Reinvestigation of positional distribution of tetracosahexaenoic acid in triacyl-sn-glycerols of flathead flounder flesh

Positional distribution of fatty acids in triacyl-sn-glycerols (TAG) of the flathead flounder Hippoglossoides dubius has been reinvestigated in order to accurately determine the contents of tetracosahexaenoic acid (24:6n-3) in the sn-1, sn-2, and sn-3 positions. Flesh TAG obtained from three flounders were subjected to stereospecific analysis using a suitable procedure for fish TAG analysis. The 24:6n-3 acid was found in the three positions at the concentrations of 0.3–5.5 mole% (the sn-1 position), 1.6–23.3 mole% (the sn-2 position), and 0.6–8.9 mole% (the sn-3 position). In contrast to a previous analysis, the present analysis revealed that 24:6n-3 is preferentially esterified in the sn-2 position followed by the sn-3 and sn-1 positions. Other polyunsaturated fatty acids, docosahexaenoic acid (22:6n-3; DHA) and docosapentaenoic acid (22:5n-3; DPA), showed a similar distribution pattern. These results indicate that the general tendency observed for long-chain polyunsaturated fatty acids in marine fish TAG can be extended to the distribution of 24:6n-3 in flathead flounder TAG. Because the use of flathead flounders is entirely for human food, we thus intake 24:6n-3 concentrated in the sn-2 position of their TAG.     

A new species of freshwater eel <Emphasis Type="Italic">Anguilla luzonensis</Emphasis> (Teleostei: Anguillidae) from Luzon Island of the Philippines

Anguilla luzonensis, a new species of freshwater eel, family Anguillidae, is described on the basis of 29 specimens collected from the Pinacanauan River system, which is a tributary of the Cagayan River on northern Luzon Island of the Philippines. The new species and A. celebesensis, which is distributed in the same region, both have variegated color marking and broad maxillary bands of teeth, but show statistically significant differences in the following characters of the new species: predorsal-fin length 28.6–33.1% of total length (TL); preanal length 39.6–44.8% of TL; trunk length 26.4–31.3% of TL; distance between the verticals through the anus and origin of the dorsal fin 9.3–13.9% of TL; length of gape 35.5–46.9% of head length; the total number of vertebrae 103–107; and number of abdominal vertebrae 40–42. These morphological differences, together with the present knowledge about anguillid eel taxonomy, genetics, and ecology, strongly support the presence of the new species A. luzonensis in the area of northern Luzon Island of the Philippines.     

Selenoneine,total selenium,and total mercury content in the muscle of fishes

Levels of the selenium-containing imidazole compound selenoneine and overall organic selenium were measured in the muscle of fishes by speciation analysis. The method involves monitoring ⁸²Se levels by liquid chromatography inductively coupled plasma mass spectroscopy using a gel filtration column. Selenoneine levels were found to be highest in swordfish muscle (concentration 2.8 nmol/g tissue). The selenoneine contents of bigeye tuna, Pacific bluefin tuna, albacore, yellowfin tuna, and alfonsino muscle were 1.3–2.6 nmol/g tissue. In muscle of these fishes, most organic selenium (9–42%) was present as selenoneine. In other fish species, such as Pacific sardine, greeneye, skipjack, Pacific mackerel, horse mackerel, red sea bream, and Japanese barracuda, selenoneine levels were 0.1–1.4 nmol/g tissue, accounting for 3–34% of organic selenium. In contrast, muscle of Japanese conger, Japanese anchovy, chum salmon, Pacific saury, white croaker, and marbled sole contained levels of selenoneine below the level of detection (<0.05 nmol/g tissue). Mercury and selenium contents were 0.01–5.12 nmol/g tissue and 1.4–19.1 nmol/g tissue. The Se-to-Hg molar ratio varied from species to species, ranging from 1 for swordfish to 217 for marbled sole.     

