Involvement of Gonadotropin-Releasing Hormone in Thyroxine Release in Three different forms of Teleost Fish: Barfin Founder, Masu Salmon and Goldfish

Effects of gonadotropin-releasing hormone (GnRH) on thyroxine (T₄) release in vivo and in vitro were studied in barfin flounder Verasper moseri, masu salmon Oncorhynchus masou and goldfish Carassius auratus. Seabream GnRH (sbGnRH) at a dose of 200 ng/50 g body weight (BW) significantly increased plasma T₄ levels 1 h after the in vivo injection in the barfin flounder, but thereafter the levels normalized. Salmon GnRH (sGnRH) significantly increased plasma T₄ levels l h after the injection with a significant return to initial levels in male masu salmon and male goldfish. In contrast, sGnRH and cGnRH-II in barfin flounder, and cGnRH-II in male masu salmon and male goldfish were not effective in stimulating T₄ release. To clarify direct involvement of GnRH in T₄ release, dissected lower jaw including scattered thyroid follicles was incubated with sbGnRH (1 μg/well) in barfin flounder, and with two doses (0.1 and 1 μg/well) of sGnRH in masu salmon and goldfish in vitro. T₄ concentrations of control were stable during 24 h. Incubation of lower jaw with high dose (1 μg/well) of GnRH significantly (P<0.05) increased T₄ concentrations of incubation medium at 1 h in all experimental fishes. These results indicate that direct stimulation of T₄ secretion by GnRH occurs widely in teleost fish.     

Pharmacokinetics and tissue distribution of bronopol in grass carp,Ctenopharyngodon Idella,at 15 and 20°C

This study explored the pharmacokinetic characteristics of bronopol in grass carp (Ctenopharyngodon idella) under different temperatures. The concentrations of bronopol in the plasma and tissues (kidney, liver and muscle) of grass carp after soaking in bronopol (6.75 µg/ml) at 15 and 20°C were determined using liquid chromatography–mass spectrometry. The concentrations of bronopol in both plasma and tissue first increased and then decreased with soaking time at both tested temperatures. The peak bronopol concentrations at 15°C (plasma: 8.934 µg/ml; kidney: 9.23 µg/g; liver: 9.47 µg/g; and muscle: 7.98 µg/g) were slightly lower than those at 20°C (plasma: 9.654 µg/ml; kidney: 10.83 µg/g; liver: 13.40 µg/g; and muscle: 8.68 µg/g). In an analysis of pharmacokinetic parameters, the bronopol concentration–time profiles for plasma were best described by a two‐compartment, open pharmacokinetic model with first‐order absorption. The t_1/2β for plasma at 15 and 20°C was 82.933 and 92.041 hr, respectively, indicating that the elimination of bronopol was faster at 15°C than at 20°C. In addition, AUC_0‐t and AUC_0‐∞ values of bronopol were 477.892 and 495.809 µg/L hr at 15°C and 495.809 and 589.859 µg/L hr at 20°C, indicating that the content of bronopol in the organism was higher at higher temperature.     

Effects of Fish Size and Water Temperature on the Acute Toxicity of Copper for Japanese Flounder,Paralichthys olivaceus,and Red Sea Bream,Pagrus major

The acute toxicities of copper were examined for Japanese flounder, Paralichthys olivaceus, and red sea bream, Pagrus major, in terms of fish size and water temperature. Artificial seawater of low pH of 5.4–6.7 was used as testing water to keep dissolved copper concentration at 0.04–41 mg Cu/L. Japanese flounder of 0.3–17 g and red sea bream of 0.5–13 g were exposed to different concentrations of copper for 96 h at 20 C under semistatic condition. Median‐lethal concentration for 96 h of Japanese flounder and red sea bream were 8.7–12.2 and 2.0–5.2 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and fish size for Japanese flounder, while the value decreased significantly with increasing fish size for red sea bream. Effect of water temperature on the acute toxicity was examined for Japanese flounder of 0.3 and 0.4 g at 10, 15, 20, and 25 C and red sea bream of 0.5 and 1.0 g at 12, 15, 20, and 25 C. Ninety‐six‐hour median‐lethal concentrations for Japanese flounder and red sea bream were 5.1–11.2 and 1.0–5.3 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and water temperature for both fish species.     

