Changes in the blood parameters of an air-breathing fish during different respiratory conditions.

Some of the blood parameters recorded in an air-breathing eel, Amphipnous cuchia under normal respiratory condition during non-breeding period (September-April) are haemoglobin (Hb) concentration 19.26%, haematocrit value 56.16%, RBC number 1.71 million/mm3, RBC size 18.86 X 9.70 mum, mean corpuscular haemoglobin (MCH) 113.4 ng, mean corpuscular haemoglobin concentration (MCHC) 34.2%, blood sugar 77 mg% and ascorbic acid 0.435 mg%. The higher concentration of haemoglobin (19.26%) appears to be related to its obligatory air breathing habit and habitat in a water of low oxygen content. Though a definite trend of increase in the haemoglobin and haematocrit concentration with an increase in the body weight of the fish was lacking, variations were clearly marked related to intrinsic activity of the fish connected with different respiratory conditions. Asphyxiation in a submerged but continuous flow of water (liter/h) for 5 1/2 h resulted in an increase in the above-mentioned parameters to an appreciable extent. These increases were 0.23 million/mm3 in the number of erythrocytes, 6.16% in haemoglobin concentration, 10% in haematocrit value, 20% in blood sugar and 35% in ascorbic acid content. The mean corpuscular haemoglobin showed a decline of 6.2%. Exclusive aerial breathing for 5 1/2 h also caused 7.4% increase in haemoglobin concentration, 9.4% in haematocrit value, 0.14 million/mm3 in RBC number, 20% in blood sugar level, 9% in ascorbic acid content but almost no change in mean corpuscular haemoglobin. The average surface area for diffusion of gases appeared to have reduced by 6.8 mum2 per RBC.     

Blood adaptations to marine and freshwater environments in fish of the Family Sciaenidae (Perciformes)

In five species of fish from the Family Sciaenidae, collected from marine, brackish and fresh-water environments, the following parameters were studied: haemoglobin concentration, haematocrit, number of red blood cells, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, blood pH, oxygen affinity of blood and stripped haemolysate, Root effect, the number of haemoglobins separated by polyacrilamide electrophoresis, iron concentration and osmotic pressure of the serum. Plagioscion squamosissimus , the only freshwater species studied, clearly separated from the other four species; it exhibited the highest haemoglobin number, haemotocrit, number erythrocytes, of oxygen affinity of the haemolysate, and the lowest oxygen affinity of the blood, iron concentration, pH and osmotic pressure.     

An Evolutionary Perspective for Bimodal Respiration: A Biological Synthesis of Fish Air Breathing

SYNOPSIS: The broad phyletic distribution of bimodal breathingamong the fishes, the diversity of this group's aerial-respiratoryspecializations, and the numerous ways that bimodality has permeatedthe natural history of many species provide a broad perspectiveon the evolution and biological significance of bimodal breathing.The long term evolutionary view of bimodal breathing emphasizesits importance in the evolutionary transition to terrestriality;a key role in this process was played by the fishes which werethe first air-breathing vertebrates. On the other hand, in mostextant fishes bimodality has not led to terrestriality. Rather,auxiliary air breathing enables a species to remain in or toexploit an aquatic habitat from which it would otherwise beexcluded.     

Red blood cell function and haematology in two tropical freshwater fishes from Australia

Salmon catfish and tarpon occur in habitats that periodically become deficient in oxygen resulting in high mortalities of other fish species. The water-breathing catfish, Arius leptaspis, and the facultative air-breathing tarpon, Megalops cyprinoides, both have high haemoglobin and haematocrit, and the oxygen carrying capacity in the air-breather is exceptionally high (15.6+/-1.2 vol%). Iso-pH oxygen equilibria of the red blood cells at 25 degrees C revealed high affinity (P(50)=9 mmHg, pH 7.4) and co-operativity (n(50)>2.2, pH 7.4) in the catfish, and contrasted with low affinity (P(50)=32 mmHg, pH 7.4) and co-operativity (n(50) approximately 1) in the air-breathing tarpon. Oxygen binding was further distinguished by relative pH insensitivity (Bohr factor, ?=Deltalog P(50)/Deltalog pH=-0.22) in the catfish, compared with a significant Bohr effect in the tarpon (?=-0.96). The potential for modulation of haemoglobin-oxygen affinity was indicated by a high ratio of GTP to ATP in the erythrocytes of the catfish, whereas regulation in the tarpon appeared due to ATP alone. Differences in blood respiratory functions between the two species are likely to reflect reduced opportunity for activity under extreme hypoxia in the catfish.     

