Effects of commonly used disinfectants on bacterial load,hatchability and survival of Bluefin Sea bream (Sparidentex hasta) eggs

The efficacy of four chemical reagents, iodophor, formalin, hydrogen peroxide and bronopol as fish egg surface disinfectants were evaluated in bluefin sea bream (Sparidentex hasta). Fertilized eggs were counted and subjected to a static bath dip treatment in different concentrations of the above chemicals for 4 min before being incubated at 20 ± 0.5°C for 40 h. Treatment efficacy of the different disinfectants was evaluated by assessing the bactericidal activity, egg hatch percentage and survival of larvae up to 3 days post hatch. Results showed that iodophor at medium concentrations (75 and 100 ppm) was the best of all tested disinfectants in bacterial killing ability (12% reduction in the bacterial counts), egg hatching per cent (99.8% and 99.6% respectively) and larval survival up to 3 days post hatch (50.8% and 54.8% respectively). Formalin was the second best disinfectant at levels of 100 and 150 ppm. Hydrogen peroxide gave good results compared with the control while, bronopol showed discouraging results. In conclusion, iodophor appeared to be suitable for bluefin sea bream eggs disinfection with a 4 min exposure to 75–100 ppm when applied 14–16 h after egg fertilization.     

Antifungal effect of Origanum onites essential oil as an alternative to formalin in the artificial incubation of narrow‐clawed crayfish (Astacus leptodactylus Eschscholtz, 1823)

As alternative to formalin, the antifungal effect of a plant product [Origanum onites L. (Lamiaceae) oil] was investigated for use in the artificial incubation of narrow‐clawed crayfish (Astacus leptodactylus Eschscholtz) eggs. For this purpose, this study was conducted as two experiments. In experiment I, the eggs were artificially incubated for 40 days. In experiment II, juveniles were cultured to determine effects of O. onites oil on juveniles for 30 days. The experimental groups were as follows: formalin (3500 ppm for 15 min), O. onites oil (300 ppm for 15 min, 700 ppm for 2 min and 1000 ppm as a dip treatment 15 split‐second) and a control (no treatment). In the experiment I, the highest hatching rate (86%) and survival rate of stage II juveniles (80%) were observed in 1000 ppm dip group. These results were similar to that of formalin group (85% and 79%) respectively. The control group exhibited the lowest hatching rate (49%) and stage II rate (42%) compared with the 1000 ppm dip group and 3500 ppm formalin treatments. However, other concentrations (300 and 700 ppm) of O. onites showed toxic effects on the eggs and there was no hatching. In the experiment II, the survival rate and growth performance of the crayfish juveniles were similar in all groups. This study indicated that the 1000 ppm O. onites dip treatment could be a good alternative to formalin for improved egg hatchability in the artificial incubation of crayfish eggs.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.     

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.     

The application of tannic acid to the elimination of egg stickiness at varied moments of the egg swelling process in pikeperch,Sander lucioperca (L.)

The aim of this article was to analyse the process of pikeperch, Sander lucioperca, egg swelling and to apply tannic acid to eliminate egg stickiness at different moments of the swelling process on artificially obtained eggs. The first experiment involved observation of egg swelling process and the second determined the effect of temperature (12, 14 and 16°C) on the egg swelling rate. The third experiment involved elimination of egg stickiness in a tannin solution (0.75 g L^?1) where eggs were submerged in a solution for 0.5, 1, 2 and 5 min – 5, 10, 15, 20, 25 and 30 min following gamete activation. The results indicate that the pikeperch egg swelling process lasts 30 min. It was found that the temperature did not affect the process duration. The results of the third experiment showed that the effectiveness of tannic acid application in egg stickiness elimination increases with time. The best result was obtained in groups of eggs submerged for 1 and 2 min (86.5% and 80.5% of larvae were obtained respectively) 30 min following the gamete activation. The results presented in this study for the first time indicate the possibility of highly effective procedure of egg stickiness elimination with tannic acid in pikeperch aquaculture.     

