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.     

Change in blood stress and antioxidant markers and hydromineral balance of common carp (Cyprinus carpio) anaesthetized with citronellal and linalool: Comparison with eugenol

The present study aimed to investigate the effects of anaesthesia with citronellal and linalool on stress, antioxidant and hydromineral responses of common carp. Eugenol was used as control anaesthetic, as it is a common anaesthetic in aquaculture. The fish (110 ± 5.65 g) were exposed to 406 mg/L citronellal, 982 mg/L linalool or 43 mg/L eugenol within 5 min (short‐term anaesthesia) or 10 and 20 mg/L of each anaesthetic for 3 hr (long‐term exposure), before blood sampling. The results showed that the short‐term anaesthesia had no significant effects on plasma lactate, sodium, chloride and phosphorus levels and blood catalase (CAT) and superoxide dismutase activities. The short‐term anaesthesia with linalool led to higher stress responses (increased cortisol and glucose levels) and oxidative stress (increased malondialdehyde [MDA] level and decreased total antioxidant capacity [TAC] and glutathione peroxidase [GPx] activity) compared to the eugenol and citronellal. Citronellal group had significantly higher cortisol and lower TAC level and GPx activity compared to eugenol group. Under the long‐term exposure, increase in anaesthetic concentration led to significant increase in plasma cortisol, glucose and phosphorus levels. The linalool‐treated fish had significantly higher cortisol and MDA levels compared to the other groups. The eugenol group had significantly higher lactate and phosphorus and lower TAC levels compared to the other groups. Although citronellal causes slight increase in plasma cortisol level, it induces lower tissue damages compared to eugenol. Thus, citronellal might be an alternative anaesthetic for carp anaesthesia based on the present tested parameters.     

丁香酚对花鲈幼鱼的麻醉效果

为研究丁香酚对花鲈(Lateolabrax maculatus)幼鱼的麻醉效果。采用静水方法在水温(27±1)℃下研究不同质量浓度丁香酚(20、25、30、40、50、60、80 mg/L和100 mg/L)对规格为(21.6±2.75)cm长和(110.3±30.67)g重的花鲈麻醉效果和呼吸频率的影响。25~100 mg/L质量浓度的丁香酚均能使花鲈进入深度麻醉期,且麻醉浓度与平均入麻时间呈负相关,与平均复苏时间成正相关;丁香酚质量浓度为50 mg/L时,麻醉效果较好。低浓度丁香酚20 mg/L对花鲈呼吸频率影响不明显。在质量浓度达到40~100 mg/L时,鱼体由麻醉期(A3)进入深度麻醉期(A4),呼吸频率迅速降低。以50 mg/L丁香酚将花鲈麻醉后再在空气中进行暴露,暴露时间与复苏时间呈正相关。暴露时间在2~30 min范围内的复苏率为100%。当时间增加至35、40和45 min时,复苏率降低至66%,33%和0。且暴露时间大于20 min花鲈只能进入恢复期3期(R3)。因此,50 mg/L的丁香酚麻醉液是花鲈幼鱼的理想麻醉浓度。     

Anaesthetic efficacy of eugenol on Flowerhorn (Amphilophus labiatus × Amphilophus trimaculatus)

Anaesthetic efficacy of eugenol was investigated on Flowerhorn (Amphilophus labiatus × Amphilophus trimaculatus). A total of 104 fish with average weights of 12 ± 2.5, 28 ± 5 and 53 ±5.1 g were subjected to 25–200 mg L^?1 eugenol and behavioural responses as well as induction and recovery times were recorded. Induction and recovery times were significantly affected by eugenol concentration as well as fish weight (P < 0.05). Generally, 49.9–127.3 s after exposure to 50–200 mg L^?1 eugenol, fish reached stage 3 anaesthesia (suitable for general handling). Fish entered stage 4 anaesthesia (suitable for surgery and blood sampling) over 57.3–140.4 s post exposure to such concentrations. Recovery time was 91.7–312 s in all weight classes for all eugenol concentrations. Mortality (23%) was only observed in 12‐g fish when were subjected to 200 mg L^?1 eugenol. This study showed the behavioural response of Flowerhorn to anaesthesia and eugenol efficacy as an anaesthetic in this important ornamental species. The general quadratic equation revealed that concentrations of eugenol and fish size along with their interactive effects have significantly contributed to the model, with concentration recording the highest beta value in all models (β = ?0.809, ?0.818 and ?0.909, P = 0.000). According to the results, minimum eugenol concentration to induce anaesthesia in less than 3 min was 50 mg L^?1.     

