The ability of clove oil and MS-222 to minimize handling stress in rainbow trout (Oncorhynchus mykiss Walbaum)

The effects of 60‐mg L^?1 clove oil and 60‐mg L^?1 tricaine methanesulphonate (MS‐222) on the blood chemistry of rainbow trout were compared after exposure to handling stress via caudal puncture blood sampling. Fish sampled by caudal puncture and subsequently exposed to anaesthetics showed a typical handling stress response over a 48‐h period. There were no significant differences between the responses of fish exposed to equal concentrations of clove oil and MS‐222, with the following exceptions: the blood glucose at full anaesthesia, and lactate at full recovery increased significantly in the clove oil‐exposed fish. In a subsequent experiment, the stress response observed in fish sampled by caudal puncture and exposed to clove oil and MS‐222 was compared with a non‐anaesthetized control group. The increases in plasma cortisol levels were significantly lower at recovery in fish treated with either anaesthetic compared with the control fish. Fish exposed to MS‐222 had significantly higher cortisol levels at 1 h. These findings show that few differences exist between the anaesthetic effects of clove oil and MS‐222 on the physiological response of fish to stress. However, clove oil is more effective at reducing the short‐term stress response induced by handling and blood sampling, and is recommended as an alternative fish anaesthetic.     

Olfactory nerve response of masu salmon (Oncorhynchus masou Brevoort) and rainbow trout (O. mykiss Walbaum) to clove oil and MS-222

Two anaesthetics, clove oil and methane sulphonate (MS-222), were examined for their effects on the olfactory nerve response of masu salmon ( Oncorhynchus masou Brevoort) and rainbow trout ( O. mykiss Walbaum). Exposing both species to clove oil for 3 min at concentrations of 50 and 100 mg L⁻¹, or for 10 min at 50 mg L⁻¹, did not significantly reduce their olfactory response. Directly applying clove oil anaesthesia to the olfactory epithelium significantly reduced olfactory response though after 20 min, olfactory response recovered to 70% and 52% of pre-treatment levels in masu salmon and rainbow trout respectively. Compared with the post-anaesthetic recovery of responses after clove oil (50 mg L⁻¹), buffered MS-222 (100 mg L⁻¹) with NaHCO₃ (100 mg L⁻¹), and unbuffered MS-222 (100 mg L⁻¹) treatment for 3 min, the response after MS-222 treatment declined gradually and significantly, but not after clove oil and MS-222+NaHCO₃ treatments. Clove oil appears to be an effective and relatively safe anaesthetic for salmonids with little long-term impact on their olfactory response, which plays a crucial role in their life history.     

MS-222和丁香油对黄条鰤麻醉效果研究

黄条鰤(Seriola aureovittata)是一种全球水域广泛分布的具有长距离洄游特性的大型大洋性鱼类,其生性凶猛、游泳速度快,人工养殖条件下对环境刺激的应激反应强烈。为开发适宜黄条鰤实验和生产操作的低应激处理技术,采用实验生态学、生理学手段,首次研究了2种温度条件下(20℃和24℃)、2种麻醉剂(MS-222和丁香油)对1龄黄条鰤的麻醉效果,通过行为判别与生理指标分析(皮质醇、肾上腺素和葡萄糖的水平),确定了2种麻醉剂对黄条 的最适麻醉时间、复苏时间和麻醉剂量。根据黄条鰤麻醉和复苏时的行为特征,将麻醉过程分为7个时期,复苏过程分为6个时期。在水温为20℃和24℃时,MS-222麻醉黄条 的最佳浓度分别为100~120 和100 mg/L,而丁香油的最佳浓度均为40 mg/L。在此条件下,实验鱼均可在3 min之内入麻,5 min之内复苏。随着麻醉剂浓度的增加,黄条鰤入麻时间呈缩短趋势,复苏时间呈延长趋势。水温的升高降低了入麻时间,而对复苏时间无明显影响。在水温20℃和24℃条件下,40 mg/L的丁香油麻醉黄条鰤后,血清皮质醇水平分别于6 h和12 h达峰值;在水温24℃条件下,100 mg/L的MS-222麻醉黄条鰤后,血清肾上腺素水平于12 h达峰值,其他各实验组血清肾上腺素、葡萄糖和皮质醇水平均在24 h达峰值。2种温度条件下,实验鱼血清中3种应激相关生理指标在复苏72 h后均显著降低至初始水平以下。本研究结果可为制定规范化的黄条鰤实验与养殖生产操作技术提供参考依据。     

