Morphology of the gills of larval and parasitic adult sea lamprey,Petromyzon marinus L.

The general morphology of the gills is similar in larval (ammocoetes) and parasitic adult sea lampreys, Petromyzon marinus, despite different methods of ventilation necessitated by their feeding habits. The gill lamellae are supported by randomly-distributed pillar cells which enclose blood spaces and collagen columns. The distribution of these cells in lampreys is different from that of higher fishes and it may be inefficient for respiratory exchange. The presence of cytoplasmic microfilaments suggests that these cells have the ability to reduce the lamellar blood spaces through contraction. Marginal channels at the tips of the lamellae are lined only by endothelial cells. The thickness of the water-blood pathway in lampreys falls within the range described for higher fishes, with the most efficient gas exchange likely occurring at the lamellar tips where only a single layer of epithelial cells is present. The abrupt increase in height of the epithelium near the lamellar bases in adults, compared to the gradual transition in height along the lamellae in ammocoetes, is perhaps reflective of higher oxygen requirements during the parasitic stage. The consistent appearance of wide, lateral intercellular spaces within the respiratory epithelium of lampreys indicates possible involvement of these spaces in transport. Mucous secretion appears to be an important function of the superficial platelet cells in ammocoetes. “Mitochondria-rich” and “mitochondria-poor” superficial cells are observed in both ammocoetes and adults, with the mitochondria-rich cells more prevalent toward the lamellar bases. The possibility that at least some of these cells may be involved in absorption is discussed. Mitochondria-rich cells in the interlamellar region are morphologically different in ammocoetes and adults but all possess an abundance of smooth endoplasmic reticulum and hence resemble “chloride cells” of higher fishes. The similarity of these cells in the parasitic adult lamprey to chloride cells of marine fishes may reflect the potential of the adult lamprey to osmoregulate in salt water. A scarcity of these cells in ammocoetes and their resemblance to chloride cells in freshwater fishes may reflect the restriction of larval lampreys to a freshwater habitat.     

Fine structure of the gills of some Indian air-breathing fishes

A detailed account is given of the structure of the gills of Clarias batrachus, Heteropneustes (= Saccobranchus) fossilis, Channa punctata, Monopterus (= Amphipnous) cuchia and Boleophthalmus boddaerti, based upon light and electron microscopy. In all five species the basic organization into primary and secondary lamellae is apparent but the latter are very much more modified in Monopterus. Three main layers separate the water and blood on the surface of the secondary lamellae. The outer epithelium is usually two layered but may be multilayered close to the origin of the secondary lamellae from the gill filament. The basement membrane is relatively thin and a middle dense layer containing collagen fibrils separates two clear layers. The pillar cells, so characteristic of secondary lamellae, are present in all except Monopterus and flanges from these cells surround the blood channels with the exception of the marginal channels. The latter are lined by endothelial cells which line all the blood channels of Monopterus. The overall thickness of the three layers comprising the water/blood barrier ranges from 1.5 to 13 microns. A number of modifications to this basic organization can be related to the degree of dependence of the different species on air-breathing. Boleophthalmus is the only species commonly found in brackish water and its secondary lamellae have well developed lymphoid spaces between two layers of the epithelium. Special densely-stained regions of the pillar cell flanges were also present in this fish and may have a supporting function.     

Fine structure of the respiratory organs of the Climbing perch, Anabas testudineus (Pisces: Anabantidae)

