Interaction of Respiration, Ion Regulation, and Acid-Base Balance in the Everyday Life of Aquatic Crustaceans

Extracellular and intracellular acid-base balance is necessaryfor the maintenance of normal metabolic processes. The primarysource of acid is metabolically produced CO₂, and the CO₂/HCO₃^–system is the most significant buffer. The regulation of acid-basebalance is complex, involving the interaction between respiratorygas exchange and ion transport. In aquatic crustaceans respirationis governed by the need to extract oxygen from water, an O₂-poormedium; thus, acid-base balance is maintained primarily throughion transport mechanisms. These mechanisms include Na⁺/H⁺ andCl^–/HCO₃^– exchange processes that are sensitiveto the extracellular acid-base status of the animal. In marinecrabs, ion regulation and acid-base balance are accomplishedby the posterior gills, while in freshwater species all gillsand the antennal gland perform these functions. Intracellularacid-base balance appears to be maintained primarily by iontransport across the cell membrane. Hemolymph pH varies inverselywith acclimation temperature and salinity. In both cases Pco₂remains nearly constant, and the pH change is a result of changesin hemolymph HCO₃^– concentrations brought about by ionexchange mechanisms. Environmental hypercapnia or hyperoxiainduces a repiratory acidosis characterized by increased Pco₂,low pH, and elevated HCO₃^–; this is partially compensatedfor by ion exchange processes that bring about a further increasein hemolymph HCO₃^–. Exercise causes a mixed respiratoryand metabolic acidosis with compensation via H⁺ ion excretionand hyperventilation.     

CO2 Excretion Across Isolated Perfused Crab Gills: Facilitation by Carbonic Anhydrase

Crab gill carbonic anhydrase is shown to facilitate the excretionof carbondioxide across isolated perfused gills. A techniquefor perfusing crab gills and assessing the metabolic viabilityof perfused gills is also described in detail. The techniqueis used to follow the disappearance of ¹⁴C label as HCO₃^–and CO₂ from internal perfusate passing through the gill. Theexcretion of the label increases with the flow rate of the externalperfusate across the outside of the gills. The addition of carbonican hydrase to the internal perfusate results in a two- to fourfoldincrease in the excretion of label while Diamox (acetazolamide)treatment decreases the excretion of label by half. It is alsosuggested that carbonic anhydrase, present in muscle tissuesof crabs, minimizes the disequilibrium of the hemolymph CO₂system as metabolically produced CO₂ leaves the tissues andenters the hemolymph. Parallels are drawn between the presenceof carbonic anhydrase in the crab gill system and the presenceof this enzyme in the respiratory organs of both aquatic andterrestrial animals.     

Effect of Oxygen on Photosynthetic 14CO2 Fixation in Chroomonas sp. (Cryptophyta) I. Some Characteristics of the Oxygen Effect

The effect of oxygen on photosynthetic ¹⁴CO₂ fixation in theair-grown freshwater flagellate Chroomonas sp. (Cryptophyta)was studied. Considerable inhibition by anaerobiosis was observedonly under light-saturated conditions with no effect from theCO₂ concentration. This inhibition was reversed by 2% O₂. With,more than 2% O₂, the rate of ¹⁴CO₂ fixation was inhibited; 100%O₂ caused about 20% inhibition which could be reversed by 2%O₂. The degree of inhibition was only slightly higher at lowconcentrations (less than 0.43 mM NaHCO₃) than at high CO₂ concentrations,indicating that photorespiration is not a main cause of thisinhibition. Possible causes of the inhibitions by anaerobiosisand by oxygen are discussed. (Received June 29, 1983; Accepted January 23, 1984)     

Acid-Base Regulation and Temperature in Selected Invertebrates as a Function of Temperature

The pH of the hemolymph of selected invertebrates decreasesas their body temperature increases. The magnitude of this change(pH/°C) is very similar to the change of the pH of waterwith temperature (pN/°C) and suggests that these invertebrates,like poikilothermous vertebrates, regulate the pH of their extracellularfluid so that its degree of alkalinity relative to the pH ofwater remains constant. The degree of alkalinity (pH_blood-pN)varies between species, but seems to be fixed for any givenspecies. In Limulus pH-pN was essentially the same for in vivosamples, measured after the whole animal had been acclimatedto different temperatures, as it was for in vitro samples inwhich the hemolymph was cooled or warmed anaerobically, suggestingthat the CO₂ content of the extracellular fluid is constantas the temperature changes. The P_CO2 of the hemolymph is invariablylower in animals breathing water than in those breathing air.In the invertebrates, as in the vertebrates, manipulation ofP_CO2 and HCO₃- is probably the major mechanism in the regulationof the relative alkalinity of the extracellular fluid.     

