Experimental evaluation of using calcein and alizarin red S for immersion marking of bighead carp Aristichthys nobilis (Richardson, 1845) to assess growth and identification of marks in otoliths,scales and fin rays

In order to evaluate the effects of immersion marking with calcein (CAL) and alizarin red S (ARS) on growth and mortality of juvenile bighead carp Aristichthys nobilis, and assess mark quality in otoliths, scales, and fin rays, CAL from 50 to 200 mg L^?1 and ARS from 150 to 300 mg L^?1 concentrations were used. With the exception of non‐lateral line scales from 50 mg L^?1 CAL treatments, immersion for 24 h produced detectable marks in sagittae, lateral line and non‐lateral line scales, and fin rays (dorsal, pectoral, ventral, anal, and caudal) at 100 days post‐marking. Detectable fluorescent marks in sagittae were readily observed at concentrations of 150–200 mg L^?1 CAL or 150–300 mg L^?1 ARS. Marks were poorly visible in all non‐lateral line scales from both CAL‐ and ARS‐treated groups. Fluorescent marks were readily detected in lateral line scales at 100–200 mg L^?1 CAL or 150–300 mg L^?1 ARS, and in fin rays at 150–200 mg L^?1 CAL or 150–300 mg L^?1 ARS. In particular, optimal marks were observed at the highest concentrations investigated in sagittae (300 mg L^?1 ARS), lateral line scales (150–200 mg L^?1 CAL or 250–300 mg L^?1 ARS), and fin rays (200 mg L^?1 CAL or 250–300 mg L^?1 ARS). However, fluorescent marks visible to the naked eye were not produced by any of the CAL or ARS treatments in sagittae, scales, or fin rays during this experiment. In addition, there was no significant difference on survival and growth of marked fish compared to controls throughout the experiment (P > 0.05).     

Trace element and strontium isotopic analysis of Gulf Sturgeon fin rays to assess habitat use

Trace element and ⁸⁷Sr/⁸⁶Sr isotope analyses of fish pectoral fin rays offer non-destructive methods for determining habitat use. In this study, water and fin ray samples were analyzed for Gulf Sturgeon Acipenser oxyrinchus desotoi from the Choctawhatchee River Basin (FL and AL, USA) and compared with reference samples from Atlantic Sturgeon A. o. oxyrinchus held at controlled salinities (0, 10, 33 ppt). Samples were analyzed using inductively coupled plasma mass spectrometry, with a multi-collector for ⁸⁷Sr/⁸⁶Sr. In water, Sr, Ba, Mn and Zn differed between freshwater and saline habitats, with increases in Sr and decreases in Ba, Mn and Zn. ⁸⁷Sr/⁸⁶Sr decreased upstream to downstream with lowest values in saline habitats. In the reference study, water trace element concentrations and ⁸⁷Sr/⁸⁶Sr corresponded to those in pectoral fin rays. ⁸⁷Sr/⁸⁶Sr was higher in pectoral fin ray than water, due to influence of diet, which differed with salinity. In wild fish, trace elements in pectoral fin rays indicated freshwater emigration to saline habitats primarily occurred in the second to third growth zone with some heterogeneity in the population (4% <0.3 years, 39% 0.5–1.3 years, 39% 1.5–2.3 years, 17% 2.5–3.3 years). Analyses of ⁸⁷Sr/⁸⁶Sr indicated initial locations of Gulf Sturgeon were in the middle river, with few fish in the upper or lower river. Most (74%) juvenile Gulf Sturgeon utilized more than one river region prior to freshwater emigration and 48% moved upstream temporarily based on increased ⁸⁷Sr/⁸⁶Sr. After initial freshwater emigration, fish utilized lower-river to saline habitats. Collectively, these studies demonstrate the usefulness of trace element and ⁸⁷Sr/⁸⁶Sr analyses in sturgeon pectoral fin rays.     