A functional analysis of the shift from gill- to lung-breathing during the evolution of land crabs (Crustacea, Decapoda)

The evolution of air-breathing in land crabs is associated with a progressive shift in the primary site of respiratory gas exchange from the diffusion-limited gills used for water-breathing, via a simple 'cutaneous' lung surface to the perfusion-limited, invaginated lung described in the mountain crab, Pseudothelphusa garmani. The reduced diffusion limitation over the lungs facilitates oxygen transfer from air to the tissues at lower ventilation rates but is associated with accumulation of carbon dioxide. A potential respiratory acidosis is buffered by the respiratory pigment haemocyanin and by elevation of haemolymph bicarbonate levels. These changes parallel those described in vertebrates but air-breathing crustaceans maintain relatively low carbon dioxide levels in the haemolymph, either by retaining an aquatic route for its elimination over the reduced gills or by blowing it off across the lung. Maintenance of low carbon dioxide levels may be associated with a limited capacity to buffer against an acidosis due to low levels of circulating haemocyanin (i.e. crustaceans lack red blood cells). This may ultimately limit their survival in air as an acidosis will reduce oxygen transport due to a marked Bohr effect on haemocyanin. The primary role of an invaginated lung may be to reduce rates of water loss in air.     

The Respiratory Transition from Water to Air Breathing During Amphibian Metamorphosis

SYNOPSIS. Profound developmental changes occur in the morphologyand physiology of the respiratory system of amphibians duringthe transition from strictly aquatic to dual aquatic-aerialbreathing. This developmental transition usually involves modificationsin ventilatory mechanisms and/or respiratory surfaces {e.g.,degeneration of gills, ventilation of functional lungs). Boththe first appearance of obligate air breathing and the subsequentdependence upon it by amphibian larvae are sensitive to a varietyof environmental stressors during critical developmental periods.These stressors include oxygen availability, ambient temperature,the risk of predation and mode of feeding.     

Respiratory Control in the Transition from Water to Air Breathing in Vertebrates

SYNOPSIS. Studies on extant bimodally breathing vertebratesoffer us a chance to gain insight into the changes in respiratorycontrol during the evolutionary transition from water to airbreathing. In primitive Actinopterygian air-breathingfishes(Lepisosteus and Amid), gill ventilation is driven by an endogenouslyactive central rhythm generator that is powerfully modulatedby afferent input from internally and externally oriented branchialchemoreceptors, as it is in water-breathing Actinopterygians.The effects of internal or external chemoreceptor stimulationon water and air breathing vary substantially in these aquaticair breathers, suggesting that their roles are evolutionarilymalleable. Air breathing in these bimodal breathers usuallyoccurs as single breaths taken at irregular intervals and isan on-demand phenomenon activated primarily by afferent inputfrom the branchial chemoreceptors. There is no evidence forcentral CO₂/pH sensitive chemoreceptors and air-breathing organmechanoreceptors have little influence over branchial- or air-breathingpatterns in Actinopterygian air breathers. In the Sarcopterygianlungfish Lepidosiren and Protopterus, ventilation of the highlyreduced gills is relatively unresponsive to chemoreceptor ormechanoreceptor input. The branchial chemoreceptors of the anteriorarches appear to monitor arterialized blood, while chemoreceptorsin the posterior arches may monitor venous blood. Lungfish respondvigorously to hypercapnia, but it is not known whether theseresponses are mediated by central or peripheral chemoreceptors.A major difference between the Sarcopterygian and Actinopterygianbimodal breathers is that lungfish can inflate their lungs usingrhythmic bouts of air breathing, and lung mechanoreceptors influencethe onset and termination of these lung inflation cycles. Thecontrol of breathing in amphibians appears similar to that oflungfish. Branchial ventilation may persist as rhythmic buccaloscillations in most adults, and stimulation of peripheral chemoreceptorsin the aortic arch or carotid labyrinths initiates short boutsof breathing. Ventilation is much more responsive to hypercapniain adult amphibians than in Actinopterygian fishes because ofcentral CO₂/pH sensitive chemoreceptors that act to convertperiodic to more continuous breathing patterns when stimulated.     

