pHi regulation and ultrastructural analysis in cultured gill cells from freshwater or seawater-adapted trout

Primary cultures of gill cells from freshwater and seawater-adapted trout were compared. These cultures, developed from an explant technique, exhibited a similar growth. Ultrastructural comparison between cultured and in situ cells showed that most of the cells in primary culture resembled the so called 'pavement' cells, whereas chloride cells were not observed in the cultured epithelium. Several other cells types, representing a minority of cells in primary culture, were observed (mucous cells, vesicolar cells, cells with large dense granules and cells containing lysosomes). Morphological observations of cultured pavement cells from freshwater and seawater trout gills were similar, although the density of cellular organelles in cells was less under freshwater conditions. In addition to the morphological comparison, the regulation of intracellular pH in cultured cells from freshwater and seawater gills was examined. Resting pHi was not different for freshwater or seawater gill cells. A sodium-dependent and amiloride-sensitive mechanism was found in cultured cells. Under the experimental conditions used here, this mechanism was most likely a Na⁺/H⁺ antiporter in pavement cells from freshwater and seawater-adapted trout. The comparison of pHi recovery after acidification of cells from freshwater and seawater gills showed that the activity or the number of antiporters was higher for cells from seawater trout gill.     

A survey of salmon gill poxvirus (SGPV) in wild salmonids in Norway

In 2016, the Norwegian health monitoring programme for wild salmonids conducted a real‐time PCR‐based screening for salmon gill poxvirus (SGPV) in anadromous Arctic char (Salvelinus alpinus L.), anadromous and non‐anadromous Atlantic salmon (Salmo salar L.) and trout (Salmo trutta L.). SGPV was widely distributed in wild Atlantic salmon returning from marine migration. In addition, characteristic gill lesions, including apoptosis, were detected in this species. A low amount of SGPV DNA, as indicated by high Ct‐values, was detected in anadromous trout, but only in fish cohabiting with SGPV‐positive salmon. SGPV was not detected in trout and salmon from non‐anadromous water courses, and thus seems to be primarily linked to the marine environment. This could indicate that trout are not a natural host for the virus. SGPV was not detected in Arctic char but, due to a low sample size, these results are inconclusive. The use of freshwater from anadromous water sources may constitute a risk of introducing SGPV to aquaculture facilities. Moreover, SGPV‐infected Atlantic salmon farms will hold considerable potential for virus propagation and spillback to wild populations. This interaction should therefore be further investigated.     

Reduced total hardness of fresh water enhances the efficacy of bathing as a treatment for amoebic gill disease in Atlantic salmon, Salmo salar L

The current treatment for amoebic gill disease (AGD)-affected Atlantic salmon involves bathing sea-caged fish in fresh water, often sourced from local dams, for 3-4 h. In both a small-scale laboratory and an on-farm field experiment, the effects of water hardness on the efficacy of freshwater bathing were assessed. Results showed that soft fresh water (19.3-37.4 mg L(-1) CaCO3), whether it be naturally soft city mains water or artificially softened dam water, was more efficacious at alleviating AGD in affected fish than hard fresh water (173-236.3 mg L(-1) CaCO3). Soft freshwater bathing significantly reduced viable gill amoebae numbers (from 73.9 to 40.9% of total count) and significantly alleviated gill pathology, both gross and histological. Following bathing, gross gill pathological scores of soft freshwater bathed fish lagged 2 weeks behind hard freshwater bathed fish. Significant gill lesion fragmentation, and shedding of lesion-associated hyperplastic tissue, was accompanied by a significant reduction in AGD-affected gill filaments in soft freshwater bathed fish. Furthermore, soft freshwater bathing alleviated the blood plasma electrolyte imbalance seen in control (sea water) and hard freshwater bathed fish. This study showed that the use of soft fresh water for bathing AGD-affected Atlantic salmon could be an improvement to the current method of treatment. Not only does it reduce gill amoeba numbers, but also, it is of a therapeutic advantage with the potential to reduce bathing frequency.     

