A simple test to estimate the salinity resistance of fish with specific application to O. niloticus and S. melanotheron

In order to develop a simple and accurate index of the salinity resistance of tilapia, batches of 10 juveniles (5 to 20 g) of two different species Oreochromis niloticus and Sarotherodon melanotheron reared in freshwater were subjected to gradual increases in salinity until 100% mortality. Seven daily increments of salinity were tested with 4 replicates: 2, 4, 6, 8, 10, 12 and 14 g l⁻¹ day⁻¹, while control batches were kept in fresh water. The temperature was maintained at 27 °C. The concentration of oxygen, ammonia and the pH were not limiting factors. The mortality, monitored on a daily basis, appeared after 2–51 days and was spread out over 1–20 days, depending on the increment of salinity. The higher the daily rate in salinity increase, then the shorter the time lapse before total mortality occurred. The cumulative mortality as a function of salinity fit well with simple linear regressions. The criterion of the resistance to salinity was the index MLS (median lethal salinity) defined at each daily rate as the salinity at which 50% of fish died. For S. melanotheron, the mean MLS was 123.7±3.5 g l⁻¹ whatever the daily rate in salinity. For O. niloticus, the MLS was 46.3±3.4 g l⁻¹ for daily increases in salinity ranging from 2 to 8 g l⁻¹ day⁻¹ and decreased significantly (P<0.05) above this level. The MLS-8 g l⁻¹ day⁻¹ ,which takes into account the full capacity of the fish to adapt to the increasing salinity, appeared to be a simple, optimized and efficient criterion for assessing the resistance to salinity for O. niloticus and S. melanotheron. This criterion can be a useful tool for ranking the different parental strains and hybrids of different genus and species of tilapia used in programmes of genetic selection for growth and salinity tolerance.     

广西罗非鱼链球菌病流行菌株PCR鉴定和PFGE基因型分析

为评估吉富罗非鱼(GIFT Oreochromis niloticus)抗病F₅代的选育效果,以F₀代群体和奥尼罗非鱼(Oreochromis aureus×O.niloticus)作为对照组,对F₅代42个家系的抗病性能及生长性能进行评估。对42个家系进行人工腹腔注射感染无乳链球菌 (Streptococcus agalactiae)后,分别有36个和22个家系的感染成活率比F₀代和奥尼罗非鱼高,各家系的抗病育种值为 – 0.158~0.086;生长性能测定发现,42个家系中有27个家系的绝对增长率大于F₀代;感染成活率与体质量之间的遗传相关系数为 – 0.033。综合分析各个家系的感染成活率、抗病育种值和绝对增长率,筛选出抗病力强且生长速度快的家系7个(2^#、3^#、4^#、6^#、28^#、31^#和35^#),可作为F₆代选育和苗种扩繁的亲本。结果表明,经过5个世代的针对性选育,吉富罗非鱼的抗病性能和生长性能均得到了明显改善,选育效果显著。     

The dietary protein requirements of young tilapia and an evaluation of the least cost dietary protein levels

In this study our goal was to establish the most economical dietary protein content for tilapia culture. To this end we assessed the relationship of growth, measured as percent average daily gain (% ADG), food conversion ratio (FCR) and protein efficiency ratio (PER) of young of four tilapiine species, Oreochromis mossambicus, O. niloticus, O. aureus and Tilapia zillii, for which experimental growth data were available, in relation to body weight and dietary protein content (% protein).

The relationship of % ADG to % protein in young tilapia weighing less than 1 g or 1–5 g was found to be a second order polynomial quadratic function whereas FCR and PER were linearly related to % protein. % ADG, FCR and PER were correlated better to body weight (curvilinearly) than % protein in both size groups, and multiple regressions were derived between the above parameters.

The most economical dietary protein content was evaluated from the polynomial quadratic function utilizing 95% confidence limits and also from the multiple regressions incorporating FCR and % ADG for a set of nearly isocaloric diets (gross energy) of different protein content ranging from 12% to 44%. Data derived from the foregoing approaches showed that young tilapia weighing between 1 and 5 g require 28% of the diet as protein. This dietary protein content, however, is considerably less than the protein level which supports maximum growth, namely 34%.     

Observation on the spawning of Sardina and Sardinella off the south Moroccan Atlantic coast (21–26°N)

This paper provides information on the early life stages of the Moroccan Atlantic sardine, Sardina pilchardus, and two species of Sardinella: Sardinella aurita and S. maderensis, between Cape Blanc (21°N) and Cape Boujdor (26°N), dealing with the spawning grounds, the nursery areas and the optimal spawning temperature and salinity range.

