A light and electron microscope study of the gills of larval lampreys (Geotria australis) with particular reference to the water-blood pathway.

The gills of ammocoetes of the Southern Hemisphere lamprey Geotria australis have been studied using light and electron microscopy. Emphasis has been placed on describing the structures and vessels involved in gaseous exchange, and on providing quantitative data for the water-blood barrier, including diffusion distance, diffusing capacity and the relative volumes of the component tissues. Although lamprey gills lie inside rather than outside the branchial skeleton as in gnathostomatous fishes. the morphology and ultrastructure of the gill filaments and secondary lamellae of G. australis larvae are very similar to those of teleost fishes. The extensive blood spaces within the secondary lamellae are enclosed by pillar cell bodies and pillar cell flanges which support two layers of epithelial cells. The outer surfaces of the epithelial cells are ridged and covered in a flocculent material which probably represents mucus. Differences were observed in the components of the water-blood barrier at the distal edges and at the surface of the secondary lamellae. At the distal edge, the lining of the marginal channel consisted of an endothelial cell rather than the pillar cell flanges which line the blood spaces of other regions. Based on light micrograph measurements, these differences result in a reduction in the arithmetic mean thickness of the water-blood barrier from 3.62 μm over the pillar cells to 2.22 μm over the marginal channel. Using values for the water-blood barrier obtained from light micrographs, the arithmetic and harmonic mean diffusing capacities were calculated as 1.1046 and 1.7589 ml O₂min/mm Hg/Kg.     

Gill Morphometrics of the Lampreys Lampetra fluviatilis (L.) and Lampetra planeri (Bloch)

Morphometric measurements have been made on various gill components of different stages in the life cycle of the anadromous parasitic lamprey, Lampetra fluviatilis, and its nonparasitic derivative Lampetra planeri. The total gill area, expressed in terms of body weight, of both larval (1462–2717 mm² g^–1) and adult (1402–2337 mm² g^–1) L. fluviatilis are greater than those previously recorded in the rather meagre literature on lamprey gill measurements and are comparable with those found in the most active teleosts. The gills of the two Lampetra species are apparently identical in the larval stages and those of metamorphosing and adult L. planeri are similar to those of metamorphosing L. fluviatilis. Although the pharyngeal arrangement of lampreys differs greatly from that of teleosts, there are many features of the gills indicative of convergence between the two groups. Thus, in a given stage in the life cycle of lampreys, the secondary lamellae on either side of the filaments also alternate, become more widely spaced as the filament length increases and increase in area as the body weight becomes greater. Furthermore, the fractional cumulative increase in secondary lamellae area along a line following the presumed direction of water flow is also represented by a sigmoid curve. While at metamorphosis the pharynx becomes considerably modified to accommodate the change from a unidirectional to a tidal respiratory water flow, the total gill areas of the ammocoete are similar to those of metamorphosing stages which have attained adult characteristics. However, there are clearly differences in some of the components that influence and contribute towards the total gill area. Thus, in terms of body weight, the number and total length of the filaments and the total number of secondary lamellae, together with the number of secondary lamellae found on a given distance of filament, are greater in late metamorphosing stages, while the reverse is true for the average bilateral area of the secondary lamellae which is considerably greater in ammocoetes.     

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.     

Structural organization of the gills in pipefish (Teleostei,Syngnathidae)

Summary The morphology and structural features of the gills of the two Western Baltic pipefish Nerophis ophidion and Syngnathus rostellatus were investigated using scanning electron microscopy. The general anatomy of the gills complies with the general pattern in fish. Several adaptations though, show the highly specialized nature of pipefish gills. The filaments are extremely short, few in number and carry only a few lamellae due to the limited space in the branchial cavity. The lamellae have a widely projecting form yet still have a small area in comparison to other fish. Gill irrigation is performed by a specialized pumping mechanism which forces respiratory water through the small but densely packed gill sieve. Although both species live in the same habitat and belong to the same family, differences in gill morphology were found and are related to different lifestyles. S. rostellatus is the more active species and therefore has more filaments per gill arch, more lamellae per filament, wider projecting lamellae and a more extreme utilisation of available space in the gill cavity through a very densely packed gill sieve. N. ophidion has a stationary mode of life and therefore has a less extreme gill anatomy.     

