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.     

Cryopreservation of marine microalgae and potential toxicity of cryoprotectants to the primary steps of the aquacultural food chain

Cryopreservation, a technique of high potential for culture collections, might offer a solution for reliable supply of microalgae in aquaculture units. Marine microalgae used in aquaculture were cryopreserved under 4, −20 and −80 °C using common cryoprotectants (methanol, dimethylsulfoxide (DMSO), propylene glycol and polyvinylpyrrolidone (PVP)) with promising results for Chlorella minutissima, Chlorella stigmatophora, Isochrysis galbana and Dunaliella tertiolecta. As cryoprotectants usually are toxic above certain concentrations and exposure time, and assuming that low amounts of cryoprotectants will remain in regenerated cultures, an experimental scheme was employed to explore the lower limits of safety for these algae and their primary consumers in hatchery food chains. Results showed that methanol was well tolerated by C. stigmatophora and D. tertiolecta up to a concentration of 1.6% (v/v) while I. galbana could not survive in culture at any concentration and C. minutissima exhibited some 30% of the control's yield at 0.2%. DMSO was highly tolerated up to 1.0% by all strains with the Chlorella strains surviving well up to 2%. Propylene glycol was not only tolerated up to 8% by Dunaliella but induced mixotrophic growth as well, while for Isochrysis it was lethal at any concentration. Among zooplanktonic consumers, brine shrimp Artemia nauplii could tolerate very high concentrations of the tested cryoprotectants, the rotifer Brachionus plicatilis was found sensitive to low amounts of PVP, while the nauplii of the shrimp Penaeus japonicus and the crab Eriocheir sinensis were in general very sensitive to all cryoprotectants and in several cases to much lower amounts than 1%. However, as long as the residues of cryoprotectants are kept below 1% in the regenerated cultures, there will be no problem with the animal consumers.     

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.     

Biofouling as a reservoir of Neoparamoeba pemaquidensis (Page, 1970), the causative agent of amoebic gill disease in Atlantic salmon

Amoebic gill disease (AGD) is currently the most important disease affecting the Tasmanian salmonid industry and is caused by a marine amoeba, Neoparameoba pemaquidensis (Page, 1970). In this study biofouling communities on salmon cages were surveyed for the presence of the disease agent over a period of 4 months. Malt–yeast–seawater (MYS) agar plates were used to culture N. pemaquidensis with its presence confirmed by immunofluorescent antibody test (IFAT). Positive percentages of categorised samples ranged from 0% to 55%. The survey detected the presence of N. pemaquidensis on a number of macrofouling species (in particular bryozoan Scrupocellaria bertholetti and solitary ascidian Ciona intestinalis), and in microfouling and water samples. High percentages of positive IFATs occurred in microfouling aggregates, the solitary ascidian, C. intestinalis, and centrifuged water samples. No positive IFATs occurred from samples of Caprella sp. The presence of N. pemaquidensis was sporadic and varied in species and over sampling month. Experimental exposure of Atlantic salmon, Salmo salar, to lightly fouled netting was conducted to assess the potential for microfouling to act as a source of infection. No signs of the disease were detected in fish exposed to lightly fouled netting treatments, while 100% of positive control fish were infected and had an average of 4.24±1.79 amoebae per field of view in IFAT of mucus smears. When combined with N. pemaquidensis loads in the water column, the loads of amoebae in biofouling communities may contribute to disease outbreaks. Thus, biofouling should be considered a risk factor for AGD outbreaks.     

水温对淡水温水性鱼类生命活动的影响

水温是影响鱼类生命活动的重要生态学因子。综述水温对淡水温水性鱼类的影响,为鱼类资源的增殖和水体生物多样性保护提供参考。从繁殖、摄食生长、代谢速率、临界游泳能力、免疫功能5方面的鱼类生命活动进行综述,其中,水温对鱼类的繁殖和生长影响最大,当水温适宜时,多数淡水温水性鱼类的各项生理活动都能正常进行;但当水温超过临界温度时会导致机体内代谢紊乱、活动能力降低、甚至造成鱼类死亡,最终引起鱼类种群结构和数量分布的显著变化。鱼类的临界游泳能力和高强度运动之后的重复运动能力也会受到水温的影响;鱼类免疫应答受水温影响较大,水温严重不适会导致免疫防御机制紊乱。水温只是影响鱼类生存的生态因子之一,但在航道建设、河流生态修复、鱼类资源评估以及水生态系统保护过程中均占据不容忽视的地位。在河流开发以及水文调节过程中,要最大限度降低其对水温生态因子的影响。     

