Improved performance of gilthead sea bream, Sparus aurata, larvae after ozone disinfection of the eggs

Ozone (O₃) dissolved in seawater (ODS) was evaluated, as an egg disinfectant, on the spawn of captive gilthead sea bream, Sparus aurata, brood stock. Four contact times (CT) were tested (0.6, 1.2, 2.4 and 4.8 mg min L⁻¹) where CT was calculated by multiplying the dissolved O₃ concentration (0.3 mg L⁻¹) by different exposure periods (2, 4, 8, 16 min). There was also a disinfected seawater treatment that contained no O₃ or derived compounds (CT 0) and an untreated seawater control. All ODS treatments reduced egg surface bacterial counts to zero, which was significantly (P<0.05) lower than the CT 0 and the control groups (194 and 1320 plate⁻¹ respectively). Nevertheless, the hatching rate was high in the control and the CT treatments 0, 0.6 and 1.2 (88.7%, 87.3%, 89.5% and 83.7% respectively) while eggs exposed to a CT 2.4 and 4.8 hatched poorly (36.5% and 20.4% respectively), which was likely due, at least in part, to larvae unable to break the egg chorion successfully. Swim‐bladder inflation was significantly higher in the ODS groups (>97%) compared with the control and CT 0 treatments (ca. 70%). The results suggest that a 2‐min exposure of eggs to 0.3 mg O₃ L⁻¹ of ODS (CT 0.6) would improve current protocols in marine larviculture.     

Chemical surface disinfection of eggs of Baltic cod,Gadus morhua L.

The effect of two disinfectants on eggs and larvae of Baltic cod, Gadus morhua, was investigated. The eggs were disinfected for 10 min using various concentrations of either glutaraldehyde (100, 200, 400, 600 and 800 mg L^?1) or iodophor (10, 50, 100 and 150 mg L^?1), 1–4‐days post‐fertilization. Bactericidal effect of disinfection, survival to hatching, hatching success and larval abnormalities were assessed. Larval survival was recorded at 5‐, 10‐ and 15‐days post‐hatch (dph). Although Baltic cod eggs have an unusually thin chorion, they could tolerate surface disinfection. A reduction in bacterial growth was observed with increased concentrations of disinfectant (3.0 × 10⁷–1.6 × 10¹ CFU mL^?1). Abnormalities in newly hatched larvae were not related to disinfection. Survival of the yolk sac larvae was significantly better for eggs treated with 400 mg L^?1 glutaraldehyde for 10 min at 10 and 15 dph. Effective disinfection was also recorded using 100 mg L^?1 Actomar K30. Egg batch effect rather than initial bacterial concentration, disinfectant type or incubation method determined the survival of the eggs to hatching and survival of larvae. Because of the carcinogenic effect of glutaraldehyde, iodophor is recommended for routine disinfection of cod eggs.     

Hatch rate of channel catfish Ictalurus punctatus (Rafinesque 1818) eggs treated with 100 mg L−1 copper sulphate pentahydrate

Catfish hatcheries use copper sulphate pentahydrate (CuSO₄·5H₂O) as an economical control for saprolegniasis on eggs. This study determines hatch rate of channel catfish, Ictalurus punctatus (Rafinesque 1818), eggs in hatching troughs containing 23.8 °C flow‐through well water when treated with 100 mg L^?1 CuSO₄·5H₂O (10 times the proposed therapeutic dose). Eggs were treated daily until the embryos reached the eyed stage. Fry survival in the control and 100 mg L^?1 CuSO₄·5H₂O treatments was significantly different (15% and 71% respectively). This study demonstrates that there is a considerable margin of safety in using CuSO₄·5H₂O as a catfish egg treatment to control saprolegniasis.     