Cholesterolaemia and triacylglycerolaemia in farmed rainbow trout,Oncorhynchus mykiss

Physiological (reference) value intervals determined by the lower 2.5% and upper 97.5% quantiles were calculated for blood plasma cholesterol (CHOL) and triacylglycerol (TGL) in farmed rainbow trout, Oncorhynchus mykiss, in raceway culture. The fish were given dry pelleted diets that contained 37–47% crude protein and 7–14% crude fat and were kept at a stocking density of 50 kg m^?3 in tanks provided with running freshwater at an ambient temperature of 3–16°C and dissolved oxygen, 8.4–11.5 mg L^?1. Blood was sampled between September and November at a photoperiod of 9:00–13:00 hours: 11:00–15:00 hours (light:dark). Cholesterol levels were significantly (P = 0.0001) greater in males (4.7–12.1 mmol L^?1, n = 34, mean weight 406 ± 138 g) than immature females (3.2–9.7 mmol L^?1, n = 386, mean weight 416 ± 147 g). Physiological range for TGL in immature females and males was 2.4–14.4 mmol L^?1 (n = 249, mean weight 418 ± 149 g). The distribution and density of the quantiles in the tested reference group were made possible by the use of histograms, which showed normal distributions for CHOL in males and in females, but not for TGL, in which a sinistral asymmetry was found. Correlation and regression analyses indicated significant (P = 0.0000) dependence between the CHOL and total protein (r² = 76.2%), CHOL and Fulton's condition factor (r² = 43.3%) and CHOL and absolute weight of liver (r² = 45.5%). Fluctuation in cholesterolaemia and triacylglycerolaemia, depending on nutrition and the aquaculture method is discussed below.     

Effects of grape Vitis vinifera seed oil supplementation on growth,survival, fatty acid profiles,antioxidant contents and blood parameters in rainbow trout Oncorhynchus mykiss

In this study, effects of grape Vitis vinifera seed oil supplementation (0, 250 mg, 500 mg, 1,000 mg/kg feed) for 60 days on rainbow trout Oncorhynchus mykiss juveniles were investigated. The average final fish weights were not different between the control group and 500 mg/kg feed group (p > .05), but 250 mg/kg feed group were lower than those of the control group, and 1,000 mg/kg feed group were higher than those of the control group. Although the SFA, MUFA and n3 ratios of the control group were higher than that of the other groups, no statistical difference was found. The highest fatty acids in the fish were palmitic acid C 16: 0 in the SFA group and C 18: 1n‐9 and docosahexaenoic acid C 22: 6n‐3 in the PUFA group. The EPA ratio was significantly increased in all groups compared with the control group. The DHA ratio was significantly higher in the control group than in the experimental groups, and there was no difference between the experimental groups. Urea were significantly lower in the 250 mg/kg feed group. Total protein and aspartate aminotransferase were significantly increased in the 1,000 mg/kg feed group. Only the 1,000 mg/kg feed group showed an increase in antioxidant activities, and this concentration also yielded positive results in terms of growth performance, survival rates, fatty acid composition and antioxidant enzyme activities in rainbow trout juvenile and can be used as a natural feed additive.     

