In vivo and in vitro effects of Rhizopus extract on body growth and steroid production in masu salmon, Oncorhynchus masou Brevoort

This study investigated the in vivo and in vitro effects of Rhizopus (filamentous fungus) extract (RU) in masu salmon Oncorhynchus masou Brevoort. Underyearling fish were fed with RU for 16 months. Monthly changes in body growth, gonadal maturation and serum levels of sex steroids were monitored. Gonads were also incubated at 0, 1, 10, 100 and 1000 μg RU mL⁻¹ Leibovitz's L‐15 medium for 18 h. The levels of steroids in serum and cultured medium were measured. It was determined that RU‐fed immature and mature males, when compared with control groups, showed significantly higher body growth during spring, summer and the spawning period. Similarly, immature RU‐fed females showed significantly higher fork length and body weight in autumn, spring and summer. Furthermore, RU‐fed males showed significantly higher levels of serum testosterone (T) and 11‐ketotestosterone (11‐KT) levels in the pre‐spawning season, and 17α, 20β‐dihydroxy‐4‐pregnen‐3‐one (DHP) in the spawning season. In vitro RU incubation of gonads showed a dose‐dependent and significant increase in T, 11‐KT, oestradiol‐17β and DHP release in the medium. It appears that the causes of enhanced body growth and increased steroid production herein observed in salmonids are the physiologically active substances that may be present in the mycelium of the fungus.     

Nutritional evaluation of rotifers in rearing tanks without water exchange during seed production of amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis>

We evaluated the nutrient contents of rotifers sampled from larval-rearing tanks (tank rotifers) without water exchange during the seed production of amberjack Seriola dumerili at three facilities (Kamiura, Kagoshima, and Miyazaki) and compared them with the nutrient contents of freshly enriched rotifers. Compared to the enriched rotifers, the lipid contents, especially neutral lipids and proportion of 22:6n-3, tended to decrease in the tank rotifers. These trends were clearer at Miyazaki where the tank rotifers were sampled before daily supplementation of microalgae (Nannochloropsis). Crude protein content of the tank rotifers did not decrease markedly although the proportion of lysine tended to decrease. Vitamin C and E contents of the tank rotifers decreased significantly only at Miyazaki. Calcium content of the tank rotifers increased at Kamiura and Miyazaki, and the increases in iron and manganese contents of the tank rotifers at Miyazaki and zinc content at Kagoshima were pronounced. These results suggest that the nutritional value of rotifers in larval-rearing tanks without water exchange can be maintained by appropriate supplementation of microalgae. The effect of certain minerals that became high in tank rotifers on subsequent larval development requires further investigation.     

Vertical soil–air CO2 dynamics at the Takayama deciduous broadleaved forest AsiaFlux site

At the Takayama deciduous broadleaved forest Asiaflux site in Japan, the ecosystem carbon dynamics have been studied for more than two decades. In 2005, we installed non-dispersive infrared CO₂ sensors in the soil below the site’s flux tower to systematically study vertical soil–air CO₂ dynamics and explain the behavior of soil surface CO₂ efflux. Soil–air CO₂ concentrations measured from June 2005 through May 2006 showed sinusoidal variation, with maxima in July and minima in winter, similar to the soil CO₂ effluxes measured simultaneously using open-flow chambers. Soil–air CO₂ concentrations increased with soil depth from 5 to 50 cm: from 2,000 to 8,000 ppm in the summer and from 2,000 to 3,000 ppm in the winter under snow. Summer soil–air CO₂ concentrations were positively correlated with soil moisture on daily and weekly scales, indicating that the Oi, Oe, and A horizons, where decomposition is accelerated by high-moisture conditions, contributed substantially to CO₂ emissions. This result is consistent with the short residence time (about 2 h) of CO₂ in the soil and larger emissions in shallow soil layers based on our diffusion model. We revealed for the first time that soil–air CO₂ concentrations in winter were correlated with both snow depth and wind speed. CO₂ transfer through the snow was hundreds of times the gas diffusion rates in the soil. Our estimate of the CO₂ efflux during the snow-cover season was larger than previous estimates at TKY, and confirmed the important contribution of the snow-cover season to the site’s carbon dynamics.     

Localization of cytokines in tendinocytes of the superficial digital flexor tendon in the horse

Although inflammatory activation of cytokines have been analyzed in various tissues, there have only been a few and as-yet-inconclusive studies on cytokines in equine tendons. In this study, the localizations of 4 cytokines (IL-1alpha, IL-1beta, TNFalpha and IFNgamma) in tendinocytes of the equine superficial digital flexor tendon (SDFT) were analyzed by the use of an immunohistochemical method. In inflamed tendons positive staining for all 4 cytokines antibodies were detected in endotedinieum cells and vascular epithelial cells. In contrast, negative or trace immunoreactions were obtained in many tendinocytes in the normal tendon. The variation in cellular immune responses depending on the kind of cytokine may reflect the physiological/pathological condition of the SDFT.     

Alteration in the apoptosis process of rat esophageal epithelium with hyperproliferation of indigenous bacteria under a physiological condition

The apoptosis process in rat esophageal epithelium was investigated using enzyme-immunohistochemistry and transmission electron microscopy. As a result, Fas and Fas-L were expressed in the epithelial cell membrane and cytoplasm from the stratum spinosum (SS) to the stratum granulosum (SG). No TNF-R1 show immunopositivity in the cell membranes. TNF-α and caspase-8 were not observed in any layer. Caspase-10, cleaved caspase-3, XIAP and DNase-1 were found in the epithelial cytoplasm from the SS to the SG, whereas Bid, Apaf-1 and cleaved caspase-9 were detected only in the SG. Cytochrome c was observed as cytoplasmic granular positivity from the stratum basale (SB) and altered into homogeneous immunopositivity in the SG. Bcl-2 and Bcl-X immunopositivity was detected in cytoplasm from the SB to the SG. Immunoreactions of Bak in the cytoplasm and Bax beneath the cell membrane were observed from the upper portion of the SS with increasing intensity toward the SG. In the sites with the hyperproliferation of indigenous bacteria, TNF-R1, TNF-α and caspase-8 were detected in the SG and the immunopositive intensities of Bid, Apaf-1 and cleaved caspase-9 were altered to be strong. Prominently swollen cells and decreased mitochondria were ultrastructurally confirmed in the uppermost layers of stratum corneum. These findings suggest that the Fas-Fas-L-interaction initially induces apoptosis through a mitochondria-independent pathway and secondarily through a mitochondria-dependent pathway, leading to eventual epithelial cell death in the rat esophageal epithelium. The bacterial stimuli probably enhance the mitochondria-dependent pathway through the TNF-R1-TNF-α interaction.