An electron microscopic study has been made of the three respiratory organs of climbing perch. The gill structure is similar to that of the other telcosts but the thickness of the water/blood barrier is much greater, being as great as 20 μm in some specimens. The increased thickness is due to a multilayered epithelium which is thinner (3.5–7 μm) over the marginal channel of the secondary lamellae. The other two main layers, basement membrane and pillar cell flange, are relatively thin (about 1 μm).
The pillar cells have a typical structure, but in certain regions they are contiguous with one another and line well-defined blood channels. Some of the columns of basement membrane material in such regions may be common to adjacent pillar cells.
The air-breathing organs are (a) the lining of the suprabranchial chambers , and (b) the labyrinthine plates attached to the dorsal region of branchial arches. Electron microscopy showed that their structure is well adapted for gas exchange, the air/blood barriers being only 0.12–0.3 μm, comprising an epithelial layer, basement membrane, and thin capillary endothelium. The many parallel blood channels of the respiratory islets of both organs are separated by pillar-like structures which differ from the pillar cells of the secondary lamellae. Thus the hypothesis that the air-breathing organs represent modified gills is not supported by this study.
The fine structure of the non-respiratory region of the air-breathing organs is similar to that of the skin, and includes chemoreceptor-like cells. Evidence concerning the possible homology of pillar cells with plain muscle cells is discussed.     

Nature of the air-breathing organs of the Indian fishes Channa, Amphipnous, Clarias and Saccobranchus as shown by electron microscopy

The structure of the air-breathing organs of the Indian fishes Channa punctatus, Channa Striatus, Amphipnous cuchia, Clarias batrachus and Saccobranchus fossilis has been investigated using electron microscopy. In all species the barrier separating the air from the blood consists of three main layers (epithelium, basal lamina and endothelium). The total thickness ranging from 0.78 μm in C. punctatus 1.6 μm in S. fossilis.
In Clarias and Saccobranchus the presence of pillar cells characteristic of gill secondary lamellae confirms evidence for the origin of these organs by modification of a typical gill structure.
In Amphipnous and two species of Channa , however, the evidence suggests that the accessory organs represent modified gills. The presence of valve-like structures between the afferent and efferent blood spaces of the vascular papillae gave the appearance of pillar cells under the light microscope.
The structure of these organs is correlated with physiological studies on the degree of their importance in the life of the animal and the degree of gill development     

THE STRUCTURE OF FISH GILLS IN RELATION TO THEIR RESPIRATORY FUNCTION

1. The general structure of the gills of different fishes is compared and it is concluded that, though essentially the same, there are certain differences by which they can be recognized. Possible ways in which they may have evolved from one another are considered. 2. A detailed account is given of the structure of the secondary lamellae, where gaseous exchange takes place, and it is shown that two epithelial sheets are separated by a vascular axis mainly composed of pillar cells overlain by a basement membrane on each side. Blood pathways through the gills are discussed in relation to their respiratory function. 3. The embryonic development of gills is described and evidence regarding homo-logies of different structures, particularly the pillar cells, is reviewed. 4. The gills of fish having different modes of life show variations in (a) the number of arches, (b) the number and length of the gill filaments, and (c) the size and frequency of the secondary lamellae. Ways in which measurements of gill area may be carried out and some of the complications involved are reviewed and a summary given of measurements made for a wide variety of species. Measurements of the thickness of the water-blood barrier are also discussed; the more active fish generally have thinner water-blood barriers and larger gill areas. 5. The different mechanisms of gill ventilation are summarized and characteristics of gill resistance in elasmobranchs and teleosts are compared. Gas exchange is discussed in relation to available techniques and the current terminology and symbols, and to indicate the value of analogies between gill exchangers and systems studied by engineers. 6. It is outlined how studies of the functioning of gills during coughing, parasitic infection, and in polluted waters add to knowledge of their role in respiration.     