Physiological responses to emersion in the intertidal green crab,Carcinus maenas (L.)

The green shore crab, Carcinus maenas, undergoes on average 6?h periods of emersion during each low-tide cycle during the summer months. Under those conditions, the crab is cut off from its normal water environment and is exposed to potential stress from a suite of environmental and physiological changes: dehydration, compromised gas exchange and resultant internal hypoxia and hypercapnia, thermal stress, and ammonia toxicity. This study examined the comprehensive responses of the green crab in water and to a 6?h emersion period laboratory simulation of a tidal cycle followed by a 1?h re-immersion period, measuring indicators of dehydration, hemolymph osmolality, oxygen uptake, hemolymph acid–base status, heart and ventilatory rate, and hemolymph ammonia and ammonia excretion. Green crabs showed physiological responses of varying magnitude to 6?h of emersion. Individuals were found in the field exclusively under rocks and large clumps of seaweed where temperatures were approximately half those of exposed surfaces and relative humidity was about twice as high as ambient air. During emersion, crabs lost less than 5% of their wet weight, and hemolymph osmolality did not increase significantly. Oxygen uptake continued in air at about 50% of the control, aquatic values; and the gills continued to be ventilated by the scaphognathite, albeit at lower rates. Hemolymph lactate concentrations increased, indicating a shift to a greater reliance on anaerobic metabolism to support energetic needs. A slight acidosis developed in the hemolymph after 1?h of emersion, but it did not increase thereafter. Ammonia concentrations in the hemolymph were unchanged, as ammonia was volatilized by the gills and excreted into the air as NH₃ gas. These results show that the green crab copes with emersion by seeking refuge in microhabitats that mitigate the changes in the physical parameters of intertidal emersion. Physiologically, desiccation is avoided, cardio-respiratory processes are maintained at reduced levels, and hemolymph acid–base balance is minimally affected. Ammonia toxicity appears to be avoided by a shift to excreting NH₃ gas directly or indirectly to air.     

Estimation of Photorespiration of Green Plants and of their Mesophyll Resistance to CO2 Uptake

An estimate of photorespiration is obtained from the relationshipbetween the net exchange of CO₂ of the leaf and the internalCO₂ concentration, i.e. within the mesophyll intercellular spaces.The latter is obtained by calculation, taking into account thecombined epidermal and boundary-layer resistances between thebulk atmosphere and the mesophyll intercellular spaces. Thelinear part of this relationship (at low CO₂ concentrations)is extrapolated to zero internal concentration, at which noneof the intercellular photorespired CO₂ is available for reassimilation.The calculated output of CO₂ under such conditions providesan estimate of photorespiration, but, by failing to take intoaccount intracellular reassimilation of photorespired CO₂ underestimatesactual photorespiration. As the slope of this linear relationshiprepresents the mesophyll (intracellular) resistance to CO₂ uptake,this procedure was used to recalculate published data on effectsof light intensity and of oxygen concentration on net photosynthesis.The analysis showed that increased oxygen concentration anddecreased light intensity reduced photosynthesis largely byincreasing mesophyll resistance to CO₂ uptake. It is suggestedthat the CO₂ compensation point () is a function of both photorespiration(L) and mesophyll resistance (r_m): = L. r_m.     

