Expression and purification of canine granulocyte colony-stimulating factor (cG-CSF)

Canine granulocyte colony-stimulating factor (cG-CSF) with modification of cysteine at position 17 to serine was expressed in Brevibacillus choshinensis HPD31. cG-CSF secreted into the culture medium was purified by ammonium sulfate precipitation and consecutive column chromatography, using butyl sepharose and DEAE sepharose. Biological activity of the recombinant cG-CSF was 8.0 × 10⁶ U/mg protein, as determined by its stimulatory effect on NFS-60 cell proliferation. Purified cG-CSF was subcutaneously administered once a day for two successive days to dogs (1, 5, 25, or 125 μg). Neutrophil count increased the following day in all dogs except those administered the lowest dose (1 μg). No severe side effects were observed in dogs after administration of cG-CSF.     

Suppression of male reproductive activity by estradiol in goldfish: appearance of infertile males that perform active spawning behavior

Fisheries Science - Estrogenic endocrine-disrupting chemicals are an increasing social concern, as they can reduce the reproductive activity of wildlife. The present study aimed to clarify the...     

Spatiotemporal distribution of Flavobacterium psychrophilum and ayu Plecoglossus altivelis in rivers revealed by environmental DNA analysis

Outbreaks of bacterial cold-water disease (BCWD), caused by Flavobacterium psychrophilum, are widespread in Japan, especially among ayu Plecoglossus altivelis. There are few investigations of F. psychrophilum in river water, and its seasonal distribution has not been clarified. We aimed to identify the spatiotemporal dynamics of F. psychrophilum and ayu to provide information that is useful for establishing a countermeasure for BCWD. Quantitative analysis of environmental DNA (eDNA) was used to clarify the year-round dynamics of ayu and F. psychrophilum. We sampled river water from the Nagara and Ibi rivers in Japan, and conducted monthly water sampling and eDNA quantification. Changes in the eDNA concentration of ayu were consistent with the known life histories of the fish. There was a strong negative correlation between the eDNA concentration of F. psychrophilum and water temperature, suggesting a strong dependence of F. psychrophilum dynamics in the river on water temperature. Furthermore, relatively high eDNA concentrations were recorded for both organisms in early summer and fall, suggesting that ayu is infected with F. psychrophilum during these seasons when experiencing up- and downmigration, respectively.

     

Impact of water percolation on nutrient leaching from an irrigated paddy field in Japan

Abstract. We examined the effect on soil nutrient status and sustainability of water percolation through an irrigated paddy field in Japan, to the depth of drainage (40 cm). The difference between amounts of nutrients leached by percolation and those supplied by irrigation indicated that 25–130 kg ha⁻¹ Ca, 8–24 kg ha⁻¹ Mg, from −1 to 9 kg ha⁻¹ K, and 8–17 kg ha⁻¹ Fe, respectively, were lost each year from the 0–40 cm soil layer during rice cultivation, when the supply from fertilization and rainfall and the loss in grain harvest were not accounted for. When the supply of K from rainfall and the loss in grain harvest were taken into account, a total K loss of about 10 kg ha⁻¹ was estimated. The electrical neutrality of inorganic ions in the percolating water was always maintained. From these results we estimate that the amounts of exchangeable Ca and Mg in the soil to a depth of 40 cm would decrease by 50% within 50–260 and 30–100 years, respectively, if similar management were continued without fertilization. The total amount of carbon dioxide (ΣCO₂) leached in percolating water during the period of rice cultivation was 120–325 kg C ha⁻¹, which corresponded to 0.47–0.94% of the soil organic carbon to 40 cm depth.     

Immunohistochemical and ultrastructural study of ito cells (fat-storing cells) in response to extrahepatic bile duct ligation in broiler chickens.