Phospholipase A<Subscript>1</Subscript> activity of crude enzyme extracted from the ovaries of skipjack tuna

The phospholipid class composition, fatty acid composition and phospholipase A₁ (PLA₁) activity from the ovaries of skipjack tuna were compared with those of six other species of marine fish. In the skipjack ovaries, the lysophosphatidylcholine (LPC) proportion for the phospholipid, the docosahexaenoic acid (DHA) percentage for the total fatty acids of the phospholipids and the PLA₁ activity of the crude enzyme were the highest among those of the seven species. The optimum pH and temperature for the PLA₁ activity of the crude enzyme from the skipjack ovaries were in the range of pH 6–7 and 20–30°C, respectively, and calcium ions were not required. As a substrate, phosphatidylcholine was more easily hydrolyzed than phosphatidylethanolamine by this enzyme, and the plasmalogen-type phospholipid was much lower than the acyl-type phospholipid. After a 6-h hydrolysis reaction of the purified phospholipid extracted from the mixed ovaries of skipjack and yellowfin tuna by this enzyme, the LPC ratio of the phospholipid increased from 20 to 72.6% and the percentage of DHA for the total fatty acids of the phospholipid also increased. Thus, skipjack ovaries might possibly be used as a source of PLA₁.     

Inter species changes of lipid compositions in liver of shallow-water sharks from the Indian Ocean

ABSTRACT:     The liver oils of five dominant shark species, Alopias superciliosus, Carcharhinus falciformis, C. longimanus, Prionace glauca , and Sphyrna lewini , in the Indian Ocean were studied for their lipid classes, fatty acid and triacylglycerol (TAG) molecular compositions, in order to understand and identify the inter species changes and possible utilization avenues. The livers showed 26.1–60.1% oil content regardless of the gender or species of shark. Triacylglycerol was the predominant lipid class (65.2–86.1%) in all species followed by sterol esters (2.9–12.7%), hydrocarbons (2.5–11.6%) and diacylglyceryl ethers (1.0–3.8%). The major fatty acid in total lipid and TAG was docosahexaenoic acid (C22:6n-3; DHA), which in male S. lewini was at remarkable levels (43.4%). The predominant fatty acid combinations of TAG molecules in all shark species were palmitic, DHA and oleic acids (PDO 12.5–19.9%), and palmitic, DHA and palmitic acids (PDP; 8.4–15.4%). Tridocosahexaenoin was found in considerable levels in S. lewini (male).     

Changes in the scotopic vision of juvenile Pacific bluefin tuna (Thunnus orientalis) with growth

In cultured juvenile Pacific bluefin tuna (Thunnus orientalis), reducing the mass deaths caused by collision or contact with tank or net walls at night is a priority for seedling production. Pacific bluefin tuna is a visually dependant species, although its scotopic vision is poor. We recorded electroretinograms to investigate the visual function with growth in the dark-adapted eyes of juvenile Pacific bluefin tuna. Peak wavelengths of spectral sensitivity [38–62 days posthatch (dph), 77–167 mm standard length (SL)] were observed between 474 and 494 nm. Visual light sensitivity has a tendency to increase slightly with growth at 28–64 dph in individuals that measured 29–175 mm SL. However, visual temporal resolution did not significantly increase with growth at 38–62 days dph in individuals that measured 77–167 mm SL. These results suggest that the mass death continues between 28 and 64 dph because of low visual function and increasing swimming speed with growth.     

实用饲料添加肌醇对草鱼生长、脂质代谢及抗氧化机能的影响

为探讨肌醇对草鱼生长、脂质代谢及抗氧化机能的影响,以实用饲料配方为基础,分别添加0(对照)、50、100、150、200、300和400 mg/kg肌醇,配制成7组等氮等脂的饲料,每组饲料设4个重复,每个重复饲喂初始体质量为(15.00±0.15)g的草鱼25尾,养殖56 d。结果显示,饲料中添加100~150 mg/kg肌醇能显著提高草鱼终末均重(FBW)、增重率(WGR)、特定生长率(SGR)及饲料效率(FE);随饲料肌醇添加水平的上升,全鱼脂肪、肝脏脂肪和脂肪沉积率先升后降,在50~300 mg/kg均与对照组存在显著差异,且均在100 mg/kg达到最大值。肌肉脂肪则逐渐下降并趋于稳定,在100 mg/kg达到最小值,100~400 mg/kg差异不显著;肠脂肪酶、血浆总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)含量及高密度/低密度脂蛋白胆固醇(HDL-C/LDL-C)均呈先升后降的趋势,除LDL-C在各组间差异不显著外,其余指标均在100~150 mg/kg达到最大值;添加肌醇能显著增强肝脏和肌肉中肉碱脂酰转移酶(CPT-I)和乙酰辅酶A羧化酶(ACC)的活性。与对照组相比,100 mg/kg肝脏CPT-I的增幅比例低于ACC的增幅比例,肌肉则相反;当肌醇添加水平为100~150 mg/kg时,肝脏和肌肉中超氧化物歧化酶(SOD)的活性显著升高,丙二醛(MDA)含量及血浆中谷丙转氨酶(ALT)、谷草转氨酶(AST)均显著降低。研究表明,实用饲料添加适宜的肌醇能改善草鱼的生长、饲料转化和肝脏功能,促进脂肪消化,加快脂肪酸的合成与分解代谢,使全鱼和肝脏增脂、肌肉降脂,且能够提高肝脏和肌肉的抗氧化机能。以FE和SGR为效应指标,草鱼实用饲料肌醇适宜添加量为90.3~96.4 mg/kg。     

