The interactive effect of elevated temperature on deltamethrin-induced biochemical stress responses in Channa punctata Bloch

There are reports showing interactive effect of environmental factors with the toxic outcome of chemicals. We studied the interactive effect of elevated temperature as an abiotic stressor on deltamethrin-induced biochemical stress responses in a freshwater fish, Channa punctata Bloch. Heat stress (∼12 °C above ambient temperature for 3 h) and pesticide exposure (deltamethrin 0.75 ppb for 48 h) showed significant induction of heat shock protein-70 (HSP70) in liver, kidney and gills of fishes. Elevated temperature when followed by deltamethrin exposure showed synergistic effect showing a high level of HSP70 in liver and gills whereas response in the kidney was opposite. On the contrary, when deltamethrin exposure followed the heat stress, no significant difference was observed. Protein carbonylation was found to be more pronounced in heat-stressed group compared with control fish group. A significant increase in lipid peroxidation (LPO) was observed in different tissues of fish exposed to either of the stressors. In the kidney of fish exposed to heat stress followed by deltamethrin, LPO was relatively lower as compared to other treatments. Thiols content such as reduced glutathione (GSH), total thiols (T-SH), non-protein thiols (NP-SH) and protein thiols (P-SH) showed no consistent pattern in different tissues. In deltamethrin-exposed group that was subsequently exposed to heat stress, the GSH content was higher in liver and lower in both kidney and gills when compared with other groups. Alteration in the activities of antioxidant enzymes such as catalase (CAT), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) was also observed when fish were exposed to heat stress and/or deltamethrin. Our study demonstrated that heat stress modulated biochemical stress responses in fish showing a tissue specific pattern. This implies that fish has the capacity to elicit differential response to exposure to abiotic stressors in order to reduce the systemic magnitude of stress which may otherwise lead to severe dysfunction of vital tissues.     

Short‐term freshwater exposure benefits sea lice‐infected Atlantic salmon

Atlantic salmon Salmo salar were infected with sea lice Lepeophtheirus salmonis (0·08 ± 0·007 sea lice g⁻¹) over a period of 4 h. Both infected and non‐infected fish were swim tested in sea water (SW) and fresh water (FW). The ventral aorta of each fish was fitted with a Doppler cuff in order to measure cardiac output, stroke volume and heart rate during swim testing. Blood samples were taken at rest and after exercise. Critical swimming speed of infected fish in SW (2·14 ± 0·08 body lengths, bl s⁻¹) was significantly lower ( P  < 0·05) than infected fish switched to FW (2·81 ± 0·08 bl s⁻¹) and non‐infected fish in SW (2·42 ± 0·04 bl s⁻¹) and FW (2·61 ± 0·08 bl s⁻¹). Cardiac and blood results indicated infected fish exposed to FW did experience stress, but relief from osmotic and ionic distress probably reduces energy expenditure, allowing the increase in performance. As the performance of sea lice‐infected fish improved upon transfer to FW, it is likely that heavily infected salmonids do return to FW to restore compromised osmotic and ionic balance, and remove sea lice in the process.     

Influence of temperature on silver accumulation and depuration in rainbow trout

To assess the influence of water temperature on silver uptake, rainbow trout Oncorhynchus mykiss ( c . 50 g; held at 13° C) were exposed to 0·1 μM AgNO₃ in ion‐poor water for 1 week at 4 and 16° C without previous temperature acclimation. To assess the influence of temperature on elimination of previously accumulated Ag, rainbow trout were exposed to 0·1 μM AgNO₃ in ion‐poor water for 1 week at 12° C, then were randomly divided amongst two Ag‐free water containers, differing only in temperature (3 and 16° C), for 2 months. In the uptake study greater accumulation of Ag was seen in the gills, plasma and especially the livers and bile of 'warm' rainbow trout (16° C) compared to 'cold' rainbow trout (4° C), which can be explained by the higher metabolic rates of the warmer fish. In the depuration study there was no net elimination of Ag from the livers and bile but there was biphasic elimination of Ag from the gills and plasma of 'warm' and 'cold' fish, but with few differences between them. This indicated that temperature‐dependent processes were less important in Ag elimination than in Ag uptake. Toxicokinetic modelling of Ag uptake by livers indicated four‐fold greater uptake of Ag by 'warm' rainbow trout compared to 'cold' rainbow trout (one compartment uptake model). Elimination of previously accumulated Ag from the plasma was best fitted by a two compartment rate‐constant based model, with approximately half the plasma Ag load eliminated within 24 h, followed by slower elimination of Ag over 2 months.     

