急性氨氮胁迫及毒后恢复对团头鲂幼鱼鳃、肝和肾组织结构的影响

为进一步了解氨氮对团头鲂幼鱼的毒性毒理影响,以体质量为(14.27±0.01)g的团头鲂幼鱼为研究对象,研究了氨氮胁迫对其鳃、肝、肾组织结构的影响。实验首先进行96h的氨氮胁迫,得出96h LC50,在此基础上,设置对照组(0.472 mg/L)和实验组(25 mg/L)两个氨氮浓度处理组,进行0、6、12、24、48h的氨氮胁迫,取样后剩余团头鲂幼鱼移入曝气自来水进行96h的毒后恢复实验。结果表明：96h LC50为56.492 mg/L;三种组织观察表明,氨氮胁迫6h,鳃丝毛细血管扩张,上皮组织增生;肝细胞肿胀,细胞核肿大,肝细胞空泡化;肾小球萎缩,肾小囊腔膨大,肾小管管腔缩小;胁迫12h,泌氯细胞增生,呼吸上皮细胞出现部分脱落;肝细胞水样变性、血窦扩张、细胞轮廓模糊,形成点状病灶;肾小管上皮细胞肿大、水样变性、浊肿;胁迫24h,鳃小片融合、变短,呼吸上皮细胞大面积脱落;肝细胞水样变性、血窦扩张严重,形成局部病灶;肾组织淋巴细胞浸润严重,充血,肾小球坏死,肾小管坏死;胁迫48h,鳃小片卷曲,上皮细胞部分脱落;肝细胞部分溶解、血窦扩张,形成点状病灶;肾小管上皮细胞坏死,肾小球坏死;96h恢复后,泌氯细胞和上皮组织增生严重;肝组织大面积细胞核肿大,血窦扩张;肾组织淋巴细胞浸润严重,肾小管坏死,肾小球坏死。实验表明,不同的器官之间病症的损伤程度是不同的,肝组织的损伤最严重,然后依次是鳃和肾。随着胁迫时间延长,鳃、肝和肾组织受到的损害增加,同时鱼体也产生防御反应,但96h的恢复期不足以让团头鲂幼鱼在胁迫中完全恢复,而恢复能力最差的是肾组织。     

Ammonia and nitrite toxicity to false clownfish <Emphasis Type="Italic">Amphiprion ocellaris</Emphasis>

False clownfish, Amphiprion ocellaris, is one of the most commercialized fish species in the world, highly produced to supply the aquarium market. The high stocking densities used to maximize fish production can increase ammonia and nitrite to toxic levels. In this study, A. ocellaris juveniles (1.20 ± 0.34 g) were exposed to six concentrations of ammonia ranged from 0.23 to 1.63 mg/L NH₃-N and eight concentrations of nitrite (26.3–202.2 mg/L NO₂ ^?-N). The LC₅₀- 24, LC₅₀-48, LC₅₀-72 and LC₅₀-96 h were estimated to be 1.06, 0.83, 0.75 and 0.75 mg/L for NH₃-N and 188.3, 151.01, 124.1 and 108.8 mg/L for NO₂ ^?-N. Analysis of gill lesions caused by sublethal concentrations of these nitrogenous compounds showed that both nitrogenous compounds induced tissue lesions such as hyperplasia of epithelium cells, hypertrophy of chloride cells and lamellar lifting to all concentrations tested. However, histopathological alterations were more conspicuous accordingly the increase of ammonia or nitrite in fish exposed to 0.57 mg/L NH₃-N or 100 mg/L NO₂ ^?-N. Based on our results, we recommend to avoid concentrations higher than 0.57 mg/L of NH₃-N and 25 mg/L of NO₂-N in water.     

