Optimum culture conditions of <Emphasis Type="Italic">Rhodomonas</Emphasis> sp. Hf-1 strain as a live food for aquatic animals

Suitable culture conditions for Rhodomonas sp. Hf-1 strain were examined for high productivity. Hf-1 strain was grown in an incubator for 7 days. The factorial experimental design investigated the following 19 variables: temperature (12, 16, 20, 24 and 28 °C), salinity (7, 14, 21, 28 and 35 psu), light intensity (20, 35, 50, 65 and 80 μmol m^?2 s^?1), light color (white, red, green and blue lights), and 3 factorial designs of photoperiod (24L:0D, 16L:8D and 12L:12D light:dark cycle). The cell density and specific growth rates (SGR) were analyzed. The best cell growth was observed under the following culture conditions: temperature of 24 °C, salinity of 21 psu, light intensity of 80 μmol m^?2 s^?1, and white light. In the photoperiod test, the highest cell density of 4.7?×?10⁶ cells ml^?1 was obtained at 24L:0D light:dark cycle, and the SGR was 0.57 μ day^?1 at this time. We found that the Hf-1 strain was able to be cultured in extremely wide culture conditions. These results are expected to serve as a baseline study for culturing the Hf-1 strain in the laboratory and for its use in aquaculture.     

Identification of a 4-Deoxy-l-erythro-5-hexoseulose Uronic Acid Reductase,FlRed, in an Alginolytic Bacterium Flavobacterium sp. Strain UMI-01

In alginate-assimilating bacteria, alginate is depolymerized to unsaturated monosaccharide by the actions of endolytic and exolytic alginate lyases (EC 4.2.2.3 and EC 4.2.2.11). The monosaccharide is non-enzymatically converted to 4-deoxy-l-erythro-5-hexoseulose uronic acid (DEH), then reduced to 2-keto-3-deoxy-d-gluconate (KDG) by a specific reductase, and metabolized through the Entner–Doudoroff pathway. Recently, the NADPH-dependent reductase A1-R that belongs to short-chain dehydrogenases/reductases (SDR) superfamily was identified as the DEH-reductase in Sphingomonas sp. A1. We have subsequently noticed that an SDR-like enzyme gene, flred, occurred in the genome of an alginolytic bacterium Flavobacterium sp. strain UMI-01. In the present study, we report on the deduced amino-acid sequence of flred and DEH-reducing activity of recombinant FlRed. The deduced amino-acid sequence of flred comprised 254 residues and showed 34% amino-acid identities to that of A1-R from Sphingomonas sp. A1 and 80%–88% to those of SDR-like enzymes from several alginolytic bacteria. Common sequence motifs of SDR-superfamily enzymes, e.g., the catalytic tetrad Asn-Lys-Tyr-Ser and the cofactor-binding sequence Thr-Gly-x-x-x-Gly-x-Gly in Rossmann fold, were completely conserved in FlRed. On the other hand, an Arg residue that determined the NADPH-specificity of Sphingomonas A1-R was replaced by Glu in FlRed. Thus, we investigated cofactor-preference of FlRed using a recombinant enzyme. As a result, the recombinant FlRed (recFlRed) was found to show high specificity to NADH. recFlRed exhibited practically no activity toward variety of aldehyde, ketone, keto ester, keto acid and aldose substrates except for DEH. On the basis of these results, we conclude that FlRed is the NADH-dependent DEH-specific SDR of Flavobacterium sp. strain UMI-01.     

Exposure to blue LED light before the onset of darkness under a long‐day photoperiod alters melatonin secretion,feeding behaviour and growth in female dairy calves

The effect of blue LED on melatonin secretion, feeding behaviour and growth was addressed in Holstein female dairy calves. In Exp.1, six animals (8 weeks old, 97 ± 4.1 kg BW) were exposed to yellow or blue LED for 2 hr before darkness over 7 days under a long‐day photoperiod (LDPP). In Exp. 2, six animals (8 weeks old, 88.5 ± 4.8 kg BW) were exposed to blue light from a white LED all daytime or a yellow LED for 2 hr before the darkness of LDPP (blue light cut) over 3 weeks. In Exp. 1, blue light mildly suppressed melatonin secretion during the 2‐hr treatment but did not affect the timing of the nightly melatonin rise. However, the rise in nighty melatonin levels was higher with yellow than blue LED. In Exp. 2, white LED completely suppressed melatonin secretion during the 2‐hr treatment, but plasma melatonin concentrations were similar during the darkness. Grass hay intake, rumination time, frequency of water intake and body weight gain were higher in animals exposed to the yellow rather than the white LED. Overall results indicate that exposure to blue light from white LEDs under an LDPP suppresses melatonin secretion and might negatively impact the development of female dairy calves.     

Microbial reduction mechanism of ferric iron in paddy soils (Part I)

In the previous paper¹⁾ the authors proved that the ferric iron reduction in submerged paddy soils is entirely or mostly effected by the activities of microorganisms in the soil. Prior to the study on the ferric iron reducing microorganisms isolated from the soil, the present paper reports the research for the microbial mechanism of iron reduction. working with the soil itself.     

The reduction mechanism of manganese in paddy soils

Hydroquinone method manganese (soluble in pH 7.0, 1 N-ammonium acetate solution containing 0.2 percent hydroquinone) and microbially active manganese (soluble in pH 7.0, 1 M-magnesium sulfate solution after flooding soils with or without Chinese milk vetch for 12 or 20 days respectively at 30°C) of 22 paddy soils were determined. The amounts of manganese reduced with sodium oxalate under acid conditions (oxalate method manganese (a) and (b), the former was determined under more rigorous conditions than the latter) were also determined and compared with hydroquinone method manganese and microbially active manganese.

