Chemical composition of the body fluids andmuscle of the hagfish Myxine glutinosa and the rabbit-fish Chimaera monstrosa

Comprehensive analyses have been made of the plasma and parietal muscle of the marine cyclostome Myxine glutinosa and of the chimaeraoid Chimaera monstrosa , and the sum of some 17 constituents compared with the directly determined osmotic concentration of plasma. Both Myxine and Chimaera plasmas are isosmotic with sea water within 2%. Ions account for at least 96% of the total concentration of the plasma in Myxine and about 60% in Chimaera, but the blood of the latter contains 332 mM urea, compared with 3 mM in Myxine . In muscle a similar divergence exists, the urea concentrations per kilogram muscle water being practically the same as in the respective plasmas of the two species. The mean values in millimoles of the other nitrogenous constituents of muscle are for Myxine and Chimaera (figures for latter in brackets), trimethylamine oxide 87 (189), betaine 65 (38), creatine and creatinine 30 (37), amino acids 291 (43). Using osmotic coefficients the calculated osmotic concentration of muscle exceeds that of plasma in Myxine and possible reasons for this are discussed.
Estimates of inulin space in Myxine muscle have enabled intracellular concentrations of ions to be calculated. Analysis of Myxine bile shows it to have much higher concentrations of sodium, calcium and magnesium, and much lower chloride, than plasma. The resulting large anion deficit is presumably made up chiefly by the bile salt myxinol disulphate.     

Renal response of euryhaline toad (Bufo viridis) to acute immersion in tap water, NaCl, or urea solutions

Green toads (Bufo viridis) were acclimated to either tap water, 230 mOsmol NaCl kg-1 H2O (saline), 500 mOsmol NaCl kg-1 H2O (high saline), or 500 mmol L-1 urea. Renal functions for each acclimation group were studied on conscious animals that had one ureter chronically catheterized. Reciprocal immersion of tap-water- and saline-acclimated toads in the opposite solution did not stress the animals osmotically, and plasma osmolality increased or decreased by no more than 15%. However, urine osmolality and ionic composition changed immediately and profoundly on exposure to the other solution. Exposure of tap-water-acclimated toads to saline decreased urine flow by 30%, whereas the reciprocal immersion led to an increase of 30%. Immersion of tap-water-acclimated toads in high saline led to immediate cessation of urine flow, whereas immersion of 500 NaCl- or urea-acclimated toads in tap water led to a large increase in urine flow, with an overshoot that lasted 10 h (as a result of either salt or urea diuresis). Urine flow then stabilized at a level 5-6 times higher than the value attained at high-salt environment. On immersion of 500 urea-acclimated toads in 500 NaCl, urine flow doubled, accompanied by a change in ion composition, without change in the osmolality. In all experimental conditions, plasma potassium concentration was maintained within a narrow range. The results show that the toad's kidneys contributed efficiently both to osmo- and ionoregulation in a wide range of ambient solutions.     

THE EMERGENCE OF LARVAE FROM CYSTS OF THE BEET EELWORM, HETERODERA SCHACHTII SCHMIDT, IN AQUEOUS SOLUTIONS OF ORGANIC AND INORGANIC SUBSTANCES

The rate of larval emergence from cysts of the beet eelworm in a variety of aqueous solutions containing organic and inorganic substances is significantly higher than the emergence rate in water. It is suggested that differences between larval emergence rates in monoamino-monocarboxylic amino-acids may be related to the lipid solubility of these substances and their ability to penetrate the egg membranes. The larval emergence rate in fructose, glucose, sucrose and maltose was significantly higher than that in water, but in raffinose, arabinose and xylose the rate of emergence was no higher than in water. A high rate of larval emergence occurred in sodium chloride, potassium chloride and mercuric chloride, but not in magnesium chloride or calcium chloride. Experiments with several other organic solutions are described. There is an optimum concentration for larval emergence in beet diffusate. The osmotic pressure of the diffusate when maximum emergence occurred was 0·48 atm. Measurements of shrinkage of unhatched larvae in various concentrations of urea, sodium chloride and sucrose showed that decreasing rates of emergence at higher concentrations may be due to changes in the unhatched larvae brought about: by an osmotic effect. High concentrations of beet diffusate may have a similar effect.     

Exchanges of sodium and chloride at low salinities by Nereis diversicolor (Annelida, Polychaeta).