Effect of Salinity on Embryonic Development,Hatchability, and Growth of African Catfish,Clarias gariepinus,Eggs and Larvae

Abstract

Effects of salinity on embryonic development and growth of African catfish, Clarias gariepinus, eggs and larvae were studied. Eggs were incubated at 27-29°C in 2,4,6,8, and 10 ppt sodium chloride. Rate of embryonic development was delayed in all salt solutions by 15, 15,28 and 30 minutes, in 2,4,6, and 8 ppt sodium chloride, respectively, when compared with the control group (0% salt); total mortality occurred at 12 hours after gastrula stage in the 10 ppt concentration. Percentage hatching was 45.1,47.7, 59.5,49.2, and 26.6% while percentage deformity was 10.4, 16.1, 52.0, 28.6, and 71.6% in 0, 2, 4, 6, 8, and 10 ppt salt treatments, respectively. There were significant differences (P <0.05) in the hatching percentage and in deformity percentage between 4, 6, and 8 ppt. Rate of yolk absorption was significantly faster in the control and 2 ppt salt treatments, but slower in 4, 6, and 8 ppt. Rate of increase in length was slower with increasing salinity. The optimum salinity for African catfish eggs and was between 0-2 ppt and acceptable up to 6 ppt. The results suggest that increasing salinity delayed hatching and development of African catfish eggs and larvae, respectively, as well as increased the deformity of the larvae.     

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.     

Effect of dietary <Emphasis Type="SmallCaps">l</Emphasis>-tryptophan on cannibalism,survival and growth in pikeperch <Emphasis Type="Italic">Sander lucioperca</Emphasis> (L.) post-larvae

The effect of supplemented commercial diets with crystalline l-tryptophan (TRP—5, 10 and 20 g TRP kg^?1) on cannibalism, survival and growth parameters of pikeperch post-larvae (Sander lucioperca) was evaluated. Fifteen-day-old pikeperch larvae (mean weight—6.8 mg) were reared during the next 28 days (20.5 °C, 16L:8D) in glass aquaria in a recirculating aquaculture system. The enzyme-linked immunosorbent assay showed that TRP-supplemented diets were effective in increasing the levels of serotonin (5-HT) in the body tissue of pikeperch. TRP supplementation resulted in a slight decrease in both types of cannibalism, although the reduction in cannibalism impact did not amount to more than a few percent. TRP treatment had no significant influence on the final survival of pikeperch post-larvae (ranged from 20.1 ± 12.4 to 29.0 ± 12.9 %). However, contrary to the earlier studies conducted on other fish species, no significant difference in the growth rates and feeding behavior of pikeperch between TRP-fed and control group were found. The final body weight and growth rate ranged from 0.211 ± 0.014 to 0.243 ± 0.016 and from 12.19 ± 0.38 to 12.76 ± 0.35 % day^?1, respectively. To our knowledge, this is the first study reporting the effects of TRP supplementation on the cannibalism–survival–growth relations in fish in the post-larval stage.     

Effects of different bronopol treatments on final survival rates in the artificial incubation of crayfish eggs (Pacifastacus leniusculus,Astacidae)

Two experiments were conducted on the effects of bronopol in the artificial incubation of crayfish eggs (Pacifastacus leniusculus) with the aim to search an alternative to formaldehyde. In the first experiment, 50, 250, 500 and 1000 ppm bronopol and 3000 ppm formaldehyde (control) in periodical administrations were tested on a density of 6.6 eggs cm^?2. After 44 days of incubation, the highest survival was obtained with 1000 ppm bronopol (81.9% to stage 2 juvenile, with no significant difference from formaldehyde), whereas lower bronopol concentrations resulted in significantly lower survival. In the second experiment, 1000, 3000 and 5000 ppm bronopol and 3000 ppm formaldehyde (control) administered for 15 min every second day were tested on eggs at a density of 20 eggs cm^?2. After 78 days of incubation, bronopol at 3000 ppm allowed for a stage 2 juvenile survival rate of 65.0% (with no significant difference from formaldehyde), whereas significantly lower survival was obtained with 1000 ppm or 5000 ppm. This study shows that bronopol may constitute an alternative to formaldehyde in the artificial incubation of crayfish eggs. A concentration of 3000 ppm administered for 15 min every second day may be adequate even on long incubations at high densities (at least 20 eggs cm^?2, one complete layer).     

