Measurements of auditory sensitivity in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> by the auditory brainstem response technique and cardiac conditioning method

ABSTRACT:   Recently, the non-invasive auditory brainstem response (ABR) technique has been applied to the determination of hearing sensitivity in fish. The technique has some advantages over, and methodological differences from, the classical techniques. The auditory thresholds of common carp were measured by the ABR technique. The classical method was applied to measure cardiac response by electrocardiogram (ECG) after conditioning with electric shock. The most sensitive frequency obtained by ABR was 505 Hz, and by ECG, 1000 Hz. The shapes of the audiogram obtained by ABR and ECG were similar, though threshold levels in both audiograms differed from each other. Furthermore, audiograms obtained by using behavioral techniques were compared with those taken by the ABR and ECG techniques to assess the validity of the ABR technique for testing fish hearing sensitivity. The measured threshold levels increased in the order of behavioral, ABR, and ECG techniques, in the range below 1000 Hz.     

Method for underwater measurement of the auditory brainstem response of fish

ABSTRACT:   The auditory brainstem response (ABR) of fishes is commonly measured by bringing the heads of the fishes out of the water in a small tank. However, this method is inapplicable to experiments for large fishes that are economically important in large spaces such as the sea or in a large tank. This paper describes a method of recording, the ABR for fishes in water, without exposing the fish heads to air, by using a waterproof, insulated electrode. To evaluate the effectiveness of this method, the goldfish Carassius auratus was investigated, and the ABR waveform and auditory thresholds measured in water were compared with those measured on the surface. Both ABR waveforms and auditory thresholds showed similar trends between the two methods. The underwater ABR method is useful to measure the auditory thresholds of larger fish in natural or on-site environments such as the sea, net enclosures and large aquaria in which precise positioning of the fish is not possible. However, more improvement is needed to apply this method to large fishes.     

Audiogram measurement based on the auditory brainstem response for juvenile Japanese sand lance <Emphasis Type="Italic">Ammodytes personatus</Emphasis>

In this study, the auditory thresholds for juvenile Japanese sand lance Ammodytes personatus were measured based on its auditory brainstem response (ABR). The amplitude of the ABR waveforms to a sound stimulus were larger than that of the electric background noise caused by general brainwaves and myogenic signals after the averaging procedure. Japanese sand lance responded to low frequency sounds between 128 Hz and 512 Hz with a sound pressure level of 115–125 dB. As the test frequency decreased, so did the auditory threshold level, and the level was about 116 dB at 128 Hz and 181 Hz. These results indicate that Japanese sand lance can detect low frequency sound but are less sensitive than other fish species. These high thresholds are probably caused by the lack of a swim bladder.     

Hemato-immunological responses of Heros severus fed diets supplemented with different levels of Dunaliella salina

In this study, the effects of oral administration of different levels of Dunaliella salina (a natural β-carotene source) on growth parameters, immunological and hematological indices, as well as skin carotenoids, of Heros severus were investigated. One hundred and eighty H. severus weighing 27 ± 0.5 g were divided randomly into four groups in triplicate (15 fish in each replicate). Groups 1–4 received food supplemented with 0, 50, 100 and 200 mg kg^?1 D. salina powder, respectively. After 6 weeks, the growth parameters were compared among the groups. Blood samples were taken from each group, and hematological parameters including red blood cell count (RBC), white blood cell count (WBC), hematocrit (PCV), hemoglobin (Hb) and immunological indices (serum and mucus lysozyme and bactericidal activity, resistance against Aeromonas hydrophila infection) as well as carotenoid content of skin were evaluated. Results showed that some growth indices increased significantly in fish fed with 100 and 200 mg kg^?1 D. salina-supplemented food (P < 0.05). Although serum lysozyme activity was increased in fish fed with food supplemented with 100 and 200 mg kg^?1 D. salina (P < 0.05), no significant change was observed in serum and mucus bactericidal activity and mucus lysozyme activity among the groups (P > 0.05). Most of the hematological parameters such as WBC, RBC, PCV and Hb significantly increased in D. salina-treated fish compared with controls (P < 0.05). Mortality induced after challenge with A. hydrophila in 200 mg kg^?1 D. salina-treated fish was 36.67 %, which significantly decreased compared with control (P < 0.05). Skin carotenoid content in all D. salina treatments was statistically higher than that of control (P < 0.05). Conclusively, D. salina as a food additive can affect positively the growth, immunological and hematological parameters of H. severus.     