Early development and microhabitat of the monogenean Horricauda rhinobatidis, with observations on the related Troglocephalus rhinobatidis, from Rhinobatos batillum from Queensland,Australia

The small monocotylid monogenean Horricauda rhinobatidis is abundant on the gills of its host Rhinobatos batillum, whereas the larger, related monogenean Troglocephalus rhinobatidis is comparatively uncommon. Young specimens of Horricauda live between the host's secondary gill lamellae. Post-oncomiracidia have 14 marginal booklets but as the larvae develop these are supplemented first by a pair of hamuli and then by muscular ventral loculi followed by six forwardly-directed, dorsal spines. By impaling secondary gill lamellae these spines may serve to prevent parasites from being dislodged by gill ventilating currents. Before reaching sexual maturity the parasites leave the secondary gill lamellae and establish themselves in the septal canals. It is uncommon to find more than one adult specimen of Horricauda in each septal canal. The significance of this in relation to sperm exchange is discussed. Like Troglocephalus, Horricauda has eight head sacs, and the suggestion is made that these sacs may play a part in feeding.     

Morphology and ultrastructure of the gills of terrestrial crabs (Crustacea,Gecarcinidae and Grapsidae): Adaptations for air-breathing

Summary The terrestrial crabsGeograpsus grayi, Geograpsus crinipes, Cardisoma hirtipes andGecarcoidea natalis have a reduced number of gills and show a reduced planar gill surface (SA) compared to aquatic species. Gill lamellae are stiffened and thickened (increasing blood/gas (BG) diffusion distances) and nodules maintain wide spacing between lamellae. Haemolymph is directed through the gill lamellae by rows of pillar cells and in the afferent region an intralamellar septum splits the haemolymph into two parallel networks. Gaps in the lines of pillar cells allow movement of haemolymph between adjacent channels. The afferent vessel distributes haemolymph to the lamella via a number of direct channels including the marginal canal and in large gills with the aid of a long, forked sinus which supplies the ventral and central regions of the lamellae. The marginal canal functions in both distribution and collection of haemolymph; the role varies with species. Potential flow-control sites were identified at the junctions between afferent and efferent areas and where the efferent channels enter the efferent branchial vessel. Each gill receives a branch from the sternal artery which supplies all the lamellae. Transport epithelia is the principal cell type in the gills of all species examined though its location varies between species, either being confined to certain gills or specific parts of the lamellae.The gill lamellae of air-breathing crabs are clearly modified to breathe air (stiffening and presence of nodules), though the overall contribution of the gills to gas exchange has been reduced (smaller SA and longer BG diffusion distances). The role of the gills in air-breathing crabs thus appears to have switched from one of an efficient aquatic gas-exchanger (thin with large surface area) and transport tissue, to one that is predominantly set up for ion-regulation.Abbreviations a afferent branchial vessel - ac afferent channels - art arteriole - ass artifactual subcuticular space - bl basal lamina - c cuticle - col collagen - ct connective tissue - e efferent branchial vessel - ec efferent channels - epi epithelium - f folds - g Glycogen - h haemolymph - hc haemocyte - is intralamellar septum - m marginal canal - mi mitochondria - mt microtubules - n nucleus - p pillar cell - s shaft of efferent vessel - sd septate desmosome     

The gill of fish larvae. Is it primarily a respiratory or an ionoregulatory structure?

Based on surface area and chloride cell number, rainbow trout Oncorhynchus mykiss gills appear initially to be more important in terms of ion balance than gas exchange. Chloride cells appear on the gill 3–6 days before hatch at 10°C. This is about 9 days before secondary lamellae, the definitive adult gas exchange structure, begin to form. At hatch, 22% of all chloride cells but only 7% of total surface area are located on the gill. This discrepancy gradually diminishes but even at complete yolk absorption the gill still seems to be about twice as important in terms of ion balance (75% of chloride cells) as gas exchange (37% of total surface area represented by gill filaments and lamellae combined). Surface area measurements and chloride cell counts reported in the literature for larvae of other species show a similar pattern suggesting that this may be a general phenomenon. If true, the implications are profound not only for developmental physiologists but also for those interested in the evolutionary history of gills and their function in adult fish.     

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.     

