Gill sphaerosporosis in goldfish (Carassius auratus)

Infections caused by a Sphaerospora sp. resembling S. chinensis are reported for the first time in goldfish (Carassius auratus) from North America. The myxosporean was found in the respiratory epithelium of the gill of pond-reared fish. Spores from stained tissue sections were spherical with an equal mean length and width of 6.3 microns. Spore valves were thickened at the suture which lies in a plane perpendicular to two prominent pyriform polar capsules. The polar capsules were 4.0 x 2.8 microns in length and width. Both monosporous and disporous development within a surrounding "pseudoplasmodium" was detected. Infections caused moderate hyperplasia and occasional necrosis of the respiratory epithelial cells of the gill.     

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.     

Some functional and morphological adaptations in the gills of dipnoi

Summary The gill epithelium of the Neoceratodus forsteri contains ce0lls which produce generations of dark vesicles. These vesicles move in waves towards the surface of the cells, break the cellular membranes and pour their content onto the cellular surfaces. This surface coat consists of threads and granules. The breaking of cell membranes by the densely situated vesicles leads to formation of processes, which join frequently to form a surface net: conversely the cytoplasm of the epithelial cells forms large invaginations penetrated by fluid. The reduction of cytoplasm and the mechanical loss of the cytoplasmic processes leads finally to the destruction of the cell.A small number of granulated cells with long processes extending between other cells have also been observed. The border between the gill epithelium and the connective tissue is characterised by the presence of bundles of thin fibers penetrating the epithelium.The above findings are discussed in relation to the differentiation of the epithelial cells, the facilitation of respiration in poorly oxygenated waters, the protection of gill surfaces against abrasion and with regard to the survival of the lung fish in different conditions.     

A morphological study on gills of the brown shrimp, Penaeus aztecus

The gills of Penaeus aztecus were examined by light and electron microscopy. They are dendrobranchiate, consisting of a central axis with biserially arranged branches that subdivide into bifurcating filaments. A septum divides the lumina of these structures into afferent and efferent channels. Hemolymph from the sternal sinus flows through the afferent channels into the filaments where it is directed into the efferent channels and finally to the pericardial cavity. In addition to these channels, numerous blood vessels permeate the gill. The cuticle covering the gill overlies a thin epithelium which is separated from hemolymph by a basal lamina. The epithelium, which is active in cuticle secretion, has a series of pillar processes that form subcuticular lacunae. The apical membranes of epithelial cells become folded in shrimp exposed to hypo- and hyperosmotic salinities. Granular cells that contain elaborate Golgi apparati and several types of granules are present throughout the gill. Nephrocytes resembling glomerular podocytes line the efferent channels. A large nerve traverses the septum in the axis.     

盐度对大麻哈鱼幼鱼存活率、鳃ATP酶活力及其组织结构的影响

为研究不同盐度对大麻哈鱼幼鱼存活率、鳃ATP酶活力及其组织结构的影响, 试验共设置4个盐度组(S0、S8、S16、S24), 试验周期42d, 解剖取鱼鳃测定ATP酶活力, 并运用组织切片及扫描电镜技术观察其鳃组织结构的变化。结果显示: S8和S16组大麻哈鱼幼鱼存活率最高, 均达到98.89%, S0组存活率为94.45%, 而S24组存活率最低, 为83.34%。随着盐度的升高, 大麻哈鱼幼鱼鳃组织Na+/K+-ATP和Ca2+/Mg2+-ATP酶活力均呈现出先升高后下降的趋势且酶活力最高的均为S8组、最低的为S24组。鳃丝宽度随盐度升高逐渐增大且各组之间呈现出显著性差异(P<0.05), 而鳃小片长度和宽度均随盐度升高逐渐减小。扫描电镜结果表明随着盐度的升高线粒体丰富细胞数量逐渐增多、顶膜变小且微绒毛消失; 同S0组相比, S8组和S16组鳃丝表面扁平上皮细胞之间的轮廓更加清楚且环形微脊条纹清晰, 而S24组鳃丝表面扁平上皮细胞之间界限模糊, 环形微脊间有融合或间断情况; 鳃小片底部扁平上皮细胞表层结构由清晰逐渐变得混乱、气孔数量逐渐减少且孔径变小。因此推测大麻哈鱼幼鱼在降海阶段适宜的盐度生存范围可能介于8‰—16‰, 具体有待进一步研究。     