Morphological, Behavioral, and Physiological Characterization of Bimodal Breathing Crustaceans

SYNOPSIS Bimodal breathing crustaceans, while representing astage in the transition from the aquatic to terrestrial habitat,also constitute a distinct group that can be characterized bymorphological, behavioral, and physiological traits. Morphologically,this group displays reduced gill surface area and enlarged branchialchambers. The lining of the branchial chambers, the branchiostegites,also has increased surface area and is highly vascularized.The branchiostegites can be smooth, cutaneous epithelia, orthey can have more complex evaginations or invaginations tofurther increase surface area. In addition, the thickness ofthe branchiostegal epithelium is greatly reduced, compared tothat in the gills, thus minimizing the diffusion distance betweenair and hemolymph. These animals maintain a store of water inthe branchial chamber that covers the gills and allows for simultaneousgas exchange with two media (air and water). There is also apartitioning of gas exchange, with oxygen uptake occurring preferentiallyfrom air across the branchiostegites, and carbon dioxide excretionoccurring across the gills into the branchial water. One criticalfactor that appears to separate bimodal breathing crustaceansfrom fully terrestrial, exclusively air-breathers is the abilityof the latter group to excrete CO₂directly into air across thegills and branchiostegites. It is suggested that the incorporationof the enzyme carbonic anhydrase into the membrane fractionof the branchiostegites may have been one of the key molecularevents which allowed for pulmonary CO₂ excretion into air.     

Basic haematological values in antelopes--IV. A comparison and concluding remarks

Comparison of mean erythrocyte counts, mean haematocrit values, mean haemoglobin content and mean corpuscular volume of the red cell is presented for 21 species of antelopes (four species being excluded because of the small number of animals investigated). A direct relationship of haematocrit values and haemoglobin content and an inverse relationship of red cell counts and mean corpuscular volume of the erythrocytes were noted. The significance of an increased total surface area for oxygen exchange is discussed and values obtained in the red blood picture are compared with those of domesticated animals (cattle, goat and sheep) taken from the literature. Mean leukocyte counts were found to be in the human range with the exception of two species, but significantly lower than in domesticated artiodactylids. Problems in assessing the general health, the age, the effect of diet and of environment in captivity are discussed. Problems of methodology, especially of blood sampling, are given comparing results in manually restrained and in sedated animals.     

Observations on the haematology and oxygen transport of the New Zealand fur seal,Arctocephalus forsteri

Abstract

Red cell count, haemoglobin concentration, haematocrit, mean cell volume, and mean cell haemoglobin concentration were recorded for the fur seal Arctocephalus forsteri (Lesson). The data did not indicate haematological adaptations for deep diving nor for extended periods of submergence. Two distinct haemoglobin types were isolated from the red cells by electrophoresis. The oxygen affinity of the blood was low as measured by half-saturation values (p50) of 42.3 mm Hg at pH 7.1 and 26.2 mm Hg at pH 7.4 and 37°c. The low oxygen affinity was mediated by erythrocytic 2,3-diphosphoglycerate, and on this basis a high turnover of oxygen to the tissues is postulated. The role of the blood in oxygen transport appears to be suited for feeding near the surface rather than by deep diving.     

Respiratory,cardiovascular, and hemolymph acid‐base changes in the amphibious crab,Cardisoma guanhumi,during immersion and emersion

The cardiorespiratory and hemolymph acid base status of bimodal breathing crabs, Cardisoma guanhumi, was monitored during the transition from breathing air to breathing water. Upon immersion, oxygen uptake (MO₂) decreased by half. Ventilatory frequency (fsc) increased more than 5 fold, causing a decrease in hemolymph carbon dioxide partial pressure (PCO₂). This was nearly fully compensated for by a gradual decrease in hemolymph bicarbonate concentration ([HCO₃ ^‐]) over 96 hours post‐immersion. After one to two weeks of immersion, when crabs were removed from the water, oxygen uptake initially increased, but eventually returned to the initial immersed value. Heart rate was unchanged but fsc slowed dramatically. The decreased ventilation resulted in a buildup of hemolymph PCO₂, causing a respiratory acidosis that was slowly compensated for by increased hemolymph [HCO₃ ^‐]. C. guanhumi appears to be a truly amphibious crab with respiratory and acid‐base adaptations found in both fully aquatic and fully terrestrial species.     