1. Experiments to compare the exchange (total influx) of sodium and chloride in the polychaete Nereis diversicolor in steady-state adaptation to very low salinities are reported. 2. The Na-uptake mechanism shows a high affinity for sodium, reaching half the maximal uptake rate at an external Na-concentration of 8-10 mM/liter (ca. 2% SW), and becomes "saturated" or reaches a plateau of uptake at concentrations of 40-50 mM/liter (ca. 10% SW) up to ca. 350 mM/liter (75% SW), above which Na-exchange is proportional to the external concentration. 3. The Cl-uptake curve differs from the Na-uptake curve in showing a relative depression at very low salinities before reaching "saturation" at Cl-concentrations of 50-60 mM/liter (ca. 10% SW). Cl-uptake becomes proportional to external concentration in salinities of 50% SW or greater, suggestive of passive diffusion in the ionic and osmotic conforming range. 4. It is shown that the permeability of the body wall, both to Na and to Cl, is reduced at very low salinities, thus destroying one of the assumptions upon which a previously-presented balance-sheet for chloride exchanges in N. diversicolor was based (Smith, 1970a). 5. Attempts to demonstrate an activation of the Na-uptake mechanism at very low salinities were inconclusive; reduction of body-wall permeability to sodium masks any possible activation. 6. It is suggested that the inside-negative body-wall potential is related to the depression of the Cl-uptake curve in salinities below 10% SW.     

Morphological responses of mitochondria-rich cells to hypersaline environment in the Australian mudskipper, Periophthalmus minutus

A population of the Australian mudskipper, Periophthalmus minutus, was found to inhabit mudflat that remained uncovered by tide for more than 20 days in some neap tides. During these prolonged emersion periods, P. minutus retreated into burrows containing little water, with a highest recorded salinity of 84 ± 7.4 psu (practical salinity unit). To explore the mechanical basis for this salinity tolerance in P. minutus, we determined the densities of mitochondria-rich cells (MRCs) in the inner and outer opercula and the pectoral fin skin, in comparison with P. novaeguineaensis, from an adjacent lower intertidal habitat, and studied morphological responses of MRCs to exposure to freshwater (FW), and 100% (34-35 psu) and 200% seawater (SW). Periophthalmus minutus showed a higher density of MRCs in the inner operculum (3365 ± 821 cells mm(-2)) than in the pectoral fin skin (1428 ± 161) or the outer operculum (1100 ± 986), all of which were higher than the MRC densities in p. novaeguineaensis. No mortality occurred in 100% or 200% SW, but half of the fish died within four days in FW. Neither 200% SW nor FW exposure affected MRC density. Transfer to 200% SW doubled MRC size after 9-14 days with no change in the proportion of MRCs with apical pits or plasma sodium concentration. In contrast, transfer to FW resulted in a rapid closing of pits and a significant reduction in plasma sodium concentration. These results suggest that P. minutus has evolved morphological and physiological mechanisms to withstand hypersaline conditions that they may encounter in their habitat.     

Plasma biochemistry changes in thoroughbred foals during the first 4 weeks of life.

Changes in plasma sodium, potassium, chloride, total carbon dioxide, urea, creatinine, glucose, total bilirubin, iron, total protein, albumin, alkaline phosphatase (AP), aspartate amino transferase (AST), calcium, inorganic phosphorus, cholesterol and triglycerides were studied in 45 Thoroughbred foals 15 min to 28 days after birth. The results were analysed in 3 groups; Group 1 (0--12 h), Group 2 (12--36 h), Group 3 (1--4 weeks). When Group 2 was compared to Group 1, there were significant reductions of sodium, creatinine, iron and calcium and elevations of total protein and bilirubin. When Group 3 was compared to Group 1 there were significant reductions of sodium, chloride, urea, creatinine, bilirubin, iron and AP. Significant elevations occurred in glucose, total protein, AST, inorganic phosphorus and triglycerides.     

