Effects of environmental salinity and temperature on osmoregulatory ability, organic osmolytes, and plasma hormone profiles in the Mozambique tilapia (Oreochromis mossambicus)

The Mozambique tilapia, Oreochromis mossambicus, is capable of surviving a wide range of salinities and temperatures. The present study was undertaken to investigate the influence of environmental salinity and temperature on osmoregulatory ability, organic osmolytes and plasma hormone profiles in the tilapia. Fish were acclimated to fresh water (FW), seawater (SW) or double-strength seawater (200% SW) at 20, 28 or 35 degrees C for 7 days. Plasma osmolality increased significantly as environmental salinity and temperature increased. Marked increases in gill Na(+), K(+)-ATPase activity were observed at all temperatures in the fish acclimated to 200% SW. By contrast, Na(+), K(+)-ATPase activity was not affected by temperature at any salinity. Plasma glucose levels increased significantly with the increase in salinity and temperature. Significant correlations were observed between plasma glucose and osmolality. In brain and kidney, content of myo-inositol increased in parallel with plasma osmolality. In muscle and liver, there were similar increases in glycine and taurine, respectively. Glucose content in liver decreased significantly in the fish in 200% SW. Plasma prolactin levels decreased significantly after acclimation to SW or 200% SW. Plasma levels of cortisol and growth hormone were highly variable, and no consistent effect of salinity or temperature was observed. Although there was no significant difference among fish acclimated to different salinity at 20 degrees C, plasma IGF-I levels at 28 degrees C increased significantly with the increase in salinity. Highest levels of IGF-I were observed in SW fish at 35 degrees C. These results indicate that alterations in gill Na(+), K(+)-ATPase activity and glucose metabolism, the accumulation of organic osmolytes in some organs as well as plasma profiles of osmoregulatory hormones are sensitive to salinity and temperature acclimation in tilapia.     

Heat shock protein (Hsp70) induced by a mild heat shock slightly moderates plasma osmolarity increases upon salinity transfer in rainbow trout (Oncorhynchus mykiss)

We have investigated whether mild heat shock, and resulting Hsp70 expression, can confer cross-protection against the stress associated with transfer from freshwater (FW) to seawater (SW) in juvenile rainbow trout (Oncorhynchus mykiss). In experimental Series I, juvenile trout reared in FW were transferred from 13.5 degrees C to 25.5 degrees C in FW, held for 2 h, returned to 13.5 degrees C for 12 h, and then transferred to 32 ppt SW at 13.5 degrees C. Branchial Hsp70 increased approximately 10-fold in the heat-shocked fish relative to the control by the end of recovery and remained high 2, 8, and 24 h post-salinity transfer. However, no clear differences could be detected in blood parameters (blood hemoglobin, hematocrit, MCHC, plasma Na(+) and plasma osmolarity) or muscle water content between heat-shocked and sham-shocked fish in SW at any sampling interval (0, 2, 8, 24, 48, 120, 240 and 360 h post-SW transfer). In experimental Series II, trout acclimated to 8 degrees C were heat-shocked at 22 degrees C for 2 h, allowed to recover 18 h, and exposed to a more severe salinity transfer (either 36 or 45 ppt) than in Series I. Branchial Hsp70 levels increased approximately 6-fold in heat-shocked fish, but had declined to baseline after 120 h in SW. Plasma osmolarity and chloride increased in both groups upon transfer to 36 ppt; however, the increase was significantly less in heat-shocked fish when compared to the increase observed in sham-shocked fish at 24 h. No significant differences could be detected in branchial Na(+)/K(+)-ATPase activity or Na(+)/K(+)-ATPase alpha1a and alpha1b mRNA expression between the two groups. Our data indicate that a mild temperature shock has only modest effects on the ability of rainbow trout to resist osmotic stress during FW to SW transfer.     