Mass‐marking of farmed European eels (Anguilla anguilla (Linnaeus, 1758)) with alizarin red S

The Working Group on Eel of the International Council for the Exploration of the Sea (ICES) regularly reports that a significant amount of stocked eels in Europe was pre‐grown in aquaculture farms prior to stocking—so called “farmed eels.” The ICES advices chemical marking of stocked recruits to ensure their traceability throughout all life stages. To date, however, there was a lack of knowledge concerning the most suitable chemical substance and its application on farmed eels. The aim of this study was to fill this gap by presenting successful attempts of marking those eels with alizarin red S (ARS). An ARS concentration of 150 mg L^?1 buffered with 150 mg L^?1 Tris(hydroxymethyl)aminomethane applied as an immersion bath over 9 h was sufficient to mark a total of 3572 kg of farmed eels (6.5–8.0 g mean body weight). The marking success was 100% on otoliths and highest stocking density of up to 67.1 kg m^?3 (corresponding 54.0 kg m^?2) turned out to have no effect on mortality which was consistently below 1%.     

Functional morphology of the fin rays of teleost fishes

Ray‐finned fishes are notable for having flexible fins that allow for the control of fluid forces. A number of studies have addressed the muscular control, kinematics, and hydrodynamics of flexible fins, but little work has investigated just how flexible ray‐finned fish fin rays are, and how flexibility affects their response to environmental perturbations. Analysis of pectoral fin rays of bluegill sunfish showed that the more proximal portion of the fin ray is unsegmented while the distal 60% of the fin ray is segmented. We examined the range of motion and curvatures of the pectoral fin rays of bluegill sunfish during steady swimming, turning maneuvers, and hovering behaviors and during a vortex perturbation impacting the fin during the fin beat. Under normal swimming conditions, curvatures did not exceed 0.029 mm^?1 in the proximal, unsegmented portion of the fin ray and 0.065 mm^?1 in the distal, segmented portion of the fin ray. When perturbed by a vortex jet traveling at approximately 1 ms^?1 (67 ± 2.3 mN s.e. of force at impact), the fin ray underwent a maximum curvature of 9.38 mm^?1. Buckling of the fin ray was constrained to the area of impact and did not disrupt the motion of the pectoral fin during swimming. Flexural stiffness of the fin ray was calculated to be 565 × 10^?6 Nm². In computational fluid dynamic simulations of the fin‐vortex interaction, very flexible fin rays showed a combination of attraction and repulsion to impacting vortex dipoles. Due to their small bending rigidity (or flexural stiffness), impacting vortices transferred little force to the fin ray. Conversely, stiffer fin rays experienced rapid small‐amplitude oscillations from vortex impacts, with large impact forces all along the length of the fin ray. Segmentation is a key design feature of ray‐finned fish fin rays, and may serve as a means of making a flexible fin ray out of a rigid material (bone). This flexibility may offer intrinsic damping of environmental fluid perturbations encountered by swimming fish. J. Morphol. 274:1044–1059, 2013. © 2013 Wiley Periodicals, Inc.     

Tagging the next generation: validation of trans-generational chemical tagging for an endangered fish

In this study we validated marking offspring through peritoneal injection of ripe females using two concentrations of strontium (strontium chloride hexahydrate). Larvae from treatments were monitored for condition morphometrics and tested for chemical mark incorporation in their otoliths via laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) to quantify the strontium concentration (Sr/Ca) and laser ablation multi-collector inductively coupled plasma mass spectrometry (LA-MC-ICPMS) to measure the strontium isotope ratios (⁸⁷Sr:⁸⁶Sr) of otoliths. Otolith strontium concentrations and strontium isotopes ratios were elevated in the high-concentration treatment, while the low-concentration and control treatments were not significantly different from each other. Larval size and eye diameter at hatch were similar among treatments; however, yolk and oil globule diameters were significantly reduced in the high-concentration treatment. Moreover, growth rates after 60?days post-hatch were significantly reduced in the high-concentration treatment relative to the low-concentration and control treatments, suggesting trans-generational tagging can have deleterious effects on offspring. Our study provides evidence for the efficacy of artificially marking offspring via injection of strontium into ripe females and could provide new tools for managing endangered fish populations; however, careful consideration of chemical concentrations and dosages may be required prior to its application in a fisheries management experiment.     