Effects of 2-phenoxyethanol on survival of normal juveniles and malformed juveniles having lordosis or nonfunctional swimbladders of European sea bass (Dicentrarchus labrax L., 1758)

The objectives of this study were to evaluate the effects of 2‐phenoxyethanol (2‐PE), which is an anaesthetic, on survival rates of normal juveniles and malformed juveniles having lordosis or nonfunctional swim bladders of European sea bass (Dicentrarchus labrax L., 1758) and to establish the LC₅₀ (the concentration lethal to 50% of test animals at concentrations of 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4 and 4.5 mL L⁻¹) and LT₅₀ (the time lethal to 50% of test animals after 10‐, 20‐, 30‐, 40‐, 50‐ and 60‐min time periods) of 2‐PE at 19±0.5°C, salinity 38 g L⁻¹, pH 7.4–7.8 and dissolved oxygen >8 mg L⁻¹. Between concentrations of 0.05 and 0.25 mL L⁻¹, 2‐PE did not cause any mortality or toxicity on normal, lordosis and nonfunctional swimbladder juveniles of sea bass during the 60‐min exposure period. On the other hand, significance in each group fish in their mortality rates between concentrations of 0.30 and 0.45 mL L⁻¹ was observed (P<0.05). The nonfunctional swimbladder juveniles showed lower LC₅₀ than normal and lordosis juveniles respectively. Also, nonfunctional swimbladders juveniles showed lower LT₅₀ than normal and lordosis juveniles respectively. At concentrations of 0.30, 0.35, 0.40 and 0.45 mL L⁻¹, induction times were found to be significantly different among the three groups (P<0.05). Recovery times were not found to be significantly different in two groups at concentrations of 0.30 and 0.40 mL L⁻¹ (P>0.05). The toxic effect of 2‐PE on sea bass juveniles increased depending on the exposure times (P<0.05). The most suitable concentrations of 2‐PE were 0.30–0.35 mL L⁻¹ between minutes 10 and 30, although the normal juveniles can resist to 0.45 mL L⁻¹ of 2‐PE concentration for 20 min. The 2‐PE showed toxicity in relation to the concentrations and exposure time combinations among the three groups in the order; nonfunctional swimbladder fish >lordosis fish >normal fish.     

The use of tannic acid to remove adhesiveness from pikeperch, Sander lucioperca, eggs

The studies conducted in 2003–2004 focused on the possibilities of applying a tannic solution to remove adhesiveness from pikeperch eggs. Spawners were caught in Lake P?tnowskie (central Poland) and then transported to the Gos?awice Fish Farm. After initial selection, the fish were weighed, measured and stimulated with human chorionic gonadotropin. Gametes were obtained 5 days after the first injection. The weight and diameter of the eggs, and the commercial fecundity of individual females were determined. The eggs were fertilized with the dry method. After the addition of water, the eggs were mixed for 4 min, and then divided into 20 g portions. After determining the number of eggs in the various portions, the adhesiveness removal procedure was performed. Three concentrations of tannic acid solution (500, 1000, 1500 mg L^?1) and three exposure times (0.5, 2, 5 min) were applied. The eggs were incubated in Weiss jars. The studies indicated that both the solution concentration and the exposure time significantly (P<0.05) impacted pikeperch egg hardening, the degree of adhesive removal and embryo survival. The tannic acid solution concentration of ≤500 mg L^?1 applied for 0.5–2 min was not effective; the eggs clumped and it was impossible to separate them even with intensive mixing. Better results were obtained using higher tannic acid concentrations and/or by lengthening exposure time. The adhesiveness of pikeperch eggs disappeared completely after 5 min exposure to tannic acid solution concentrations of 500–1500 mg L^?1 or after 2 min exposure to solution concentrations of 1000–1500 mg L^?1. In these variants, the embryo survival rate to the eyed‐egg stage was 78.0–84.0% (2003) and 82.3–84.7% (2004). However, high tannic acid concentration had a negative impact on the pikeperch larvae hatching. The greatest decrease in survival rate was observed in groups exposed to a tannic acid solution of 1500 mg L^?1 for 2 and 5 min periods. Thus, the optimum method for removing pikeperch egg adhesiveness was to apply a solution of tannic acid at a concentration of 500 mg L^?1 for 5 min or 1000 mg L^?1 for 2–5 min.     

