Effects of cadmium on the kinetics of calcium uptake in developing tilapia larvae, Oreochromis mossambicus

The toxic effects of Cd²⁺ on Ca²⁺ influx kinetics in developing tilapia (Oreochromis mossambicus) larvae were evaluated. Addition of 20 µg l^-1 of Cd²⁺ to the environment of 0 and 3 day-old larvae competitively inhibited the Ca²⁺ uptake within 4h resulting in a great increase in K_m values for Ca²⁺ influx (19.3 and 17.4 fold, respectively) as compared with their respective controls. Consequently, the actual Ca²⁺ influx of larvae in solutions of 0.2 mM Ca²⁺ are suppressed by 32–45%. Also, 3 day-old larvae were more sensitive to internally accumulated Cd²⁺ than 0 day-old larvae. Although the Ca²⁺ influx in 0 and 3 day-old larvae may be restored to the levels of their respective controls with 24h of being transferred to a 20 µg l^-1 Cd²⁺ solution, total body Ca²⁺ content was significantly reduced in 3 day-old larvae. Increased Ca²⁺ uptake efficiency ensures sufficient Ca²⁺ for normal growth. However, rapid increase in Ca²⁺ influx after hatching also leads to higher Cd²⁺ uptake. Exposure to Cd²⁺ will lead to a drop in body Ca²⁺ content resulting in retardation of larval growth. Therefore, we conclude that if Ca²⁺ uptake is interfered with at this critical stage of development, larvae will not be able to maintain normal levels of body Ca²⁺ and will show signs of Cd²⁺ poisoning.     

Calcium balance in embryos and larvae of the freshwater-adapted teleost,Oreochromis mossambicus

Changes in Ca²⁺ content and flux, and the development of skin chloride cells in embryos and larvae of tilapia, Oreochromis mossambicus, were studied. Tilapia embryos hatched within 96h at an ambient temperature of 26–28°C. Total body Ca²⁺ content was maintained at a constant level, about 4–8 nmol per individual, during embryonic development. However, a rapid increase in body Ca²⁺ level was observed after hatching, 12.8 to 575.3 nmol per individual from day 1 to day 10 after hatching. A significant influx and efflux of Ca²⁺ occurred during development, with the average influx rate for Ca²⁺ increasing from 5.9 pmol mg⁻¹ h⁻¹ at 48h postfertilization to 47.8 pmol mg⁻¹ h⁻¹ at 1 day posthatching. The skin was proposed as the main site for Ca²⁺ influx before the development of gills, and the increased Ca²⁺ influx may be ascribed to gradual differentiation of skin surface and chloride cells during embryonic development. Ca²⁺ efflux was 16–56 pmol mg⁻¹ h⁻¹ in 1-day-old larvae. The resulting net influx of Ca²⁺, 10–12 pmol mg⁻¹ h⁻¹, accounted for the increased Ca²⁺ content after hatching. When comparing the measured and estimated ratios of efflux and influx, active transport was suggested to be involved in the uptake of Ca²⁺. Chloride cells, which may be responsible for the active uptake of Ca²⁺, started to differentiate in the skin of embryos 48h after fertilization, and the density of chloride cells increased following the development. A possibility of active transport for Ca²⁺ in early developmental stages of tilapia is suggested.     

Effects of sulfide on K+ flux pathways in red blood cells of crucian carp and rainbow trout

The effect of sulfide on K⁺ influx pathways was measured in red blood cells (RBCs) of sulfide-sensitive rainbow trout (Oncorhynchus mykiss) and sulfide-tolerant crucian carp (Carassius carassius). In trout RBCs, maximal inhibition of Na⁺, K⁺-ATPase was attained at 10 mol l^–1 sulfide and amounted to 32% without being influenced by pH between 6.7 and 8.3. Ouabain-resistant K⁺ influx in the absence and presence of sulfide was insignificant at pH values between 6.7 and 7.7. At higher pH values ouabain-resistant K⁺ influx increased, but was inhibited to about 15% by 30 mol l^–1 sulfide. In RBCs of crucian carp neither Na⁺, K⁺-ATPase nor ouabain-resistant K⁺ influx were affected by sulfide concentrations up to 850 mol l^–1. Differences in sulfide-sensitivity of K⁺ influx between both species can be based upon different properties of the membrane transporter themselves. The reduced Na⁺, K⁺-ATPase activity in trout RBCs may also result from a slightly reduced (by 9%) ATP level after sulfide exposure. In addition, intracellular sulfide concentrations were higher in trout RBCs as compared to crucian carp. In trout, intracellular sulfide concentrations reached extracellular levels within 5 min of incubation whereas sulfide concentrations in crucian carp RBCs remained about 2-fold lower than extracellular concentrations. Although the physiological basis of sulfide-insensitive K⁺ influx in crucian carp RBCs is currently unknown it may contribute to the extremely high sulfide-tolerance of this species.     