Stimulation of non-specific immune functions in seawater-acclimated rainbow trout, Oncorhynchus mykiss, with reference to the role of growth hormone

The influence of acclimation to seawater (SW) and growth hormone (GH) administration on immune functions was examined in the rainbow trout (Oncorhynchus mykiss). After 3 days acclimation to dilute SW (12 parts per thousand, ppt), an increase in plasma lysozyme activity was observed compared to the fish kept in fresh water (FW). No change was seen in plasma immunoglobulin M (IgM) levels. When they were transferred from dilute SW to full-strength SW (29 ppt) after a single intra-peritoneal injection of ovine or salmon GH, plasma sodium levels of GH-treated fish were significantly lower than those of the control fish injected with Ringer's solution 24 h after the transfer. The plasma level of IgM was not influenced by GH injection in the fish kept in FW nor in those transferred to SW. The administration of GH increased plasma lysozyme activity in the fish in FW, but no further increase was seen after SW transfer. The production of superoxide anions in peripheral blood leucocytes was stimulated by GH in both FW and SW. These results suggest that GH is involved in the stimulation of the non-specific immune functions in SW-acclimated salmonids.     

In vivo evidence for catecholaminergic inhibition of prolactin secretion in the teleostPoecilia latipinna

Summary Injections of L-dopa in freshwater (FW) fish reduced the size of the prolactin (PRL) cell nuclei, suggesting inhibition of PRL synthesis. A single injection of 6-hydroxydopamine (6-OHDA) in one-third seawater (1/3SW) fish reduced pituitary PRL content and increased PRL cell nuclear size at 6 h and 12 h, indicating stimulation of both synthesis and release of PRL. Two daily injections of 6-OHDA in 1/3SW fish led to PRL cell nuclear enlargement and elevated pituitary PRL content at 48 h after the second injection, indicating strong stimulation of PRL synthesis. Consideration of other parameters (plasma and body sodium levels, plasma osmotic pressure) suggests that the PRL cell responses to 6-OHDA were not mediated by internal osmotic changes. Pretreatment with 6-OHDA also appeared to accelerate the PRL cell activation induced by the transfer of fish from 1/3SW to FW.L-dopa opposed the enhancement of PRL release induced by a single injection of 6-OHDA. Injections of 3,4-dimethoxyphenylethylamine (DMPEA), a specific dopamine antagonist, caused short-term depletion of pituitary PRL, indicating enhanced PRL release.These results suggest that PRL secretion is subject to catecholaminergic inhibition, probably by dopamine. Considering these findings together with previous in vitro results (Wigham et al., 1975), it appears that the PRL cells are innervated by inhibitory catecholaminergic nerve fibres.Abbreviations DMPEA 3,4-dimethoxyphenylethylamine - L-dopa L-dihydroxyphenylalanine - FW freshwater - 6-OHDA 6-hydroxydopamine - PRL prolactin - 1/3SW one-third seawater     

Effect of pimozide on the cytology of the eel pituitary

Pimozide, a specific blocker of dopaminergic receptors, was injected for 4 to 9 days in freshwater (FW) eels or eels acclimated to sea water (SW), for 10 to 30 days. The daily dose was 100 or 200 microgram/100 g. In FW, pimozide induces a nuclear hypertrophy in the prolactin (PRL) cells of eels; these elongated cells increase in height. The amount of erythrosinophilic granules in the cytoplasm, initially reduced, increases. Plasma electrolyte values are not modified: only the plasma sodium level slightly rises with the higher dose. In SW, PRL cells appear less active. After 10 days, this hypoactivity is not yet fully evident; pimozide stimulates PRL cells without affecting electrolyte values. After 1 month in SW, PRL cells are stimulated with pimozide and a slight regranulation may occasionally occur. The response in SW is never as marked as it is in FW; a high dose is not more effective than a low one. The higher dose significantly raises Na+, Ca2+ and Cl- plasma levels. These data suggest that prolactin synthesis and release increase with pimozide. They corroborate the hypothesis of a hypothalamic inhibitory control on PRL secretion mediated through dopaminergic fibers in the eel, but other factors may also be involved in this regulation in addition to the effect of salinity.     