A method for collecting eggs of Pseudocapillaria tomentosa (Nematoda: Capillariidae) from zebrafish Danio rerio and efficacy of heat and chlorine for killing the nematode's eggs

Pseudocapillaria tomentosa is a common pathogen of zebrafish (Danio rerio) in research facilities. We developed a method to collect and concentrate the nematode eggs using a modified sugar centrifugation method and documented their normal development. Embryonating stages with blastomere formation followed by elongation of the embryo prior to larva formation cumulated in developed larvae inside the eggs and hatching after 5–10 day. We then evaluated the efficacy of heat and chlorine to kill them based on a larva development assay. Eggs were exposed to 40, 50, 60 °C for 30 min and 1 h. Chlorine treatment was performed at 100, 250, 500, 1000, 3000 and 6000 ppm for 10 min. Samples exposed to 40 °C for 30 min or 1 h showed incidences of larvated eggs similar to controls. In contrast, no larvation occurred with eggs exposed to either 50 or 60 °C for 30 min or 1 h. Remarkably, in repeated assays, samples exposed to low doses of chlorine (100, 250, 500 and 1000 ppm for 10 min) showed significantly higher incidence of larvation than controls. Eggs treated with 3000 ppm for 10 min did not develop larvae, and no eggs were found after 6000 ppm treatment.     

Disinfection of Sparus aurata eggs with glutaraldehyde

This study was conducted to determine suitable conditions fordisinfecting eggs of gilthead sea bream (Sparus aurata)with glutaraldehyde. Effects of the developmental stage (4–8 cells,morula, blastopore closure or heart beating), of the concentration (C = 200,300or 400 ppm) and of the duration of the glutaraldehyde treatment (T= 2 to 10 min) were investigated. Before the blastopore closurestage, egg manipulation and treatment induced mortality. After this stage, thetoxicity of the glutaraldehyde treatment was negligible if the value of theproduct C × T was less than 1000. Above this value, the percentage ofhatching and of normal larvae decreased and the percentage of imprisoned larvaeincreased. Toxic effects of glutaraldehyde varied according to the egg qualityat the time of the treatment. It was concluded that 200 ppmglutaraldehyde for 4 min, at the blastopore closure stage or attheheart beating stage, were acceptable conditions for disinfecting gilthead seabream eggs at 18 °C.     

Disinfection of Sparus aurata eggs with glutaraldehyde

This study was conducted to determine suitable conditions fordisinfecting eggs of gilthead sea bream (Sparus aurata)with glutaraldehyde. Effects of the developmental stage (4–8 cells,morula, blastopore closure or heart beating), of the concentration (C = 200,300or 400 ppm) and of the duration of the glutaraldehyde treatment (T= 2 to 10 min) were investigated. Before the blastopore closurestage, egg manipulation and treatment induced mortality. After this stage, thetoxicity of the glutaraldehyde treatment was negligible if the value of theproduct C × T was less than 1000. Above this value, the percentage ofhatching and of normal larvae decreased and the percentage of imprisoned larvaeincreased. Toxic effects of glutaraldehyde varied according to the egg qualityat the time of the treatment. It was concluded that 200 ppmglutaraldehyde for 4 min, at the blastopore closure stage or attheheart beating stage, were acceptable conditions for disinfecting gilthead seabream eggs at 18 °C.     

Cryopreservation of sperm of common carp, Cyprinus carpio L.

Milt of common carp, Cyprinus carpio L. was cryopreserved in pellet form with the use of 12 extenders. Most efficient were: BE2 original extender (containing 85 mM NaCl, 50 mM KCl, 3 mm CaCl₂, 1 mm MgCl₂ with 10% dimethyl-acetamide (DMA) and 10% addition of hen's egg yolk) and Kurokura et al.'s extender with 15% DMA and 10% yolk (about 73% and 69% of eyed eggs, about 61% and 52% of swim-up larvae, respectively). Within the most effective treatments, survival from the eyed-egg stage to the swim-up stage was similar to that observed in the control group. Survival from the eyed-egg stage to the swim-up stage (percentage of eyed eggs was considered as 100%) was highly significantly and positively correlated with the actual rate of swim-up larvae.     