Composition of the early Oligocene ocean from coral stable isotope and elemental chemistry

A sectioned and polished specimen of the coral Archohelia vicksburgensis from the early Oligocene Byram Formation (～30 Ma) near Vicksburg, Mississippi, reveals 12 prominent annual growth bands. Stable oxygen isotopic compositions of 77 growth‐band‐parallel microsamples of original aragonite exhibit well‐constrained fluctuations that range between ?2.0 and ?4.8. Variation in δ¹⁸O of coral carbonate reflects seasonal variation in temperature ranging from 12 to 24 °C about a mean of 18 °C. These values are consistent with those derived from a bivalve and a fish otolith from the same unit, each using independently derived palaeotemperature equations. Mg/Ca and Sr/Ca ratios were determined for 40 additional samples spanning five of the 12 annual bands. Palaeotemperatures calculated using elemental‐ratio thermometers calibrated on modern corals are consistently lower; mean temperature from Mg/Ca ratios are 12.5 ± 1 °C while those from Sr/Ca are 5.8 ± 2.2 °C. Assuming that δ¹⁸O‐derived temperatures are correct, relationships between temperature and elemental ratio for corals growing in today's ocean can be used to estimate Oligocene palaeoseawater Mg/Ca and Sr/Ca ratios. Calculations indicate that early Oligocene seawater Mg/Ca was ～81% (4.2 mol mol^?1) and Sr/Ca ～109% (9.9 mmol mol^?1) of modern values. Oligocene seawater with this degree of Mg depletion and Sr enrichment is in good agreement with that expected during the Palaeogene transition from ‘calcite’ to ‘aragonite’ seas. Lower Oligocene Mg/Ca probably reflects a decrease toward the present day in sea‐floor hydrothermal activity and concomitant decrease in scavenging of magnesium from seawater. Elevated Sr/Ca ratio may record lesser amounts of Oligocene aragonite precipitation and a correspondingly lower flux of strontium into the sedimentary carbonate reservoir than today.     

Production rate estimation of mycosporine‐like amino acids in two Arctic melt ponds by stable isotope probing with NAH13CO3

The net carbon uptake rate and net production rate of mycosporine‐like amino acids (MAAs) were measured in phytoplankton from 2 different melt ponds (MPs; closed and open type pond) in the western Arctic Ocean using a ¹³C stable isotope tracer technique. The Research Vessel Araon visited ice‐covered western‐central basins situated at 82°N and 173°E in the summer of 2012, when Arctic sea ice declined to a record minimum. The average net carbon uptake rate of the phytoplankton in polycarbonate (PC) bottles in the closed MP was 3.24 mg C · m^?3 · h^?1 (SD = ±1.12 mg C · m^?3 · h^?1), while that in the open MP was 1.3 mg C · m^?3 · h^?1 (SD = ±0.05 mg C · m^?3 · h^?1). The net production rate of total MAAs in incubated PC bottles was highest (1.44 (SD = ±0.24) ng C · L^?1 · h^?1) in the open MP and lowest (0.05 (SD = ±0.003) ng C · L^?1 · h^?1) in the closed MP. The net production rate of shinorine and palythine in incubated PC bottles at the open MP presented significantly high values 0.76 (SD = ±0.12) ng C · L^?1 · h^?1and 0.53 (SD = ±0.06) ng C · L^?1 · h^?1. Our results showed that high net production rate of MAAs in the open MP was enhanced by a combination of osmotic and UVR stress and that in situ net production rates of individual MAA can be determined using ¹³C tracer in MPs in Arctic sea ice.     

Chemical marking of European glass eels Anguilla anguilla with alizarin red S and in combination with strontium: in situ evaluation of short‐term salinity effects on survival and efficient mass‐marking

This study presents a new chemical double‐marking technique for European glass eels Anguilla anguilla by combing alizarin red S (ARS) and strontium chloride hexahydrate (Sr). Marked eels (double marked with ARS and Sr, but also single marked with ARS) were exposed in situ to brackish water (15 g l^?1 artificial sea salt) for 14 days and did not exhibit increased mortalities compared with unmarked eels. Indeed, no mortality occurred in a marked group during the experiments. Moreover, an efficient mass‐marking approach with low handling effort for both single ARS and double ARS‐Sr techniques is described and was proven to be practicable for large‐scale stocking programmes.     