Swimming activity of seabass: comparing patterns obtained in natural environment and in re-circulating tanks under high density

Seabass ( Dicentrarchus labrax ) swimming activity was compared between natural environments and aquaculture facilities. Behaviour under natural conditions was assessed in a saltmarsh pond (250 m², 18 × 14 × 0·8 m) using acoustic telemetry. From several surveys, we documented the diel activity rhythm and demonstrated group effects on swimming patterns and amplitudes by comparing activity of solitary fish with that of a fish living in a group of 60. Consequences of weather variability were also analysed and revealed a high sensitivity of fish to atmospheric conditions for both swimming and demand‐feeding behaviour. Behaviour in fish tanks was also studied using acoustic telemetry, as part of the EUREKA EU1 960 'Aqua‐Maki 2' project investigating aspects of fish culture in re-circulating tanks under high density. A re-circulating hexagonal tank (5·4 × 5·4 m, 1·8 m depth, 48 m³) was equipped with positioning and demand‐feeding systems, oxygen and temperature probes. Initial density was 50 kg m³ in March and rose to 90 kg m³ at the end of the experiment in May. During this period, the movements of nine fish were continuously recorded for 24 h each, reaching a total of six 24 h episode at eight days interval. Swimming activity was analysed in terms of activity rhythms and space occupation specially around feeding events. The two data set and main results will be presented and compared to assess seabass behavioural plasticity and sensitivity to husbandry conditions.     

Effects of waterborne complexing agents on silver uptake and depuration in rainbow trout

Rainbow trout Oncorhynchus mykiss ( c . 60 g) were exposed for 1 week to 0·1 μM silver as AgNO₃ in ion poor water (Ca c . 150 μM, pH c . 8, water temperature 13° C) with or without waterborne organic matter (27 mg C l⁻¹ as Aldrich humic acid), thiosulphate (5 μM Na₂S₂O₃) or chloride (4 mM KCl). Organic matter decreased Ag accumulation by the gills initially, but did not decrease Ag accumulation by plasma or liver. Thiosulphate decreased the amount of Ag accumulated by the gills for the entire 1 week exposure but had no effect on Ag concentrations in the plasma, liver or bile. Chloride had no effect on Ag uptake in any of the tissues examined. All three complexing agents reduced the decreases in plasma Na and Cl concentrations caused by Ag. To study the effects of waterborne complexing agents on Ag depuration, rainbow trout were exposed to 0·1 μM AgNO₃ for 1 week then placed for 8 days in Ag‐free, ion poor water with or without waterborne organic matter (55 mg C l⁻¹) or thiosulphate (5 μM). These complexing agents did not alter depuration of Ag from the gills, plasma, liver or bile. Thus, once Ag has entered a fish, subsequent elimination of internal Ag is not affected by external complexing agents.     

Influence of dietary protein level on growth performance and body composition of juvenile blackspot seabream, Pagellus bogaraveo(Brunnich, 1768)

Blackspot seabream Pagellus bogaraveo is a potential candidate species for southern European aquaculture. A 12‐week feeding trial was then conducted to evaluate the effect of dietary protein level on growth performance, body composition and feed use efficiency of blackspot seabream juveniles (22·7 g each). Five isolipidic diets (12·5%) with graded levels of protein (20, 30, 40, 50 and 60%) were hand‐distributed, twice a day, to duplicate groups of fish (120 fish per tank), until satiation. Growth (DGC), survival and feed utilization (FCR) were recorded through monthly sampling. Growth performance was significantly affected by dietary protein level ( P  < 0·05), and fish fed at the 50% protein level exhibited the highest final body mass. Feed conversion ratio decreased with increasing levels of dietary protein, fish fed 60% protein being the most efficient. No significant differences were found in whole body composition among treatments ( P > 0·05). Financed by FCT (Project POCTI/CVT/39239/2001, PhD Grant SFRH‐BD‐14068–2003)     