Effects of dehydration on plasma osmolality, thirst-related behavior, and plasma and brain angiotensin concentrations in Couch's spadefoot toad, Scaphiopus couchii

Under dehydrating conditions, many terrestrial vertebrates species exhibit increases in plasma osmolality and their drinking behavior. Under some circumstances, this behavioral change is accompanied by changes in plasma and central angiotensin concentrations, and it has been proposed that these changes in angiotensin levels induce the thirst-related behaviors. In response to dehydration, the spadefoot toad, Scaphiopus couchii, exhibits thirst-related behavior in the form of cutaneous drinking. This behavior has been termed water absorption response (WR) behavior. Spadefoot toads live in harsh desert environments and are subject annually to dehydrating conditions that may induce thirst-related behavior. We tested the hypothesis that an increase in WR behavior is associated with both an increase in plasma osmolality and an increase in plasma and brain angiotensin concentrations. First, we determined the degree of dehydration that was necessary to initiate WR behavior. Animals dehydrated to 85% of their standard bladder-empty weight via deprivation of water exhibited WR behavior more frequently than control toads left in home containers with water available. Next, using the same dehydration methods, we determined the plasma osmolality and sodium concentrations of dehydrated toads. Toads dehydrated to 85% standard weight also had a significant increase in plasma osmolality, but exhibited no overall change in plasma sodium concentrations, indicating that while an overall increase in plasma osmolality appears to be associated with WR behavior in S. couchii, changes in sodium concentrations alone are not sufficient to induce the behavior. Finally, plasma and brain angiotensin concentrations were measured in control toads and toads dehydrated to 85% standard weight. Plasma and brain angiotensin concentrations did not increase in dehydrated toads, indicating that dehydration-induced WR behavior that is associated with changes in plasma osmolality may not be induced by changes in endogenous angiotensin concentrations in S. couchii.     

Plasma osmolyte concentrations and rectal gland mass of bull sharks Carcharhinus leucas, captured along a salinity gradient

Bull sharks (Carcharhinus leucas) were captured across a salinity gradient from freshwater (FW) to seawater (SW). Across all salinities, C. leucas were hyperosmotic to the environment. Plasma osmolarity in FW-captured animals (642 +/- 7 mosM) was significantly reduced compared to SW-captured animals (1067 +/- 21 mosM). In FW animals, sodium, chloride and urea were 208 +/- 3, 203 +/- 3 and 192 +/- 2 mmol l(-1), respectively. Plasma sodium, chloride and urea in SW-captured C. leucas were 289 +/- 3, 296 +/- 6 and 370 +/- 10 mmol l(-1), respectively. The increase in plasma osmolarity between FW and SW was not linear. Between FW (3 mosM) and 24 per thousand SW (676 mosM), plasma osmolarity increased by 22% or 0.92% per 1 per thousand rise in salinity. Between 24 per thousand and 33 per thousand, plasma osmolarity increased by 33% or 4.7% per 1 per thousand rise in salinity, largely due to a sharp increase in plasma urea between 28 per thousand and 33 per thousand. C. leucas moving between FW and SW appear to be faced with three major osmoregulatory challenges, these occur between 0-10 per thousand, 11-20 per thousand and 21-33 per thousand. A comparison between C. leucas captured in FW and estuarine environments (20-28 per thousand ) in the Brisbane River revealed no difference in the mass of rectal glands between these animals. However, a comparison of rectal gland mass between FW animals captured in the Brisbane River and Rio San Juan/Lake Nicaragua showed that animals in the latter system had a significantly smaller rectal gland mass at a given length than animals in the Brisbane River. The physiological challenges and mechanisms required for C. leucas moving between FW and SW, as well as the ecological implications of these data are discussed.     

Adaptation of trout to seawater. Effects on sodium plasma concentration, gill exchange and intestinal transport]

10. Trout Salmo irideus are adapted to sea water (S W) within four weeks by submitting them to a stepwise increase in external salinity (successively: 1/3 SW, 1/2 SW, 3/4 SW, full SW). 20. In completely adapted individuals mean plasma electrolyte concentrations vary only slightly (by a few %) from one medium to another. Thus, the trout may be regarded as a euryhaline, eventually homeosmotic species. 30. With increasing outside salinity there is a progressive diminution in the overall gill permeability to ions which is suggested by saturation curves obtained for sodium fluxes (maximum at about 500 muEq/h. 100 g at 15 degrees C, for unshocked fish). 40. Disturbance of the animals provokes a striking elevation and desequilibrium in these exchanges and this in turn induces an abnormal rise in plasma concentrations and a subsequent failure to adapt to hypertonic media. 50. In vitro absorption of water and sodium by intestinal everted-sacs increases only after transfer to full sea water. Mucosal entry of ions into intestinal epithelial cells measured by the technique of Schultz et al. (1967), is diminished in sea water-adapted animal (by 42% in the case of sodium). 60. These results demonstrate that Salmo irideus possesses efficient osmoregulatory mechanisms which operate with minimal energy expenditure in hypertonic media.     

