Respiratory and osmoregulatory responses of the hermit crab, Clibanarius vittatus (Bosc), to salinity changes

Intertidal hermit crabs were stepwise acclimated to 10, 20, and 30‰ salinity (S) and 21 ± 1 °C. Hemolymph osmolality, sodium, chloride, and magnesium were isosmotic (isoionic) to ambient sea water at 30‰ and hyperosmotic (hyperionic) at 20 and 10‰ S, while hemolymph potassium was significantly hyperionic in all acclimation salinities. Total body water did not differ significantly at any acclimation salinity. Oxygen uptake rates were higher in summer-than winter-adapted crabs. No salinity effect on oxygen consumption occurred in winter-adapted individuals. Summer-adapted, 30‰ acclimated crabs had a significantly lower oxygen consumption rate than those acclimated 10 and 20‰ S. Crabs exposed to 30 10 30‰ and 10 30 10‰ semidiurnal (12 h) and diurnal (24.8 h) fluctuating salinity regimes showed variable osmoregulatory and respiratory responses. Hemolymph osmolality followed the osmolality of the fluctuating ambient sea water in all cases, but was regulated hyperosmotically. Hemolymph sodium, chloride, and magnesium concentrations were similar to hemolymph osmolality changes. Sodium levels fluctuated the least. Hemolymph potassium was regulated hyperionically during all fluctuation patters, but corresponded to sea water potassium only under diurnal conditions. The osmoregulatory ability of Clibanarius vittatus (Bosc) resembles that reported for several euryhaline brachyuran species. The time course of normalized oxygen consumption rate changed inversely with salinity under semidiurnal and diurnal 10 30 10‰ S fluctuations. Patterns of 30 10 30‰ S cycles had no effect on oxygen consumption rate time course changes. The average hourly oxygen consumption rates during both semidiurnal fluctuations were significantly lower than respective control rates, but no statistical difference was observed under diurnal conditions.     

Effects of Low Salinity on Adult Behavior and Larval Performance in the Intertidal Gastropod Crepipatella peruviana (Calyptraeidae)

Shallow-water coastal areas suffer frequent reductions in salinity due to heavy rains, potentially stressing the organisms found there, particularly the early stages of development (including pelagic larvae). Individual adults and newly hatched larvae of the gastropod Crepipatella peruviana were exposed to different levels of salinity stress (32(control), 25, 20 or 15), to quantify the immediate effects of exposure to low salinities on adult and larval behavior and on the physiological performance of the larvae. For adults we recorded the threshold salinity that initiates brood chamber isolation. For larvae, we measured the impact of reduced salinity on velar surface area, velum activity, swimming velocity, clearance rate (CR), oxygen consumption (OCR), and mortality (LC50); we also documented the impact of salinity discontinuities on the vertical distribution of veliger larvae in the water column. The results indicate that adults will completely isolate themselves from the external environment by clamping firmly against the substrate at salinities ≤24. Moreover, the newly hatched larvae showed increased mortality at lower salinities, while survivors showed decreased velum activity, decreased exposed velum surface area, and decreased mean swimming velocity. The clearance rates and oxygen consumption rates of stressed larvae were significantly lower than those of control individuals. Finally, salinity discontinuities affected the vertical distribution of larvae in the water column. Although adults can protect their embryos from low salinity stress until hatching, salinities <24 clearly affect survival, physiology and behavior in early larval life, which will substantially affect the fitness of the species under declining ambient salinities.     

The combined effects of temperature,salinity, and declining oxygen tension on oxygen consumption in the marine pulmonate Amphibola crenata (Gmelin, 1791)