Intercellular junctions in the water-blood barrier of the gill lamella in the adult lamprey (Geotria australis,Lampetra fluviatilis)

Thin sections and freeze-fracture replicas of the water-blood barrier in the gill lamellae of adult lampreys (Geotria australis, Lampetra fluviatilis) demonstrate that the occluding junctions between epithelial pavement cells differ markedly from those between endothelial pillar cells in the structure and arrangement of their strands. The zonulae occludentes between pavement cells typically consist of complex networks of 4–6 strands, the mean number of which undergoes a small but significant decline when the animal is acclimated to seawater. In comparison, the occluding junctions between pillar cells are less elaborate and may represent maculae or fasciae, rather than zonulae occludentes. They do not apparently undergo a change when the animal enters saltwater. The results indicate that the main part of the paracellular diffusion barrier to proteins and ions is located in the epithelium rather than the endothelium. Communicating (gap) junctions are present between adjacent pavement cells, between pavement and basal cells and between pillar cells. These findings suggest that the epithelial cells and the pillar cells in the water-blood barrier of lampreys both form functional syncytia. The results are discussed in the context of ion-transporting epithelia in other aquatic vertebrates.This paper is dedicated to Professor H. Leonhardt on the occasion of his 75^th birthday     

A morphological study on posterior gills of the mangrove crab Ucides cordatus

Morphological and histological studies on posterior gills of the mangrove crab Ucides cordatus showed that the 5th gill (of 7) has a larger surface area and a greater number of lamellae compared to the 6th gill. Regular separation of gill lamellae, important when the gill is in air, is maintained by enlargements of the marginal canals. Conical, spine-like structures along the efferent vessel of both 5th and 6th gills were also observed. In addition, pillar cells, a discontinuous lamellar septum and a hypobranchial artery were observed. The presence of valve-like structures near the efferent vessel was also indicated. These structures, together with the pillar cells, may have a role in directing the hemolymph flow towards certain gills during particular physiological states. Localization of osmoregulatory epithelia in the lamellae of both gills was inferred from dimethylaminostyrylethylpyridiniumiodine staining. Apparently gills 5 and 6 have osmoregulatory epithelial cell patches of similar area, corresponding to 43% and 38% of the total lamellae area, respectively. However, their localization is quite different. Gill number 5 osmoregulatory patches seem to be restricted to the afferent region of the lamella whereas in gill number 6, they are more dispersed over the entire lamella. These differences may be related to the particular functional characteristics of these gills.     

Gross and fine structure of the pseudo branch of the climbing perch, Anabas testudineus (Bloch)

The structure of the glandular pseudobranch of the air-breathing fish Anabas testudineus is described on the basis of light and electron microscopy. It is shown that the pseudobranch has the same basic structure as a typical gill, although in this case it is not freely exposed to the water.
Individual secondary lamellae can be recognized in which well-defined, but narrow, blood channels are present between typical pillar cells. The epithelial layer is mainly represented by enlarged mitochondria-rich cells which constitute a large proportion of the whole organ. Mitochondria-rich cells contain an abundant endoplasmic reticulum which is in close contact with the mitochondria and becomes concentrated near the vascular borders of the cell and opens directly into the basement lamina.
The presence of numerous pinocytotic vesicles in the enlarged pillar cell flanges may transfer material to the blood channels. The precise nature and role of any materials released in this way remains to be investigated.     

Direct and circulatory paths of Permethrin (NRDC-143) causing histopathological changes in the gills of rainbow trout, Salmo gairdneri Richardson

Rainbow trout were exposed to sublethal concentrations of Permethrin in water (0.09, 0.18 and 0.35 μg l⁻¹) and in food (85, 180 and 350 μg kg⁻¹ dry diet) in 20–0 day experiments. Histological changes in the gills included epithelial separation or necrosis, mucus cell hyperplasia, clubbing of epithelial cells or hyperplasia and fusion of adjacent secondary lamellae. These changes were noted in all fish, suggesting that the pesticide or its metabolites reached the gills not only directly through the water, but also indirectly via the circulation.     