The spawning areas for Sardina are located North of Dakhla (24°30′–25°30′N) and near Cintra Bay (23°N). The larval development area is described together with seasonal and annual characteristics. The nurseries of Sardina are near Dakhla and to the south between 21 and 22°N but precise locations vary with season. The maximum occurrence of eggs was during winter, and was at temperatures from 16 to 18 °C in winter and between 18 and 18.5 °C in summer.

The main spawning area of Sardinella species is between Cape Blanc and Cintra Bay (21–23°N) with a maximum occurrence of eggs and larvae in July. For Sardinella off the southern region of Morocco, the optimal temperature interval for spawning is between 18 and 21.14 °C. No conclusions can be drawn on the relationship between spawning and water salinity.

The area north of Cintra Bay has a broad, but shallow continental shelf. This topography leads to decreasing dispersion effects and the zone constitutes a favorable area for larval retention and development.     

Effects of dietary protein level on spawning performance of Nile tilapia (Oreochromis niloticus) broodstock reared at different water salinities

Effects of dietary protein level and water salinity on spawning performance of Nile tilapia broodstock and growth of their larvae were studied. Four isocaloric (400 kcal/100 g) diets containing 25%, 30%, 35% and 40% crude protein were prepared. The diets were fed to broodfish (25.7 g) reared at three water salinities (0‰, 7‰ and 14‰) at a female/male ratio of 3:1, to satiation twice a day for 195 days. The size at first maturation increased with increasing dietary protein at all salinities. At 25% and 30% protein levels, broodstock reared at 0‰ reached their sexual maturity at bigger sizes than those reared at 7‰ and 14‰. At 0‰, spawning intervals were not significantly affected by dietary protein levels. At 7‰ and 14‰, spawning intervals significantly decreased with increasing dietary protein level. Spawning frequency and number of eggs per spawn were increased with increasing dietary protein level. The total number of spawnings per female and absolute fecundity were better in fish fed 40% protein in freshwater than at 7‰ and 14‰ salinity. The relationship of dietary protein and water salinity on egg size was significant, but showed irregular patterns. The chemical composition of broodstock muscles, eggs and fry were not significantly affected by dietary protein and water salinity, except for body water and crude protein of broodstock which were significantly affected; but showed irregular trends. At each water salinity, egg hatchability was linearly increased with increasing dietary protein level. Eggs produced from broodstock fed 25% protein at 7‰ and 14‰ needed more time for hatching and yolk-sac absorption and resulted in poorer larval weight than those reared in freshwater. Fry growth was improved with increasing protein level at all salinities. This result revealed that 40% dietary protein is required for optimum spawning performance of Nile tilapia reared at 0‰, 7‰ and 14‰ salinity. It also indicated that spawning performance and larval growth were better in freshwater than at 7‰ and 14‰.     

Comparative Nile tilapia and silver carp filtration rates of Partitioned Aquaculture System phytoplankton

Nile tilapia (Oreochromis niloticus) and silver carp (Hypophthalmichthys molitrix) held in timed-pulse feeding chambers were provided water dominated either by green algae (Scenedesmus, Ankistrodesmus and Tetraedron) or by cyanobacteria (Microcystis) to compare filtration rates (FRs). FRs were expressed as suspended particulate organic carbon (POC)/kg wet fish weight/h and as phytoplankton units filtered based on counts. Nile tilapia and silver carp filtration significantly reduced phytoplankton number of both taxonomic groups with the larger phytoplankton being filtered proportionally more than the smaller phytoplankton. Nile tilapia FR of green algae was significantly higher than silver carp; however, silver carp FR of cyanobacteria was higher than Nile tilapia. Ivlev's 90% saturation FRs (FR₉₀) in green-algal and cyanobacterial water sources were 702 and 812 mgC/kg/h for Nile tilapia and 414 and 1028 mgC/kg/h for silver carp, respectively. Silver carp were observed to reach these FR₉₀ values at lower POC concentrations than Nile tilapia with both green algae and cyanobacteria.     