Atlantic salmon, Salmo salar L., previously infected with Neoparamoeba sp. are not resistant to re-infection and have suppressed phagocyte function

Previous studies have indicated that Atlantic salmon, Salmo salar L., affected by amoebic gill disease (AGD) are resistant to re‐infection. These observations were based upon a comparison of gross gill lesion abundance between previously infected and naïve control fish. Anecdotal evidence from Atlantic salmon farms in southern Tasmania suggests that previous infection does not protect against AGD as indicated by a lack of temporal change in freshwater bathing intervals. Experiments were conducted to determine if previous infection of Atlantic salmon with Neoparamoeba sp. would provide protection against challenge and elucidate the immunological basis of any protection. Atlantic salmon were infected with Neoparamoeba sp. for 12 days then treated with a 4‐h freshwater bath. Fish were separated into two groups and maintained in either sea water or fresh water for 6 weeks. Fish were then transferred to one tank with a naïve control group and challenged with Neoparamoeba sp. Fish kept in sea water had lower mortality rates compared with first time exposed and freshwater maintained fish, however, these data are believed to be biased by ongoing mortalities during the seawater maintenance phase. Phagocyte function decreased over exposure time and freshwater maintained fish demonstrated an increased ability to mount a specific immune response. These results suggest that under the challenge conditions herein described, antigen exposure via infection does not induce protection to subsequent AGD.     

Gross signs and histopathology of branchiostegal blister disease (balloon disease): an idiopathic disease of farmed Macrobrachium rosenbergii (De Man)

The giant freshwater prawn, Macrobrachium rosenbergii, is facing increased threat due to disease as its culture becomes more widespread. A disease characterized by the swelling of the branchiostegal region and deformities of the appendages, named balloon disease by farmers, has caused considerable economic loss in the Nellore region of Andhra Pradesh, India. Clinical signs of diseased animals include a voluminous hypertrophy of some gill filaments and the inner area of the branchiostegite. By histology, hypertrophied areas at the level of the gill filaments or branchiostegite had an identical structure corresponding to a large cyst filled with a fluid containing a few free haemocytes, limited on one side by the cuticle and on the other by the subcuticular epithelial layer. Analysis of the diseased prawns did not reveal any pathogenic agent leading us to conclude that the disease is idiopathic, probably due to suboptimal water quality conditions.     

Investigation of co‐infections with pathogens associated with gill disease in Atlantic salmon during an amoebic gill disease outbreak

Gill diseases are a complex and multifactorial challenge for marine farmed Atlantic salmon. Co‐infections with putative pathogens are common on farms; however, there is a lack of knowledge in relation to the potential effect co‐infections may have on pathology. The objective of this study was to determine the prevalence and potential effects of Neoparamoeba perurans, Desmozoon lepeophtherii, Candidatus Branchiomonas cysticola, Tenacibaculum maritimum and salmon gill poxvirus (SGPV) during a longitudinal study on a marine Atlantic salmon farm. Real‐time PCR was used to determine the presence and sequential infection patterns of these pathogens on gill samples collected from stocking until harvest. A number of multilevel models were used to determine the effect of these putative pathogens on gill health (measured as gill histopathology score), while adjusting for the effect of water temperature and time since the last freshwater treatment. Results indicate that between 12 and 16 weeks post‐seawater transfer (wpst), colonization of the gills by all pathogens had commenced and by week 16 of marine production each of the pathogens had been detected. D. lepeophtherii and Candidatus B. cysticola were by far the most prevalent of the potential pathogens detected during this study. Detections of T. maritimum were found to be significantly correlated with temperature showing distinct seasonality. Salmon gill poxvirus was found to be highly sporadic and detected in the first sampling point, suggesting a carryover from the freshwater stage of production. Finally, the model results indicated no clear effect between any of the pathogens. Additionally, the models showed that the only variable which had a consistent effect on the histology score was N. perurans.     

Clinical assessment of chloramine-T and freshwater as treatments for the control of gill amoebae in Atlantic salmon, Salmo salar L.

Infections of gill amoebae that manifest as amoebic gill disease (AGD) occur in Atlantic salmon in Tasmania. The treatment of choice is freshwater bathing; however, the effectiveness of this treatment has declined over time. In this experiment, cage trials of chloramine‐T (Cl‐T) to treat AGD in Atlantic salmon were conducted over 3 months, and involved an initial bath in either freshwater or seawater with Cl‐T, followed by a second bath 6 weeks later. Amoeba densities were reduced to 50–80% of original values for both treatments. Neoparamoeba sp. density was not affected by bathing, and was not significantly different over the course of the experiment. Lesion prevalence was higher for Cl‐T‐treated fish than for freshwater‐treated fish, with overall prevalence levels of 14.30±1.00% and 8.03±0.57% respectively. This was also seen for gross gill scores. In the fortnight after each of the two baths, Cl‐T‐treated fish had significantly higher lesion levels, although this difference was then resolved by 4 weeks post bathing. The use of Cl‐T in seawater is at least as effective as freshwater at reducing amoebae density, and may be a more practical alternative when freshwater is in short supply.     