Oxygen consumption of Amphibola crenata (Gmelin) was measured in various salinity-temperature combinations (< 0.1‰ to 41‰ salinity and 5 to 30°C) in air, and following exposure to declining oxygen tensions. In all experimental conditions, respiration varied with the 0.44 power of the body weight (sd = 0.14). The aquatic rate was consistently higher than the aerial rate of oxygen consumption, although at 30 °C the two rates were similar. Oxygen consumption increased with temperature up to 25 °C in all salinities; the lowest values were recorded at temperatures below 10 °C and at 30 °C in the most dilute medium. At all exposure temperatures, the oxygen consumption of Amphibola decreased regularly with salinity down to 0.1 ‰, and following exposure to concentrated sea water (41‰). Salinity had the least effect at 15 °C which was the acclimation temperature. In general, all of the temperature coefficients (Q₁₀ values) were low, < 1.65. However, Q₁₀ values above 2.8 were recorded at a salinity of 17.8‰ between 10 and 15 °C. Oxygen consumption of all size classes of Amphibola was more temperature dependent in air than in water and small individuals show a greater difference between their aerial and aquatic rates than larger snails. The rates of oxygen consumption in declining oxygen tensions were expressed as fractions of the rates in air saturated sea water at each experimental salinity-temperature combination. The quadratic coefficient B₂ becomes increasingly more negative with both decreasing salinity and temperatures up to 20 °C. At higher temperatures (25 and 30 °C) the response is reversed such that O₂ uptake in snails becomes increasingly independent of declining oxygen tensions at higher salinities. On exposure to a salinity of 4‰, Amphibola showed no systematic response to declining oxygen tension with respect to temperature. The ability of Amphibola to maintain its rate of oxygen consumption in a wide range of environmental conditions is discussed in relation to its potential for invading terrestrial habitats and its widespread distribution on New Zealand's intertidal mudflats.     

Effects of salinity on the respiration and heart rate of the common mussel, Mytilus edulis L., and the black chiton, Katherina tunicata (Wood)

The rate of oxygen consumption of stepwise acclimated Mytilus edulis L. increased linearly from 30 to 10‰ salinity (S) while that of Katherina tunicata (Wood) was not significantly different between 10 and 30‰ S. Heart rate was 21–22 and 17–18 beats m^?1 in Mytilus edulis and Katherina tunicata, respectively, and no difference was found in the heart rate of either species acclimated stepwise to 10, 20 or 30‰ S. The average oxygen consumption rate of Mytilus edulis exposed to 12 h, 30-10-30 and 10-30-10‰ S cycles of fluctuating salinity was significantly lower than the respective control rate: there was a similar response during the 30-10-30‰ S cycle in Katherina tunicata. The respiration rate of Mytilus edulis and Katherina tunicata declined as salinity deviated from the control salinity and increased as salinity returned to the control salinity. The rate of oxygen consumption by K. tunicata varied directly with the ambient salinity during the 10-30-10‰ S cycle. The average heart rate of Mytilus edulis was significantly lower during cyclic changes in salinity than at the respective control salinities; a similar relationship existed for Katherina tunicata during the 10-30-10‰ S cycle. Heart rate of Mytilus edulis varied in a parallel manner with oxygen consumption during both cycles. Katherina tunicata heart rate was relatively constant and could not be fitted to a regression line during the 10-30-10‰ S cycle. The normalized heart rate increased to 113% of control at 10‰ S of the 30-10-30‰ S cycle and returned to the control rate by 12 h. The oxygen consumption and heart rate of these two species are not directly coupled to regulation of water volume because different responses are observed with respect to salinity although there is poor water volume regulation in both species.     

Behaviour of Scrobicularia plana (Da Costa) in water of various salinities

Valve movements, heart and pumping rates have been recorded from Scrobicularia plana (Da Costa) when transferred stepwise from natural sea water to sea water of S = 30% (termed 100%) and then to 80, 60, 40, and 20% sea water, and after direct transfer from 100 to 20% Scrobicularia exhibits short periods of pumping alternating with short ventilatory pauses down to 40% sea water. These are termed activity cycles and contrast with periods of complete quiescence each ≈ 4–12 h long. The heart rate of animals in these salinities is fairly constant during activity cycles. When transferred to 20% sea water, either directly or gradually, short pumping phases alternate with long ventilatory pauses during the initial 72 h. At the same time the heart rate shows wide variation during activity cycles.On transference from 100 to 60% sea water, the valves opened and valve activity pattern returned to normal within 30 min and within 6 h of transfer from 60 into 40% SW. Transfer to 20%, whether stepwise or directly, resulted in valves remaining closed initially for ≈ 10 or 34 h, respectively before opening gradually to expose the mantle margin. The valve adductions were then of reduced frequency and extent.     

