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.     

Temporal expression analyses of pancreatic and gastric digestive enzymes during early development of the olive flounder (Paralichthys olivaceus)

The olive flounder (Paralichthys olivaceus) is one of the most important aquaculture species in Korea. Previous studies focused on the morphological changes and only analysed digestive enzyme expression patterns of certain digestive systems. A comprehensive analysis of the pancreatic and gastric digestive enzymes of the olive flounder in its early developmental stages has not been performed. In this study, we determined the mRNA expression levels of pancreas and stomach digestive enzymes by extracting total RNA from the whole body of olive flounders from hatching to post‐metamorphosis and conducting real‐time PCR analysis with gene‐specific primers for each isoform. The pancreatic digestive enzymes were initially and strongly expressed from the first feeding period to pre‐metamorphosis. The expression of trypsinogen 3 was detected from the early stage of development to metamorphosis, whereas trypsinogen 2 was significantly increased only in the post‐metamorphosis period. The mRNA expression of pepsinogen was only detected at metamorphosis and corresponded to stomach differentiation. The lipid digestive‐related enzymes had already reached a certain level at the beginning of hatching, then increased during the early developmental stage and gradually decreased before metamorphosis. These results suggest that food ingestion is exclusively digested by pancreatic digestive enzymes and trypsinogen 3 during the early developmental stage until pre‐metamorphosis, and it then fully switches to digestion by pepsinogen and trypsinogen 2 in mid‐metamorphosis. In conclusion, understanding the expression profiles of digestive enzymes in the pancreas and stomach during the early developmental stages is an important consideration when developing formulated larval diets for aquacultured olive flounder.     

Application of microparticle diets for Japanese flounder Paralichthys olivaceus larvae

ABSTRACT:   A feeding trial was conducted with two newly developed microparticle diets (MD-Q and MD-V), which differed in the mixing ratios of two types of casein hydrolysates (C700 and C800) as the protein source. The ratio of C700 and C800 were adjusted to 7:3 in MD-Q and 6:4 in MD-V, respectively. Japanese flounder larvae were fed from 7 days after hatching (d.a.h.) to 32 d.a.h. on live food (LF), MD solely, MD + 1/3 LF (either of the MD and one-third quantity of the live food) and 1/3 LF (one-third quantity of live food alone), respectively. They were then switched to Kyowa diet B from 33 d.a.h. until 40 d.a.h. The larvae fed on MD-Q exclusively had a higher survival rate (36.4%) than those fed on MD-V (24.2%) by 22 d.a.h. Unfed larvae could survive up to only 12 d.a.h. Larvae fed on MD-Q + 1/3 LF also had a significantly higher survival rate (34.2%) than those on MD-V + 1/3 LF (16.3%) and 1/3 LF solely (15.4%) at 32 d.a.h. These results suggest the potential of MD-Q as a part replacement for LF from the early developmental stage in the seed production of Japanese flounder.     

Development of microparticle diets for Japanese flounder Paralichthys olivaceus larvae

ABSTRACT:   Two newly designed microparticle diets (MD), with two kinds of peptide (C700 and C800) as a protein source were developed. Microparticle diet Q (MD-Q) contained C700 (molecular weight ∼30 000 Da) and C800 (molecular weight 1000–2000 Da); in contrast, microparticle diet T (MD-T) contained C700 only. Two separate trials, representing larvae from different hatches, were conducted. Japanese flounder Paralichthys olivaceus larvae were fed newly designed MD or a combination of them with live food (LF) between 3 and 10 days after hatching (d.a.h.) in experiment I, or between 11 and 20 d.a.h. in experiment II, and compared them to LF and a commercial diet (CD) feeding groups. The growth and survival rates in both experiments were the highest in the LF treatment. But in the microparticle treatment larvae fed MD-Q had a higher survival rate and better growth than those fed MD-T, either alone or in combination with LF. First-feeding larvae fed on MD-Q had a 20.5% survival rate and 1.12 mm gain by 10 d.a.h. Unfed larvae died within 6 d.a.h. These results indicate that using a mixture of different molecular weight peptides is good protein sources and also this type of microparticle diet can be applied to flounder from larval to juvenile stages.     