Alpinia galanga oil—A new natural source of fish anaesthetic

This study explores the anaesthetic activity of Alpinia galanga oil (AGO) in fish. Cyprinus carpio (koi carp) was used as a fish model. It was found that the induction time to stage 3 anaesthesia and the recovery time of the fish after exposure to AGO were exponentially and polynomially correlated to AGO concentrations. The viability of normal blood cells of koi carp anaesthetized with 500 mg/L AGO was found to be higher than 90% for normal red blood cells and white blood cells and 89% for peripheral blood nuclear cells indicating nontoxicity of AGO to the fish. A concentration of 300 mg/L of AGO was the most suitable for anaesthetizing koi carp due to the safety and effectiveness aspects as being ideally fitted to anaesthetic criteria. This concentration gave the induction time of 205.55 ± 5.07 s and the recovery time of 202.50 ± 9.30 s. Determination of stress biomarker such as blood cortisol and glucose as well as gene expression showed that the blood cortisol level of the fish anaesthetized with AGO was similar to normal levels. Moreover, blood glucose level was significantly less increased than those anaesthetized with tricaine methanesulfonate. Gene expressions of the fish cortisol receptor, cytochrome oxidase subunit1, heat shock protein 70 and Na⁺/K⁺‐ATPaseα₃ were significantly reduced after exposure to AGO indicating the advantages of AGO on fish stress reduction. Thus, AGO is a promising natural source for an alternative fish anaesthetics.     

Anaesthetic efficacy of eugenol on iridescent shark,Pangasius hypophthalmus (Sauvage, 1878) in different size classes

Anaesthetic efficacy of eugenol was investigated on iridescent shark, Pangasius hypophthalmus. Fish (2, 5, 10 and 20 g) subjected to 20–200 mg L^?1 eugenol and behavioural response as well as induction and recovery times were recorded. Induction and recovery times were significantly affected by eugenol concentration as well as fish weight (P < 0.05). Generally, 27–300 s after exposure to 20–200 mg L^?1 eugenol, iridescent sharks reached stage 3 anaesthesia (suitable for general handling). Fish entered stage 4 anaesthesia (suitable for surgery and blood sampling) over 54–710 s exposure to such concentrations. Recovery time was 109–600 s in all weight classes as well as eugenol concentrations. Mortality (44–100%) was only observed in 2 g fish when subjected to 110–170 mg L^?1 eugenol. This study, for the first time, showed behavioural response of iridescent shark to anaesthesia as well as effectiveness of eugenol as anaesthetic in this important aquaculture‐ornamental species. According to the models obtained in this study, minimum eugenol concentrations to induce anaesthesia over less than 3 min were 53.8–81.5 mg L^?1 in 2–20 g fish. Likewise, maximum eugenol concentrations in which fish recovered over less than 5 min were 65.9–105.8 mg L^?1 in 2–20 g fish.     