Respiratory functions of the blood in the limpet Siphonaria zelandica (Gastropoda: Pulmonata)

Abstract

The respiratory pigments myoglobin and haemocyanin were characterised in the marine pulmonate Siphonaria zelandica (Quoy & Gaimard) and their roles in an oxygen transfer system were postulated. In air, when the animals were active, oxygen was transported from a simple diffusion lung by haemocyanin in the blood which had a half-saturation value of 12.7 mm Hg at pH 7.2 and 25°c. At pH 7.6 the oxygen affinity decreased to 17.3 mm Hg, indicating a reverse Bohr effect which might be expected to facilitate oxygen uptake in the lung during bursts of activity at low tide. A high oxygen-combining capacity of buccal mass myoglobin (21.2 vols %) indicated a role of oxygen storage during bursts of feeding activity. The distribution of carbonic anhydrase in various tissues was consistent with a transfer system facilitating the release of metabolic carbon dioxide from the buccal mass.     

Oxygenation properties of haemoglobins from the flatfish plaice (Pleuronectes platessa) and flounder (Platichthys flesus)

Summary The oxygen binding properties and some haematological data of haemoglobins of plaice (Pleuronectes platessa) and flounder (Platichthys flesus) were measured and compared, with the view of discerning their adaptations to the benthic habitat.Compared to plaice, flounders posses higher haematocrit and haemoglobin levels, and their haemoglobin has a higher oxygen affinity, smaller Bohr and Root effects and appears to have a greater ATP sensitivity. At pH 7.6, the half-saturation oxygen tensions,P ₅₀, of stripped plaice and flounder haemoglobins amount to about 5.9 and 4.2 torr, and the Bohr shifts (logP ₅₀/ pH) to –0.51 and –0.34, respectively. The species differences in the oxygen binding patterns occur after stripping the haemoglobin in solution, from dissolved ions and thus appear to be inherent in the pigment molecules. The differences suggest that the haemoglobin of flounder is better adapted to hypoxic and hypercarbic environments than plaice haemoglobin, in accordance with the more inshore occurrence of the former species.The molar ratios of ATP to haemoglobin tetramers are low in plaice and flounder (about 1.6); in plaice this ratio corresponds to that where the cofactor effect is most pronounced. In contrast to previous evidence for an adaptive reduction in the temperature dependence of the haemoglobin of flounder but not of plaice from the Baltic Sea (Friedrich, 1935), the same temperature effects were found in representatives of the two species both from the Dutch North Sea and the Danish Kattegat.     

Episodic Breathing in Frogs: Converging Hypotheses on Neural Control of Respiration in Air Breathing Vertebrates

SYNOPSIS. The episodic, or intermittent, breathing of frogsand many ectothermic vertebrates results in important fluctuationsof arterial blood gases. This pattern of breathing differs fromthe rhythmic and continuous alternation of inspiration observedin most homeotherms, which maintain O₂ and CO₂ levels withinnarrow ranges. These differences in pattern of breathing indicatethat the respiratory control systems of ectotherms and homeothermsdiffer substantially. The results of recent studies using invitro brainstemspinal cord preparations of adult frogs and premetamorphictadpoles (Rana catesbeiana and Rana pipiens) demonstrate, however,that the mechanisms for rhythm generation and pattern formationdescribed previously for mammals are also key features of therespiratory control system of frogs. These findings thereforesupport the hypothesis that the respiratory control system ishighly conserved amongst air breathing vertebrates, whetherthey breathe continuously or episodically.     