Comparative study of selected blood biochemical components in milk or milk-replacer fed calves during the second week of life

The experiment was carried out on 13 male Polish Black and White dairy calves of 75% share of the Holstein-Friesian (HF) breed during the second week of life. The animals were divided into two groups. One group (n=7) was fed mother's milk and the second (n=6) milk replacer. The dynamics of changes in concentration of selected blood biochemical components connected with nitrogen metabolism (plasma total protein, albumin, urea, endogenous creatinine) and with mineral metabolism (sodium, chloride, calcium, magnesium, zinc, copper and plasma osmotic pressure) were analyzed in both groups. The results show that the type of ingested food influences the concentration of indicators reflecting nitrogen metabolism. Changes of these parameters in calves fed milk replacer are possibly connected with advantageous catabolic changes. Stable concentrations of main extracellular fluid electrolytes and blood plasma osmotic pressure were found in both groups of calves. Constant blood plasma calcium, magnesium, zinc and copper concentrations observed during this study might also indicate the relative maturity of mechanisms maintaining water and electrolyte balance. Nevertheless, it seemsjustifiable to monitor the copper concentrations in plasma of young calves.     

Plasma thyroxine and cortisol profiles and gill and kidney Na+/K+-ATPase and SDH activities during acclimation of the catfish Heteropneustes fossilis (bloch) to higher salinity, with special reference to the effects of exogenous cortisol on hypo-osmoregulatory ability of the catfish

When the stenohaline catfish Heteropneustes fossilis was transferred from fresh water (FW) to 30% seawater (SW), the Na(+)/K(+)-ATPase activity significantly increased in the kidney, while in gills it remained more or less constant. A reverse pattern was observed for succinic dehydrogenase (SDH) activity inasmuch as it significantly increased in gills and remained unchanged in the kidney. Plasma osmolality significantly increased within 3 days of transfer to 30% SW and remained significantly higher throughout the duration of experiment. These results suggest that catfish gills may not be able to reverse their function from salt uptake in FW to salt excretion at higher salinity, and that the elimination of monovalent as well as divalent ions is performed by the kidney but not the gills. The significant decline in plasma cortisol (F) levels following transfer to higher salinity may not be due to reduced production but rather to an enhanced utilization and clearance rate, a conclusion supported by the fact that exogenous administration of cortisol acetate (FA) resulted in significant increases in branchial and renal Na(+)/K(+)-ATPase in FW and 30% SW. FA also improved the plasma osmotic regulatory ability of the catfish, possibly due to a change in branchial function from salt-absorption to salt excretion, as was evident from a significant increase in branchial Na(+)/K(+)-ATPase activity in the fish in 30% SW pretreated with FA for 5 days. Consistently higher levels of plasma thyroxine (T4) following transfer to higher salinity suggest the involvement of this hormone at higher salinity.     

The anal portion as a salt-excreting organ in a seawater mosquito larva,A?des togoi Theobald

Summary Larvae of the marine mosquito,Aëdes togoi Theobald, tolerate environmental salinities ranging from fresh water to 300% sea water. When they were transferred from fresh water to sea water, sodium concentration in the haemolymph increased for the first 2 days and decreased to the seawater-adapted level within 4 days. When transferred from sea water to fresh water, the sodium concentration decreased markedly for the first 2 days and attained the freshwater adapted level after 4 days. When the larvae in sea water were ligated near the anus, they died within 3 days, showing an increased sodium level in the haemolymph. The larvae ligated at the neck lost considerable body weight and died within 4 days.When the anal portion, a terminal portion of the hindgut, was catheterized, the larvae maintained in sea water showed an increase in haemolymph sodium. The anal portion epithelium of the larvae adapted to 100 and 150% sea water demonstrated a strong positive reaction to the histochemical assay for chloride ions, whereas the reaction was negative or weakly positive in freshwater adapted larvae. In the larvae with the anal papillae ligated, a slight increase in haemolymph sodium occurred while in sea water. The anal papillae were weakly positive to chloride ions. Unlike salt-water mosquito larvae of the other species, in which the rectum is considered to be involved in hyperosmotic urine production and the anal papillae appear to be the extrarenal organ, the anal portion inA. togoi larvae seems to play an important role in excretion of excess ions when placed in hyperosmotic media.     