Acute and chronic effects of aqueous ammonia on marbled spinefoot rabbitfish,Siganus rivulatus (Forsskål 1775)

Ammonia is a metabolite of aquatic organisms which might reach deleterious levels in intensive fish farms. The aim of the present study was to determine median lethal concentrations (96‐h LC₅₀) of total ammonia nitrogen (TA‐N) on marbled spinefoot rabbitfish (Siganus rivulatus) and chronic effects of TA‐N on survival, growth and behaviour of juvenile rabbitfish over a 50 day period. In the first experiment, fish were exposed to 0, 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 mg L^?1 TA‐N for 96 h and survival evaluated. In the second experiment, 12 fish were stocked per 50‐L tank and treated with one of 0, 2, 4, 6, 8, 10 and 12 mg L^?1 TA‐N with three replicate tanks per treatment. Survival and growth were determined and histopathological alterations of gills due to chronic ammonia exposure were studied by light and electron microscopy. The 96‐h LC₅₀ values were 16–18 mg L^?1 TA‐N. In the chronic exposure experiment, fish reared in water with 0 mg L^?1 TA‐N had 100% survival and had 50% weight increase in 50 days. Fish at 2 and 4 mg L^?1 TA‐N all died whilst fish in 6, 8, 10 and 12 mg L^?1 TA‐N survived and grew albeit less than in treatment 0 mg L^?1. Gills from ammonia treated fish displayed severe histological and ultrastructural alterations including hyperplasia, hypertrophy and fusion of secondary lamellae, aneurysms and presence of pleomorphic altered cells. Chronic exposure to ammonia is deleterious to marbled spinefoot rabbitfish and low concentrations of ammonia appear to kill the fish in <50 days whilst fish can survive for more than 50 days at concentrations between 6 and 12 mg L^?1 TA‐N.     

Acute toxicity of ammonia in Meagre (Argyrosomus regius Asso, 1801) at different temperatures

Argyrosomus regius (3.0 ± 0.9 g) were exposed to different concentrations of ammonia in a series of acute toxicity tests by the static renewal method at three temperature levels (18, 22 and 26°C) at a pH of 8.2. Low temperature clearly increased the tolerance of the fish to total ammonia nitrogen (TAN) and unionized ammonia (NH₃) (P < 0.05). While the 96‐h LC₅₀ values of TAN were 19.79, 10.39 and 5.06 mg L^?1, the 96‐h LC₅₀ of NH₃ were 1.00, 0.70 and 0.44 mg L^?1 at 18, 22 and 26°C respectively. The safe levels of NH₃ for A. regius was estimated to be 0.10, 0.07 and 0.04 mg L^?1 at 18, 22 and 26°C respectively (P < 0.05). This study clearly indicates that A. regius is more sensitive to ammonia than other marine fish species cultured on the Mediterranean and Eastern Atlantic coasts.     

Effect of feeding time on dietary protein utilization and growth of juvenile Senegalese sole (Solea senegalensis)

The present study aimed to determine the effect of feeding time on growth and nitrogen excretion in juvenile sole. An 84‐day growth trial was conducted, in which food was supplied to three triplicate groups of juvenile Senegalese sole (3 g wet weight) at different schedules – diurnal, nocturnal and mixed. At the end of the growth trial, ammonia and urea excretion was assessed during a 24 h cycle. Improved growth (1.3% vs. 0.9% day^?1, specific growth rate), higher nitrogen retention (0.35 vs. 0.27 g N kg^?1 day^?1), lower ammonia excretion (209 vs. 272 mg N‐NH₄ kg^?1 day^?1) and lower total nitrogen excretion (278 vs. 352 mg N kg^?1 day^?1) were found in daytime‐fed fish compared with night‐fed fish. Fish in the mixed feeding regime showed intermediate values of ammonia and total nitrogen excretion, but did not differ from day‐fed fish regarding the other parameters. Results indicate that juvenile sole at a period of their life cycle appear to use more efficiently dietary protein for somatic growth under a diurnal than under a nocturnal feeding regime. This suggest that at least during a time‐window in the juvenile rearing a diurnal feeding regime might be more effective in the production of this species.     

