Carbonic anhydrase and Na/K-ATPase activities during the molt cycle of low salinity-reared white shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

Changes in hemolymph osmolality, ion concentrations, and enzymatic activities of carbonic anhydrase (CA) in the gills and epidermal tissue, and Na/K-ATPase in the gills during the molt cycle were investigated in the white shrimp Litopenaeus vannamei. Hemolymph osmolality was high in the intermolt and early premolt stages, but started to decrease prior to ecdysis through to postmolt stages A and B. Changes in Na⁺ and Cl⁻ ion concentrations paralleled those in hemolymph osmolality. CA activity levels in the anterior and posterior gills were low at intermolt stage C₀ and premolt stage D₀, and maximum at premolt stage D₃. In the epidermal tissue, activity was relatively high at intermolt stage C₀ and premolt stage D₀, but fluctuated towards premolt stage D₃ and postmolt stage A. On the other hand, Na/K-ATPase activity in the gills decreased between intermolt stage C₀ and premolt stage D₂, but increased at premolt stage D₃ and postmolt stage A. The changes in patterns of CA activity during the molt cycle suggest that CA may be involved in supplying counter-ions for Na⁺ and Cl⁻ uptake during molting. Branchial Na/K-ATPase appears to be involved in producing local osmotic gradients in order to support water influx across the epithelium.     

Nutritional evaluation of waste date fruit as partial substitute for soybean meal in practical diets of juvenile Nile tilapia, Oreochromis niloticus L.

The potential of waste date meal (WDM; low-quality date palm, Phoenix dactylifera L.) as a carbohydrate source in formulated diets for Nile tilapia was evaluated. Four isocaloric-practical diets (15.7 kJ g⁻¹) were formulated incorporating WDM at 0, 100, 200 and 300 g kg⁻¹ levels as partial substitutes for soybean meal (SBM). These were designated D₀ [284 g crude protein (CP) and 383 g carbohydrate (CHO) kg⁻¹ diet], D₁ (279 g CP and 446 g CHO kg⁻¹ diet), D₂ (207 g CP and 495 g CHO kg⁻¹ diet) and D₃ (175 g CP and 578 g CHO kg⁻¹ diet). Each diet was fed to three replicate groups of 30 fish [20.20 ± 0.09 g (±SE)] for 75 days. No feed-related mortality was observed during the entire experimental period. Final body weight (FBW) and specific growth rate (SGR) in the different treatments were statistically not significantly different ( P  > 0.05). Protein efficiency rate (PER) was lowest in diet D₀ and increased with decrease of SBM content (D₁–D₃). A significant increase in whole body lipid content was recorded in fish fed diets D₂ and D₃. Results showed that WDM could be a substitute for SBM up to 300 g kg⁻¹ in practical Nile tilapia diets without compromising growth.     

Effects of hydrogen peroxide on clearance of formalin-killed Flavobacterium branchiophilum from the gills of rainbow trout, Oncorhynchus mykiss (Walbaum)

The effect of hydrogen peroxide (H₂O₂) treatment on the clearance of formalin-killed Flavobacterium branchiophilum from the gills of rainbow trout was examined. In untreated control fish, clearance was characterized by a rapid initial phase, with 50% reduction in bacterial antigen in the first 12 h after exposure. The bacteria then cleared more gradually, with total clearance being achieved by 40 h. Treatment of fish with 100 p.p.m. H₂O₂ did not influence bacterial clearance compared to untreated controls. Exposure for 1 h to H₂O₂ concentrations ≤ 100 p.p.m. caused no detectable clinical signs or evidence of ultrastructural damage to the respiratory epithelium. However, levels in excess of this caused significant gill damage.     

Three different levels of dietary vitamin D3 fed to first-feeding fry of Atlantic salmon (Salmo salar L.): effect on growth, mortality, calcium content and bone formation

Atlantic salmon ( Salmo salar L.) fry were reared on a fishmeal-based diet with three different levels of vitamin D₃ (0.2, 5 and 57 mg vitamin D₃ kg^–1 feed, ww) from first-feeding for 14 weeks. No significant differences were recorded in weight, length, specific growth rate, mortality, or kidney calcium concentration between the different dietary groups. No skeletal malformations or histopathological changes were recorded in any of the dietary groups. These results suggest Atlantic salmon fry to be highly tolerant of megadoses of vitamin D₃ over a limited period of time.     

