Characterization of the goldfish fecal microflora by the fluorescent in situ hybridization method

ABSTRACT:   Bacterial populations in goldfish feces were characterized by the fluorescent in situ hybridization (FISH) method. A total of nine different group-specific rRNA-targeted oligonucleotide probes were used. Approximately half of the microbial cells (57.8 ± 16.7%) were detected with a probe EUB338 and found to be bacteria. The microbial cells in 33–35 of the 35 samples from five specimens strongly hybridized with probes ALF1b, BET42a and GAM42a, suggesting that goldfish intestinal bacteria are mainly composed of α, β and γ-subclasses of Proteobacteria. The fact that a probe AER66 reacted with 25.6 ± 14.2% of the total microbial cells in all 35 samples, demonstrated that genus Aeromonas was the dominant species in the goldfish intestines. Genus Bacteroides including Bacteroides type A detected with a probe BAC303 was observed in 15 of 35 samples while other taxonomic groups determined with HGC69a, CF39a and P72 were detected in 6–11 of 35 samples. These results strongly suggest that Bacteroides shows the greatest daily fluctuation and interindividual variation in the intestines of goldfish. Moreover, the FISH method was proven to be useful for rapid enumeration of taxonomic groups of fish intestinal bacteria.     

Isolation, identification and characterization of disomic and translocated barley chromosome addition lines of common wheat

Two disomic barley chromosome addition lines and five translocated chromosome addition lines of common wheat cultivar Shinchunaga were isolated. They were derived from a hybrid plant between Shinchunaga and cultivated barley Nyugoruden (New Golden) by backcrossing with wheat and self pollination. Barley chromosomes added to chromosome arms involved in the translocated chromosomes were identified by C-banding method and by crossing these lines with Chinese Spring/Betzes addition lines. Two disomic addition lines were identified to have chromosome 6 and 7 of barley, respectively. Two of the five translocated chromosome addition lines were clarified to have same chromosome constitution, 42 wheat chromosomes and a pair of translocated chromosomes constituted with a long arm of chromosome 5B of wheat and a short arm of chromosome 7 of barley. The other three lines could not be identified due to chromosome rearrangement. Performances of these seven lines on agronomic characters were examined. Addition of barley chromosome 7 induced early heading, and chromosome 6 showed lated heading. Almost all of the lines except that of chromosome 6 showed short culm length and all showed reduced number of tillers, spikelets and grains per ear, and low seed fertility. These lines would be useful for genetic analyses in wheat and barley and for induction of useful genes of barley into wheat. This revised version was published online in July 2006 with corrections to the Cover Date.     

A specific and potent inhibitor of brassinosteroid biosynthesis possessing a dioxolane ring

Screening for brassinosteroid biosynthesis inhibitors was performed to find azole derivatives that induced dwarfism, to resemble brassinosteroid-deficient mutants in Arabidopsis, and which could be rescued by brassinosteroid. Through this screening experiment, propiconazole fungicide was selected as a likely inhibitor of brassinosteroid biosynthesis and, thus, propiconazole derivatives with optimized activity and selectivity were synthesized. The biological activity of these compounds was evaluated by examining cress stem elongation. Among the compounds tested, 2RS,4RS-1-[2-(4-trifluoromethylphenyl)-4-n-propyl-1,3-dioxolan-2-ylmethyl]-1H-1,2,4-triazole (12) showed the most potent capability to retard cress stem elongation in the light. The compound-induced hypocotyl dwarfism was restored by the coapplication of 10 nM brassinolide but not by 1 microM gibberellin. These results suggest that 12 should affect brassinosteroid biosynthesis. The potency and specificity of 12 were greater than those of brassinazole, a previously reported brassinosteroid biosynthesis inhibitor.     

Preparation of phytate-removed deamidated soybean globulins by ion exchangers and characterization of their calcium-binding ability.

Phytate-removed deamidated soybean globulins were prepared using ion-exchange resins to provide them with a functional property to enhance calcium absorption in the body. The phosphorus level was reduced from 45 to 20 micromol/g using 0.05 g/mL of AE-4, an anion-exchange resin with a (2-hydroxyethyl)dimethylammonium group, in 0.2% soybean globulin solution for 1 h at 4 degrees C, and 90-92% of the phosphorus in defatted soybeans could be removed. As for deamidation, CE-4, a cation-exchange resin of the carboxylate type, showed a much higher deamidation activity than CE-1 and CE-2, cation-exchange resins of the sulfonate type. No peptide bond hydrolysis was observed for any cation-exchange resin treated at 4 degrees C. There was no significant difference in the amount of acid amide deamidated at temperatures between 4 and 50 degrees C. The deamidation level was able to increase to 73% using 0.10 g/mL of CE-4 in a 0.2% soybean globulin solution for 6 h at 4 degrees C. The amount of calcium bound to the soybean globulins decreased with removal of the phytate but increased with deamidation.     