Single Intravascular and Oral Dose Pharmacokinetics of Mebendazole in Blunt Snout Bream,Megalobrama amblycephala

Mebendazole (MBZ) is a broad‐spectrum benzimidazole methylcarbamate anthelmintic used widely in animal husbandry and aquaculture. However, there is no information available on the pharmacokinetic behavior of MBZ in blunt snout bream, Megalobrama amblycephala. In this study, pharmacokinetic parameters of MBZ were estimated in blunt snout bream after intravascular (3 mg/kg body weight [BW]) and oral (20 mg/kg BW) administration. The analyses of plasma samples were performed using ultra performance liquid chromatography with ultraviolet detector. After intravascular administration, plasma concentration–time curves were best described by a two‐compartment open model. The distribution half‐life (t_1/2α), elimination half‐life (t_1/2β), and area under the concentration–time curve (AUC) of blunt snout bream were 0.1 h, 27.9 h, and 56666.0 h.µg/L, respectively. After oral administration, a one‐compartment open model with first‐order absorption best fit the plasma data. The absorption half‐life (t_1/2Ka), elimination half‐life (t_1/2Ke), peak concentration (C_max), time‐to‐peak concentration (T_max), and AUC of blunt snout bream were estimated to be 1.9 h, 34.6 h, 918.1 µg/L, 8.4 h, and 54201.4 h.µg/L, respectively. The oral bioavailability (F) was 14.3 %. The pharmacokinetics of MBZ in blunt snout bream displayed low bioavailability, relatively rapid absorption, and relatively rapid elimination.     

Toxicity of Hydrogen Peroxide to Fingerling Walleyes

The toxicity of hydrogen peroxide (H₂O₂) to fingerling (80-160 mm total length) walleye, Stizostedion vitreum, was evaluated. Walleye were exposed in a 1 hour static bath to 50, 75, 100, 200, 225, 250, 400, and 600 ppm (µL/L) active ingredient (H₂O₂) in five experiments. Three to six replicates of each concentration were used in each experiment, with 25 fish per replicate. After resuming flow, concentrations of H₂O₂ in the tanks declined by 23.8% every hour, reaching 0 ppm (µL/L) about 3.5 hours after treatment. Mortalities, as well as dissolved oxygen, temperature, and H₂O₂ concentrations were monitored at 1, 12, 24, 48, 72, and 96 hours. Dissolved oxygen concentrations increased 0.25-1.16 ppm (µL/L) 1 hour after the addition of H₂O₂, because of the dissociation of H₂O₂ (2H₂O₂→2H₂O + O₂). The LC₅₀ following a single 1-hour exposure ranged from 145.1 ppm (µL/L) at 12 hours to 142.8 ppm (µL/L) at 96 hours, which indicates that the toxic effect is mainly from initial contact. The toxicity threshold for walleye is about 50 ppm (µL/L) (1.3% mortality at 96 hours following 1 hour exposure). Other studies indicate that rainbow trout, Oncorhynchus mykiss, tolerate up to 250 ppm (µL/L) for 1 hour, but fingerling walleye should not be exposed to more than 50 ppm (µL/L) for the same exposure interval.     

Effects of fish size and water temperature on the acute toxicity of boron to Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> and red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:     The acute toxicities of boron were examined for Japanese flounder Paralichthys olivaceus and red sea bream Pagrus major in terms of fish size and water temperature. Japanese flounder of 0.1–70.0 g and red sea bream of 0.6–20.3 g were exposed to different concentrations of boron for 96 h at 20°C under semi-static conditions. In both fish species, the median lethal concentration (LC₅₀) for 96 h of boron increased linearly with increasing fish size, ranging from 108 to 252 mg B/L for the flounder, and from 97 to 172 mg B/L for the sea bream. The effect of water temperature on acute toxicity was examined for Japanese flounder of 0.6 and 1.5 g at 10, 15, 20 and 25°C, and for red sea bream of 0.6 and 2.4 g at 12, 15, 20 and 25°C. The toxicity of boron for the flounder increased linearly with increasing water temperature. The 96 h LC₅₀ values ranged from 299 to 108 mg B/L for the 0.6 g flounder and from 350 to 113 mg B/L for the 1.5 g flounder. A similar trend was shown for the 2.4 g red sea bream; however, the relationship for the 0.6 g red sea bream was not significant.     

