Stress response of juvenile rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)to chemical cues released from stressed conspecifics

The objective of this study was to determine whether exposure of rainbow trout (Oncorhynchus mykiss) to water containing a stressed trout or skin extract from stressed and non-stressed trout would elicit a stress response in conspecifics. Juvenile rainbow trout were exposed for 1 hour to water containing a stressed fish, homogenized skin extracts from a non-stressed fish, skin extract from a stressed fish and water with none of these factors. The stress response was measured over a 24-h period (1, 6, 12, 24 h after exposure). Plasma cortisol levels increased at 12 h in fish exposed to water from a stressed fish and skin extract from a stressed fish. Plasma glucose and hepatic hsp70 levels were not affected by treatments. The results suggest that rainbow trout elicit a stress response when exposed to stress-related alarm cues released from conspecifics.     

亚硝酸盐胁迫对草鱼肝脏泛素-蛋白酶体系统的影响

为了探究亚硝酸盐胁迫对草鱼(Ctenopharyngodonidellus)肝脏泛素-蛋白酶体系统(ubiquitinproteasomesystem,UPS)的影响,将草鱼随机暴露于0、0.5、20mg/L的亚硝酸盐溶液中,在12、24、48、96h后采集肝脏样品,检测血液的生化指标,nrf2、hsp70和UPS相关基因的表达,泛素化蛋白的含量以及肝脏的组织学变化。结果显示:在亚硝酸盐胁迫12h,血清皮质醇含量呈现显著的剂量依赖式升高,且在24h和48h显著高于对照组。谷丙转氨酶和谷草转氨酶的含量在96h显著性降低。暴露12h时,20mg/L暴露组的nrf2、hsp70的表达量显著性高于对照组,ub、chip、psma2、psmc的表达水平和泛素化蛋白的含量在20mg/L亚硝酸盐胁迫24h均出现显著升高。在高浓度亚硝酸盐胁迫96h后,肝细胞出现明显的空泡化。研究表明,亚硝酸盐胁迫使草鱼产生了显著的应激反应,造成了肝组织损伤,使肝脏UPS活性上升。     

Effects of acidic water,aluminum, and manganese on testicular steroidogenesis in <Emphasis Type="Italic">Astyanax altiparanae</Emphasis>

Metals can influence the gonadal steroidogenesis and endocrine systems of fish, thereby affecting their reproduction. The effects of aluminum and manganese in acidic water were investigated on steroidogenesis in sexually mature male Astyanax altiparanae. Whether mature male fish recover from the effects of metals in metal-free water was also assessed. The fish were exposed to 0.5 mg L^?1 of isolated or combined aluminum and manganese in acidic pH (5.5) to keep the metals bioavailable. The fish underwent 96 h of acute exposure, and samples were taken 24 and 96 h after the beginning of the experiment. The fish were then maintained in metal-free water for 96 h. Plasma levels of testosterone, 11-ketotestosterone, 17β-estradiol, and cortisol were measured. Acidic water increased the plasma concentration of testosterone and 11-ketotestosterone. Aluminum increased the testosterone levels after 96 h of exposure. Manganese increased the 17β-estradiol levels after 24 h of exposure and maintained at high levels until the end of the experiment. With the exception of acidic pH, which increased cortisol levels after 24 h of exposure, no changes were observed in this corticosteroid during the acute experiment. Aluminum and manganese together also altered steroid levels but without a standard variation. The fish recovered from the effects of most exposure conditions after 96 h in metal-free water. A. altiparanae could use reproductive tactics to trigger changes in testicular steroidogenesis by accelerating spermatogenesis and spermiogenesis, which may interfere with their reproductive dynamics.     

Stress responses of Atlantic salmon (Salmo salar L.) elicited by water level reduction in rearing tanks

The Atlantic salmon (Salmo salar) stress response was examined by measuring plasma cortisol, glucose and chloride in fish after water level reduction within rearing tanks. Maximum plasma cortisol levels (366±43 (SD) nM and 534±280 nM for Groups 1 and 2, respectively) were observed 20 min after application of the stressor. Cortisol levels were down to control levels 24 h later. The pattern for cortisol changes observed within these two groups were comparable. Plasma glucose and chloride concentrations did not change significantly in either of the experimental groups. The fish in group 2 were then repeatedly subjected to the same stressor every third day. After the fifth exposure to the stressor, blood was obtained from fish at times corresponding to sampling after the first exposure. Their maximum plasma cortisol level only reached 223 (±96) nM, and was down to prestress levels within 2 h. Plasma glucose and chloride concentrations did not change significantly in this second experiment. This challenge test revealed an acute primary response in Atlantic salmon without any apparent harmful secondary responses, that may thus serve as a standardized reference stressor using other fish groups under comparable conditions.     