The utilization of a filamentous green alga (Cladophora glomerata (L) Kutzin) as a protein source in pelleted feeds for Sarotherodon (Tilapia) niloticus fingerlings

Duplicate groups of S. niloticus fingerlings were fed each of five different diets for 8 weeks. The diets contained 30% crude protein supplied by varying proportions of fishmeal and C. glomerata meal. The five diets were formulated to supply fishmeal protein: C. glomerata meal protein ratios of 30 : 0 (diet 1), 25 : 5 (diet 2), 20 : 10 (diet 3), 15 : 5 (diet 4) and 10 : 20 (diet 5) respectively. A sixth diet (diet 6) containing 25% crude protein supplied entirely by C. glomerata meal was also fed.

Diets 1 and 2 produced significantly the highest growth rates and best protein utilization. Highest values for protein digestibility were found in diets 2 and 3 (94.5 and 93.9% respectively) although the values were not significantly different from other treatments. Weight gain and protein utilization decreased as the level of algal protein increased in diets 2 to 6. Carcass analysis revealed a decrease in lipid and protein contents with increasing levels of algae in the diet. It is postulated that algal meals are viable partial dietary protein sources for tilapia.     

Effect of probiotics, Enteroccus faecium, on tilapia (Oreochromis niloticus) growth performance and immune response

The aim of this study was to analyze the effect of a probiotic bacterium, Enterococcus faecium ZJ4 on growth performances and immune responses of tilapia (Oreochromis niloticus). The tilapias were treated with E. faecium ZJ4 at a final concentration of 1 × 10⁷ cfu ml^− 1 in aquaria water every 4 days. Six aquaria with three replicates for treated and controls were used. After 40 days, the tilapias supplemented with the probiotic showed significantly better final weight and daily weight gain (DWG) than those fed the basal diet (Control) (P < 0.05). There was no remarkable difference (P > 0.05) in the total serum protein, albumin content, globulin concentration and A/G ratio between the treated and control tilapias. The result of lysozyme activity assay was similar to these biochemical indexes. However, the complement component 3 content, myeloperoxidase (MPO) activity and the respiratory burst activity of blood phagocytes were higher (P < 0.05) in E. faecium treated tilapias (trial 1) than the controls.     

水生动物肌醇营养研究进展

采用肌醇水平分别为0 mg·kg^–1、100 mg·kg^–1、200 mg·kg^–1、400 mg·kg^–1、800 mg·kg^–1和1 600 mg·kg^–1饲料的6组等氮等能实验饲料,养殖初始体质量(74.36±5.32) g的吉富罗非鱼(GIFT Oreochromis niloticus) 12周,通过测定生长、鱼体成分以及血清指标来综合评价饲料肌醇水平对罗非鱼的影响,以期获得大规格吉富罗非鱼对饲料肌醇的需要量。结果表明,饲料中添加肌醇可显著提高罗非鱼增重率、采食量和饲料系数(P<0.05);对增重率与饲料肌醇水平作二次回归分析,得出其需要量为847 mg·kg^–1饲料;罗非鱼全鱼、肌肉和肝脏中的粗脂肪含量均随饲料肌醇添加量的增加而降低;添加肌醇对罗非鱼肌肉、肝脏粗蛋白含量均无显著影响(P>0.05)。当肌醇水平为800 mg·kg^–1和1 600 mg·kg^–1饲料时,血清谷丙转氨酶和谷草转氨酶活性均较对照组显著降低(P<0.05);随饲料中肌醇含量的升高,总胆固醇和甘油三酯含量呈现先降低后升高的趋势,而血糖呈现先升高后降低的趋势。综上所述,大规格吉富罗非鱼饲料中肌醇的需要量为847 mg·kg^–1饲料,且饲料中添加肌醇有利于降低组织脂肪蓄积,并可改善肝功能。     

半滑舌鳎(Cynoglossus semilaevis)黑色素富集激素基因的克隆和表达

采用同源克隆技术获得了佛罗里达红罗非鱼(Oreochromis sp.)前原黑色素浓集激素1 (prepro-melanin concentrating hormone 1,pmch1)基因cDNA全长序列,将其命名为flpmch1;并通过实时荧光定量PCR (qRT-PCR)分析天生黑斑佛罗里达红罗非鱼组织表达谱和低温胁迫后其各组织表达差异。结果表明,flpmch1 cDNA序列全长629 bp,开放阅读框411 bp,编码136个氨基酸,存在12个潜在磷酸化位点和1个长度为199 bp的CpG岛。Flpmch1与24个物种Pmch1氨基酸序列比对结果表明,Flpmch1与尼罗罗非鱼(O.niloticus)、奥利亚罗非鱼(O.aureus)、莫桑比克罗非鱼(O.mossambicus)、布氏新亮丽鲷(Neolamprologus brichardi)的Pmch1相似性最高。系统进化树分析显示硬骨鱼纲Pmch聚成一个大支,Flpmch1与尼罗罗非鱼Pmch1进化地位最接近。组织表达谱分析显示,flpmch1 mRNA在多个组织中均有表达,其中在脑中表达量最高,且粉白色皮肤中的表达量显著高于黑色皮肤(P<0.01)。低温胁迫后flpmch1 mRNA各组织表达量较对照组均呈下调表达。推断flpmch1可能参与了佛罗里达红罗非鱼天生黑斑的形成和过冬黑化过程。     

