Activities of Enzymes Related to the Malate–Aspartate Shuttle in the Blood Cells of Thoroughbred Horses Undergoing Training Exercise

The activities of the enzymes related to the malate–aspartate shuttle, which convert cytosolic NADH into mitochondrial NADH, were measured in red and white blood cells from thoroughbred horses undergoing continuous training (race horses) and compared with those in blood cells from riding horses. The activities of malate dehydrogenase (MDH), a rate-limiting enzyme for the malate-aspartate shuttle, were significantly elevated in the white blood cells (WBC) from race horses compared with those from riding horses. There were no significant differences in the activities of the enzymes in the red blood cells between race horses and riding horses. This increase in the MDH activity in their WBC is considered to reflect the increased metabolic activity in the race horses resulting from the training.     

Mechanisms responsible for reduced cardiotoxicity of mitoxantrone compared to doxorubicin examined in isolated guinea-pig heart preparations

We reported previously that doxorubicin, an anticancer agent that has an anthracycline structure, alters Ca2+ releasing and uptake mechanisms in the sarcoplasmic reticulum of myocardial cells. These effects of doxorubicin are apparently related to its cardiotoxicity. Mitoxantrone is a similar anticancer agent with an anthracenedion structure that has been shown to be significantly less cardiotoxic. In the present study, the effects of mitoxantrone on the functions of the sarcoplasmic reticulum were examined in isolated muscle preparations obtained from the guinea-pig heart. In electrically-stimulated left atrial muscle preparations, incubation in vitro for 4 hr with 30 or 100 microM mitoxantrone significantly prolonged the time to the peak of twitch tension, markedly increased the developed tension observed at lower stimulation frequencies, thereby attenuating the slope of positive force-frequency relationships, and increased the postrest contraction observed after a 60-sec quiescent period. In myocytes isolated from ventricular muscles, 30 microM mitoxantrone increased the peak and the size of intracellular Ca2+ concentrations ([Ca2+] i), and prolonged the time to peak [Ca2+]i. In skinned muscle fiber preparations obtained from the left ventricular muscle, 30 muM mitoxantrone significantly increased the caffeine-induced contraction without affecting the Ca2+ sensitivity of contractile proteins. These results suggest that mitoxantrone enhances Ca2+ release from the sarcoplasmic reticulum in isolated atrial muscle preparations obtained from the guinea-pig heart. Apparent enhancement of the sarcoplasmic reticulum functions, in contrast to anthracyclines that has been shown to suppress these functions, seems to explain the relative lack of marked cardiotoxicity of mitoxantrone.     

Seasonal fluctuation of groundwater in an evergreen forest, central Cambodia: experiments and two-dimensional numerical analysis

This study of a water cycle was conducted in an evergreen forest located in the Mekong River Basin in central Cambodia. At the observation site, we measured the dynamics of the spatial distribution of groundwater levels. The groundwater movement was analyzed two-dimensionally using boundary conditions and parameters that had been observed in the field. The climate in the research area is dominated by two seasons, which occur annually: a rainy and a dry season. The groundwater levels are generally high during the rainy season and low during the dry season. Groundwater levels were measured along a stream, which flowed through the study site. The streambed was visible at the head of the stream in January. At the next downriver well point, the streambed appeared in March. Finally, it became visible at all well points in April, meaning that surface runoff had disappeared temporarily and instead flowed underground during the ensuing dry period. Groundwater levels of the studied lateral flow perpendicular to the stream that seeped and infiltrated into the stream were 1.2–2.5 m deep (in April), which was the lowest level recorded for the year. During that period, the depth of the groundwater of the studied lateral flow fell by as much as 56 mm per month. In addition, the lateral flow groundwater infiltrated into groundwater of the stream during that period. The groundwater level fluctuation was estimated based on a two-dimensional analysis of lateral flow perpendicular to the stream using a numerical simulation model with soil physical parameters and observed boundary conditions. The observations of ground water fluctuations were well reproduced. Deep seepage of groundwater was estimated using a uniform boundary condition that allowed efflux through the bottom, estimated as being approximately 30 mm per year. The simulated deep seepage rate was considered plausible considering other hydrological components such as soil water storage fluctuation.     