Cortisol and liver metabolism of immature American eels,Anguilla rostrata (LeSueur)

Plasma and liver composition, liver enzyme activities, and metabolite flux in isolated hepatocytes have been studied in immature American eels,Anguilla rostrata, injected daily IP with saline or cortisol (0.35 mg/kg eel). Plasma cortisol values were significantly increased above saline controls in those eels receiving cortisol at 3h and 6h following the final (tenth) injection. On day 6 and 10 of injection plasma cortisol levels were significantly below saline controls 24h following cortisol injection. Plasma glucose values were significantly depressed in the cortisol-injected eels at both 6 and 24h following the final (tenth) injection. At the 24h sampling time, plasma protein had significantly increased, but there was no change in either plasma amino acid or fatty acid levels. An increased hepatosomatic index was attributed to a major increase in total lipids, as both protein and glycogen contents were decreased. Of the liver enzymes assayed, significant activity changes occurred only for lactate dehydrogenase (decreased), mitochondrial citrate synthase (increased) and phosphoenolpyruvate carboxykinase (increased) 24h following the final (tenth) cortisol injection. Although enzyme activity changes implied increased liver gluconeogenesis, the absolute rate of lactate, alanine, and aspartate incorporation into glucose declined in viable hepatocytes isolated from cortisol-injected eels compared to the saline controls. Relative changes in metabolite flux did support a preferential increase in gluconeogenesis from amino acids. These results are consistent with the increase in amino acid gluconeogenesis as a result of cortisol administration implied in previous studies, but failed to show a definitive cortisol effect on this pathway in the eel liver. It is suggested that other hormones (e.g. thyroxine, catecholamines, glucagon) may interact in a complex way with cortisolin vivo to bring about the biochemical changes observed in this study. The rapid clearance of exogenously injected cortisol noted in this study makes causal relationships between the injected hormone and any observed metabolic effect in the intact animal difficult.     

Efficacy of Benzocaine as an Anesthetic in Juvenile Tambaqui Colossoma macropomum

Abstract.— The present study investigated the use of benzocaine as an anesthetic for juvenile Colossoma macropomum (tambaqui). In the first experiment, fish were exposed to various doses of benzocaine for 10 min at 24 C. The second experiment examined the effects of duration of exposure to 100 mg/L of benzocaine. In the third experiment, fish were exposed to 100 mg/L at temperatures of 24, 27, and 30 C. Benzocaine concentrations of 100–150 mg/L were considered ideal for quickly inducing total immobilization and fast recovery. Fish exposed to 350 mg/L benzocaine exhibited 30% mortality. No changes in hemat-ocrit were recorded in fish exposed to different concentrations of benzocaine. Plasma glucose increased significantly when fish were exposed to benzocaine concentrations greater than 200 mg/L. Recovery time after a 30-min exposure to 100 mg/L benzocaine was significantly greater than after an exposure for 10 and 20 min. No mortality was observed 96 h after exposure to 100 mg/L benzocaine for 10, 20, and 30 min. Dosages in the 100–150 mg/L range were effective for periods of up to 20 min of anesthesia. There was no effect of temperature on the time required for fish to lose equilibrium. However, recovery was significantly faster for fish anesthetized at 30 C. Benzocaine is an effective anesthetic agent for tambaqui juveniles, providing rapid immobilization and rapid recovery. Benzocaine is also less expensive than other available anesthetic compounds.     