Nitrite Toxicity and Methemoglobin Changes in Southern Flounder,Paralichthys lethostigma,in Brackish Water

This study was performed to estimate the nitrite toxicity to southern flounder, Paralichthys lethostigma, in brackish water (7.5 ppt of salinity). For a LC₅₀ test, 20 fingerlings (5.7 ± 0.4 cm) in each aquarium (15 L) were exposed to the concentrations of 0, 1, 5, 10, 15, 30, 60, 120, and 240 mg NO₂^?‐N/L in duplication for 10 d. Median lethal concentration at 96 h (96‐h LC₅₀) was calculated as 81.6 mg NO₂^?‐N/L. For a verification test, young flounder (164.2 ± 9.1 g) were exposed to a simulated culture condition in recirculating systems (1000 L). Sodium nitrite was not added to control system, whereas it was added to Treatment system 1 (TS 1) and Treatment system 2 (TS 2) to maintain nitrite concentrations of 20 and 30 mg NO₂^?‐N/L, respectively. The plasma nitrite concentrations of the young flounder in TS 1 and TS 2 were 4.5 and 6.6 mg NO₂^?‐N/L, respectively, after 2 wk. At this time, the methemoglobin percentages in TS 1 and TS 2 reached 85.8 and 89.7%, and survival rates were 37.5 and 25.0%, respectively. The results of these tests indicate that southern flounder do not concentrate nitrite in blood from the environment, but they seem to be more sensitive to nitrite compared with other species that do not concentrate nitrite.     

Effect of Rosmarinus officinalis Essential Oil and Nisin on Streptococcus iniae and Lactococcus garvieae in a Food Model System

The effects of concentrations of Rosmarinus officinalis essential oil (EO) and nisin (N) as well as temperature and storage time on growth of Streptococcus iniae and Lactococcus garvieae in Oncorhynchus mykiss were evaluated. According to analysis of the rosemary EO, the 1, 8-cineol and α-pinene were the predominant components. The growth of S. iniae was significantly decrease by EO concentrations at 4 ºC. For L. garvieae, the viable count was significantly inhibited by EO and N singly and in combinations, incubated at bath storage of 0.25 and 0.5 µg/mL proved insufficient to act against S. iniae and L. garvieae. The combinations of the rosemary EO at 0.0015% with N at 0.5 µg/mL showed stronger antimicrobial effect against two bacterial than the rosemary EO at 0% but lower than the combination with N at 0.5 µg/mL and EO at 0.045% which in turn was lower than of the rosemary EO at 0.135%. In its turn, rosemary EO showed lower antimicrobial activity than its combinations with N, which showed a bactericidal effect against the pathogens. The best inhibitory effects of EO in combinations with N for two bacterial were obtained at combinations of EO=0.135% and N=0.5 µg/mL.     

The effects of different proportions of the 17β‐estradiol and 17α‐methyltestosterone on growth,sex reversal and skin colouration of the electric blue hap (Sciaenochromis ahli Trewavas, 1935)

This study was performed to investigate the effects of 17β‐estradiol (ES) and 17α‐methyltestosterone (MT) on growth, development, survival, sex ratio and colour change in the electric blue hap (Sciaenochromis ahli Trewavas, 1935). The hormones were not supplemented to the control feed, while six other feeds were prepared by adding 20, 40 and 60 mg kg^?1 17β‐ES or 20, 40 and 60 mg kg^?1 17α‐MT to each, resulting in seven different feed treatments. Average live weight of the fish supplemented with these diets was 0.42 ± 0.04 g. At the end of the study, the highest weight gain was observed in fish fed 60 mg kg^?1 17α‐MT group (2.62 ± 0.11 g) and the difference with the groups fed with 17β‐ES was found to be significant. All fish fed 17α‐MT were male, while the rates of feminization in fish fed 17β‐ES at 20, 40, 60 mg kg^?1 were 91.11%, 88.88% and 93.33% respectively. Survival rates were respectively determined as 80%, 95.56%, 84.44%, 93.33%, 77.78%, 84.44% and 84.44% for the control, 20, 40, 60 mg kg^?1 17β‐ES and 20, 40, 60 mg kg^?1 17α‐MT treatments. The best colouration was achieved in the 17α‐MT groups (P < 0.05). The L* values varied between 32.98 ± 4.44 and 61.35 ± 2.19, a* values between ?7.06 ± 0.22 and ?3.42 ± 0.11, and b* values between ?7.74 ± 0.10 and 11.65 ± 0.03, while Chroma (C*) and Hue (H°_ab) angle values varied between 7.54 ± 0.22 and 13.60 ± 0.01 and between 119.76 ± 0.05 and 239.73 ± 4.86. In conclusion, the 17α‐MT feeding was found to have a greater effect on the growth, feed conversion ratio, masculunization and pigmentation of the electric blue haps than the 17β‐ES treatment.     