Acute toxicity of ammonia in Pacu fish (Piaractus mesopotamicus,Holmberg, 1887) at different temperatures levels

Piaractus mesopotamicus juveniles (total length 12 ± 0.5 mm) were exposed to different concentrations of ammonia‐N (un‐ionized plus ionized ammonia as nitrogen), using the static renewal method at different temperature levels (15, 20 and 25°C) at pH 7. The 24, 48, 72, 96 h LC₅₀ values of ammonia‐N in P. mesopotamicus juveniles were 5.32, 4.19, 3.79 and 2.85 mg L^?1 at 15°C; 4.81, 3.97, 3.25 and 2.50 mg L^?1 at 20°C; and 4.16, 3.79, 2.58 and 1.97 mg L^?1 at 25°C respectively. The 24, 48, 72, 96 h LC₅₀ values of NH₃‐N (un‐ionized ammonia as nitrogen) were 0.018, 0.014, 0.013, 0.009 mg L^?1 at 15°C temperature; 0.023, 0.019, 0.016 and 0.012 mg L^?1 at 20°C; 0.029, 0.026, 0.018 and 0.014 mg L^?1 at 25°C. The temperature increase from 15 to 25°C caused an increase of ammonia‐N susceptibility by 21.80%, 9.55%, 31.92% and 30.87%, after 24, 48, 72 and 96 h exposure respectively. Furthermore, we found that exposure of fish to ammonia‐N caused an elevation in total haemoglobin and blood glucose with an increase of 2 mg L^?1 concentration. Ammonia levels tolerated, especially in different temperatures levels, have important implications for the management of aquaculture.     

Acute ammonia toxicity and gill morphological changes of Japanese flounder Paralichthys olivaceus in normal versus supersaturated oxygen

Ammonia toxicity and morphological changes in gills of juvenile Japanese flounder Paralichthys olivaceus (5.76 ± 0.12 g) were investigated when fish were separately exposed to normal dissolved oxygen (DO) at 6.5 ± 0.5 mg L^?1 and supersaturated oxygen at 16.0 ± 2.0 mg L^?1 at different ammonia concentrations. Under normal oxygen, ammonia concentrations were tested from 0.04 (control) to 93.3 mg L^?1 total ammonia nitrogen (TAN), whereas under oxygen supersaturation, ammonia concentrations ranged from 0.04 (control) to 226.7 mg L^?1 TAN in the trial. After exposure to ammonia for 96 h, the ammonia LC₅₀ for fish was 62.48 mg L^?1 TAN (0.50 mg L^?1 NH₃–N) at normal oxygen and 160.71 mg L^?1 TAN (0.65 mg L^?1 NH₃–N) at oxygen supersaturation. Light microscopic observations confirmed that gill damage in normal oxygen was more profound than in oxygen supersaturation when fish were exposed to the same level of TAN (93.3 mg L^?1). Furthermore, electron microscopic scanning also showed more crimple, retraction and fibrosis on the secondary lamella surface in fish exposed to normal oxygen than those in fish exposed to supersaturated oxygen at the same TAN (93.3 mg L^?1). This study suggests that supersaturated oxygen can increase ammonia tolerance in Japanese flounder through reducing gill damage by ammonia, which partially explains the merit of using pure oxygen injection in intensive fish farming.     

Acute tolerance of juvenile Florida pompano, Trachinotus carolinus L., to ammonia and nitrite at various salinities