Dietary impact of algal and artificial diets, fed at different feeding rations, on the growth and fatty acid composition of Tapes philippinarum (L.) spat

The effect of lipid supplementation and algal ration on growth and fatty acid composition of juvenile clams, Tapes philippinarum was investigated. A diet of Tetraselmis suecica was supplemented with a lipid emulsion and fed at a daily ration of 0.5, 1.0 and 1.5% WW DW⁻¹ day⁻¹. A mixed algal diet of Isochrysis galbana (clone T-Iso) and T. suecica (1:1 on DW basis) and starvation functioned as positive and negative control treatments, respectively. T. suecica, which contains eicosapentaenoic acid (20:5n−3) but no docosahexaenoic acid (22:6n−3), was supplemented with 50% (% of algal dry weight) of a 22:6n−3-rich emulsion. Higher growth rates of T. philippinarum were associated with higher T. suecica feeding rations. Lipid supplementation resulted in significantly better growth rates of T. philippinarum at a T. suecica ration of 0.5, 1.0 and 1.5% WW DW⁻¹ day⁻¹. The growth promoting effect of lipid supplementation was significantly reduced during 4 consecutive weeks or by increasing the algal feeding ration. Increasing T. suecica rations were associated with a decrease in the % composition of 22:6n−3 while the 20:5n−3% composition was hardly affected in the total lipids of T. philippinarum. On the contrary, the absolute concentration of 22:6n−3 (μg per gram dry weight, μg g DW⁻¹) in the total lipids of T. philippinarum was similar at all T. suecica rations while the 20:5n−3 concentration significantly increased. The major effect of lipid supplementation on the fatty acid composition of the clams, was a significant increase of the 22:6n−3% composition in the total lipids, polar lipids and triglycerides. In contrast with 20:5n−3, the effect of lipid supplementation on the absolute 22:6n−3 content (μg g DW⁻¹) was more pronounced at a higher T. suecica ration. After a starvation period of 4 weeks, the concentration (μg g DW⁻¹) of all fatty acids was significantly reduced, except the n−7 monoenoic fatty acids (MUFA) and the non-methylene interrupted dienoic fatty acids (NMID). The concentration (μg g DW⁻¹) of the NMID in starved clams and clams fed different diets was similar to the concentration in the initial sample. This indicated that growing spat was actively synthesising NMID and they were selectively retained in unfed animals. The results indicated that either dietary lipid or 22:6n−3 was a growth-limiting factor when T. suecica was fed at a ration of 0.5, 1 and 1.5% WW DW⁻¹ day⁻¹. A selective retention in starved clams and a preferential accumulation in fed clams was observed for 22:6n−3 but not for 20:5n−3 which indicated a greater importance of 22:6n−3 compared to 20:5n−3.     

Transgenic Nile tilapia (Oreochromis niloticus) over-expressing growth hormone show reduced ammonia excretion

The growth of aquaculture has negatively affected the environment due to the high levels of nitrogen excreted by farmed fish. Here we propose that modifying the nitrogen metabolism of the fish themselves using transgenic technology might solve the pollution problem. Growth hormone (GH) is known to increase protein retention and absorption, and is thought to reduce ammonia excretion. Thus, we produced transgenic Nile tilapia (Oreochromis niloticus) that over-expressed the GH gene throughout their bodies. Our findings showed that the food-conversion efficiency of the transgenic fish was 35% higher than that of their non-transgenic siblings. The rearing period required for the transgenic fish to reach a body weight of 20 g was about 75% of that required for non-transgenic fish that were fed the same type and quantity of food. The total amount of ammonium-nitrogen excreted by the transgenic fish was about 69% of that excreted by the wild-type fish over their lifetime. These results suggest that our transgenic approach has the potential to reduce the amount of nitrogen pollution caused by farmed fish. This strategy is a promising option for making aquaculture more ‘eco-friendly’.     