Osmoregulation of the Nile crocodile, Crocodylus niloticus, in Lake St. Lucia, Kwazulu/Natal, South Africa

Nile crocodiles of three age classes, hatched in captivity and reared in fresh water, when exposed acutely to water of 17 and 35 ppt NaCl, suffered marked dehydration, were lethargic, ceased to feed and lost mass. When exposed to gradually increasing salinities (3-35 ppt), with a short acclimation period at each salinity, crocodiles survived, continued to feed and increased in mass and size. All age classes had a relatively constant plasma osmolality across the salinity spectrum. Cloacal urine osmolality varied throughout the acclimation experiment, but did not increase with increasing salinity. No significant increase was found in plasma concentrations of any of the osmolytes. There was a trend of decreasing cloacal urine [Na(+)] and [Cl(-)] and increasing cloacal urine [K(+)] with increased salinity, indicating that urine was not an important route for Na(+) and Cl(-) excretion. Crocodiles exposed to saline conditions maintained relatively constant plasma uric acid concentrations, but urinary uric acid concentrations increased markedly with increasing salinities. This suggests that uric acid is the main constituent of nitrogenous waste excretion in saline exposed Nile crocodiles. As in Crocodylus porosus, C.niloticus has the physiological ability to survive and thrive in periodically hyper-osmotic environments. However, its euryhalinity is restricted, in that acute exposure to sea water leads to dehydration, but with an acclimation period at lower salinities, it survives and thrives in sea water.     

Hypoosmotic volume regulation in the sea anemone metridium senile

1. The anemone Metridium senile survives salinities from seawater (950 mOsm) to 55% SW (520 mOsm) for at least two weeks. Animals exposed to 40% SW (380 mOsm) die within three days.2. The tissue amino acid content of M. senile acclimated to 950 mOsm, 807 mOsm, 665 mOsm and 520 mOsm for two weeks is respectively, 444, 382, 331 and 251 μmol/g dry wt. A decrease in the concentration of taurine accounts for nearly all of the decrease in the free amino acid pool.3. Tissue hydration increases in M. senile acclimated to dilute seawater, but the increase was not proportional to the change in ambient salinity, indicating that the anemones partially regulate volume in dilute media.4. Mathematical analyses of changes in tissue hydration as a function of ambient salinity in M. senile, Haliplanella lineata, and Diadumene leucolena suggest that the effectiveness of volume regulation increases in individuals of these species acclimated to progressively more dilute media. The volume regulatory capability of Bunodosoma cavernata does not change in dilute media.     

Behavioral influences on the physiological responses of Cancer gracilis, the graceful crab, during hyposaline exposure

The relationship between the behavioral and physiological responses to hyposaline exposure was investigated in Cancer gracilis, the graceful crab. The status of C. gracilis as an osmoconformer was confirmed. Survival decreased with salinity: the LT(50) in 50% seawater (a practical salinity of 16, or 16 per thousand) was 31.5 +/- 22.7 h and in 25% seawater (a salinity of 8) was 8.0 +/- 0.7 h. When exposed to a salinity gradient, most crabs moved towards the highest salinity. However, in the salinity range of 55% to 65% seawater, they became quiescent. This "closure response" was also evident at low salinities: the mouthparts were tightly closed and animals remained motionless for 2 to 2.5 h. During closure, crabs were able to maintain the salinity of water within the branchial chambers at a level that was about 30% higher than that of the surrounding medium. The closure response was closely linked to a short-term decrease in oxygen uptake. During closure, oxygen within the branchial chamber was rapidly depleted, with oxygen uptake returning to pretreatment levels upon the resumption of activity. In addition to the short-term decrease in oxygen uptake, there was a longer-term bradycardia, which may serve to further reduce diffusive ion loss across the gills. By exhibiting a closure response during acute hyposaline exposure and an avoidance reaction during prolonged or severe hyposaline exposure, C. gracilis is able to use behavior to exploit areas prone to frequent episodes of low salinity.     

The influence of salinity on water balance in 0-group flounders, Platichthys flesus (L)

The water balance of 0-group flounders was investigated in a range of static [0–100% sea water (SW), 100%≡ 34%‰] and cycling salinities (2–98% SW, 12 h 25 min period). The permeability coefficients of these juvenile fish were found to be higher than those quoted for adults. The permeability of juveniles in fixed salinities decreased with increased salinity whereas animals in the tidally cycling regime showed permeability changes that were directly proportional to the ambient salinity. However, comparison of the two groups showed that animals in a cycling salinity regime were less permeable to water than animals acclimated to fixed salinities. Drinking and urine production rates fluctuated within a tidal salinity regime, and 0-group flounders were found to modify their water permeability, urine production and drinking rates simultaneously, so maintaining their blood osmotic concentration and total water content within narrow limits throughout the range of salinities.
The relevance of measurements made in fixed and tidally cycling salinities to water regulation under natural estuarine conditions is considered.     