Gill ultrastructure of the Pacific hagfish Eptatretus stouti

At the gross anatomical level, hagfish gills show unusual features not seen in any other fish gills. Our study was undertaken to determine if peculiarities also characterize the microscopic anatomy and ultrastructure of hagfish gills. To the contrary, branchial respiratory lamellae of Pacific hagfish were found to resemble the lamellae of lampreys, elasmobranchs, and teleosts, often down to the finest subcellular details. As in other fish, hagfish lamellae are lined by epithelium containing pavement cells with organelles indicative of a secretory function, basal cells showing undifferentiated cell features, and branchial ionocytes. The ionocytes are identical to chloride cells of teleosts in cytostructure, distribution, and abundance. There are pillar and marginal capillaries in hagfish gill lamellae. Pillar cells contain bundles of 5-nm microfilaments, and they associate with collagen columns as in other fish. Hagfish pillar cells do exhibit odd features, however: They cluster (groups of up to nine were seen), and their extracellular collagen columns are rarer than in other fish gills (averaging only two columns per three pillar cells). Other special features of hagfish gills are the following: lipid droplets and smooth endoplasmic reticulum are well developed in all cell types; pavement cells secrete a lipomucous product (stains with periodic acid-Schiff, Alcian blue, and Sudan black B); and goblet cells are absent. The presence of "chloride cells" in hagfish is puzzling, as hagfish body fluids are iso-osmotic to seawater and there is no need to osmoregulate for sodium chloride; the ionocytes contain carbonic anhydrase, suggesting a function in acid/base regulation.     

Oxygen uptake during early life in the fresh water fish, Esomus danricus (Ham) (Pisces, Cypriniformes)

O2 uptake in Esomus danricus has been determined in relation to body weight, length and thickness of the water-blood diffusion barrier at 27-28 degrees C temperature. Total O2 consumption in larvae was 1311 ml/kg/h but decreased significantly in juvenile fishes (720 ml/kg/h). The increase in the thickness of water-blood diffusion barrier at the secondary gill lamellae of the fish was found to be an important factor for the decrease in VO2. Logarithmic analyses of data for O2 uptake in relation to body weight gave a slope of 0.8865 for larvae and 0.5053 for juveniles. The exponent values of O2 uptake against diffusion barrier for larvae and juveniles were 1.7383 and 2.0956, respectively. The results obtained indicated that fish have an extra device which helps in extracting about 24% of the total VO2 required for the fulfilment of the metabolic oxygen demand of the body.     

A comparative study of the ultrastructure of the water-blood pathway in the secondary lamellae of teleost and elasmobranch fishes — benthic forms

Summary The ultrastructure of the secondary lamellae has been examined in four species of free-swimming elasmobranchs, two species of Raia and five species of bony flatfishes. Microvilli which are present on the outer epithelial surface vary in form and size. It is suggested that a possible function of the microvilli is to anchor a surface mucus covering whose possible functions are discussed. Vesicles are found immediately beneath the microvilli in elasmobranchs but not in teleosts. Chloride and mucous cells are present on the secondary lamellae of all species and often have microvilli of greater length than on the other cells of the epithelium. Micropinocytotic vesicles are found on both sides of the basement membrane. The number of columns enclosed in the pillar cells varied from 5–11.The water-blood pathway showed variations in thickness not only between different species but also in different parts of individual secondary lamellae. A general trend in the mean total thickness was found, being greater in the swimming elasmobranchs (10.22 ) than in the rays (5.47 ) and bony flatfish (3.59 ). This trend is also seen in the greater length of microvilli and thickness of basement membrane in the elasmobranchs than in the teleosts.     

Gill infection of Leporinus macrocephalus Garavello & Britski, 1988 (Osteichthyes: Anostomidae) by Henneguya leporinicola n. sp. (Myxozoa: Myxobolidae). Description, histopathology and treatment