The acute tolerance of juvenile Florida pompano Trachinotus carolinus L. (mean weight±SE=8.1±0.5 g) to environmental unionized ammonia‐nitrogen (NH₃‐N) and nitrite‐nitrogen (NO₂‐N) at various salinities was determined via a series of static exposure trials. Median‐lethal concentrations (LC₅₀ values) of NH₃‐N and NO₂‐N at 24, 48, and 96 h of exposure were calculated at salinities of 6.3, 12.5 and 25.0 g L^?1 at 28 °C (pH=8.23–8.36). Tolerance of pompano to acute NH₃‐N exposure was not affected by salinity, with 24, 48 and 96 h LC₅₀ values ranging from 1.05 to 1.12, 1.00 to 1.08 and 0.95 to 1.01 mg NH₃‐N L^?1 respectively. Regarding NO₂‐N, tolerance of pompano to this environmental toxicant was compromised at reduced salinities. Median‐lethal concentrations of NO₂‐N to pompano at 24, 48 and 96 h of exposure ranged from 67.4 to 220.1, 56.9 to 140.7 and 16.7 to 34.2 mg NO₂‐N L^?1 respectively. The results of this study indicate that juvenile Florida pompano are relatively sensitive to acute NH₃‐N and NO₂‐N exposure, and in the case of the latter, especially at lower salinities.     

Ammonia tolerance of Litopenaeus vannamei (Boone) larvae

The tolerance of Litopenaeus vannamei larvae to increasing concentrations of total ammonia nitrogen (TAN) using a short‐term static renewal method at 26°C, 34 g L^?1 salinity and pH 8.5 was assessed. The median lethal concentration (24 h LC₅₀) for TAN in zoea (1‐2‐3), mysis (1‐2‐3) and postlarvae 1 were, respectively, 4.2‐9.9‐16.0; 19.0‐17.3‐17.5 and 13.2 mg L^?1TAN (0.6‐1.5‐2.4; 2.8‐2.5‐2.6 and 1.9 mg L^?1 NH₃‐N). The LC₅₀ values obtained in this study suggest that zoeal and post‐larval stages are more sensitive to 24 h ammonia exposure than the mysis stage of L. vannamei larvae. On the basis of the ammonia toxicity level (24 h LC₅₀) at zoea 1, we recommend that this level does not exceed 0.42 mg L^?1 TAN – equivalent to 0.06 mg L^?1 NH₃‐N – to reduce ammonia toxicity during the rearing of L. vannamei larvae.     

Influence of long term ammonia exposure on Atlantic salmon (Salmo salar L.) parr growth and welfare

The objective of this study was to determine the long‐term effects of ambient unionized ammonia nitrogen (NH₃‐N) combined with different feeding regimes on Atlantic salmon Salmo salar L parr growth, welfare and smoltification. Previous studies on the parr stage of Atlantic salmon have mostly focused on acute exposure, or at low temperatures. Atlantic salmon parr were exposed for 105 days (at 12°C, pH 6.8) to four sublethal ammonia concentrations ranging from 0.1 to 35 μg L^?1 NH₃‐N (0.1–25 mg L^?1 TAN) at two feeding levels: full feed strength (+20% overfeeding) and 1/3 of full feed strength. After 21 days, it was observed that 32 μg L^?1 NH₃‐N reduced growth rate of parr fed full ration, but this effect was not evident at the end of the exposure. Feed utilization was not affected by ammonia exposure at any sampling point. Increasing ammonia levels were associated with a higher prevalence and severity of gill damage at 22 days but not at the end of the exposure. The examination of welfare indicators revealed only a few pathologies, not related to ammonia exposure. In addition, higher ammonia concentrations did not appear to influence the development of hypo‐osmoregulatory ability during parr‐smolt transformation.     

An optimized and simplified method for analysing urea and ammonia in freshwater aquaculture systems

This study presents a simple urease method for analysis of ammonia and urea in freshwater aquaculture systems. Urea is hydrolysed into ammonia using urease followed by analysis of released ammonia using the salicylate‐hypochlorite method. The hydrolysis of urea is performed at room temperature and without addition of a buffer. A number of tests were performed on water samples obtained from a commercial rainbow trout farm to determine the optimal urease concentration and time for complete hydrolysis. One mL of water sample was spiked with 1.3 mL urea at three different concentrations: 50 μg L^?1, 100 μg L^?1 and 200 μg L^?1 urea‐N. In addition, five concentrations of urease were tested, ranging from 0.1 U mL^?1 to 4 U mL^?1. Samples were hydrolysed for various time periods ranging from 5 to 120 min. A urease concentration of 0.4 UmL^?1 and a hydrolysis period of 120 min gave the best results, with 99.6–101% recovery of urea‐N in samples spiked with 100 or 200 μg L^?1 urea‐N. The level of accurate quantification of ammonia using the method is 50 μg L^?1 NH₄⁺‐N, and the detection level is 5–10 μg L^?1 NH₄⁺‐N.     