Menthol as anaesthetic for lambari Astyanax altiparanae (Garutti & Britski 2000): attenuation of stress responses

The anaesthetic potential of menthol was evaluated in lambari Astyanax altiparanae by exposing fingerlings to concentrations 50, 100, 150, 200, 250 and 300 mg L^?1 and measuring the induction and recovery times to deep anaesthesia, the mortality rates during and 96 h after procedure and after 6 min of continuous exposure. The effect of menthol on stress responses were evaluated by comparing glucose and cortisol levels of juveniles subjected to anaesthesia (50 mg L^?1), stress (air exposure) or pre‐anaesthesia associated to stress. All concentrations induced deep anaesthesia within 0.5 to 1 min, with recovery between 1.83 and 4.16 min, without mortality during the induction or up to 96 h after exposure. Induction time decreased and recovery time increased linearly as the menthol concentration increased. Continuous exposure to 50, 100 and 150 mg L^?1 concentrations resulted in mortality rates of 0%, 20% and 80% respectively. Anaesthesia or air exposure increase blood glucose but prior anaesthesia with menthol suppressed the elevation of cortisol caused by stress. Menthol has an anaesthetic effect and attenuates the stress response in lambari and 50 mg L^?1 is the most effective concentration for inducing deep anaesthesia in 1.0 min, safe for up to 6 min exposure.     

Minimally invasive evaluation of the anaesthetic efficacy of MS‐222 for ornamental discus fish using skin mucus biomarkers

Skin mucus has been demonstrated to provide stress biomarkers for evaluating the physiological status, providing new convenient and non‐invasive methods to detect stress response in fish. Here, we investigated the anaesthetic efficacy of tricaine methanesulphonate (MS‐222; 75–115 mg/L) for discus Symphysodon aequifasciata (34.27 ± 4.46 g; 8.10 ± 0.59 cm) using skin mucus stress biomarkers. The induction time, recovery time and respiratory frequency were also determined. According to the criteria for anaesthesia and recovery, discus fish to reach stage A3 (deep anaesthesia) within 3 min and to reach stage R4 (full recovery of normal behaviour) within 5 min were observed at 95–105 mg/L MS‐222. Respiratory frequency increased first and then decreased during MS‐222 exposure and increased after recovery. At 10 min after deep anaesthesia, a lower mucus glucose was only observed at 115 mg/L MS‐222. No change in mucus cortisol and increased lactate were observed in all treatments. Increased mucus protein was observed at 75, 85 and 95 mg/L MS‐222. At 10 min after recovery, increased mucus glucose and decreased mucus protein were observed at 85, 95 and 115 mg/L MS‐222, but increased mucus cortisol only at 115 mg/L and lactate only at 75 and 105 mg/L MS‐222. At 24 hr after recovery, mucus glucose returned to the initial level only at 75, 95 and 105 mg/L MS‐222, while cortisol at 75 and 85 mg/L and protein and lactate at 75 mg/L respectively. Overall, the effective dose of MS‐222 for discus fish has been suggested to be 95–105 mg/L.     

The efficacy of MS‐222 as anaesthetic agent in four freshwater aquarium fish species

The efficacy of anaesthetic tricaine methanesulfonate (MS‐222) was evaluated in four freshwater aquarium fish species, Zebrafish (Danio rerio), Guppy (Poecilia reticulata), Discu (Symphysodon discus) and Green swordtail (Xiphophorus helleri). The correct dose of anaesthetic should induce the plane 4 of anaesthesia in less than 180 s, recovery in less than 300 s and must survive when exposed during 30 min to anaesthetic. Fishes were exposed to six concentrations of anaesthetic (75, 100, 125, 150, 200 and 250 mg L^?1) and the time of fish reaching plane 4 of anaesthesia, post exposure recovery, and the percentage of survival when fish were subject to 30 min in the anaesthetic were recorded. The optimal doses varied according to the species: D. rerio – 75, 100 and 125 mg L^?1, P. reticulata – 125, 150 and 200 mg L^?1, S. discus – 75 and 100 mg L^?1 and X. helleri – 125 and 150 mg L^?1. The induction time generally decreased significantly with increasing concentration of MS‐222 for all of the species evaluated. The recovery time had a tendency to increase with the increase of the MS‐222 concentration for D. rerio, P. reticulata and S. discus. On the other hand, X. helleri recovery time decreased with the increase of MS‐222 concentration. MS‐222 proved to be effective in anaesthesia for all the freshwater ornamental species studied. The main results clearly show that the optimal dose to anesthetize is fish species dependent and it is completely wrong to extrapolate optimal anaesthetic concentrations between different species.     