Hydrogen peroxide oxygenation and disinfection capacity in recirculating aquaculture systems

Hydrogen peroxide (H₂O₂) treatment is an alternative for disinfection in aquaculture, which may be advantageous as it dissociates and disinfects while increasing water oxygen concentration. Yet, accurate dosing remains undeveloped in Recirculating Aquaculture Systems (RAS). Dosage requirements can depend on organic burden, stocking density, feeding frequency, salinity, temperature and biofilter performance. The present case study investigated the dual effect of H₂O₂ application for oxygen enrichment and disinfection when continuously applied to a RAS rearing European seabass. H₂O₂ addition equivalent to 2.4 and 15.8 H₂O₂ mg L⁻¹ were applied for 4 h per day in three 5-days experiments. H₂O₂ was injected at the inlet of protein skimmer and/or the rearing tanks in or without combination with traditional disinfection methods. Water microbial load and oxygen saturation were determined, along with stress markers glucose and cortisol in blood plasma of fish. Doses of 15.8 mg L⁻¹ H₂O₂ steadily increased oxygen levels in holding tank water from ∼50 % to over 100 % saturation while reducing microbial load (from 604.4 CFU ml⁻¹ in the rearing tanks before dosing to 159.8 CFU ml⁻¹ after application), achieving suitable conditions for commercial fish densities in RAS. The doses used had negligible impact on biofilter performance and did not affect the fish in terms of stress. Overall results indicate H₂O₂ is effective for disinfection and oxygenation of RAS systems when applied at appropriate dosage and we recommend the protein skimmer as the safest position in order to protect the bacterial community of the biofilters and the reared fish.     

Effect of ammonia-N on growth and feeding of juvenile Macrobrachium rosenbergii (De-Man)

Survival rate, growth and feed intake were determined for late juveniles (4.31 ± 0.18 g) of river prawn Macrobrachium rosenbergii in freshwater with total ammonia‐N (NH₃‐N+NH₄‐N) concentrations of 0.015 (control), 0.5, 1.0 and 1.5 mg L⁻¹ for 60 days at pH 7.53 ± 0.04 and temperature 24.0 ± 2.5°C. Survival rate was significantly (P<0.05) lower (54 ± 4.2–70 ± 5.4%) for total ammonia concentrations from 0.5 to 1.5 mg L⁻¹ [0.0139–0.0419 mg L⁻¹ of unionized ammonia (NH₃)]. Growth (0.026–0.030 g day⁻¹ range) of the prawns did not differ for the different NH₃ levels but were significantly (P<0.05) lower compared with control (0.056 g day⁻¹). Feed intake rates also diminished significantly (P<0.05) from 77.60 ± 2.45% at control (0.015 mg L⁻¹ NH₃‐N) to 48.69 ± 2.13% at 1.5 mg L⁻¹ NH₃‐N (0.0419 mg L⁻¹ of unionized NH₃).     

Preliminary success using hydrogen peroxide to treat Atlantic salmon,Salmo salar L., affected with experimentally induced amoebic gill disease (AGD)