The Ito cell (fat-storing cell) lies in perisinusoidal space of liver and has a variety of functions. We investigated the immunohistochemistry and ultrastructure of Ito cells in normal and cholestatic livers of broiler chickens. Immunohistochemistry demonstrated that Ito cells expressed HHF35 muscle actin, vimentin, desmin, glial fibrillary acidic protein (GFAP), neuron-specific enolase (NSE), chromogranin A and cytokeratins in normal livers. These cells were diffusely scattered throughout the lobules. Livers treated with extrahepatic bile duct ligation (BDL) showed cholestasis, fibrosis, proliferation of biliary ductules and Ito cells. The Ito cells were frequently found in fibrotic areas and were larger in size with more extensive immunoreactivity than those of normal livers. Ultrastructural study demonstrated that Ito cells were closely associated with the production of collagen fibers in BDL livers. These findings suggest that Ito cells actively react against hepatocytic injuries and play a major role in the hepatic fibrogenesis of cholestatic livers of chickens.     

Accumulation of radioactive cesium in sorghum (Sorghum bicolor (L.) Moench) accessions cultivated in Fukushima in 2011 and 2012

ABSTRACT

The accident at Fukushima 1 Nuclear Power Plant in 2011 caused contamination by radioactive cesium (¹³⁴Cs and ¹³⁷Cs) in surrounding areas. After this accident, concerns about Cs contamination, including food safety, have limited industrial activities and reconstruction in Fukushima. Sorghum (Sorghum bicolor (L.) Moench) is an annual C₄ crop that can be used as biofuel feedstock due to its high biomass. Use of Cs-contaminated fields to produce biofuel feedstock would be more acceptable than use for food or feed crops due to the lower risk of human internal exposure to radioactive Cs. In addition, high-biomass sorghum might be suitable for removal of Cs from fields (phytoremediation). For both applications, it is important to use accessions showing the appropriate level of accumulation of radioactive Cs (low for biofuel feedstock, high for phytoremediation). Here, we examined the accumulation of radioactive Cs in the aerial parts of 56 sorghum accessions grown in Fukushima. Accessions were cultivated in a low-level-contaminated field in 2011 and in a highly contaminated field in a planned evacuation zone in 2012. After cultivation, activity concentrations from ¹³⁴Cs and ¹³⁷Cs were measured in the aerial plant parts. In 2011, the activity concentrations of ¹³⁴Cs and ¹³⁷Cs were 58.2–350 and 58.6–450 Bq kg^?1 dry weight, respectively. In 2012, the concentrations of ¹³⁴Cs and ¹³⁷Cs were 54.2–1320 and 57.1–1960 Bq kg^?1 dry weight, respectively. Relative to the median values of the accessions grown each year, 3 showed lower activity concentrations and 2 showed higher activity concentrations of radioactive Cs under both cultivation conditions. In contrast to a previous report, there was no significant correlation between biomass and Cs activity concentration. Because both biomass and Cs concentration are important in classifying accessions for use in phytoremediation, we also calculated the Cs accumulation index (single-plant biomass × Cs activity concentration) for each accession. The accession AKLMOI WHITE showed the highest values in both years, suggesting that this accession has the high per-plant accumulation capacity for radioactive Cs. Our data collected from actual contaminated fields is irreplaceable for choosing sorghum accessions for cultivation in Cs-polluted sites such as Fukushima.     

Seasonal changes of hormones and muscle enzymes in adult lacustrine masu (Oncorhynchus masou) and sockeye salmon (O. nerka)