Immune responses of Litopenaeus vannamei to non‐ionic ammonia stress: a comparative study on shrimps in freshwater and seawater conditions

To investigate the effect of non‐ionic ammonia (NH₃‐N) stress (0.1 and 0.5 mg L^?1) on the immunity of Litopenaeus vannamei cultured in long‐term freshwater, the total haemocyte count (THC), the activity of phenoloxidase (PO), nitric oxide synthase (NOS), superoxidase dismutase (SOD) and the content of malondialdehyde (MDA) were determined and further compared with those of seawater shrimps. The results showed that NH₃‐N stress significantly reduced THC and the activity of PO and SOD (P < 0.05). Under 0.1 mg L^?1 NH₃‐N stress, NOS activity increased first and then decreased significantly, while it dropped dramatically under 0.5 mg L^?1 NH₃‐N stress (P < 0.05). During NH₃‐N stress, MDA content increased continuously, and the MDA content in hepatopancreas of freshwater shrimps was higher than that of seawater shrimps. It was concluded that NH₃‐N stress significantly influenced the non‐specific immunity and could also upset the balance of antioxidant system of L. vannamei in both freshwater and seawater shrimps. Compared with in seawater, the shrimps in freshwater were more vulnerable to NH₃‐N stress because of higher lipid peroxidation and lower immunity.     

Tolerance of un‐ionized ammonia in live feed cultures of the calanoid copepod Acartia tonsa Dana

Optimal water quality is considered as being a restriction for marine copepod cultures for live feed. There is a lack of knowledge on the water‐quality conditions in copepod cultures and the effect on copepods. Few studies have investigated the effect of ammonia on copepods, and fewer reports No Observed Effect Concentrations (NOEC) and Lowest Observed Effect Concentrations (LOEC), which provides safety levels before cultures are affected. This study investigates the tolerance of Acartia tonsa nauplii and adults to ammonia, using mortality as the endpoint after 24, 48 and 72 h of exposure. Nauplii were exposed to levels from 0 to 5127 μg NH₃ L^?1 and adults to levels from 0 to 8481 μg NH₃ L^?1. Nauplii NOEC was 30 μg NH₃ L^?1 and LOEC was 81 μg NH₃ L^?1. Adult NOEC was 477 μg NH₃ L^?1 and LOEC was 1789 μg NH₃ L^?1. 50% Lethal Concentrations (LC₅₀) for nauplii of 48 and 72 h was 1257 and 220 μg NH₃ L^?1. LC₅₀ for adults was 2370 (24 h), 972 (48 h) and 770 (72 h). Combining NOEC with excretion rates of NH₄/NH₃ a model was developed to calculate densities in batch cultures. We recommend that for batch cultures of A. tonsa, NH₃ is kept below NOEC for nauplii and that levels of NH₃ together with pH are monitored weekly.     

Chronic exposure of channel catfish, Ictalurus punctatus, to ammonia: Effects on growth and survival

The growth of juvenile channel catfish (Ictalurus punctatus) was reduced in a linear manner during a 31 day growth trial when exposed to concentrations of ammcnia ranging from 48 to 989 μg/l NH₃N (0.31 to 5.71 mg/l NH⁺₄N). On a wet weight basis, growth was reduced by 50% at 517 μg/l NH₃N and the no growth occured at 967 μg/l. The no growth level was 60% of 96-h LC50 value. Mortality was increased significantly at 989 μg/l NH₃N and above. The sublethal effects of ammonia may depend, in part, on the concentrations of NH₄⁺ and and Na⁺ in solution.     

