Comparison of morphological and DNA-based techniques for stomach content analyses in juvenile chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis>: a case study on diet richness of juvenile fishes

To identify the stomach contents of marine organisms, morphological observations are commonly conducted. However, the results obtained by this traditional method are frequently biased, as it is difficult to detect partially digested, soft-bodied organisms. To resolve this, recent studies have used DNA-based (DNA barcoding) analyses to examine the diet breadth of marine organisms. Here, we compared the prey richness of juvenile fishes using morphological observations and DNA-based analyses, with a focus on juvenile chum salmon Oncorhynchus keta. A higher number of prey taxa were successfully identified using DNA-based analyses than morphological observations. However, we also noticed several shortcomings of the DNA-based analyses, as reported in other diet-analysis studies that used molecular techniques. For example, the degree of digestion among prey taxa might have resulted in differential sensitivity to DNA detection. Additionally, several prey taxa could not be precisely identified, as the sequence data for some of the targeted organismal groups are unavailable in public gene databases. Remarkably, it is also possible that DNA-based analyses detected secondary prey, and therefore, the richness of prey taxa was likely overestimated. Thus, dietary analyses of juvenile fishes need to be carefully conducted, considering the respective advantages and disadvantages of DNA-based and morphological techniques.     

Deoxyribonucleic Acid Homology Among Strains of Soybean-Nodulating Bacteria

Abstract

Characteristics of Brown Forest soils developed under different bio-climatic conditions from low to high eleyations in the Kinki District were studied with special reference to their pedogenetic processes. The Brown Forest soils at high elevations were characterized by a lower bulk density, higher capacities to adsorb organic matter, phosphate, and moisture, which were correlated with the ratio of the amorphous content to the clay content (the value of the ratio of (Feo + Alo)/clay), as compared to those at low elevations. Considering the fact that the value of the (Feo + Alo)/clay ratio was not correlated with the volcanic glass index, the formation of an amorphous fraction at high elevations was considered to proceed according to the following mechanism.

Low temperature at high elevations (above 700 m) may retard the crystallization of oxide minerals. The amorphous oxides with variable positive charge thus formed may adsorb organic matter, confering a darker color and high moisture and high phosphate retention capacities to the subsoil. Adsorbed organic matter stabilizes these amorphous oxides, thus enhancing the amorphous properties and inhibiting crystallization. A1 translocation due to the weak podzolization may contribute to the increase of the content of amorphous materials.

Characterization of the B horizons in forest soils in Japan, in terms of the values of the ratios of (Feo + Alo)/clay, (Fed-Feo)/Fet, and Alo/Ald, (CEC -ECEQ/CEC and STPT-ZPC, suggested that forest soils might be classified into four groups.     

Genetic relationships between the dominant genotypes of Phytophthora infestans in Hokkaido, Japan

The genetic characteristics of the dominant genotypes of Phytophthora infestans in Japan (US-1, JP-1, Japanese A1-A, A1-B) were compared. Differences were evident in the peptidase genotype, amplified fragment length polymorphism, and RG57 DNA fingerprints. Almost all of the fingerprint bands for the Japanese genotype A1-B were also present in JP-1 and Japanese A1-A, and few bands were unique to Japanese A1-B. These results suggest that the Japanese A1-B genotype was generated from sexual reproduction involving Japanese A1-A and JP-1 or related genotypes.     

Sexual mating preferences in vitro and in planta among Japanese isolates of Phytophthora infestans

The sexual preferences of Japanese isolates of Phytophthora infestans were determined by mating on agar, in broth, or in plants. The influence of their sexual preference was confirmed in the host tissues. Three wild-type isolates and a -glucuronidase (GUS) transformant were co-cultured to identify the origin of antheridia and oogonia. Japanese A1 isolate had a unique sexual preference compared with foreign isolates. It produced self-fertile oospores with about 40% of total gametangia but tended to form antheridia on V-8 agar medium. In addition, oospores were formed in plants, but their sexual preference could not be reflected in vitro.     

Molecular detection of protists associated with the Manila clam Ruditapes philippinarum inhabiting the Wajiro tidal flat in Hakata Bay,Fukuoka Prefecture

Fisheries Science - Tidal flats are ecologically important as they support a large community of animals (e.g., crabs, mollusks, and polychaetes) and restore water quality. However, information...     