The upstream migration of adult anadromous fishes is characterized by physiological changes in responses to reproductive and energetic challenges. This study analyzed the physiological responses of lake-resident anadromous masu salmon (Oncorhynchus masou) and sockeye salmon (O. nerka) to migration in order to determine if these fish might serve as a suitable model for ocean-running populations and to differentiate between physiological responses to reproduction and to exercise-linked aspects of migration. Reproductive (estradiol, testosterone, 11-ketotestosterone, 17α,20β-dihydroxy-4-pregnen-3-one) and metabolically-linked (thyroxine, triiodothyronine) hormones showed similar patterns to ocean-running anadromous populations. White muscle pyruvate kinase, lactate dehydrogenase and malate dehydrogenase decreased with the onset of spawning season while white muscle citrate synthase, β-hydroxyacetyl CoA dehydrogenase, phosphofructokinase and glutamate oxaloacetate transaminase did not, suggesting that the former group of enzymes are responding to reproductive or food intake signals while the second group, which typically change during anadromous migration, may be responding to exercise-linked aspects of migration. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effects of copper on survival and growth of larval false clown anemonefish <Emphasis Type="Italic">Amphiprion ocellaris</Emphasis>

ABSTRACT:   Larval false clown anemonefish of about 12 h after hatching were fed newly-hatched brine shrimp nauplii in natural (control) and copper-added seawater (40–640 µg–Cu/L) at 26.5 ± 0.5°C for 14 days. Survival rate of the fish increased with increasing copper concentrations up to 160 µg–Cu/L. The rates at 80 and 160 µg–Cu/L were 65 and 80%, respectively, and were significantly higher than that of the control (30%). A positive effect of copper addition on the survival rate was also observed in the other rearing experiment with the fish from seven different spawning chances. Growth rate of the survival fish was not affected by copper concentrations. The copper concentration of the survived fish increased with increasing copper concentrations in the rearing water.     

Effect of Feeding Frequency, Water Temperature, and Stocking Density on the Growth of Tiger Puffer, Takifugu rubripes

Effects of daily feeding frequency, water temperature, and stocking density on the growth of tiger puffer, Takifugu rubripes, fry were examined to develop effective techniques to produce tiger puffer in a closed recirculation system. Fish of 4, 14, and 180 g in initial body weight were fed commercial pellet diets once to five times a day to apparent satiation each by hand for 8 or 12 wk at 20 C. Daily feeding frequency did not affect the growth of 14‐ and 180‐g‐size fish. However, the daily feed consumption and weight gain of the 4‐g‐size fish fed three and five times daily were significantly higher than those of fish fed once daily (P < 0.05). Fish of 4 and 50 g in initial body weight were reared with the pellet diet at 15–30 C for 8 wk. The weight gain of fish increased with increasing water temperature up to 25 C and decreased drastically at 30 C for both sizes. Similar trends were observed for feed efficiency, although 4‐g fish had highest efficiency at 20 C. Effects of stocking density on growth were examined with fish of 8, 13, and 100 g in initial body weight. Fish were reared with the pellet diet for 8 or 16 wk at 20 C. Fish were placed in floating net cages in the culture tank, and the stocking density was determined based on the total weight of fish and volume of the net cage. Fish of 8 g in body weight grew up to 35–36 g during the 8‐wk rearing period independent of the stocking density of 8, 15, and 31 kg/m³ at the end of rearing. Final biomass per cage reached 32, 60, and 115 kg/m³ for 13‐g‐size fish, and 10, 18, and 35 kg/m³ for 100‐g‐size fish, and the growth of the fish tended to decrease with increasing stocking density for both sizes.     

Architecture of the photosynthetic oxygen-evolving center

Photosynthesis uses light energy to drive the oxidation of water at an oxygen-evolving catalytic site within photosystem II (PSII). We report the structure of PSII of the cyanobacterium Thermosynechococcus elongatus at 3.5 angstrom resolution. We have assigned most of the amino acid residues of this 650-kilodalton dimeric multisubunit complex and refined the structure to reveal its molecular architecture. Consequently, we are able to describe details of the binding sites for cofactors and propose a structure of the oxygen-evolving center (OEC). The data strongly suggest that the OEC contains a cubane-like Mn3CaO4 cluster linked to a fourth Mn by a mono-micro-oxo bridge. The details of the surrounding coordination sphere of the metal cluster and the implications for a possible oxygen-evolving mechanism are discussed.