Sarcoplasmic Reticulum Ca2+-ATPase Activity and Glycogen Content in Various Fiber Types after Intensive Exercise in Thoroughbred Horses

To find a new parameter indicating muscle fitness in Thoroughbred horses, we examined time-dependent recovery of glycogen content and sarcoplasmic reticulum (SR) Ca²⁺-ATPase activity of skeletal muscle after intensive treadmill running. Two repeated 50-sec running sessions (13 m/sec) were performed on a flat treadmill (approximately 90%VO₂max). Muscle samples of the middle gluteal muscle were taken before exercise (pre) and 1 min, 20 min, 60 min, and 24 hr after exercise. Muscle fiber type composition was determined in the pre muscle samples by immunohistochemical staining with monoclonal antibody to myosin heavy chain. SR Ca²⁺-ATPase activity of the muscle and glycogen content of each muscle fiber type were determined with biochemical analysis and quantitative histochemical staining, respectively. As compared to the pre value, the glycogen content of each muscle fiber type was reduced by 15–27% at 1 min, 20 min, and 60 min after the exercise and recovered to the pre value at 24 hr after exercise test. These results indicate that 24 hr is enough time to recover glycogen content after short-term intensive exercise. The mean value of the SR Ca²⁺-ATPase activity showed a slight decrease (not significant) immediately after exercise, and complete recovery at 60 min after exercise. There were no significant relationship between the changes in glycogen content of each muscle fiber type and SR Ca²⁺-ATPase. Although further studies are needed, SR Ca²⁺-ATPase is not a useful parameter to detect muscle fitness, at least in Thoroughbred horses.     

Effect of feeding rotifers enriched with taurine on the growth and survival of larval amberjack Seriola dumerili

The effect of feeding rotifers enriched with taurine on the growth performance and survival of larval amberjack Seriola dumerili was investigated. Rotifers were enriched with a commercial taurine supplement at four levels (0, 200, 400, and 800 mg/l). The larvae were fed the enriched rotifers in triplicate from 3 days post-hatch for 7 days under static conditions. The average taurine contents of the rotifers were 1.5, 2.7, 4.2, and 7.2 mg/g dry matter, respectively. The growth of the fish fed rotifers enriched with the taurine supplement at 800 mg/l was significantly (P < 0.05) improved compared with that of the fish fed the rotifers without taurine enrichment. The survival rate improved proportionally up to a taurine supplement level of 400 mg/l, but no significant differences in survival were observed among treatments. The fraction of the larvae with inflated swim bladders did not vary significantly between treatments. Taurine content in the whole fish body increased with the taurine level in the rotifers. These results suggest that taurine enrichment of rotifers is an effective method of enhancing the growth of amberjack larvae.     

Partial characterization and ontogenetic development of pancreatic digestive enzymes in Japanese eel Anguilla japonica larvae

The pancreatic digestive enzymes, trypsin, chymotrypsin, lipase and amylase were partially characterized, and changes in their activities were examined during the initial ontogeny of Japanese eel Anguilla japonica larvae from 5 to 34 days post-hatching (dph). The pH optima of the eel larval enzymes were narrower than those other fish species; trypsin activity was highest at pH 9, chymotrypsin and amylase activities were highest at pH 7 and 8, and lipase activity was highest at pH 8 and 9. In an analysis of thermal profiles, the larval pancreatic enzymes had a high optimal temperature and high thermal stability, which are typical of fish from the tropics. At 12 and 13 dph, lipase activity and gene expression levels of trypsin (-a and -b), lipase and amylase decreased markedly, suggesting a marked change in larval metabolism at that time. These data could be useful in the development of artificial larval diets in Japanese eel.     