Precise, simple screening for resistance in potato varieties to common scab using paper pots

Resistance to common scab pathogen Streptomyces turgidiscabies of seven potato varieties was compared in the field with a newly developed paper pot method. Seedlings raised in soil in paper pots containing inocula at 1 × 10³ to 10⁷cfu/g soil were transplanted into a scab-free field and grown for 3 months. The disease severity of the seven varieties in the field trials differed in iteration and from year to year, even though their resistance levels were approximately similar at the expected levels. With the paper pot method, the seven varieties had different resistance levels, which were almost completely consistent with the results of the field trials, at more than 1 × 10⁵cfu/g soil. Significant differences in disease severity between resistant and susceptible varieties were observed (P = 0.05) for 2 years, and the resistance level of the varieties was elucidated.     

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.     

Environmental factors influencing sporocarp formation in <Emphasis Type="Italic">Typhula ishikariensis</Emphasis>

Environmental factors influencing sporocarp formation in Typhula ishikariensis were studied under controlled conditions. Sporocarp formation in T. ishikariensis was divided into two stages: stipe elongation from the sclerotium and fertile head development at the tip of the stipe. Factors required for each stage differed. At the stipe elongation stage, low temperature (10°/5°C; day/night) and high humidity were important, but light was not required. In contrast, at the fertile head stage, light and moderate day length (8h/day) were essential. Fertile heads developed at 46µEm^–2s^–1; and high intensity (137µEm^–2s^–1) did not suppress development. Moreover, adding unsterilized soil to the sea sand medium accelerated sporocarp formation. These findings imply that the sclerotium of T. ishikariensis recognizes several physical factors for sporocarp formation. Sporocarps of T. ishikariensis developed within 4 weeks after incubation under optimal conditions. The sporocarp produced basidiospores, and differential mating incompatibility was confirmed among monokaryons derived from basidiospores produced under artificial conditions. This method should be useful for obtaining monokaryons for genetic studies of T. ishikariensis.     

Fibrodysplasia ossificans progressiva-like condition in a cat

Fibrodysplasia ossificans progressiva (FOP)-like condition was diagnosed in a Japanese domestic cat with stiffness, marked atrophy of the muscles, and limited mobility of all joints in both the pelvic limbs. Etretinate, a retinoid, was used for medical management; however, no improvement in the clinical signs was observed. Inheritance of the disorder has not yet been demonstrated. Furthermore, the clinical signs and histopathological findings of feline FOP-like condition in the present case differed from those of the previously reported cases.     

Plasma adrenomedullin concentration in dogs with myxomatous mitral valvular disease

Adrenomedullin (AM), a peptide identified to have vasodilating and natriuretic effects, is involved in the regulation of the cardiovascular system. To evaluate plasma AM concentration in dogs with myxomatous mitral valvular disease (MMVD), and to investigate the associations between the concentrations of plasma AM and natriuretic peptides and the echocardiographic data, we evaluated plasma AM concentrations in 31 healthy control dogs and 57 dogs with MMVD. Plasma AM concentrations in dogs with MMVD were higher than that in the control subjects. The plasma AM concentration increased in conjunction with the severity of heart failure according to the International Small Animal Cardiac Health Council (ISACHC). The AM concentrations were 25.1 ± 5.0 fmol/ml (ISACHC class Ia), 29.9 ± 11.0 fmol/ml (ISACHC class Ib), 43.4 ± 19.8 fmol/ml (ISACHC class II) and 73.5 ± 21.7 fmol/ml (ISACHC class III) and 7.5 ± 5.1 fmol/ml (control group), respectively. The receiver operating characteristic curve indicated an area of 0.93 (95% CI, 0.8801-0.9889; <0.0001), a cutoff value of 30.5 fmol/ml, a sensitivity of 87.1%, and a specificity of 82.5% for the determination of congestive heart failure. Plasma AM concentrations correlated with atrial natriuretic peptide concentrations, LA/Ao ratio, and left ventricular diameter. In conclusion, AM may be a potential diagnostic marker for canine MMVD and possibly plays a pathophysiological role in collaboration with the other neurohumoral factors such as natriuretic peptides.