Isolation of microsatellite markers by in silico screening implicated for genetic linkage mapping in Japanese pufferfish Takifugu rubripes

ABSTRACT:   The search for dinucleotide repeat microsatellites within scaffolds 1–25 of genome database JGI Fugu v3.0 for the pufferfish Takifugu rubripes revealed that 80% of microsatellite loci consisted of five to 13-fold repeats with locus-specific differences in density. Eleven out of 15 microsatellite loci isolated from the database with which genotyping using wild pufferfish was successfully performed showed polymorphism; that is, the means of the number of alleles and expected and observed heterozygosities at these 11 loci were 21.8, 0.915 and 0.829, respectively. It was confirmed that eight out of the 11 polymorphic loci were inherited through the Mendelian law and one pair of microsatellite loci derived from the same scaffold was linked. These results demonstrated that these loci are useful for constructing a linkage map in the pufferfish as DNA markers.     

A review of ghost fishing: scientific approaches to evaluation and solutions

ABSTRACT:   Research on ghost fishing became active in the late 1980s. Ghost fishing has been confirmed for traps, gillnets, trammel-nets and small seine nets. Some lost traps are functional for a long period of time, even in shallow waters. Consequences for gillnets after loss depend on seabed conditions. The ghost fishing function of gillnets remaining on flat seabeds declines rapidly with decreasing heights and increasing visibility. Gillnets left tangled around an artificial reef, for example, three-dimensionally maintain the initial magnitude of ghost fishing for a long period of time, even after badly fouled. There are increasing numbers of researches working on the total number of mortality per gear after gear loss for gillnets and trammel-nets. It has become also possible to estimate the total number of mortality for a unit period of time in a certain fishing sector. This paper reviews research which has provided evidence and quantitative data on ghost fishing, and proposes five items important for future studies on ghost fishing.     

Inhibitory effects of epigallocatechin gallate on the propagation of bovine coronavirus in Madin-Darby bovine kidney cells

Epigallocatechin gallate (EGCg) is the main active component of tea polyphenol and shows several biological activities, such as antimicrobial, antitumor‐promoting, anti‐inflammatory and anti‐oxidative activities. In the present study, the inhibitory effect of EGCg on bovine coronavirus (BCV) propagation in Madin‐Darby bovine kidney (MDBK) cells was investigated. EGCg at concentrations of less than 10 µg/mL did not show any cytotoxicity to MDBK cells. BCV propagation was significantly inhibited by pretreatment of the virus with EGCg (0.5–10 µg/mL) before virus inoculation in dose‐dependent, incubation time‐dependent and temperature‐dependent manners. The antiviral effect of pretreating MDBK cells with EGCg on BCV propagation was much weaker than that of pretreating BCV with EGCg. The hemagglutination activity of BCV was also reduced by EGCg in a dose‐dependent manner. These results demonstrate that EGCg possesses a distinct anti‐BCV activity and strongly suggest that EGCg interferes with the adsorption of BCV to MDBK cells by the interaction of EGCg with BCV particles. EGCg may therefore be a useful candidate for controlling BCV infection more effectively.     

Effect of methotrexate on cerebellar development in infant rats

Six-day-old rats were treated intraperitoneal injections with methotrexate 1 mg/kg, and the cerebellum was examined. Both the length and width of the vermis decreased in the methotrexate-treated group instead of the control from 4 day after treatment (DAT) onward. A significant reduction in the width of the external granular layer was detected on 2 and 3 DAT in the methotrexate group. By 4 DAT, the width of the external granular layer of the methotrexate group was indistinguishable from the control, and by 8 DAT, it was greater than that of the control. The molecular layer of methotrexate group on 8 and 15 DAT was thinner than that of the control. On 1 DAT, in the methotrexate group, there were many TUNEL and cleaved caspase-3-positive granular cells throughout the external granular layer, and they decreased time-dependently. On 1 DAT, in the methotrexate group, phospho-histone H3-positive cells in the external granular layer were fewer than in the control and tended to increase on 2–4 DAT. The p21-positive-rate of the external granule cells in the MTX group was higher than in the control on 1–4 DAT. These results suggested that methotrexate exposure on postnatal day 6 induces a delay, slowing in the migration of external granular cells to the inner granular layer, attributed to decrease or inhibition in the production of external granular cells that had arisen from apoptosis and the decrease in cell proliferative activity, resulting in cerebellar hypoplasia.     