Hg2+对克隆牙鲆和普通牙鲆的急性毒性

以相同规格的牙鲆Paralichthys olivaceus普通群体为对照,采用静水生物测试法开展了Hg2+对牙鲆克隆群体的急性毒性实验,实验所用幼鱼为130日龄,体长为(12.38±1.13)cm,体质量为(17.97±5.79)g。实验期间,水温为(21±1.2)℃,pH为8.1-8.3,盐度为31.0,硬度6200 mg/L,连续充气保证溶氧充足。研究表明,实验初始阶段,高浓度组克隆牙鲆在容器内躁动不安,上升到水面沿池壁缓慢转圈游动,一段时间后安静伏在池底;而普通牙鲆则仅个别鱼在水面游动,多数静止在池底不动。随中毒程度加深,用玻璃棒触碰普通牙鲆时,其反应剧烈,在水中侧翻、打转、急速游动或上下窜动,而克隆牙鲆则仅沿池底缓慢游动。最终克隆牙鲆和普通牙鲆均身体僵直,失去呼吸能力死亡。Hg2+对克隆牙鲆的24、48、96 h的半致死质量浓度分别为1.50、0.86、0.84 mg/L,而对普通牙鲆的半致死质量浓度则分别为1.93、1.04、0.90 mg/L。结果表明,克隆牙鲆对Hg2+的耐受性弱于普通牙鲆,对Hg2+更加敏感。从死亡过程看,各处理组的克隆牙鲆从出现死亡个体开始,死亡时间集中在8 h内,具有同步性;普通牙鲆则在实验期间陆续出现死亡,持续时间较长,同步性较差。     

Short‐term Effects of Genistein on the Reproductive Characteristics of Male Gibel Carp,Carassius auratus gibelio

The objective of this study was to determine the effects of genistein or 17β‐estradiol (E2) on the reproductive physiology in male gibel carp, Carassius auratus gibelio. Maturing male gibel carp received intraperitoneal injections of E2 (10 µg/g body weight), one of two genistein doses (5 µg/g body weight, G5, or 50 µg/g body weight, G50), or the injection vehicle every other day for 10 d. Disruptions in reproductive capacity were determined by measuring indices of sperm quality, plasma metabolites and sex steroids, histological analyses of testes, fertilization rate, and offspring viability. E2 and genistein treatment reduced gonadosomatic index and milt volume, while reduction in spermatozoa concentration and spermatocrit occurred only in E2 and G50‐treated males. Histological examination of the testes indicates that E2 and genistein inhibited reproductive capacity through disruption of the spermatogenesis in males. Genistein reduced fertilization rate and offspring viability at 6 d after hatch (d.a.h.). Plasma testosterone and E2 decreased and increased, respectively, with E2 and G50 treatment. E2 and G50 treatment altered plasma metabolite phosphorus, calcium, cholesterol, and triglyceride. These findings indicate that genistein can negatively affect reproductive capacity in male gibel carp, suggesting that high dietary genistein may impair gonad development.     

Pharmacokinetics of Dietary 13C-labeled Icosapentaenoic Acid in Japanese Flounder Paralichthys olivaceus