Finlets and the steady swimming performance of Thunnus albacares

The functional significance of finlets on the steady swimming performance of yellowfin tuna Thunnus albacares was evaluated by measuring the speed and tail‐beat frequency of the fish with and without them. It was hypothesized that if finlets do improve swimming performance, fish without finlets would have to work harder to maintain the same swimming speed as fish with them and that this would be reflected in kinematic differences. Two‐way ANOVA showed significant effects between individuals on speed (d.f. = 5 and 228, P  < 0·001) and tail‐beat frequency (d.f. = 5 and 48, P  < 0·001), but no significant effects of treatment on speed (d.f. = 1 and 228, P  = 0·25) and tail‐beat frequency (d.f. = 1 and 48, P  > 0·1). No interaction effects on speed (d.f. = 5 and 228, P  > 0·1) and tail‐beat frequency (d.f. = 5 and 48, P  > 0·25) were found. This suggested that finlets were unlikely to function as significant drag reduction and thrust enhancing devices in routine steady swimming. Though not statistically significant, small percentage differences between the mean swimming speeds and tail‐beat frequency of the untreated and treated groups (fish with and without finlets respectively) of the order of 0·5% may be meaningful over the life of a fish. Also, finlets may improve performance at high sustained speeds in rapid accelerations and turns.     

Environmental stress and bacterial infection in channel catfish, Ictalurus punctatus Rafinesque

Channel catfish, Ictalurus punctatus , were injected intraperitoneally with a sublethal dose of Aerornonas hydrophila and then stressed for 144 h by being maintained either in a dissolved oxygen concentration of 1·5 mg/1, 1·2 mg/1 total ammonia, and/or 6·5 mg/1 free CO₂ with a continuous inflow of water. A significant difference in percentage of mortality was noted between treatments ( P < 0·05). The trunk kidneys of surviving stressed fish had significantly higher total bacterial counts than non-stressed controls. A. hydrophila was isolated from 67% of the stressed fish and 9% of the control fish. Edwardsiella tarda , apparently endemic in the population, was isolated from 43% of the stressed fish and 7% of the control fish. Histopathological lesions were in the gills, liver, spleen, trunk kidney, and head kidney of stressed fish, but not control fish.     

Ontogenetic changes in temperature preference of Atlantic cod

Final thermal preferendum ( T ) experiments were conducted in a horizontal thermal gradient tank from the beginning of August 2001 to mid‐November 2001 using Atlantic cod Gadus morhua from 6·5 to 79·0 cm fork length ( L _F). The value of T varied significantly ( P  < 0·005) with L _F( T  = 7·23–0·054 L _F), with smaller (younger) fish choosing higher temperatures than larger (older) fish. The preferendum varied from 6·9° C for fish of 6·5 cm to 3·0° C for those of 79·0 cm. Experiments comparing fish positions in the gradient tank between thermal gradients of 0·5–11·0 and 4·5–14·5° C demonstrated that fish positions were determined by temperature selection instead of undesirable tank effects. This study is the first to demonstrate the effect of ontogeny on temperature preferences of a marine fish species.     

The effect of temperature and acclimation period on repeat swimming performance in cutthroat trout

Hatchery cutthroat trout Oncorhynchus clarki clarki were used to examine the effects of 48 h and 3 week temperature acclimation periods on critical swimming speed ( U _crit). The U _crit was determined for fish at acclimation temperatures of 7, 14 and 18° C using two consecutive ramp‐ U _crit tests in mobile Brett‐type swim tunnels. An additional group was tested at the stock's ambient rearing temperature of 10° C. The length of the temperature acclimation period had no significant effect on either the first or the second U _crit( U _crit‐1 and U _crit‐2, respectively) or on the recovery ratio (the quotient of U _crit‐2  U _crit‐1⁻¹). As anticipated, there was a significant positive relationship between U _crit‐1 and temperature ( P  < 0·01) for both acclimation periods, and an increasing, though non‐significant, trend between U _crit‐2 and temperature ( P  = 0·10). Acclimation temperature had no significant effect ( P  = 0·71) on the recovery ratio. These results indicate that a 48 h acclimation to experimental temperatures within the range of −3 to +8° C of the acclimation temperature may be sufficient in studies of swimming performance with this species. This ability to acclimate rapidly is probably adaptive for cutthroat trout and other species that occupy thermally variable environments.     

