100% (Control) and 200% (High‐His) of histidine requirement according to the NRC (1994). Chicks were fed experimental diets for 10 days. Both dipeptides in muscle were significantly decreased. In particular, carnosine was not detected at all in the Low‐His group and was significantly increased in the High‐His group. Both dipeptides were not detected in plasma. These results indicated the possibility to produce chicken meat with enhanced amount of these dipeptides by high histidine feeding.     

Preliminary evaluation of diagnostic tests for avian influenza using the Markov Chain Monte Carlo (MCMC) Method in an emergency surveillance

In June 2005, an outbreak of avian influenza (AI) caused by a low pathogenic H5N2 virus was identified in Japan. A serological surveillance was conducted because the infected chickens did not show any clinical signs. The Markov Chain Monte Carlo Method was used to evaluate the performances of serological HI and AGP tests because there was not enough time when the surveillance was initiated to conduct a test evaluation. The sensitivity of the AGP test (0.67) was lower than that of the HI test (0.99), while the specificities were high for both tests (0.96 for AGP and 0.90 for HI). Based on the low sensitivity of the AGP test, the HI test was used for primary screening in later stages of the epidemic.     

Occurrence of 'Candidatus Mycoplasma turicensis' infection in domestic cats in Japan

We have examined the presence of hemoplasmas, hemotropic mycoplasmas, among domestic cats (Felis catus) in Japan by using a species specific PCR, and found 'Candidatus Mycoplasma turicensis', a recently recognized hemoplasma species. A total of 60 feline blood samples collected in 2004 and 2005 were subjected to PCR amplification for the detection of Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis'. Six blood samples collected from domestic cats were found infected with the 'Candidatus M. turicensis'. All of them are also infected with other species of hemoplasmas, M. haemofelis and/or 'Candidatus M. haemominutum'. This is the first to demonstrate 'Candidatus M. turicensis' infections among cat population in Japan.