The efficacy of Aqui‐S vet. (iso‐eugenol) and metomidate as anaesthetics in European eel (Anguilla anguilla L.), and their effects on animal welfare and primary and secondary stress responses

The effects of 40 mg/L metomidate and 300 mg/L iso‐eugenol (i.e. 600 mg/L Aqui‐S vet.) anaesthesia on the stress response in European silver eel during a process of complete anaesthesia, recovery and long‐term survival were studied. Metomidate is a hypnotic agent with no analgesia, whereas Aqui‐S vet. is a true anaesthetic. There was no difference between fish exposed to Aqui‐S vet. and metomidate, with respect to time to anaesthesia (mean 3.8 and 2.6 min respectively) and recovery (mean 7.6 and 6.5 min respectively). For the metomidate group, the plasma cortisol concentration increased to a peak during the recovery phase, and was significantly higher than for the Aqui‐S vet. group, which showed no such increase. Fish were kept in tanks with seawater for monitoring of long‐term survival (4 months) after anaesthesia treatment, and no mortality was observed in either group. The results indicate that metomidate is a potential stressor to European eels during exposure, and we do not recommend using metomidate in this species. Aqui‐S vet., however, seems promising as a stress‐reducing anaesthetic for European eel, and if used properly could improve animal welfare and survivability during and after common ecology‐related procedures, as capture, tagging and size measuring.     

Influence of temperature and size on menthol anaesthesia in Chinese grass shrimp Palaemonetes sinensis (Sollaud, 1911)

Chinese grass shrimp, Palaemonetes sinensis (Sollaud, 1911), is an economically important freshwater shrimp in China and adjacent areas. It is advisable to use anaesthesia in this species for certain handling and shipping operations; however, there have been no investigations into the recommended dosages. Here, the influence of five menthol concentrations (varying from 100 to 500 mg/L) on three different size classes of P. sinensis were examined at 8, 12, 16, 20, 24 and 28°C. Induction and recovery times for each shrimp were recorded, and effects of temperature, size and menthol dose on induction and recovery times were observed. Results showed that menthol dose, water temperature and shrimp size significantly influence anaesthesia in P. sinensis. Induction time decreased linearly with increasing water temperature and concentration of menthol, and increased with body weight. However, recovery times lengthened with concentration and temperature, and became shorter with body weight. Average body weight of the shrimps generally decreased after anaesthesia. Mortality of shrimps was correlated with temperature, dose and size. These results suggest that menthol is an effective rapid anaesthetic for P. sinensis, but there may be some disadvantages, including slow recovery and possible mortality for small shrimps and at higher temperatures and dosages.     

Menthol and 1,8‐cineole as new anaesthetics in common carp,Cyprinus carpio (Linnaeus, 1758)

This study describes anaesthetic efficacy of menthol and 1,8‐cineole in common carp, Cyprinus carpio, in comparison with eugenol. Common carp fingerlings were exposed to eugenol: 5, 10, 15, 25, 35, 50, 75, 100, 150 and 200 ppm; menthol: 5, 10, 15, 25, 50, 100, 200, 300, 400, 500 and 600 ppm; 1,8‐cineole: 100, 150, 200, 300, 400, 500, 600, 700 and 800 ppm. Induction time and recovery time were recorded. Results showed that menthol and 1,8‐cineole anesthetized the fish at higher concentrations compared to eugenol. The fish exposed to menthol showed common fish behavioural responses to anaesthesia, similar to eugenol. But, 1,8‐cineole‐exposed fish showed tail‐up swimming, which was not observed before. Also, 1,8‐cineole failed to completely cease muscle tone. Exposure to 200 ppm eugenol and 600 ppm menthol resulted in 40% and 20% mortality, respectively. Induction time was exponentially dependent on anesthetic concentrations. Recovery time was linearly correlated to eugenol and menthol, but not 1,8‐cineole concentrations. Recovery time was exponentially dependent on induction time in the fish anesthetized with eugenol and menthol, but not 1,8‐cineole. Menthol and 1,8‐cineole are recommended for carp anaesthesia. Menthol is capable to anesthetize common carp within 1–3 min at 118–512 ppm. Common carp anesthetized with 108–133 ppm menthol recovers within 5 min. 1,8‐cineole failed to anesthetize common carp within less than 150 s at 300–800 ppm concentrations. However, it anesthetizes carp within 3 min at 595 ppm concentration. Also, 1,8‐cineole is not recommended for fish surgery. Both menthol and 1,8‐cineole were less efficacious than eugenol.     