The objectives of this study were to investigate: 1) the pharmacokinetics of dietary icosapentaenoic acid (EPA) in the plasma using ¹³C‐labeled EPA ([¹³C]EPA); 2) the bioavailability of [¹³C]EPA with different chemical forms; and 3) an effective plasma EPA level for normal growth of the Japanese flounder Parachthys olivaceus. [¹³C]EPA was biosynthesized using the Chlorella Nannochloropsis oculate, and 1‐myristoyl 2‐[¹³C]icosapentaenoyl phasphatidyl‐choline ([¹³C]EPA‐PC) and [¹³C]EPA ethyl ester ([¹³C]EPA‐EE) were chemically synthesized from [¹³C]EPA. Free [¹³C]EPA ([¹³C]EPA‐FREE) dissolved in 0.5% tragacanth gum‐polye‐thyleneglycol (3:1, V/V) was administered to the Japanese flounder at dosages of 2.8, 5.6, and 16.8 mg/kg by a single oral administration. [¹³C]EPA‐PC (44.8 mg/kg) and [¹³C]EPA‐EE (18.3 mg/kg), equimolar to 3.0 mg [¹³C]EPA‐FREE (16.8 mg/kg), were administered to the fish in a similar manner. In [¹³C]EPA‐FREE dosed fish, the kinetics of the mean plasma [¹³C]EPA were linear with respect to dose. [¹³C]EPA‐FREE and [¹³C]EPA‐PC were more efecient in maintaining high plasma EPA levels than [¹³C]EPA‐EE in the Japanese flounder. [¹³C]EPA was distributed in plasma, blood cell, eyeball containing the orbital fat, liver, stomach, intestine, skin, brain, heart and muscle in three [¹³C]EPA derivatives studies. The effective plasma EPA level for normal growth of the Japanese flounder is estimated to be 307 to 937 μg/mL for EPA‐FREE administration, and from 286 to 815 μg/mL 6.3 to 38 mg/mL for the EPA‐PC administration.     

Pharmacokinetic/pharmacodynamic properties and P‐glycoprotein expression in pacific white shrimp,Litopenaeus vannamei after oral administration at three dosages of oxolinic acid

The experiments explored the pharmacokinetics (PK) properties of oxolinic acid (OXA) after oral administration at three dosages (10, 30 and 80 mg/kg) via medicated feed in the shrimp. The results showed that the C_max values of 4.31, 14.93 and 16.62 mg/L and AUC_0–∞ values of 92.61, 252.30 and 364.27 mg hr^?1 L^?1 were observed at three OXA dosage groups in the haemolymph respectively. In the hepatopancreas, C_max values of 7.90, 27.23 and 60.51 mg/kg and AUC_0–∞ values of 42.01, 133.06 and 219.06 mg hr^?1 L^?1 were observed at 0.5 hr post administration respectively. In the muscle, C_max values of 1.62, 5.80 and 7.36 mg/kg and the AUC_0–∞ values of 25.64, 98.10 and 134.24 mg hr^?1 L^?1 were observed at 2 hr post administration respectively. In the gills, C_max values of 2.87, 8.08 and 12.12 mg/kg and the AUC_0–∞ values of 51.38, 118.65 and 206.48 mg hr^?1 L^?1 were observed at 4 hr post administration respectively. In addition, the in vitro MIC values of OXA at three dosages against 132 strains of Vibrio were examined and showed that the minimum inhibitory concentration (MIC) values for OXA primarily ranged from 0.15–1.25 µg/ml, including eight strains of Vibrio showing MIC values ≥5 µg/ml. The MIC₅₀ and MIC₉₀ values of 132 strains were 0.62 and 1.25 μg/ml respectively. The AUC_0–24/MIC₉₀ ratios of Vibrio were 140.4 in 30 mg/kg group. Furthermore, the P‐glycoprotein (P‐gp) expression was determined in shrimp tissues after administration to three dosage groups (10, 30 and 80 mg/kg). The results showed that P‐gp expression was up‐regulated in the hepatopancreas (5.36‐, 13.68‐ and 31.06‐fold respectively) compared with the control group.     

Effects of photoperiod on gonadotropin-releasing hormone levels in the brain and pituitary of underyearling male barfin flounder

ABSTRACT:   A pleuronectiform fish, the barfin flounder Verasper moseri , expresses three gonadotropin-releasing hormone (GnRH) forms in the brain: salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II) and seabream GnRH (sbGnRH). To clarify the effects of photoperiod on GnRH systems, changes in brain and pituitary GnRH peptide levels were examined using time-resolved fluoroimmunoassays. In experiment 1, 5-month-old male barfin flounder (mean total length 9.0 cm, body weight 11.0 g) were divided into short (8:16 h light : dark [L:D] cycle; lights on 08.00–16.00 hours) and long photoperiod (16:8 h L:D cycle; lights on 04.00–20.00 hours) groups in mid September and maintained until November under natural water temperature (19.3–15.2°C). Brain sGnRH concentrations were significantly higher in the 16:8 h L:D group than in the 8:16 h L:D group, whereas no significant differences were observed in total length, body weight, plasma testosterone concentration, brain cGnRH-II concentration and pituitary sbGnRH content. In experiment 2, 7-month-old male barfin flounder (mean total length 16.5 cm, body weight 76.8 g) were divided into short and long photoperiod groups in mid December and maintained until February under natural water temperature (12.5–6.6°C). Total length, body weight and condition factor were significantly greater in the 16:8 h L:D group than in the 8:16 h L:D group, whereas no significant differences were observed in plasma testosterone concentration and GnRH levels in the brain and pituitary. These results indicate that levels of sGnRH in barfin flounder are influenced by photoperiodic treatment dependent on water temperature and/or body size.     