Currently, the only effective and commercially used treatment for amoebic gill disease (AGD) in farmed Tasmanian Atlantic salmon is freshwater bathing. Hydrogen peroxide (H₂O₂), commonly used throughout the aquaculture industry for a range of topical skin and gill infections, was trialled in vitro and in vivo to ascertain its potential as an alternative treatment against AGD. Under in vitro conditions, trophozoites of Neoparamoeba perurans were exposed to three concentrations of H₂O₂ in sea water (500, 1000 and 1500 mg L^?1) over four durations (10, 20, 30 and 60 min) each at two temperatures (12 and 18 °C). Trophozoite viability was assessed immediately post‐exposure and after 24 h. A concentration/duration combination of 1000 mg L^?1 for >10 min demonstrated potent amoebicidal activity. Subsequently, Atlantic salmon mildly affected with experimentally induced AGD were treated with H₂O₂ at 12 and 18 °C for 15 min at 1250 mg L^?1 and their re‐infection rate was compared to freshwater‐treated fish over 21 days. Significant differences in the percentage of filaments affected with hyperplastic lesions (in association with amoebae) and plasma osmolality were noted between treatment groups immediately post‐bath. However, the results were largely equivocal in terms of disease resolution over a 3‐week period following treatment. These data suggest that H₂O₂ treatment in sea water successfully ameliorated a clinically light case of AGD under laboratory conditions.     

Acute toxicity of nitrite to mud crab Scylla serrata (Forsskål) larvae

Early larval stages of mud crab Scylla serrata were exposed to different concentrations of nitrite (40, 80 and 160 mg L⁻¹ and a control, without added nitrite) and three salinity levels (25, 30 and 35 g L⁻¹) using a static renewal method. No interactive effect of nitrite and salinity was detected. Estimated LT₅₀ in 96‐h toxicity tests decreased in all stages with increasing nitrite concentrations in all salinity levels. The 96‐h LC₅₀ values of nitrite‐N were 41.58, 63.04, 25.54, 29.98 and 69.93 mg L⁻¹ for zoea 1, 2, 3, 4 and 5 respectively. As the larvae grew, they showed a progressive increase in tolerance to nitrite. The toxicity of nitrite to larvae increased with exposure time. The median lethal concentration was not affected by salinity. The chloride component of salinity within 25–35 g L⁻¹ did not seem to be as effective in alleviating toxicity as has been reported in other crustacean species. Based on 96‐h LC₅₀ and an application factor of 0.1, the ‘safe level’ of rearing mud crab larvae was calculated to be 4.16, 6.30, 2.55, 2.99 and 6.99 mg L⁻¹ nitrite‐N for zoea 1, 2, 3, 4 and 5 respectively.     

Tolerance of Penaeus monodon Fabricius embryos to ozonated seawater

The tolerance of Penaeus monodon embryos from five spawnings (families) to four different ozone doses in seawater [0.0, 0.5, 1.0 and 2.0 mg L⁻¹, measured by the residual oxidant concentration (ROC)] was examined when applied for three exposure times (1, 2 and 4 min) at three post‐spawning treatment times (25, 120 and 480 min post‐spawning). Ozone dose typically had a larger affect on embryo hatching than exposure time and the ozone dose × exposure time interaction for most combinations of family and post‐spawning treatment time. At ozone doses of 2.0 mg L⁻¹, embryos had lower hatchings than controls for all families at 25 and 120 min post‐spawning, and for several combinations of family and exposure time at 480 min post‐spawning. At ozone doses of 1.0 mg L⁻¹, the effect on embryo hatching was more varied between families and exposure times for the three post‐spawning treatment times, but typically embryos were less affected when exposed at later post‐spawning treatment times. Ozone doses of 0.5 mg L⁻¹ typically had minimal effects on hatching for all exposure times and post‐spawning treatment times. In summary, later‐stage P. monodon embryos typically tolerated ozone doses of up to 1.0 mg L⁻¹ in seawater for durations of up to 4 min.     

Efficacy of formalin and povidone–iodine disinfection techniques on the eggs of three marine finfish species

