Tsuyoshi Sugita

Supplemental effect of bile salts to soybean meal-based diet on growth and feed utilization of rainbow trout Oncorhynchus mykiss

A feeding experiment was conducted to evaluate the supplemental effect of bile salts to a defatted soybean meal-based non-fish meal diet for rainbow trout Oncorhynchus mykiss. A fish meal-based diet (FM) and two non-fish meal diets with and without bovine bile salts (SC+B and SC, respectively) were fed to fish (13 g initial weight) for 10 weeks. Fish fed diet SC showed inferior growth and feed efficiency, while bile salt supplementation improved the parameters to the same levels as fish fed diet FM. Crude fat and starch digestibility of diet SC-fed fish decreased after the 10-week feeding trial compared to the data obtained with fish that had no experience of the diet. Total biliary bile salt content and intestinal maltase activity of fish fed diet SC were the lowest among treatments, while these parameters were improved by bile salt supplementation. Morphological changes occurred in the distal intestine and liver of the diet SC group, although the histological features of fish fed diet SC+B were similar to those of fish fed diet FM. These results suggest that bile salt supplementation to a soybean meal-based diet improves the nutrient utilization by normalizing digestive processes in rainbow trout.

Influence of certain soybean antinutritional factors supplemented to a casein-based semipurified diet on intestinal and liver morphology in fingerling rainbow trout Oncorhynchus mykiss

To investigate the cause of morphological changes occurring in the liver and intestine of rainbow trout Oncorhynchus mykiss fed defatted soybean meal (SBM)-based non-fish meal diets, morphological observations were conducted on fish fed casein-based semipurified diets supplemented with soybean antinutritional factors (soya saponin, soya isoflavone, soya lectin, oligosaccharide, Ca-phytate and their mixture). Hepatocytes and the epithelial cells in the hepatic ducts were not atrophied in any of the treatments. Lack of microvilli and pinocytotic vacuoles, and fatty degeneration of the epithelial cells were observed in the distal intestine of fish fed diets containing soya saponin. The proliferation of connective tissue in the mucosal folds and submucosa was shown in the distal intestine of fish fed the diet containing all substances including lectin. These results suggest that both saponin and lectin are the responsible factors in SBM for the occurrence of morphological changes of the distal intestine in rainbow trout fed SBM-based diets.

Supplemental effect of cholyltaurine and soybean lecithin to a soybean meal-based fish meal-free diet on hepatic and intestinal morphology of rainbow trout Oncorhynchus mykiss

The supplemental effects of cholyltaurine and soybean lecithin to a defatted soybean meal (SBM)-based fish meal-free diet on the hepatic and intestinal morphology of rainbow trout Oncorhynchus mykiss were investigated. A fish meal-based diet (FM), three fish meal-free diets supplemented either with soybean lecithin (SL), bovine gall powder (BG) or cholyltaurine (C-tau) and an unsupplemented fish meal-free diet (NFM), were given to fish (−11 g) for 10 weeks. The growth was the lowest in fish fed diet NFM, intermediate in fish fed diet SL and the highest in fish fed diet FM, BG and C-tau. Hepatocytes of fish fed diet NFM were atrophied, and the distal intestine of these fish showed abnormal features: disintegrated microvilli and fatty degeneration in epithelial cells and increased amount of connective tissue in the submucosa. Hepatic and intestinal histological features in fish fed diets SL, BG and C-tau were similar to those in fish fed diet FM. These results indicate that soybean lecithin, bovine bile salts and cholyltaurine have similar effects on normalizing the hepatic and intestinal morphologies of rainbow trout fed the SBM-based diet, although growth promotion effect was limited in the soybean lecithin.

Effects of feeding time, water temperature, feeding frequency and dietary composition on apparent nutrient digestibility in rainbow trout Oncorhynchus mykiss and common carp Cyprinus carpio

A series of experiments was conducted to evaluate the effects of feeding time (daytime vs nighttime feeding), dietary fat content (8 and 20%), feeding frequency (trout 1–3 times/day, carp 2–7 times/day), water temperature (trout 18 and 11°C, carp 25 and 17°C), on the apparent nutrient digestibility in rainbow trout and common carp. The feeding time had little effect on the macronutrient digestibility in both species. In trout, starch digestibility decreased with the decrease of water temperature and with increase of feeding frequency, but in carp, increase of the feeding frequency markedly decreased the macronutrient digestibility and phosphrrus absorption of the high fat diet. Fat digestibility of the beef tallow diet decreased relative to the pollock oil diet in carp, without affecting the phosphorus absorption. Inclusion of raw starch, the digestibility of which was lower than that of gelatinized starch, increased the phosphorus absorption in carp. These results suggest that reduction of water temperature and increase of feeding frequency notably decreased starch digestibility in trout while in carp, the effects of feeding frequency and water temperature on macronutrient digestibility and phosphorus absorption are notable for a high fat diet.