Anaesthetic effect of eugenol at different concentrations and temperatures on black tiger shrimp (Penaeus monodon)

The anaesthetic effects of eugenol on Penaeus monodon were investigated at the different eugenol concentrations (60, 110, 160 and 210 mg/L), water temperature (21, 26 and 31°C), air exposure time (3, 6, 9 and 12 min) and body weight (2.62 ± 0.27, 6.34 ± 0.36 and 11.43 ± 0.33 g). The anaesthesia and recovery time were recorded. The results showed that the anaesthesia time of the shrimp decreased with the increase in the eugenol concentration and water temperature, and the recovery time increased with the increase of the eugenol concentration and the decrease of water temperature. Under the same eugenol concentrations, the recovery time increased with the increase of air exposure time and body weight. Under the eugenol concentration range of 60–210 mg/L, the recovered rate was 100%. The results indicated that eugenol is a safe and efficient anaesthetic for P. monodon.     

Pharmacokinetics and physio‐metabolic response of single and multiple dose of fenbendazole in Labeo rohita (Hamilton, 1822) fingerlings

An experiment was carried to determine the plasma fenbendazole (FBZ) concentration and physio‐metabolic responses in juveniles of Labeo rohita (90 ± 4 g) after oral administration of single doses at 10, 20 and 50 mg, 20 mg FBZ/kg b.wt. in multiple times on 1st, 3rd and 7th day. The blood samples were collected at 0.5, 1, 2, 4, 8, 12, 24, 30, 48, 72, 96 and 120 hr, after single‐dose administration, and regularly (upto 15 day) in multiple dose. Plasma FBZ concentration was determined up to the limit of detection (LoD) of 0.09 µg/ml by HPLC. There was no parent drug detected in plasma for administration of 10 mg FBZ/kg b.wt. The drug attained the peak concentration (C_max) 1.85 and 3.09 µg/ml in plasma at 4 hr (T_max) after administration of 20 and 50mg FBZ/kg b.wt. respectively. Plasma FBZ was detectable up to 96 and 120 hr with concentration 0.09 ± 0.007 and 0.098 ± 0.006 µg/ml, respectively, after single‐dose administration of 20 and 50mg/kg b.wt. In case of multiple‐dose administration, the maximum concentration of FBZ was 1.01 ± 0.03 µg/ml on 7th day that was less than to the single dose at 50 mg/kg b.wt. However, FBZ was detected up to 11 day after multiple doses. The study revealed that the hepatic antioxidant enzymes activities like superoxide dismutase, catalase and glutathione‐S‐transferase were significantly affected by increasing FBZ in single and multiple doses. The results of the present study could reveal that single‐ or multiple‐oral administration of FBZ at 20 mg/kg b.wt. in feed as antihelminthic drug in L. rohita could be considered as the safe dose.     

Anaesthetic efficacy of eugenol on various size classes of angelfish (Pterophyllum scalare Schultze, 1823)

Anaesthetic efficacy of eugenol was investigated on Pterophyllum scalare. A total of 130 fish with average weights of 1.0 ± 0.5, 5.0 ± 1.0 and 10.0 ± 1.0 g were subjected to 1.25, 2.5, 4.0, 5.5 and 7.0 mg/L eugenol, and behavioural responses were observed. Induction and recovery times were significantly affected by the interactive effect of eugenol concentration and fish weight (p < .05). Generally, 49.9–128 s after exposure to 1.25–7 mg/L eugenol, fish reached stage 3. Fish entered stage 4 over 55–135 s post exposure to such concentrations. Recovery time was 393.5–597.7 s in all sizes. Any increase in eugenol concentration led to a significant decrease in the induction time with a subsequent increment of the recovery time. Concentrations of eugenol and fish size along with their interactive effects have significantly contributed to the regression models, with concentration recording the highest beta values for stages 1, 2, 3 and 4 (?0.903, ?0.898, ?0.976 and ?0.864 respectively) and the product of size and anaesthetic concentration for full recovery in smaller fish (0.647) and eugenol concentration in larger ones (0.967). Recovery time was fitted to induction time to stage 4 via quadratic and linear regression models in smaller and larger fish respectively. Results revealed the minimal eugenol concentration to induce anaesthesia in various size classes of angelfish in less than 3 min was 1.25 mg/L. Our results showed eugenol as an effective and safe anaesthetic; however, it is not advisable for live fish transportation.     