Dietary choline requirement of juvenile olive flounder (Paralichthys olivaceus)

An eight‐week study was conducted to determine the optimum dietary choline level in juvenile olive flounder, Paralichthys olivaceus. Seven diets were prepared to contain 0, 250, 500, 750, 1,000, 2,000 and 3,000 mg/kg diet. Juveniles (5.9 ± 0.03 g; 5.5 ± 0.4 cm; mean ± SD) were randomly distributed into 21 tanks (25 fish/tank) and fed one of the diets in triplicates. Survival rate of fish fed the diet containing the lowest choline level was significantly lower than those of fish fed the other diets (p < 0.05). Final body weight, weight gain, specific growth rate, feed efficiency and protein efficiency ratio significantly increased with increasing choline levels up to 1,000 mg/kg diet. Whole‐body protein and lipid contents increased in accordance with choline levels up to 750 mg/kg diet, beyond which they plateaued. Liver and muscle lipid contents elevated with increasing choline levels up to 2,000 mg/kg diet. Plasma cholesterol esters, triglycerides, cholesterol and total lipids were significantly influenced by the graded choline levels; however, responses of those indices were not identical. Broken‐line analyses of weight gain and liver choline concentrations responding to the graded choline levels revealed that choline requirements of the juvenile flounder could be between 847 and 1,047 mg/kg diet.     

Acute Toxicity of Nitrite to Red Drum Sciaenops ocellatus: Effect of Salinity1

Acute toxicity of nitrite to red drum fingerlings was investigated under static conditions in environments containing 36.0 to 0.6 g/L salinity. The 48 h median lethal concentrations ranged from 85.7 mg/L nitrite-N (36.0 g/L salinity) to 2.8 mg/L nitrite-N (0.6 g/L salinity). Plasma nitrite concentrations increased with exposure time during a 48 h study and exceeded environmental concentrations in fish exposed to 9.1 and 5.1 mg/L nitrite-N (1.4 g/L salinity). During 24 h of exposure, methemoglobin levels increased with increasing environmental nitrite conditions in fish exposed to 3, 6 and 9 mg/L nitrite-N for 24 h (1.4 g/L salinity). The chloride component of salinity was not as effective in preventing nitrite toxicity as in other species of fish, indicating a potential problem for the culture of red drum in low-salinity waters.     

Acute toxicity of the pesticide methomyl on the topmouth gudgeon (Pseudorasbora parva): mortality and effects on four biomarkers

In this study, the acute toxicity of the pesticide methomyl on the topmouth gudgeon (Pseudorasbora parva) was evaluated using mortality and the activity of the enzymes acetylcholinesterase (AChE), glutathione S-transferases (GSTs), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) as endpoints. LC50 values were 1.228, 0.782, 0.538, and 0.425 mg/l at 24, 48, 72, and 96 h of exposure, respectively. Methomyl caused a sharp decrease in specific activity of brain AChE around 48% at concentrations between 0.043 and 0.213 mg/l. A reduction higher than 40% in liver GST activity at concentrations between 0.085 and 0.213 mg/l was found, whereas no significant effects were observed in intestinal GST. A significant concentration-dependent decrease of GOT activity was found after 24 h of exposure to the pesticide but not after 96 h. No significant effects on GPT activity were observed. These results indicate that at the concentrations tested, methomyl is acutely toxic to the species P. parva, causing mortality, neurotoxic effects, and changes in some hepatic enzymes.     

Early biochemical biomarkers for zinc in silver catfish (<Emphasis Type="Italic">Rhamdia quelen</Emphasis>) after acute exposure

Contamination of aquatic ecosystems by metals causes various biochemical changes in aquatic organisms, and fish are recognized as indicators of environmental quality. Silver catfish were exposed to six concentrations of zinc (Zn): 1.0, 2.5, 5.0, 7.5, 10.0 and 12.5 mg/L for 96 h to determine the mean lethal concentration (LC₅₀). The value obtained was 8.07 mg/L. In a second experiment, fish were exposed to concentrations of 1.0 or 5.0 mg/L Zn and a control for 96 h. Afterward, the tissues were collected for biochemical analysis. Lipid peroxidation, as indicated by thiobarbituric acid-reactive substance (TBARS), decreased in the liver and brain for all Zn concentrations tested, while in the gills TBARS levels increased at 1.0 mg/L and declined at 5.0 mg/L. Zn increased protein carbonyls in the muscle of silver catfish and decreased it in the other tissues. The enzyme superoxide dismutase increased in both exposed groups. However, catalase did not change. Glutathione S-transferase decreased in the liver and increased in the gills (1.0 mg/L), muscle (5.0 mg/L) and brain (1.0 and 5.0 mg/L). Nonprotein thiols changed only in brain and muscle tissue. Zn exposure inhibited acetylcholinesterase (AChE) activity in the brain at both concentrations tested, but did not change it in muscle. Exposure to Zn inhibited the activity of Na⁺/K⁺-ATPase in the gills and intestine at both concentrations tested. Our results demonstrate that Zn alters biochemical parameters in silver catfish and that some parameters such as AChE and Na⁺/K⁺-ATPase could be considered as early biomarkers of waterborne Zn toxicity.     