Comparative study of some haematological and biochemical blood parameters in fishes from the Skagerrak

Marine fishes caught in the Skagerrak, 27 different species representing various groups of fishes (Cyclostomi, Holocephali, Elasmobranchii and Teleostei), were examined for the following haematological and biochemical blood parameters: haematocrit, haemoglobin, mean corpuscle haemoglobin concentration, total plasma protein, blood glucose and blood lactate. Interspecies variations as well as variations within some species were observed. The haemoglobin values for all species showed a positive correlation to the corresponding haematocrit values. Relatively low values for haematocrit and haemoglobin were found in cyclostomes, holocephaleans and elasmobranchii compared to the majority of teleosts. Within the teleost group, the haematocrit and haemoglobin levels were positively correlated with the activity of the fish species. The cyclostome Myxine glutimsa L. had a total plasma protein content in the same range as most teleosts, whereas holocephaleans, elasmobranchii and the deep-water teleost Coryphaenoides rupestris Gunnerus showed comparatively low values. Among teleosts some relationship seemed to exist between the total plasma protein level and the activity of the fish species. In addition, a correlation between plasma protein content and levels of blood lipids were noted. Values for blood glucose and blood lactate were found to be lower in cyclostomes, holocephaleans and elasmobranchii than in most teleosts. Higher blood glucose levels were observed in the more active teleost species.     

Modulators of haemocyanin oxygen affinity in the hypoxia- and sulphide-tolerant Baltic isopod Saduria entomon (L.)

Dialysed haemocyanin from the isopod Saduria entomon had a considerably increased oxygen affinity (lower P50) and Bohr factor (-1.71) compared to native haemocyanin (Bohr factor -1.36) indicating that dialysis removes a small molecule size modulating factor decreasing the affinity of native haemolymph. Dialysed haemocyanin had a slightly lower co-operativity (2.42 +/- 0.3) than native haemocyanin (2.9 +/- 0.2). L-Lactate (10 mmol l(-1)) improved oxygen affinity by 1-1.5 torr while urate had no effect. Mg2+ affected affinity in a pH-dependent manner (Bohr-factor increased to -1.67) while Ca2+ had no effect on the Bohr factor but increased affinity with ca 1 torr. Thiosulphate changed the Bohr factor to -1.75 to -1.82, similar to dialysed blood. Co-operativity was in neither case affected. The haemocyanin characteristics of S. entomon are similar to those of crustaceans from hydrothermal vents. These characteristics are probably general for crustaceans that are more or less permanently exposed to sulphide.     

Locomotion, respiratory physiology, and energetics of amphibious and terrestrial crabs

The transition from breathing air to breathing water requires physiological and morphological adaptations. The study of crustaceans in transitional habitats provides important information as to the nature of these adaptations. This article addresses the physiology of air breathing in amphibious and terrestrial crabs and their relative locomotor abilities. Potamonautes warreni is an apparently amphibious freshwater crab from southern Africa, Cardisoma hirtipes is an air-breathing gecarcinid crab with some dependency on freshwater, and Gecarcoidea natalis is an obligate air-breathing gecarcinid endemic to Christmas Island in the Indian Ocean. All three species have well-developed lungs but retain gills and show seasonally different activity patterns that, in the gercarcinids, especially G. natalis, include long-distance breeding migrations. The three species were better at breathing air than water, but P. warreni was the best at breathing water. Cardisoma hirtipes is essentially an obligate air breather and appears to experience facultative hypometabolism during immersion. Cardisoma hirtipes has a haemocyanin with a high affinity for O(2) that facilitates loading from air but makes 30% of the Hc bound O(2) inaccessible. The gecarcinids but not P. warreni show increased diffusion limitation for O(2) over the lung during exercise. Gecarcoidea natalis outperforms C. hirtipes by virtue of a unique haemolymph shunt from the lung into the gills. Paradoxically, it is modifications of the gills for aerial O(2) uptake in G. natalis that allow for relatively greater haemolymph oxygenation. Despite showing decreased arterial-venous DeltaPo(2), P. warreni increased the arterial-venous Delta[O(2)] with no recourse to anaerobiosis during 5 min exercise. In the short term, P. warreni is more adept at walking than C. hirtipes. The breeding migrations of C. hirtipes and G. natalis were completely aerobic, but G. natalis walk farther and probably faster. Seasonal changes in underlying metabolism of G. natalis are strongly implied, including variations in hyperglycaemic hormone, variable basal metabolic rates, and a diel alkalosis present only in migrating crabs. The persistent dependence on water for reproduction is a determining factor in the biology of air-breathing crabs. The annual migrations include costs other than locomotion, for example, burrow construction and intermale competition. Estimates of costs that consider walking alone will underestimate the metabolic and stored fuel requirements for successful reproduction.     