Influence of rearing temperature during the larval and nursery periods on growth and sex differentiation in two Mediterranean strains of Dicentrarchus labrax

European sea bass Dicentrarchus labrax of the north‐western (NW) and south‐eastern (SE) Mediterranean Sea strains were exposed to different temperatures (13, 17 or 21° C) during the larval rearing (11–51 days post hatching, dph) or nursery periods (55–95 dph), in order to examine the effects of temperature on sex differentiation and subsequent growth during the first year of life. Higher growth was observed during exposure to higher temperatures, but fish of the NW strain exposed to 13 or 17° C during larval rearing exhibited compensatory growth once exposure to the lower temperatures finished, and as a result their final size at 300 dph was similar or greater to the group exposed to 21° C. Fish exposed to 17° C during the nursery period also had similar size to fish exposed to 21° C after 300 days of rearing, but the fish exposed to 13° C remained significantly smaller (ANOVA, n  = 55–100, P  < 0·05). There were significant differences in the sex ratio among the fish exposed to different temperatures during the two periods of rearing, with high temperature (21° C) resulting in a significantly higher percentage of males in the population, both in the NW (ANOVA, n  = 2, P  < 0·04) and SE populations (ANOVA, n  = 2, P  < 0·01). The masculinization effect of high temperature was significantly stronger during the larval rearing stage, both in the NW (ANOVA, n  = 2, P  < 0·005) and SE populations (ANOVA, n  = 2, P  < 0·01). None of the temperature manipulations could produce 100% females, suggesting that there is a part of the genetic component in sex differentiation which is not labile to environmental influence.     

A field and experimental evaluation of the effect of data storage tags on the growth of cod

The effect of electronic data storage tags (DSTs) on the growth of cod Gadus morhua was evaluated in laboratory and field experiments. In the first laboratory experiment, large DSTs (60 × 18 mm, 3 g in water ) attached externally for 3 months did not have any effect ( anova , P  > 0·05) on the growth of adult cod (mean ±  s . d . 65 ± 4·5 cm total length) relative to untagged adult cod. In a second experiment, small DSTs (34 × 11 mm, 1·5 g in water) implanted into young cod (48·1 ± 4·4 cm) for an 8 month period did not have any effect upon the growth relative to untagged controls ( anova , P  > 0·05). Length data returned from tagging experiments conducted on adult cod (57·3 ± 7·5 cm) in the North Sea showed that the growth of fish tagged either externally or internally with large DSTs was not different ( t ‐test, P  > 0·05). Attachment wounds, however, provided evidence that external attachment of DSTs should be avoided unless sensor configuration requires access to the external environment, and that internal implantation should be preferred whenever possible.     

A new technique to gather 3‐D spatial information using a single camera

A new technique using a single camera and shadows to determine 3‐D spatial positions of fishes in the laboratory is described. The apparatus consisted of a large aquarium (2·0 × 1·5 × 0·4 m), a wide‐angle camera mounted above and two light sources to cast shadows to either side of the fish. Using image analysis and vector mathematics, aquarium objects were plotted within 1·5 cm of their actual location along the x ‐, y ‐ and z ‐axis. The technique was also successful in quantifying changes in 3‐D spatial pattern of juvenile fish, Atlantic cod Gadus morhua (7·4–8·6 cm standard length, L _S) and cohabitant piscivorous shorthorned sculpin Myoxocephalus scorpinus (12·0–25·8 cm L _S), at these same viewing fields. The new technique should have a wide application, largely because it is potentially less expensive, laborious and invasive than alternative methods for determining 3‐D positions of fishes.     