Changes in gill morphology of Oreochromis mossambicus subjected to heat stress

Synopsis The cichlid fish, Oreochromis mossambicus, was acclimated to 25°C for a 14 day period and then subjected to elevated temperatures of 30, 35, 40 and 45° C. Gill epithelia was progressively damaged from 30–40° C; gills from fish tested at 45°C showed less damage than those tested at 40°C. This is presumably due to the shorter exposure (survival) time at 45°C. Shrinkage of the secondary lamellae and the collapse of pillar cells supports the hypothesis that hypoxia at elevated temperatures is partially a function of changes in gill morphology.     

High environmental temperature and low pH stress alter the gill phenotypic plasticity of Hoven's carp Leptobarbus hoevenii

Climate warming and low pH environment are known to negatively impact all levels of aquatic organism from cellular to organism and population levels. For ammonotelic freshwater species, any abiotic factor fluctuation will cause disturbance to the fish, specifically at the gills which act as a multifunctional organ to support all biological processes. Therefore, this study was designed to investigate the effect of temperature (28 vs. 32°C) and pH (7.0 vs. 5.0) stress on the gill plasticity of Hoven's carp after 20 days of continuous exposure. The results demonstrated that high temperature and low pH caused severe changes on the primary and secondary lamellae as well as the cells within lamellae. An increasing trend of the proportion available for gas exchange was noticed at high temperature in both pH exposures, which resulted from a reduction of the primary lamellae width with elongated and thinner secondary lamellae compared to fishes at ambient temperature. Following exposure to high temperature and acidic pH, Hoven's carp experienced gill modifications including aneurysm, oedema, hypertrophy, curling of secondary lamellae, epithelial lifting, hyperplasia and lamellae fusion. These modifications are indicators of the coping mechanism of Hoven's carp to the changing environment in order to survive.     

The effect of input pressure and flow on the pattern and resistance to flow in the isolated perfused gill of a teleost fish

Summary When an isolated gill arch of the marine teleost,Ophiodon elongatus, was perfused under conditions which mimic those present in the intact animal, only two thirds of the gill lamellae were perfused. An increase in either input (afferent) pressure and flow or input pulse pressure caused an increase in the number of lamellae perfused as well as altering the distribution of the efferent outflow between the efferent artery and the venolymphatic drainage of the gill. The gill is compliant and an increase in efferent pressure reduced gill resistance to flow without altering the number of lamellae perfused. In these experiments there was no simple relationship between the number of lamellae perfused and gill resistance.These observations are of importance in the interpretation of results from pharmacological and ion exchange studies of isolated gills as well as indicating how cardiovascular changes could bring about alterations in gill blood flow in the intact fish.     

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.     

Interspecific variations in the surface ultrastructure of the gills of freshwater mullets

SEM studies were made on the gills of freshwater mullets,Rhinomugil corsula andSicamugil cascasia, to correlate surface ultrastructure of various gill units with their probable functions. Two types of lamellated gill rakers of the former fish are suited for plankton feeding and the short, stumpy and transversely beaded gill rakers of the latter reflect the varied food and feeding habit of the fish.R. corsula has numerous mucous glands on the epithelium covering the gill arch and gill filaments,S. cascasia has fewer. In accordance with the differences in the density and distribution of the mucous glands, the microridged epithelial cells also show variations in their architectural plan. In both species the epithelium of the secondary lamellae is smooth, probably an adaptation for better gaseous exchange.     

Immunohistological classification of ionocytes in the external gills of larval Japanese black salamander,Hynobius nigrescens Stejneger