Lipids in female northern bluefin tuna (Thunnus thynnus thynnus L.) during sexual maturation

Total lipids, lipid classes, fatty acids and vitamin E contents were measured in ovary, liver, dark muscle, white muscle and mesenteric perigonadal fat in four maturational stages of Atlantic female northern buefin tuna (Thunnus thynnus thynnus L.) during migration to Mediterranean spawning grounds through the Strait of Gibraltar. Ovary lipid content increased significantly by 37.6-fold from inmature to spawning stage, accumulating large quantities of steryl/wax esters, whereas the ratio of mesenteric perigonadal fat (primarily triacylglycerols) weight to ovary weight decreased linearly throughout maturation. In contrast, liver, dark red and white muscles lipid content did not show a major depletion during maturation compared to that of perigonadal fat. The fatty acid composition of ovary from bluefin tuna showed that docosahexaenoic acid (DHA; 22:6n-3) was the most abundant fatty acid and DHA: EPA: AA and DHA: EPA ratios were 19.3/4.3/1 and 4.5/1, respectively. Neutral lipids predominated over polar lipids (2-3:1) in ovary and triacylglycerol and steryl/wax ester were major lipid classes. In general, fatty acid composition of most tissues examined were similar to each other but, in contrast, the fatty acid profile of perigonadal fat was rich in monoenes such as 18:1n-9, 20:1n-9 and 22:1n-11, and DHA decreased significantly at spawning stage. The total lipid balance suggested for the disappearance of lipid from perigonadal fat depots and, to a lesser degree, from muscle and liver depots, reflected lipid utilization for gonadal development on the one hand and non-gonadal metabolism in the other. The former included lipid transferred to the gonad and lipid catabolized to provide metabolic energy for the biosynthesis of gonadal constituents and the later was likely to be lipid catabolism to provide energy for swimming during spawning migration. This revised version was published online in August 2006 with corrections to the Cover Date.     

Influence of Varying Dietary Protein Content at Three Lipid Concentrations on Growth Characteristics of Juvenile Swordtails (Xiphophorus helleri Heckel 1848)

Three protein levels (30%, 38% and 45%) at three different dietary lipid concentrations (6%, 8% and 12%) were used to formulate nine different diets that were fed for 60 days to 6–8 weeks old juvenile swordtails (Xiphophorus helleri). It would appear that a diet of at least 45% protein at a 6% lipid concentration is needed for the best specific growth rate (1.27% per day) and feed conversion ratio (2.50) at this specific growth phase in X. helleri. No ‘protein sparing effect’ was found when lipid concentration was increased at lower protein levels. This revised version was published online in August 2006 with corrections to the Cover Date.     

Influence of the gametogenic cycle on the biochemical composition of the ovary of the great scallop

The oogenic cycle and biochemical composition of the ovary of raft-cultured great scallop (Pecten maximus) were studied during the period April 1990–July 1991. The ovary condition index (FGI) and stereological studies showed the existence of two principal spawning periods, winter and late spring-early summer. No sexual resting period was found. Oocyte lysis was high throughout the year. Ovarian lipid levels displayed a clear seasonal pattern linked to the gametogenic cycle. Total lipid (TL, 16–21% dry weight), acylglycerol (AG, 20–65% TL) and free sterol (FS, 2.8–6.4% TL) levels were, generally, higher in the ripe ovary and a decrease coincided with spawning. Protein (59–63% dry weight), glycogen (<3% dry weight) and phospholipid (PL, 26–35% TL) levels showed no clear seasonal trend. The TL and AG were a good index of ovarian sexual maturity. The TL % of dry weight) correlated well with the female gonad condition index (r _s = 0.779, p < 0.001), and AG (% f TL) correlated well with the mean oocyte diameter (r _s = 0.630, p < 0.01) and the female gonad condition index (r _s = 0.443, p < 0.05).     

