Water balance and osmoregulation in Stenocara gracilipes,a wax-blooming tenebrionid beetle from the Namib Desert

Dehydration (10 days at 27 degrees C) of the Namib tenebrionid Stenocara gracilipes resulted in a rapid weight loss (17.5%), and a substantial decline in haemolymph volume (72%). Although the lipid content decreased significantly, metabolic water production was insufficient to maintain total body water (TBW). Rehydration (no food) resulted in increases in haemolymph volume, body weight (sub-normal), and TBW to normality. Haemolymph osmolality, sodium, potassium, chloride, amino acids, and sugars (trehalose and glucose), were all subject to osmoregulatory control during both dehydration and rehydration. Major osmolar effectors in this species are sodium, chloride, and amino acids, with most of the contribution to regulation of haemolymph osmolality coming from changes in the levels of these constituents. Changes in amino acid levels are not the result of interchange with soluble protein during dehydration (the possibility exists during extended rehydration, however). Despite faecal losses of sodium being low (8.2% of that removed from the haemolymph during dehydration), sodium concentrations do not return to normal during rehydration. Chloride concentrations increase supra-normally when access to water is allowed, and remain elevated throughout the rehydration period. Although faecal loss of potassium greatly exceeded the amount removed from the haemolymph (by approximately 1.8 times), haemolymph potassium levels were strongly regulated during rehydration. S. gracilipes demonstrates an exquisite capacity to regulate haemolymph osmolality under conditions of both acute water-shortage and -abundance. Together with an efficient water economy (drinking when fog-water is available, and a superb water conservation mechanism in the form of wax-bloom production), this must serve to contribute to long-term survival of this species in an otherwise harsh abode.     

Water balance and osmoregulation in Onymacris plana, a tenebrionid beetle from the Namib desert

Onymacris plana, a tenebrionid beetle from the sand dunes of the Namib desert, lost weight very slowly during 12 days of dehydration at 26°C. Measurement of total lipid showed a gradual decline, the metabolic water produced being sufficient to maintain a constant water content. At the same time the haemolymph volume decreased by 66%. When given water the dehydrated beetles drank rapidly and their weight and haemolymph volume were restored to normal. Haemolymph osmolarity was closely regulated despite the changes in volume. Haemolymph potassium was also well regulated, but sodium was lost from the haemolymph during a cycle of dehydration and rehydration, even though sodium losses in the faeces were small. Water balance in Onymacris depends on efficient conservation of water in periods of drought and on water uptake by drinking during the coastal fogs of the Namib.     

Effects of starvation and dehydration on ionic balance in Schistocerca gregaria

This study describes the effects that prolonged dehydration has on ionic balance in Schistocerca gregaria. When adult locusts are dehydrated for 7 days the body weight reduces by 10–20% and the haemolymph volume by 35–50%, but haemolymph concentrations of Na⁺, K⁺ and Cl⁻ change only slightly. On dehydration Na⁺ and Cl⁻ are removed from the haemolymph; 25% of the removed ions is excreted and 75% is evenly distributed in the body of the locust. The amount of potassium excreted always exceeds that removed from the haemolymph. Mature adults control more effectively than young ones the haemolymph ionic composition during dehydration, but young adults show a smaller reduction in haemolymph volume. In the normal state of hydration, 76% of the total body Na⁺ and 56% of the total body Cl⁻ is present in the haemolymph. These fall to 62 and 42% respectively on dehydration and increase to 77 and 50% on rehydration.     