Physiological Responses of Rainbow Trout Oncorhynchus mykiss to Crowding and Anesthesia with AQUI-S™

Abstract.— Following exposure to the anesthetic AQUI-S™, plasma cortisol concentration in immature rainbow trout was measured as (mean) 293 ± 48 ng/ mL, which was significantly ( P > 0.05) higher than the mean concentration in resting fish. Cortisol concentrations remained significantly ( P > 0.05) elevated for at least 24 h after treatment. This was accompanied by a significant increase and decrease in hematocrit and plasma potassium, respectively. These perturbations continued for at least 48 h following recovery from anesthesia. Plasma concentrations of total protein and sodium remained unchanged following anesthesia with AQUI-S™. Crowding stress is commonly encountered by fish during manipulation in aquaculture situations. Anesthetising fish prior to, and during, manipulation may reduce the associated stress. Changes in cortisol values resulting from crowding (30 min; 0.1 kg/L) during anesthesia with AQUI-S™ were not appreciably different from those in fish crowded without anesthesia. Thus, anesthesia with AQUI-S™ at the recommended dose of 17 mg/L did not appear to be effective for alleviating the stress of crowding under the conditions of our experiments.     

A comparative study on the effects of a pesticide (cypermethrin) and two metals (copper, lead) to serum biochemistry of Nile tilapia, Oreochromis niloticus

The present study was designed to compare the responses in freshwater fish Oreochromis niloticus exposed to a synthetic pyrethroid, cypermethrin (CYP); an essential metal, copper (Cu); and a nonessential metal, lead (Pb). Fish were exposed to 0.05 μg/l CYP, 0.05 mg/l Cu, and 0.05 mg/l Pb for 4 and 21 days, and the alterations in serum enzyme activities, metabolite, and ion levels were determined. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities increased in response to CYP, Cu, and Pb exposures at both exposure periods. While elevations in alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) activities and in cholesterol level were observed in pesticide-exposed fish at 4 and 21 days, they increased in Cu- and Pb-exposed fish at 21 days. Although metal-exposed fish showed increases in cortisol and glucose levels at 4 days followed by a return to control levels at the end of the exposure period, their levels elevated in pesticide-exposed fish at both exposure periods. Total protein levels decreased in Pb- and pesticide-exposed fish at 21 days. Na⁺ and Cl⁻ levels decreased in pesticide-exposed fish at both exposure periods and in Cu- and Pb-exposed fish at 21 days. The exposures of pesticide and metals caused an elevation in K⁺ level at the end of the exposure period. The present study showed that observed alterations in all serum biochemical parameters of fish-treated pesticide were higher than those in fish exposed to metals.     

Effects of varying salinities on the toxicity of potassium permanganate to larval and juvenile striped bass, Morone saxatilis (Walbaum)

Abstract. Eight-day-old striped bass, Morone saxatilus (Walbaum) larvae, and one-month-old juveniles were held in varying salinities and exposed to a range of concentrations of potassium permanganate. Lethal concentrations of potassium permanganate for 50% of the larval fish exposed for96h (96h LC₅₀) were 1middot;02, 2 middot;11, 1 middot;41 and 1 middot;73mg/1 for salinities of 0, 3, 6and 9 g/l, respectively; The 96h LC₅₀S for juveniles were 0 middot;96, 3 middot;26, 1 middot;63 and 1 middot;48 mg/l for salinities of 0, 5, 10 and 15g/l, respectively. There were significant differences in 96h LC₅₀ values at different salinities for both life stages. The 96 h LC₅₀ values derived for both life stages revealed a decrease in toxicity as salinity increased from 0 to 3–5g/l. Results indicated potassium permanganate to be toxic in fresh water to striped bass larvae and fingerlings at recommended treatment levels (3 middot;0mg/l).     