Effects of oxidative stress caused by tert-butylhydroquinone on cytotoxicity in MDCK Cells

Antioxidant and oxidative stress effects of prooxidants are generally dose-dependent, and these effects depend on the prooxidant species and cell type. However, the cellular response to oxidant challenge is a complicated interplay of events involving cellular expression of phase II detoxification enzymes and cellular metal metabolism. This study demonstrates the effect of tert-butylhydroquinone (t-BHQ)-induced oxidative stress on MDCK cells. Cell toxicity tests were carried out using the crystal violet (CV) assay with the following prooxidants: t-BHQ, diethyl maleate (DEM), hydrogen peroxide (H(2)O(2)), diquat (DQ) and β-naphthoflavone (β-NF). Except for β-NF, these prooxidants showed dose-dependent cytotoxicity besides the most potent t-BHQ cytotoxicity. Only t-BHQ and DEM caused significant time-dependent expression of ferritin protein as an antioxidant, which segregates Fe(2+), causing the Fenton reaction. t-BHQ and DEM increased formation of lipid peroxidation, but DQ showed a tendency to decrease lipid peroxidation levels. In XTT assay, even when substantial cell death was observed in the CV assay, t-BHQ appeared to increase cell viability by enhancing XTT reduction, likely through the production of NADPH. Although curcumin, which induces cytoprotective phase II enzymes and chelates metal irons, decreased cell viability, it inhibited t-BHQ cytotoxicity. These results indicate that t-BHQ exhibits strong cytotoxicity against MDCK cells, an effect mitigated by curcumin, and that t-BHQ-induced oxidative stress activates the pentose phosphate pathway.     

Occurrence of orally administered mulberry 1-deoxynojirimycin in rat plasma

1-deoxynojirimycin (DNJ), a potent glucosidase inhibitor, is a characteristic constituent of the mulberry leaf. Dietary mulberry DNJ may be beneficial for the suppression of abnormally high blood glucose levels, thereby preventing diabetes mellitus. Although there is considerable interest in the effects of mulberry DNJ, the intestinal absorption and pharmacokinetic profile of orally administered mulberry DNJ have never been characterized. In this study, we developed a method for determining the level of plasma DNJ by hydrophilic interaction chromatography coupled to a mass spectrometric detector (HILIC-MS) to investigate the absorption and metabolism of orally administered mulberry DNJ in rats. DNJ was separated from plasma extract on a TSK gel Amide-80 column, a representative column for HILIC. At postcolumn, DNJ was concurrently detected and identified by MS. The plasma DNJ concentration in fasted rats was below the detection limit [<1 microg (6 nmol)/mL]; however, the concentration reached a maximum [15 microg (92 nmol)/mL] 30 min after the administration of mulberry DNJ (110 mg/kg of body weight), and the DNJ concentration decreased rapidly thereafter. When the rats received different amounts of mulberry DNJ (1.1, 11, and 110 mg/kg of body weight), dose-dependent incorporation of DNJ into the plasma was confirmed. We did not detect any DNJ metabolites in the plasma. These findings indicate that orally administered mulberry DNJ is absorbed as an intact form from the alimentary tract and then is quickly excreted from the body. The developed HILIC-MS method could be applied in determining levels of DNJ in urine and tissues, and therefore, the method would be a powerful tool for studying the metabolic fate of mulberry DNJ as well as its bioavailability.     

Determination of 1-deoxynojirimycin in mulberry leaves using hydrophilic interaction chromatography with evaporative light scattering detection

A simple and rapid method for determining 1-deoxynojirimycin (DNJ), a potent glucosidase inihibitor present in mulberry leaves (Morus alba and Morus bombysis), by high-performance liquid chromatography coupled to an evaporative light scattering detector (ELSD) has been developed. DNJ was separated from an extract of mulberry leaves on a TSKgel Amide-80 column, which is a representative column for hydrophilic interaction chromatography. During postcolumn detection, DNJ was detected by ELSD and concurrently identified by mass spectrometry. The detection limit was 100 ng. This method is sufficiently sensitive for determining DNJ in mulberry leaves and other related products.