Anaesthetic efficacy and biochemical effects of 1,8‐cineole in rainbow trout (Oncorhynchus mykiss,Walbaum, 1792)

The aim of the present study was to investigate the anaesthetic efficacy and biochemical effects of 1,8‐cineole (cineole) in rainbow trout (Oncorhynchus mykiss). The fish were exposed to 200, 300, 400, 500, 600 and 800 μl/L cineole and time of induction of anaesthesia and recovery from anaesthesia were recorded. Thereafter, the fish haematological and biochemical responses to anaesthesia with different concentrations of cineole were studies. Moreover, the haematological and biochemical response of fish anaesthetized for 300 s with either cineole (283 μl/L) or eugenol (25 μl/L) were compared. Cineole at the concentrations of 200–800 μl/L induced stages 2, 3 and 4 anaesthesia within 109‐29.3, 226‐59 and 418‐117 s respectively. Increase in anaesthesia induction time led to higher stress responses and enzymes’ activity characterized by elevation in red blood cell (RBC), white blood cell (WBC), blood haematocrit and haemoglobin, and plasma cortisol, glucose and lactate levels, and aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase and creatine phosphokinase (CPK) activities. However, cineole concentrations had no significant effects on plasma alanine aminotransferase (ALT) activity and ions levels. Comparison of cineole with eugenol showed that cineole showed less change in blood RBC and plasma AST, ALP, CPK, cortisol and glucose levels compared to eugenol. In conclusion, cineole is efficient to anaesthetize trout at concentrations of 200–800 μl/L. Increase in cineole concentration shortens anaesthesia induction time, stress response and probably tissue damage. The concentrations of 600–800 μl/L cineol is recommended for rapid sampling as it causes the least stress and enzymatic responses. The present results suggest that cineole causes slightly lower side effects in trout compared to eugenol.     

The efficacy of clove oil as an anaesthetic for rainbow trout, Oncorhynchus mykiss (Walbaum)

The anaesthetic effects of clove-oil-derived eugenol were studied in juvenile rainbow trout, Oncorhynchus mykiss (Walbaum). Acute lethality and the effects of multiple exposures to eugenol were measured. The estimated 8-96 h LC₅₀ for eugenol was found to be approximately 9 p.p.m. Times to induction and recovery from anaesthesia were measured and compared with MS-222 under similar conditions. Eugenol generally induced anaesthesia faster and at lower concentrations than MS-222. The recovery times for fish exposed to eugenol were six to 10 times longer than in those exposed to similar concentrations of MS-222. Clove oil eugenol was determined to be an acceptable anaesthetic with potential for use in aquaculture and aquatic research. Doses of 40-60 p.p.m. eugenol were found to induce rapid anaesthesia with a relatively short time for recovery in juvenile trout.     