Preliminary tests of an aerated tank system for tilapia culture

A simple tank system, using artificial aeration to supply oxygen and incoming water to dilute metabolic by-products and other wastes, was tested. Blue tilapia, Oreochromis aureus, were grown under a wide range of loads, 7–31 kg liter⁻¹ min⁻¹, and densities, 19–77 kg fish m⁻³ culture unit. Average yield ranged from 6–10 kg m⁻³ culture unit per month. Approximately 14–20 m³ of water was used for each kilogram of fish produced.     

The effects of ozone and probiotics on the survival of black tiger shrimp (Penaeus monodon)

Total ozone production (TOP) from an ozonator, residual ozone concentration (ROC) in water, and the effects of ozone with or without probiotic supplemented feeds on bacterial growth, and shrimp (Penaeus monodon) survival were investigated. Minimal effective ROC to inhibit 3 log units of Vibrio harveyi D331 for 6 h and 2 log units of Bacillus S11 for 9 h was 0.38 g O₃/l of ROC from 5-min ozonation. Shrimp postlarvae exposed to 0.34–0.50 mg O₃/l ROC (8-h ozonation) caused loss of balance, immobility and destruction of gill lamellar epithelium. In vivo treatment of juvenile P. monodon reared on probiotic feed for 1 month revealed that 0.35 mg O₃/l ROC (30-min ozonation) effectively inhibited 3 log units of V. harveyi D331 for 24 h. At this ROC dosage, there was no effect either on shrimp or on intestinal probiotics (Bacillus S11). Shrimp survival from probiotic treatment, coupling with ozonation, increased significantly (P≤0.05) compared with controls.     

Passive transfer of protective immunity against ichthyophthiriasis from vaccinated mother to fry in tilapias, Oreochromis aureus

Tilapias, Oreochromis aureus, were vaccinated twice with live tomites of Ichthyophthirius multifiliis in physiological saline 1 month before spawning (test group). A control group was similarly treated but with physiological saline only. Fertilized eggs were collected from the mouths of both groups and incubated until the young reached the free-swimming stage. Free-swimming fry were also collected immediately after they were expelled from the mother's mouth in other fish of the two groups. The fry were exposed to infective tomites of I. multifiliis and thereafter returned to their holding tanks. All fry obtained from the controls without mouth-brooding died after tomite exposure. However, fry with mouth-brooding before tomite exposure showed 37.3% survival. On the other hand, fry obtained from vaccinated broodstock without mouth-brooding exhibited 78.4% survival which increased to 95.3% with mouth-brooding. The protective immunity is correlated with the titres of anti-I. multifiliis antibodies in the soluble extracts of fry tissues and the mother's plasma. The results clearly indicate that protective immunity against ichthyophthiriasis “ich” in tilapia fry can not only be derived directly from the mother via eggs but also be acquired indirectly from the mouth cavity during the brooding period. The results thus suggest that proper vaccination of mothers before spawning would effectively protect fry against a protozoan disease.     

Dietary administration of an LHRH analogue induces spawning of spotted seatrout (Cynoscion nebulosus)

The effects of a single oral administration of des-Gly¹⁰-[D-Ala⁶]-luteinizing hormone-releasing hormone (1–9)-ethylamide (LHRHa) in the diet (0.2–2.5 mg/kg body weight) on ovulation and spawning of spotted seatrout (Cynoscion nebulosus) were examined. Oral administration of 1.0–2.5 mg LHRHa/kg to females in four separate experiments resulted in successful spawning 32–38 h later with mean fertilization and hatching success rates of 93.3% and 74.6%, respectively. A lower dose of LHRHa (0.2 mg/kg) was ineffective in two subsequent experiments. The data suggest that oral administration of 1 mg LHRHa/kg is a reliable method of inducing ovulation in spotted seatrout, resulting in predictable spawning within 38 h of feeding. This noninvasive treatment may be particularly useful for induced spawning of teleosts which are susceptible to handling stress.     