Intestinal HCO3− secretion in marine teleost fish: evidence for an apical rather than a basolateral Cl−/HCO3− exchanger

Intestinal fluid was collected from 11 marine teleost fish from the Baltic sea and the Pacific ocean. The anterior, mid and posterior segments of the intestine contained 33–110 mM of HCO₃ ^– equivalents (with exception of the Atlantic cod which contained only 5–15 mM). Considering literature values of transepithelial potentials and concentration gradients, these high levels of HCO₃ ^– equivalents are probably the result of active HCO₃ ^– transport. Possible HCO₃ ^– transport mechanisms were studied in the Pacific sanddab (Citharichthys sordidus) in vitro. Measurements of net secretion of HCO₃ ^– equivalents across the intestinal epithelium revealed mucosal DIDS sensitivity (10^–4 M) and Cl^–-dependence of the HCO₃ ^– equivalent net flux, but no serosal DIDS (10^–4 M) sensitivity. Net Na⁺ uptake was abolished in the absence of Cl^–, but some Cl^– uptake persisted in the absence of Na⁺, at a rate similar to that of net HCO₃ ^– secretion. Anterior, mid and posterior segments of the intestine performed similarly. These observations support the presence of an apical rather than a basolateral Cl^–/HCO₃ ^– exchanger and thus contrast the currently accepted model for intestinal HCO₃ ^– secretion. This apical Cl^–/HCO₃ ^– exchanger alone, however, is not sufficient for maintaining the observed HCO₃ ^– equivalents gradient in vivo. We suggest a coupling of cytosolic carbonic anhydrase, a basolateral proton pump and the apical Cl^–/HCO₃ ^– exchanger to explain the intestinal HCO₃ ^– transport.     

Acute Toxicity of Synthetic Pyrethroid Cypermethrin on the Common Carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) Embryos and Larvae

In this study, the toxic effects on the embryos and larvae of the common carp were used as a model to investigate the synthetic pyrethroid pesticide, cypermethrin, which contaminates aquatic ecosystems. Data obtained from the cypermethrin acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The control and eight test experiments were repeated five times. The number of dead embryos significantly increased in response to cypermethrin concentrations 0.0001, 0.001, 0.01, 0.1, 1, 2, 4 and 8 μg l⁻¹ (p<0.05 for each case). The 48 h LC₅₀ value (with 95% confidence limits) of cypermethrin for common carp embryos was estimated at 0.909 (0.256–5.074) μg l⁻¹. Dose–response decreases in hatching success were recorded as 87.4, 85.0, 80.2, 71.4, 56.3, 48.6, 38.8 and 23.5%, respectively. The lowest concentration of cypermethrin (0.0001 μg l⁻¹) produced a significant increase in the number of dead larvae compared to the control group (p<0.05). The number of dead larvae significantly increased with increasing cypermethrin concentrations exposed for 1–96 h (p<0.05). The highest concentration (8 μg l⁻¹) showed the highest larvae mortality. The 96 h LC₅₀ value (with 95% confidence limits) of cypermethrin for common carp larvae was estimated at 0.809 (0.530–1.308) μg l⁻¹. The results of the study suggest that low levels of cypermethrin in the aquatic environment may have a significant effect on the reproduction and development of carp.     