胭脂鱼听觉阈值研究

利用听性脑干反应技术, 研究了胭脂鱼(Myxocyprinus asiaticus)的听觉阈值。通过插入皮下的电极记录了10尾胭脂鱼的短纯音听觉诱发电位。结果显示, 在系统最大声强下, 胭脂鱼的听频范围为100~5 000 Hz, 其中对100~2 000 Hz的声音敏感度较高, 最敏感的频率为800 Hz, 听觉阈值约为69.8 dB。随着声强的减小, 胭脂鱼的听性脑干反应波形最大, 振幅减小, 潜伏期延长。将10尾胭脂鱼的阈值平均, 得到了胭脂鱼的听性脑干反应听力图。胭脂鱼的听力图呈“U”形, 属于典型的动物听力曲线。研究胭脂鱼的听觉阈值对胭脂鱼物种保护具有重要意义, 同时可为评估噪声对胭脂鱼的影响提供数据支持。

     

Spotlined sardine Sardinops melanostictus listensto 1-kHz sound by using its gas bladder

ABSTRACT:   The auditory characteristics of pelagic fish have received lessattention compared to those of benthic or freshwater fish species,even though underwater sounds are considered to affect their behavior.Audiograms of five spotlined sardines were obtained using the auditorybrainstem response technique in which the evoked potential on theskin covering the head of the fish was measured. The auditory brainstemresponse is a non-invasive electrophysiological method that enablesthe auditory threshold level of fragile species, such as spotlinedsardines, to be measured. The gas bladder of the spotlined sardinehas been considered as possibly contributing to an enhancement intheir hearing. In the test, the sound absorption profile of sardineswas also measured. The frequency at the lowest auditory thresholdlevel (i.e. the most sensitive frequency) and absorption were foundat 1024 Hz and 1040 Hz, respectively. The spotlinedsardine was found to be sensitive to sounds at relatively higherfrequency ranges compared to other seawater fishes and the gas bladderis seen to play a significant role in detecting sound.     

Physiological response,blood chemistry profile and mucus secretion of red sea bream (<Emphasis Type="Italic">Pagrus major</Emphasis>) fed diets supplemented with <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> under low salinity stress

Environmental stressors caused by inadequate aquaculture management strategies suppress the immune response of fish and make them more susceptible to diseases. Therefore, efforts have been made to relieve stress in fish by using various functional feed additives in the diet, including probiotics. The present work evaluates the effects of Lactobacillus rhamnosus (LR) on physiological stress response, blood chemistry and mucus secretion of red sea bream (Pagrus major) under low salinity stress. Fish were fed four diets supplemented with LR at [0 (LR0), 1 × 10² (LR1), 1 × 10⁴ (LR2) and 1 × 10⁶ (LR3) cells g^?1] for 56 days. Before stress, blood cortisol, urea nitrogen (BUN) and total bilirubin (T-BIL) showed no significant difference (P > 0.05), whereas plasma glucose and triglyceride (TG) of fish-fed LR2 and LR3 diets were significantly lower (P < 0.05) than those of the other groups. Plasma total cholesterol (T-CHO) of fish-fed LR3 diet was significantly (P < 0.05) lower than that of the other groups. Furthermore, total plasma protein, mucus myeloperoxidase activity and the amount of mucus secretion were significantly enhanced in LR-supplemented groups when compared with the control group (P < 0.05). After the application of the low salinity stress test, plasma cortisol, glucose, T-CHO and TG contents in all groups showed an increased trend significantly (P < 0.01) compared to the fish before the stress challenge. However, plasma total protein and the amount of secreted mucus showed a decreased trend in all groups. On the other hand, BUN, T-BIL and mucus myeloperoxidase activity showed no significant difference after exposure to the low salinity stress (P > 0.05). In addition, the fish that received LR-supplemented diets showed significantly higher tolerance against low salinity stress than the fish-fed LR-free diet (P < 0.05). The physiological status and the detected immune responses, including total plasma protein and mucus myeloperoxidase activity in red sea bream, will provide a more comprehensive outlook of the effects of probiotics to relieve stress in fish.     