Seasonal changes in the hypo-osmoregulatory ability of brook charr: the role of environmental factors

In winter, brook charr Salvelinus fontinalis demonstrated poor osmoregulatory ability. Saltwater (SW) transfer resulted in sharp increases in plasma osmolality and chloride levels, a decrease in white muscle water content and high mortality. A clear improvement of the hypo-osmoregulatory mechanism e.ciency was observed in spring; by May or June, fish achieved full acclimation within 1 week of SW transfer. In early fall, the time needed to restore osmotic and ionic balance increased once again. A high level of stress was detected only in winter and fall, as indicated by plasma cortisol and glucose concentrations. Great improvement in osmotic and ionic regulation was observed when brook charr were transferred to 80 C SW in February and April while impaired SW adaptability was recorded in individuals transferred to 2·50° C SW in June and August. Temperature itself was more important than a thermal gradient between fresh and salt water. Stress indicators substantiate this temperature-dependent improvement of osmotic and ionic regulation in brook charr. The results support the hypothesis of a minor contribution of cortisol to hypo-osmotic regulation in brook charr, as the cortisol response always seemed to be associated with stress in the experiments.     

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.     

The African lungfish, Protopterus dolloi, detoxifies ammonia to urea during environmental ammonia exposure

The African lungfish, Protopterus dolloi, was able to maintain a low level of blood plasma ammonia during exposure to high concentrations of environmental ammonia. After 6 d of exposure to 30 or 100 mM NH(4)Cl, the total ammonia concentrations in the blood plasma were 0.288 and 0.289 mM, respectively, which were only 1.7-fold greater than the control value of 0.163 mM. In addition, accumulation of ammonia occurred only in the muscle, but not in the liver. This was achieved in part through urea synthesis, as reflected by significant increases in urea contents in the muscle, liver, and plasma of the experimental animals. In contrast with plasma ammonia, the plasma urea concentrations of specimens exposed to 30 or 100 mM NH(4)Cl for 6 d increased 15.4-fold and 18.8-fold, respectively. Taken together, these results suggest that P. dolloi upregulated the rate of urea synthesis to detoxify ammonia during environmental ammonia exposure and that the increased rate of urea synthesis was fast enough to compensate for the rate of endogenous ammonia production plus the net influx of exogenous ammonia in these experimental animals. Simultaneously, there were increases in the rates of urea excretion in the experimental animals between day 2 and day 6 of environmental ammonia exposure. Interestingly, the rates of urea excretion in specimens exposed to 100 mM NH(4)Cl were lower than those exposed to 30 mM NH(4)Cl, despite the presumably greater load of ammonia to be detoxified to urea in the former situation. It would appear that P. dolloi was regulating the rate of urea excretion during ammonia exposure to retain urea, which might have some physiological functions under environmental stresses yet to be determined. There were decreases in the contents of glutamate, glutamine, and total free amino acids in the liver of the experimental animals, which indirectly suggest that a reduction in the rate of proteolysis and/or amino acid catabolism would have occurred that might lead to a decrease in ammonia production. Our results suggest that, unlike marine elasmobranchs and coelacanths, which synthesize and retain urea for osmoregulatory purposes, the ureogenic P. dolloi was adapted to synthesizing and excreting urea for the purpose of ammonia detoxification.     

Effects of environmental water salinity on blood acid-base status in juvenile turbot (Scophthalmus maximus L.)

The time courses of extracellular ionic and acid-base adjustments were studied in juvenile turbot (Scophthalmus maximus) following a decrease of water salinity, either abruptly from 32 to 10%. or after a first step (4 weeks) in 19%. salinity followed by a direct transfer to 10%. brackish water (BW). Net exchanges of acid-base equivalents with the external water were also determined after transfer from 32%. SW to 10%. BW. Direct transfer from seawater (SW) to 10%. BW induced a transient decrease in plasma osmolarity, plasma sodium and chloride concentrations, associated with a marked and transient metabolic alkalosis in the blood. A significant net outflux of acidic equivalents was also measured only during the first day in BW. Four weeks preadaptation in 19%. BW reduced the intensity of the osmotic disturbances elicited by a subsequent abrupt transfer to 10%. BW. These ionic readjustments were also coupled with minimal acid-base changes, of lesser magnitude than those described after directly from SW to 10%. BW.     