Comparison of fin ray sampling methods on white sturgeon Acipenser transmontanus growth and swimming performance

Effects of two fin‐ray sampling methods on swimming performance, growth and survival were evaluated for hatchery‐reared sub‐adult white sturgeon Acipenser transmontanus. Fish were subjected to either a notch removal treatment in which a small section was removed from an anterior marginal pectoral‐fin ray, or a full removal treatment in which an entire marginal pectoral‐fin ray was removed. Control fish did not have fin rays removed, but they were subjected to a sham operation. A modified 3230 l Brett‐type swim tunnel was used to evaluate 10 min critical station‐holding speeds (S_CSH) of A. transmontanus, immediately after the fin ray biopsies were obtained with each method. Survival and growth were evaluated over a 6 month period for a separate group of fish subjected to the same biopsy methods. Mean ± s.e . 10 min S_CSH were 108·0 ± 2·3, 110·0 ± 2·6 and 115·0 ± 3·5 cm s^?1 for the notch removal group, full removal group and control group, respectively, and were not significantly different among treatments. Behavioural characteristics including tail‐beat frequency and time spent hunkering were also not significantly different among treatment groups swimming at the same speeds. There were no mortalities and relative growth was similar among treatment groups. Average biopsy time for the notch removal method was lower and the wounds appeared to heal more quickly compared with the full removal method.     

Occurrence of the non-anadromous life history in the shirauo, Salangichthys microdon

Strontium (Sr) and calcium (Ca) concentrations in the otoliths of the shirauo, Salangichthys microdon, collected from Japanese coastal waters were examined by wavelength‐dispersive X‐ray spectrometry on an electron microprobe. The otolith Sr : Ca ratios gradually increased from 15.7?19.7 × 10^?3 around the core to 32.8?39.4 × 10^?3 around the edge of otoliths, with no transition point from the low Sr : Ca ratio phase to the high phase. The fluctuation pattern of otolith Sr : Ca ratios was different from the anadromous form reported in a previous study. The evidence indicates that the fish do not necessarily migrate into freshwater during the growth phase.     

Diel pattern of fish presence at the Changzhou fishway (Pearl River,China) during the flood season

Restoration of fish populations is required in China with the use of fishways at dammed rivers, but scant biological data is available for their design and operation. The Changzhou fishway (CFW) on the Pearl River is the largest in China, providing a channel for multispecies upstream migrations. This preliminary study used a combination of fish traps and hydroacoustic detection to document the diurnal presence of fish in the CFW, with the purpose of providing essential data for a future, solid performance of this fishway. Eighteen species from a total of 1362 fish individuals were collected in the traps. Three species were dominant, namely, Squaliobarbus curriculus, Pseudobagrus vachellii and Cirrhinus molitorella, altogether sharing proportions of 74.70 and 76.26% in the daytime and nighttime, respectively. Balanced by the up‐/downward movements of each fish, they entered the fishway with net numbers of 339.9 ± 113.4 ind.h^?1, and exited with net numbers of 234.54 ± 26.66 ind.h^?1. In diel fishway use, no significant differences were observed in either species composition or individual size during a diurnal period. Fish presence at the monitored sites within the fishway was recorded primarily during the daytime, showing a particular trend towards crepuscular use at the entrance and an uninterrupted use at the exit. This initial study showed a significant fish presence in the CFW during the flood season; the outcome of the study will be the basis of future research aimed at evaluating the passage efficiency of the CFW and the use thereof in other seasons.     

Immersion mass marking of pikeperch (<Emphasis Type="Italic">Sander lucioperca</Emphasis>) larvae in oxytetracycline hydrochloride and its detection using fluorescence microscopy

Fish marking is essential to distinguish between stocked and naturally reproduced fish. This procedure is challenging, especially in early life stages. Therefore the objective of this study was to investigate the possibility of oxytetracycline hydrochloride (800 mg L^?1 solution) immersion marking on 3–5 days old pikeperch larvae, with focus on durability and readability of marks on the otolith of marked individuals. The immediate mortality after marking was low (?5%) and growth of the pikeperch was continuous during the eight week study period. The fluorescent marks were detectable during the whole study period, but after the sixth week grinding of the otoliths was necessary. With grinding otoliths the processing time and the probability of damaging the mark increased significantly. Another critical period was the necessity of sample storage in a dark place due to the marks’ instability in light, in all phases before the final mark detection. The study confirmed the suitability of oxytetracycline hydrochloride marking for use in studies focused on pikeperch juveniles’ early growth, stocking success, distribution etc.     

Validation and efficacy of transgenerational mass marking of otoliths in viviparous fish larvae

Transgenerational mass marking of viviparous fish larvae in vivo was validated by intra‐muscular injection of elemental strontium chloride (SrCl₂) in gestating females and detection of the Sr in the otoliths of developing larvae. All otoliths of brown rockfish Sebastes auriculatus larvae produced from SrCl₂‐injected females showed enriched Sr:Ca ratios near the otolith edges, and the signatures did not appear to be affected by the anterior, centre and posterior positions of larvae within the ovary. Results from the present study indicate that transgenerational marking is a highly reliable technique for marking large numbers of extremely small viviparous fish larvae.     