Acute toxicity of ammonia to the early stage-larvae and juveniles of Eriocheir sinensis H. Milne-Edwards, 1853 (Decapoda: Grapsidae) reared in the laboratory

Static-renewal bioassays were performed to evaluate the acute toxicity of ammonia to Eriocheir sinensis (H. Milne-Edwards) at three growing stages, namely zoea-I, zoea-II, and juvenile (0.06 g wet weight per crab). The 24 h LC₅₀ values were 13.3, 20.2, and 109.3 mg (NH₃+ NH₄⁺) 1^?1 (0.47, 0.71, and 3.10 mg NH₃ I^?1), the 48 h LC₅₀ values being 6.8, 10.3, and 60.9 mg (NH₃⁺ NH₄⁺) 1^?1 (0.24, 0.36, and 1.73 mg NH₃1^?1), while the 72 h LC₅₀ values were 5.7, 7.6, and 45.3 mg (NH₃+ NH₄⁺) 1^?1 (0.20, 0.27, and 1.29 mg NH₃ 1^?1) for zoea-I, zoea-II, and juveniles, respectively. The 96 h LC₅₀ value for juveniles was 31.6 mg (NH₃+ NH₄⁺) 1^?1(0.90 mg NH₃1^?1). It was evident that the tolerance to ammonia increased during the same exposure time as the larvae developed to juveniles and decreased with prolonged exposure time. Compared with larvae, juveniles were more sensitive to the fluctuation of ambient ammonia concentrations in the certain range within which partial kills took place. The ‘safe level’ of ammonia based on the 96 h LC₅₀ value and an application factor of 0.1 was 3.16 mg (NH₃+NH₄⁺)1^?1 (0.09 mg NH₃ 1^?1) for juveniles and those for zoea-I and zoea-II were 0.57 and 0.76 mg (NH₃+ NH₄⁺) 1^?1 (0.02 and 0.03 mg NH₃ 1^?1) based on 72 h LC₅₀ values.     

The role of açaí (Euterpe oleracea Mart. 1824) as a chemoprotective agent in the evaluation of antioxidant defence,oxidative damage and histology of juvenile shrimp Litopenaeus vannamei (BOONE, 1931) exposed to ammonia

This study investigated the effects of the use of the inclusion of açaí on the diet of shrimp Litopenaeus vannamei on antioxidant and histopathological responses after exposure to ammonia. The shrimps were fed two experimental diets: control and with 10% of açaí inclusion (W/W), for 35 days. Afterwards, the organisms were exposed at four concentrations of ammonia (0.01‐control; 0.26; 0.48 and 0.91 mg NH₃‐N L^?1) for 96 hr. The total antioxidant capacity (ACAP) of the gills decreased significantly in both diets when exposed to ammonia, whereas in the muscle, the açaí promoted an increase in ACAP. Concomitantly, lipid peroxidation levels increased in the gills and decreased in muscle. After exposure to ammonia, glutathione‐S‐transferase activity increased in hepatopancreas in shrimps fed with açaí facilitating the detoxification of lipid peroxidation by‐products, and the concentration of protein sulfhydryl groups decreased in the gills and muscle of the shrimp of the control diet, evidencing protein damage, an unobserved response in shrimps that received the açaí diet. Histopathological changes decreased in açaí‐fed shrimps about the control diet after exposure to ammonia. It is concluded that açaí mitigated ammonia‐induced histopathological changes, improved the antioxidant defence system (gills and muscle) and attenuated the lipid damage in the muscle.     

Is the use of turmeric in the diet safe for fish?