Annual variation of potential predation impacts on larval and juvenile marbled sole <Emphasis Type="Italic">Pseudopleuronectes yokohamae</Emphasis> by sand shrimp <Emphasis Type="Italic">Crangon uritai</Emphasis> in Hakodate Bay,Hokkaido

An investigation was conducted to evaluate the annual variation in potential predation impact (PPI) on larval and juvenile Pseudopleuronectes yokohamae by Crangon uritai in Hakodate Bay using predator-prey size relationships. Laboratory experiments were designed to estimate the favorable prey size of C. uritai through back-calculation of body length (BL) of P. yokohamae from sagittal otolith diameter observed in the stomachs of C. uritai. The most favorable prey-predator size ratio (BL of P. yokohamae-total length (TL) of C. uritai) class was 0.15–0.19, and ranged from 0.12–0.31. There was a significant positive correlation between the BL increase of P. yokohamae and the bottom water temperature in the field, such that BLs stagnated during the cold year of 1999 from April to June, and increased during the warm year of 2002. In contrast, no significant correlation was found between the TL increase of C. uritai and the bottom water temperature. Moreover, there were no significant differences in the mean TL of C. uritai between months (April–June) or years (1998–2002). Variation of PPI depended on the relationships between the growth rates of P. yokohamae and bottom water temperature. Therefore, the warm year of 2002 resulted in higher potential predation impact (PPI≥20), and it was at least 20 days shorter than that of the cold year of 1999. These results suggest that increased bottom water temperature in the nursery area was one of the most important factors for cumulative predation loss.     

Effects of open- and closed-system temperature changes on blood O2-binding characteristics of Atlantic bluefin tuna (Thunnus thynnus)

We investigated the effects of open- and closed-system temperature changes on the O₂ affinity of Atlantic bluefin tuna (Thunnus thynnus) blood using in vitro methods essentially identical to those previously employed on tropical tuna species. Bluefin tuna blood has a general O₂ affinity (P ₅₀ = 2.6–3.1 kPa or 19–23 mm Hg at 0.5% CO₂) similar to that of skipjack tuna, yellowfin tuna, and kawakawa blood (P ₅₀ = 2.8–3.1 kPa at 0.5% CO₂) but significantly above that of bigeye tuna blood (P ₅₀ = 1.6–2.0 kPa at 0.5% CO₂). We therefore hypothesize that bluefin tuna are less tolerant of hypoxia than bigeye tuna. Further, we found the P ₅₀ of bluefin tuna blood to be slightly reduced by a 10°C open-system temperature increase (e.g., from 4.83 kPa at 15°C to 3.95 kPa at 25°C) and to be completely unaffected by a 10°C closed-system temperature change. Bluefin tuna blood, therefore, had a significantly reduced Bohr effect when subjected to the inevitable changes in P CO ₂ and plasma pH that accompany closed-system temperature shifts (0.04–0.09 Δlog P₅₀ΔpH⁻¹) compared with the effects of changes in plasma pH accomplished by changing P CO ₂ alone (0.81–0.94 Δlog P₅₀ Δ pH⁻¹). This response is similar to that of skipjack tuna blood, but different from yellowfin or bigeye tuna blood. During closed-system temperature changes at oxygen levels above P ₅₀, however, bluefin tuna blood showed a reversed temperature effect (i.e., P O ₂ decreased in response to an increase in temperature). Unlike in other tuna species, temperature effects on O₂ affinity of bluefin tuna whole blood were similar to those previously reported for hemoglobin solutions, suggesting that red cell-mediated ligand changes are not involved.     

Lipid classes from marine species and meals intended for cephalopod feeding

In this study, HPTLC was used to make a quantitative analysis of the total lipid content (TL dry weight) and their classes in 39 samples, including molluscs, crustaceans, fish and meals in an attempt to identify those most suitable for formulating diets for cephalopods by reference to an index of nutritional quality (OI_LC: Oser’s index modified for lipid classes). All the crustaceans analysed (<10% TL), fish from artisanal fisheries such as Boops boops, Gadus poutassou, Mugil sp. and Gadus minutus (<10% TL) and the plant meals (TL < 5%) would cause fewer digestive problems than the fish that were a by-catch from fish farms (B. boops or Sparus aurata), pelagic species (Sardina pilchardus or Trachurus trachurus) or krill meal, which are characterised by their high lipid content (20–60% TL). These latter feeds were associated with neutral lipids, mainly triglycerides during the summer. Mytilus galloprovincialis, Carcinus maenas, G. poutassou, Mugil sp., S. pilchardus and G. minutus had a more appropriate lipid content and profile during the winter, when they showed a higher OI_LC due to the greater variety of polar lipid classes they contained. Phospholipids like PS, PI and PE seem to be limiting nutrients in cephalopods because of their high content (78–542, 41–309 and 152–2,114 mg/100 g, respectively) compared with the rest of the samples. None of the meals analysed showed a good nutritional balance per se and should only be used in conjunction with other foods.     

