Resinous stem canker development during the growing season of Chamaecyparis obtusa (Hinoki cypress) inoculated with pathogenic fungus Cistella japonica

Seasonal development of resinous stem canker was determined in Chamaecyparis obtusa trees wound‐inoculated with Cistella japonica in January 1996. Samples for anatomical analysis were harvested on 20 May, 9 July and 6 August in the second year of inoculation, and the sections were observed under light microscope. Resin exudation was most abundant in the second year, decreasing in the third. All new resin exudation in the second year was recognized at the May observation. New traumatic resin canal formation was observed in the August samples, and an expansion of necrotic lesions was observed mainly in summer samples in the inoculated trees and in naturally affected trees. Cistella japonica was re‐isolated from all bark lesions or inoculated wounds examined. These results suggest that the activity of Ci. japonica in the tree affects the seasonal development of resinous stem canker during the growing season of the trees.     

Radical scavenging and singlet oxygen quenching activity of marine carotenoid fucoxanthin and its metabolites

Antioxidant activity of carotenoids is suggested to be one of the factors for their disease preventing effects. Marine carotenoids fucoxanthin and its two metabolites, fucoxanthinol and halocynthiaxanthin, have been shown to exhibit several biological effects. The antioxidant activities of these three carotenoids were assessed in vitro with respect to radical scavenging and singlet oxygen quenching abilities. The 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of fucoxanthin and fucoxanthinol was higher than that of halocynthiaxanthin, with the effective concentration for 50% scavenging (EC 50) being 164.60, 153.78, and 826.39 microM, respectively. 2,2'-Azinobis-3-ethylbenzo thizoline-6-sulphonate radical scavenging activity of fucoxanthinol (EC 50, 2.49 microM) was stronger than that of fucoxanthin (EC 50, 8.94 microM). Hydroxyl radical scavenging activity as measured by the chemiluminescence technique showed that the scavenging activity of fucoxanthin was 7.9 times higher than that by fucoxanthinol, 16.3 times higher than that by halocynthiaxanthin, and 13.5 times higher than that by alpha-tocopherol. A similar trend was observed when the hydroxyl radical scavenging was assessed by the electron spin resonance (ESR) technique. ESR analysis of the superoxide radical scavenging activity also showed the superiority of fucoxanthin over the other two carotenoids tested. Singlet oxygen quenching ability of the three carotenoids was lower than that of beta-carotene, with quenching rate constants ( k Q, x10 (10) M (-1) s (-1)) being 1.19, 1.81, 0.80, and 12.78 for fucoxanthin, fucoxanthinol, halocynthiaxanthin, and beta-carotene, respectively. The higher radical scavenging activity of fucoxanthin and fucoxanthinol compared with halocynthiaxanthin is assumed to be due to presence of the allenic bond.     

Development of a monoclonal antibody-based sandwich ELISA for detection of guinea pig interleukin-2

Interleukin 2 (IL-2) is a T cell proliferation factor released by Th0- and Th1-type helper T cells and is an essential cytokine for immune responses. In the present study, recombinant glutathione S-transferase (GST)-guinea pig IL-2 (GPIL-2) fusion protein was prepared by Escherichia coli (E. coli) and by using this protein as an immunogen, monoclonal antibodies (mAbs) against GPIL-2 were produced to establish a basis for a research on immune responses in guinea pigs. Three stable hybridoma cell lines were established, and specific binding of each mAb to recombinant GPIL-2 produced by E. coli and insect cells infected with recombinant baculovirus was shown by enzyme linked immunosorbent assay (ELISA) and/or immunoblot analysis. Isotype analyses of these mAbs revealed that all three mAbs were IgG1 and had kappa chain. Furthermore, assessment of their epitopes by competition binding assay indicated that the mAbs obtained in this study bound to three different epitopes. Thus, a sandwich ELISA based on the two mAbs specific to different GPIL-2 epitopes was developed for detection of GPIL-2, which had a sensitivity threshold of about 0.3 ng/ml of GPIL-2.     