Exploitation and mortality rates of European hake (Merluccius merluccius) in the Aegean Sea (Izmir Bay,Turkey)

Experimental trawl surveys in Izmir Bay (Aegean Sea) were taken seasonally between 2007 and 2009. A total of 1353 specimens were sampled ranging from 5.9 cm (1.4 g) to 44.4 cm (670 g). The main size group of Merluccius merluccius was between 14 and 25 cm total length (TL). In total, the mean catch per unit effort (CPUE) for hake by number and weight was 101.4 ± 18.4 ind h^?1 and 10.7 ± 1.5 kg h^?1, respectively. Highest mean CPUE by number and biomass was determined as 257.1 ± 15.1 ind h^?1 and 15.1 ± 4.9 kg h^?1 in summer. Mortality (M, F, Z) ratios of hake were 0.58 year^?1, 1.66 year^?1 and 2.24 year^?1, respectively. Because of the many smaller specimens caught in this study, this seems to indicate that there may be heavy fishing pressures on the hake population; the high exploitation rate (E = 0.74) appears to confirm this conclusion.     

Influences of salinity on the physiology and distribution of the Arctic coralline algae,Lithothamnion glaciale (Corallinales,Rhodophyta)

In Greenland, free‐living red coralline algae contribute to and dominate marine habitats along the coastline. Lithothamnion glaciale dominates coralline algae beds in many regions of the Arctic, but never in Godthåbsfjord, Greenland, where Clathromorphum sp. is dominant. To investigate environmental impacts on coralline algae distribution, calcification and primary productivity were measured in situ during summers of 2015 and 2016, and annual patterns of productivity in L. glaciale were monitored in laboratory‐based mesocosm experiments where temperature and salinity were manipulated to mimic high glacial melt. The results of field and cold‐room measurements indicate that both L. glaciale and Clathromorphum sp. had low calcification and photosynthetic rates during the Greenland summer (2015 and 2016), with maximum of 1.225 ± 0.17 or 0.002 ± 0.023 μmol CaCO ₃ · g^?1 · h^?1 and ?0.007 ±0.003 or ?0.004 ± 0.001 mg O₂ · L^?1 · h^?1 in each species respectively. Mesocosm experiments indicate L. glaciale is a seasonal responder; photosynthetic and calcification rates increase with annual light cycles. Furthermore, metabolic processes in L. glaciale were negatively influenced by low salinity; positive growth rates only occurred in marine treatments where individuals accumulated an average of 1.85 ± 1.73 mg · d^?1 of biomass through summer. These results indicate high freshwater input to the Godthåbsfjord region may drive the low abundance of L . glaciale , and could decrease species distribution as climate change increases freshwater input to the Arctic marine system via enhanced ice sheet runoff and glacier calving.     

Effects of desiccation and rewetting on the release and decomposition of dissolved organic carbon from benthic macroalgae

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An osmotic induction method for externally marking saltwater fishes,Stigmatopora argus and Stigmatopora nigra,with calcein

Calcein marking via osmotic induction and overnight immersion was compared in saltwater fishes. Immersion in a salt solution for 10 min followed by 30 min in a 500 mg l^?1 calcein solution produced the highest fluorescent mark in 40 min with no effect on survival.     

Anesthesia of wild female Persian sturgeon,Acipenser persicus (Borodin, 1897) breeders during controlled propagation: effects on hematological parameters,stress response and reproductive performance