Qualitative modelling of the kelp forest of Lessonia nigrescens Bory (Laminariales: Phaeophyta) in eulittoral marine ecosystems of the south–east Pacific: an approach to management plan assessment

This contribution aims at the assessment of the sustainability of a management plan for Lessonia nigrescens in Chilean coastal ecosystems, specially when the harvest pressure could be increased due to that this macroalgae is considered as a food source for the abalon (Haliotis discus hannoi) farming along the Chilean coast. Three qualitative conceptual models were developed and compared; the first and second ones assuming homogeneous and heterogeneous environmental conditions, respectively, and the third expanded one considering eco-social components under the assumption of environmental heterogeneity. For all these models, the factors described by Vásquez [Bot. Mar. 38 (1995) 251] were included in the management plan for the harvesting of brown macroalgae. Based on Levins's Loop Analysis, the system will have a just weakly unstable moving equilibrium if the harvesting activities do not negatively affect the macrophyta's reproduction and recruitment processes. Therefore, coincidence of reproduction and/or recruitment with harvesting must be avoided in a putative management plan. Regarding the qualitative predictions of the system's behaviour after negatively affecting reproduction and recruitment, only an ecological model assuming a heterogeneous environment shows realistic responses in comparison to experimental studies on the same species. This contribution is the first approach considering the dynamical behaviour of L. nigrescens and its surroundings. Nevertheless, further models, based on the same modelling theory, may integrate more variables from social and economical fields in order to improve the scope of this complex system. Further comparative studies should focus on the assessment of the significance of the structural functions of L. nigrescens and other macroalgae for the conservation of this benthic system.     

Content of total and free amino acids in zooplanktonic food of fish larvae

The content of total and free amino acids in freshwater planktonic rotifers (Brachionus sp.), copepods (Eudiaptomus zachariasi) and groups of Daphnia pulex, Ceriodaphnia sp. and copepodites (Cyclops strenuus) of different sizes was determined. The amino acid content of Artemia salina nauplii on hatching and during fasting was also determined.

Amino acid content was lowest in rotifers and highest in stage III–IV copepodites. The major free amino acids in Cyclops strenuus dry matter were 1.43% arginine, 0.22% histidine, 0.20% alanine, 0.15% glutamic acid and 0.11% lysine. Free arginine content decreased in the daphnids as they increased in size. The content of all free amino acids in fasting Artemia nauplii was lower than in the freshwater zooplankters. The major free amino acids in nauplii were 0.55% proline, 0.41% alanine, 0.34% glycine and 0.37% serine. The content of most free amino acids in nauplii decreased during fasting. The significance of the results are discussed in relation to essential amino acid requirements of fish, and nutrition of fish larvae without fully developed gastrointestinal systems.     

Zebrafish (Danio rerio) as an animal model for bath infection by Flavobacterium psychrophilum

Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease and rainbow trout syndrome in freshwater salmonid fish worldwide, generating injuries and high mortality rates. Despite several studies on this bacterium, the infection mechanism remains unknown due to limitations in the employed animal models. In this work, we propose using zebrafish (Danio rerio) as a model for studying bacterial pathogenicity. To substantiate this proposal, zebrafish infection by F. psychrophilum strain JIP 02/86 was characterized. Zebrafish larvae were infected using the bath method, and morphological changes and innate immune system activation were monitored using transgenic fish. Salmonid-like infection phenotypes were observed in 4.74% of treated larvae, as manifested by fin, muscle and caudal peduncle damage. Symptomatic and dead larvae accounted for 1.35% of all challenged larvae. Interestingly, infected larvae with no infection phenotypes showed stronger innate immune system activation than specimens with phenotypes. A failure of function assay for myeloid factor pu.1 resulted in more infected larvae (up to 43.5%), suggesting that low infection rates by F. psychrophilum would be due to the protective actions of the innate immune system against this bacterium in zebrafish larvae. Our results support the use of zebrafish as an infection model for studying F. psychrophilum. Furthermore, the percentage of infected fish can be modulated by disturbing, to varying extents, the differentiation of myeloid cells. Using this evidence as a starting point, different aspects of the infection mechanism of F. psychrophilum could be studied in vivo.     

Efficacy of bithionol as an oral treatment for amoebic gill disease in Atlantic salmon Salmo salar (L.)