Characterization of transport Na+-ATPases in gills of freshwater tilapia

Branchial plasma membranes from the freshwater cichlid teleostOreochromis mossambicus (tilapia) contain two Na⁺-dependent ATPases: Na⁺/K⁺ ATPase, and an amiloride-sensitive ATPase which is postulated to operate as a Na⁺/H⁺ (–NH₄ ⁺) ATPase. It is suggested that both enzyme activities are located in the basolateral membrane system of the chloride cells. K⁺ has opposing effects on the two enzymes: it stimulates Na⁺/K⁺ ATPase and inhibits Na⁺/H⁺ (–NH₄ ⁺) ATPase activity. Na⁺/H⁺ ATPase appears more sensitive to NH₄ ⁺ at low concentrations than Na⁺/K⁺ ATPase and the stimulatory effect by NH₄ ⁺ ions on the first enzyme could be important in facilitating NH₄ ⁺ excretion by tilapia gills under physiological conditions.In vitro maximum stimulation by NH₄ ⁺ is similar for the two enzymes (200%). In contrast to Na⁺/K⁺ ATPase, Na⁺/H⁺ ATPase activity is inhibited by supra-physiological (>20 mM) concentrations of NH₄ ⁺.     

Osmoregulation in the mudskipper,Boleophthalmus boddaerti II. transepithelial potential and hormonal control

Boleophthalmus boddaerti submerged in 10%, 50% and 80% seawater (sw) for 7 days, had whole body transepithelial potentials (TEP) of 3.3, 18.3 and 22.9 mV, respectively. Hypophysectomy significantly decreased the TEP ofB. boddaerti and reversed the polarity of the TEP of the fish exposed to 10% sw.Hypophysectomy also significantly decreased the branchial Na⁺-K⁺ activated adenosine triphosphatase (Na⁺,K⁺-ATPase) activity but increased the activity of branchial HCO₃ ^–-Cl^– stimulated adenosine triphosphatase (HCO₃ ^–,Cl^–-ATPase) inB. boddaerti exposed to 10% sw. However, survival in 10% sw was not significantly impaired by hypophysectomy and no significant change in plasma osmolality and plasma Na⁺ and Cl^– concentrations was observed.Various doses of ovine-prolactin or salmon-prolactin were unable to restore the TEP of hypophysectomizedB. boddaerti in 10% sw to that of the sham-operated fish. However, cortisol increased TEP to a positive value in hypophysectomizedB. boddaerti, though it was still lower than the sham-operated control. Cortisol treatment also affected the plasma osmolality, plasma Na⁺ and Cl^– contents and branchial Na⁺,K⁺-ATPase and HCO₃ ^–,Cl^–-ATPase activities. Overall, the hormonal control of osmoregulation inB. boddaerti appeared to differ from that of other teleosts.     

Mechanisms of action of pituitary adenylate cyclase-activating polypeptide (PACAP) on growth hormone release from dispersed goldfish pituitary cells

The mechanisms of pituitary adenylate cyclase activating polypeptide (PACAP) action on goldfish growth hormone (GH) release were investigated by examining GH release responses from dispersed goldfish pituitary cells to a synthetic mammalian (m)PACAP₃₈ peptide. It was established that GH release stimulated by 2-h exposure to mPACAP₃₈ was concentration-dependent, attenuated by the PACAP receptor antagonist mPACAP_6–38, and subject to neuroendocrine modulation by somatostatin. Maximal mPACAP₃₈-stimulated GH release was not additive to the responses elicited by either the adenylate cyclase activator forskolin or the cyclic (c)AMP analog 8-bromo-cAMP. The GH responses to mPACAP₃₈, forskolin and 8-bromo-cAMP, either alone or in combination, were abolished by H89, a protein kinase A (PKA) inhibitor. SQ22536, an adenylate cyclase inhibitor, attenuated forskolin- and mPACAP₃₈-stimulated GH release. In contrast, mPACAP₃₈-stimulated GH release were additive to the responses to two protein kinase C (PKC) activators and unaffected by two PKC inhibitors. These results suggest that the stimulatory action of PACAP on GH secretion is mediated through a cAMP- / PKA-dependent mechanism, whereas the involvement of PKC appears unlikely. The ability of mPACAP₃₈ to further enhance maximal GnRH (PKC)-dependent GH release, but not dopamine D1 agonist (PKA)-dependent GH secretion, is consistent with this hypothesis. A possible involvement of Ca²⁺ in PACAP action is also suggested. Two inhibitors of voltage-sensitive Ca²⁺ channel reduced the GH responses to mPACAP₃₈ in static incubation; conversely, mPACAP₃₈ increased intracellular [Ca²⁺] in identified, single goldfish somatotropes.     