Plasma and erythrocyte solute properties of juvenile bull sharks, Carcharhinus leucas, acutely exposed to increasing environmental salinity

Plasma and erythrocyte solute properties were examined in freshwater (FW) acclimated juvenile Carcharhinus leucas following acute transfer to 75% seawater (SW), and 100% SW. Blood samples were taken at 0, 12 and 96 h following transfer to 75% SW and 24 and 72 h after transfer to 100% SW. A control group in FW was subjected to the same sampling regime. Upon transfer of C. leucas to 75% and 100% SW, plasma Na⁺, Cl⁻, K⁺, Mg²⁺, Ca²⁺, urea and TMAO concentrations all increased significantly but disproportionately. Plasma Na⁺ and Cl⁻ increased immediately, followed by an increase in plasma urea. Erythrocyte urea and TMAO concentrations increased significantly following transfer to 75% and 100% SW; however, changes in erythrocyte inorganic ion concentrations were insignificant. Haematocrit, haemoglobin and mean cell haematocrit did not differ significantly after transfer to seawater; however, plasma water was slightly reduced after 24 and 72 h in 100% SW. Red blood cell (RBC) water content was elevated 24 h after transfer to 100% SW but returned to FW levels after 72 h. These results demonstrate that the transfer from 75% to 100% SW presents C. leucas with a greater osmoregulatory challenge than transfer from FW to 75% SW, despite the larger concentration gradient in the latter. In summary, C. leucas tolerate rapid and significant increases in salinity by rapidly increasing plasma osmolality to be hyperosmotic to the environment whilst maintaining a tight regulation of their intracellular fluid environment.     

Osmoregulation and expression of ion transport proteins and putative claudins in the gill of southern flounder (Paralichthys lethostigma)

The southern flounder is a euryhaline teleost that inhabits ocean, estuarine, and riverine environments. We investigated the osmoregulatory strategy of juvenile flounder by examining the time-course of homeostatic responses, hormone levels, and gill Na(+),K(+)-ATPase and Na(+),K(+),2Cl(-) cotransporter protein expression after salinity challenge. Transfer of freshwater (FW)-acclimated flounder to sea water (SW) induced an increase in plasma osmolality and cortisol and a decrease in muscle water content, plasma insulin-like growth factor I (IGF-I) and hepatic IGF-I mRNA, all returning to control levels after 4 days. Gill Na(+),K(+)-ATPase and Na(+),K(+),2Cl(-) cotransporter protein levels were elevated in response to SW after 4 days. Transfer of SW-acclimated flounder to FW reduced gill Na(+),K(+)-ATPase and Na(+),K(+),2Cl(-) cotransporter protein, increased plasma IGF-I, but did not alter hepatic IGF-I mRNA or plasma cortisol levels. Gill claudin-3 and claudin-4 immunoreactive proteins were elevated in FW versus SW acclimated flounder. The study demonstrates that successful acclimation of southern flounder to SW or FW occurs after an initial crisis period and that the salinity adaptation process is associated with changes in branchial expression of ion transport and putative tight junction claudin proteins known to regulate epithelial permeability in mammalian vertebrates.     

Handling of ferric iron by branchial and intestinal epithelia of climbing perch (Anabas testudineus Bloch)

With a view to test how the branchial and intestinal tissues of fish, the two sites of metal acquisition, utilize the water-borne ferric [Fe(III)] iron and whether the accumulation of this form of iron influences cellular Na/K gradient in these tissues, the gills and intestines of climbing perch adapted to freshwater (FW) and acclimated to dilute seawater (20 ppt; SW) were analyzed for ouabain-sensitive Na+, K+-ATPase activity, Fe and electrolyte contents after loading a low (8.95 microM) or high dose (89.5 microM) of Fe(III) iron in the water. The SW gills showed higher levels of total Fe after treating with 8.95 microM of Fe(III) iron which was not seen in the FW gills. Na+, K+-ATPase activity, reflecting Na/K pump activity, showed an increase in the FW gills and not in the SW gills. Substantial increase in the branchial Na and K content was observed in the SW gills, but the FW gills failed to show such effects after Fe(III) loading. The total Fe content was declined in the FW intestine but not in the SW intestine. Water-borne Fe(III) iron decreased the activity of Na+, K+-ATPase in the SW intestine while not changing its activity in the FW intestine. The Na and K content in the FW intestine did not respond to Fe(III) iron exposure but showed a reduction in its Na levels in the SW intestine. The moisture content in the gills and intestines of both the FW and SW perch remained unaffected after Fe(III) loading. In FW fish, the plasma Na levels were decreased by a low dose of Fe(III) iron, though a high dose of Fe(III) iron was required in the SW fish for such an effect. Overall, the results for the first time provide evidence that gills act as a major site for Fe(III) iron absorption and accumulation during salinity acclimation which depends on a high cellular Na/K gradient.     