In vitro evidence that cortisol directly modulates stress-related responses in the skin epidermis of the rainbow trout (Oncorhynchus mykiss Walbaum)

Exposure of fish to stressors leads to multiple changes in the skin epithelium. We investigated the role of the stress hormone cortisol in the control of these changes by exposure of pieces of skin from the rainbow trout Oncorhynchus mykiss with an in vitro tissue culture incubation procedure. The effects of 24 h exposure to 4 cortisol concentrations (0, 50, 500 and 1000 ng/ml) were determined. Numbers of mucous, mitotic and apoptotic cells were quantitatively assessed using immunohistochemical techniques, in situ DNA nick end labelling (TUNEL), as well as conventional light and scanning electron microscopy (SEM). The cortisol receptor blocker mifepristone was used to investigate whether the effects could be attributed to the direct action of the hormone via glucocorticoid receptors. Overall, cortisol had no effect on the mucous cell population at 24 h. Incubation with the receptor blocker reduced the number of mucous cells. Cell proliferation was stimulated by the addition of 50 and 500 ng/ml cortisol, but not at 1000 ng/ml. Incubation with the receptor blocker increased proliferation in the control group (0 ng/ml) only. An increase in apoptosis occurred at 500 and 1000 ng/ml cortisol. This increase was blocked by incubation with the receptor blocker, which resulted in lower numbers of apoptotic cells in all except the 0 ng/ml controls. SEM observations corroborated the quantitative data. The results indicated that the effects of stressors on the fish epidermis mentioned above are mediated by cortisol, with the exception of mucous cell release.     

Efficiency of 2‐phenoxyethanol and Clove Oil for Reducing Handling Stress in Reared Meagre,Argyrosomus regius (Pisces: Sciaenidae)

The effect of two anesthetics, 2‐phenoxyethanol (2‐PE) and clove oil (COil) were studied in meagre, Argyrosomus regius. This study aimed to determine the adequate dose of anesthesia for different aquaculture procedures, to assess a sedative or stressor effect of low anesthetic concentrations, and to assess the effect of low anesthetic doses at high fish densities for longer periods of time on fish survival and behavior. Anesthetics were tested at different concentrations: 2‐PE at 100, 250, 400, 550, and 700 mg/L; COil at 25, 40, 55, 70, and 85 mg/L. Meagre became anesthetized within 10 min when using concentrations above 250 mg/L for 2‐PE and above 40 mg/L for COil. Deep anesthesia was achieved at 700 and 85 mg/L, for 2‐PE and COil, respectively. The fish did not react when blood was collected at these concentrations. Plasma cortisol and glucose levels were similar between sedated meagre with both 2‐PE – 100 mg/L and COil – 10 mg/L, and the control (not sedated; P > 0.05). This suggests that low concentrations of these anesthetics induce similar stress response as handling without anesthesia during routine activities. No mortality was registered when meagre was maintained at high densities with low concentrations of each anesthetic for 2 h.     

Effect of sub-lethal nitrite on selected haematological parameters in fingerling Catla catla (Hamilton)

A sub‐lethal nitrite toxicity trial was conducted using static conditions for a period of 96 h with fingerlings of Catla catla (21.4±3.6 g). Fingerlings were exposed to five concentrations of nitrite, that is, 1, 2, 4, 8 and 10.4 mg L^?1 and a nitrite‐free control to study changes in haematological parameters. Nitrite caused an increase in immature erythrocyte population (7–24%) in lower concentrations (0–4 mg L^?1) at 6 h while they were absent in higher concentrations. The total erythrocyte count was reduced at 6 h followed by an increase at 12 h with further reduction up to 96 h in all concentrations of nitrite. The 96‐h exposure resulted in 21.2–31.8% reduction in erythrocyte population in 1–10.4 mg L^?1 nitrite. The haemoglobin content decreased progressively with increasing nitrite concentrations as well as exposure periods. Total leukocyte count decreased initially at 6 h in all treatments followed by an increase after 12 h, signifying development of a protective response of the body to nitrite stress. Blood glucose decreased initially up to 24 h followed by an increase through 96 h. Serum protein level decreased continuously with increasing exposure period. The study revealed that exposure to nitrite caused changes in almost all the haematological parameters in the fingerlings depending on the concentration as well as exposure period. Nitrite being one of the important inorganic nutrients often recorded at higher levels in intensively cultured ponds, the present study highlights its adverse impact on fish and stressed the need for the management of this nutrient in culture ponds.     