The effects of experimental anaemia on CO2 excretionin vitro in rainbow trout,Oncorhynchus mykiss

The effects of severe experimental anaemia on red blood cell HCO₃ ^– dehydrationin vitro were examined in rainbow trout,Oncorhynchus mykiss. After 5 days of anaemia (haematocrit=4.9±1.1%) induced by intraperitoneal injection of phenylhydrazine hydrochloride, fish displayed elevated arterial CO₂ tensions (anaemic PaCO₂=3.19±0.42 torrvs. control PaCO₂=1.35±0.17 torr) and a significant acidosis (anaemic pHa=7.73±0.04vs. control pHa=7.99±0.04). However, after 15–20 days of anaemia (hct=6.6±0.8%) induced by blood withdrawal, the arterial CO₂ tension was significantly lower than the control value, suggesting that physiological adjustments occurred within this time period to compensate for the lowered haematocrit. Compensation probably did not involve alterations in ventilation, which was unaffected by 5 days of anaemia (anaemic ;w=786±187 ml min^–1 kg^–1 vs. control ;w=945±175 min^–1 kg^–1), based on indirect Fick principle measurements.Potential adaptations to longer term anaemia at the level of the red blood cells were investigated using a radioisotopic HCO₃ ^– dehydration assay. Owing to the difference in haematocrits, the HCO₃ ^– dehydration rate for blood from anaemic fish was significantly lower than that for control fish following equilibration at the same CO₂ tension. This difference was eliminated when HCO₃ ^– dehydration rates were measured on blood samples adjusted to the same haematocrit, a result which implies that the intrinsic rate of CO₂ excretion at the level of the red blood cell was not up-regulated during anaemia. The difference was also eliminated by equilibrating the blood samples with CO₂ tensions appropriate for the group from which the sample was obtained,i.e., PCO₂=1.4 torr for control samples and PCO₂=3.2 torr for anaemic samples; each at the appropriate haematocrit. It is concluded that the elevated PaCO₂ helps to reset CO₂ excretion to the control level, but that some additional physiological adjustment occurs to lower the PaCO₂ after 15–20 days of anaemia.     

Modulation of calcium uptake in cadmium-pretreated tilapia (Oreochromis mossambicus) larvae

Tilapia larvae were exposed to 0 (control), 50 (50-Cd) or 100 (100-Cd) μg l^-1 cadmium for 4 days and then transferred to cadmium-free fresh water for 3 days of detoxification. Total length and weight, calcium influx and total body calcium and cadmium content were examined at various times during detoxification. All the groups grew normally with regards to total length and body weight. Within the first 12h of detoxification the 50- and 100-Cd exposed groups released cadmium at the similar rate of about 24 ng mg^-1 h^-1 (or 140 ng larva^-1 h^-1). Later, however, this rate declined to only 4–16% of the initial level. Calcium influx in the control group showed a 10–26% increase during the detoxification period. Calcium influx in the 50-Cd group increased by about 280% and reached it peak at 12h. Calcium influx in the 100-Cd group increased by 440% and did not peak until 24h after transfer. After peaking, the influxes in both 50- and 100-Cd groups declined to the level of control at the end of the experiment. Calcium contents in 50- and 100-Cd groups increased more rapidly than that in control group within first 24h of the detoxification period. However the rate of increase in calcium content in three groups was the same after 24h. The changes in calcium influx appeared to be correlated with those in calcium content, and these suggested that tilapia larvae regulate the mechanism of calcium balance to compensate for the reduced calcium level in the body. This revised version was published online in July 2006 with corrections to the Cover Date.     

In vivo calcitropic effect of some vitamin D compounds in the marine Antarctic teleost,Pagothenia bernacchii

The Antarctic notothenioid, Pagothenia bernacchii, were found to have plasma total and free calcium levels, plasma inorganic phosphate and whole body calcium efflux rates which were similar to those seen in other teleosts. But total bone calcium was lower than reported for other teleosts. A single injection of vitamin D₃ (5 ng g^–1 fish) increased plasma total and plasma free calcium and these increases were associated with an increase in whole body calcium efflux and bone calcification. Conversely, the same treatment with 1,25-(OH)₂-D₃ reduced plasma free calcium. This seco-steroid also increased the specific activity of ⁴⁵Ca in bone at 40h post-injection but did not significantly effect total bone calcium, plasma total calcium or whole body calcium efflux. 25-OH-D₃ at the same dose had no effect on any of the parameters tested and none of the seco-steroids tested had any effect on plasma total inorganic phosphate. These data show that both D₃ and 1,25-(OH)₃-D₃ can have calcitropic effects in this marine teleost and that these two forms of vitamin D can exert different effects within the same species.     