Osmoregulatory effects of hypophysectomy and homologous prolactin replacement in hybrid striped bass

The effects of ovine prolactin (oPRL) and striped bass prolactin (sbPRL; Morone saxatilis) on plasma osmolality, electrolyte balance, and gill Na(+),K(+)-ATPase activity were investigated in hypophysectomized (Hx), freshwater (FW)-acclimated, hybrid striped bass (M. saxatilisxMorone chrysops). They were kept in dilute (isoosmotic) seawater for about 10 days after surgery. Seven days after transfer to FW, Hx fish had lower plasma osmolality and lower levels of Na(+), Cl(-), and Ca(2+) than sham-operated and intact fish. Fish were injected four times with oPRL (1, 5, or 20 microg/g body mass), sbPRL (10 or 100 ng/g), or hormone vehicle (0.9% NaCl) at 48-h intervals (days 0, 2, 4, and 6) in FW and then sampled for blood plasma 24 h after the fourth injection (day 7). In Hx fish, oPRL (5 and 20 microg/g) and sbPRL (10 and 100 ng/g) were effective in maintaining plasma osmolality and levels of Na(+), Cl(-), and Ca(2+) above values seen in saline-injected controls. Hypophysectomy did not affect branchial Na(+),K(+)-ATPase activity, but enzyme activity was significantly reduced in Hx fish receiving oPRL (20 mug/g) or sbPRL (10 or 100 ng/g). These results indicate that PRL acts to maintain plasma osmotic and ionic balance in FW-adapted hybrid striped bass, and that this may involve downregulation of branchial Na(+),K(+)-ATPase activity.     

Prolactin regulation in the coho salmon,Oncorhynchus kisutch

Summary Parr and smolt sea water acclimated coho salmon,Oncorhynchus kisutch were subjected to gradual and direct transfers to fresh water. Plasma osmotic pressure, Na⁺, K⁺, Ca⁺⁺ and Mg⁺⁺ were similar in freshwater (FW) fish and seawater (SW) transferred controls for the 24 h following transfer. In spite of the similarity in osmotic pressure and ion levels, plasma cortisol concentrations were significantly increased immediately following salinity change while both pituitary and plasma prolactin decreased indicating enhanced secretion by the pituitary and clearance from the blood. In vitro experiments showed greater incorporation of tritiated leucine into prolactin (PRL) cells immediately after transfer to FW while prolactin injections into intact fish lowered activity in rostral pars distalis (RPD) cells as measured by the same technique, providing evidence of hormonal feedback. These experiments show that the increased synthesis and release of PRL that occurs in coho following movement into FW is not obviously correlated with plasma osmotic pressure, Na⁺ or Ca⁺⁺ concentrations as has been observed in other species of teleosts.Abbreviations FW freshwater - SW seawater - PRL prolactin - RPD rostral pars distalis     

Expression and distribution of Na, K-ATPase in gill and kidney of the spotted green pufferfish, Tetraodon nigroviridis, in response to salinity challenge