不同质量浓度铅对草鱼鱼种抗氧化性能的影响

将草鱼暴露于不同水平(0、72、144、288 mg/L)的硝酸铅水溶液中,分别于12、24、48、96 h测定草鱼肝、鳃组织中谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量.试验结果表明,当鱼经不同质量浓度Pb2+处理24 h后,其肝脏的GSH-Px活性表现为诱导效应;而鳃在低质量浓度(72 mg/L)Pb2+处理也表现诱导效应,但高质量浓度Pb2+(144、288 mg/L)处理24 h后,其GSH-Px活性表现为抑制效应.与肝GSH-Px相比,草鱼肝SOD经Pb2+处理后其活性抑制更加明显,表明肝SOD对Pb2+的毒性反应比肝GSH-Px敏感.同时,草鱼经不同质量浓度Pb2+处理12 h后,肝与鳃的MDA含量均比对照组明显增加.上述研究结果表明,水体铅可能通过诱导草鱼肝脏与鳃发生氧化损伤而发挥毒性作用,并进一步损伤机体结构及其生理功能.     

Acute toxicity of Trichlorfon (Dipterex) to fry of Cichlasoma urophthalmus Günther

Abstract. Fry of Cichlasoma urophthalmus Günther were exposed to various concentrations of the organophosphate pesticide Trichlorfon, ranging from 1·25 to 80 mg/l in laboratory conditions. Mortality reached 100% in the highest concentrations (60 and 80 mg/l) in the first 24h. The intermediate concentration (40 mg/l) resulted in 50% mortality after 24h, and no mortality was observed in lower levels of the pesticide. LC50 at 24, 48 and 72h were 26·5, 23·7 and 17·2 mg/l respectively.
Some of the surviving fish were transferred to clean fresh water for a post-treatment growth trial. All the fish grew well with no apparent adverse effects as judged by their growth indices after 8 weeks.     

Changes in selected stress and immune‐related genes in Atlantic cod,Gadus morhua,following overcrowding

Using Atlantic cod, Gadus morhua, as a model for the stress response in gadoid fish, the changes in the expression of some stress and immune genes as well as the profiles of plasma cortisol were examined. Adult fish were kept at a density of ca. 100 kg m^?3 by lowering the water level in the rearing tank for 1 h and this short‐term crowding stress was repeated thrice over a 12‐h interval period. Blood samples were collected before exposure and at 2, 24 and 72 h post crowding. Plasma cortisol level significantly increased at 2 h post crowding but returned to pre‐crowding levels 24 h after exposure. The relative expression of the stress response genes, glucose transporter‐3 and a putative heat shock protein 70 significantly increased at 2 and 24 h post crowding respectively. Significant up‐regulation of the pro‐inflammatory cytokines, interleukin (IL)‐1β and IL‐8, as well as anti‐bacterial genes, g‐type lysozyme and bactericidal permeability‐increasing protein/lipopolysaccharide‐binding protein (BPI/LBP) was also observed at 2 h and the levels were maintained until 72 h post exposure, except for BPI/LBP which had maximum up‐regulation at 24 h. The present observations have implications with respect to fish welfare and assessment of the health status of the farmed fish.     

River pollution from a large trout farm in Norway

The pollution loading from Øxna Bruk, a trout farm with an annual fish production of 200–250 tons, and the influence on the water quality of River Figgjo, a mediumsized watercourse, was studied. The feed used was dry pellets. The loadings were estimated to be 0.3–0.8 g N/kg fish.24h; 0.05 g P/kg fish.24h; and 1.6–4.6 g O/kg fish.24 h as BOD₇ (biochemical oxygen demand, 7 days). As a result of the fish farming activities, moderate chemical alterations were registered in the river downstream of the farm at low river flow: decreased oxygen concentration (÷ 1 mg/l) and increased concentrations of total-N (+ 0.2 mg N/l), NH₃ + NH₄⁺ (+ 0.1 mg N/l), total-P (+ 0.03 mg P/l) and ortho-P (+ 0.02 mg P/l). Compared with the receiving water, BOD₇ increased maximally 1 mg O/l downstream of the farm. The salmonid stocks in the river were not affected by the effluents from the trout farm.