Growth and physiological changes in scaled carp and blue tilapia under behavioral stress in mono- and polyculture rearing using a recirculated water system

Growth performance, carcass composition, liver and blood parameters ofscaled carp (C), Cyprinus carpio, and blue tilapia (T),Oreochromis aureus, reared for eight weeks in twomonoculture (100%C and 100%T) and two polyculture (60%C–40%T and40%C-60%T) conditions were investigated. In polyculture 40%C–60%T bothspecies achieved the highest levels of specific growth rate and the lowestlevels of food conversion ratio and carcass lipid content. In addition, theyexhibited the highest values of plasma pO₂ and pH and the lowestvalues of plasma pCO₂, cholesterol and albumin, although thedifferences among treatments were not significant in the case of tilapia.Tilapia showed significantly lower plasma Cl^– levels than underthe other conditions. Carp in monoculture and tilapia in polyculture60%C–40%T had the lowest levels of specific growth rate and significantlyhigher levels of liver lipids and plasma triglycerides than in the other groups.In addition, carp in monoculture exhibited a significantly higher haematocritthan in polyculture. No significant variations among treatments were observedconcerning plasma cortisol, glucose, osmolality, Na⁺, K⁺,HCO₃ ^– andHCO₃ ^–/H₂CO₃ in either species.The combination of scaled carp and blue tilapia, in which blue tilapia were themain species, proved to be the best for both species. It was suggested thatgrowth and physiological changes under mono- and polyculture rearing, in anintensive system, seem to be as a result of a different state of stress relatedto fish behavior.     

Quantification of presumptive Na+/H+ antiporters of the erythrocytes of trout and eel

The presumptive Na⁺/H⁺ exchange sites of trout and eel erythrocytes were quantified using amiloride-displaceable 5-(N-methyl-N-[³H]isobutyl)-amiloride (³H-MIA) equilibrium binding to further evaluate the mechanisms of i) hypoxia-mediated modifications in the trout erythrocyte -adrenergic signal transduction system and ii) the marked differences in the catecholamine responsiveness of this system between the trout and eel. MIA was a more potent inhibitor of both trout apparent erythrocyte proton extrusion (IC₅₀ = 20.1 ± 1.1 mol l^–1, N = 6) activity (as evaluated by measuring plasma pH changes after addition of catecholamine in vitro) and specific ³H-MIA binding (IC₅₀ = 257 ± 8.2 nmol l^–1, N = 3) than amiloride, which possessed a proton extrusion IC₅₀ of 26.1 ± 1.6 mol l^–1 (N = 6) and a binding IC₅₀ of 891 ± 113 nmol l^–1 (N = 3). The specific Na⁺ channel blocker phenamil was without effect on adrenergic proton extrusion activity or specific ³H-MIA binding. Trout erythrocytes suspended in Na⁺-free saline and maintained under normoxic conditions possessed 37,675 ± 6,678 (N = 6) amiloride-displaceable ³H-MIA binding sites per cell (B_max, presumptive Na⁺/H⁺ antiporters) with an apparent dissociation constant (K_D) of 244 ± 29 nmol l^–1 (N = 6). Acute hypoxia (PO₂ = 1.2 kPa; 30 min) did not affect the K_D, yet resulted in a 65% increase in the number of presumptive Na⁺/H⁺ antiporters. Normoxic eel erythrocytes, similarly suspended in Na⁺-free saline, possessed only 17,133 ± 3,716 presumptive Na⁺/H⁺ antiporters (N = 6), 45% of that of trout erythrocytes, with a similar K_D (246 ± 41 nmol l^–1, N = 6). These findings suggest that inter- and intra-specific differences in the responsiveness of the teleost erythrocyte -adrenergic signal transduction system can be explained, in part, by differences in the numbers of Na⁺/H⁺ exchange sites.     