Immunomodulatory effect of cimetidine in common carp (Cyprinus carpio L.)

The main indication of cimetidine is being H₂-receptor antagonist, but studies suggest that cimetidine may also act as a non-specific stimulant of cell-mediated immunity and immunomodulator. In order to determine the immunomodulatory effect of dietary intake of cimetidine in the common carp (100 ± 10 g), subjects were fed diets containing 0 (control), 50, 100 and 200 mg cimetidine kg^?1 of dry diet for a period of 6 weeks. TLC and NBT assays were significantly (P < 0.05) stimulated in cimetidine-supplemented groups displaying the highest value in 200 mg kg^?1 group. A decrease (P < 0.05) in cortisol and ACH50 value was recorded in fish treated with cimetidine. Serum protein, albumin and serum globulin levels were not significantly changed. The findings of the present investigation suggest that the incorporation of cimetidine in the diet of common carp enhances the non-specific immunity.     

Effect of replacement of fish oil with camelina (Camelina sativa) oil on growth, lipid class and fatty acid composition of farmed juvenile Atlantic cod (Gadus morhua)

Camelina (Camelina sativa) oil was tested as a replacement for fish oil in diets for farmed Atlantic cod (Gadus morhua). Camelina differs from other plant oilseeds previously used in aquaculture with high lipid (40 %), α-linolenic acid (40 %), antioxidants and low proportions of saturated fats. Dietary treatments were fed to cod (19 g fish^?1 initial weight) for 9 weeks and included a fish oil control (FO), 40 % (CO40) and 80 % (CO80) replacement of fish oil with camelina oil. There was no effect of replacing fish oil with camelina oil included at levels up to 80 % on the growth performance. Cod fed CO80 stored more lipid in the liver (p < 0.01), including more neutral lipid (p < 0.05) and triacylglycerol (p < 0.05). Cod fed CO80 decreased in total polyunsaturated fatty acids (PUFAs) in muscle compared to CO40 and FO (p < 0.05), increased in monounsaturated fatty acids (p < 0.01), decreased in total ω3 fatty acids (FO > CO40 > CO80; p < 0.01) and increased in total ω6 fatty acids (FO < CO40 < CO80; p < 0.01). In the liver, long-chain (LC) PUFA such as 20:4ω6, 20:5ω3, 22:5ω3 and 22:6ω3 decreased when fish oil was removed from the diet (p < 0.05), and increased in 18-carbon fatty acids (p < 0.01). Camelina oil can reduce the amount of fish oil needed to meet lipid requirements, although replacing 80 % of fish oil reduced LC PUFAs in both tissues. A comparison of BF₃ and H₂SO₄ as catalysts to transmethylate cod liver and muscle lipids revealed small but significant differences in some fatty acid proportions.     

心电图技术和听性脑干反应技术在测定鱼类听觉性能上的比较

简要介绍了ECG技术、ABR技术、鱼类听觉特性及ECG技术、ABR技术在测定鱼类听觉性能上的比较。对于两种测定鱼类听觉性能的方法,分别从原理、测定方法、所用仪器、适用类型(鱼类)、优缺点方面进行比较,重点介绍了同一实验用两种方法测量所得结果分析。     

The potential of soya oil industry products as oil alternatives in rainbow trout (Oncorhynchus mykiss) diet

This study investigated the effects of different soya oil products on growth, nutrient digestibility, and fatty acid composition of muscle in rainbow trout. The products’ ingredients were soap stock, fat powder, crude oil, and refined soya oil as a control. Four experimental diets were formulated by addition of 12 % of each of the four ingredients to a basal diet. Replacement of refined soya oil by the soya oil products did not influence growth performance until 8 weeks (P > 0.05). This replacement, however, reduced fat digestibility in rainbow trout (83.7 vs. 73.3–79.8 %; P < 0.05). Dietary inclusions of fat powder and crude oil led to larger values of protein digestibility than those of refined soya oil and soap stock (P < 0.05). The compositions of saturated and monounsaturated fatty acids of the muscle were similar, but linoleic acid concentration (LA, 18: 2n-6) increased in fish muscle fed refined soya oil compared to the other diets (37 vs. 30–31 %; P < 0.05). Arachidonic acid (ArA, 20: 4n-6) concentration was higher (P < 0.05) in fish fed both fat powder and soap stock than those fed the other diets. In conclusion, refined soya oil can be replaced by soya oil products in rainbow trout diet with no negative impacts on the growth. However, fish fed on diet containing refined soya oil had higher concentration of LA and lower density of ArA. It appears that rainbow trout is capable of digesting soya oil products though the digestibility rate reduced slightly with an increase in saturated fatty acid content.     