Clove oil attenuates stress responses in lambari,Astyanax altiparanae

The effects of clove oil anaesthetic on mitigating the physiological responses to air exposure, a stressful and routine situation in fish farming, laboratory conditions and sport fishing (catch and release), were evaluated in lambari (A. altiparanae). Adult females (n = 80) were randomly sorted to receive one of four treatments: control, anaesthesia (clove oil 50 mg/L), stress (5 min air exposure) and pre‐anaesthesia associated to stress. Their cortisol, glucose, lactate and haematocrit levels, the hepatosomatic index (HSI), liver and muscle glycogen, lipid peroxidation and the enzymatic activity of lactate dehydrogenase (LDH), catalase (CAT) and glutathione reductase (GR), were recorded. Glucose levels increased (53.9%) after anaesthesia and/or stress. The stress situation increased plasma cortisol (146.6%), lactate (294.6%) and lipid peroxidation in white muscle (45%) and decreased glycogen in white muscle (40.1%). The haematocrit increased after stress or anaesthesia (7.9%) while the liver glycogen and HSI did not change. Anaesthesia or stress did not affect the LDH but reduced the activity of CAT (46.1%) and GR (30.3%). We concluded that the anaesthetic clove oil in the concentration 50 mg/L modulates the physiological responses to air exposure stress improving the welfare; air exposure and clove oil affect the antioxidant defences of lambari; the activity of CAT and GR and the concentration of MDA can be used as biomarkers of stress in A. altiparanae.     

Anaesthesia of Atlantic cod (Gadus morhua) — Effect of pre-anaesthetic sedation, and importance of body weight, temperature and stress

The efficacy of the anaesthetic agents benzocaine, metacaine (MS-222), metomidate and 2-phenoxyethanol was studied in Atlantic cod (Gadus morhua) with average body weights of 10 ± 4 g, 99 ± 33 g and 1022 ± 274 g at water temperatures of 8 °C and 16 °C. The agents were tested individually and as combination anaesthesia comprising pre-anaesthetic sedation with a low dosage of metomidate or 2-phenoxyethanol followed by anaesthesia with benzocaine or MS-222. All agents were administered through bath immersion with an exposure time of 5 min. The different treatments resulted in average induction and recovery times ranging from 52 ± 6 s to 182 ± 16 s and 77 ± 26 s to 659 ± 46 s respectively. Induction and recovery times varied in relation to water temperature and were generally shorter at 16 °C for all weight groups and treatments compared to 8 °C. For benzocaine and MS-222 induction and recovery times were found to increase with increasing body weight. For metomidate the recovery time increased with increasing weight whereas there were no weight related differences in induction time. No differences in either induction or recovery times associated to body weight were found for 2-phenoxyethanol. Acute stress prior to anaesthesia with MS-222 resulted in significantly shorter induction time and prolonged recovery time, as well as deeper anaesthetised fish. The dosage of MS-222 had to be reduced in order to avoid mortality in fish subjected to acute stress. Combination anaesthesia allowed a reduction of the dosages used for inducing anaesthesia and produced markedly reduced recovery times compared to agents administered individually.     

On the efficacy and mode of action of 2-phenoxyethanol as an anaesthetic for goldfish, Carassius auratus (L.), at different temperatures and concentrations

The objective of this study was to assess the efficacy and mode of action of 2-phenoxyethanol as an anaesthetic for two size classes of goldfish, Carassius auratus (L.), at three different temperatures. Goldfish (2.15 ± 0.05 g, and 9.19 ± 0.17 g) were exposed to 0.3, 0.4 and 0.5 ml 2-phenoxyethanol l¹ at 20, 25, and 30^oC. Time needed to induce anaesthesia was dependent on concentration and water temperature. At temperatures at and below 25^oC, 0.4 ml l¹ was needed to induce total loss of equilibrium within less than 15 min. Above 25^oC, 0.5 ml l¹ was required to induce anaesthesia. Recovery rate was independent of the length of anaesthesia, which indicates that the anaesthetic is taken up and lost via a concentration gradient at the gill membrane and skin/solution interface. Fish recovered within less than 10 min after they had been taken out of the anaesthetic solution. In a second experiment, goldfish responded to a repeated exposure to 2-phenoxyethanol daily over a period of 14 days with increased tolerance, which indicates a habituation response to the anaesthetic. The use of 2-phenoxyethanol as an anaesthetic both for short-term anaesthesia and for anaesthesia under transport conditions is discussed.     