Primary cell culture from gill explants of rainbow trout

Primary cultures of gill cells were initiated from gill filament explants of rainbow trout, Oncorhynchus mykiss . The explants were cultured in Leibovitz l -15 medium with 5, 10 or 20% foetal calf serum (FCS) and l -glutamine. The attachment efficiency was serum-dependent though increased FCS concentration did not stimulate further outgrowth of cells. The explants produced cell outgrowth 24 h after attachment as a sheet of cells which exhibited characteristics of gill pavement epithelial cells as indicated by surface microridges revealed by scanning electron micrographs. There was high proliferation for the first 14 days then a stable plateau for 30 days followed by a decline phase from 45 days. Following removal of cells, the explants produced further cell outgrowth which was especially active at the proliferation phase (14 days). Removal of these cells caused the explants to produce a further proliferation of cells reaching confluence in 10–14 days. After the third cell removal cell outgrowth from explants showed migratory activity but did not develop to resemble gill epithelial cells. The use of gill explants to establish primary cultures of fish gill cells has advantages which include longevity of the culture and successive proliferations from explants which could provide a useful tool for the investigation of long-term processes in cellular biology and reduce the number of culture preparations.     

Scanning electron microscopy of the respiratory surfaces of Saccobranchus (=Heteropneustes) fossilis (Bloch)

Summary The gill secondary lamellae are generally covered with epithelial cells whose outer surfaces form numerous microvilli. The surface of the primary lamellae is characterised by microridges. A particular type of surface sculpturing seems to be associated with given cell boundaries.Further evidence for the derivation of the air tube and fans which guard its entrance by modification of the basic gill structure has been obtained from both the gross surface architecture and microstructure of the individual cell surfaces. Secondary lamellae are represented by stubby projections which generally have a biserial arrangement. The outer surfaces of the epithelia overlying the capillaries of these respiratory islets are coated with microvilli as in the secondary lamellae. On the other hand, the relatively smooth-surfaced lanes between groups of respiratory islets have a microridged surface similar to that of the primary gill lamellae.It is suggested that previous estimates of surface area, and consequently diffusing capacities of the air-breathing organ, have been low in view of the increased surface, due to both their gross and microstructure. Estimates for gill surface area may need very little correction as the spaces between the microvilli and microridges are probably filled with mucus under normal conditions.We thank Mr. John Clements for his excellent technical assistance and the Department of Botany, Bristol University for the use of their scanning electron microscope     

合浦珠母贝鳃的显微与超微结构

合浦珠母贝(Pinctada fucata)是典型的滤食性瓣鳃类动物,也是我国重要的海水珍珠养殖贝类。本研究用光学显微镜、扫描电镜和透射电镜观察了合浦珠母贝鳃的显微和超微结构。结果表明,合浦珠母贝鳃结构属于异丝鳃型,左右两侧各2个鳃瓣,每个鳃瓣由内鳃瓣和外鳃瓣组成。鳃瓣由主鳃丝和普通鳃丝构成,主鳃丝在鳃瓣中主要起支架作用,每2根主鳃丝之间的9～12根普通鳃丝由"簇内连接"(intrabunchial junction)相连成簇。普通鳃丝之间通过"丝间连接"(interfilament junction)相连,丝间连接的上皮细胞与普通鳃丝的扁平细胞结构一样,为鳃的呼吸上皮。丝间连接的存在扩大了鳃的表面积,这种结构有助于进行气体交换。主鳃丝和普通鳃丝表面有前纤毛和侧纤毛,与食物运送和气体交换有关。普通鳃丝表面的纤毛为典型的"9+2"型微管结构。     

斑马鱼鳃的光镜和透射电镜观察

应用光学显微镜和透射电镜对斑马鱼(Danio rerio)鳃的组织结构及鳃丝、鳃小片超微结构进行了观察。结果表明,斑马鱼有4对全鳃,鳃耙呈长锥状,鳃丝呈梳状排列在鳃弓上,鳃小片均匀排列在鳃丝两侧。鳃小片由上皮细胞、柱细胞、内皮细胞和毛细血管网组成,鳃小片基部和血管周围分布有泌氯细胞,胞内有丰富的线粒体和排泄小泡,根据线粒体形态特征和细胞质电子密度可将其分为两个亚型。黏液细胞通常与泌氯细胞对生存在,并且有通外的开口。斑马鱼鳃组织结构与其他硬骨鱼鳃结构相似,其结构和功能有密切的关系。     