Modulation of antioxidant defense and immune response in zebra fish (<Emphasis Type="Italic">Danio rerio</Emphasis>) using dietary sodium propionate

The present study explores the effect of dietary sodium propionate on mucosal immune response and expression of antioxidant enzyme genes in zebra fish (Danio rerio). Six hundred healthy zebra fish (0.42 ± 0.06 g) supplied, randomly stocked in 12 aquariums and fed on basal diets supplemented with different levels of sodium propionate [0 (control), 5, 10 and 20 g kg^?1] for 8 weeks. At the end of the feeding trial, mucosal immune parameters (TNF-α, IL-1β, Lyz), antioxidant enzyme (SOD, CAT) as well as heat shock protein 70 (HSP70) gene expression were measured. The results revealed feeding on sodium propionate significantly up-regulated inflammatory response genes (TNF-α, IL-1β, Lyz) in a dose-dependent manner (P < 0.05). However, antioxidant enzyme genes significantly down-regulated in the treated group compared with control (P < 0.05). Also, HSP70 gene expression was higher in the liver of fish fed the basal diet and deceased with elevation of sodium propionate levels in the diet. These results showed beneficial effects of dietary sodium propionate on mucosal immune response as well as the antioxidant defense of zebra fish.     

Effects of exogenous non-starch polysaccharide-degrading enzymes in diets containing <Emphasis Type="Italic">Gracilaria lemaneiformis</Emphasis> on white-spotted snapper <Emphasis Type="Italic">Lutjanus stellatus</Emphasis> Akazaki

A feeding experiment was conducted to investigate the effects of exogenous non-starch polysaccharide (NSP)-degrading enzymes in diets containing Gracilaria lemaneiformis (GL) on growth performance and digestive enzyme activities of white-spotted snapper Lutjanus stellatus Akazaki (initial mass 8.0 ± 0.1 g). A basal diet (D0) containing a mixed protein source (fish meal, soybean meal and GL meal) was used as the control. Two diets supplemented with 0.5 g (D1) and 1 g (D2) exogenous NSP-degrading enzymes per kilogram of diet were formulated. Each diet was assigned to triplicate groups of fish in a total of nine floating sea cages (270 fish, 30 fish per cage). After a 60-day feeding trail, significantly higher weight gain, specific growth ratio and feed efficiency ratio were observed in fish fed D2 diet compared to those of control (P < 0.05). Body lipid, moisture and ash contents were not significantly affected by NSP-degrading enzyme supplementation, but significantly higher protein content was noticed in fish fed D2 diet compared to that of control (P < 0.05). The amylase activity in the stomach and intestine was significantly higher in fish fed D2 diet (P < 0.05), but no significant differences were observed in pepsin and lipase activities in the stomach or trypsin and lipase activities in the intestine between all treatments. The results suggested that addition of 1 g kg^?1 NSP-degrading enzymes in diet could efficiently improve seaweed feed utilization and growth performance of white-spotted snapper fish.     

Effect of prebiotic konjac mannanoligosaccharide on growth performances,intestinal microflora,and digestive enzyme activities in yellow catfish,Pelteobagrus fulvidraco

In the present study, konjac mannanoligosaccharide (KMOS) was evaluated as a prebiotic in yellow catfish. The fish were fed with diets containing KMOS in four concentrations: 0 g kg^?1 (C), 1.0 g kg^?1 (KM1), 2.0 g kg^?1 (KM2), and 3.0 g kg^?1 (KM3) for 49 days, respectively. Another group fed with diets containing 3.0 g kg^?1 yeast cell wall mannanoligosaccharide (MOS) (M3) was set as positive control. The results indicated that fish receiving the diets supplemented with KMOS or MOS showed higher relative gain rate (RGR), specific growth rate (SGR), and lower feed conversion ratio (FCR) with significantly differences (P < 0.05) than those fed with the basal diets. Moreover, fish receiving the diets with 2.0 g kg^?1 KMOS inclusion showed higher RGR, SGR, and lower FCR (P < 0.05) than that feeding the diets supplemented with 3.0 g kg^?1 MOS. The quantities of Bifidobacterium spp. were significantly increased (P < 0.05). Meanwhile, Escherichia coli and Aeromonas spp. were significantly reduced (P < 0.05) in the fish-feeding diets with 2.0 g kg^?1 KMOS supplement. Compared with the control group, the significantly enhancement of protease and amylase activity (P < 0.05) in intestine and pancreas was observed in fish fed with diets containing KMOS or MOS. Collectively, an optimum level of KMOS inclusion in diets could modulate intestinal microflora, induce digestive enzyme activity, and improve the growth performance of yellow catfish significantly.     