Sensitivity of juvenile lobsters, Homarus americanus L., to different concentrations of malachite green during various states of the intermoult cycle

Abstract. Survival of the early stages (egg, larva and juvenile) of the American lobster, Homarus americanus L., reared under controlled conditions has often been threatened by a variety of fungi and microbial epibionts. A preliminary study was conducted for the purpose of defining toxicity levels of malachite green for 5th stage juveniles during each of the following intermoult (or fused intermoult) states: A-B, C₄, D₀ and D₂-D₃ Irrespective of exposure time or concentration, all the lobsters treated during intermoult state A-B died. This contrasts with seemingly good survival of lobsters in intermoult states C₄, D₀ and D₂- D₃ treated at concentrations of 900 mg/1 malachite green for exposure periods of 10 min. The findings suggest that an effective treatment combination for lobsters immersed in malachite green may be one which is employed during intermoult state C₄, for about 6min and at concentrations of about 700–800 mg/l.     

In vitro methyl farnesoate secretion by mandibular organs isolated from different molt and reproductive stages of the crab Oziotelphusa senex senex

ABSTRACT:   Considerable evidences indicate that mandibular organ (MO) of crustaceans secrete terpenoid hormone (methyl farnesoate [MF]), which regulates molting and reproduction. In the freshwater south Indian rice field crab Oziotelphusa senex senex , MO isolated during premolt and vitellogenesis, secreted greater amounts of MF, which indicates that the regulation of the crustacean molt and reproduction is complex and also involves MF, besides steroid (ecdysteroid) and peptide (sinus gland peptide) hormones.     

Effect of Dietary Supplementation of Vitamins A, D3, E, and C on Yearling Rice Field Eel, Monopterus albus: Serum Indices, Gonad Development, and Metabolism of Calcium and Phosphorus

Abstract.— The objective of this study was to determine the effect of dietary vitamins A, D₃, E, and C on the gonad development, lipid peroxidation, and immune response of yearling rice field eel, Monopterus albus . A 6-wk feeding trial was designed according to an L₁₆(4⁵) orthogonal design, in which four vitamins, each at four supplementation levels, were arranged. Sixteen diets were mixed with the different vitamin levels and randomly assigned to 16 groups of fish. Increasing dietary vitamin E supplementation level significantly ( P  ≤ 0.05) increased the gonadosomatic index and lowered the serum content of malondialdehyde of rice field eel. Increasing dietary vitamin A and C levels also showed similar effect, but the differences were not statistically significant. Serum immunoglobulin M content increased significantly ( P  ≤ 0.01) as dietary vitamin C supplementation levels increased. The concentrations of calcium in bones showed significant ( P  ≤ 0.05) trend with vitamin D₃ and A supplementation levels, but the bone phosphorus content was not affected by the dietary vitamin levels.     

蜕壳周期内中华绒螯蟹钙含量、组织结构及相关基因表达变化

运用生化分析、组织切片和分子生物学等技术,测定蜕壳期(蜕下旧壳后0.5 h内)和蜕壳后期(蜕下旧壳0.5 h后至新壳未完全硬化之前)平均体质量(5.63±2.35)g中华绒螯蟹肝胰腺和鳃中的钙含量,观察肝胰腺和鳃组织结构变化,用荧光定量PCR技术分析肝胰腺和鳃中的钙网蛋白(CRT)基因的表达情况.试验结果表明,蜕壳后期...     

Tissue distribution of vitamin D3 in Atlantic salmon Salmo salar: effect of dietary level

Atlantic salmon Salmo salar were given three dietary doses of vitamin D₃ for a period of 11 weeks: diet groups I, II and III received a 0.04, 2.21 and 28.68 μg g⁻¹ diet of vitamin D₃, respectively. The tissue distribution of vitamin D₃ was investigated, and between diet groups II and III, the level of vitamin D₃ found in the tissues was increased by a factor similar to the increased level in the feed. No effects were observed on weight gain, survival, plasma level of calcium, red blood cell count or haematocrit, relative to the dietary levels of vitamin D₃. The results showed that while the plasma concentration of 25(OH)D₃ increased between diet groups II and III, the concentration of 1,25(OH)₂D₃ decreased.     