Effect of triiodothyronine on the growth and survival of larval striped bass (Morone saxatilis)

This study was carried out to test the effect of triiodothyronine (T₃) on the growth and survival of larval striped bass (Morone saxatilis). Growth and survival of striped bass held in 5 ppt seawater and treated with various doses of T₃ were measured beginning at 5 and 16 days after hatching. Body content of T₃ was measured by radioimmunoassay. T₃ dissolved in the 5 ppt seawater was taken up by larval striped bass in a dose-dependent manner, and affected the growth and survival of the fish. At 5 days after hatching, T₃ at 100 ng ml^–1 and 50 ng ml^–1 retarded the growth of larval striped bass and caused a lower survival rate than T₃ at 25 ng ml^–1 or the control treatment. At 16 days after hatching, T₃ at 100 ng ml^–1 retarded the growth of larval fish and caused a higher mortality. T₃ at 10 ng ml^–1 and 1 ng ml^–1 did not show any effect on either survival or growth. Body content of T₃ returns to control levels within days following end of treatment. The results indicate that exogenous T₃ can be detrimental to the growth and survival of larval striped bass.     

Changes in Na+,K+-ATPase activity and gill chloride cell morphology in the grouper Epinephelus coioides larvae and juveniles in response to salinity and temperature

The activity of the enzyme Na⁺,K⁺-ATPase and morphological changes of gill chloride cells in grouper, Epinephelus coioides larvae and juveniles were determined 6–48 h after abrupt transfer from ambient rearing conditions (30–32 ppt, 26.5–30 °C) to different salinity (8, 18, 32, 40 ppt) and temperature (25, 30 °C) combinations. Na⁺,K⁺-ATPase activity in day 20 larvae did not change at salinities 8–32 ppt. Activity decreased significantly (P <0.01) after exposure to 40 ppt at 25–30 °C, which was accompanied by an increase (P <0.05) in density and fractional area of chloride cells. Enzyme activity in 40 ppt did not reach a stable level and larvae failed to recover from an osmotic imbalance that produced a low survival at 25 °C and death of all larvae at 30 °C. Enzyme activity and chloride cell morphology in day 40 groupers did not change in 8–40 ppt at 25 °C and 8–32 ppt at 30 °C. A significant decrease and a subsequent increase in Na⁺,K⁺-ATPase activity in 40 ppt at 30 °C was associated with the increase in chloride cell density resulting in an increased fractional area but a decreased cell size. Enzyme activity and chloride cells of day 60 grouper were unaffected by abrupt transfer to test salinities and temperatures. These results demonstrate that grouper larvae and juveniles are efficient osmoregulators over a wide range of salinities. Salinity adaptation showed an ontogenetic shift as the larvae grew and reached the juvenile stage. This development of tolerance limits may reflect their response to actual conditions existing in the natural environment.     

Osmoregulation in the mudskipper,Boleophthalmus boddaerti I. Responses of branchial cation activated and anion stimulated adenosine triphosphatases to changes in salinity

The mudskipperB. boddaerti, was able to survive in waters of intermediate salinities (4–27). Fish submerged in dechlorinated tap water suffered 60% mortality by the fifth day while 60% of those in 100% sea-water (sw) died after the third day of exposure. After being submerged in 50% or 80% sw for 7 days, the plasma osmolality, plasma Na⁺ and Cl^– concentrations and the branchial Na⁺ and K⁺ activated adenosine triphosphatase (Na⁺,K⁺-ATPase) activity were significantly higher than those of fish submerged in 10% sw for the same period. However, the activities of the branchial HCO₃ ^– and Cl^– stimulated adenosine triphosphatase (HCO₃ ^–,Cl^–-ATPase) and carbonic anhydrase of the latter fish were significantly greater than those of the former. Such correlation suggests that Na⁺,K⁺-ATPase is important for hyperosmotic adaptation in this fish while HCO₃ ^–-Cl^–-ATPase and carbonic anhydrase may be involved in hypoosmotic survival.     

The effect of stocking density on growth,survival and agonistic behaviour of African catfish

A high rate of sibling cannibalism is one of the principal obstacles in the rearing of larvae and juveniles of the African catfish Clarias gariepinus. This paper examines the underlying behavioural components of agonistic behaviour under culture conditions. Ten day old catfish larvae were stocked at 30 larvae l^–1 in three tank designs with different surface areas and equal volumes. Stocking densities were 1.2, 0.6 and 0.3 fish cm^–2 bottom surface. Growth did not differ between treatments. The highest mortality over 30 days was recorded at the medium density (0.6 larvae l^–1). Fish stocked at this density showed the highest rate of aggression, while there was no difference in aggression between the highest and the lowest stocking densities. The results indicate that stocking density should be at least as high as 1.2 larvae cm^–2 bottom surface area to obtain high production at best survival rates.     