Effect of water temperature on the immune response of white shrimp Litopenaeus vannamei to Vibrio alginolyticus

White shrimp Litopenaeus vannamei held in 25‰ seawater at 27 °C or 28 °C were injected with TSB-grown Vibrio alginolyticus at 1 × 10⁴ colony-forming units (cfu) shrimp^− 1 or 1 × 10⁵ cfu shrimp^− 1, and then cultivated onward at water temperatures varying from 20 to 34 °C. Over 24–144 h, mortality of V. alginolyticus-injected shrimp held at 34 °C or 32 °C was significantly higher than that of shrimp held at lower temperatures. In a separate experiment, shrimp held in 25‰ seawater at 28 °C and then cultured onward at 20 to 32 °C were examined for immune parameters at 24–96 h. THC, phenoloxidase activity, respiratory burst, and SOD activity decreased significantly at 24 h after transfer to 32 °C. Shrimp held in 25‰ seawater at 27 °C and then cultured onward at 20 to 34 °C showed a significant reduction in phagocytic activity and clearance efficiency for V. alginolyticus at 24 h after transfer to 34 °C. It was concluded that transfer of shrimp from 27 or 28 °C to higher temperatures (32 and 34 °C) reduced their immune capability and decreased resistance to V. alginolyticus infection.     

Production of a high percentage of male offspring in growth-enhanced transgenic tilapia using Oreochromis aureus ZZ selected pseudofemales

Gene transfer has offered a new tool for the development of improved fish strains for aquaculture. Monosex fish populations could minimize the impact of genetically modified organisms in the environment. In Oreochromis aureus, the use of pseudofemale spawners (sex-reversed male with a female phenotype) is an alternative technique for producing genetically male tilapia offspring. O. aureus fry were treated with 17β-estradiol at 100 mg/kg of food for 45 days. We obtained 77.1% females and 45.9% in the control group. Females randomly taken from the treated group were crossed with normal males. Fry from pseudofemales producing more than 90% male progenies were submitted to 17 β-estradiol treatment to obtain F₂ pseudofemales. The results of the sex-reversal were low and variable ranging between 66.0 and 84.3% females. F₂ pseudofemales were crossed with transgenic males from the F70 line (O. aureus × O. urolepis hornorum) and non-transgenic (O. aureus) males. The sex ratio of progeny of F₂ pseudofemale deviated significantly (P < 0.01) in favor of males in the crosses with transgenic (90.2%) and non-transgenic (89.3%) males compared to the results observed with normal females (51.0 and 52.3%, respectively). The mean fry production with pseudofemales (per m²/day) was similar to the normal females in the crosses with transgenic and non-transgenic males. To our knowledge this is the first report on the production of a near monosex population in genetically modified fish.     

Hatchery production of diploid and triploid clams, Tapes dorsatus (Lamarck 1818): a potential new species for aquaculture

To assess the potential for hatchery production of the venerid clam, Tapes dorsatus, diploid and triploid clams were produced and ongrown until the spat reached 5–8 mm in size. Triploidy percentages at metamorphosis that ranged from 56 to 85% were induced using a 15-min exposure to 1 mg/l cytochalasin B. No differences were observed in the growth rate of sibling diploid and triploid T. dorsatus larvae. Survival of diploid clams to pediveliger stage was however higher. Post-metamorphic growth and survival of diploid and triploid clams remained the same until the clams were removed from the hatchery to outdoor nurseries. Throughout the larval and early spat phases of production, T. dorsatus growth was similar to that reported for the Manila clam, Tapes philippinarum. Based upon its ease of culture, rapid growth and marketability, T. dorsatus are thought to have considerable aquaculture potential.     

A study of appetite, digestion and growth in juvenile green turtle (Chelonia mydas L.) fed on artificial diets

One-year-old immature green turtles (500–900 g) were kept at 25°C in recirculating seawater. When deprived of food for 108 h and then offered floating trout pellets, they consumed a satiation meal (S g) described by the equation S = 0.025 W^0.96 where W is wet body weight in g. When food was offered at different deprivation times after satiation, maximum meals were taken after 60–72 h and corresponded to 2.5% body weight. Longer deprivation (108 h) led to reduced intake of 1.9–2% body weight. Gastric emptying was measured using X-radiography or chromic oxide to label the faeces. Complete stomach emptying required 110 ± 11 h S.D. and maximum appetite occurred when the stomach was estimated to be 80–95% empty. The average time for a meal to be voided from the alimentary tract was 176 ± 16 h S.D. Diets containing 40–50% protein and 4.2–5 kcal/gram were assimilated with apparent efficiencies of 76 ± 6% S.D. and 86 ± 6% for energy and protein nitrogen, respectively. A preliminary estimate for an energy budget for these animals based on a 23 day growth period accounted for 90% of the daily food intake as:

100 I39M + 32E

where known daily intake (I calories) is compared with measures or estimates of metabolic rate (M), growth (G) and excreted wastes (E). Although this balance is similar to that of herbivorous fish, the turtle has slower feeding and digestion rates than its teleost counterparts.