The differential cardio-respiratory responses to ambient hypoxia and systemic hypoxaemia in the South American lungfish, Lepidosiren paradoxa

Lungfishes (Dipnoi) occupy an evolutionary transition between water and air breathing and possess well-developed lungs and reduced gills. The South American species, Lepidosiren paradoxa, is an obligate air-breather and has the lowest aquatic respiration of the three extant genera. To study the relative importance, location and modality of reflexogenic sites sensitive to oxygen in the generation of cardio-respiratory responses, we measured ventilatory responses to changes in ambient oxygen and to reductions in blood oxygen content. Animals were exposed to aquatic and aerial hypoxia, both separately and in combination. While aerial hypoxia elicited brisk ventilatory responses, aquatic hypoxia had no effect, indicating a primary role for internal rather than branchial receptors. Reducing haematocrit and blood oxygen content by approximately 50% did not affect ventilation during normoxia, showing that the specific modality of the internal oxygen sensitive chemoreceptors is blood PO(2) per se and not oxygen concentration. In light of previous studies, it appears that the heart rate responses and the changes in pulmonary ventilation during oxygen shortage are similar in lungfish and tetrapods. Furthermore, the modality of the oxygen receptors controlling these responses is similar to tetrapods. Because the cardio-respiratory responses and the modality of the oxygen receptors differ from typical water-breathing teleosts, it appears that many of the changes in the mechanisms exerting reflex control over cardio-respiratory functions occurred at an early stage in vertebrate evolution.     

A scanning and transmission electron microscopic study of the bat lung

The lungs of two adult species of bat Epomophorus wahlbergi and Miniopterus minor fixed with 2.3% glutaraldehyde were processed for SEM (scanning electron microscope) and TEM (transmission electron microscope) examination by the standard procedures. The bat lung comprised a blood and air conducting zone (consisting of bronchi, bronchioles and large blood vessels), the intermediate zone (made up of alveolar ducts), and the respiratory zone, which consisted of alveoli and blood capillaries. The interalveolar septa comprised basically granular pneumocytes (type II cells), squamous pneumocytes (type I cells), endothelial cells, and, in the interstitium, collagen and elastic fibres with occasional fibrocytes. Blood capillaries were interposed in the interalveolar septa, thus bulging into adjacent alveoli. It was noted that grossly, architecturally and structurally, the bat lung was similar to that of a terrestrial mammal. However, in previous morphometric and physiological studies it has been found that bats have a large lung, a thin pulmonary blood-gas barrier, a large pulmonary capillary blood volume, and high haematocrit and haemoglobin concentration. The bat lung, while retaining the basic mammalian pulmonary design, is well adapted to provide the large amount of oxygen demanded by flight. The avian pulmonary design (the lung-air sac system) is thus not a prerequisite to flight.     

Bimodal breathing in the estuarine crab Chasmagnathus granulatus Dana 1851--physiological and morphological studies

Chasmagnathus granulatus is an estuarine crab which actively moves from subtidal to supratidal areas. To elucidate the possible existence of extrabranchial sites for aerial gas exchange, we measured respiratory and acid-base variables in animals with and without branchial water (controls and experimental crabs, respectively) during air exposure. An histological study of the branchiostegite was also performed. Throughout 4 h of emergence C. granulatus did not suffer venous hypoxia, even without branchial water. The rate of oxygen uptake (M(O(2))) was similar in both groups. The rate of carbon dioxide excretion (M(CO(2))) and the gas exchange ratio (R) significantly decreased during emergence in both groups, with R significantly lower for experimental crabs. Consequently, CO(2) was accumulated in the hemolymph. This variable stabilized after 90 min in control animals, but experimental crabs continued accumulating CO(2). Histological study of the branchiostegites demonstrated the presence of an attenuated and greatly perfused epithelium facing the branchial chamber lumen, with a shortest diffusion distance of 0.5 microm. Simple folds and lobulated projections increase the respiratory surface area. These results suggest that C. granulatus is a bimodal breathing crab, active both in water and air. When emerged, this species extract oxygen directly from air through branchiostegal lungs, but relies on branchial exchange to eliminate carbon dioxide.