Surface disinfection trials were performed on eggs from three marine finfish species: California yellowtail (CYT; Seriola lalandi), white seabass (WSB; Atractoscion nobilis) and California halibut (HA; Paralichthys californicus). All three species were spawned from captive populations maintained at the Hubbs‐SeaWorld Research Institute (HSWRI). Five disinfection treatments were used for each species; Treatment 1 included 100 mg L^?1 of formalin (F100) for 60 min (current HSWRI treatment), Treatment 2 included 1000 mg L^?1 of formalin for 15 min (F1000), Treatment 3 included povidone–iodine of 50 mg L^?1 for 15 min (PI50), Treatment 4 included povidone‐iodine of 100 mg L^?1 for 10 min (PI100) and Treatment 5 involved a control with no chemical treatment (CONT). For each treatment, the per cent egg hatching rate, per cent survival to first feeding and notochord length at the time of hatching to the nearest 0.1 mm were recorded. Bacteria were also cultured from eggs after treatment to determine the effectiveness of each treatment in reducing the bacterial counts (CFU mL^?1). Treatments F100, F1000 and CONT yielded the highest hatch rates for each species (70–80%), whereas treatments PI50 and PI100 yielded the lowest hatch rates (0–2%). There were no significant differences in survival to first feeding or notochord length, which suggests that the disinfection treatments did not have a negative effect on the yolk sac larvae. The PI50 and PI100 treatments had the lowest bacterial colony counts, showing almost zero bacterial growth. The highest bacterial growth occurred in the F100, F1000 and CONT treatments. Based on the results from this study, the F100 treatment provided the best balance of disinfection and larval health for CYT, WSB and HA.     

Comparison of iodine and glutaraldehyde as surface disinfectants for red porgy (Pagrus pagrus) and white sea bream (Diplodus sargus sargus) eggs

The efficacy of iodine and glutaraldehyde as fish egg surface disinfectants were assessed in red porgy (Pagrus pagrus) and white sea bream (Diplodus sargus sargus) eggs, two species of interest for Mediterranean aquaculture. Iodine was effective in reducing the bacterial load of the 1-day-old eggs when applied at 50 mg L⁻¹ for 5 min. The same concentration did not cause any significant change in hatching success or survival of the larvae for the first 5 days. Glutaraldehyde failed to reduce the bacterial load of the fish eggs at concentrations that were safe for the eggs (100 mg L⁻¹ for 5 min), as it had a significant effect in preventing hatching of the developed embryo. Disinfecting 0-day-old eggs with iodine resulted in a significant reduction of hatching percentage, while larval survival thereafter was unaffected. The results of the present study suggest that iodine may be an appropriate egg disinfectant for both red porgy and white sea bream.     

Efficacy of formalin,iodine and sodium chloride in improvement of egg hatching rate and fry survival prior to the onset of exogenous feeding in yellow perch

Egg disinfection is considered the most important routine work in hatcheries to avoid fungal and/or bacterial infection of fish eggs. The aim of this study was to determine the effectiveness of three disinfectants: formalin, iodine and sodium chloride on the hatching success of yellow perch eggs. The disinfectants were tested in triplicate at different concentrations for 15 and 30 min bath treatments. Two experiments were conducted; formalin at five concentrations (25, 50, 100, 150 and 200 mg L^?1) and 25 mg L^?1 iodine were tested in the first experiment. The second experiment involved formalin at three concentrations (250, 500 and 1000 mg L^?1), iodine at three concentrations (50, 100 and 250 mg L^?1) and sodium chloride at three concentrations (500, 1000 and 3000 mg L^?1) were used. Iodine and sodium chloride‐treated eggs hatched earlier than formalin‐treated eggs. The highest mean percentage of eyed stage, hatching rate and survival to first feeding fry was observed at 200 mg L^?1 formalin for 30 min, 50 mg L^?1 iodine for 15 min and 500 mg L^?1 sodium chloride for 30 min. High concentrations of formalin (1000 mg L^?1), iodine (250 mg L^?1) and sodium chloride (1000 and 3000 mg L^?1) showed toxicity to yellow perch eggs, resulting in low hatching rate and survival to first feeding fry. We recommended formalin at a concentration of 150–200 mg L^?1 for 30 min as an effective, easily available and low‐cost disinfectant for routine use to improve yellow perch hatchability.     