This study examined the efficacy of bithionol as an oral treatment for Atlantic salmon Salmo salar affected by amoebic gill disease (AGD). The current commercial management strategy of AGD is a costly 3 h freshwater bath. It is labour intensive and the number of baths needed appears to be increasing; hence, there is an effort to identify alternative treatments. Efficacy was examined by feeding AGD-affected Atlantic salmon twice daily to satiation with bithionol, an antiprotozoal, at 25 mg kg^− 1 feed. Three seawater (35‰, 17 °C) re-circulation systems were used each consisting of three tanks containing 32 Atlantic salmon smolts with an average (± SEM) mass of 90.4 g (± 5.2). Three feeds were examined in the trial including bithionol, plain commercial control and oil coated commercial control. Feeding commenced 2 weeks prior to exposure to Neoparamoeba spp. at 300 cells L^− 1 and continued for 28 days post-exposure. Efficacy was determined by examining gross gill score and identifying percent lesioned gill filaments twice weekly for 4 weeks post-exposure. Bithionol when fed as a two-week prophylactic treatment at 25 mg kg^− 1 feed delayed the onset of AGD pathology and reduced the percent lesioned gill filaments by 53% and halved the gill score from 2 to 1 when compared with both the plain and oil controls during an experimental challenge. There were no palatability problems observed with mean feed intake of bithionol over the trial duration with fish consuming higher levels of the bithionol diet compared to both the oil and plain controls. This study demonstrated that bithionol at 25 mg kg^− 1 feed, when fed as a two-week prophylactic treatment for Neoparamoeba spp. exposure, delayed and reduced the intensity of AGD pathology and warrants further investigation as an alternative to the current freshwater bath treatment for AGD-affected Atlantic salmon.     

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.     

Acid water and aluminium exposure: experimentally induced gill lesions in brown trout, Salmo trutta L.

Abstract. Brown trout gills were examined after exposure to different aluminium concentrations (50, 200,500μg/1) at pH 5.5 and 7.0 for up to 6 weeks. The influence of humus (Pt 0, Pt 20) and water temperature (2.5°C, 15°C) was recorded. The gill structure was studied by light, scanning and transmission microscopy, together with a complementary morphometric analysis of centrally located secondary lamellae. Low water temperature (2.5°C) resulted in advanced gill lesions after exposure to aluminium at pH 5.5. The lesions were characterized by enlargement of the secondary lamellae due to an increased number of chloride cells in the epithelia. Furthermore, the chloride cells contained cytoplasmic aluminium precipitates. Addition of humus (Pt20) or increased pH (7.0) reduced or inhibited the effects of aluminium on the gills. A high water temperature (15°C) reduced the early gill lesions observed after exposure to water at 2.5°C. Prolonged exposure to a temperature of 15°C resulted in alterations in the gill structure in all groups, including the control group.     

Therapeutic effects of enrofloxacin in an experimental infection with a luminescent Vibrio harveyi in Artemia franciscana Kellog 1906

Although a wide range and number of chemotherapeutants have been developed and applied in aquaculture, there have been no studies on the absorption of antibiotics by crustacean larvae and, consequently, none has been reported to treat an established infection.

The aim of this study was to evaluate the therapeutic effects of enrofloxacin during an experimental infection with the strain PN9801 identified as Vibrio harveyi in Artemia franciscana.

Four treatments were used: (1) Nauplii experimentally infected with V. harveyi; (2) Nauplii enriched with enrofloxacin; (3) Nauplii enriched with enrofloxacin and, 4 h later, experimentally infected with V. harveyi; and (4) Nauplii experimentally infected with V. harveyi and 24 h later enriched with enrofloxacin. Survival rates were estimated after 48 h and they were as it follows: 29%, 99%, 88% and 85% for treatments 1, 2, 3 and 4, respectively.

This study is the first one to show some evidence that antibiotics can, in fact, withhold the course of a bacterial infection already established in Artemia and possibly in crustaceans.     