Freshwater (FW) spotted green pufferfish (Tetraodon nigroviridis) were transferred directly from a local aquarium to fresh water (FW; 0 per thousand ), brackish water (BW; 15 per thousand ), and seawater (SW; 35 per thousand ) conditions in the laboratory and reared for at least two weeks. No mortality was found. To investigate the efficient mechanisms of osmoregulation in the euryhaline teleost, distribution and expression of Na,K-ATPase (NKA) in gill and kidney of the pufferfish were examined and the osmolality, [Na+] and [Cl-] of the blood were assayed. The lowest levels of both relative protein abundance and activity were found to be exhibited in the BW group, and higher levels in the SW group than FW group. In all salinities, branchial NKA immunoreactivity was found in epithelial cells of the interlamellar region of the filament and not on the lamellae. Relative abundance of kidney NKA alpha-subunit, as well as the NKA activity, was found to be higher in the FW pufferfish than fish in BW or SW. Renal NKA appeared in the epithelial cells of distal tubules, proximal tubules, and collecting tubules, but not in glomeruli, in fish groups of various salinities. Plasma osmolality and chloride levels were significantly lower in FW pufferfish than those in BW and SW, whereas plasma sodium did not differ among the groups. Although identical distributions of NKA were found in either gill or kidney of FW-, BW- or SW-acclimated spotted green pufferfish, differential NKA expression in fish of various salinity groups was associated with physiological homeostasis (stable blood osmolality), and illustrated the impressive osmoregulatory ability of this freshwater and estuarine species in response to salinity challenge.     

Ionoregulatory and endocrine responses to disturbed salt and water balance in Mozambique tilapia exposed to confinement and handling stress

This study assessed the endocrine and ionoregulatory responses by tilapia (Oreochromis mossambicus) to disturbances of hydromineral balance during confinement and handling. In fresh water (FW), confinement and handling for 0.5, 1, 2 and 6 h produced elevations in plasma cortisol and glucose; a reduction in plasma osmolality was observed at 6 h. Elevations in plasma prolactins (PRL₁₇₇ and PRL₁₈₈) accompanied this fall in osmolality while no effect upon growth hormone (GH) was evident; an increase in insulin-like growth-factor I (IGF-I) occurred at 0.5 h. In seawater (SW), confinement and handling increased plasma osmolality and glucose between 0.5 and 6 h; no effect on plasma cortisol was seen due to variable control levels. Concurrently, both PRLs were reduced in stressed fish with only transient changes in the GH/IGF-I axis. Next, the branchial expression of Na⁺/K⁺/2Cl^? cotransporter (NKCC) and Na⁺/Cl^? cotransporter (NCC) was characterized following confinement and handling for 6 h. In SW, NKCC mRNA levels increased in stressed fish concurrently with elevated plasma osmolality and diminished gill Na⁺, K⁺-ATPase activity; NCC was unchanged in stressed fish irrespective of salinity. Taken together, PRL and NKCC participate in restoring osmotic balance during acute stress while the GH/IGF-I axis displays only modest responses.     

Extracellular ionic and acid-base adjustments of Atlantic salmon presmolts and smolts in fresh water and after transfer to sea water: the effects of ovine growth hormone on the acquisition of euryhalinity

In order to better understand the basis for the acquisition of euryhalinity by juvenile salmon and the role of endogeneous stimuli, experiments have been carried out to examine the dynamics of ionic and acid-base adjustments in fresh water (FW) and after direct transfer to full salinity (32 g l⁻¹) sea water (SW) (1) on Atlantic salmon smolt during the natural period of smoltification in spring, (2) on presmolt salmon in autumn, after intraperitoneal implantation of pellets containing ovine growth hormone (oGH). During parr-smolt transformation in FW, gill Na⁺/K⁺ ATPase activity gradually rises, the plasma osmolality (Posm) is unaffected and the total CO₂ of the plasma decreases significantly while whole blood pH fluctuates slightly. Direct transfer of smolt from FW to SW provokes only a slight increase in Posm and emphasizes the acid-base balance disruptions shown in FW. An oGHimplant in a presmolt stimulates gill Na⁺/K⁺ ATPase activity in FW, and affects the acid-base balance. After SW transfer (12 days after implantation), oGH treatment prevents the increase of osmotic pressure and the restoration of the acid-base, ionic equilibrium was faster for oGH-implanted fish than for sham-operated fish. These observations show that in FW smelting salmon develop most of the systems they need for migration and growth in SW and that oGH implants induce the development of physiological characteristics of smolts in a non-natural period of smolting.     