Piau?us (Leporinus macrocephalus), were raised in 300 m2 ponds (density of 10 fish/m2) presenting asphyxia signals and daily mortality of 27 fishes. Specimens with 8-cm total body length, were collected for necropsy. Mucus of body surface and pieces of organs were collected and examined microscopically, in wet mounts, stained or in histological sections. The smears examination showed the presence of several spores in the secondary lamellae of the gill filaments, identified as Henneguya leporinicola n.sp (Myxozoa: Myxobolidae). Histopathological study showed epithelial hyperplasia and fulfilling of the spaces between the secondary lamellae, congestion and telangiectasia sinusoidal. It was also observed hyperplasia of the goblet cells and several cysts of parasite with 70.3 microns diameter. Such cysts were situated among the secondary lamellae, covered or not by the hyperplasic epithelium. With this diagnostic, three applications of formalin solution 10 ml/m3 were carried out. Fifteen days after that, fish were examined again to ascertain whether the treatment was efficient on disease caused by the protozoa. The tissue alterations present in the gills after the treatment were just a moderate sinusoidal congestion and a slight epithelial hyperplasia on the base of the secondary lamellae.     

Cadmium-induced changes in gill morphology of zebrafish, Brachydanio rerio (Hamilton–Buchanan), and rainbow trout, Salmo gairdneri Richardson

Freshwater zebrafish and brackish water rainbow trout were exposed to different concentrations of cadmium for up to 6 weeks. The gill morphology was examined by light and electron microscopy, and a morphometric analysis was performed. The morphometric study of the secondary lamellae revealed an increase in the portion exterior to the basal lamina, resulting in an increased diffusion distance, after exposure to cadmium concentrations of 10μg l⁻¹ and above. In both species this was due to an increase in volume of the non-tissue spaces of the secondary lamellar epithelium. Furthermore, the water space between neighbouring secondary lamellae was clearly reduced. Morphological examination revealed some gross alterations compared with control fish. These initially consisted in curling of the secondary lamellae and finally resulted in local teleangiectasia. Partial lifting of the secondary lamellar epithelium from the pillar cells resulted in large non-tissue spaces which were invaded by leucocytes. The first sign of degeneration was observed in the chloride cells, which were characterized by a dispersed cytoplasm and a smooth apical plasma membrane.     

Branchial gas transfer models

Gas transfer in fish gills is simulated by a simple counter-current model, with ventilation, water-blood transfer and blood flow characterized by conductances. The ventilation and perfusion conductances are products of flow rate and effective solubility. The diffusion conductance of water-blood transfer (diffusing capacity) is considered to depend on diffusion properties of both the water-blood tissue barrier and of interlamellar water. In the gills of the dogfish Scyliorhinus stellaris, more than half of the total resistance to O2 diffusion was located into interlamellar water. Complicating factors like water shunt, blood shunt, ventilation-perfusion maldistribution, pulsatile flow, diffusion in blood and reaction of O2 with hemoglobin may reduce the O2 transfer efficacy predicted by the simple model. In S. stellaris, the effect of such complicating factors appeared to be minor in most conditions, but in other species and/or conditions, more complex models might be required.     

Ultrastructure of the secondary gill lamella of the icefish, Chaenocephalus aceratus

The basic structure of the secondary lamella is similar to that found in most fishes, but the water/blood barrier is relatively thick (c. 8 pm) of which about 90% is epithelium. The marginal channel is larger than channels between pillar cells which appear to have contracted.     

Gill morphology in the zebrafish, Brachydanio rerio (Hamilton- Buchanan)

A light and electron microscopic study of the gills of the zebrafish, Brachydanio rerio , were made to serve as a morphological basis for future investigations. It was found that for fixation of B. rerio gills, a mixture of 1·5% gluturaldehyde and 1·5% paraformaldehyde gives a mucus-free surface. Morphometric measurements of structural components of the gill secondary lamellae were made. Observations at SEM were correlated with those made at TEM. The different cell types in the branchial epithelium were characterized. Chloride cells were mainly located in the interlamellar regions and on the afferent side of the primary lamellae. Two morphologically different chloride cells were seen. The first type communicates with the external environment through a reservoir-like lumen, which is normally absent in freshwater fishes. The second type of chloride cell has more direct contact with the ambient water, resembling chloride cells from other freshwater fishes. Another cell type with features similar to those of the rodlet cell was frequently observed. This cell is interposed between other types of cells in the epithelium, and sometimes junctional complexes were present between the rodlet cell and surrounding cells.     