Efficacy and toxicity of oxidative disinfectants for the removal of gill amoebae from the gills of amoebic gill disease affected Atlantic salmon (Salmo salar L.) in freshwater

Amoebic gill disease (AGD) of Atlantic salmon is treated commercially by bathing affected fish in freshwater. Recently, the efficacy of freshwater bathing has been questioned, and the aim of this study was to examine the potential for improving bathing efficacy using additives to the freshwater bath. AGD‐affected Atlantic salmon were bathed in 350 L tanks containing oxygenated freshwater to which chlorine dioxide (0–50 mg L^?1), chloramine‐T (0–50 mg L^?1) or hydrogen peroxide (0–100 μL L^?1) was added. Before and following a 3‐h exposure to the freshwater and chemical additive, the gills were removed from a sub‐sample of fish and the number of live amoebae on the gills were counted and smears made for confirmation of the presence of Neoparamoeba pemaquidensis, the causative agent of AGD. Following a further 3‐h exposure, a sub‐sample of fish was bled from the caudal vein and the gills were removed for histological examination. Chlorine dioxide and chloramine‐T at 25–50 and 10–50 mg L^?1, respectively, reduced the number of amoebae on the gills by approximately 50% compared with pre‐exposure numbers. The results from hydrogen peroxide treatment were equivocal and the toxicity of hydrogen peroxide was high. The toxicity of chlorine dioxide varied with freshwater hardness and/or suspended solid load, whereas chloramine‐T toxicity was low, with mortalities attributable only to elevated temperatures at the highest concentration tested. In conclusion, chlorine dioxide and chloramine‐T show promise as potential freshwater additives for the improved removal of N. pemaquidensis and possibly, other amoebae from the gills of commercially farmed Atlantic salmon.     

Potential impacts of a recent and rapidly spreading coloniser of Australian freshwaters: Oriental weatherloach (Misgurnus anguillicaudatus)

Abstract –  Oriental weatherloach ( Misgurnus anguillicaudatus ), a benthic freshwater fish species native to East Asia, was first recorded in Australian freshwater systems in 1984. Although this species is widely established around the world, very little is known about its potential impacts as an invader. We gathered field data to parameterise a mesocosm experiment investigating the impacts of Oriental weatherloach in Southeastern Australia. During this experiment, Oriental weatherloach caused significant reductions in macroinvertebrate numbers and biomass. Additionally, Oriental weatherloach were associated with elevated ammonia, nitrate/nitrite (NO _x ), and turbidity levels. Because of issues involved in scaling up mesocosm experiments we are not able to conclusively determine that Oriental weatherloach are having large impacts in natural systems. Our results come from mesocosms designed to mimic natural conditions, however, and they thus indicate that ecological impacts may be occurring, and underline the need for further research and control.     

Effects of dissolved oxygen on hemolymph parameters of freshwater giant prawn, Macrobrachium rosenbergii (de Man)

Profiles of changes in physiological parameters of freshwater giant prawns, Macrobrachium rosenbergii, exposed to various dissolved oxygen (DO) levels of 7.75, 4.75, 2.75, and 1.75 mg l⁻¹ are reported. The parameters involved in osmoregulation and oxygen transport were monitored for a 6-day period. Notable depressions in hemolymph osmolality, Na⁺, K⁺, and Cl⁻ contents were observed within 24 h after exposure to hypoxia at 2.75 and 1.75 mg O₂ l⁻¹, and thereafter remained at rather steady levels, which were significantly lower than those under normoxic conditions (4.75–7.75 mg O₂ l⁻¹). The extent of depression of osmotic-related constituents, hemolymph osmolality and Cl⁻ in particular, increased with decreased DO.

Oxyhemocyanin constituted 65.46–65.84% of total hemolymph proteins under the various DO levels examined; both hemolymph oxyhemocyanin and proteins showed notable elevations 24 h after exposure to hypoxic conditions, and reached the highest and constant level by 48 h after exposure. The compensatory responses of prawns to reduced O₂ were manifested by increased O₂ uptake through augmentation of hemocyanin, which results in enhancement of oxygen binding capacity of the hemolymph. In the same period, a significant surge of the respiratory products, PCO₂ and HCO₃⁻, was also demonstrated 6 h after exposure to hypoxic conditions which resulted in hemolymph alkalosis. These processes likely resulted in an increase in water influx and consequent declines in hemolymph osmolality and ion composition. Furthermore, hyperventilation and respiratory alkalosis, indicated by increased oxyhemocyanin and pH, respectively, were found to be predominant responses of M. rosenbergii to hypoxic stress.     