Stanniectomy attenuates the renin–angiotensin response to hypovolemic hypotension in freshwater eels (<Emphasis Type="Italic">Anguilla rostrata</Emphasis>) but not blood pressure recovery

Plasma Angiotensin II (ANG II) concentrations were measured in SHM (sham-operated 2 weeks) and CSX (corpuscles of Stannius removed 2 weeks) eels before and after the induction of hypovolemic hypotension (HH) by the rapid withdrawal of 8 ml kg bw⁻¹ of caudal venous blood. Baseline (before exsanguination) plasma ANG II concentrations were similar in SHM and CSX eels (81.3 ± 18.8 fmol ml⁻¹ cf. 106 ± 31.6 fmol ml⁻¹, respectively) but the elevation in plasma ANG II following HH (1,732 ± 82 fmol ml⁻¹) was attenuated by CSX (368 ± 127 fmol ml⁻¹) showing that the CS are linked to plasma ANG II concentrations. Plasma ANG II in both groups returned to baseline levels within 48 h. Dorsal aortic blood pressures (DABP) were measured in both experimental groups before, and during the 60 min after, blood withdrawal. A 44% decrease in mean DABP was observed in both SHM and CSX eels within 2 min and followed by similar rapid patterns of recovery of systolic, diastolic, and pulse pressures in both groups during the next 60 min showing that short-term recovery of DABP is not CS-dependent. Stanniectomy increased plasma Ca and K⁺ and decreased plasma Mg, Na⁺, Cl⁻ and osmolality which confirms some earlier observations in eels and other freshwater teleosts.     

Response of prolactin cells to environmental calcium in the eel (Anguilla anguilla L.)

Prolactin (PRL) cell activity was investigated in eels kept in fresh water (FW), deionized water (DW) supplemented or not with Ca (2 mM), in Ca-enriched FW (10 mM), in normal (Ca 3.4 mM) or Ca-free 1/3 sea water (SW), and in SW (Ca 10.2 mM) or Ca-free SW (Ca 0.15 mM). Light-microscopic studies, including measurement of the nuclear area and cell height, showed that PRL cell activity, reduced in DW, is not affected by Ca supplementation. Activity is reduced in Ca-enriched FW, in 1/3 SW and in SW, conditions inducing an increase in the plasma sodium level. The lack of calcium in saline environments partly suppresses the nuclear atrophy occurring in SW. There is no significant correlation between external or total plasma calcium concentration and PRL cell activity. In artificial Ca-free SW, eels show a rapid increase in plasma osmolarity and sodium levels; there is a significant negative correlation between these two plasma values and the nuclear area or cell height of PRL cells. As in some other teleosts, plasma osmolarity and plasma sodium seem to play a more important role than external or internal calcium in controlling PRL secretion. This correlation is not apparent in eels kept in SW, having unstimulated PRL cells but active calcium-sensitive (Ca-s) cells in the pars intermedia.     

IGF-I and branchial IGF receptor expression and localization during salinity acclimation in striped bass

The initial response of the IGF-I system and the expression and cellular localization of IGF type-I receptor (IGF-IR) were studied in the gill of a euryhaline teleost during salinity acclimation. Exposure of striped bass (Morone saxatilis) to hyperosmotic and hypoosmotic challenges induced small, transitory (<24 h) deflections in hydromineral balance. Transfer from freshwater (FW) to seawater (SW) induced an initial decrease in plasma IGF-I levels after 24 h in both fed and fasted fish. There was an overall decrease in liver IGF-I mRNA levels after SW transfer, suggesting that decreased plasma levels may be due to a decline in hepatic IGF-I synthesis. No changes were observed in gill IGF-I mRNA, but SW transfer induced an increase in gill IGF-IR mRNA after 24 h. Transfer from SW to FW induced an increase in plasma IGF-I levels in fasted fish. In fed fish, no significant changes were observed in either plasma IGF-I, liver, or gill IGF-I mRNA, or gill IGF-IR mRNA levels. In a separate experiment, FW-acclimated fish were injected with saline or IGF-I prior to a 24-h SW challenge. Rapid regain of osmotic balance following SW transfer was hindered by IGF-I. Immunohistochemistry revealed for the first time in teleosts that IGF-IR and Na(+)-K(+)-ATPase are localized in putative chloride cells at the base of the lamellae, identifying these cells in the gill as a target for IGF-I and IGF-II. Overall the data suggest a hyperosmoregulatory role of IGF-I in this species.     