Tolerance of lumpfish,Cyclopterus lumpus,to freshwater bath treatment for amoebic gill disease,Neoparamoeba perurans,infection and efficacy of different treatment regimens

The tolerance of lumpfish to freshwater treatment and efficacy over a range of salinities and treatment durations is examined. No structural and cellular changes were noted in organs, including gills and gut, sampled 48 hr posttreatment because of freshwater baths for periods of up to 5 hr exposure at 0 ppt or continuous treatment in 15 ppt water for 10 days. Behavioral indicators such as activity and ventilation frequency were also not significantly different between treatments, with the exception of the first trial where lumpfish were inactive during treatment in 0 ppt water compared with active fish in 32 ppt seawater. Amoeba numbers declined from an initial 21.5/gill arch to 0.33 after 3 hr in freshwater and to zero after 5 hr, and to five amoebas/gill arch after 3 hr in brackish water treatment (15 ppt). When fish were treated continuously for 15 days in 15 ppt water, amoeba numbers declined from 231 to 3.7/gill arch after 3 days and a mean of 1.75 amoebae after 10 days. In the last trial, no amoebas were present per gill arch after 3 hr treatment in 0 and 3 ppt bath.     

Salmon gill poxvirus,a recently characterized infectious agent of multifactorial gill disease in freshwater‐ and seawater‐reared Atlantic salmon

Gill diseases cause considerable losses in Norwegian salmon farming. In 2015, we characterized salmon gill poxvirus (SGPV) and associated gill disease. Using newly developed diagnostic tools, we show here that SGPV infection is more widely distributed than previously assumed. We present seven cases of complex gill disease in Atlantic salmon farmed in seawater and freshwater from different parts of Norway. Apoptosis, the hallmark of acute SGPV infection, was not easily observed in these cases, and qPCR analysis was critical for identification of the presence of SGPV. Several other agents including Costia‐like parasites, gill amoebas, Saprolegnia spp. and bacteria were observed. The studied populations experienced significant mortalities, which increased to extreme levels when severe SGPV infections coincided with smoltification. SGPV infection appears to affect the smoltification process directly by affecting the gills and chloride cells in particular. SGPV may be considered a primary pathogen as it was often found prior to identification of complex gill disease. It is hypothesized that SGPV‐induced gill damage may impair innate immunity and allow invasion of secondary invaders. The distinct possibility that SGPV has been widely overlooked as a primary pathogen calls for extended use of SGPV qPCR in Atlantic salmon gill health management.     

Evaluation of sodium percarbonate as a bath treatment for amoebic gill disease in Atlantic salmon

Amoebic gill disease (AGD), caused by Neoparamoeba perurans, is a major health challenge for Atlantic salmon aquaculture globally. While freshwater bathing for 2 hr is effective in reducing infection severity, there is need for more rapid and lower cost alternatives. To this end, a combination of sodium percarbonate (SPC) in freshwater was examined for its treatment efficacy. Initial in vitro studies showed a reduction in amoeba viability when exposed for 30 min to freshwater containing >500 mg/L SPC. Subsequently, AGD‐affected salmon were bathed for 30 min in 16°C freshwater containing 100, 500 or 1,000 mg/L SPC, or for 2 hr in 16°C freshwater to mimic industry practice. Treatment at the highest SPC concentration caused extensive gill damage and substantial mortality. Neither occurred to a significant extent at lower SPC concentrations. Gill pathology of surviving fish 10 days post‐treatment (dpt) was comparable to or more severe than pre‐treatment, and significantly (p < .001) more severe than in 2 hr freshwater bathed fish. N. perurans DNA was confirmed by qPCR in all treatment groups at 10 dpt. The data indicate that a 30‐min exposure to SPC in freshwater is not a suitable alternative to existing freshwater treatment of AGD.     