Effects of anaesthesia with MS-222, clove oil and CO2 on feed intake and plasma cortisol in steelhead trout (Oncorhynchus mykiss)

We examined the effects tricaine methanesulfonate (MS-222), clove oil and CO₂ on feed intake and cortisol response in steelhead trout, Oncorhynchus mykiss. Even though a body of literature exists about the effects of different anaesthetics on fish, no comparative information seems to be available about their effects on feed intake after anaesthesia, which would be important to know especially in aquaculture research. We anaesthetised juvenile steelhead trout with these three anaesthetics, and then sampled them 4, 24 and 48 h later. Fish in all groups ate relatively well already 4 h after anaesthesia. However, feed intake in fish treated with clove oil or MS-222 was lower than in the controls. There were no differences in feed intake among anaesthetised groups. Plasma cortisol concentrations were elevated 48 h after anaesthetisation, but the treatment means were equal throughout the experiment. Our results support previous findings that clove oil is a reasonable alternative to MS-222.     

Red blood cell indices of rainbow trout Oncorhynchus mykiss (Walbaum) in aquaculture

A direct non‐parametric method was used to calculate reference (physiological) haematology values for farmed 10–12‐month rainbow trout of the Kamloops strain (mean weight: 330±131 g) with respect to red blood cell counts (RBCc), haematocrit values (Hct), haemoglobin concentrations (Hb), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentrations (MCHC). The fish in the selected reference group (n=798) were given dry pelleted diets that contained 37–47% crude protein, 7–18% crude fat and 108–300‐mg vitamin E, 1.08–5‐mg folic acid, 0.018–0.05‐mg vitamin B₁₂, 48–64.5‐mg iron, 4.5–8.4‐mg copper and 0.18–0.24‐mg selenium supplied per kg of diet. Ethoxyquin and butylhydroxytoluol were used to protect the fat component against oxidation. The fish were kept at a stocking density of 50kg per cubic metre in tanks provided with running freshwater (dissolved oxygen 8.4–11.5 mg L^?1, with O₂ saturation of 77–98%) at an ambient temperature of 0.2–16°C. Blood was sampled between September and November at a photoperiod of 9–13 h:11–15 h (light:dark). Reference ranges for the preceding haematological indices were as follows in immature females (males): RBCc, 0.77–1.42T L^?1 (T – tera, 10¹²) (0.98–1.55T L^?1); Hct, 0.304–0.502 (0.34–0.546); Hb, 54–93 g L^?1 (59–97 g L^?1); MCV, 282–469 fL (279–434 fL); MCH, 51–86 pg (47–78 pg); MCHC, 0.15–0.22 (0.15–0.2). In males, values for RBCc, Hct and Hb were significantly higher (P=0.01 and 0.0000 respectively) and those for MCV, MCH and MCHC were significantly lower (P=0.01 and 0.0002 respectively) than in immature females. Nutritional and environmental factors affecting erythropoiesis in trout and some correlations between haematological (RBCc, Hb, Hct) and biochemical indices of the blood plasma (total protein, cholesterol) are discussed.     

Total calcium and inorganic phosphate in the blood plasma of farmed rainbow trout, Oncorhynchus mykiss

Reference (physiological) value intervals determined by the lower 2.5% and upper 97.5% quantiles were calculated for total calcium (Ca _t ) and inorganic phosphate (P) in farmed rainbow trout, Oncorhynchus mykiss , at an age of 10–12 months in raceway culture. Total calcium levels were significantly ( P ≤0.0009) greater in males (2.63–4 mmol L⁻¹, n =47, mean weight 359±142 g) than in immature females (2.48–3.97 mmol L⁻¹, n =410, mean weight 400±145 g). The reference range for P in immature females and males was 3.09–9.32 mmol L⁻¹ ( n =467, mean weight 401±144 g). The distribution and density of the quantiles in the tested reference group were made possible by the use of histograms, which showed normal distributions for Ca _t in males but not for Ca _t and P in immature females and males, in which a sinistral asymmetry was found.     

MS-222、丁香酚对多鳞四指马（鲅）幼鱼麻醉效果的研究

本文研究了MS-222和丁香酚两种麻醉剂在不同浓度下对多鳞四指马（鲅）幼鱼的麻醉效果,观测了其不同麻醉阶段的表现,探讨了麻醉时间和温度对其入麻和复苏的影响.结果表明 MS-222、丁香酚分别在55 mg/L、10 mg/L以上才对幼鱼有麻醉效果,最适麻醉剂浓度分别为20±5 mg/L和80±10 mg/L;随着浓度的上升,入麻时间缩短,复苏时间增加.随着温度的上升,入麻时间缩短,复苏时间增加.     