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.     

Prevention of Saprolegniasis in rainbow trout (Oncorhynchus mykiss) eggs using oregano (Origanum onites) and laurel (Laurus nobilis) essential oils

The present study investigated the antifungal effects of essential oils of oregano (Origanum onites) and laurel (Laurus nobilis) on Saprolegniasis, a disease that occurs in rainbow trout eggs during the incubation period. Oregano and laurel were ground after drying, and essential oils were obtained by water distillation method using a Clevenger apparatus. The essential oils were added to potato dextrose agar (PDA) at the rates of 1–1000 ppm, and the minimum inhibitory concentration (MIC) was determined as 250 ppm whereas the minimum lethal concentration (MLC) was determined to be 500 ppm for both plants. In the in vivo trials, fertilized eggs were treated with predetermined doses either by bathing during water hardening and incubation period or only during incubation period, and death rates were monitored during embryological development. The best larvae hatching rate was determined in 500 ppm oregano and 500 ppm laurel groups treated during water hardening plus daily as 82.11% and 79.87%, respectively. According to the results, it was determined that oregano and laurel essential oils exhibited better results in all doses compared with the negative control group, and 500 ppm dose had a better effect than the positive control group treated with formalin.     

三疣梭子蟹受精卵离体孵化技术

为了探究三疣梭子蟹受精卵的离体孵化技术及效果,本研究先后开展了受精卵块最适分离液种类及作用条件的筛选、分离液处理不同发育期受精卵离体孵化的差异、分离液处理对受精卵卵膜的结构影响,及分离液处理受精卵对孵化后幼体活力的影响实验。结果显示,木瓜蛋白酶是一种较为理想的分离液,在浓度为0.09 g/mL,分离时间30 min时,分离率可达到99%以上;经过分离液处理后的各期受精卵均能孵化出幼体,卵内溞状幼体期离体胚胎孵化率最高,为89.0%±3.3%,未经处理的对照组为70.0%±4.8%;卵裂期离体胚胎孵化率最低,为58.0%±3.9%,对照组为31.0%±2.3%,与对照组相比,处理组的孵化率明显提高。透射电镜结果显示,经过分离液处理的受精卵卵膜结构疏松,且厚度降低,符合处理组孵化率增加这一现象,干露、福尔马林溶液胁迫和行为学测试对不同处理组的幼体进行质量评价的结果显示,处理组和对照组幼体活力无显著差异。研究表明,实验所获得的分离液可以有效提高三疣梭子蟹受精卵的分离率和孵化率,且不影响幼体质量,可为三疣梭子蟹及其他甲壳动物受精卵的离体孵化提供参考。本研究可为三疣梭子蟹的苗种繁育提供新的技术手段,也将为基因编辑辅助育种等技术的实施奠定基础。     

The diet and growth of larval and juvenile pikeperch (Stizostedion lucioperca (L.)): A comparative study of fishponds and a reservoir