Sound generated by a payao and comparison with auditory sensitivity of jack mackerel <Emphasis Type="Italic">Trachurus japonicus</Emphasis>

Sound generated by a payao, an anchored bamboo fish aggregating device, is believed to be attractive to fish; but until now, there is no available record of payao-generated sound. This study presents payao-generated sound recorded by a hydrophone at water depths of 5, 10 and 15 m from a fixed distance of 3 m relative to the payao, and compares the sound with the auditory sensitivity of jack mackerel Trachurus japonicus measured at discrete frequencies from 100 to 2000 Hz using the auditory brainstem response protocol. A consistent peak appeared in the sound spectrum at 49 Hz and showed an increasing sound pressure level with depth, which suggests that payao sound may come from the anchor rope. However, the contribution of the bamboo raft can not yet be discounted. The hearing threshold curve indicated that the most sensitive frequency range in jack mackerel is from 92.1 dB at 800 Hz to 111.0 dB at 200 Hz. These results show that the dominant frequency range of payao sound does not correspond with the high sensitivity frequency range of fish hearing.     

Daily rhythms of blood glucose differ in diurnal and nocturnal European sea bass (Dicentrarchus labrax L.) undergoing seasonal phase inversions

Sea bass change their feeding rhythms from diurnal to nocturnal in winter, returning to diurnal feeding in spring. Despite behavioral data, the physiological changes that take place during such changes remain unexplored. In this paper, blood glucose rhythms of European sea bass with diurnal/nocturnal self-feeding rhythms were investigated during phase inversions of their feeding behavior (in winter and spring) when both diurnal and nocturnal fish coexist. Blood glucose showed daily variations in both seasons (ANOVA, p < 0.03), fitting a cosine function (COSINOR, p < 0.05) in all cases, except in diurnal fish in spring. The average blood glucose levels of nocturnal fish in winter (2.67 ± 0.09 mmol/l, mean ± SEM) were significantly (t test, p < 0.01) higher than in spring (2.20 ± 0.08 mmol/l), while they were similar (~2.25 mmol/l) in diurnal fish in both seasons. These findings revealed for the first time insights into the seasonal physiological changes that accompany changes in behavioral rhythms in diurnal and nocturnal sea bass.     

Effect of feeding rotifers enriched with taurine on the growth and survival of larval amberjack Seriola dumerili

The effect of feeding rotifers enriched with taurine on the growth performance and survival of larval amberjack Seriola dumerili was investigated. Rotifers were enriched with a commercial taurine supplement at four levels (0, 200, 400, and 800 mg/l). The larvae were fed the enriched rotifers in triplicate from 3 days post-hatch for 7 days under static conditions. The average taurine contents of the rotifers were 1.5, 2.7, 4.2, and 7.2 mg/g dry matter, respectively. The growth of the fish fed rotifers enriched with the taurine supplement at 800 mg/l was significantly (P < 0.05) improved compared with that of the fish fed the rotifers without taurine enrichment. The survival rate improved proportionally up to a taurine supplement level of 400 mg/l, but no significant differences in survival were observed among treatments. The fraction of the larvae with inflated swim bladders did not vary significantly between treatments. Taurine content in the whole fish body increased with the taurine level in the rotifers. These results suggest that taurine enrichment of rotifers is an effective method of enhancing the growth of amberjack larvae.     