Their levels of many soil samples representing soil groups were also determined to examine the dlfferences In amounts of active manganese among soil groups. The results obtained are as follows.

The relationship between microbially and chemically active manganese. 1) The amounts of microbially active manganese in soils were 48 to 68 mg Mn per 100 g oven-dried soil and these were increased by the addition of Chinese milk vetch. 2) The amounts of hydroquinone method manganese were less than microbially active manganese, and the amounts of oxalate method manganese (b) were larger than microbially active manganese. The amounts of oxalate method manganese (a) were the largest of all the types of manganese. 3) There were high correlations between the amounts of various types of active manganese described in 2). The levels of microbially and chemically active manganese. 1) The amounts of microbially active manganese lay between the amounts of chemically active manganese determined by the hydroqulnone method and by the oxalate method (b) In all soil samples representing soil groups. High correlations were found between these types of active manganese. 2) The hydroquinone method was considered to be unsuitable for quantitatively determining the amounts of chemicallY active manganese in soils of high organic matter content. 3) In both cases of microbially active manganese and chemically active manganese, tha widest range and the largest amount determined were both observed in strongly gley soila. The averages of theae types of active manganese were high in strongly gley soils, pea, and muck soils, and black soils. The differences among soil groups were smalle1 than the differences among soil samples, and little tendency was observed in the differences among soil groups.

From these findings described above it is suggested that the oxalate methoo (b) is more appropriate than the hydroqulnone method for determining chemically active manganese as an index of microbially active manganese.     

Effect of flutianil on the morphology and gene expression of powdery mildew

Flutianil, a fungicide effective only on powdery mildew, was previously reported to affect the host cell''s haustorial formation and nutrient absorption. Studies were conducted to investigate flutianil''s primary site of action on Blumeria graminis morphology using transmission electron microscope (TEM) observation and RNA sequencing (RAN-seq) techniques. TEM observation revealed that flutianil caused the extra-haustorial matrix and fungal cell wall to be obscured, without remarkable changes of other fungal organelles. RNA-seq analysis indicated that, unlike other powdery-mildew fungicides, flutianil did not significantly affect the constantly expressed genes for the survival of B. graminis. Genes whose expression is up- or downregulated by flutianil were found; these are the three sugar transporter genes and various effector genes, mainly expressed in haustoria. These findings indicate that the primary site of action of flutianil might be in the haustoria.     

Antiproliferative and antiangiogenic activities of smenospongine, a marine sponge sesquiterpene aminoquinone

We previously reported that smenospongine, a sesquiterpene aminoquinone isolated from the marine sponge Dactylospongia elegans, showed antiproliferative or cytotoxic activities on leukemia cells. In this study, we investigated the effect of smenospongine on solid tumors. Since angiogenesis is well known to be closely involved in growth and metastasis of solid tumors, the antiangiogenic effect of smenospongine was determined. We found that smenospongine inhibited proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVEC). Moreover, the inhibitory activity of smenospongine on growth of solid tumor cells was investigated. Smenospongine inhibited the growth of 39 human solid cancer cells in vitro, with a mean Log GI(50) value of -5.55. In conclusion, smenospongine exhibits antitumor activity on solid tumors via two mechanisms, an antiangiogenic effect on endothelial cells and direct inhibition of growth of tumor cells.     

Squamous cell carcinoma in a digit of the hind limb with systemic metastasis in a 17-year-old female koala

We encountered a case of cutaneous squamous cell carcinoma (SCC) in a 17-year-old female koala at a zoo. A fragile, papillary, elevated mass was found on the third digit of the right hind limb. SCC was identified histopathologically: squamous cell-like polygonal tumor cells showed a nest-like growth pattern with epidermal down growth, central keratinization and necrotic foci, and invaded dermal connective tissues. Metastatic lesions were observed in various organs, including the lung and axillary lymph node: in the lung, multiple metastatic foci similar to the primary lesion, and in the axillary lymph node, individual polygonal tumor cells infiltrated the sinusoids. Immunohistochemistry revealed that the tumor cells were positive for proliferating cell nuclear antigen, which exhibited 32–33% of labeling indices in the tumor cells. To our knowledge, this is the first report of a case of SCC in a digit of a koala.     

Anatomical characterization of decayed wood in standing light red meranti and identification of the fungi isolated from the decayed area

To further our understanding of wood decay in living light red meranti (Shorea smithiana) trees, microscopic characteristics of the cell and cell wall degradations of S. smithiana wood in the presence of the decay fungi, the identity of the causal fungi, and the decay potential and pattern by an isolated fungus were investigated. Cell wall degradations, including cell wall thinning, bore holes formation, rounded pit erosion, and eroded channel opening were clearly observed under light and scanning electron microscopy. In transverse view, many large voids resulting from a coalition of degraded wood tissue appeared in the decayed canker zone. All these observations suggest the well-known simultaneous decay pattern caused by white-rot fungi. By phylogenetic analysis based on the sequences of internal transcribed spacer region of ribosomal DNA, a basidiomycete fungus isolated from the decayed wood was identified as Schizophyllum commune. The degradation caused by this fungus on sound S. smithiana wood in an in situ laboratory decay test was classified as the early stage of simultaneous decay, and showed a similar pattern to that observed in the wood samples naturally decayed.