An effective and practical method of net settings in rearing tank to suppress hypermelanosis in Japanese flounder

In Japanese flounder aquaculture, hypermelanosis occurs widely on the blind side. Rearing flounders in a net-lined tank was recently reported to prevent hypermelanosis. To effectively apply this method to larger tanks for aquaculture farming, the net setting method was examined. Juvenile flounders without darkened areas on the blind side [total length (TL) 13 cm] were selected, and reared for 6 months (TL 32 cm). In the control tank without a net, the median value of darkened area ratio (darkened area to blind side area) was 46%. By only covering the tank bottom with slack net, the darkened area ratio was suppressed to 8%, less than 1/5 of that in the control tank. At the end of experiment, bottom coverage ratios of 0%, 10%, 30%, 50%, and 100% revealed a negative correlation between bottom net coverage and darkened area ratio. In this experiment, the darkening area in the tank with 50% bottom net coverage decreased to 1/3 of the control. Although the occurrence of hypermelanosis differs depending on the production lot, these results are expected to serve as a reference for selecting the suitable net size to meet the level of clearness of the blind side.

     

Semen Collection and Polymerase Chain Reaction‐Based Sex Determination of Black‐Headed and Straw‐Necked Ibis

This study aimed to develop a polymerase chain reaction (PCR)‐based sexing and effective semen collection methods for black‐headed and straw‐necked ibis species. However, most birds are not sexually dimorphic, that is, the sexes appear similar. Therefore, the gender should be determined before semen collection. DNA was extracted from the blood samples of 11 black‐headed and 4 straw‐necked ibis. The sex was determined after PCR amplification of the EE0.6 region of W‐chromosome. The PCR products were separated using gel electrophoresis. A single band indicated the presence of the EE0.6 region and that the individual was a female, while no band indicated that the individual was a male. Further, the single bands from seven specimens were amplified. Semen collection was performed by massage or a combination of massage with electro‐ejaculation and was attempted during all four seasons. The semen was successfully collected in March from male straw‐necked ibis using the massage method. Limited motility, viability and concentration of straw‐necked ibis sperm were observed. The sperm length was 180 μm and that of the nucleus was 30 μm with acrosome located at the tip of the nucleus. Thus, the PCR‐based sexing proved to be an accurate molecular sexing method for black‐headed and straw‐necked ibis. Furthermore, we successfully collected semen and observed the stained sperm nucleus and acrosome of the straw‐necked ibis sperm. We propose that the use of this PCR methodology can be applied as a routine method for sex determination and semen collection in ibis species for future ecological research. However, considering our limited success, further studies on semen collection method are required.     

Loss of virulence of the phytopathogen Ralstonia solanacearum through infection by φRSM filamentous phages

φRSM1 and φRSM3 (φRSM phages) are filamentous phages (inoviruses) that infect Ralstonia solanacearum, the causative agent of bacterial wilt. Infection by φRSM phages causes several cultural and physiological changes to host cells, especially loss of virulence. In this study, we characterized changes related to the virulence in φRSM3-infected cells, including (i) reduced twitching motility and reduced amounts of type IV pili (Tfp), (ii) lower levels of β-1,4-endoglucanase (Egl) activity and extracellular polysaccharides (EPS) production, and (iii) reduced expression of certain genes (egl, pehC, phcA, phcB, pilT, and hrpB). The significantly lower levels of phcA and phcB expression in φRSM3-infected cells suggested that functional PhcA was insufficient to activate many virulence genes. Tomato plants injected with φRSM3-infected cells of different R. solanacearum strains did not show wilting symptoms. The virulence and virulence factors were restored when φRSM3-encoded orf15, the gene for a putative repressor-like protein, was disrupted. Expression levels of phcA as well as other virulence-related genes in φRSM3-ΔORF15-infected cells were comparable with those in wild-type cells, suggesting that orf15 of φRSM3 may repress phcA and, consequently, result in loss of virulence.     

An immunohistochemical study of endocrine cells in the abomasum of vagotomized calf

The effect of thoraco-vagotomy on the distribution and frequency of chromogranin-, serotonin-, somatostatin- and gastrin-immunoreactive cells in the abomasum of the calf were investigated by immunohistochemistry. Calves were vagotomized at 1 week old and sampled 2 and 4 weeks later. The endocrine cells generally decreased in number in vagotomized calves as compared to non-operated control calves. However, the detailed responses of endocrine cells to vagotomy varied depending on the endocrine cell type, region of gastric mucosa, and period after vagotomy. The present result suggests that the vagus nerve has an influence on the intrinsic regulatory system by endocrine cell control in the ruminant abomasum.     

Superoxide generation in extracts from isolated plant cell walls is regulated by fungal signal molecules

ABSTRACT Fractions solubilized with NaCl from cell walls of pea and cowpea plants catalyzed the formation of blue formazan from nitroblue tetrazolium. Because superoxide dismutase decreased formazan production by over 90%, superoxide anion (O(2) ) may participate in the formation of formazan in the solubilized cell wall fractions. The formazan formation in the fractions solubilized from pea and cowpea cell walls was markedly reduced by exclusion of NAD(P)H, manganese ion, or p-coumaric acid from the reaction mixture. The formazan formation was severely inhibited by salicylhydroxamic acid and catalase, but not by imidazole, pyridine, quinacrine, and diphenyleneiodonium. An elicitor preparation from the pea pathogen Mycosphaerella pinodes enhanced the activities of formazan formation nonspecifically in both pea and cowpea fractions. The suppressor preparation from M. pinodes inhibited the activity in the pea fraction in the presence or absence of the elicitor. In the cowpea fraction, however, the suppressor did not inhibit the elicitor-enhanced activity, and the suppressor alone stimulated formazan formation. These results indicated that O(2) generation in the fractions solubilized from pea and cowpea cell walls seems to be catalyzed by cell wall-bound peroxidase(s) and that the plant cell walls alone are able to respond to the elicitor non-specifically and to the suppressor in a species-specific manner, suggesting the plant cell walls may play an important role in determination of plant-fungal pathogen specificity.     