Preparation of nori Pyropia yezoensis enriched with free amino acids by aging the culture with nori koji

Koji is commonly used in manufacturing Japanese fermented products and promotes enzymatic degradation. In recent years, a seaweed koji has been developed by culturing Aspergillus oryzae on nori Pyropia yezoensis. In the present study, we prepared nori koji by culturing A. oryzae on high- and low-quality noris and then mixing this preparation with additional dried nori. Suitable fermentation conditions for increasing the taste-active components of the nori mixed with nori koji were investigated. The mixture of nori and koji prepared from high-quality nori, with added water, cultured for 120 days at 10 °C, provided the greatest increase (7 times) in free amino acids. The changes in taste after culturing the nori and koji mixture were evaluated using a taste-sensing system. The sourness score of the nori and koji mixture increased significantly, but the scores for other attributes such as bitterness and umami richness did not increase after culture. The present study has demonstrated a clear increase in the free amino acid content of nori and a modification in the taste score by aging the culture with nori koji. These results will encourage the development of ‘aged seaweed,’ a novel value-added product with nutritional and taste elements modified using seaweed koji.

     

Differentiated predation risk on hatchery-reared juvenile masu salmon by white-spotted charr with different body sizes

Predation after release is one of the major concerns of hatchery fish propagation. However, size-specific interaction between predator and prey on the survival of hatchery-released salmonid fish is largely unknown. To understand the size-selective predation risk, 24-h predation experiments were conducted on masu salmon Oncorhynchus masou in tanks. Four ranges of fork length (FL) were examined for masu salmon as a prey, in combination with three ranges of FL for white-spotted charr Salvelinus leucomaenis as a predator. The results show that not only predator and prey sizes, but also interaction between prey size and predator size, strongly affected the survival rate of masu salmon. Predation on masu salmon with the FL exceeding 40% of the FL of white-spotted charr was rare in the experiment. A logistic regression suggests that 37% relative FL of masu salmon to white-spotted charr results in the 50% survival of masu salmon. Our results suggest that adjusting relative size of hatchery fish to the size of local fish predators at the time of hatchery release will have a significant impact on the survival of hatchery fish in the wild. From this perspective, site-specific, adaptive management might be important to improve the effectiveness of hatchery fish propagation.     

Estimation of whole-stem respiration,incorporating vertical and seasonal variations in stem CO<Subscript>2</Subscript> efflux rate,of <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> trees

We examined vertical and seasonal variations in stem respiration rates in a 50-year-old plantation of Japanese cypress, Chamaecyparis obtusa (Sieb. et Zucc.) Endl., in central Japan, and discuss a practical and precise method to scale a point-measured stem CO₂ efflux rate up to whole-stem respiration. For five selected trees, stem CO₂ efflux rates were measured at breast height (1.3 m) and at five or six points above breast height (at approximately 2 m intervals) every 1 or 2 months over two consecutive years. Daily total stem respiration rate (surface area basis) was greater inside the crown than below the crown, especially during the growing season. By incorporating the vertical profile of the respiration rate, annual whole-stem respiration was estimated for each sample tree (R _y). We then compared this estimate (R _y) with another estimate of annual whole-stem respiration (R′_y) obtained using a conventional method; it is assumed that the area-based respiration rate at breast height is constant throughout the stem. The ratio of these two estimates (R′_y/R _y) was usually less than 1, indicating that the assumptions used to calculate R′_y underestimate annual whole-stem respiration. We found that R′_y/R _y was negatively correlated with the ratio of crown length to tree height (crown ratio). These results suggest that annual whole-stem respiration in this C. obtusa plantation is substantially affected by the relative proportion of within-crown stem with higher respiratory activity. Methodologically, our results imply that incorporating the crown ratio into the conventional method would improve the accuracy of annual whole-stem respiration estimates.     

Supplemental effect of <Emphasis Type="Italic">Porphyra</Emphasis> spheroplasts on the growth and feed utilization of black sea bream

ABSTRACT:   A 56-day feeding trial was conducted to evaluate the effects of Porphyra spheroplasts (PS) as a feed additive on growth, carcass composition and feed utilization of black sea bream. Four experimental diets with or without PS (0, 1, 3, 5% inclusion level) were formulated to determine the proper incorporation level for best growth and utilization using white fishmeal as the principal protein source. Three replicate groups of fish averaging 1.28 ± 0.05 g (mean ± SD) were maintained in flow-through thermo-controlled sea water (32–34 psu, 25°C) aquaria (100 L) under the laboratory conditions (L12 : D12) and fed three times a day at 09:00, 13:00 and 17:00 hours by hand at approximately 3% body weight on each diet. The rearing trial revealed that growth performance (in terms of weight gain, specific growth rate) are almost the same level in all the dietary groups, nevertheless the survival, nutrient utilizations and retentions (feed efficiency, protein efficiency ratio, protein retention rate, lipid retention rate) were significantly higher ( P  < 0.05) in a group fed on the diet containing 3% spheroplasts. Further, the fish fed on the diet containing PS had significantly higher ( P  < 0.05) crude lipid level in their dorsal muscles and viscera. The fish fed a PS-based diet showed higher level of eicosapentaenoic acid and docosahexaenoic acid (DHA) in viscera and DHA in dorsal muscles. The results obtained in this feeding trial suggest that the optimum dietary PS supplementation level up to 3% for black sea bream can be considered as improved consequence on growth performance, nutrient utilization or body composition were noticed.     

