Demonstration of the absence of intervening sequences (IVSs) within 16S rRNA genes of Taylorella equigenitalis and Taylorella asinigenitalis isolates

A total of 57 Taylorella equigenitalis (n=22) and Taylorella asinigenitalis (n=35) isolates was shown not to carry any intervening sequences (IVSs) within 16S rRNA gene sequences. By contrast, we have already shown the genus Taylorella group to carry several kinds of IVSs within the 23S rRNA gene sequences.     

3-Methylthiopropionic acid ethyl ester, isolated from Katsura-uri (Japanese pickling melon, Cucumis melo var. conomon), enhanced differentiation in human colon cancer cells

The fully ripened fruit of Katsura-uri Japanese pickling melon ( Cucumis melo var. conomon) has rarely been used for food because the midripened fruit is utilized for making pickles, but the fully ripened fruit is no longer valuable for pickles due to the fruit body being too soft. We have considered the utilization of the fully ripened Katsura-uri fruit that may be used for nonpickling products, particularly if the fully ripened fruit demonstrated health benefits such as anticarcinogenic properties. The phytochemical extract from the fully ripened fruit of Katsura-uri Japanese pickling melon was purified via a bioassay-guided fractionation scheme, which was based on the induction of differentiation in a RCM-1 human colon cancer cell line. On the criteria of two differentiation markers (duct formation and alkaline phosphatase activity), the most potent fraction contained a compound identified as 3-methylthiopropionic acid ethyl ester, based on GC retention time, EI-MS, (1)H NMR, and (13)C NMR spectra. Previously, the role of 3-methylthiopropionic acid ethyl ester was considered as an odor producing compound in many fruits, but this study indicates potential medical benefits of this compound.     

Production of transgenic cloned pigs expressing the far-red fluorescent protein monomeric Plum

Monomeric Plum (Plum), a far-red fluorescent protein with photostability and photopermeability, is potentially suitable for in vivo imaging and detection of fluorescence in body tissues. The aim of this study was to generate transgenic cloned pigs exhibiting systemic expression of Plum using somatic cell nuclear transfer (SCNT) technology. Nuclear donor cells for SCNT were obtained by introducing a Plum-expression vector driven by a combination of the cytomegalovirus early enhancer and chicken beta-actin promoter into porcine fetal fibroblasts (PFFs). The cleavage and blastocyst formation rates of reconstructed SCNT embryos were 81.0% (34/42) and 78.6% (33/42), respectively. At 36–37 days of gestation, three fetuses systemically expressing Plum were obtained from one recipient to which 103 SCNT embryos were transferred (3/103, 2.9%). For generation of offspring expressing Plum, rejuvenated PFFs were established from one cloned fetus and used as nuclear donor cells. Four cloned offspring and one stillborn cloned offspring were produced from one recipient to which 117 SCNT embryos were transferred (5/117, 4.3%). All offspring exhibited high levels of Plum fluorescence in blood cells, such as lymphocytes, monocytes and granulocytes. In addition, the skin, heart, kidney, pancreas, liver and spleen also exhibited Plum expression. These observations demonstrated that transfer of the Plum gene did not interfere with the development of porcine SCNT embryos and resulted in the successful generation of transgenic cloned pigs that systemically expressed Plum. This is the first report of the generation and characterization of transgenic cloned pigs expressing the far-red fluorescent protein Plum.     

Fatal winter dysentery with severe anemia in an adult cow

An adult dairy cow fatally affected with winter dysentery was investigated pathologically and virologically. The cow had severe anemia and diarrhea with massive blood. Pathologically, the loss of surface epithelial cells and necrosis of crypt epithelial cells in the large intestine were observed. Bovine coronavirus (BCV) antigen was observed in necrotic crypt epithelial cells of the large intestine. Virus particles were found in the necrotic epithelial cells of the large intestine. Virologically, BCV was isolated from the feces of the dead cow. The dead cow had no serum antibody against BCV although the co-habitants did. These suggest that severe infection of BCV in the cow without the BCV antibody accompanied by severe hemorrhagic anemia resulted in the cow's death.     