Black leaf spot and circular leaf spot of <Emphasis Type="Italic">Farfugium japonicum</Emphasis> (L.) Kitamura caused by <Emphasis Type="Italic">Phoma</Emphasis> spp.

In May 1998 and 1999, two types of leaf spot (black type and brown type) caused by Phoma spp. were found on Farfugium japonicum in Tokyo and in Gunma Prefecture, Japan. The fungus isolated from black-type lesions caused only black-type lesions, and the fungus from brown-type lesions caused only brown-type lesions. We propose to name these diseases black leaf spot of F. japonicum (kokuhan-byo in Japanese) for the disease with black lesions and circular leaf spot of F. japonicum (rinmon-byo in Japanese) for the disease with brown lesions. This is the first report on leaf diseases of F. japonicum caused by Phoma spp.     

Botrytis blight of Taiwanese toad lily caused by Botrytis elliptica (Berkeley) Cooke

In June 1998, leaf spots caused by Botrytis elliptica (Berkeley) Cooke were found on Taiwanese toad lily (Tricyrtis formosana Bak.) in Kanagawa Prefecture, Japan. This is the first report on Botrytis blight of T. formosana caused by B. elliptica.     

Leaf spot of <Emphasis Type="Italic">Aster savatieri</Emphasis> (Makino) Kitam. caused by <Emphasis Type="Italic">Septoria chrysanthemella</Emphasis> Saccardo

 In May 1998 leaf spot caused by Septoria chrysanthemella was found on Aster savatieri in Kanagawa Prefecture, Japan. This is the first report of leaf spot on A. savatieri caused by S. chrysanthemella. Received: September 13, 2002 / Accepted: October 18, 2002 Acknowledgments The authors thank Dr. T. Kobayashi, formerly of Tokyo University of Agriculture, for his advice on identifying the fungus.     

Evaporative light-scattering analysis of sulforaphane in broccoli samples: Quality of broccoli products regarding sulforaphane contents

Broccoli sulforaphane has received attention as a possible anticarcinogen. Sulforaphane analysis is difficult due to the lack of a chromophore for spectrometric detection. Hence, we developed a method for determining sulforaphane by using high-performance liquid chromatography (HPLC) coupled with an evaporative light-scattering detector (ELSD). Sulforaphane was extracted from acid-hydrolyzed broccoli samples, followed by solid-phase extraction and reversed-phase HPLC. Sulforaphane was detected by ELSD and concurrently identified by electrospray ionization time-of-flight mass spectrometry. The recovery of sulforaphane from broccoli samples was above 95%. The detection limit was 0.5 mug. The present method was sensitive enough to determine sulforaphane in mature broccoli, broccoli sprouts, and commercial broccoli products. Sulforaphane concentration in broccoli sprout (1153 mg/100 g dry weight) was about 10 times higher than that of mature broccoli (44-171 mg/100 g dry weight). Therefore, the broccoli sprout is recommended as a source of sulforaphane-rich products. In contrast, we found that sulforaphane could not be detected in most of broccoli products, suggesting present commercial broccoli products having low quality.     

Role of Starch Granules in Controlling Expansion of Dough During Baking

The role of starch granules in the expansion of doughs during baking was investigated using artificial flours made from dry vital wheat gluten and wheat starch, potato starch, or tapioca starch. The three starches were selected because of their diverse gelatinization properties. Baking tests on flour from tapioca starch gave the largest loaf volume and the most extensive postbaking shrinkage. Potato starch flour gave the smallest volume and the least shrinkage. Amylograph test data, dough expansion under decreased pressure, progress of expansion during baking, and scanning electron microscopy revealed the role starch granules play in ideal baking conditions. Starch granules should not gelatinize early in the baking cycle as potato starch does but should gelatinize later in the baking cycle as wheat starch does. This prevents early setting of the dough which inhibits expansion. Starch granules should not disrupt and fuse together during gelatinization as tapioca starch does, forming an impermeable gas membrane. Granules should gelatinize individually as wheat starch does, causing a disruption of cell membranes which prevents shrinkage of the loaf during cooling after baking.