In this cytological and immunohistological study, we clarified the localization of the membrane transporters Na⁺, K⁺‐ATPase (NKA), vacuolar‐type H⁺‐ATPase (VHA), and epithelial sodium channel (ENaC) and distinguished ionocyte subtypes in the gill of the Japanese salamander (Hynobius nigrescens). In larvae (IY stages 43–65), NKA immunoreactivity was observed on the basolateral plasma membrane in more than 60% cells and less than 20% cells in the primary filaments and secondary lamellae of the external gills, respectively. VHA immunoreactivity was observed on the apical membrane of some epithelial cells in the secondary lamellae of the external gills. High ENaCα immunoreactivity was widely observed on the apical cell membrane of a population of squamous cells, presumably pavement cells (PVCs), and mitochondria‐rich cells (MRCs), in the primary filaments and secondary lamellae of the external gills. Using double immunofluorescence microscopy, epithelial cell types involved in ionic regulation were characterized and divided into three ionocyte types: NKA‐, NKA‐ and ENaC‐, and VHA‐positive cells. VHA‐immunoreactive cells as well as NKA‐positive cells were observed during IY stages 43–65 of the salamander larvae. During late stages of metamorphosis, NKA, VHA, and ENaCα immunoreactivities in the external gills decreased and finally disappeared during the completion of metamorphosis (IY stage 68). PVCs and MRCs in the external gills are probably involved in acid–base balance regulation and osmoregulation in urodele amphibian larvae. The results are discussed in relation to the ionocytes previously reported in fish gills and the frog skin epithelium. J. Morphol., 2011. © 2011Wiley‐Liss, Inc.     

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.     

The architecture of the gill surface of the catfish, Heteropneutes fossilis (Bloch): SEM study.

A scanning electron microscopical examination of the gills of H. fossilis is described. The surface architecture of gill filaments and secondary gill lamellae showed the presence of various features such as indentations, micropits and crevices. The possible functions of these morphological adaptations are discussed in relation to physiology of the gill.     

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.     

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     

First observations of histopathological effects of 2,4,6-trinitrotoluene (TNT) in gills of European eel <Emphasis Type="Italic">Anguilla anguilla</Emphasis> (Linnaeus, 1758)

The aim of the present study was to investigate the effects on gill morphology of the explosive 2,4,6-trinitrotoluene (TNT) in a model fish, the European eel, Anguilla anguilla (Linnaeus, 1758) to assess potential detrimental effects in marine fish due to its presence in dumping areas. Juvenile specimens of A. anguilla were exposed in vivo for 6 and 24 h to 0.5, 1 and 2.5 mg/l nominal concentrations of TNT using dimethyl sulfoxide (0.1‰) as solvent carrier. Histological analysis of gills indicated that TNT induced several structural lesions. After 6h of exposure at 0.5 and 1 mg/l TNT, oedema of some secondary lamellae was evident: this change at the highest dose of 2.5 mg/l led to epithelial lifting and detachment from the endothelium and rupture of the branchial membrane and vascular congestion. After 24 h at 0.5 mg/l, increased oedema in secondary lamellae, extensive epithelial detachment and vascular congestion and dilation of lamellar capillaries and pooling of blood at 1 mg/l were observed. At 2.5 mg/l, epithelial hyperplasia, fusion of adjacent lamellae, obliteration of interlamellar spaces by means of tight junctions were also observed. Moreover, chloride cells proliferated along secondary lamellae, and mucus hypersecretion was evident. The overall results clearly indicate that gills are sensitive targets of TNT.     

Fish gill morphology: inside out

In this short review of fish gill morphology we cover some basic gross anatomy as well as in some more detail the microscopic anatomy of the branchial epithelia from representatives of the major extant groups of fishes (Agnathans, Elasmobranchs, and Teleosts). The agnathan hagfishes have primitive gill pouches, while the lampreys have arch-like gills similar to the higher fishes. In the lampreys and elasmobranchs, the gill filaments are supported by a complete interbranchial septum and water exits via external branchial slits or pores. In contrast, the teleost interbranchial septum is much reduced, leaving the ends of the filaments unattached, and the multiple gill openings are replaced by the single caudal opening of the operculum. The basic functional unit of the gill is the filament, which supports rows of plate-like lamellae. The lamellae are designed for gas exchange with a large surface area and a thin epithelium surrounding a well-vascularized core of pillar cell capillaries. The lamellae are positioned for the blood flow to be counter-current to the water flow over the gills. Despite marked differences in the gross anatomy of the gill among the various groups, the cellular constituents of the epithelium are remarkably similar. The lamellar gas-exchange surface is covered by squamous pavement cells, while large, mitochondria-rich, ionocytes and mucocytes are found in greatest frequency in the filament epithelium. Demands for ionoregulation can often upset this balance. There has been much study of the structure and function of the branchial mitochondria-rich cells. These cells are generally characterized by a high mitochondrial density and an amplification of the basolateral membrane through folding or the presence of an intracellular tubular system. Morphological subtypes of MRCs as well as some methods of MRC detection are discussed.