Manganese tissue accumulation and tyrosine hydroxylase immunostaining response in the Neotropical freshwater crab, <Emphasis Type="Italic">Dilocarcinus pagei</Emphasis>, exposed to manganese

Manganese (Mn) is an essential metal for the development and function of the mammalian brain; however, excess Mn accumulation may cause neurological abnormalities resembling Parkinson’s disease due to reductions in brain dopamine levels. Because dopamine also regulates many functions in crustaceans, this study examined the effects of Mn accumulation in Dilocarcinus pagei, a Neotropical freshwater crab. Following a 72-h exposure to graded concentrations of MnCl₂, Mn accumulation was assessed in several tissues. Glycaemia and the tyrosine hydroxylase (TH) immunostaining response were also examined as indicators of catecholaminergic function and catecholaminergic cell integrity, respectively. Tissue Mn accumulation was variable and occurred in the following order: gills > hepatopancreas > claw muscle > haemolymph. Exposure to 2 mM Mn reduced the gill levels of calcium, copper and iron, whereas Mn at all concentrations decreased zinc levels. All Mn-exposed animals showed lower copper levels in the hepatopancreas and haemolymph. Exposure to 2.0 mM Mn increased the haemolymph calcium. Mn exposure had no effect on glycaemia, whereas exposure to low Mn concentrations reduced the TH immunostaining response. Analysis of the central nervous system revealed the greatest Mn effect in the cerebral ganglion and the least effect in the abdominal ganglia. These results suggest the operation of an adaptive mechanism for tissue accumulation that could be responsible for the lack of an association between Mn concentrations and metal accumulation. The findings also suggest that Mn, calcium, iron and zinc share a transporter in gill cells and that Mn resistance is greater in the TH-positive cells of this crustacean than in mammalian cells.     

The Posttentacular Syncytia of Temnocephalids: the First Indication of a Putative Osmoregulatory Organ in the Epidermis of a Platyhelminth

Abstract A small syncytium or a pair of syncytia is present in the dorsal epidermis of the four species studied (Diceratocephala boschmai, Temnocephala dendyi, Craspedella sp., and Achenella sathonota). Unlike the other epidermal syncytia, the posttentacular ones may be deeply stained with silver nitrate. The basal zone of the posttentacular syncytia is occupied by abundant deep invaginations of the basal cell membrane with densely packed mitochondria between them. In one species, A. sathonota, the nuclei containing portions of the posttentacular syncytia are insunk into the parenchyma. Ultrastructurally, the posttentacular syncytia are similar to the excretory bladder (the distal portion of the protonephridial canal) of the same animals and to osmoregulatory epithelial chloride cells known from many animals. These data suggest that ion transport in this syncytium serves for osmoregulation and maintenance of ion balance. Despite extensive studies of the epidermis, no hints of such cells have ever been reported in other platyhelminths. The accessory osmoregulatory organ is probably an adaptation to life in the gill chamber of the crustacean hosts which are characterized by high ability to maintain osmotic and ionic balance in stressful conditions.     

Characterization of abalone Haliotis tuberculata–Vibrio harveyi interactions in gill primary cultures

The decline of European abalone Haliotis tuberculata populations has been associated with various pathogens including bacteria of the genus Vibrio. Following the summer mortality outbreaks reported in France between 1998 and 2000, Vibrio harveyi strains were isolated from moribund abalones, allowing in vivo and in vitro studies on the interactions between abalone H. tuberculata and V. harveyi. This work reports the development of primary cell cultures from abalone gill tissue, a target tissue for bacterial colonisation, and their use for in vitro study of host cell—V. harveyi interactions. Gill cells originated from four-day-old explant primary cultures were successfully sub-cultured in multi-well plates and maintained in vitro for up to 24 days. Cytological parameters, cell morphology and viability were monitored over time using flow cytometry analysis and semi-quantitative assay (XTT). Then, gill cell cultures were used to investigate in vitro the interactions with V. harveyi. The effects of two bacterial strains were evaluated on gill cells: a pathogenic bacterial strain ORM4 which is responsible for abalone mortalities and LMG7890 which is a non-pathogenic strain. Cellular responses of gill cells exposed to increasing concentrations of bacteria were evaluated by measuring mitochondrial activity (XTT assay) and phenoloxidase activity, an enzyme which is strongly involved in immune response. The ability of gill cells to phagocyte GFP-tagged V. harveyi was evaluated by flow cytometry and gill cells-V. harveyi interactions were characterized using fluorescence microscopy and transmission electron microscopy. During phagocytosis process we evidenced that V. harveyi bacteria induced significant changes in gill cells metabolism and immune response. Together, the results showed that primary cell cultures from abalone gills are suitable for in vitro study of host-pathogen interactions, providing complementary assays to in vivo experiments.     