O2 consumption and acute salinity exposure in the freshwater shrimp Macrobrachium olfersii (Wiegmann) (Crustacea:Decapoda): whole animal and tissue respiration

The time course of O₂ consumption after acute salinity exposure (1, 3, 6, 12, and 24 h to 0, 7, 14, 21, 28, and 35 S) was examined in isolated supraesophageal ganglia, gills, and intact Macrobrachium olfersii (Wiegmann), a hyperosmoregulating freshwater palaemonid shrimp, to establish patterns of metabolic adjustment during salinity adaptation. In whole shrimps, O₂ uptake rates decline with salinity increase to 21 S, subsequently increasing with further salinity increase. The rates increase to maxima after 6–12-h exposure in low salinities, decreasing steadily with time in high salinities. In gill preparations, O₂ consumption rates increase to a maximum in 14 S, then decline; they are maximal after 3–6-h exposure to low salinities and diminish with time in high salinities. In the supraesophageal ganglion, rates of O₂ uptake, always measured in seawater of 18 S, are also maximal when shrimps are exposed to 14 S, subsequently declining or levelling off. Rates decrease with time in shrimps exposed to very low salinities, and are stable in 21 S, reaching maxima after 3–6-h exposure of shrimps to all other media. Both tissues thus exhibit characteristic response patterns of O₂ consumption rate which appear to depend on their functional significance within the context of the whole organism. Such data are interpreted to indicate an interrelationship between O₂ consumption and osmoregulatory capability.     

Ecological physiology of arctic marine invertebrates. Temperature and salinity relationships of the amphipod Onisimus affinis H. J. Hansen

Effects of temperature and salinity upon the survival, locomotion and metabolism of the Arctic marine amphipod Onisimus affinis H. J. Hansen have been investigated. The LD₅₀ for temperature is ≈ 18.7 °C. The metabolic rate-temperature curve shows a distinct plateau of relative temperature insensitivity the position of which varies seasonally to include a lower temperature range in winter than in summer. Similar shifts in the plateau can be induced in the laboratory by acclimating the animals at summer- and winter-like temperatures.Optimal locomotory activity was between 5° and 8 °C and included a combination of swimming and crawling. Above 12 °C the swimming component was increasingly inhibited.Onisimus is euryhaline and appears to be most successful in brackish water habitats. It tolerates elevated salinities better at low temperatures. The metabolic rate varies inversely with salinity during short-term exposures, but, if the animals are pre-adapted to the experimental salinities for 10 days, the metabolic rate is essentially independent of salinity between 10%. and 25%.The significance of these physiological responses in relation to the general ecology of the species is discussed.     

Effects of low salinity on juvenile pink shrimp Farfantepenaeus paulensis (Perez-Farfante 1967, Crustacea)

It is of paramount importance to study salinity tolerance of commercially important crustaceans, such as the pink shrimp Farfantepenaeus paulensis to determine possible mortality causes in the wild and in aquaculture in oligohaline waters. The aim of this study was to determine the lethal salinity concentration (LC50) for juvenile pink shrimp F. paulensis and measure its oxygen consumption and ammonia excretion at different salinity levels. Shrimp of two length classes (49.4?±?4.3 and 78.5?±?5.5?mm) were placed in 10-L containers and exposed to salinity levels of 35, 30, 25, 20, 15, 10 and 5. The experiments were tripled, with seven shrimp in each container. The average lethal concentration (LC50s) for an exposure of 24?h was 13.33 (11.26–15.78) and 10.26 (8.60–12.64), respectively, for the two classes of juveniles. For an exposure of 48?h, LC50s were 12.71 (10.68–15.12) for the larger animals and 9.20 (7.34–11.52) for the smaller ones. There was an inverse relationship between salinity and rates of oxygen consumption and ammonia excretion. The average reduction in specific oxygen consumption in salinities 20, 25 and 30 showed a decrease in metabolic rate of 63, 80 and 82%, respectively, in relation to salinity level 0. The same occurred for the averages of ammonia excretion at salinity levels of 15, 20, 25, 30 and 35, which represented low metabolic rates of 57, 61, 70, 71 and 74% respectably in relation to salinity level 0.     