A mycosis of juvenile spiny rock lobster, Jasus edwardsii (Hutton, 1875) caused by Haliphthoros sp., and possible methods of chemical control

Mortalities of puerulus and juvenile rock lobsters, Jasus edwardsii (Hutton, 1875), held in shore-based ongrowing facilities at water temperatures between 10 and 18 °C were associated with infection by an invasive oomycete identified as Haliphthoros sp. Gross signs of disease included loss of appetite, lethargy and 1–3 brown/black focal necrotic lesions in the gills near insertion of the walking legs. Hyphae were observed in wet preparations of gill filaments excised from lesions. Histology of gill lesions showed hyphae inside the gill cuticle, invasion and lysis of the skeletal muscle and massive haemocyte infiltration and melanization at the base of walking legs adjacent to infected gill filaments. Lobsters over approximately 30 mm carapace length appeared resistant to infection. Death of affected lobsters usually occurred prior to or during the moult and in some cases may have been associated with secondary bacterial infection. Haliphthoros sp. was isolated in pure culture and marine agar 2216 was the medium that produced best growth in vitro . Two isolates were exposed in vitro to five chemicals to determine if an effective treatment could be found. Chemicals that interrupted the life cycle by killing zoospores or preventing sporulation included malachite green, trifuralin, formalin and copper sulphate. The appearance of the disease was associated with poor hygiene and its elimination may be achievable by improving husbandry practices.     

A review on the diseases of freshwater prawns with special focus on white tail disease of Macrobrachium rosenbergii

Freshwater prawns, particularly, the Giant Freshwater prawn Macrobrachium rosenbergii, are one of the most economically important farmed species in the world. Faced with increasing disease problems in penaeid shrimp culture, farmers turned to freshwater prawn farming. Freshwater prawns were considered relatively less susceptible to diseases. However, with intensification of culture and increased world trade of the farmed species, emerging diseases are beginning to constitute an increasingly serious health problem in freshwater prawn culture. This article is a review of the important diseases reported in freshwater prawns, with particular emphasis on the white tail disease of M. rosenbergii as it is the most important disease recorded to date and also the most well studied among the diseases of freshwater prawns. Steps to be taken for proper health management in the farming of this species is also touched upon.     

Efficacy of manual removal and ivermectin gavage for control of Salmincola californiensis (Wilson) infestation of chinook salmon, Oncorhynchus tshawytscha (Walbaum), captive broodstocks

Captive broodstocks of spring chinook salmon, Oncorhynchus tshawytscha , were initiated from collections of naturally produced parr from the Lemhi River, a tributary of the Salmon River, ID, USA. These fish were subsequently demonstrated to be infested with the copepod parasite Salmincola californiensis . The initial prevalence of visible adult parasites for 4 years of observations made shortly after collection varied from 19.7 to 71.6%. Both the prevalence and intensity of the infestation increased in the freshwater culture of these fish. Manual removal was initiated as a means of control and practiced at monthly intervals. The number of Salmincola removed decreased in the ensuing 5 months, but the prevalence was not greatly affected. Ivermectin (22,23 dihydroavermectin), was diluted with saline and delivered by gavage at the rate of 0.20 mg kg⁻¹ body weight when the groups were being handled for the manual removal of parasites. Either two or three ivermectin treatments were given to four broodstocks of chinook salmon depending on the severity of the infestation and on the extent of gill pathology. The combination of manual removal and ivermectin gavage eliminated live Salmincola and resolved all associated necrosis of the gill tissues. There was no trend to indicate that individual chinook salmon possessed a natural resistance to reinfestation.     

Further development of bithionol therapy as a treatment for amoebic gill disease in Atlantic salmon, Salmo salar L.

This study examined the efficacy of bithionol as a prophylactic or therapeutic oral treatment for Atlantic salmon (AS), Salmo salar , affected by amoebic gill disease (AGD). Furthermore, it explored the interaction of bithionol oral therapy with the current standard treatment (a freshwater bath for at least 3 h). The efficacy of three medicated feeds was determined in the trial by feeding AGD-affected AS at 1% body weight (BW) day⁻¹ either oil coated commercial feed (control) or prophylactic and therapeutic bithionol at 25 mg kg⁻¹ feed. Feeding commenced 2 weeks prior to exposure to Neoparamoeba spp. at 300 cells L⁻¹ and continued for 49 days post-exposure (PE). Bithionol when fed as a 2-week prophylactic or therapeutic treatment at 25 mg kg⁻¹ feed delayed the onset of AGD pathology and reduced the percentage of gill filaments with lesions. Administration of a 3-h freshwater bath at 28 days PE significantly reduced amoeba numbers to a similar level across all treatments; in contrast, gross gill score and percent lesioned filaments were reduced to different extents, the control having a significantly higher score than both bithionol treatments. Following the freshwater bath, clinical signs of AGD increased at a similar level across all treatments, albeit controls were significantly higher than the bithionol treatments immediately following freshwater treatment. This study demonstrated that bithionol at 25 mg kg⁻¹ feed, when fed as a 2-week prophylactic or a therapeutic treatment, delayed and reduced the intensity of AGD pathology and warrants further investigation as a treatment for AGD-affected AS.     