A study of gross,histological and blood biochemical changes in rainbow trout,Oncorhynchus mykiss (Walbaum), with rainbow trout gastroenteritis (RTGE)

The mechanisms behind the pathogenesis of rainbow trout gastroenteritis (RTGE) are still unknown. This study examined the macroscopic and microscopic changes in trout with RTGE (RTGE+), as well as the blood chemistry. A total of 464 rainbow trout were sampled from 11 sites in the UK, comprising 152 RTGE+ fish and 330 random, apparently healthy fish. A case definition for RTGE was assessed by the analysis of its agreement with three laboratory tests: histopathology, packed cell volume and kidney bacteriology. Cluster analysis indicated the presence of three distinct presentations within the population of RTGE+ fish. Cluster A included gross signs associated with moribund RTGE+ fish, and clusters B and C identified gross signs consistent with concurrent diseases, notably furunculosis, enteric redmouth and proliferative kidney disease. The information gained was used to select RTGE+ fish without concurrent disease for the analysis of RTGE pathogenesis with blood biochemistry. This analysis revealed a severe osmotic imbalance and a reduced albumin/globulin ratio as indicatives of selective loss of albumin. These findings are compatible with a protein losing enteropathy.     

MS-222对细鳞鲑和美洲红点鲑的麻醉效果研究

通过测定不同质量浓度MS-222对细鳞鲑和美洲红点鲑的麻醉效果,检测麻醉时鱼的呼吸频率以及空气暴露时长对鱼体复苏的影响,探究两种鱼类在渔业生产和科研实验中的最适麻醉剂浓度。结果显示：当MS-222质量浓度为80、100 mg/L时,细鳞鲑麻醉所需的时间显著少于40、60 mg/L浓度组,而显著多于其余浓度组(P<0.05)。当MS-222质量浓度为100 mg/L时,细鳞鲑的复苏时长最短(P<0.05)。当MS-222质量浓度为80 mg/L时,美洲红点鲑麻醉所需的时间显著少于40、60 mg/L浓度组,而显著多于其余浓度组(P<0.05)。当MS-222质量浓度为100 mg/L时,美洲红点鲑的复苏时长显著大于40、60和80 mg/L浓度组,而显著小于其余浓度组(P<0.05)。比较MS-222对两种鱼的麻醉效果,美洲红点鲑对MS-222更为敏感。两种鱼的呼吸频率总体上都随着麻醉程度的加深而下降,当空气暴露时长少于5 min时,细鳞鲑的复苏时长先减少后增加,美洲红点鲑的复苏时长则呈逐渐减少趋势。     

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.     

Relationships between growth and disease resistance in rainbow trout,Oncorhynchus mykiss (Walbaum)

Rainbow trout from 23 families were evaluated for growth and resistance to the bacterial coldwater disease (BCWD) caused by Flavobacterium psychrophilum and infectious haematopoietic necrosis (IHN) caused by IHN virus. Average family weights were between 161 and 263 g with an average of 225 g at 213 days post‐fertilization with specific growth rates ranging from 2.37 to 2.88. Per cent survival of fish challenged with F. psychrophilum was between 18% and 100%, while for those challenged with IHNV, the range was between 12% and 93%. Significant positive correlations were found for end body weight and resistance to IHN (P < 0.05) and for early body weight and resistance to BCWD (P < 0.1). However, no significant correlations were detected between resistance to both pathogens or disease resistance and overall genetic diversity or diversity within the major histocompatibility locus.     