The use of clove oil, metomidate, tricaine methanesulphonate and 2-phenoxyethanol for inducing anaesthesia and their effect on the cortisol stress response in black sea bass (Centropristis striata L.)

Juvenile and adult black sea bass (Centropristis striata L.) were exposed to various concentrations of four anaesthetics to determine practical dosages for handling as well as for procedures such as bleeding, ovarian biopsy or tag implantation. In experiment 1, juveniles exposed to either 2.0 mg L^?1 metomidate, 15 mg L^?1 clove oil, 70 mg L^?1 tricaine methanesulphonate (TMS) or 200 mg L^?1 2‐phenoxyethanol (2‐PE) reached stage II of anaesthesia in 3–5 min and could be handled for weighing and measuring. All fish had completed recovery to stage III within 6 min. In experiment 2, the established concentrations of each anaesthetic were tested on juveniles to determine their ability to prevent a reflex to a subcutaneous needle puncture. All of the fish exposed to clove oil (20 mg L^?1) and 40% of the TMS‐treated (70 mg L^?1) fish reacted while none of the fish anaesthetized in metomidate (2.0 mg L^?1) or 2‐PE (200 mg L^?1) responded to the needle puncture. In experiment 3, metomidate (5.0 mg L^?1), clove oil (30 mg L^?1) TMS (125 mg L^?1) or 2‐PE (300 mg L^?1) were all effective for performing an ovarian biopsy or tag implantation on adults. In experiment 4, TMS (125 mg L^?1) exacerbated the cortisol response to a short handling stressor during a 30 min exposure. Fish anaesthetized in 2‐PE (300 mg L^?1), metomidate (5.0 mg L^?1) or clove oil (40 mg L^?1) had increased cortisol levels associated with the handling stressor but there were no further increases during the remainder of the experimental period. The results demonstrate that these anaesthetics are effective for sedation and anaesthesia of black sea bass and that the best choice is dependant upon the procedures to be performed.     

Growth, stress response and free amino acid levels in Senegalese sole (Solea senegalensis Kaup 1858) chronically exposed to exogenous ammonia

Stressful husbandry conditions are likely to affect growth and amino acid metabolism in fish. In this study, chronic ammonia exposure was used to test the effects of a stressor on growth and amino acid metabolism of Senegalese sole juveniles. The fish were exposed for 52 days to 11.6 mg L⁻¹ [low‐TAN (L‐TAN)] or 23.2 mg L⁻¹ [high‐TAN (H‐TAN)] of total ammonia nitrogen (TAN), or to 0 mg L⁻¹ (Control). Growth in L‐TAN groups was slightly but significantly different from the Control groups [relative growth rate (RGR=0.35±0.13 and 0.52±0.23% day⁻¹ respectively)]. In H‐TAN groups, growth was severely affected (RGR=0.01±0.13% day⁻¹). Stress parameters (plasma cortisol and glucose) showed slight or no significant differences between treatments. Plasma free amino acid (FAA) concentrations were affected after H‐TAN treatment. Increases in glutamine and aspartate concentrations in H‐TAN fish suggest alterations in amino acid metabolism related to nitrogen excretion processes. Some of the changes in FAA concentrations also suggest mobilization to energy supply and synthesis of metabolites related to stress response. Therefore, Senegalese sole seem to adapt to the L‐TAN concentration tested, but the H‐TAN concentration reduced growth and affected amino acid metabolism.     

Effect of the coupled action of ultrasonic waves and advanced oxidation reaction on the properties of bottom sediments originating from trout culture