急性盐度胁迫对摄食不同镁水平饲料鲈血清渗透压和离子水平以及鳃丝ATP酶活力的影响

为研究饲喂不同镁水平饲料后,鲈血清渗透压、离子水平和鳃丝ATP酶活力对急性盐度胁迫的反应,实验用镁水平为0.413 g/kg(D1,对照组)、1.042 g/kg(D2)、1.577 g/kg(D3)、1.991 g/kg(D4)的4种实验饲料,分别投喂初始平均体质量为(30±0.5)g的鲈。实验鱼在淡水循环养殖系统中饲养50 d后立即转入海水循环养殖系统,并在转入前(0 h)和转入后1、3、6、12、24 h分别采集鲈鳃丝和血液样本。结果显示:盐度胁迫前饲料镁水平对鲈血清渗透压、Na+、K+、Cl-、Mg2+含量和鳃丝中Na+/K+-ATP(NKA)与Ca2+/Mg2+-ATP(CMA)酶的活力有显著影响(P<0.05),其中血清渗透压、Mg2+含量和NKA酶活力随饲料镁水平的增加而升高。D4组的血清渗透压、Mg2+含量、NKA和CMA酶活力显著高于其他组(P<0.05),表明该组鲈处于高强度的渗透压调节的生理状态。急性盐度胁迫中,血清Na+和Cl-含量随胁迫时间的延长呈逐渐上升趋势,血清渗透压、K+、Ca2+、Mg2+和鳃丝ATP酶活力在0～24 h内呈波动性变化。D1组的NKA和CMA酶活力在胁迫1 h时显著低于其他组(P<0.05),表明长期摄食低镁水平的饲料会降低鲈鳃丝NKA和CMA酶对环境盐度刺激的敏感度。研究表明,淡水养殖鲈的饲料中镁水平应低于1.991 g/kg,以减少维持鳃丝NKA与CMA酶高活力而导致的能量损失,而在环境由低盐向高盐的快速转变过程中,较高的饲料镁水平可通过快速提高的鳃丝NKA与CMA酶活力而有利于鲈快速适应高盐环境。     

Waste treatment in recirculating aquaculture systems

Recirculating aquaculture systems (RAS) are operated as outdoor or indoor systems. Due to the intensive mode of fish production in many of these systems, waste treatment within the recirculating loop as well as in the effluents of these systems is of primary concern. In outdoor RAS, such treatment is often achieved within the recirculating loop. In these systems, extractive organisms, such as phototrophic organisms and detritivores, are cultured in relatively large treatment compartments whereby a considerable part of the waste produced by the primary organisms is converted in biomass. In indoor systems, capture of solid waste and conversion of ammonia to nitrate by nitrification are usually the main treatment steps within the recirculating loop. Waste reduction (as opposed to capture and conversion) is accomplished in some freshwater and marine indoor RAS by incorporation of denitrification and sludge digestion. In many RAS, whether operated as indoor or outdoor systems, effluent is treated before final discharge. Such effluent treatment may comprise devices for sludge thickening, sludge digestion as well as those for inorganic phosphate and nitrogen removal. Whereas waste disposed from freshwater RAS may be treated in regional waste treatment facilities or may be used for agricultural purposes in the form of fertilizer or compost, treatment options for waste disposed from marine RAS are more limited. In the present review, estimations of waste production as well as methods for waste reduction in the recirculating loop and effluents of freshwater and marine RAS are presented. Emphasis is placed on those processes leading to waste reduction rather than those used for waste capture and conversion.     

In vitro survival and the effect of water chemistry and oxidative chemical treatments on isolated gill amoebae from AGD-affected Atlantic salmon

Amoebic gill disease (AGD) of cultured salmonids in Tasmania is caused by the amphizoic parasitic amoeba Neoparamoeba pemaquidensis. The freshwater tolerance of amoebae isolated from the gills of AGD-affected salmon (predominantly N. pemaquidensis) was tested in vitro using a trypan blue exclusion assay. Amoebae exposed to water containing high concentrations of Ca²⁺ or Mg²⁺ (200 mg l⁻¹) showed high levels of survival up to 3 h of exposure. Exposure to water containing elevated Na⁺, choline chloride or water at different pH all had no significant survival of amoebae. Exposure of amoebae to different concentrations of chlorine dioxide, chloramine-T or hydrogen peroxide in artificially hard water demonstrated that chloramine-T and hydrogen peroxide were the most efficacious at killing amoebae in vitro. This work suggests that the hardness of freshwater may be an important factor for the survival of marine amoebae (predominantly N. pemaquidensis) on the gills of AGD-affected salmon and have significant implications with regard to the efficacy of freshwater bathing practices for the control of AGD on farms. Additionally, chloramine-T and hydrogen peroxide appear to be efficacious at killing marine gill amoebae in vitro and may be useful for the control of AGD in farmed Atlantic salmon.     