Protective Effect of Emodin against Hyperthermia‐induced Stress in Hepatic Cells of Grass Carp,Ctenopharyngodon idellus

Emodin is an anthraquinone exhibiting several positive benefits of anti‐inflammatory, free radical scavenging, and resistance to stress. The goal of this study is to investigate the effects of emodin on the hepatic cell line of grass carp exposed to hyperthermic stress. Cultured cells were treated with various emodin concentrations (0, 0.04, 0.2, and 1.0 µg/mL) at 27 C for 24 h. Then all cultured cells were exposed to thermal stress by increasing the culture temperature (32 ± 0.5 C) for 0.5 h. Increased temperatures significantly reduced cell viability and increased lactate dehydrogenase (LDH) release in three of the four experimental groups (0, 0.2, and 1 µg/mL emodin) compared with the control. Additionally, reactive oxygen species (ROS) were significantly higher in cells exposed to elevated temperatures and treated with 0.2 or 1 µg/mL emodin and mitochondrial membrane potential was significantly lower in cells treated with 0, 0.2, or 1 µg/mL emodin. Expressions of heat shock proteins (HSP60, HSP70, and HSP90) were significantly higher in all but one of the experimental groups. Our results suggest that 0.04 µg/mL emodin can increase cell viability and HSP90 mRNA level, reduce LDH release and concentration of ROS, and contribute to enhance the resistance to high temperature stress in the hepatic cells of grass carp.     

Effect of Hybrid Abalone,Haliotis discus hannai × Haliotis discus discus,Cultivation on the Carbon Cycle: Carbon Source/Sink

Respiration, calcification, and bio‐deposition of hybrid abalone, Haliotis discus hannai × Haliotis discus discus, fed on different foodstuffs have been measured to evaluate the effect of hybrid abalone culture on carbon source/sink in coastal areas. Fed with Laminaria japonica, Undaria pinnatifida, Gracilaria lemaneiformis, U. pinnatifida, and Ulva pertusa, alternated mutually, the carbon bio‐deposition rate of hybrid abalone was 24.29 ± 6.39, 65.40 ± 10.55, 21.48 ± 5.99, and 29.28 ± 6.47 µg/g/h, respectively. Hybrid abalone fed on U. pinnatifida had a higher carbon bio‐deposition rate compared to that fed on other foodstuff (P < 5%). Rate of CO₂ released by respiration of hybrid abalone fed on the experimental foodstuff was 24.53 ± 8.57, 32.73 ± 7.99, 29.31 ± 6.39, and 33.67 ± 12.37 µg/g/h, respectively. Results indicated that calcification presented less relationship with body weight type of the foodstuff. The rate of CO₂ released by calcification into seawater and atmosphere was 2.77 ± 1.89 and 6.53 ± 3.36 µg/g/h, respectively. The total rate of CO₂ released because of bio‐deposition, respiration, and calcification processes was 16.19 ± 4.67 µg/g/h, while the total rate of carbon sequestered in shells and tissues was 8.94 ± 2.07 µg/g/h. The study revealed that hybrid abalone culture is a source of CO₂.     

Diel Variation in RNA/DNA Ratios of Japanese Flounder Paralichthys olivaceus

Laboratory-reared Japanese flounder Paralichthys olivaceus larvae (11.2 ± 0.2 mm total length; age 26 d) were sampled over a 48-h period to determine any diel patterns in RNA/DNA ratios. RNA/DNA ratios were highest during daytime periods (0800, 1100, 1400, 1700, 2000 h) and significantly reduced at night (2300, 0200, 0500 h). Findings from this study indicate a diel variation in biochemical condition and suggest that special care should be taken in sampling design and data analysis to account for this source of variability.