Diet of larval and juvenile pikeperch (Stizostedion lucioperca) reared in ponds wasinvestigated and compared with the diet ofpikeperch from a reservoir. The standard lengthof first feeding pikeperch larvae in ponds was6.1 mm, on average, and although rotifers werepresent in the diet, their numericalcontribution can be considered asinsubstantial. Rotifers were soon replaced bynauplii of cyclopoid copepods, which werehighly positively selected and contributedlargely to the diet up to a larval length of 10 mm.Daphnia spp. were consumed from the onsetof exogenous feeding, but were not positivelyselected until 15 mm. Anothersmaller cladoceran Bosmina longirostriswas highly negatively selected and did notcontribute significantly to the diet. A clearpositive selection for larger relatively tosmaller prey and a preference for Daphniafrom a body length of 15 mm onwards could beobserved. In the reservoir, rotifers were notfound in the diet of pikeperch larvae even inthe smallest individuals. Dominant food itemswere nauplii and 1st copepodite instar ofEudiaptomus gracilis and Cyclopsspp. Cladocerans – Daphnia galeata andto a lesser extent Diaphanosomabrachyurum appeared in the pikeperch diet at alength of about 10 mm. A shift from copepods toDaphnia spp. and especiallyLeptodora kindtii could be recognised inpikeperch at a length of 20 mm. When comparing ourdata from nursing ponds with the data fromímov reservoir, similar trends in dietcomposition were observed.Growth of pikeperch was found significantlyfaster in nursing ponds than in the reservoir.Slow growth of reservoir pikeperch was probablyan artefact due to the prolonged spawningperiod in the reservoir. Larvae and juvenilesfrom later spawnings decreased the average sizeof the population over the studied period. Innursing ponds lowest average standard length atharvest was found in the pond with the highestnumbers of fish, and vice versa in the pondwith the lowest numbers the largest standardlength was recorded. This result corresponds tothe increased intracohort food competitionamong juvenile pikeperch with increasingstocking density.     

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.     

The influence of triiodothyronine (T<Subscript>3</Subscript>) on the early development of piracanjuba (<Emphasis Type="Italic">Brycon orbignyanus</Emphasis>)

This paper reports the triiodothyronine’s (T₃) effects on the early growth and survival of piracanjuba (Brycon orbignyanus) produced from fertilized eggs hormone exposed. The study was carried out in two phases. In the first phase, eggs divided in 6 batches were immersed in T₃ solutions: 0.01; 0.05; 0.1; 0.5 ppm; 1 ppm and control (no T₃). After a 15-min immersion, eggs were transferred to incubators where larvae were kept up to 72 h after hatching. Larval weight, length and yolk sac volume were determined every 12 h. Sixty and 72 h after hatching, larvae exposed to 0.5 ppm T₃ were significantly heavier than the others, and those exposed to 1 ppm T₃ showed the lowest weight. The yolk sac absorption was not affected. In the second experimental phase, the resulting fry from the first phase were stocked into 3 boxes per treatment (5 larvae L⁻¹) and fed with plankton, fish larvae and feed prepared in the hatchery (48% CP) in the first 3 days, plankton and feed from the 4th to the 10th day and only feed in the next (last) 5 days. Fry weight, length and specific growth rate were determined at 1, 5, 10 and 15 days. Survival was calculated in the last day. In the 15th day, fry length did not differ among treatments but the weight of the control group was higher. Higher survival in the T₃-treated groups suggested lower predation among fry. The results allowed us to conclude that there was no expressive effect of T₃ on the growth, but it improved the survival of the piracanjuba progeny.     

Induced out-of-season spawning of the mud crab, Scylla paramamosain (Estampador) and effects of temperature on embryo development

Treated with combined bilateral eyestalk ablation and maintenance of water temperature at 22.5±1.5 °C, mud crab Scylla paramamosain females with mature ovaries were induced to produce eggs outside the natural spawning season in subtropical southern China. Newly extruded eggs from a crab were incubated in vitro at 10, 15, 20, 25, 27, 30, 35 °C, respectively, and the embryonic development was closely monitored. Abnormal cell division was observed at temperatures 10 and 35 °C. At 15 °C, development remained at the gastrula stage by day 32 post‐spawn, at which time the experiment was terminated. Hatching of in vitro incubated eggs occurred between 20 and 30 °C. An increase in incubation temperature from 20 to 25 °C reduced the incubation duration by 14 days, 2.6 times of that measured for a similar 5 °C increase from 25 to 30 °C. Embryonic development of S. paramamosain was divided into stage 0–10, and the duration of each stage was recorded for each incubation temperature. The information obtained allows accurate prediction of hatching time of female crabs incubated under variable temperatures. Larvae hatched from in vitro incubated eggs were reared to reach first juvenile crab stage and their dry weights were similar to those of larvae hatched naturally.