The effects of desiccation and rehydration on the lone star tick

This study describes the effects of desiccation and rehydration on the water content, haemolymph volume (per cent), osmolarity, and concentrations of Na, K, Mg, and Ca in the haemolymph of the lone star tick, Amblyomma americanum.The water content percentages of ‘severely desiccated’, ‘moderately’ and ‘fully hydrated’ ticks were 46·0, 52·8, and 60·3 per cent respectively. The lowest and highest of these were near the minimum and maximum possible.The haemolymph volume (per cent) of ‘severely desiccated’ ticks was regulated near the level of ‘moderately hydrated’ ticks despite significant decreases in total body water content and increases in osmolarity and concentration of sodium. Conversely, the change from ‘severely desiccated’ to ‘moderately hydrated’ ticks can be viewed as causing an increase in total body water, decrease in blood osmolarity and sodium, but little change in haemolymph volume (per cent).Most of the water taken up by ‘moderately hydrated’ ticks (while becoming ‘fully hydrated’) was added to the haemolymph. At the same time, there was little change in the blood osmolarity or haemolymph concentration of sodium. Conversely, the change of ‘fully’ to ‘moderately hydrated’ ticks was marked by a substantial loss of haemolymph volume (per cent) but little change in osmolarity and concentration of sodium.The concentration of potassium was regulated over the full range of desiccating and hydrating conditions. The lone star tick appeared less able to regulate its haemolymph concentrations of Ca and Mg; both fluctuated at the same rate, but inversely as the haemolymph volume (per cent).It appears that a carefully controlled movement of solutes (Na the predominant cation) between haemolymph and non-haemolymph tissue is intimately linked with haemolymph volume regulation and movement of water into the haemolymph during hydration.     

Tyrosine and phenylalanine concentrations in haemolymph and tissues of the American cockroach, Periplaneta americana, during metamorphosis

Phenylalanine and tyrosine concentrations were measured in the haemolymph, fat body, and abdominal integument of the American cockroach, Periplaneta americana, during the pre- and post-ecdysial periods of cuticle formation and sclerotization.Gas-liquid chromatography of trimethylsilyl derivatives of phenylalanine, tyrosine, and their metabolites provided a very sensitive and rapid method for determining those amino acids in small haemolymph and tissue samples.Haemolymph tyrosine increased in two stages: initially near apolysis and 16 to 25 hr pre-ecdysis, reaching its highest concentration at ecdysis (3·5 μg tyrosine/mg haemolymph). During that time, total haemolymph tyrosine increased by approximately 700 μg/insect. Fat body and abdominal integument began to accumulate tyrosine near apolysis. Fat body tyrosine peaked between ecdysis and 3·3 hr post-ecdysis whereas abdominal integument tyrosine peaked at ecdysis. Maximum concentrations were 6·0 μg and 4·1 μg tyrosine/mg wet wt. of tissue, respectively. Between ecdysis and 24 hr post-ecdysis, the period of maximum sclerotization, total tyrosine in haemolymph and fat body decreased by approximately 600 μg and 420 μg/insect, respectively. Phenylalanine concentrations did not change significantly in the haemolymph, fat body, or abdominal integument during the pre- and post-ecdysial periods.The cockroach apparently does not store free phenylalanine or tyrosine in the fat body during larval development as compared to tyrosine storage in some Diptera. The rapid increase of haemolymph, fat body, and integument tyrosine just prior to ecdysis suggests another form of storage for this important amino acid.     

The role of the gills in seawater adaptation inAnguilla dieffenbachii

Summary The mean serum sodium, chloride and potassium concentrations and serum osmotic pressure of freshwaterA. dieffenbachii are 140.4 mmol 1^–1,114.0 mmol 1^–1, 6.66 mmol 1^–1 and 307.7 mOsmol respectively. Gill tissue from freshwater specimens has a water content of 4234 mg g dry wt^–1 (80.9% wet wt), a chloride space of 1852 mg water g dry wt^–1 (35.2% wet wt) and an intracellular volume of 2449 mg water g dry wt (46.0% wet wt). Estimates of the intracellular sodium and potassium concentrations for the gill tissue of freshwater eels gave values of 28.9 mmol kg intracellular water^–1 and 126.5 mmol kg intracellular water^–1 respectively. On transfer of the fish to sea water serum concentrations of sodium and chloride and serum osmotic pressure show rapid initial increases followed by a more gradual decline eventually stabilising at new levels some 100 hours after transfer (Fig. 1). The serum sodium, chloride and potassium concentrations and serum osmotic pressure of seawater-adaptedA. dieffenbaehii are 162.8 mmol 1^–1, 151.0 mmol 1^–1, 6.70 mmol 1^–1 and 376.9 mOsmol respectively.On transfer to sea water the water content and chloride space of the gill tissue is reduced and the intracellular volume is initially decreased but is rapidly restored to its original value (Fig. 2, 3). At the same time intracellular sodium and potassium concentrations are increased but the latter is fairly rapidly restored to pre-transfer levels (Fig. 4).The changes in intracellular potassium concentration can be explained largely by the changes in intracellular volume but intracellular sodium concentrations remain high on transfer because of the increased serum sodium concentration. The initial increases in serum concentrations on transfer to sea water are caused partly by the removal of water and partly by the addition of sodium and chloride ions to the internal body fluids.     