Lymphography of the thoracic duct by percutaneous injection of iohexol into the popliteal lymph node of dogs: experimental study and clinical application

OBJECTIVE: To evaluate the efficacy of percutaneous administration of iohexol into the popliteal lymph node as a non-invasive technique for thoracic duct lymphangiography in dogs. STUDY DESIGN: Experimental study and clinical report. ANIMALS: Normal adult dogs (n=4) and 1 dog with recurrent chylothorax. METHODS: For the experimental study, 4 dogs (weight, 8.4-12.3 kg) had 5-10 mL iohexol injected percutaneously into 1 popliteal lymph node and then thoracic radiographs were taken. Popliteal lymph nodes were examined by histopathology 8 days later. One 25-kg dog with recurrent chylothorax had 25 mL iohexol injected into the right popliteal lymph node followed by thoracic radiography. RESULTS: In experimental dogs, the thoracic duct was best visualized on thoracic radiographs after administration of 10 mL iohexol. Clinically, no abnormalities were identified in the injected limb and except for 1 dog that had large numbers of siderocytes and erythrophagocytic macrophages in the injected lymph node, the histopathologic findings in the other injected popliteal lymph nodes were not different from contralateral nodes. In the clinical case, the thoracic duct was visualized, but there was leakage of iohexol around the node. CONCLUSION: The thoracic duct in dogs can be visualized by lymphography after percutaneous injection of iohexol (1 mL/kg at 2 mL/min) into the popliteal lymph node. CLINICAL RELEVANCE: Percutaneous popliteal lymph node administration of iohexol should be considered as an alternative to mesenteric lymph node injection for radiographic identification of the thoracic duct in dogs.     

Antioxidant mechanism studies on ferulic acid: identification of oxidative coupling products from methyl ferulate and linoleate

In our studies of the chain-breaking antioxidant mechanism of natural phenols in food components, ferulic acid, a phenolic acid widely distributed in edible plants, especially grain, was investigated. The radical oxidation reaction of a large amount of ethyl linoleate in the presence of the methyl ester of ferulic acid produced four types of peroxides as radical termination products. The isolation and structure determination of the peroxides revealed that they had tricyclic structures which consisted of ethyl linoleate, methyl ferulate, and molecular oxygen. Based on the formation pathway of the products, a radical scavenging reaction occurred at the 3'-position of the ferulate radical with the four types of peroxyl radicals of ethyl linoleate. The produced peroxides subsequently underwent intramolecular Diels-Alder reaction to afford stable tricyclic peroxides.     

Analyses of leaves from open field-grown transgenic poplars overexpressing xyloglucanase

The transgenic expression of Aspergillus xyloglucanase cDNA (AaXEG2) with 35S promoter in the leaves of open field-grown poplars was studied. The level of xyloglucan in the transgenic poplars was decreased to 15–16% in the non-fertile soil (forest-field soil) and to 21–22% in the fertile soil (farming-field soil) compared with that of the wild-type poplars. The leaves exhibited a smaller surface area with more rounded teeth than those of the wild-type plants, similar to the sun leaf variety that was grown in the incubation room and subsequently greenhoused. The majority of total veins with water-conducting vascular bundles were shorter in the leaves of the transgenic poplars than those of the wild type. This decrease in vein length may result from a decrease in xyloglucan during leaf development, from which large numbers of proteins were markedly downregulated in the leaves of the transgenic plants via proteomic analysis. It seems likely that the leaves of the transgenic poplars came to relax the edges of their tooth rather than extend their veins as a result of the loosening of the xyloglucan cellulose networks in the leaves.     

Effect of polymorphism in egg white lysozyme on muramidase and antibacterial activities as well as hatchability in the Japanese quail (Coturnix japonica)

Lysozyme is one of the best characterized antimicrobial proteins in egg white. Three phenotypes of egg white lysozyme in Japanese quail, Coturnix japonica, (namely fast; slow; and the combination, FS) were observed by acid polyacrylamide gel electrophoresis. The fast phenotype showed faster mobility on Acid-PAGE than the slow phenotype. Comparison of the coding sequences for lysozyme derived from the slow and fast phenotypes revealed a nonsynonymous SNP at nucleotide position 115 from the translation initiation site, which alters AA sequence of lysozyme. This nonsynonymous SNP converted glutamine (Q) in the slow phenotype to lysine (K) in the fast phenotype at AA residue 21 of mature lysozyme (Q21K). Here, we investigated the effect of these phenotypes on muramidase activity, antibacterial activity, and hatchability. Muramidase activity toward isolated cell walls of Micrococcus lysodeikticus was in the order: fast allozyme > slow allozyme > chicken (Gallus gallus), but no significant difference was found among the 3 (P > 0.05). Antibacterial activity against live Staphylococcus aureus cells was significantly greater for the fast allozyme than the slow allozyme from 20 h after incubation (P < 0.05). For the antibacterial effects against live Escherichia coli cells, the activity of fast was significantly higher than that of slow at 16 h after incubation (P < 0.05). Hatchability was estimated for reciprocal crosses of Japanese quail with the FF (fast) and SS (slow) genotypes. Hatchability was 92.5% in FF male × SS female crosses and 87.2% in SS male × FF female crosses. A Cochran-Mantel-Haenszel test revealed a significant difference between the crosses (P < 0.05) and indicated that the female-derived slow phenotype led to improved rates of hatching. Our results suggest that the nonsynonymous SNP in Japanese quail lysozyme influences the electrophoretic migration, muramidase activity, and antibacterial activity of the protein, in addition to the hatchability of the eggs. These results demonstrate, for the first time, a significant difference in antibacterial activity and hatchability between 2 lysozyme phenotypes in Japanese quail.     