With this study, we aimed to assess the safety of using Curcuma longa in fish feed. Juvenile Astyanax aff. bimaculatus (0.83 ± 0.04 g) were kept in 24 80‐L aquaria, at a density of 0.5 fish L^?1 for 60 days. Six diets supplemented with 0.0, 20.0, 40.0, 60.0, 80.0 and 100.0 g kg^?1 were evaluated. Histopathological analysis of the gills showed that there was an increasing linear effect of turmeric on the frequency of lamellar aneurysm, hypertrophy of mucous cells, oedema and necrosis. There was a quadratic effect of turmeric supplementation on the frequency of fusion and disruption of secondary lamellae, vacuolation and hypertrophy of the epithelial cells, and the estimated values to minimize these variables were 4.63, 25.93 and 24.46 g kg^?1 respectively. There was a quadratic effect of turmeric on the gills index (I_BRA), and the estimated value to minimize this index was 30.23 g kg^?1. No effect of turmeric on the frequency of histopathological changes and organ index in fish liver was observed. Thus, we conclude that turmeric is not hepatotoxic for A. aff. bimaculatus; however, its use is safe at concentrations up to 30.23 g kg^?1 diet as a function of its gill toxic effect at higher doses.     

Acute responses of juvenile cobia Rachycentron canadum (Linnaeus 1766) to acid stress

Fish are potentially submitted to water acidification when reared in recirculating aquaculture systems. This study evaluated the responses of juvenile cobia Rachycentron canadum after acute exposure to acid water. Juvenile cobia (12.6 ± 0.5 g; 14.2 ± 0.2 cm) were acutely exposed to four pH levels (7.9 (control), 6.5, 6.0, and 5.5). After 24 h of exposure to different pH values, fish were sampled for physiological and histopathological evaluation. Acid water affected physiological parameters and induced morphological histopathologies on gill and skin of juvenile cobia, and these effects were more conspicuous with decreasing pH values. Acid stress induced blood acidosis in juvenile cobia, coupled to a decrease in bicarbonate (HCO₃^?) and saturated O₂ (sO₂) in fish blood. On the other hand, haematocrit, haemoglobin and glucose concentration increased their values (P < 0.01) comparing to control level. Hyperplasia with completely fusion of secondary lamella was observed in all pH treatments (6.5. 6.0 and 5.5), while telangiectasia and proliferation of chloride cells were present for fish exposed to pH 6.0 and 5.5. In skin hyperplasia and hypertrophy of mucous cells, necrosis of these cells for fish exposed to pH 6.0 and 5.5 was observed. The results of this study demonstrate that acute acid water exposition affected physiology and histopathology in juvenile cobia, especially at pH values below 6.5. Accordingly, particular attention must be given to pH during cobia reared in recirculating aquaculture.     

Effect of γ‐aminobutyric acid supplementation on growth performance,endocrine hormone and stress tolerance of juvenile Pacific white shrimp,Litopenaeus vannamei,fed low fishmeal diet

An 8‐week feeding trial was conducted to evaluate the optimum dietary γ‐aminobutyric acid (GABA) level in low fishmeal diet for juvenile Pacific white shrimp, Litopenaeus vannamei. Six practical diets (449 g kg^?1 protein, 87 g kg^?1 lipid) supplemented with graded levels of GABA (0, 50, 100, 150, 200, 250 mg kg^?1) were formulated. Each diet was randomly assigned to triplicate groups of 30 shrimps (approximately 0.44 g), and the shrimps were fed four times a day to apparent satiation. Weight gain and gain rate were significantly increased with the supplementation of GABA (P < 0.05). Hepatosomatic index and survival were also significantly influenced by the dietary GABA levels (P < 0.05) and show a similar trend to those of growth performance. Insulin and neuropeptide Y concentrations in blood were increased with the supplementation of GABA. In total, 150 mg GABA kg^?1‐supplemented diets significantly increased the survival after 12, 24 and 36 h of NH₃ stress, also increased the total antioxidant capacity, total superoxide dismutase, catalase, antihydroxyl radical and phenoloxidase activities before and after the 36 h NH₃ stress compared to basic group. These results clearly indicated that GABA could improve growth performance, antioxidative capacity and resistance against NH₃ stress in L. vannamei, and 150 mg kg^?1 GABA supplementation was suitable for L. vannamei fed with low fishmeal diet.     

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

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