Malachite-green-removing properties of a bacterial strain isolated from fish ponds in Thailand

Malachite green (MG) has been focused on as a biotreatment target and its biological properties have also been an issue in food fish aquaculture. An MG-removing bacterium was isolated from aquaculture fish pond sediment samples in Thailand. The isolate, strain T-5-2, is a Gram-negative, aerobic rod-shaped bacterium, and has been identified as a member of the Pseudomonas putida group. Proton nuclear magnetic resonance spectroscopy (¹H-NMR) analysis of a broth culture medium containing MG showed that the concentration of MG decreased markedly and that other molecules, including leucomalachite green (LMG), were generated. Moreover, liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis showed that the MG concentration in the broth culture medium continuously decreased. This analysis also demonstrated that the concentration of LMG initially increased and then gradually decreased. Furthermore, gas chromatography–mass spectrometry (GC–MS) analysis showed 4-(dimethylamino)benzophenone (4DABP) as a degradation component of MG, which was confirmed by ¹H-NMR and LC–MS/MS analysis. These findings suggest that this bacterial strain can remove MG in broth culture and degrade it to certain metabolites, including LMG and 4DABP. This study is the first detailed evaluation by the combination of LC–MS/MS, GC–MS, and ¹H-NMR analyses of an MG-removing bacterium isolated from Thai aquaculture fish ponds.     

Adverse and beneficial effects of long-term high-concentration ascorbic acid supplementation in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

The present study examined the effects of high-concentration ascorbic acid (AsA) supplementation over a long period on growth, plasma components, nonspecific immune responses, and thermal tolerance in rainbow trout Oncorhynchus mykiss. Four commercial diets supplemented with 0, 100, 1000, and 5000 mg AsA per kg of diet (designated AsA0, AsA100, AsA1000, and AsA5000, respectively) were fed to rainbow trout (initial weight 1.85 g) for 100 days. AsA contents in liver increased with increasing dietary AsA levels. Feeding period, growth performance, and plasma components did not differ significantly between the AsA groups. On the other hand, the phagocytic assay [nitro-blue tetrazolium (NBT) assay] was significantly higher in fish fed AsA1000 and AsA5000 than those fed a diet without AsA (AsA0) at the end of feeding trial. Superoxide dismutase (SOD) activity in the intestines of fish fed AsA5000 was also significantly higher than that of fish fed AsA0. In the thermal tolerance test, fish fed AsA5000 only showed significantly lower cumulative mortality compared with the AsA0 group. In conclusion, high-concentration AsA supplementation such as AsA5000 over a long period does not induce any adverse effects and can enhance disease and stress resistance in rainbow trout.     

Phylogeography of the Japanese eight-barbel loach <Emphasis Type="Italic">Lefua echigonia</Emphasis> from the Yamagata area of the Tohoku district,Japan

The Japanese eight-barbel loach Lefua echigonia, which is a freshwater fish native to Japan, is distributed from the Tohoku to Kinki districts and is divided into six regional populations according to mtDNA analysis. In this study, we investigated L. echigonia collected from several locations in Yamagata Prefecture and neighboring prefectures using mtDNA control region sequences and confirmed the spatial distribution pattern among the new regional population (Yamagata population). The new population was limited to the Mogami river system in the inland area of Yamagata Prefecture and is distinguished from other regional populations by high sequence divergences.     

The influence of auxin type on the array of somaclonal variants generated from somatic embryogenesis of eggplant, Solanum melongena L.

Somaclonal variation is a possible source of variation in plant breeding. To apply this approach to eggplant breeding, somaclonal variations were observed among plants regenerated through somatic embryogenesis induced by 1-naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). Variations including leaf shape, plant height, flower number per cluster, fruit shape, anther number per flower and pollen fertility were compared among ? 300 plants (R₀). Although these variations were confirmed among plants regenerated using both auxins, the frequencies of somaclonal variations in leaf shape, plant height, fruit shape and pollen fertility in the NAA experiment were higher than those in the 2,4-D experiment. Variations in flower number and anther number were also confirmed among plants from both experiments, although no significant differences in their frequency were observed. Subsequently, the inheritance of variations (leaf shape, fruit shape, and flower number) observed in R₀ was investigated from generation to generation (R₁). Variations in leaf shape and fruit shape were inherited while those in flower number were not. From these results we concluded that, although a high frequency of somaclonal variations were observed among plants from both methods, embryogenesis with NAA was more efficient than 2,4-D in eggplant.     

Seasonal variations in major components of Crassostrea gigas from Seto Inland Sea

Fisheries Science - The proximate composition and nutritional content of oysters define their quality and commercial value. Cultivation site and harvest date may impact these characteristics. We...