Comparison of feed intake, digestibility and fattening performance of Brahman grade cattle (Bos indicus) and crossbred water buffalo (Bubalus bubalis)

Twenty growing crossbred cattle and crossbred water buffalo (carabao) with an average age of 22 (18–24 months) months were equally distributed into two treatment groups according to species. The animals were fed with the same ration made up of corn silage (50%) + wet brewer's spent grain (30%) + concentrate mixture (20%), and their fattening performance was monitored. The digestibilities of the different nutrients were likewise determined. The economics of raising the animals under intensive production system was calculated. Species differences did not influence total dry matter intake of the animals, when expressed as percentage of the bodyweight and per metabolic body size. There were no significant differences in digestion coefficients of the different nutrients, except for crude protein in crossbred water buffalo and crossbred cattle, although the digestibility of dry matter, organic matter and nitrogen free extract tended to be high in the former than in the latter. Likewise, average daily gain (ADG) was similar, although crossbred water buffalo had numerically higher ADG (828.6 vs 785.5 g) than crossbred cattle during the 6 months feeding. During the first 3 months of feeding (1–90 days), the ADG of crossbred water buffalo was 1066.1 g compared to 940.1 g for crossbred cattle. From 91 to 180 days, the crossbred cattle had slightly higher ADG (630.1 vs 591.1 g) but also the difference was not significant. The return above feed cost was comparable for crossbred cattle and crossbred water buffalo during the first 90 days of feeding. However, extending the feeding period from 91 to 180 days , income over feed cost was higher (P < 0.05) for crossbred cattle by PhP 5.3/kg gain than crossbred water buffalo. Results showed that crossbred water buffalo could attain similar growth rate with that of crossbred cattle under intensive system, when fed with high quality feed materials.     

Density and sound-speed contrasts,and target strength of Japanese sandeel <Emphasis Type="Italic">Ammodytes personatus</Emphasis>

Sound-speed and density contrasts (h and g, respectively), important acoustic material properties, of Japanese sandeel Ammodytes personatus were measured to estimate theoretical target strength (TS). The measured sound-speed contrast of adult fish varied between 1.016 and 1.023 (mean 1.020), and showed temperature dependence. The measured density contrast differed significantly between juvenile and adult. The density contrast of juvenile varied between 1.017 and 1.024 (1.021), and that of adult varied between 1.026 and 1.038 (1.032). Using these results, TS at 38 and 120 kHz in the fishing season were estimated by an empirical sound scattering model. TS of an individual fish varied significantly with change of tilt angle. TS of near-dorsal aspect (TS_max) and tilt-averaged TS (TS_ave) differed by up to 7 dB. At both frequencies, two different TS_ave−length relationships (TS_ave = a log L + b) were obtained for adult and juvenile. The coefficients of log L of adult were close to 20, suggesting that backscattering strength was proportional to square of body length. These values were larger in juvenile (34.0 at 120 kHz, 56.5 at 38 kHz), suggesting that backscattering strength varied drastically with the cube or fifth power of body length.     

Toward sustainable management of small cetacean fisheries around Japan

Abstract:   The management of Japanese domestic fisheries for small cetaceans has been based on a traditional approach that involves only the best set of assumptions and data. However, uncertainty, which is pervasive and inevitable for most fisheries resources, often leads the traditional approach to serious failure. Here is presented the basic framework of a simulation-based approach with a simple Bayesian method that is applicable to Japanese fisheries for small cetaceans. For illustration, the simulation model was applied to Dall's porpoise data in order to investigate the robustness of several management procedures against uncertainty. The simulation showed that the current management procedure, based on only the best values, could fail to manage the stocks at a considerably high probability. However, the use of the conservative management procedure, potential biological removal, permits the sustainable harvesting of Dall's porpoise for at least 100 years into the future, even under uncertainty. In conjunction with a firm structure for implementation of management procedures, the spread of simulation-based approaches will quickly enable successful sustainable management of small cetaceans.