The objective of this study was to determine the effect of preliminary ultrasound treatment of bottom sediments, a factor enhancing the reaction of advanced oxidation, on changes in the properties of sediments originating from the intensive aquaculture of rainbow trout. Experiments were conducted on a laboratory scale in model reactors with an active volume of 1.0 L, either under conditions of exposure to the ultrasonic field alone or under conditions of exposure to both the ultrasonic field and a chemical reagent: Fe³⁺/H₂O₂. The results we achieved showed that preliminary ultrasound treatment of bottom sediments increased the advanced oxidation reaction yield and consequently improved the physicochemical properties of the sediments. The highest yield was obtained upon 15‐min ultrasound treatment at a Fenton reagent dosage of 1.5 g Fe³⁺ L^?1 and g H₂O₂ L^?1. These conditions enabled a 43% reduction in the concentrations of organic compounds in the filtrate, shortened the capillary suction time (CST) by 93%, decreased the concentration of volatile substances in the sediments by 13% and decreased the levels of total nitrogen in sediments by 9% and of total phosphorus in the filtrate by 21%, and additionally caused a decrease in the mineral and dry matter content in the sediments.     

Reduction of in vitro growth in Flavobacterium columnare and Saprolegnia parasitica by products containing peracetic acid

Commercial products containing peracetic acid (PAA) are strong disinfectants with a wide spectrum of antimicrobial activity and have been suggested as potential therapeutic agents in aquaculture. The aim of this study was to compare the in vitro reduction of growth on two fish pathogens, Flavobacterium columnare and Saprolegnia parasitica, by seven commercial PAA‐containing products. Flavobacterium columnare was exposed to 1, 2, 4, 6, 8 and 10 mg L^?1 PAA and S. parasitica was exposed to 0.5, 1, 2, 4, 6, 8 and 10 mg L^?1 PAA in petri dishes for 24 h incubation. The reduction of growth was measured in comparison to a PAA‐free control. A reduction of the growth was observed for both pathogens with increasing PAA concentration. Hydrogen peroxide (H₂O₂) possibly has a role in the effectiveness of the products, since products with lower PAA concentrations had a higher concentration of H₂O₂. The commercial products with a low concentration of PAA and a low PAA:H₂O₂‐ratio were generally more effective against pathogens. The practical application of the products with low PAA concentration should be prioritized.     

Clove oil as an anaesthetic for common octopus (Octopus minor, Sasaki)

We evaluated the anaesthetic effect of clove oil [2‐methoxy‐4‐2‐(2‐propenyl)‐phenol] on the common octopus (Octopus minor), in terms of the time required to become anaesthetized (‘anaesthetic time’) and recovery time. We used a factorial experimental design and administered clove oil at different temperatures (15, 20 and 25°C) and concentrations (50, 100, 150, 200, 250 and 300 mg L⁻¹). We observed a significant (P<0.05) relationship between concentration and temperature, and each variable was effective (P<0.05). Anaesthetic time linearly decreased as the concentration and temperature increased. However, recovery time increased as the concentration increased and temperature decreased. There was no mortality. A concentration of 200 mg L⁻¹ clove oil showed rapid anaesthetic and recovery times in the common octopus, indicating its suitability for this species.     

Effects of four egg desticking procedures on hatching rate and further survival and growth of larvae in the tench (Tinca tinca L.)

Four desticking procedures for tench eggs (A: tannic acid solution (1 g L⁻¹) for 15 s; B: alcalase enzyme solution (8 mL L⁻¹) for 60 s; C: alcalase enzyme solution (15 mL L⁻¹) for 120 s; D: Woynarovich and Woynarovich (1980) solution for 58 min followed by tannic acid solution (1 g L⁻¹) for 15 s) were tested to obtain data about influence on embryo survival to hatching stage and further survival and growth of the larvae. In the tannic acid and Woynarovich and Woynarovich (1980) treatment (A and D) few eggs stuck together and some were adhered to the incubator walls, whereas in the alcalase treatments (B and C) eggs neither stuck together nor adhered to the incubator walls. Percentages of hatched larvae did not show significant differences (mean values ranged between 47.4% in treatment A to 37.0% in treatment C). Larvae deformities observed were <0.5% in all cases. There were no significant differences among survival and growth rates of the larvae from different egg desticking origin, reaching, after 30 days, mean survival values around 90% and total length and weight of 12.5 mm and 19 mg respectively.