Histopathology of rainbow trout, Salmo gairdneri Richardson, exposed to diurnally fluctuating un-ionized ammonia levels in static-water ponds

Abstract. Rainbow trout, Salmo gairdneri Richardson, were reared intensively in nine aerated static-water ponds. Dissolved oxygen concentrations were maintained above 5 mg/l and un-ionized ammonia (NH₃) levels were monitored in all ponds. Fish were removed periodically and gill, liver and trunk kidney samples were removed for histological examination. Average NH₃ concentrations did not exceed a previously reported chronically toxic level, but average daily maximum concentrations exceeded this level in all but two ponds. Gills exhibited lesions characteristic of those attributed to ammonia exposure in all ponds and fish with the most damaged gills had livers demonstrating reduced glycogen vacuolation. No histological changes were noted in kidney tissue. Because gills were damaged even though average NH₃ levels were quite low, it is recommended that research in static water carefully consider the cyclical nature of NH₃ concentrations.     

Further development of bithionol therapy as a treatment for amoebic gill disease in Atlantic salmon, Salmo salar L.

This study examined the efficacy of bithionol as a prophylactic or therapeutic oral treatment for Atlantic salmon (AS), Salmo salar , affected by amoebic gill disease (AGD). Furthermore, it explored the interaction of bithionol oral therapy with the current standard treatment (a freshwater bath for at least 3 h). The efficacy of three medicated feeds was determined in the trial by feeding AGD-affected AS at 1% body weight (BW) day⁻¹ either oil coated commercial feed (control) or prophylactic and therapeutic bithionol at 25 mg kg⁻¹ feed. Feeding commenced 2 weeks prior to exposure to Neoparamoeba spp. at 300 cells L⁻¹ and continued for 49 days post-exposure (PE). Bithionol when fed as a 2-week prophylactic or therapeutic treatment at 25 mg kg⁻¹ feed delayed the onset of AGD pathology and reduced the percentage of gill filaments with lesions. Administration of a 3-h freshwater bath at 28 days PE significantly reduced amoeba numbers to a similar level across all treatments; in contrast, gross gill score and percent lesioned filaments were reduced to different extents, the control having a significantly higher score than both bithionol treatments. Following the freshwater bath, clinical signs of AGD increased at a similar level across all treatments, albeit controls were significantly higher than the bithionol treatments immediately following freshwater treatment. This study demonstrated that bithionol at 25 mg kg⁻¹ feed, when fed as a 2-week prophylactic or a therapeutic treatment, delayed and reduced the intensity of AGD pathology and warrants further investigation as a treatment for AGD-affected AS.     

Liver oil of pharaoh cuttlefish Sepia pharaonis Ehrenberg, 1831 as a lipid source in the feed of giant freshwater prawn, Macrobrachium rosenbergii (De Man 1879)

The efficacy of pharaoh cuttlefish ( Sepia pharaonis Ehrenberg, 1831) liver lipid in the feed for juveniles of giant freshwater prawn Macrobrachium rosenbergii (De Man 1879) was tested by feeding five experimental diets prepared using clam meat, groundnut oil cake, wheat bran, tapioca powder, vitamin–mineral mixture and cellulose powder as the major ingredients. The test diets T₁, T₂, T₃, T₄ and T₅, containing five levels (1, 2, 3, 4 and 5%, respectively) of lipid, extracted from the liver of pharaoh cuttlefish, were fed to 30-day-old prawn juveniles for 60 days, in triplicates and compared against a control. Analysis of variance of the growth parameters showed that the juveniles fed with T₂, containing 2% cuttlefish liver lipid with a total lipid content of 9.85% and lipid–carbohydrate ratio of 1 : 3.8, showed significantly better growth ( P  < 0.05) with the highest weight gain of 118%, greatest food conversion and protein utilization efficiency, and the highest body protein content, although the survival rates remained unaffected among treatments. Growth of the juveniles, however, declined at lipid levels >2%. Addition of cuttlefish liver lipid led to an increase of ω-3 and ω-6 fatty acids in T₂, the proportion of ω-6 being higher than the former. The crude lipid content of the body of test animals also increased with the increase in dietary lipid from 1.51% in T₁ to 3.27% in T₅. The present study indicates that cuttlefish liver lipid can be successfully used as a substitute for conventional lipid sources in rearing juveniles of giant freshwater prawn, an eco-friendly practice in recycling cuttlefish waste – a discarded fishery by-product.     