Localization of the bioluminescence system in the pileus of Mycena chlorophos

Mycena chlorophos, which is primarily distributed in Southeast Asia, is a luminous fungus that emits a bright green light from its pileus for about 2 days at approximately 20°C and high relative humidity. The distribution of bioluminescent tissues in the whole pileus and its sections was heterogeneous. The light intensity in the cap and the upper region of the gill was greater than that in the lower region of the gill. At the microscopic level, the light was predominantly emitted from the membranes of hymenium and basidia cells on the gill. The emission was both cell and region specific. The luminescence system was localized in the cell membrane, and a part of the system was on the cell membrane surface. Copyright © 2015 John Wiley & Sons, Ltd.     

鲻和鲮鳃丝的扫描电镜比较观察

对鲻（Mugil cephalus）和鲮（Cirrhina molitorella）的鳃丝表面结构进行了扫描电镜比较观察,结果表明,鲻鳃丝杆状部比鲮粗．鲻鳃小片高度比鲮低;两者鳃丝表面分泌孔洞口径和密度不同;鲻和鲮细胞外被不同,鲻细胞外被稀疏,鲮的则致密复杂;鳃小片细胞和鳃丝表皮细胞的表面形态存在差异,文章还描述了鳃丝表皮形态特异的细胞。     

A preliminary report on complex fibrillar aggregates in the gill epithelium of Panulirus interruptus (Randall)

Complex structures have been found in gill epithelium of the spiny lobster, Panulirus interruptus. These fibrillar structures are located at the base of gill filaments and at the ends of longitudinal septa which divide the gill filaments into channels for directional blood flow. The complex fibrillar aggregates compare favorably with secretory granules seen in crustacean integument and are believed to play a role in chitin metabolism.     

Mechanism of Coagulation in Gecarcinus lateralis

Agglutination of cells, degranulation, and loss of cellularmembranes compose the major form of coagulation in the hemolymphof Gecarcinus lateralis. It is only after agglutination of theformed elements of the hemolymph that fibrin-like strands appear.Sodium citrate, in a concentration of 10% or more to preventcoagulation, is always inadequate to prevent cell agglutination. Multiple studies by protein electrophoresis failed to revealany differences between plasma and serum, nor did they allowus to identify a soluble protein in plasma that did not appearin serum. Crab hemolymph changes in its capacity for clottingduring the molt cycle, with the most rapid clotting occurringin the premolt period. A new protein appears in the premoltperiod, but its relation to the whole clotting mechanism isunknown. In contradistinction to vertebrate systems, citrated hemolymphdoes not clot when calcium is added. There is no relationshipthat can be demonstrated between activating systems in vertebrateplasma and clotting in the crab. It would seem that, ratherthan the vertebrate coagulating system evolving from the crustaceantype of clotting system, the development of these clotting systemshas run in parallel. The crustacean cell, in addition, appearsto be more potent than vertebrate cells in clotting systems.The comparison of human lymph to crustacean hemolymph wouldindicate that, for a given amount of cells, crustacean hemolymphclots 2 to 20 times faster than human lymph. On the other hand,agglutination of cells is a fundamental initiating step in coagulationof both human blood and crustacean hemolymph.     