Effect of salinity on oxygen consumption in fishes: a review

The effect of salinity on resting oxygen uptake was measured in the perch Perca fluviatilis and available information on oxygen uptake in teleost species at a variety of salinities was reviewed. Trans‐epithelial ion transport against a concentration gradient requires energy and exposure to salinities osmotically different from the body fluids therefore imposes an energetic demand that is expected to be lowest in brackish water compared to fresh and sea water. Across species, there is no clear trend between oxygen uptake and salinity, and estimates of cost of osmotic and ionic regulation vary from a few per cent to >30% of standard metabolism.     

Effect of eyestalk ablation on oxygen consumption of callinectes similis exposed to salinity changes

• 1.1. The effect of eyestalk ablation on preadults of Callinectes similis exposed to a constant salinity (30%.) and to simulated tidal changes in salinity (30-11 to 30%.) were measured.
• 2.2. In constant salinity, crabs showed a persistent respiratory rhythm, with a maximum oxygen consumption during the day. Under these conditions, ablation significantly increased the respiratory rate but not the rhythm.
• 3.3. In variable salinities, the highest respiratory rates occurred in salinities of 11 and 16%. during the night. In these crabs, ablation of eyestalks and subsequent injection of eyestalk extracts did not alter the respiration rate rhythm.
• 4.4. The circadian rhythm is controlled by the periodicity of environmental changes instead of the influence of eyestalk hormones.
• 5.5. Regulation of metabolism in C. similis associated with osmoregulation involves other neurosecretory organs.

     

Effects of salinity and cyclic temperature on survival of two sympatric species of grass shrimp (Palaemonetes), and their relationship to natural distributions

Survival and respiration of the grass shrimp Palaemonetes pugio Holthuis and P. vulgaris (Say) from the Newport River estuary were measured after exposure to cyclic and constant winter temperatures, to rapid decreases in temperature, and to various temperature-salinity combinations. Both species were subjected to nine temperature-salinity combinations including temperature regimes of cyclic 7–13°C, constant 7° and 10°C, and salinities of 5, 20, and 35%.. Based on the laboratory and field results, the differences in physiological tolerance to winter temperatures and salinities were examined in relation to habitat partitioning by these sympatric species.Survival after continuous exposure to cyclic temperature regimes at medium to high salinities was similar to that observed for comparable constant temperatures; however, at low salinities mortality was significantly lower under the cyclic regime than under either constant regime. This suggests that cyclic temperatures may be detrimental in combination with some other stress. A rapid, transient decrease in temperature from either 7° or 10°C to 2°C had no measurable effect on survival or rate of oxygen consumption at any temperature-salinity acclimation. Neither salinity (except in areas intermittently subjected to salinities below 3 %.) nor winter temperatures appear to affect habitat partitioning in grass shrimp.     

Effects of salinity and temperature on oxygen consumption in a freshwater population of Palaemonetes antennarius (Crustacea,decapoda)

• 1.1. After step-like increases in salinity the shrimps exhibit the smallest increase in oxygen consumption in the lower salinity range. At higher salinities the shrimps show longer recovery times and greater increases in the metabolic rate after salinity shock.
• 2.2. In steady-state experiments, the shrimps display the lowest oxygen consumption rates near the isosmotic point. The lowest metabolic rates occur at salinities of 3‰ and 10‰ At salinities of 20‰ and above the rate of metabolism increases by 20–30%.
• 3.3. The calculated osmoregulatory work for animals in fresh water amounts to only 2.7% of routine metabolism and drops to 1.1% for shrimps in 3‰ and 0.7% in 5‰ salinity.
• 4.4. Locomotory activity in the form of position change was not responsible for the increased oxygen consumption of the animals after salinity shocks. A “tentative swimming activity” by fast and frequent beating of the pleopods without position change may be an important factor in the increase of metabolic rates.
• 5.5. In its temperature response, the brackish water population has a higher metabolic rate than the freshwater one. Between 5 and 35°C Q ₁₀-values range from 4.01 to 1.37.