Hot water immersion lowers survival,shell growth rate and lysosomal membrane stability of oysters Crassostrea virginica (Gmelin)

This study compares the effect of two anti‐fouling treatments, hot water immersion (15 s at 60°C) and air drying (72 h) on the physiological status of the Eastern oyster Crassostrea virginica. The negative impact of hot water immersion was greater than that of air drying, but varied depending on the initial size of the oysters (40 vs. 60 mm shell height) and the time of the year (June vs. August). Groups treated with hot water exhibited a higher proportion of haemocytes with destabilized lysosomal membranes (HDLM; 47.5 ± 3.1%) than those exposed to air drying (37.5 ± 2.9%). This suggests that the oyster immunocompetency may be lowered by hot water immersion. Overall, the large oysters had lower HDLM values (32.9 ± 3.5%) than the small individuals in June (45.7 ± 2.8%) but similar values in August (46.6 ± 3.5%). Small oysters subjected to hot water immersion in June exhibited a 50% reduction in shell growth and a 50% mortality rate after one month. Our results indicate that air drying is more suitable than hot water immersion as an anti‐fouling treatment for <45 mm oysters.     

Effects of adding sucrose on Penaeus monodon (Fabricius, 1798) growth performance and water quality in a biofloc system

A 60‐day indoor growth trial was conducted to study the effects of biofloc on the growth performance of a Penaeus monodon (Fabricius, 1798), water quality and biological indicators including biofloc volume, chlorophyll‐a, heterotrophic bacteria and Bacillus quantity. Two concentrations of sucrose (0 and 75%) were added daily to the P. monodon culture systems (2.94 ± 0.11 g), which were conducted indoors in fibre‐glass tanks (500 L). Results showed that the final body weight and weight gain of the adding 75% sucrose group were significantly higher (P < 0.05) than that of the control, as well as significantly (P < 0.05) improved specific growth rates and survival rates, and reduced feed coefficient. Adding 75% sucrose promoted heterotrophic bacteria, Bacillus and phytoplankton reproduction, and significantly (P < 0.05) reduced the concentration of ammonia‐N (NH₄‐N), nitrite‐N (NO₂‐N) and nitrate‐N (NO₃‐N). The changes of water quality indicators in the two groups showed the similar trend at the end of the experiment, and the ammonia‐N, nitrite‐N, nitrate‐N and phosphate‐P concentrations in the 75% sucrose group were significantly (P < 0.05) lower than those of the control group, Chlorophyll‐a concentrations peaked at 389.12 μg/L in the biofloc sucrose group at 18:00 h, and heterotrophic bacteria peaked 8 h after sucrose was added. The addition of sucrose also reduced the pH of the water. Our research showed that adding sucrose promoted biofloc formation and shortened the formation time; increased the number of heterotrophic bacteria and algae which might play a role in improving water quality by assimilating ammonia‐N and other harmful substances in the water; supplemented food for P monodon growth; and reduced the feed coefficient.     

Water parameters affect anaesthesia induced by eugenol in silver catfish,Rhamdia quelen

The present study investigated the effects of water pH (5.0, 7.0 and 9.0), hardness (0, 20 and 120 mg CaCO₃ L^?1) and temperature (15, 23 and 30 °C) on the induction of sedation and anaesthesia, and subsequent recovery, of silver catfish exposed to eugenol. Moreover, the blood gas tensions (PvO₂ and PvCO₂) and blood pH in silver catfish acclimated to these temperatures were investigated after exposure to eugenol. Water pH, hardness, temperature and fish size affect the efficacy of eugenol in silver catfish, particularly at the lower concentrations tested (20 and 30 mg L^?1). Sedation of this species can be induced at concentrations as low as 20 mg L^?1, but for anaesthesia, a concentration of at least 40 mg L^?1 of eugenol must be used to compensate for the influence of fish size and water quality. Blood gas tension and pH were affected by eugenol anaesthesia, but only in fish acclimated to 30 °C.