This study examined the effects of anesthesia on the hematological and biochemical parameters as well as the reproductive performance of wild female Persian sturgeon, Acipenser persicus, during controlled spawning. Fourteen mature females were divided into two groups: ‘anesthetized’ and ‘non‐anesthetized’. All activities including transportation, catheterizing and handling were performed with both groups: (i) under anesthesia (150 ppm clove oil), and (ii) without anesthesia. After 10 days storage and handling, blood samples were taken from all fish after anesthesia. No significant differences were found in the reproductive performance of either group. However, differences were found in the hematological parameters, with values being significantly higher in the non‐anesthetized group, including neutrophils (34.36 ± 6.33% vs 23.63 ± 5.22%), monocytes (2.84 ± 1.70% vs 1.27 ± 0.64%), mean corpuscular volume (321.3 ± 39.40 pg vs 228.0 ± 24.46 pg) and mean corpuscular hemoglobin (106.9 ± 15.70 fl vs 76.50 ± 7.50 fl). Significantly lower values were found in the non‐anesthetized group for lymphocytes (60.68 ± 7.25% vs 73.54 ± 4.80%), Hb (4.62 ± 0.74 mg dl^?1 vs 6.28 ± 1.21 mg dl^?1), Hct (13.86 ± 1.76% vs 18.84 ± 3.85%), red blood cell (0.43 ± 0.05 cell mm^?3 vs 0.85 ± 0.13 × 10⁶ cell mm^?3) and white blood cell (22 403 ± 2240 cell mm^?3 vs 35 318 ± 3084 cell mm^?3). The non‐anesthetized fish had significantly higher cortisol levels compared to the anesthetized group (62.33 ± 8.85 ng ml^?1 vs 46.12 ± 8.07 ng ml^?1). There was no difference in plasma glucose levels between groups. It is concluded that the use of clove oil as an anesthetic is suitable for handling of wild female Persian sturgeon in controlled propagation programmes. However, further research is needed to determine a standardized protocol for the safe application of anesthesia for use in sturgeons in general.     

Bimodal oxygen uptake in a freshwater air-breathing fish,Notopterus chitala

Measurements of bimodal oxygen uptake have been made in a freshwater air-breathing fish,Notopterus chitala at 29.0±1(S.D.)°C. xhe mean oxygen uptake from continuously flowing water without any access to air, was found to be 3.58±0.37 (S.E.) ml O₂ · h^?1 and 56.84+4.29 (S.E.) ml O₂ · kg^?1 · h^?1 for a fish weighing 66.92 + 11.27 (S.E.) g body weight. In still water with access to air, the mean oxygen uptake through the gills were recorded to be 2.49 ± 0.31 (S.E.) ml O₂ · h^?1 and 38.78 ± 1.92 (S.E.) ml O₂ · kg^?1 · h^?1 and through the accessory respiratory organs (swim-bladder) 6.04±0.87 (S.E.) ml O₂ · h^?1 and 92.32±2.91 (S.E.) ml O₂ · kg^?1 · h^?1 for a fish averaging 66.92±11.27 (S.E.) g. Out of the total oxygen uptake (131.10 ml O₂ · kg^?1 · h^?1), about 70% was obtained through the aerial route and the remainder 30% through the gills.     

Effects of clove oil,essential oil of Lippia alba and 2‐phe anaesthesia on juvenile meagre,Argyrosomus regius (Asso, 1801)

The objectives of this experiment were to (i) determine the efficacy of essential oils of clove (CO) and Lippia alba (EOLA) to induce deep anaesthesia in juvenile specimens (49.0 ± 6.2 g body mass, 16.6 ± 0.8 cm; n = 8 per treatment) of meagre (Argyrosomus regius); and (ii) study the feasibility of these substances, together with 2‐phenoxyethanol (2‐PHE), as potential sedatives [low concentration: (i) EOLA: 12 mg L^?1; (ii) CO: 1 mg L^?1; and (iii) 2‐PHE: 33 mg·L ^?1; n = 8 per treatment] for live fish transport of this species. All test were performed at a constant temperature (18°C). Thus, the main primary stress indicator (plasma cortisol) and secondary factors (plasma metabolites) were evaluated. In addition, growth hormone (GH) mRNA expression was also evaluated in the pituitary gland. The results indicated that EOLA is considered to be effective for deep anaesthesia when the concentration is close to 160 mg L^?1, while CO produces the same effect when lower concentrations are added (40–50 mg L^?1). Regarding sedative concentrations, a significant ~3‐fold increase in plasma cortisol levels was detected in the EOLA group when compared to control specimens. In addition, glucose levels were not reduced and significantly increased (~1.6‐fold) for 2‐PHE in relation to the control fish. None of the anaesthetics promoted a significant difference for GH expression with respect to the control group, but a significant ~2‐fold increase for 2‐PHE treatment with respect to the EOLA exposition was found in this gene expression. Results show that none of the anaesthetics analysed, at least in the ranges of concentrations used in this study (EOLA 12 mg L^?1, CO 1 mg L^?1, 2‐PHE 33 mg L^?1), are recommended for live fish transport, as shown by the absence of inhibition on the stress parameters assessed.