X-ray microanalysis of cockroach fat body in relation to ion and water regulation

X-Ray microanalysis of fat body from the cockroach (Periplaneta americana) showed that the crystals in the urate cells sequestered potassium and sodium during water deprivation. Postassium sequestration appeared to occur by the formation of additional crystals whereas sodium appeared to be taken up by pre-existing crystals of the “amorphous granule” type. Urate cells increased in cytoplasmic density i.e. mass per unit volume as a result of water deprivation and lost potassium from the cytoplasm but not from the nucleus. Trophocyte cells also showed an increase in density and a loss of potassium from the cytoplasm. The nucleus in trophocyte cells however, did not increase in density and showed an increase in chlorine concentration. A previously undescribed type of crystal in the trophocyte cell cytoplasm also increased markedly in chlorine concentration. The mycetocyte cells did not appear to increase in density but chlorine concentration was elevated in symbionts, cytoplasm and nucleus. An increase in density is considered to be a consequence of a reduction in volume due to water loss. It is confirmed that the fat body plays a major role in the maintenance of ion balance during water deprivation.     

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.     

Ionic and osmotic regulation of the haemolymph of the dragonfly, Libellula quadrimaculata (Odonata : Libellulidae)

Larvae of the widespread dragonfly, Libellula quadrimaculata, were adapted to a series of salt solutions, and the osmotic pressure, and sodium, potassium and chloride concentrations in the haemolymph measured. The regulation of potassium is extremely efficient over the range 0–50 m-mole/l. external concentration. Above this, larvae die. Sodium and chloride are regulated to a lesser extent, the larvae being able to withstand considerable changes in the concentration of these ions in the haemolymph. However, at higher external concentrations, the haemolymph concentration of these ions is maintained below that of the external medium. The osmotic pressure is regulated in parallel with sodium concentration over most of the range tested. However, in higher salinities, the osmotic pressure of the haemolymph does not fall below that of the external medium. This is seen as a strategy to limit the amount of drinking in saline media. Overall, the osmoregulatory system of L. quadrimaculata resembles that of brackish-water insects, rather than that of the more strictly freshwater dragonflies that have been studied.     

Water relations and haemolymph composition of two intertidal spiders (order araneae)

Water relations and haemolymph composition have been compared in two intertidal spiders which inhabit different zones of a rocky shore in the Cape Peninsula. Evaporative water losses of the mid-shore species, Desis formidabilis (O.P. Cambridge), are much greater than those of Amaurobioides africanus Hewitt, which occurs higher on the shore. Both species avoid desiccating conditions by remaining in silk-lined nests in suitable microclimates, and the nests of Desis formidabilis are submerged for a substantial portion of each tidal cycle. Osmolarities and sodium, potassium, magnesium, and chloride concentrations were measured for the haemolymph of both spiders. The haemolymph chemistry of Amaurobioides africanus resembles that of terrestrial spiders. The haemolymph of Desis formidabilis, however, has an osmolarity of 930 mOsm/l, a sodium concentration of 451 mM/l and a chloride concentration of 466 mM/l. It is suggested that this unusually concentrated haemolymph may represent an adaptation to a diet of marine crustaceans.     

Water balance during vitellogenesis by the American cockroach:distribution of water during the six day cycle.

Haemolymph volume, total body water, and total tissue water fluctuate during the vitellogenic cycle in the American cockroach. The observed patterns indicate that the tissues store water for oöthecal production rather than the haemolymph. The carcass water (which consists of the integument and adhering tissue) cycles in direct contrast to the water ingestion pattern, suggesting that these tissues supply ovariole water incorporation during days 3 and 5. Both fat body and gut release large volumes of water during day 6 to the haemolymph for use in the formation of the oötheca.     