Effects of dissolved organic carbon and hardness in freshwater used to treat amoebic gill disease

Amoebic gill disease (AGD) is a significant disease of Atlantic salmon farmed in South East Tasmania. The commercial treatment for the disease is a freshwater bath for up to 4 h. Previous studies have shown that the chemical composition of the freshwater, in particular total water hardness, affects the efficacy of the treatment. The aim of this study was to determine if other water chemistry parameters, such as dissolved organic carbon (DOC), interact with total water hardness to affect treatment success. Firstly, the relative survival of isolated gill amoebae incubated for up to 3 h with hard or soft water (346.0 and 34.6 mg L^?1 CaCO₃ respectively) with low or high concentrations of humic or tannic acid (5 and 50 mg L^?1 respectively) was determined. Secondly, fish with AGD were bathed for 2.5 h in hard or soft water (249.3 and 35.3 mg L^?1 CaCO₃) containing either 5 or 20 mg L^?1 humic acid. The number of viable amoebae surviving on the gills and number of gill lesions were determined. It was found that the concentration of DOC used in this study that represents the levels commonly found around SE Tasmania is unlikely to have any commercial significance in the reduction in amoebae on the gills of Atlantic salmon. However, this study provided further support that freshwater selected for bathing AGD‐affected salmonids should be chosen primarily on its total water hardness.     

Development of a novel histopathological gill scoring protocol for assessment of gill health during a longitudinal study in marine-farmed Atlantic salmon (Salmo salar)

Gill health and disease in farmed Atlantic salmon (Salmo salar) were studied at a marine site on the west coast of Ireland. Fish were sampled on a regular basis during the summer and autumn of 2009. A novel histopathological scoring system was devised to give a semi-quantitative measure of gill pathology. The index criteria for gill histopathology were lamellar hyperplasia, lamellar fusion, circular anomalies (necrosis or sloughing) and lamellar oedema, a score from 0 to 3 being assigned for each parameter. Ancillary criteria, such as hypertrophy, haemorrhage and the presence of specific pathogens, were assigned a score of 0 or 1. Environmental monitoring undertaken included zooplankton and phytoplankton sampling, as well as temperature and oxygen recordings for the site. Epitheliocystis was frequently observed at low levels but was not associated with any significant gill pathology. Clinically significant gill pathology was observed on one occasion during the sampling period, coinciding with the occurrence of Pelagia noctiluca, Muggiaea atlantica and Solmaris corona. Throughout the longitudinal study, the scoring system provided an accurate and effective means of assessing gill health, demonstrating the ability of this practical tool to be used in future studies of gill health and disease.     

Morphological changes in gill mitochondria-rich cells in cultured Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> acclimated to a wide range of environmental salinity

Morphological changes in gill mitochondria-rich (MR) cells were examined in cultured Japanese eel acclimated to deionized freshwater (DFW), freshwater (FW), 30%-diluted seawater (DSW), and seawater (SW). The gill Na⁺/K⁺-ATPase activity was higher in SW-acclimated eel than in those acclimated to DFW, FW, and DSW. Immunocytochemical observations revealed that MR cells in the gill filaments were most developed in SW, whereas MR cells in the lamellae were preferentially observed in DFW, suggesting that filament and lamellar MR cells are responsible for ion secretion and absorption, respectively. In scanning electron microscopic observations, the apical membrane of lamellar MR cells appeared as a flat or slightly projecting disk with a mesh-like structure on its surface. In contrast, the apical membrane of filament MR cells showed a slightly concave surface. Whole-mount immunocytochemistry revealed that most MR cells showed cystic fibrosis transmembrane conductance regulator immunoreaction in their apical region in fish in DSW and SW, but not in those in DFW and FW, indicating that MR cells developed in DSW and SW function as an ion-secreting site. In addition to MR cells, distinct Na⁺/K⁺-ATPase immunoreaction was observed in the outermost layer of gill epithelia, suggesting that pavement cells are an additional site of ion uptake in the gills.