Ultrastructural study of the early development and localization of Loma salmonae in the gills of experimentally infected rainbow trout

The early ultrastructural stages of Loma salmonae were studied in the gills of experimentally infected rainbow trout. No parasitic stages were identified during the first 2 wk of the infection. By week 3 postexposure (PE), uninucleate and binucleate meronts were recognized within host cells (no xenomas) associated with the capillary channels of secondary lamellae and lamellar arteries. An inflammatory reaction was absent. In secondary lamellae, infected cells were isolated from the capillary lumen, and some were recognized as pillar cells. In lamellar arteries, infected cells were localized beneath the endothelium and not in the lumen. Inflammatory reaction and destruction of parasites inside blood cells in the lumen of secondary lamellae were observed by week 4 PE. Three hypotheses, i.e., isolation, internalization, and evasion, are proposed to explain the localization of the infected cells in the gills. It is concluded that meronts are the earliest parasitic stage observed by week 3 PE, pillar cells are secondarily infected by phagocytosis of infected cells in the blood, endothelial cells of gills are not infected, and inflammatory response to the parasite starts by week 4 PE.     

Mineral status of embryos of domestic fowl following exposure in vivo to the carbonic anhydrase inhibitor acetazolamide

Na+/H+ exchangers are integral membrane proteins that exchange Na+ and H+ across cell membranes. The Na+/H+ exchangers 2 and 3 are epithelial isoforms in mammals and contribute to acid-base homeostasis. The gills of fishes, including elasmobranchs, are also associated with acid/base balance, and are probably the primary acid/base regulatory organ. This study examines the presence of Na+/H+ exchangers 2 and 3 using immunohistochemistry and immunoblotting in the gills of four species of elasmobranchs, the banjo ray (Trygonorrhina fasciata), southern eagle ray (Myliobatis australis), the gummy shark (Mustelus antarcticus) and the Australian angel shark (Squatina australis) using heterologous antibodies. Na+/H+ exchanger 2-like immunoreactivity was observed in the gills of the banjo ray, eagle ray and angel shark. In the banjo and eagle rays, this Na+/H+ exchanger-like immunoreactivity co-localised with immunoreactivity to Na+ /K+ -ATPase, a marker for the mitochondrial-rich cells of fishes. Na+/H+ exchanger 3-like immunoreactivity was only observed in the gills of the angel and gummy sharks, some Na+/H+ exchanger 3-like cells also showed Na+ /K+ -ATPase immunoreactivity. However, immunoblotting of banjo and eagle ray gill membranes demonstrated Na+/H+ exchanger 3-like immunoreactivity, which was not consistent with the immunohistochemical results. These data demonstrate the presence of epithelial Na+/H+ exchangers 2 and 3 in the gills of elasmobranchs and a link with acid/base regulation is suggested.     

Electron microscopic studies of the gill epithelium of the amphibian teleost Periophthalmus vulgaris]

The gill epithelium of the airdwelling fish Periophthalmus vulgaris has been studied with the electron microscope. The following celltypes can be distinguished: flat covering epithelial cells, chloride cells, mucous cells, basal cells, various leucocytes as well as a specific granule containing cell which is possibly an epithelial cell. The covering epithelial cells exhibit a relatively smooth apical surface and contain in their apical half densely packed microfilaments, pinocytotic vesicles are rare. These characteristics are not to be found in water dwelling fish and possibly represent adaptations to the air containing surroundings. In the chloride cells are numerous, especially in the basal halves of the secondary lamellae. The distal parts of the secondary lamellae the barrier for the respiratory gases measures about 0,9 micrometer. The basal cells are ribosome rich replacement cells. Two types of mucous cells occur. Individual intraepithelial nerve fibres have been observed.