The conversion of plasma HCO 3 − to CO2 by rainbow trout red blood cells in vitro: adrenergic inhibition and the influence of oxygenation status

This study employed a recently developed radioisotopic assay (Wood and Perry 1991) to examine the inhibition, induced by catecholamines, of the conversion of plasma HCO ₃ ^– to CO₂ in acidotic trout blood, and the influence of oxygenation status on the response. Blood was incubated in vitro at P_{CO 2}= 2 torr, and 10^–6 M noradrenaline was employed as the adrenergic stimulus. In particular we investigated whether the inhibition of plasma HCO ₃ ^– conversion could be explained by a limited supply of H⁺s for the intracellular HCO ₃ ^– dehydration reaction because of competition by the adrenergically activated Na /H⁺ exchanger. Hypoxia (P_{O 2}= 15 torr) was employed as a tool to intensify this competition. Hypoxia raised RBC pHi, pHe, and plasma total CO₂ concentration (C_{CO 2}) by the Haldane effect, and increased the magnitude of Na⁺/H⁺ activation, expressed as the change in the transmembrane pH gradient (pHe-pHi). However hypoxia did not alter the inhibition of the conversion of plasma HCO ₃ ^– to CO₂ caused by noradrenaline. Hypoxia itself stimulated the RBC-mediated conversion of plasma HCO ₃ ^– to CO₂ by about 20% in the presence or absence of noradrenaline. The conversion rate was strongly correlated with pHe, pHe-pHi, and plasma C_{CO 2} in these experiments, but not with pHi. We conclude that adrenergically mediated inhibition in the conversion of plasma HCO ₃ ^– to CO₂ by trout RBCs is not due to competitive limitation on intracellular H⁺s, but rather to changes in the electrochemical gradient for HCO ₃ ^– entry and/or to CO₂ recycling from plasma to RBC. The deoxygenated condition helps to promote CO₂ excretion at the level of the RBC.     

Feeding early larval stages of fire shrimp Lysmata debelius (Caridea, Hippolytidae)

An important constraint to the commercial rearing of the marine ornamental shrimp Lysmata debelius is high larval mortality during early stages due to inappropriate procedures of larval collection and not feeding a live prey before one day elapsed after hatching. This incorrect feeding practice is commonly adopted in larval rearing of L. debelius and other ornamental marine shrimps because it is wrongly assumed that reserves of the newly hatched are enough for the first 24 h of life. Present work demonstrates that captive newly hatched L. debelius larvae ingest microalgae within minutes after hatching. When fed solely with Artemia nauplii, they have acceptable survival rates with stocking densities at or below 50 larval L^–1; but when nauplii are combined with microalgae, survival is further improved to zoea 2 as initial mortality is reduced, and higher stocking densities are supported (up to 75 larvae L^–1). The microalgae used were Rhinomonas reticulata, Skeletonema costata and Tetraselmis chuii. Higher survival through metamorphosis to zoea 2 was always observed for groups fed combinations of microalgae including Tetraselmis chuii. It is recommended that, larval collection methods ensure that larvae are fed microalgae within 2–3 h of release.     

Replacement of Artemia with Moina micrura in the rearing of freshwater shrimp larvae

The effect of an abrupt change in the live diet of shrimp larvae was investigated by replacing Artemia with Moina micrura. The control treatment consisted of feeding Artemia throughout the rearing period (regime A), while in the other treatments the onset of Moina feeding was arbitrarily chosen at larval stages iv (A3M), vi (A5M), viii (A7M) and x (A9M). No significant differences ( = 0.05) were observed among the treatments during larval production, mean stage development (MSD) and growth of postlarvae. The mean (SD) yields of postlarvae (PL) were 11.97 (1.98), 15.10 (2.92), 14.72(1.56), 13.51 (1.74) and 12.70 (1.40) PL l^–1 respectively for the feeding regimes A3M, A5M, A7M, A9M and A. Up to stage v, the ingestion rate in the Moina treatment was as low as 0.01–0.47 larva^–1 h^–1 compared with that in the Artemia treatment (0.29–1.77 larva^–1 h^–1). However, the ingestion of Moina increased from stage vi–vii onwards.