Effects of lactate concentration on hybridoma culture in lactate-controlled fed-batch operation

To investigate the effects of lactate on cell growth and antibody production, a new method of maintaining the lactate concentration constant in a fed-batch culture is described. When the pH was initially adjusted by sodium hydroxide, the specific growth rate decreased and specific death rate increased with an increase of lactate concentration. To investigate whether the inhibition was due to the lactate concentration itself or to the osmotic pressure, the effect of the osmotic pressure adjusted by sodium chloride was compared with that of sodium lactate. When the osmotic pressure was adjusted to same condition as that of sodium lactate using sodium chloride, the specific growth data showed the same degree of growth inhibition. It was thus evident that the inhibition to cell growth was mainly due to osmotic pressure while lactate production from glucose was found to be inhibited by the lactate itself compared with sodium chloride. The specific antibody production rate had a maximum value within a certain range of lactate concentration. Moreover, specific antibody production rate had a unified relationship with the kinetic parameter mu, in spite of the different causes of inhibition by lithium lactate and sodium lactate. A certain "trade-off" relationship between growth and antibody production existed at higher growth rates.     

Calcium-sensitive cells of the pars intermedia and osmotic balance in the eel

Summary Freshwater eels were adapted to calcium-free sea water (SW) or 1/3 Ca-free SW. Survival was generally poor in Ca-free SW, although three eels were still in good condition after 19–30 days; survival in 1/3 Ca-free SW was excellent. Osmotic disturbances (increase of plasma osmolarity and sodium levels), which initially occur in Ca-free SW, were no longer detectable after 19–30 days, or in eels in 1/3 Ca-free SW after one month. Plasma calcium sharply decreases initially; it is less depressed after 19–30 days and in 1/3 Ca-free SW. Alterations in the mucus production may be involved in the osmotic changes. Under these conditions no clear stimulation of the calcium-sensitive (Ca-s) cells of the pars intermedia was registered, but in Ca-free SW (1/3 or full strength) the inhibitory effect normally observed in SW does not occur. In a hyperosmotic environment, other ion(s), possibly magnesium, may reduce the response of the Ca-s cells to a lack of environmental calcium.     

Effects of environmental dilution on body fluid regulation in the yellow stingray, Urolophus jamaicensis

Adult yellow stingrays (Urolophus jamaicensis), collected off the southeast Florida coast, were maintained in filtered and re-circulated synthetic sea water (33 per thousand) for 5-13 days at 30 degrees C. Animals exposed to 82%, 74% and 66% SW in gradual steps exhibited rapid and significant weight gains followed by recovery to pre-dilution levels in 2-6 days. Acclimated animals at each salinity [100% (N=12), 82% (7), 74% (4) and 66% SW (3)] were anesthetized (MS222) and bled from the caudal vein. In 100% SW, stingray plasma was slightly hypo-osmotic to the external medium. Plasma osmolality decreased with stepwise dilutions, but became increasingly hyperosmotic to the bathing media. Plasma [Na] and [Cl] each decreased by approximately 13%, 23% and 16%, respectively, in 82%, 74% and 66% SW. Plasma [urea] decreased by 21%, 25% and 59%, respectively. Changes in plasma [K] and [Ca] were minor. Mean corpuscular [Hb] measurements suggest that stingray red cells swelled less at each dilution than predicted for a passive erythrocyte osmometer. RBC [K] decreased by 12%, 36% and 29%, respectively, in 82%, 74% and 66% SW. Quantitatively, the other measured electrolytes (Cl, Na and Ca) changed by lesser amounts. Results suggest that for mild and moderate dilutions (82% and 74% SW), yellow stingrays release both ions and urea from intracellular and extracellular compartments. With further dilution (66% SW), the elasmobranchs retain electrolytes at the expense of urea.     

Osmotic and metabolic responses to dehydration and urea-loading in a dormant,terrestrially hibernating frog

Physiological responses to dehydration in amphibians are reasonably well documented, although little work has addressed this problem in hibernating animals. We investigated osmotic and metabolic responses to experimental manipulation of hydration state in the wood frog (Rana sylvatica), a terrestrial hibernator that encounters low environmental water potential during autumn and winter. In winter-conditioned frogs, plasma osmolality varied inversely with body water content (range 69–79%, fresh mass) primarily due to increases in sodium and chloride concentrations, as well as accumulation of glucose and urea. Decreased hydration was accompanied by a marked reduction in the resting rate of oxygen consumption, which was inversely correlated with plasma osmolality and urea concentration. In a separate experiment, resting rates of oxygen consumption in fully hydrated frogs receiving injections of saline or saline containing urea did not differ initially; however, upon dehydration, metabolic rates decreased sooner in the urea-loaded frogs than in control frogs. Our findings suggest an important role for urea, acting in concert with dehydration, in the metabolic regulation and energy conservation of hibernating R. sylvatica.