Effects of dietary trimethylamine on free amino acid and nonprotein nitrogen levels in muscle of the guppy, Poecilia reticulata, in relation to seawater adaptation

Guppies Poecilia reticulata acclimated to 100% seawater (SW) had lower taurine and alanine levels in muscle than fish kept in freshwater (FW). The glycine level, in contrast, was higher in SW fish than in FW fish. Levels of other free amino acids (FAA) were comparatively low and little different between fish adapted in FW and in SW. In both FW and SW fish almost all of muscle FAA showed little difference in levels between fish kept on diets containing three different levels of trimethylamine (TMA) (0, 223, and 334 mumol TMA/kg dry weight of diet). Total FAA and nonprotein nitrogen levels in muscle were unaffected by the difference in either the diet species or the ambient salinities. Muscle trimethylamine oxide levels were higher in SW fish than in FW fish. In both salinities, muscle trimethylamine oxide levels in fish on the diets containing 223 and 334 mumol TMA/kg were slightly greater than the level in fish on the TMA-free diet.     

Changes to chloride and rodlet cells in gills, kidney and intestine of Dicentrarchus labrax (L.) exposed to reduced salinities

The effect of osmotic shock was investigated mainly in the chloride cells (CCs) and rodlet cells (RCs) of gills, and RCs of intestine and kidney of the European sea bass Dicentrarchus labrax obtained from a farm in the northern Adriatic Sea. During the experiment, fish were abruptly transferred from sea water to a salinity of 15 (15 SW) or to fresh water (FW). Numeric variation and ultrastructural changes of both cell types were evaluated at 24, 48 and 96 h after the transfer to lower salinity levels, using light and transmission electron microscopy (TEM). Exposure to FW produced a significant increase ( P < 0·05) in the number of branchial CCs and RCs within 96 and 24 h, respectively. Following osmotic challenge (either transfer to 15 SW or FW), kidney and intestine showed an evident increase in RC numbers. The cellular damage detected by TEM was the same for each sampling time (24, 48 and 96 h), but appeared more severe in fish exposed to FW (higher osmotic shock) than in those exposed to 15 SW. In RCs cytoplasmic vacuolizations, autophagosomes and autophagolysosomes with myelinoid bodies, dissolution and shrinkage of the typical inclusions were documented. Nevertheless, CCs showed vacuolization of endoplasmic reticulum and cytoplasmic dissolution and maintained the apical crypt typical of seawater acclimated fish. Renal tubular cells and intestinal epithelial cells showed similar changes to those reported for CCs and RCs.     

Time course of extracellular acid-base adjustments under hypo- or hyperosmotic conditions in the euryhaline fish Platichthys flesus

In order to better understand the basis for the euryhalinity of the flounder, Platichthys flesus , which tolerates large variations in water salinity, experiments have been designed to characterize the time course of extracellular ionic and acid-base adjustments under hypo- or hyperosmotic conditions. Abrupt transfer from sea water (SW) to fresh water (FW) provokes a transient decrease in the plasma osmolality (Posm) and a concomitant transient metabolic alkalosis (whole blood pH 7.78 in SW and 8.04 five days after FW transfer) associated with a marked, persistent hypercapnia. After 33 days in FW, Posm and whole blood pH are not significantly different from those in SW, but whole blood Pco₂ and plasma bicarbonate concentration are always higher than SW values. Opposite transitory fluctuations, i.e. a metabolic acidosis associated with a respiratory alkalosis, occur when flounder long-acclimated to FW are again exposed to SW. The mechanisms involved in these salinity-dependent acid-base disturbances are rather complex and remain to be elucidated. These observations attest to the importance of the extracellular acid-base changes that may be (i) linked to extracellular anisosmotic regulation and/or to cellular metabolic adjustments, and (ii) compensated partially by ventilatory adjustments.