Response of enzyme activities and metabolic intermediate concentrations to a long burst of exercise and following resting in muscle and the hepatopancreas of carp

The authors are investigating the regulation of carbohydrate metabolism in fish mainly from the aspect of enzymes. In previous studies, we reported that the activities of carbohydrate-metabolizing enzymes responded slowly to feeding rates, feed composition, acclimation temperature, and so on, suggesting an induction of enzyme proteins. Moreover, it was suggested that phosphofructokinase (PFK) plays an important role in glycolysis. Thus, although long-term responses of carbohydrate-metabolizing enzymes become remarkably clear, the short-term response of carbohydrate metabolism is still unknown. To elucidate the short-time response of carbohydrate metabolism in fish, the present study investigated the response of enzyme activities and metabolic intermediate concentrations to short-time exercise, and following resting in the muscle and hepatopancreas of carp.

Effects of biotechnologically processed soybean meals in a nonfishmeal diet on growth performance, bile acid status, and morphological condition of the distal intestine and liver of rainbow trout Oncorhynchus mykiss

This study evaluated four commercially available biotechnologically processed soybean meals (SBMs) that were included as the primary protein sources in a nonfishmeal diet for juvenile rainbow trout Oncorhynchus mykiss. The processed SBMs were Afcep SBM fermented with mixed bacteria predominantly Bacillus spp., DaBomb SBM fermented with Lactobacillus acidophilus, PepSoyGen SBM fermented with Aspergillus oryzae and Bacillus subtilis, and HP300 SBM treated enzymatically. Fishmeal (FM) and common heat-treated and hulled SBM were used in the reference diets. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) analysis revealed that the protein components of Afcep SBM appeared to have relatively small molecular sizes compared with the other SBM products. After a 10-week feeding trial, growth of fish fed the common SBM diet was depressed, and abnormal liver and distal intestinal morphologies and bile acid characteristics were observed. The Afcep SBM diet showed approximated growth performance and comparable physiological conditions relative to the FM diet, while only limited improvements were observed in fish fed the other processed SBMs. These findings suggest that Afcep SBM, with its smaller molecular mass proteins, is the most promising processed SBM in nonfishmeal diets for rainbow trout.

Postprandial response and tissue distribution of the bile acid synthesis-related genes, cyp7a1, cyp8b1 and shp, in rainbow trout Oncorhynchus mykiss

In mammals, cholesterol 7α-hydroxylase (CYP7A1) and sterol 12α-hydroxylase (CYP8B1) are rate-limiting enzymes in bile acid synthesis. In addition, a small heterodimer partner (SHP) is also known to inhibit bile acid synthesis via the suppression of CYP7A1 and CYP8B1 expression. However, little information is currently available regarding primary structure of the genes involved in bile acid synthesis in fish. We therefore cloned cyp7a1, cyp8b1 and shp genes from rainbow trout and obtained cDNAs encoding two isoforms each of Cyp7a1 (-1 and -2), Cyp8b1 (-1 and -2) and Shp (-1 and -2). Both cyp7a1-1 and -2 encoded proteins of 512 amino acids. Trout cyp7a1-1 was expressed not only primarily in the kidney, pyloric caecum and mid-gut, but also weakly in the liver, eye, gill and ovary. cyp7a1-2 was highly expressed in the liver, pyloric caecum and mid-gut. cyp8b1-1 and -2, which encoded proteins of 512 and 509 amino acids, respectively, were principally expressed in the liver. Both shp-1 and -2, which encoded proteins of 288 and 290 amino acids, respectively, were strongly expressed in the liver, but shp-2 was also highly expressed in the gallbladder and digestive tract. The temporal changes in the expression of cyp7a1-1/-2, cyp8b1-1/-2 and shp-1/-2 in the liver were assessed after consumption of a single meal. Expression of cyp7a1-1/-2 and cyp8b1-1/-2 increased within 3h post feeding (hpf) when the stomach was still approximately 84% full and the gallbladder was almost completely empty. Although the expression of shp-1 did not change after feeding, the expression pattern of shp-2 was inversely related to the expression patterns of cyp7a1-1/-2 and cyp8b1-1/-2. Specifically, shp-2 expression decreased until 3 hpf before returning to initial levels at 24 hpf. These findings suggest that Cyp7a1s/8b1s and Shp-2 function antagonistically in bile acid synthesis in rainbow trout.