The effect of docosahexaenoic and eicosapentaenoic acids in live food on the development of abnormal morphology in hatchery-reared brown sole Pseudopleuronectes herzensteini

We examined the effect of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) on the rates of abnormal morphology in juvenile brown sole Pseudopleuronectes herzensteini. Larvae during the D–E stages (15–24 days post hatching) were fed live food containing various amounts of DHA and/or EPA prepared using emulsified oils (DHA ethyl ester, EPA ethyl ester, and corn oil). Larvae during the F–I stages were fed Artemia enriched with a commercial diet supplement. We found that DHA and EPA promoted larval development and improved the incidence of morphological abnormalities in brown sole juveniles to a similar extent. However, DHA was more effective than EPA in preventing the appearance of morphological abnormalities in brown sole. The incidence of normal morphology was clearly improved by an increase of the DHA content in brown sole larvae at 25 days post-hatching. These results suggest that it is important to promote larval development and feed larvae with live food containing high levels of DHA during the D–E stages to prevent morphological abnormalities in brown sole juveniles.     

Effects of dietary lipid levels on growth performance,feed utilization and fatty acid composition of juvenile Japanese seabass (<Emphasis Type="Italic">Lateolabrax japonicus</Emphasis>) reared in seawater

Triplicate groups of 40 Japanese seabass Lateolabrax japonicus (initial weight, 11.3 ± 0.4 g) reared in seawater (salinity, 30.0–33.0 g L⁻¹) were fed with five isonitrogenous (41.3 ± 0.2% crude protein) and isoenergetic (18.5 ± 0.3 MJ kg⁻¹) experimental diets formulated with increasing lipid levels (4.3, 8.4, 12.2, 15.8 and 20.1% lipid) for 10 weeks. Survival throughout the feeding experiment ranged from 87.5 to 100%, but the survival of fingerlings fed with the 4.3% lipid diet was significantly lower than the rest of the diets. At the end of the feeding experiment, fish fed with 12.2% lipid diet showed optimal growth performance (P < 0.05). Lipid contents of whole body, liver and muscle increased in parallel with the increase in dietary lipid levels. Viscerosomatic index (VSI), hepatosomatic index (HSI) and muscle lipid content were higher in 20.1% lipid group than those in the rest of the lipid level groups indicating that viscera and muscle tissues played important contributions to body lipid deposition. High proportions of 18:1n-9, eicosapentaenoic acid (20:5n-3; EPA) and docosahexaenoic acid (22:6n-3; DHA), and low concentrations of n-6 polyunsaturated fatty acids (PUFAs) occurring in liver and muscle, to some extent, reflected fatty acid (FAs) composition in the experimental diets.     

Effects of different diets on juvenile tench, Tinca tinca (L.) reared under controlled conditions

In a laboratory experiment at 28 °C, four commercial starters (Carp Starter, Uni Starter, Perla Plus, Asta) were fed intensively to 11-month-old juvenile tench Tinca tinca of initially 41 mm TL and 0.70 g BW, over a period of 120 days. Frozen chironomid larvae were fed to fish below satiation as a reference diet for comparison of the fish body chemical composition. Final survival rates of 99–100% and satisfactory growth were attained with all dry diets, ranging from 73.3 (Carp Starter) to 84.8 mm TL (Asta) and from 5.39 (Carp Starter) to 8.09 g BW (Perla Plus). External body deformities of up to 96.4% specimens appeared in those fed Uni Starter, Carp Starter and Perla Plus, whereas 0% were recorded for Asta and chironomids. Fish in groups with deformities showed significantly elevated lipid content in their bodies, lower ash content and higher condition coefficient values. Juveniles in groups without deformities did not substantially change lipid content, but the ash content increased. Thus, among four dry diets studied, only Asta starter can be fed effectively and safely to juvenile tench reared intensively under controlled conditions.     