Protective effect of floor cover against soil erosion on steep slopes forested with <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> (hinoki) and other species

 We evaluated the protective effects of floor cover against soil erosion in three types of forest located on steep slopes under a humid climate: 22- and 34-year-old Chamaecyparis obtusa (hinoki), 34-year-old Cryptomeria japonica (sugi), and 62-year-old Pinus densiflora (red pine) stands. We measured sediment transport rates (sediment mass passing through one meter of contour width per millimeter of rainfall), using sediment traps, before and after removing floor cover. Raindrop splash erosion was dominant in the experimental stands. Floor cover percentage (FCP) during the preremoval stage varied from 50% to 100% among the four stands, and sediment transport rates ranged from 0.0079 to 1.7 g m⁻¹ mm⁻¹. The rates increased to 1.5–5.6 g m⁻¹ mm⁻¹ immediately after removing floor cover, and remained high throughout the experiment. The presence of physical cover near the ground has a crucial effect on sediment transport on forested slopes. The protective effect ratio (the ratio of the sediment transport rate in a control plot to that in the removal plot) in a young hinoki stand, in which the FCP decreased markedly, was 0.3 at most, which is close to the rate for bare ground. The protective effect ratio in the red pine stand was ≤0.003. We concluded that the protective effect of floor cover in undisturbed forests in Japan differs by over two orders of magnitude, based on comparisons with previous studies. Received: March 11, 2002 / Accepted: August 16, 2002 Present address: Department of Forest Site Environment, Forestry and Forest Products Research Institute, Ibaraki 305-8687 Japan Tel. +81-298-73-3211; Fax +81-298-74-3720 e-mail: miura@affrc.go.jp Present address: Department of Forest Site Environment, Forestry and Forest Products Research Institute, Ibaraki 305-8687 Japan Tel. +81-298-73-3211; Fax +81-298-74-3720 e-mail: miura@affrc.go.jp Acknowledgments This study was supported by the Research Council of the Ministry of Agriculture, Forestry, and Fisheries, of Japan. We thank H. Ujihara, S. Ujihara, and M. Ogasawara in Otoyo, Kochi, who provided the experimental stands used in this study. We also thank K. Hirai, S. Kuramoto, E. Kodani, and the rest of the staff at the Shikoku Research Center, Forestry and Forest Products Research Institute, for their help in conducting the experiments. Correspondence to:S. Miura     

Methane emissions from stems of Fraxinus mandshurica var. japonica trees in a floodplain forest

We measured methane (CH₄) emissions from the stem surfaces of mature Fraxinus mandshurica var. japonica trees in a floodplain forest. Flux measurements were conducted almost monthly from May to October 2005, and positive CH₄ fluxes were detected throughout the study period, including the leafless season. The mean CH₄ flux was 176 and 97 μg CH₄ m⁻² h⁻¹ at the lower (15 cm above the ground) and upper (70 cm above the ground) stem positions, respectively. The CH₄ concentration was lower in soil gas than in ambient air to a depth of at least 40 cm. One possible source of CH₄ emitted from the stems might be the dissolved CH₄ in groundwater; maximum concentrations were 10,000 times higher than atmospheric CH₄ concentrations. Our results suggest that CH₄ transport from the submerged soil layer to the atmosphere may occur through internal air spaces in tree bodies.     

Antiallergic tea catechin, (-)-epigallocatechin-3-O-(3-O-methyl)-gallate,suppresses FcepsilonRI expression in human basophilic KU812 cells

We previously found that the O-methylated derivative of (-)-epigallocatechin-3-O-gallate (EGCg), (-)-epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG' '3Me), has potent antiallergic activity. The high-affinity IgE receptor, FcepsilonRI, is found at high levels on basophils and mast cells and plays a key role in a series of acute and chronic human allergic reactions. To understand the mechanism of action for the antiallergic EGCG' '3Me, the effect of EGCG' '3Me on the cell surface expression of FcepsilonRI in human basophilic KU812 cells was examined. Flow cytometric analysis showed that EGCG' '3Me was able to decrease the cell surface expression of FcepsilonRI. Moreover, immunoblot analysis revealed that total cellular expression of the FcepsilonRI alpha chain decreased upon treatment with EGCG' '3Me. FcepsilonRI is a tetrameric structure comprising one alpha chain, one beta chain, and two gamma chains. The level of mRNA production of each subunit in KU812 cells was investigated. EGCG' '3Me reduced FcepsilonRI alpha and gamma mRNA levels. The cross-linkage of FcepsilonRI causes the activation of basophils, which leads to the secretion of inflammatory mediators including histamine. EGCG' '3Me treatment inhibited the FcepsilonRI cross-linking-induced histamine release. These results suggested that EGCG' '3Me can negatively regulate basophil activation through the suppression of FcepsilonRI expression.