Dormancy in Cereal Seeds: II. THE NATURE OF THE GASEOUS EXCHANGE IN IMBIBED BARLEY AND RICE SEEDS

When barley seeds imbibe water, the O₂ uptake of non-dormantseeds is considerably less than that of dormant seeds for atleast the first 6 h, irrespective of the rate at which the seedshad previously lost dormancy. During the initial 6 h of imbibition, the CO₂ output of dormantbarley seeds is usually only slightly greater than and sometimesno different from that of nondormant seeds. The CO₂ output ofdormant seeds is reduced by about 66 percent by millimolar KCN,whereas that of non-dormant seeds is decreased by about 12–13per cent only. The CO₂ output of dormant barley in nitrogenis considerably less than the CO₂ output of non-dormant seedsunder the same conditions. Dormant rice seeds also show a higher initial O₂ uptake thannon-dormant seeds, though this is not generally as marked asin barley. Similarly, the initial CO₂ output of dormant seedsis distinctly greater than that of non-dormant seeds, but inmillimolar KCN it is depressed to a greater extent than in non-dormantseeds. In nitrogen, the CO₂ outputs of dormant and non-dormantseeds were found to be the same. Consequently, unlike barley,dormant rice seeds appear to be as capable of carrying out alcoholicfermentation under anaerobic conditions as nondormant seeds. In barley, increasing the O₂ tension from 21 per cent to 100per cent increased the oxygen uptake of dormant seeds more thanthat of non-dormant seeds (an increase of 53 per cent as against20–23 Per cent). In dormant seeds there was a concomitantincrease in CO₂ output (about 50 per cent), but the CO₂ outputof non-dormant seeds was hardly affected. High concentrations of CO₂ are inhibitory to the germinationof both dormant and non-dormant barley seeds. At a concentrationof 10 per cent, however, CO₂ is inhibitory only to dormant seeds,although at 2.5–5 per cent it is sometimes stimulatoryto the germination of dormant seeds. A 24–h treatmentwith appropriate concentrations of ethanol, lactic acid, oracetaldehyde is also stimulatory to the germination of dormantbarley seeds. Histochemical investigations in barley indicated the presenceof peroxidase, cytochrome oxidase, and -glycero-phosphate dehydrogenasein the embryo, aleurone layer, and in a layer associated withthe testa. A number of other redox enzymes were detected inthe embryo and aleurone layer only. No differences in distributionor intensity of activity were detected between dormant and nondormantseeds.     

Effect of carbon dioxide concentration on pigmentation in the blue-green alga Anacystis nidulans

The pigment content in the blue-green alga Anacystis nidulanswas found to be dependent upon CO₂ concentration during growth.In cells grown with 1% CO₂ in air the total pigment constituted20.5% of the dry weight while it was only 11.1% of dry weightof cells grown in air (0.03% CO₂). This decrease in total pigmentwas found to be almost entirely ascribable to decrease in phycocyanin.Since light absorbed by phycocyanin has been shown to providenearly equal rates of photoreactions I and II, the "CO₂ control"of phycocyanin is viewed as an effective means of regulationof the photoreactions without upsetting the balance of operationof the two photoreactions. (Received December 25, 1970; )     

Elevated carbon dioxide has the potential to impact alarm cue responses in some freshwater fishes

Freshwater fish behaviors have the potential to be impacted by acidification due to increases in dissolved carbon dioxide (CO₂). Recent work in the marine environment suggests that increased CO₂ levels due to climate change can negatively affect fishes homing to natal environments, while also hindering their ability to detect predators and perform aerobically. The potential for elevated CO₂ to have similar negative impacts on freshwater communities remains understudied. The objective of our study was to quantify the effects of elevated CO₂ on the behaviors of fathead minnows (Pimephales promelas) and silver carp (Hypophthalmichthys molitrix) following exposure to conspecific skin extracts (alarm cues). In fathead minnows, their response to conspecific skin extracts was significantly impaired following exposure to elevated CO₂ levels for at least 96 h, while silver carp behaviors were unaltered. However, fathead minnow behaviors did return to pre-CO₂ exposure in high-CO₂-exposed fish following 14 days of holding at ambient CO₂ levels. Overall, this study suggests there may be potential impacts to freshwater fishes alarm cue behaviors following CO₂ exposure, but these responses may be species-specific and will likely be abated should the CO₂ stressor be removed.     