     

Respiration in air and water of three mangrove snails

The oxygen consumption of three species of Malaysian mangrove gastropods was measured in air and sea water at the temperatures commonly recorded in the mangrove. The experiments in air were carried out after the animals had regained fluid lost from the mantle due to handling. Fluid loss can have considerable effects on rate of oxygen consumption. Nerita arliculata (Gould) was found throughout the mangrove and experiences from 50 to 92% aerial exposure. It has a gill and a ratio of aerial to aquatic respiration rates of 2.7 at 28°C. 50% of the animals can survive underwater for 72 h at 28°C. The other two species, Cerithidea obtusa (Lamarck) and Cassidula aurisfelis (Brugière) experience over 95% aerial exposure, have their mantle cavities modified as lungs and have air : water respiration rate ratios of 5.5 and 6.0, respectively. 50% can survive from 48 to 36 h underwater at 28°C. Acclimated animals have Q₁₀'s of about 1.6 in air and 1.4 in water. The respiratory physiology of the snails is compared with that of rocky shore species.     

Microgrowth patterns in the shell of the Malaysian cockle Anadara granosa (L.) and their use in age determination

Marked specimens of the Malaysian cockle Anadara granosa (L.) were placed in buried cages (filled with sediment from the surrounding substratum) which were located in intertidal and subtidal environments in an area between Penang Island and mainland West Malaysia. In the majority of cockle shells examined the number of growth bands deposited was close to the number of tidal periods. Bands in the shells of intertidal animals were more strongly defined than those in shells continuously immersed. The banding pattern in shells from the subtidal environments showed narrow increments during spring tides alternating with a few wider increments during neap tides. The growth increments between bands correspond to tidal periods so they can be used to record growth rates and provide estimates of the ages of the shells. Samples of cockles collected from four sites in a commercial cockle-culture area off the coast of West Malaysia were aged. Two of the sites were located in an estuary where the cockles were periodically exposed to fluctuating salinities during the intermonsoon period. Cockles from the other two sites were situated away from the estuary and experienced full strength sea water. There was good agreement between the calculated estimates of the ages determined from the tidal bands and their known ages. Shells of cockles from the estuary had characteristically marked banding patterns which may be related to shell deposition during the intermonsoon period when the cockles were exposed to sea water of low salinity. Cockle shells collected outside the estuary did not display these patterns.     

Gradual transfer to sea water of rainbow trout: effects on liver carbohydrate metabolism

The levels of glycogen and glucose, as well as of the activities of several key enzymes of glycogenolysis, glycolysis, gluconeogenesis and the pentose phosphate shunt were assessed in livers of small (97 g) and large (166 g) rainbow trout Oncorhynchus mykiss during gradual transfer to sea water (salinities of 0, 9, 18 and 28%). Plasma glucose and protein levels were also evaluated. Liver carbohydrate metabolism changed during gradual adaptation of rainbow trout to sea water. Glucose increased with salinity in livers of both sizes of animals, as did glycogenolysis and gluconeogenesis, but this latter only in large animals, and glycolytic rates in small animals. The different responses in metabolic parameters between sizes of animals may reflect a higher stress of the small animals and/or a better adaptation of large animals to increased salinities, in a way similar to that previously suggested by other authors after evaluation of non-metabolic parameters. The changes observed at 28% can also be the result of reduced food consumption.     

Effects of salinity on synthesis of DNA,acidic polysaccharide,and ichthyotoxin in Gymnodinium breve

A Florida red tide organism, Gymnodinium breve Davis, an unarmored dinoflagellate, was grown in enriched sea water media at salinities 20–43% and constant illumination. Use of lowest (23%) and highest (43%) salinities resulted in death within 24 hr of inoculation, though good growth was obtained at all intermediate salinities (29–39%), in accord with field observation. Rates of synthesis of DNA, acidic polysaccharide and ichthyotoxin were determined as a function of salinity and growth constant (K₁₀). The relative rate of' synthesis of DNA or polysaccharide increased linearly with growth constant. Mean cell volumes, determined during log-phase growth, showed a positive correlation with doubling time. Hemolytic activity was detected in cell extracts only at high toxin concentrations (0.35–2.05 mg of ichthyotoxin). No significant difference was noted in hemolytic activity of extracts of cells grown in high (34%) or low (26%) salinity. The rate of toxin synthesis showed a linear decrease with the rate of DNA or polysaccharide synthesis.