稀释对CO掺氢混合燃料燃烧与排放性能的影响

在一台转速固定的火花点火发动机上进行了CO2稀释对CO掺氢混合燃料的燃烧及排放性能影响的试验研究.结果表明,氢气较快的燃烧速度弥补了由于其热值较低造成的对平均有效压力的影响,使得在每个当量比条件下平均有效压力基本保持不变.同时,掺氢对提升热效率及燃烧稳定性、改善CO以及NOx排放都有显著作用.适当的CO2稀释对平均有效压力和热效率的影响均不明显,而NOx排放水平则显著下降.但是过高的稀释率会导致部分燃烧,造成平均有效压力、热效率及CO排放的恶化.     

Fish meal replacement by soy protein from soymilk in the diets of red sea bream (Pagrus major)

Six isoenergetic diets were formulated as follows: fish meal (FM) 700 g kg^–1 (control, C), FM 300 g kg^–1 + soy protein concentrate 300 g kg^–1 (SPC), FM 300 g kg^–1 + enzyme‐treated SPC 300 g kg^–1 (ESC), FM 170 g kg^–1 + soy protein isolate 300 g kg^–1 (SPI), FM 160 g kg^–1 + enzyme‐treated SPI 300 g kg^–1 (ESI) and FM 150 g kg^–1 + conglycinin 300 g kg^–1(CG). Forty fish (3.9 g) were randomly distributed into each of eighteen 300‐L tanks, fed twice daily until satiation for 8 weeks. The final body weight, specific growth rate and condition factor did not show significant differences among the fish fed with diets C, SPC, ESC and ESI (p > .05). The survival was significantly lower in fish fed with diets SPI and CG. Feed efficiency was significantly higher in fish fed with diets SPC and C than in fish fed with other diets (p < .05). There were no significant differences in nutrients retention efficiencies in fish fed with diets C, SPC, ESC and ESI. A significantly higher phosphorus retention efficiency in fish fed with soymilk protein diets resulted in lower phosphorus discharge to the environment (p < .05). These results suggest that the soymilk proteins can comfortably replace 570–770 g FM kg^–1 diet of red sea bream juvenile, which will ensure significant ecological benefits through reducing phosphorus load to the environment.     

Availability of culture filtrate protein-10 (CFP-10) secreted from Mycobacterium pseudoshottsii for mycobacteriosis diagnosis in ginbuna crucian carp Carrasius auratus langsdorfii

Mycobacteriosis in cultured fish is a challenge for the aquaculture industry worldwide. Treatment by chemical administration is difficult and no effective vaccine has been developed. Therefore, detection and isolation by early diagnosis are important for prevention of the spread of the disease. In mammals, interferon gamma release assays have been established for detection of tuberculosis; these tests are based on the delayed-type hypersensitivity (DTH) response against culture filtrate protein-10 (CFP-10) and the 6-kDa early secreted antigen target (ESAT-6) of Mycobacterium tuberculosis. On the other hand, little is known about the fish immune response against the ESAT-6 and CFP-10 proteins of mycobacteria, although these responses should find application in the diagnosis of mycobacteriosis in fish. In the present study, we identified ESAT-6 and CFP-10 from Mycobacterium pseudoshottsii and cloned the corresponding genes. Intraperitoneal injection of the corresponding DNA plasmid constructs in ginbuna crucian carp yielded increased expression of the fish interferon-γ1-1-encoding gene (IFN-γ1-1). In contrast, IFN-γ1-1 expression accompanied by DTH response was observed only in the CFP-10-DNA plasmid-injected fish. Furthermore, fish that had been prophylactically injected with CFP-10-DNA plasmid exhibited increased survival of M. pseudoshottsii infection. Taken together, these results suggested that CFP-10 may facilitate diagnosis of mycobacteriosis.