Physiological response to emersion in the amphibious crab Chasmagnathus granulata Dana (Decapoda Grapsidae): biochemical and ventilatory adaptations

The adaptation of Chasmagnathus granulata to air breathing depends on two types of physiological mechanisms: (1) Biochemical adjustments, comprising the achievement of new steady state values for partial pressure of carbon dioxide (P_CO2) and total carbon dioxide concentration (C_CO2). The initial increase in hemolymph bicarbonate is stabilized by dehydration to CO₂ presumably catalyzed by carbonic anhydrase (CA) at the basolateral side of the gill epithelium. Thus, an adequate transbranchial gradient of P_CO2 is restored. Inhibition of CA with acetazolamide (Az) reduces the rate of CO₂ excretion and elevates P_CO2. The respiratory acidosis caused by increased P_CO2 is compensated by increase in the difference between Na⁺ and Cl⁻ concentrations. Az does not affect hemolymph ionic concentrations. (2) Ventilatory control: untreated animals show a significant decrease in scaphognathite frequency (F_SC) during emersion, while Az treated crabs show a slight increase of this variable. F_SC of Az crabs tends to decrease with hemolymphatic CO₂, presumably by clearance of the CA inhibitor from hemolymph. These results suggest that C. granulata possess a ventilatory control based on a primary oxygen-dependent stimulus and a secondary one dependent on CO₂.     

Biosynthetic mechanism of glycolate in Chromatium 6. Glycolate formation and metabolism under low O21

Under low O₂ (0.05 mM O₂), there was no measurable excretionof glycolate or glycine by Chromatium cells, unlike the caseof their incubation under high (0.7 mM) O₂ However, upon additionof non-radioactive glycolate and glycine to the suspension medium,there occurred a measurable incorporation of ¹⁴CO₂ into thesecompounds, which were then excreted extracellularly; the totalradioactivities measured were approximately 15% of the totalCO₂ fixed photosynthetically. This phenomenon could be as cribedto the dilution of the intracellular pools by the compoundsadded. The results indicate that under low O₂ the glycolatemolecules produced are metabolically further transformed inthe bacterial cells. The incorporation of ¹⁴CO₂ into the extracellularglycolate fraction was maximal at 0.3 mM glycolate in both highand low O₂. Presumably, glycolate formed in the bacterial cellsunder both the high and low O₂ is metabolized in a similar manner,although the excess glycolate and glycine molecules are rapidlyexcreted. During glycolate metabolism CO₂ was evolved from anisonicotinylhydrazide-sensitive reaction, suggesting that thepathway <glycolate glycine . CO₂ was similar in green plants.The results thus indicate that studies on glycolate and glycinemetabolism in the anaerobic bacterium, Chromatium, provide auseful model system for elucidating the mechanism of photorespirationin green plants. (Received May 19, 1978; )     

Effects of Carbon Dioxide on Metabolism by the Glycollate Pathway in Leaves

When solutions of [¹⁴C]glycollate, glycine, serine, glycerate,or glucose were supplied to segments of wheat leaves throughtheir cut bases in the light, most of the ¹⁴C was incorporatedinto sucrose in air but in CO₂-free air less sucrose was made.The synthesis of sucrose was decreased because metabolism ofserine was partly blocked. Sucrose synthesis from glucose andglycerate in CO₂-free air was decreased but to a smaller extent;relatively more CO₂ was evolved and serine accumulated. Theeffects of DCMU and light of different wavelengths on metabolismby leaves of L-[U-¹⁴C]serine confirmed that simultaneous photosyntheticassimilation of carbon was necessary for the conversion of serineto sucrose. Of various products of photosynthesis fed exogenouslyto the leaves -keto acids were the most effective in promotingphotosynthesis of sucrose and release of ¹⁴CO₂ from ¹⁴C-labelledserine. This suggests that in CO₂-free air the metabolism ofserine may be limited by a shortage of -keto acid acceptorsfor the amino group. In CO₂-free air added glucose stimulatedproduction of CO₂ and sucrose from D-[U-¹⁴C]- glycerate andno competitive effects were evident even though glucose is convertedrapidly to sucrose under these conditions. In addition to asupply of keto acid, photosynthesis may also provide substratesthat can be degraded and provide energy in the cytoplasm forthe conversion of glycerate to sugar and phosphates and sucrose.     