Insect cold tolerance and repair of chill-injury at fluctuating thermal regimes: role of ion homeostasis

Adults of the bug Pyrrhocoris apterus and the beetle Alphitobius diaperinus developed chill-injury slower and survived longer when they were exposed to fluctuating thermal regimes (FTRs, where periods of low temperature were alternated with periods of higher temperature on a daily basis) rather than to constant low temperatures. The extracellular (haemolymph) concentrations of potassium ions increased with significantly higher rates in the insects exposed to constant low temperatures than in those exposed to FTRs. The concentrations of magnesium and sodium ions were maintained relatively constant or decreased slightly in both thermal regimes. The loss of body water and the increase of haemolymph osmolality contributed to, but could not fully explain, the ion concentration changes, which probably resulted also from impairing the function of an active metabolic component (ion pump) at low temperatures. This explanation was supported by observing (in P. apterus) the return toward normal [K+] during the warm "recovery" period of the FTR. Collectively, the paper stresses the importance of considering the temperature fluctuations in the experimental studies on insect cold tolerance and suggests that the positive effect of the FTR on cold tolerance may consist, at least partially, in allowing the primary ion pumping systems to re-establish the ion gradients across cell membranes and epithelia during the recovery periods at a higher temperature.     

Water balance and osmoregulation in a free-ranging tenebrionid beetle, Onymacris unguicularis, of the Namib Desert

Onymacris unguicularis, a fog-basking tenebrionid beetle of the Namib Desert, has mean water influx rates of 49.9 mg H₂O/g.d and mean efflux rates of 41.3 mg H₂O/g.d with mean mass gain being 10.7 mg/g.d. If only steady-state conditions are considered (no mass change), and passive vapour input subtracted, drinking accounts for 50% of water input. Active beetles must drink in order to maintain water balance, while inactive beetles can maintain water balance either eating seeds or by simply metabolizing fat. Little change is observed in ratios of haemolymph and total body water to dry mass when fogs occur, while significant changes in haemolymph osmotic pressure are associated with fog occurrence.For short periods, O. unguicularis can tolerate dehydration with only slight changes in the ratio of total body water and haemolymph to dry body mass and to haemolymph osmotic pressure. For longer periods however, active beetles must have access to fog water for water balance maintenance. This is probably necessary for reproduction.     

Effects of starvation and dehydration on osmotic and ionic balance in Carausius morosus

This study describes the effects that prolonged desiccation has on osmotic and ionic balance in Carausius morosus, an insect with a rather unusual haemolymph composition (it is rich in divalent cations and has a low sodium content). After 7 days at 0% r.h., there is a 50 per cent decrease in water content of the animal. This loss is largely confined to the haemolymph, which declines in volume to about one-seventh of its previous value. In spite of these drastic changes, the osmotic concentration of the haemolymph increases only by about 20 per cent while the concentrations of the main blood ions increase by similar or smaller amounts. So, as with other insects so far examined, Carausius turns out to have very effective control of the osmotic and ionic concentrations of its internal milieu in the face of severe stress upon it.     

The biochemical effects of potassium chloride on the silkworm, （ Bombyx mori L.）

The supplementation with 50, 100 and 150μg/mL potassium chloride to the fifth instar larvae of the silkworm Bombyx mori on fat body glycogen, protein, total lipids and haemolymph protein and trehalose were analyzed. The fat body glycogen and protein and haemolymph protein were increased significantly in all the treated groups; whereas fat body total lipids increased only in 100 and 150μg/mL and haemolymph trehalose increased only in 150μg/mL potassium chloride-treated groups when compared with those of the corresponding parameters of the carrier controls.     

The effects of dietary sodium and potassium on rapid diuresis in the tsetse fly Glossina morsitans

A self-supporting colony of Glossina morsitans can be maintained by feeding flies through artificial membranes but only if a diet of defibrinated pig blood is used. The potassium concentration of pig serum was found to be higher than that of cow or rabbit serum. However, this did not affect the rate of diuresis which was lower after in vitro diets than after feeding on a live host.After feeding in vivo on rabbits, and in vitro on pig blood and cow blood, the concentrations of sodium and potassium in the haemolymph remained constant. After feeding on salt solutions and distilled water, on the other hand, it was found that the more the composition of the meal varied from that of blood serum the lower was the rate of diuresis and the more the concentrations of sodium and potassium in the haemolymph were displaced from their normal values. Large fluctuations in the concentrations of these ions resulted in the paralysis and death of the flies.The results demonstrate that the tsetse fly is able to reabsorb sodium from the primary urine produced by the Malpighian tubules during diuresis and it is suggested that this reabsorption of ions may lead to an increased circulation of water through the excretory system.     