Ontogeny of pancreatic enzymes in larval red drum Sciaenops ocellatus

The growth, survival and trypsin, lipase and amylase activities of red drum larvae were measured in two experiments. For the first trial, a group was fed live prey only (L) and another group was fed a combination of a microparticulate diet (MPD) and live food (L-MP). For the second growth trial a group fed the MPD only (MP) and a starvation group (ST) were examined in addition to the L and L-MP treatments. Enzyme activities of live prey were measured to estimate their possible contribution to larval digestion. No significant ( P  > 0.05) differences in final size and survival were observed between treatments L and L-MP. Larvae subjected to starvation or fed the MPD diet alone were smaller than treatments fed live prey and did not survive past days 5 and 14, respectively. Trypsin, lipase and amylase activities were detectable at hatching. No significant differences ( P  > 0.05) in total enzyme activities among treatments were observed before day 14. Specific activity of trypsin, lipase and amylase peaked on day 3 (prior to first feeding) and subsequently decreased. For trypsin, the percentage of enzyme activity potentially attributable to ingested prey increased with age to a maximum of 17%. For lipase and amylase this fraction was less than 5% throughout the study, except on day 8 (12% and 24%, respectively). The lack of significant differences observed in the activity of digestive enzymes among treatments suggests that dietary regime, availability of prey and possible effects of exogenous enzymes did not significantly influence enzyme activity. Therefore, the lower growth rate observed in the L-MP, MP and starved treatments cannot be attributed to low digestive enzyme production of the enzymes measured. It is more likely that the MPD failed to supply the required nutrients for adequate development.     

Effect of dietary taurine and lipid contents on conjugated bile acid composition and growth performance of juvenile Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

ABSTRACT:   The effects of dietary taurine levels and lipid contents on the conjugated bile acid composition and growth performance of juvenile Japanese flounder Paralichthys olivaceus were investigated. Six types of diet (three different levels of taurine at two different levels of lipid) were fed to juveniles (average body weight, 0.04 g). Fishmeal that was washed with 70% ethanol to remove taurine was used as the sole protein source. Feeding experiments were carried out at 20°C for 6 weeks. At the end of the experiments, fish were weighed and analyzed for free amino acids in the body and the composition of the conjugated bile acids. The body weight and percent weight gain of the juveniles were improved by the dietary taurine supplementation. The taurine contents of the whole body and tissues increased with the increase of the dietary taurine contents. The conjugated bile acids in the gall bladder consisted of taurocholic acid and taurochenodeoxycholic acid, which increased with the increase of the dietary taurine level. Taurocholic acid accounted for more than 95% of the total conjugated bile acids. This indicates that taurine is the sole amino acid to conjugate bile acid in Japanese flounder.     

克氏原螯虾幼体发育时期消化酶活力及氨基酸含量研究

采用生物化学方法对克氏原螯虾各期幼体消化酶活力与氨基酸组成进行了分析。实验结果显示,在克氏原螯虾幼体发育过程中,五种消化酶活力(胃蛋白酶、类胰蛋白酶、淀粉酶、纤维素酶和脂肪酶)表现出两种变化模式:胃蛋白酶和类胰蛋白酶在幼体发育Ⅱ龄幼体期和Ⅳ龄幼体期活力较高,其中Ⅳ龄幼体期该两种酶活力最高;淀粉酶、纤维素酶和脂肪酶活力为先增高后降低,在Ⅲ龄幼体期,三种酶的活力达到最高,但与Ⅳ龄幼体期的酶活力比较无显著差异。总氨基酸含量在幼体发育早期逐渐降低,幼体发育至Ⅲ龄幼体期,其氨基酸含量最低,Ⅳ龄幼体期又有所增加。在测定的所有氨基酸中谷氨酸含量最高,必需氨基酸中赖氨酸含量最高,单个必需氨基酸含量与必需氨基酸总量的比值(A/E)在整个胚胎发育过程中的变化趋于一致。     

兴凯湖红鳍原鲔消化酶活力和肌肉氨基酸组成的季节变化

研究了不同季节体质量为107．824-22．97g的兴凯湖野生红鳍原鲐（Cultrichthys erythropterus）肝胰脏、肠道消化酶活力及夏季昼夜变化和肌肉氨基酸种类组成、含量及其变化,探讨其营养生态特性及变化规律。结果显示：不同季节红鳍原铂的蛋白酶、淀粉酶和脂肪酶的比活性显著不同（P〈0．05）,夏、秋季高于春、冬季,同一季节蛋白酶比活均显著高于淀粉酶比活（P〈0．05）;红鳍原鲐蛋白酶、淀粉酶和脂肪酶比活的昼夜变化为：19：00～21：00、19：00和16：00～19：00时值最高,7：00、7：00和21：00值最低。红鳍原鲴肌肉由18种氨基酸组成,冬季缬氨酸、蛋氨酸、异亮氨酸、苯丙氨酸和赖氨酸五种必需氨基酸的含量显著低于春、秋季（P〈0．05）,而非必需氨基酸的含量无明显的季节变化;不同季节谷氨酸含量均最高,最低为胱氨酸。春、夏季氨基酸总量和必需氨基酸总量较高,秋、冬季较低;秋季鲜味氨基酸含量最低。研究表明,野生红鳍原鲐消化生理具有不同季节和日变化特点,氨基酸组成的季节变化提示：冬季鱼体代谢活动对肌肉组织中必需氨基酸的依赖。