Respiration and Circulation in Land Crabs: Novel Variations on the Marine Design

Both the "true" crabs (Brachyura) and hermit crabs (Anomura)include species that show numerous behavioral, morphological,and physiological specializations permitting terrestrial life.This paper examines respiratory and circulatory adaptationsfor air breathing in these land crabs. Respiratory specializationsinclude modification of gas exchange structures for air breathing(gills and elaborated branchial chamber linings), ventilatorymechanisms permitting effective air pumping, an elevated hemolymphoxygen capacity, and a primarily CO₂- rather than O₂- sensitiveventilatory control system. The qualitative aspects of hemolymphoxygen transport and metabolic rate are apparently unchangedfrom that of marine crabs. While the basic cardiovascular morphologyof land crabs appears similar to that of marine forms, thereis considerable elaboration of the vasculature of the branchialchamber lining, which in some species includes a unique doubleportal system. Cardiac output is lower in land crabs (probablyrelated to their higher hemolymph O₂ capacity), but insufficientdata on hemolymph pressures prevent comparisons with marineforms. In general, land crabs have modified (sometimes extensively)existing structures and processes found in their marine relativesrather than evolving structures for terrestrial life de novo.Accordingly, land crabs present a useful model for the evolutionof terrestriality, showing that even subtle anatomical changescan result in the large changes in physiological function necessaryfor the terrestrial invasion.     

The Metabolism of Acetaldehyde by Plant Tissues

Apples and oranges can absorb appreciable amounts of acetaldehydevapour, but acetaldehyde does not accumulate in the tissues,in the presence or absence of oxygen. When the fruit is givenacetaldehyde the loss of carbon as CO₂ + ethanol equals thatlost from carbohydrate + acid+acetaldehyde. Part of the addedacetaldehyde is reduced to ethanol by NADH₂ and the rate ofglycolysis is increased; the remainder is oxidized to CO₂. Respiration of apples is limited by oxygen; the output of CO₂is only slightly increased at the beginning of the treatmentwith acetaldehyde, but afterwards it declines; acetaldehydespares oxidation of carbohydrate. In oranges, availability of substrate limits respiration, andacetaldehyde stimulates CO₂-production and O₂-uptake. In theabsence of oxygen, acetaldehyde is reduced to ethanol, and thequotient CO₂/ethanol falls.     

Oxygen enhancement of photosynthesis in Anacystis nidulans cells1

Oxygen enhanced photosynthetic ¹⁴CO₂ fixation in Anacystis nidulanscells. Results obtained under different conditions revealedthe following properties of the oxygen enhancement:

1. The enhancement was most significant at ca. 10% O₂. Furtherincrease in oxygen concentration decreased the enhancing effect.The rate under 100% O₂ was equivalent to or a little higherthan that under N₂ gas.
2. b) With the increase in CO₂ concentration,the magnitude ofthe enhancing effect decreased. No oxygen enhancementwas observedwhen the CO₂ concentration. was raised to 9,000ppm.
3. c) The enhancement was observed only at high light intensities.No enhancement was observed when the rate of photosynthesiswas limited by light intensity.
4. Ribulose 1,5-diphosphate (RuDP)carboxylase activity was demonstratedin the extract obtainedfrom A. nidulans cells. We also foundthat the RuDP carboxylaseactivity in this extract was competitivelyinhibited by oxygen.
5. Based on the above-mentioned results, the possible mechanismunderlying the observed enhancing effect of oxygen was discussed.

(Received May 10, 1976; )     

Activation of Glyoxylate Cycle Enzymes in Cucumber Fruits Exposed to CO2

Cucumber fruits exposed to a 60% CO₂ environment for 3 d at20°C showed a higher respiration rate, a lower respiratoryquotient, and lower levels of malate, citrate + isocitrate,and glyoxylate than did control fruits. Moreover, in the CO₂treatment, fumarate and succinate increased dramatically onthe first day and after the second day, respectively, while2-oxoglutarate showed a temporary increase on the first dayand a subsequent decrease. Isocitrate lyase and malate synthaseactivity tended to increase throughout the experimental periodof the CO₂ treatment. These results appear to point to a rapidoperation of glyoxylate cycle due to high CO₂. (Received November 14, 1997; Accepted March 2, 1998)     

Oxygen Exchange in Relation to Carbon Assimilation in Water-stressed Leaves During Photosynthesis