Effects of stocking density of Labeo rohita on survival, growth and production in cages

Labeo rohita (139.92 ± 0.76 mm/24.33 ± 0.45 g) was reared for 92 days in floating square cages (10 m² area, 1.5 m height) in a pond (2 ha) at six stocking densities (5, 7.5, 10, 15, 20 and 25 fish m^?2) each with 3 replicates. Fish were fed daily once in the morning with rice polish and groundnut oil cake (1:1) in dough form at 3 % of the total body weight. Survival ranged from 96 to 100 % in different stocking densities. Final average body weight, average body weight gain, mean daily body weight gain and SGR decreased (P < 0.05) with increasing stocking density. Conversely, final biomass, biomass gain and FCR increased (P < 0.05) with increasing stocking density. The highest growth rate of fish could be achieved up to 60 days at 5 fish m^?2 and 92 days at other densities. The reduced growth rate at 10–25 fish m^?2 for 60 days of culture indicated that stress is related to size and density of the fish, suggesting that utmost care is required to reduce the stress at high densities. Maximum production and profit was observed at the highest stocking density. Non-lethal levels of water and soil qualities at different sites (cage premises, and 20 and 200 m away from cage area) suggested that cage aquaculture could be done safely covering 0.9 % of pond area. Production of advanced fingerlings in cages was found a viable alternative to their culture in pond.     

Long-term effects of intraperitoneal injection of estradiol-17β on the growth and physiology of juvenile stellate sturgeon Acipenser stellatus

Juvenile stellate sturgeon Acipenser stellatus were intraperitoneally injected with estradiol-17β (E₂; 0 and 5 mg/kg fish) to investigate the possibility of sex reversal and also determine the changes in biochemical parameters. Five-month-old fish (40.9 ± 1.1 g) were injected every 3-week interval during a 190-day trial. At the termination of the experiment, final weight and other growth parameters including weight gain and specific growth rate, hepatosomatic and viscerosomatic indices were not affected by repetitive injection of E₂. Hematological features of E₂-treated fish showed significant reductions in number of red blood cells, hemoglobin concentration, hematocrit value and mean corpuscular hemoglobin (P < 0.05), but no significant changes were observed in number of white blood cells, mean corpuscular volume and mean corpuscular hemoglobin concentration (P > 0.05). Calcium, phosphorus, glucose, triacylglycerol, cholesterol, total protein and estradiol concentrations were significantly increased in fish injected with E₂ (P < 0.001). Plasma progesterone and testosterone levels were noticeably lower in fish injected with 5 mg/kg E₂ rather than the control fish (P < 0.001). Histological observations of gonads showed that all fish injected with 5 mg/kg E₂ apparently feminized, while 66.6 % of the control group was female. These results revealed that the injection of E₂ is an effective method for feminization of stellate sturgeon without having significant inhibitory effects on growth and survival.     

Effect of treatment with probiotics as water additives on tilapia (Oreochromis niloticus) growth performance and immune response

A feeding trial was conducted for 40 days to delineate the effect of treatment with probiotics as water additives on tilapia (Oreochromis niloticus) growth performance and immune response. About 360 juveniles were randomly distributed into four treatment groups, each with three replicates. Different probiotics (T-1, Bacillus subtilis B10; T-2, Bacillus coagulans B16; T-3, Rhodopseudomonas palustris G06) were added to the water of tanks at final concentration of 1 × 10⁷ cfu ml^?1 every 2 days, with no probiotic added to control tanks. At the end of the feeding trial, fish treated with B. coagulans B16 (T-2) and R. palustris G06 (T-3) had significantly (P < 0.05) higher final weight, daily weight gain, and specific growth rate compared with those treated with B. subtilis B10 (T-1) and those without probiotics (control). The highest (P < 0.05) content of total serum protein was found in T-2 compared with that in T-1, T-3, and the control. However, albumin concentration and albumin/globulin ratio were not affected by the probiotics treatments. Compared with the control, probiotic supplementation remarkably improved activities of superoxide dismutase and catalase (P < 0.05). T-2 fish exhibited higher average myeloperoxidase activity than the control, T-1, and T-3 groups. Regarding serum lysozyme content in tilapia, assays showed no difference (P > 0.05) among the treatment groups. Furthermore, probiotics treatments remarkably increased respiratory burst activity compared with control, with T-2 showing higher values than T-1 and T-3. This indicated that treatment with probiotics, B. coagulans B16 and R. palustris G06, as water additives could be used to enhance immune and health status, thereby improving growth performance of O. niloticus.