Water balance in Rhodnius prolixus during flight: Quantitative aspects of diuresis and its relation to changes in haemolymph and flight-muscle water

The amount of water voided by male Rhodnius prolixus which were flown to exhaustion varied from 0 to over 10% of the initial live weight. It accounted for nearly all of the body water lost during the flight period. Simultaneous measurements on the loss of haemolymph water and an estimate of the amount of faecal water in the excreta indicated that the source of the voided water was primarily the haemolymph. The total water content of the flight muscles changed very little in insects which flew to exhaustion. It is concluded that, despite the diuresis and loss of water, and the considerable reduction in haemolymph volume, dehydration of the flight muscles of male R. prolixus does not occur during these flight periods, and is not a factor contributing to ‘exhaustion’. The possibility that insufficient haemolymph is a factor limiting the duration of flight is discussed.     

Osmoregulation in Onymacris rugatipennis, a free-ranging tenebrionid beetle from the Namib Desert.

Haemolymph levels of organic and inorganic constituents were investigated in the tenebrionid Onymacris rugatipennis during dehydration and rehydration. The major osmolar effectors are sodium (26%), chloride (24%), amino acids (18%), and sugars (11%); regulation of haemolymph osmotic pressure (OP) during dehydration is effected largely by a reduction in the haemolymph content of these constituents. Changes in amino acid levels are not the result of interchanges with soluble protein. During rehydration, the main contributors to osmoregulation are sodium (26%), chloride (24%), and an increase in haemolymph solute(s) not measured in this study (31%). Of the sodium removed from the haemolymph during dehydration, 21.2% was excreted. Faecal losses of potassium during dehydration far exceeded the amounts removed from the haemolymph; however, haemolymph potassium levels were strongly regulated during rehydration. Regulatory efficiency increases as desiccation proceeds, and is greatest only when this species is severely challenged.     

Effect of elevated temperature on osmotic and ionic regulation in a salt-tolerant caddisfly from Death Valley,California

The effects of temperature (8–10 or 20°C) on regulation of haemolymph osmotic and ionic concentrations were investigated over a range of salinities (0–25‰) in fifth-instar larvae of the Death Valley caddisfly Limnephilus assimilis. At low temperatures, levels of chloride and sodium in the haemolymph are regulated over a wide range of salinities corresponding to the salinities at which larvae occur in nature and at which they can complete development into adults. In contrast, haemolymph osmolality is constant at low salinities (<14‰) but approaches conformity with the medium at higher salinities. High temperature reduces the larva's ability to maintain low chloride concentrations in its haemolymph and also leads to a reduction in haemolymph osmotic pressure; thus, at high temperatures ions account for more of the haemolymph osmotic concentration than at low temperatures. These data suggest that the absence of larvae from thermal pools and from all Death Valley waters in summer can be explained by the effects of high water temperatures on hydromineral regulation.     

The composition of larval-pupal moulting fluid in the tobacco hornworm, Manduca sexta

The concentrations of sodium, potassium, chloride, bicarbonate, total phosphate, labile phosphate, inorganic phosphate, protein, polypeptides, amino acids, trehalose, and glucose, as well as pH and osmotic pressure of larval-pupal moulting fluid and haemolymph were measured in the tobacco hornworm, Manduca sexta, during the secretion and resorption of moulting fluid. Moulting fluid is a mildly alkaline (pH 7.2), iso-osmotic (330 mOsm) potassium bicarbonate salt solution containing within it the sol form of moulting gel. Bicarbonate is the principal anion in moulting fluid. It is only a minor component of haemolymph. Significant differences in the concentrations of potassium, chloride, bicarbonate, non-labile phosphate, labile phosphate, inorganic phosphate, protein, polypeptide, amino acids, trehalose, and glucose indicate that the integumentary epithelium does not act as a semi-permeable membrane, and that free diffusion between moulting fluid and haemolymph can account for only a small fraction of the molecular species movement between these two fluids.