Effects of docosahexaenoic acid on growth,survival and swim bladder inflation of larval amberjack ( Seriola dumerili, Risso)

The effect of docosahexaenoic acid (DHA) on the growth performance, survival and swim bladder inflation of larval Seriola dumerili during the rotifer feeding period was investigated in two feeding experiments. Amberjack larvae at 3 day post hatching were fed rotifers enriched with (1) freshwater C hlorella (Chlo), (2) a mixture (2:1, v/v) of Chlo and DHA‐enriched C hlorella (DHA‐Chlo), (3) DHA‐Chlo and (4) DHA‐Chlo and commercial DHA emulsion, in triplicate for 7 days. The average DHA contents of the rotifers were 0.0, 0.4, 1.0 and 1.9 mg g^?1 DM respectively. The survival rate was improved by the enrichment of rotifers with DHA‐Chlo alone, and DHA‐Chlo and emulsion. Growth and swim bladder inflation of fish fed rotifers enriched with DHA‐Chlo were significantly (P < 0.05) improved, however, with increased levels of DHA further improvement was not found. DHA content in the larval whole body proportionally increased with the DHA level in the rotifers. These results suggest that DHA enrichment of rotifers is effective to improve the growth, survival rate and swim bladder inflation of amberjack larvae. The DHA requirement of amberjack larvae is estimated to be 1.5 mg g^?1 on a dry matter basis of rotifers.     

Involvement of NbNOA1 in NO production and defense responses in INF1-treated Nicotiana benthamiana

During defense responses, plant cells produce nitric oxide (NO), which may control many physiological processes. In a previous study, we reported that nitrate reductase (NR) is responsible in part for INF1 elicitor-induced NO production in Nicotiana benthamiana, but the possibility remains that other NO-generating system(s) contribute to NO production. In mammalian cells, NO production is catalyzed by NO synthase (NOS). However, NOS-like enzyme(s) have never been identified in plants, and only the gene for Arabidopsis thaliana nitric oxide-associated 1 (AtNOA1) has been identified as a putative regulator of NOS activity in plants. In this study, we cloned NbNOA1, a homolog of AtNOA1, from N. benthamiana and investigated its involvement in NO production induced by INF1. The NbNOA1 gene was silenced by a virus-induced gene-silencing (VIGS) technique. NbNOA1-silenced plants had yellowish leaves. Silencing NbNOA1 partially decreased INF1-induced NO production, while overexpressing NbNOA1 did not affect NO production. Silencing NbNOA1 suppressed INF1-induced PR1a gene expression and increased susceptibility to Colletotrichum lagenarium. These results suggest that NbNOA1 is involved in INF1-mediated NO production and is required for defense responses. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB303300.     

Growth performance and physiological condition of F1 amago salmon Oncorhynchus masou ishikawae juveniles obtained from broodstock with selective breeding for growth on a low fish-meal diet

A 10-week feeding trial was conducted to evaluate the growth and physiological condition of F1 amago salmon Oncorhynchus masou ishikawae juveniles obtained from broodstock that had been selected for growth on a low fish-meal (LFM) diet. F1 juveniles from broodstock showing superior or inferior growth on a LFM diet (LFM-S, LFM-I), and those from broodstock showing intermediate growth on a fish meal-based (FM) diet (FM-M), were fed a LFM diet and a FM diet. In fish fed the LFM diet, growth of LFM-S was significantly better than FM-M at 3 weeks; however, this superiority in growth disappeared at the end of the 10-week trial. Growth of LFM-I was significantly lower than LFM-S throughout the feeding trial. Growth of fish fed the FM diet was not significantly different among groups, and higher than fish fed the LFM diet. Improvements in the blood hemoglobin concentration and morphological conditions of the distal intestine and liver were observed in LFM-S fed the LFM diet. Although the effect of selective breeding was limited in F1 amago salmon, the present results suggest that continuous selection for growth with a LFM diet has potential as a method to adapt to the limited supply of fish meal.     

Characterization and ontogenetic development of digestive enzymes in Pacific bluefin tuna Thunnus orientalis larvae