Evaluation of in vitro endocytosis and antibody synthesis by rainbow trout head kidney cells treated with bovine lactoferrin

Bovine lactoferrin (LF) was evaluated for its capacity to modulate the in vitro endocytosis (phagocytosis and pinocytosis) and antibody synthesis by head kidney cells of rainbow trout Oncorhynchus mykiss . Phagocytic activity and phagocytic index of head kidney macrophages, determined by measurement of ingested yeast, were influenced by bovine LF starting from the LF concentration of 1 and 0·1 μg ml⁻¹, respectively. Endocytosis, determined by the evaluation of droplet uptake of neutral red dye solution, was significantly enhanced by 10 μg ml⁻¹ of LF. In contrast, antibody synthesis by head kidney cells, evaluated by immunoenzymatic assay, from fish immunized against human‐γ‐globulins (HGG) in vivo was not affected by bovine LF. Although these results showed that bovine LF had no effect on specific immunoglobulin production in vitro , an enhancement of the acquired immune response may be assumed in LF‐treated fish in vivo , as observed in higher vertebrates.     

Lipid content and fatty acid composition of target tissues in wild Perca fluviatilis females in relation to hepatic status and gonad maturation

Variation in liver ultra‐structure and composition in relation to energy mobilization was investigated in female perch Perca fluviatilis from the Meuse River between August 2001 and June 2002. In April, just before spawning, the lipo‐somatic index ( I _F) was 0·3%, the gonado‐somatic index ( I _G) was 28% and the total lipid content of the liver was 2·53%. The average areas of lipid droplets and mitochondria were 0·05 and 0·06 μm², respectively. Glycogen supply reached 7·9% of the total area of the hepatocyte. During the sexual resting period, females accumulated energy in perivisceral fat and in the liver to reach 1·6% I _F and 4·85% of liver lipid content in August with lipid droplets average size of 0·09 μm² and glycogen average area of 15%. Liver cells contained a weakly developed rough endoplasmic reticulum (RER) and a great number of small mitochondria (average size 0·02 μm²). The I _G was 0·6% at this time. During the whole annual cycle, the average lipid content of female liver never exceeded 3·9 ± 1·9%. The concentration of docosahexaenoic (DHA), linolenic and linoleic acids increased in mature gonads while linolenic and linoleic acids decreased in the liver during the same period. Fatty acid composition of muscles of perch was characterized by a high content of DHA.     

Is rigor mortis the cause of post‐mortem shrinkage in juvenile Pagrus auratus?

An experiment demonstrated that post‐mortem shrinkage in fork length of juvenile pink snapper Pagrus auratus (179–262 mm) was the same in fish in which rigor mortis was surgically inhibited, as in intact fish, and averaged 5·5 ± 0·2 mm (mean ±  s.e ., range 2–9 mm) after 168 h.     

Feeding preference and deterrence in rabbitfish Siganus fuscescens for the cyanobacterium Lyngbya majuscula in Moreton Bay, south‐east Queensland, Australia

Rabbitfish Siganus fuscescens preferences for Lyngbya majuscula collected from three bloom locations in Moreton Bay, Queensland, Australia, were tested along with a range of local plant species in the laboratory. Consumption of L. majuscula by fish did not differ between wild and captive‐bred fish ( P  = 0·152) but did differ between bloom location ( P  = 0·039). No relationship was found between consumption rates and lyngbyatoxin‐a concentration ( r ² = 0·035, P  = 0·814). No correlation existed between C : N and proportion of food consumed when all food types were analysed statistically, whereas a clear correlation was observed when L. majuscula was removed from the calculations. In simulated bloom conditions, fish avoided ingestion of L. majuscula by feeding through gaps in the L. majuscula coverage. Both wild and captive‐bred S. fuscescens showed a distinct feeding pattern in 10 day no‐choice feeding assays, with less L. majuscula being consumed than the preferred red alga Acanthophora spicifera . Lyngbya majuscula however, was consumed in equal quantities to A. spicifera by wild S. fuscescens when lyngbyatoxin‐a was not detectable. Wild fish probably do not preferentially feed on L. majuscula when secondary metabolites are present and are not severely impacted by large L. majuscula blooms in Moreton Bay. Furthermore, poor feeding performance in both captive‐bred and wild S. fuscescens suggests that they would exert little pressure as a top‐down control agent of toxic L. majuscula blooms within Moreton Bay.     