In a study on metabolic consumption of photosynthetic electronsand dissipation of excess light energy under water stress, O₂and CO₂ gas exchange was measured by mass spectrometry in tomatoplants using ¹⁸O₂ and ¹³CO₂. Under water stress, gross O₂ evolution(E_O), gross O₂ uptake (U_O), net CO₂ uptake (P_N), gross CO₂ uptake(TPS), and gross CO₂ evolution (E_C) declined. The ratio P_N/E_Ofell during stress, while the ratios U_O/E_O and E_C/TPS rose.Mitochondrial respiration in the light, which can be measureddirectly by ¹²CO₂ evolution during ¹³CO₂ uptake at 3000 µll^{–1 13}CO₂, is small in relation to gross CO₂ evolutionand CO₂ release from the glycolate pathway. It is concludedthat PSII, the Calvin cycle and mitochondrial respiration aredown-regulated under water stress. The percentages of photosyntheticelectrons dissipated by CO₂ assimilation, photorespiration andthe Mehler reaction were calculated: in control leaves morethan 50 % of the electrons were consumed in CO₂ assimilation,23 % in photorespiration and 13 % in the Mehler reaction. Undersevere stress the percentages of electrons dissipated by CO₂assimilation and the Mehler reaction declined while the percentageof electrons used in photorespiration doubled. The consumptionof electrons in photorespiration may reduce the likelihood ofdamage during water deficit.     

Differences between estimates and measured PaCO2 during rest and exercise in older subjects

Williams, J. S., and T. G. Babb. Differences betweenestimates and measured Pa_CO2 during restand exercise in older subjects. J. Appl.Physiol. 83(1): 312-316, 1997.ArterialPCO₂ (Pa_CO2) has been estimated duringexercise with good accuracy in younger individuals by using the Jonesequation(P_JCO₂)(J. Appl. Physiol. 47: 954-960,1979). The purpose of this project was to determine the utility ofestimating Pa_CO2 from end-tidal PCO₂(PET_CO2) orP_JCO₂at rest, ventilatory threshold (Th), and maximalexercise (Max) in older subjects. PET_CO2 was determined fromrespired gases simultaneously (MGA 1100) with arterial blood gases(radial arterial catheter) in 12 older and 11 younger subjects at restand during exercise. Mean differences were analyzed with pairedt-tests, and relationships between theestimated Pa_CO2 values and the actualvalues of Pa_CO2 were determined withcorrelation coefficients. In the older subjects, PET_CO2 was not significantlydifferent from Pa_CO2 at rest (1.2 ± 4.3 Torr), Th (0.4 ± 2.5), or Max(0.8 ± 2.7), and the two were significantly(P < 0.05) correlated atth (r = 0.84) andMax (r = 0.87) but not atrest (r = 0.47).P_JCO₂was similar to Pa_CO2 at rest (1.0 ± 3.9) and th (1.3 ± 2.3) but significantly lower at Max (3.0 ± 2.6), and the two weresignificantly correlated at th(r = 0.86) and Max(r = 0.80) but not at rest (r = 0.54).PET_CO2 was significantlyhigher than Pa_CO2 during exercise in theyounger subjects but similar to Pa_CO2 at rest.P_JCO₂was similar to Pa_CO2 at rest andth but significantly lower at Max in youngersubjects. In conclusion, our data demonstrate thatPa_CO2 during exercise is betterestimated by PET_CO2 than byP_JCO₂in older subjects, contrary to what is observed in younger subjects.This appears to be related to the finding thatPET_CO2 does not exceedPa_CO2 during exercise in older subjects,as occurs in the younger subjects. However,Pa_CO2 at rest is best estimated byP_JCO₂in both younger and older subjects.

     

Phytoplanktonic carbon isotope fractionation: equations accounting for CO2-concentrating mechanisms

A model of carbon isotope discrimination by phytoplankton wasdeveloped which took into account the occurrence of a carbon-concentratingmechanism (CCM). A simple equation was obtained for the modelinvolving CO₂ active transport. In the case of HCO₃^– activetransport, another equation was developed based on a seriesof approximations. The former equation was used to analyse reportedand newly obtained data from culture experiments and field observationsin both freshwater and marine environments. In most cases, alinear relationship between a combined parameter, (1–f)Ci,which was made up of the relative contribution of active CO₂uptake to total carbon uptake (f) and the intracellular CO₂concentration (Ci), and CO₂ concentration in bulk solution (Ce)was obtained as (1–f)Ci = ace–b, with a high correlationcmfficient (r²>0.9). The slope a is suggested as a measureof the ratio of diffusive to total (diffusive+active) CO₂ transport,while bla represents CO₂ demand.