The major digestive enzymes in Pacific bluefin tuna Thunnus orientalis larvae were characterized, and the physiological characteristics of the enzymes during early ontogeny were clarified using biochemical and molecular approaches. The maximum activity of trypsin (Try), chymotrypsin (Ct) and amylase (Amy) was observed at pH 6–11, 8–11 and 6–9, respectively. Maximum activity of Try, Ct and Amy occurred at 50 °C, that of lipase (Lip) was at 60 °C and that of pepsin (Pep) was at 40–50 °C. These pH and thermal profiles were similar to those for other fish species but differed from those previously reported for adult bluefin tuna. Enzyme activity for all enzymes assayed was found to decrease at high temperatures (Try, Ct, Amy and Pep: 50 °C; Lip: 40 °C), which is similar to findings for other fish species with one marked exception—increased Try activity was observed at 40 °C. Lip activity appeared to be dependent on bile salts under our assay conditions, resulting in a significant increase in activity in the presence of bile salts. Ontogenetic changes in pancreatic digestive enzymes showed similar gene expression patterns to those of other fish species, whereas marked temporal increases in enzyme activities were observed at 10–12 days post hatching (dph), coinciding with previously reported timing of the development of the pyloric caeca in bluefin tuna larvae. However, complete development of digestive function was indicated by the high pep gene expression from 19 dph, which contradicts the profile of Pep activity and previously reported development timing of the gastric gland. These findings contribute to the general knowledge of bluefin tuna larval digestive system development.     

Effect of high molecular weight carbohydrates on bud cell formation by <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in potato dextrose broth

Potato dextrose broth (PDB), one of the most popular culture media for fungi, can be made in the laboratory from potato extract and glucose (handmade PDB) and also bought as powder (commercial PDB). We compared growth of Fusarium oxysporum in handmade and commercial PDBs. For seven strains tested, bud cell formation and fungal biomass were higher in handmade PDB than in commercial PDB. Gel filtration analyses revealed that handmade PDB contained carbohydrates with molecular weights >20 MDa, while most carbohydrates in commercial PDB were <45 kDa. Freshly prepared, raw potato starch had a molecular weight >20 MDa and enhanced bud cell formation by five of seven strains when added to commercial PDB. The addition of glucose to commercial PDB promoted biomass production but not bud cell formation. Furthermore, in a semisynthetic medium containing raw potato starch, bud cell formation was enhanced for all strains tested. These results indicate that the high molecular carbohydrates present in raw potatoes enhanced bud cell formation by Fusarium oxysporum.     

Strength properties of laminated veneer lumber in compression perpendicular to its grain

Tests of compression perpendicular to the grain were carried out on laminated veneer lumber (LVL) and timber. The species tested were sugi, radiata pine, karamatsu, akamatsu, and dahurian larch; two sets of sugi specimens were tested, with the sugi LVL products being manufactured in different plants. The strength properties of the materials for different loading directions were compared for LVL and timber. At 5% compressive strain in the same materials, the average stress in the tangential direction of timber was larger than that in the radial direction for all species except for radiata pine, and the average stress in the edge-wise direction of LVL was larger than that in the flat-wise direction for all species except for radiata pine. When the stress at 5% strain was compared in the same direction, the average stress of LVL in the edge-wise direction was larger than that in timber in the tangential direction for all species, but there were no great differences between the average stress of LVL in the fl at-wise direction and that of timber in the radial direction for all species except for radiation pine. There was a close relationship between density and stress at 5% strain in LVL, especially in the edge-wise direction. For all results, radiata pine did not follow the trend of the other species; The large annual ring width of radiata pine was considered to have affected the results.     

Differences in Muscle Fiber Recruitment Patterns between Continuous and Interval Exercises

We evaluated differences in muscle fiber recruitment patterns between continuous and interval training to develop an optimal training program for Thoroughbred horses. Five well trained female thoroughbred horses (3–4 years old) were used. The horses performed two different exercises on a 10% inclined treadmill: 90%VO₂ max for 4 min (continuous) and 90% VO₂ max for 2 min × 2 times with 10-min interval (interval). Muscle samples were obtained from the middle gluteal muscle before and immediately after the exercises. Four muscle fiber types (type I, IIA, IIA/X, and IIX) were immunohistochemically identified, and the optical density of periodic acid Schiff staining (OD-PAS) in each fiber type and glycogen content of the muscle sample were determined by quantitative histochemical and biochemical procedures, respectively. No significant differences were found in the OD-PASs and glycogen contents between the continuous and interval exercises, but the decreases in OD-PAS of fast-twitch muscle fibers were obvious after interval as compared to continuous exercise. Interval exercise may be a more effective training stimulus for the glycolytic capacity of fast-twitch muscle fiber. The data about muscle fiber recruitment can provide significant insights into the optimal training program not only for thoroughbred horses, but also for human athletes.