In vitro hydroxylation of vitamin D3 and 25-hydroxy vitamin D3 in tissues of Atlantic salmon Salmo salar, Atlantic mackerel Scomber scombrus, Atlantic halibut Hippoglossus hippoglossus and Atlantic cod Gadus morhua

The in vitro metabolism of ¹⁴CD₃ and ³H25OHD₃ was investigated in different tissues from Atlantic salmon Salmo salar , Atlantic mackerel Scomber scombrus , Atlantic halibut Hippoglossus hippoglossus and Atlantic cod Gadus morhua . The tissues analysed were liver, kidney, head kidney, gills, spleen and intestine. The metabolites were extracted in methanol–chloroform and separated by normal-phase high-pressure liquid chromatography (HPLC) followed by scintillation counting. Identification of the metabolites was by comigration with standards on normal and reversed-phase HPLC systems and by protein-binding assays. All tissues from all species analysed produced hydroxylated derivatives identified as 25OHD₃, 24,25(OH)₂D₃ and 1,25(OH)₂D₃. In addition, some unidentified derivatives were recorded, one probably being 25,26(OH)₂D₃. Organs producing great amounts of one metabolite also produced considerable amounts of the other possible derivatives, suggesting a lower degree of specificity in fish organs than in human organs. The predominating metabolite was 24,25(OH)₂D₃ in all organs from salmon and mackerel during incubation with ¹⁴CD₃ and within most organs from all species during ³H25OHD₃ incubation. The latter observation probably results from the need for decreasing rather than increasing the calcium absorption in these species, which live at least some periods of life in a marine environment.     

Effects of organic acids on growth and phosphorus utilization in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:   A feeding experiment was conducted to investigate the effects of various organic acids and supplements on the growth, phosphorus (P) utilization, and environmental loading of nitrogen (N) and P in red sea bream Pagrus major. Seven diets designated as PA (0.5% inorganic P as Ca[H₂PO₄]₂), PO (no inorganic P), CA (1% citric acid), MA (1% malic acid), LA (1% lactic acid), MHA (1% methionine hydroxy analog) and LTE (1% liquid trace elements) were formulated. Duplicate groups of 25 fish were fed four times daily to near satiation for 75 days. Fish fed CA and LTE diets showed significantly better weight gain and feed conversion ratio compared to fish fed diet PO. Absorption of P was significantly improved in all organic acid-supplemented groups compared to the PA and PO groups. The P retention in CA and LTE groups were significantly higher compared to PA and PO diets. The P excretion was also significantly decreased in all organic acid-supplemented groups. Thus, use of organic acids, in particular CA and LTE, in red sea bream diets can reduce the use of inorganic phosphorus and contribute to the development of much-desired eco-friendly diets.     

Elevation of lipid peroxide level and production of hydroxy lipids in cultured Hepa-T1 cells by oxidative stressors

ABSTRACT:   In order to investigate the relationships between levels of lipid peroxides (LPO), hydroxy lipids (L-OH) and of antioxidative enzymes in live fish, cultured cells from tilapia liver (Hepa-T1) were treated with oxidative stressors, 2,2'-azobios(2-amidino-propane) dihydrochloride (AAPH) and H₂O₂. By treating with 5 mM AAPH, LPO and L-OH of the cells significantly increased time-dependently. In particular, L-OH increased from 7.6 ± 0.7 to 23.6 ± 1.8 nmol/mg protein after 2 h of treatment, and synchronously the glutathione peroxidase (GPx) activity of the cells increased from 259 ± 106 to 1970 ± 135 mU/mg protein. In the case of H₂O₂, however, no elevation of L-OH was observed. In this case, catalase (CAT) and superoxide dismutase (SOD) activities increased time-dependently. Especially, after 2 h exposure, the CAT activity increased remarkably from 217 ± 21 to 2510 ± 120 mU/mg protein, though the GPx activity did not change significantly. These results suggest that the elevation of the L-OH level is closely associated with the elevation of GPx activity, but not with that of CAT or SOD. However, when vitamin E-enriched Hepa-T1 cells were treated with AAPH and H₂O₂, the production of LPO and L-OH was suppressed significantly. The findings suggest that vitamin E also plays an important role in the levels of the oxidation products in vivo .     

Blood electrolyte and prolactin regulation in rainbow trout, Oncorhynchus mykiss (Walbaum), exposed to nitrite

Abstract. Rainbow trout, Oncorhynchus mykiss (Walbaum), exposed to environmental nitrite for 3h showed significantly reduced chloride concentrations from 120·9 ± 0·5 to 113·8 ± 2-4 mmol/1, the result of nitrite uptake by the gills with a compensatory loss of blood chloride. This change could be prevented by injection of 10 mg/kg sulpiride, which stimulates prolactin secretion through antagonism of dopamine D₂ receptors, thus increasing circulating prolactin levels. Prolactin is known to decrease membrane permeability and the increased levels of the hormone probably account for the relatively unchanged (compared to controls) plasma chloride and potassium levels in nitrite-exposed, sulpiride-treated fish.     