Inter‐individual differences in rates of routine energy loss and growth in young‐of‐the‐year juvenile Atlantic cod

Inter‐individual differences in rates of routine (non‐feeding) metabolism and growth were evaluated in young‐of‐the‐year (YOY) juvenile Atlantic cod Gadus morhua . Rates of O₂ consumption, CO₂ production and ammonia (TAN) excretion were measured in 64, 25–43 mm standard length ( L _S) YOY growing at different rates (0·27–0·47 mm day⁻¹) in a common rearing tank. Parameter rates ( y ) increased allometrically ( y = a·M^b ) with increasing body mass ( M ) with b ‐values for O₂ production, CO₂ consumption and TAN excretion equal to 0·81, 0·89 and 0·56, respectively. In some cases, residuals from these regressions were significantly negatively correlated to fish growth rate. In no cases did residuals of parameter rates increase with increasing growth rate. These data suggest that, during unfed periods, relatively fast‐growing fish were more metabolically efficient than slower‐growing fish from the same cohort. The fish condition factor, derived from     , also significantly decreased with increasing growth rate. Results indicated differences in both the rates of routine energy loss and the patterns of growth allocation among YOY Atlantic cod. Since these physiological attributes were positively correlated with growth rate, they may be indicative of 'survivors' in field populations.     

Light intensity, prey detection and foraging mechanisms of age 0 year yellow perch

The ability of age‐0 year yellow perch Perca flavescans to detect prey using visual and mechano‐sensory input was examined during laboratory feeding trials at varying light intensities. Perch were highly effective predators and captured Daphnia pulicaria with 94% overall foraging success at light levels ranging from 0 to 3400 lx. Maximum average reaction distances (5·0 ± 0·8 cm, mean ±  s . e .) occurred in front of the fish at 3000 lx and significantly decreased as light intensities fell to <2 lx, with minimum reaction distances (2·8 ± 0·1 cm) observed in the dark. Following chemical ablation of the lateral line, yellow perch showed a significant reduction in reaction distance when compared to the untreated fish at 3000 lx, suggesting that the lateral line may augment visual prey detection at high light levels. A model was created to predict reaction distances for fish feeding with multiple sensory systems that can be applied to a variety of photic environments. This study provides a better understanding of the contribution of vision and the lateral line to prey detection, and relates the reaction distance of age‐0 year yellow perch to light intensities similar to those experienced in nature.     

The effects of selenium on oxidative stress biomarkers in the freshwater characid fish matrinxã, Brycon cephalus (Günther, 1869) exposed to organophosphate insecticide Folisuper 600 BR (methyl parathion)

Methyl parathion (MP), an organophosphate widely applied in agriculture and aquaculture, induces oxidative stress due to free radical generation and changes in the antioxidant defense system. The antioxidant roles of selenium (Se) were evaluated in Brycon cephalus exposed to 2 mg L(-1) of Folisuper 600 BR (MP commercial formulation - MPc, 600 g L(-1)) for 96 h. Catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione S-transferase (GST), reduced glutathione (GSH) and lipid peroxidation (LPO) levels in the gills, white muscle and liver were evaluated in fish fed on diets containing 0 or 1.5 mg Se kg(-1) for 8 weeks. In fish treated with a Se-free diet, the MPc exposure increased SOD and CAT activities in all tissues. However, the GPx activity decreased in white muscle and gills whereas no alterations were observed in the liver. MPc also increased GST activity in all tissues with a concurrent decrease in GSH levels. LPO values increased in white muscle and gills and did not change in liver after MPc exposure. A Se-supplemented diet reversed these findings, preventing increases in LPO levels and concurrent decreases in GPx activity in gills and white muscle. Similarly, GSH levels were maintained in all tissue after MPc exposure. These results suggest that dietary Se supplementation protects cells against MPc-induced oxidative stress.