Seeding nets with neutral spores of the red alga Porphyra umbilicalis (L.) Kützing for use in integrated multi-trophic aquaculture (IMTA)

Nets in traditional Porphyra mariculture are seeded with conchospores derived from the conchocelis phase, and spend a nursery period in culture tanks or calm coastal waters until they reach several centimeters in length. Some species of Porphyra can regenerate the foliose phase directly through asexual reproduction, which suggests that the time, infrastructure, and costs associated with conchocelis culture might be avoided by seeding nets with asexual spores. Here, we present work from a short-term mariculture study using nets seeded with asexual spores (neutral spores) of a native Maine species of Porphyra. Porphyra umbilicalis (L.) Kützing was selected for this proof of concept research because of its reproductive biology, abundance across seasons in Maine, and evidence of its promise as a mariculture crop. We studied the maturation, release, and germination of the neutral spores to develop an appropriate seeding protocol for nets, followed by development of a nursery raceway to provide an easily manipulated environment for the seeded nets. Neutral spores were produced throughout the year on the central Maine coast; however, there was a temporal variability in the number and survival of released neutral spores, depending upon thallus position in the intertidal zone. Small thalli were strictly vegetative, but most thalli reproduced by neutral spores; sexual reproduction was absent. Neutral spores germinated quickly at 10 and 15 °C, but germination was delayed at 5 °C. Unlike some algal zygotes and spores, neutral spores of P. umbilicalis required light to germinate; however, irradiances of 25 and 100 μmol photons m^− 2 s^− 1 were equally sufficient for germination. Rafts of seeded nets were deployed in Cobscook Bay, Maine, at two distances from salmon aquaculture pens and at a control site on a nearby, fallow aquaculture site (no salmon). There was no difference in nitrogen content of harvested thalli; however, both the density and the surface area of harvested thalli were different among the sites. The possible causes of these differences are discussed in the context of potential use of P. umbilicalis in IMTA.     

A successful microbound diet for the larval culture of freshwater prawn Macrobrachium rosenbergii

A high moisture (63–71%), semi-purified microbound diet containing alginate was compared to newly hatched live Artemia nauplii as an exclusive diet for the culture of larval freshwater prawn Macrobrachium rosenbergii from 5th stage (weighted mean) through metamorphosis to postlarva. Two separate trials, representing larvae from different hatches, were conducted. Larvae were stocked at 50/l into cone-shaped vessels that contained 2 l of 12‰ seawater and were part of a temperature-controlled (28 °C) recirculating culture system. Larvae were manually fed either the live Artemia diet or the microbound diet exclusively, several times daily. After 14 days (23 days post-hatch (dph)), growth of larvae fed the microbound diet was 90% of that achieved for larvae fed newly hatched nauplii of Artemia. Survival of larvae fed the microbound diet was 77.3% and 73.3%, and was not significantly different from that of Artemia-fed larvae. Composed of readily available ingredients, the diet contains 46.2% crude protein and 37.4% lipid, is easy to prepare, and has good water stability. The diet is an economically practical alternative to the fluctuating cost, nutrient uncertainty, and labor associated with the use of Artemia nauplii hatched from cysts. The characteristics of the diet suggest good potential for successful use in the larviculture of other fish and crustacean species, in either the existing or a modified state.     

Physiological responses during stress and subsequent recovery at different salinities in adult pejerrey Odontesthes bonariensis

Mortality and physiological responses (plasma cortisol, glucose, osmolality, ions Na⁺ and Cl⁻ and hematocrit values) were examined in adult pejerrey Odontesthes bonariensis subjected to transport or crowding and subsequent recovery for 1 week at 0, 5 and 20 ppt NaCl. Two experiments were conducted using fish from different hatcheries; the following responses were consistent for both stocks. Mortality was observed only during post-stress recovery at 0 and 20 ppt NaCl but not at 5 ppt. During recovery, blood cortisol and glucose were generally higher, whereas osmolality and blood electrolytes (Na⁺ and Cl⁻) were lower at 0 ppt than at the other salinities. A salinity of 5 ppt NaCl could not prevent the rise in cortisol and glucose levels caused by the stress of transport, but during recovery, it prevented further increases in these features and/or accelerated their return to basal levels. This salinity also helped maintain stable blood electrolyte levels. During recovery at 20 ppt NaCl, osmolality and blood ions increased, whereas plasma cortisol and glucose generally decreased. Hematocrit values were lower at 20 ppt than at the other salinities. These results suggest that freshwater is not an adequate medium for post-stress recovery and that the presence of NaCl in the water either decreases the secretion of cortisol or promotes its clearance in O. bonariensis.