14C-glucose injection in Atlantic cod, Gadus morhua, metabolic responses and excretion via the gill membrane

In the present study cod were separately placed in fish metabolic chambers. Ten individuals were injected with a dose of 1 μCi ¹⁴C-glucose, in addition to 1 g pure glucose per kg live weight, and 10 individuals were kept under the same conditions and sham injected with 9 g L⁻¹ (0.9%) sterile saline solution.
Of the injected ¹⁴C-glucose, 3% was recovered in plasma, 2.9% in liver, 1.7% in red muscle, 18.2% in white muscle, 0.2% in heart, 1.9% in gills and 12.1% in gill-surrounding water. Of the 12.1% in gill-surrounding water, 9.1% was found to be in the form of CO₂, leaving 3% to pure glucose and lactate. These results indicate a relatively high production of CO₂ from glucose showing a utilization for energy, and a possible route of excretion of excess glucose via the gill membrane.
Activity of ¹⁴C-glucose on a concentration basis showed in descending order the following organs to be metabolically active in glucose utilization: gills > plasma (as a transporter) > heart > red muscle > liver > white muscle.
Of the total amount of ¹⁴C-glucose injected, only 0.3% was recovered in the liver lipid fraction, and mainly as triacylglyceroles; minor ¹⁴C-activity was found in mono- and diacylglyceroles and free fatty acids. These results show very low activity of de novo syntheses of lipid from injected glucose in the cod.     

Biotin in practical fish-meal based diet for Atlantic salmon Salmo salar L. fry

Groups of Atlantic salmon fry (0.19 g initial weight) were fed a fish-meal based starter diet with different supplements of biotin (0, 0.5, 1.0 and 1.5 mg biotin kg^–1) in triplicate tanks for 18 weeks. The basal diet contained 0.3 mg total biotin kg^–1. The experimental design included a negative control diet made by replacing 10% of the fish-meal with spray-dried raw hen's egg white.
Throughout the experimental period the fish grew to about 5 g and there were no significant differences in growth and mortality among the groups of fish fed the fish-meal diets. At the end of the experiment there were no significant differences in biotin level in the liver, while whole-body biotin concentrations correlated significantly with the dietary biotin concentrations. No significant increase in pyruvate carboxylase (PC) activity was found in livers from fish given different dietary levels of biotin. The control diet with egg white resulted in severe growth reduction and increased mortality compared with the other dietary groups. The concentrations of biotin in liver and whole body were decreased in fish fed egg white. Increased levels of glycogen and reduced PC activity in the liver were observed in this group after 18 weeks.
Histology of the gills showed no differences in appearance when fish were fed fish-meal based diets while the addition of egg white resulted in hypertrophy and hyperplasia of the gill tissue and extensive fusions of the secondary gill lamellae.
The results show that there is no need for supplemental biotin in practical fish-meal based diets for Atlantic salmon fry to achieve optimal growth, survival and maximal liver PC activity.     

Metabolic costs of changing the cation–anion difference in the diet of juvenile African catfish Clarias gariepinus (Burchell)

The influence of dietary cation–anion difference (CAD, Na⁺ + K⁺– Cl^–, mEq kg^–1) on energy metabolism and nitrogen losses in juvenile African catfish Clarias gariepinus (Burchell) was examined in fish exposed to different dietary CAD levels (–146, 116, 497, 713 and 828 mEq kg^–1 diet). The experiment was conducted in open circuit balance respiration chambers over a 3-week period. Five 24-h monitoring periods were carried out at 3-day intervals during the experimental period with O₂ consumption, ammonia and nitrate + nitrite (NO_x) and CO₂ production being measured at 5-min intervals for each chamber. The negative dietary CAD (–146 mEq kg^–1) resulted in the highest energy expenditures (83 kJ kg^–0.8· d^–1). With increasing dietary CAD levels, heat loss gradually decreased to minimum values of 56 kJ kg^–0.8 day^–1 at a dietary CAD level of 713 mEq kg^–1. Consequently, metabolizable energy utilization efficiency (MEU, percentage of retained energy over metabolizable energy) quadratically ( P  < 0.05) increased and reached a maximum at a dietary CAD of 713 mEq kg^–1.