A mechanism for acute aluminium toxicity in fish

Aluminium is acutely toxic to fish in acid waters. The gill is the principal target organ and death is due to a combination of ionoregulatory, osmoregulatory and respiratory dysfunction. The toxic mechanism has hitherto received little direct consideration and is unknown. In this paper the mechanism of acute aluminium toxicity is approached from a chemical perspective. Symptomatic evidence of toxicity is taken from the literature and combined with our own research to elucidate a biochemically sound model to describe a possible mechanism of acute aluminium toxicity in fish. The proposed model delineates the chemical conditions immediately adjacent to the gill surface and emphasizes their importance in aluminium's toxic mode of action. The mechanism is shown to be bipartite. Aluminium binding to functional groups both apically located at the gill surface and intracellularly located within lamellar epithelial cells disrupts the barrier properties of the gill epithelium. The concomitant iono- and osmoregulatory dysfunction results in accelerated cell necrosis, sloughing and death of the fish. The mechanism of epithelial cell death is proposed as a general mechanism of aluminium-induced accelerated cell death.     

Blood and other possible inflammatory cells in the sparid Acanthopagrus australis (Günther)

The peripheral blood cells of Acanthopagrus australis include erythrocytes, elongated and rounded thrombocytes, lymphocytes, neutrophils and monocytes. The ultrastructure of lymphocytes, neutrophils and monocytes is described. Type 1 cells with characteristic elongated, eosinophilic granules and type 2 cells with rounded, apparently labile granules are common in the gill filaments and do not increase or decrease significantly in abundance in ectoparasite-infested gill filaments. Extravascular neutrophils tend to be more evident in pathologic tissue. Lymphocytes and the macrophage-like type 3 cells are the most abundant infiltrating cells in pathologic gill tissue. Apart from ultrastructural and histochemical data, granule length and width are sufficient criteria to distinguish between the neutrophil, type 1, type 2 and type 3 cells.     

Vibrio harveyi hemolysin induces ultrastructural changes and apoptosis in flounder (Paralichthys olivaceus) cells

Vibrio harveyi hemolysin (VHH) is considered a major pathogenic virulence factor to fish. However, the VHH active-site mutant has lost all hemolytic and phospholipase activities as well as pathogenicity. In this study, the effect of VHH on erythrocytes and a gill cell line from flounder was elucidated. Erythrocyte membranes formed thin tubular protrusions immediately after exposure to VHH, and membrane corrugations were evident after extended incubation. In contrast, the mutant VHH did not induce any gross morphological changes. With VHH-treated FG-9307 cells, a cell line derived from flounder gill, destruction of organelles and formation of features resembling apoptotic bodies were observed. Immunogold staining showed that a large amount of VHH was deposited on the membranes and membrane debris of erythrocytes and FG-9307 cells after treatment with VHH. Apoptotic features, such as chromatin condensation and apoptotic bodies, were observed in VHH-treated FG-9307 cells using DAPI staining. Moreover, cell cycle analysis showed that VHH increased the proportion of cells in G1 phase. In addition, VHH significantly increased the percentage of apoptosis, the number of TUNEL positive apoptotic cells, and caspase-3 activity in FG-9307 cells when compared with the untreated controls. These data suggested that VHH killed the cells through apoptosis via the caspase activation pathway.     

Surface ultrastructural changes in the gills of sockeye salmon (teleostei: Oncorhynchus nerka) during seawater transfer: Comparison of successful and unsuccessful seawater adaptation

Sockeye salmon were transferred rapidly from freshwater to seawater and the changes in gill morphology, in particular the distribution and sizes of chloride and mucous cells on the afferent filamental surface examined. Salmon that successfully adapted to seawater were compared with salmon that did not adapt to seawater and died as a consequence of osmoregulatory failure. The number of mucus cells (density), determined from scanning electron microscopy, increased significantly after seawater challenge. A greater increase in mucus cell density occurred in the salmon that failed to adapt to seawater. Light microscopy of transverse sections of gills detected no difference in mucus cell numbers after seawater challenge. It is proposed that mucus cells that lie just beneath the gill epithelium are activated in response to the seawater challenge, and migrate and open onto the epithelium. Freshwater-adapted salmon that had low densities of chloride cells prior to the seawater challenge failed to adapt, whereas salmon that had high densities of chloride cells adapted successfully to seawater. In the latter, the density of chloride cells on the afferent surface decreased after 30 days in seawater. The apical surface of the chloride cells of freshwater-adapted sockeye were either smooth or covered with microvilli. A greater proportion of microvilli-covered chloride cells occurred in the freshwater-adapted salmon that subsequently adapted to seawater.