Use of probiotics to control furunculosis in rainbow trout, Oncorhynchus mykiss (Walbaum)

Aerobic heterotrophic bacteria were isolated from the intestinal contents of Atlantic salmon, Salmo salar , rainbow trout, Oncorhynchus mykiss , and turbot, Scophthalmus maximus , on tryptone soya agar and De Man Rogosa and Sharpe agar, of which 11 of 177 (6% of the total) of the isolates were antagonistic to Aeromonas salmonicida . Four of these cultures, which were identified tentatively as A. hydrophila , Vibrio fluvialis , Carnobacterium sp. and an unidentified Gram-positive coccus, were beneficial to fish when fed singly or as an equi-mixture. Feed supplemented with the putative probiotics indicated survival of the organisms in the gastrointestinal tract for 7 days. Feeding with the probiotics for 7 and 14 days led to better survival following challenge with A. salmonicida . There was no indication of serum or mucus antibodies to A. salmonicida , but there was an increased number of erythrocytes, macrophages, lymphocytes and leucocytes, and enhanced lysozyme activity in the fish.     

Competitive exclusion of brown trout Salmo trutta L., by rainbow trout Oncorhynchus mykiss Walbaum, in lake tributaries, New Zealand

Spawning runs of brown trout Salmo trutta L., and rainbow trout, Oncorhynchus mykiss Walbaum, were compared in three tributaries of two lakes for timing, numbers and redd characteristics. Redd characteristics for the two species in four other streams were examined. No significant differences in redd characteristics between the two species were observed, but in all lake tributaries the possibilities of superimposition of spring spawning rainbow trout on redds of brown trout were examined. A common feature was the high ratio of numbers of spawners to the area of preferred spawning gravel, and it was concluded that in the lake tributaries, superimposition resulted in increased mortality of brown trout eggs, and in one case elimination of the brown trout population. Management possibilities for control of the species proportions in comparable situations are discussed.     

Efficacy of clove oil and ethanol against Saprolegnia sp. and usability as antifungal agents during incubation of rainbow trout Oncorhynchus mykiss (Walbaum) eggs

Inhibitory concentrations of clove oil and ethanol against growth of Saprolegnia sp. hyphae were screened by a modification of the hemp (Cannabis sativa L.) seed MicroPlate (HeMP) method and their usability as antifungal agents during incubation of rainbow trout Oncorhynchus mykiss eggs was tested. In vitro experiment showed that in continuous static exposure, clove oil at 100 mg L^?1 significantly inhibited the growth of Saprolegnia, whereas in bath exposures, clove oil at 500 mg L^?1 had no significant effect at any exposure time tested (15, 60 and 240 min), but clove oil at 10 000 mg L^?1 significantly inhibited growth at all exposure times. Clove oil and ethanol treatments had no visible effects on the onset or spread of the fungus during incubation of rainbow trout eggs. Clove oil at 1000 mg L^?1 resulted in 95–100% mortality before the eyed stage was reached. Sublethal concentrations of clove oil and ethanol had no effects on the development rate of the embryo or growth and yolk utilization efficiency after hatching. This study suggests that clove oil and ethanol may not be options in controlling aquatic fungi infestations during incubation of rainbow trout eggs.     

MS-222对中华鲟和施氏鲟的麻醉试验

以20~100 mg/L的MS-222(Tricaine M ethanesu lphonate)在水温15~25℃条件下对中华鲟(Acipensersinensis)和施氏鲟(A.schrenckii)的麻醉效果进行试验,中华鲟或施氏鲟分别达到不同程度的麻醉状态甚至出现死亡。30~40 mg/L的MS-222具有较好的麻醉效果,鱼的活动量明显下降,可保持镇静并维持身体平衡和正常体位,麻醉48 h后,在清水中数分钟内可恢复正常游动状态,适合于长途运输。50~100 mg/L的MS-222对中华鲟和施氏鲟具有较强的麻醉作用,适合于进行短时间的操作,如捉拿、称重、测量、外科手术、体外标记等。中华鲟和施氏鲟在高剂量MS-222的作用下,在进入生物学死亡阶段前,存在一个临床死亡阶段,这个阶段长达1h左右,在这个阶段,对它们进行“人工呼吸”是行之有效的。