The effects of dietary lipid sources and lecithin on the production of giant freshwater prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis> larvae in the Mekong Delta region of Vietnam

The giant freshwater prawn, Macrobrachium rosenbergii, is cultured widely in the Mekong Delta region of Vietnam but it is often difficult or expensive for hatchery operators to purchase commercial diets used as a feeding supplement to Artemia nauplii. Therefore, in the present study, the effects of lipid sources and lecithin on the growth and survival rate of M. rosenbergii larvae were examined in order to develop suitable hand-prepared larval diets for seed production of M. rosenbergii in this area. Six egg custard diets consisting of various ratios of lipid (originating from soybean oil and squid oil) and lecithin were used for rearing Macrobrachium rosenbergii larvae. Treatments in which larvae were fed diets containing squid oil exhibited the highest body length and survival rates (7.14–7.43 mm and 51.1–68.1%, respectively), and differed significantly from other treatments (P<0.05). Use of dietary soybean oil yielded the lowest body length and survival rates (6.29–6.75 mm and 22.0–48.7%), respectively). The supplementation of dietary lecithin did not increase final body weight but did improve larval survival rates. The n-3 HUFA content of prawns fed dietary squid oil was higher than those of animals provided with other diets. These results indicated that the most appropriate diet for rearing M. rosenbergii larvae is the diet containing 3% squid oil and 1.5% lecithin.     

Myxosporean and microsporidian infections in cultured Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> in Japan

During the parasitological survey of cultured juvenile Pacific bluefin tuna (PBT) Thunnus orientalis in 2007 and 2008, two myxosporeans and one microsporidian were found. Morphological and molecular analysis showed that the heart-infecting and brain-infecting myxosporeans are identified as Kudoa shiomitsui and K. yasunagai, respectively. This is a new host record for both species. High prevalence of infection (77–100%) with K. shiomitsui was observed in October to December (1–2 months post-transfer to sea cages), whereas only a few fish were infected with K. yasunagai. A microsporidian observed as white “cysts” in the trunk muscle of PBT had a resemblance to Microsporidium seriolae, the causative parasite of beko disease in yellowtail. Phylogenetic analysis showed that the microsporidian from PBT is closely related to but distinct from several other muscle-infecting species such as M. seriolae, Microsporidium sp. RSB, and Microsporidium sp. SH. Additionally, the spore dimension (2.7 × 1.5 μm on average) was remarkably smaller than the others. These results suggest that the present microsporidian is an undescribed species and designated Microsporidium sp. PBT. Prevalence and intensity of infection with Microsporidium sp. PBT reached a maximum of 100% and 20 cysts/fish, respectively. Although pathogenic effects of the two Kudoa species on fish health remain unknown, the microsporidian could be of concern to PBT aquaculture due to unsightly cysts in the musculature, reducing the market value of the fish.     

Effect of ration size on growth,body composition,and energy and protein maintenance requirement of fingerling Indian major carp, <Emphasis Type="Italic">Labeo rohita</Emphasis> (Hamilton)

An eight-week feeding trial has been conducted to determine the optimum ration for Indian major carp, Labeo rohita, fingerling (4.10 ± 0.30 cm, 0.55 ± 0.16 g) by feeding a purified diet (40% CP; 3.61 kcal g⁻¹ GE) at six levels, 2, 4, 6, 8, 10, and 12% of body weight per day, at 0800 and 1600 h, in triplicate, to 20 fish per trough fitted with a water flow-through system. Highest weight gain, best feed conversion ratio (FCR), best specific growth rate (SGR%), and highest protein efficiency ratio (PER) were evident for rations of 6–8% body weight. Second-degree polynomial regression analysis for FCR, PER, protein, and energy retention data indicated the break-points occurred at 6.55, 6.75, 6.80, and 6.95% bw per day, respectively. Significant (P < 0.05) differences between body composition were observed for fish fed different rations. Maximum body protein content was recorded for 6% and 8% rations. A linear increase in body fat content was evident with increasing ration. Body moisture and ash content remained non-significantly (P > 0.05) low for higher rations, however. On the basis of these results it is recommended that feeding in the range 6.5–7.0% bw per day corresponding to 2.6–2.8 g protein and 23.49–25.31 kcal energy per 100 g of the diet per day is optimum for growth and efficient feed utilization of Labeo rohita. Results for 2–4% rations (0.8–1.6 g protein and 7.23–14.46 kcal energy) suggest these amounts approximate to the maintenance requirement of this fish.