Physiological responses and HSP70 mRNA expression in GIFT tilapia juveniles,Oreochromis niloticus under short‐term crowding

Physiological responses and HSP70 mRNA expression to short‐term crowding were tested in juveniles genetically improved farmed tilapia (Oreochromis niloticus) (70.43 ± 4.43 g). Fish were kept at control group (5 g L^?1) and stress groups (low density 10 g L^?1, medium density 40 g L^?1 and high density 70 and 100 g L^?1) for 48 h. Each density was tested in triplicates. The effects of a short‐term exposure on the physiological responses of fish were determined before stress (0 h) and at 6, 12, 24 or 48 h post‐crowding. There was a significant increase in serum cortisol, total protein, lysozyme and aspartate aminotransferase (AST) activities in all stressed groups at 24 h post‐crowding (P < 0.05). Serum glucose and alanine aminotransferase (ALT) of 100 g L^?1 stressed group were lower than that of the control after 48 h of crowding stress (P < 0.05). The levels of serum triglyceride and cholesterol in crowding stress with high density were significantly decreased compared with the control group at 48 h. The mRNA expression data showed that hepatic HSP70 mRNA levels were markedly elevated at all stressed groups. HSP70 mRNA levels of 70 and 100 g L^?1 stressed groups decreased at 48 h compared with the 24 h post‐crowding. The protective ability of HSP70 was limited.     

Stable isotope analysis to quantify contributions of supplementary feed in Nile Tilapia Oreochromis niloticus (GIFT strain) aquaculture

Nile tilapia (Oreochromis niloticus) is currently one of the most farmed freshwater fish and contributes significantly to total global aquaculture production. The genetically improved strain of O. niloticus (GIFT) was introduced to Papua New Guinea (PNG) in 1999 to improve food and income security. The high cost and low availability of commercial fish feed hinder the growth of GIFT farming in PNG. Stable carbon and nitrogen isotopes were used to determine the role of supplementary and natural food sources in the diet of GIFT in pond‐based aquaculture. Two treatments were used: treatment 1 was daily feeding, and treatment 2 was weekly feeding, each with three replicates. Isotopic analysis of muscle tissue and all potential food sources showed that pellet feed contributed 7% to the growth of GIFT in daily‐fed ponds and 33% in the weekly‐fed ponds. Highly enriched δ¹⁵N values for chicken manure, compared to depleted values for GIFT and other natural food sources in both treatments, clearly indicate insignificant contributions of this input to production. After 90 days of cultivation, the average final body weight of GIFT receiving daily feed inputs was 134 g (average 19 cm), while for weekly‐fed it was 92 g (17 cm). The feed conversion ratio (FCR) was poor (6.4:1) in the daily‐fed GIFT ponds compared to a better, and preferable, FCR (1:1) in the weekly‐fed ponds. The findings of this study show that pelleted feed was not the major contributor to the growth of GIFT. Genetically improved farmed tilapia aquaculture should focus on enhancing natural food availability for fish production.     

Combined effects of temperature and salinity on yolk utilization in Nile tilapia (Oreochromis niloticus)

The combined effects of temperature and salinity on the yolk utilization of sac fry in Nile tilapia (Oreochromis niloticus) were investigated using central composite experimental design and response surface approach. Based on the preliminary trials, temperature was determined to range from 22 to 34°C, and salinity ranging from 2 ppt to 10 ppt. The utilization was mensurated in terms of yolk sac volume. Results showed that the linear effects of temperature and salinity on the yolk utilization was significant (P < 0.01); the quadratic effects of and the interaction between the two factors were significant (P < 0.05); temperature was more important than salinity in influencing the yolk utilization. The model equation of yolk sac volume towards temperature and salinity was established. From those high R² values, the model had excellent goodness of fit to experimental data and could be applied for predictive purpose. What with the production cost, it is suggested that the temperature/salinity combination, i.e. 28–30°C/4–6 ppt, be employed during the period of sac fry rearing, in which the yolk utilization was on average 98.6%.     

Moderate salinities enhance growth performance of Nile tilapia (Oreochromis niloticus) fingerlings in the biofloc system

The biofloc technology (BFT) has recently gained attention as an economic and environmentally sustainable system for aquaculture. The use of BFT with salinized water could be used to minimize the negative effects of nitrogenous waste and improve the growth performance of Nile tilapia. This work evaluated the growth performance, survival, gill lesions, and fillet composition of tilapia fingerlings (Oreochromis niloticus) reared during 70 days with different salinities (0, 4, 8, 12, and 16 g/L) in a biofloc previously developed. The results indicated that the use of mature bioflocs alone was sufficient to avoid fingerling mortality due to nitrite peaks. Moreover, the salinized water, especially between 4 and 8 g/L (maximum points around 6 g/L), can be recommended in BFT to improve the growth performance of tilapia in the initial culture phase. The salinity level that was evaluated did not affect fillet composition nor the occurrence of gill lesions. The total ammonia nitrogen concentration was lower in freshwater than in salinized water (p < 0.05). Nitrite peaks in salinities of 12 and 16 g/L appeared later and were higher in comparison to other treatments. Considering minimal water exchange in BFT, the use of salinized water in this system may be environmentally viable.     

Utilization of Caridina nilotica (Roux) meal as a protein ingredient in feeds for Nile tilapia (Oreochromis niloticus)

The effects of replacing fish meal with Caridina nilotica as a protein ingredient on growth performance, nutrient utilization, carcass, proximate composition and economic benefits in Nile tilapia (Oreochromis niloticus) culture was evaluated. Replacement of the FM with C. nilotica was done at 25%, 50%, 75% and 100% (D25, D50, D75 and D100) and the substitution effects was compared with the control diet (D0, 0% C. nilotica). After 140 days of culture, the best growth performance, nutrient utilization and economic benefits occurred in fish groups fed diets with 25% C. nilotica inclusion. However, growth performance in fish fed diets D50 and D75 were comparable with the control (P > 0.05). At 100% substitution level of FM with C. nilotica, the growth performance and fish survival was lower than control. Protein and lipid contents in the fish and their digestibilities were highest in diet D25 and decreased with increasing levels of substitution of FM with C. nilotica. This study demonstrate that utilization of local protein sources (C. nilotica) can be effectively used to replace up to 75% of FM in the diets without compromising growth performance, survival, nutrient utilization and economic benefits in O. niloticus culture.     

Dietary lycopene supplementation on Nile Tilapia (Oreochromis niloticus) juveniles submitted to confinement: effects on cortisol level and antioxidant response

This study evaluates the inclusion of lycopene in the commercial diet for Nile Tilapia exposured to a stressor. For 60 days, fish diet was supplemented with lycopene (600 mg kg^?1 diet) and at the end, subjected to confinement for 90 min. Zootechnical parameters (weight gain, specific growth rate and intake), hormonal (cortisol level), biochemical {enzyme activity and lipid peroxidation level [thiobarbituric acid (TBARS)]} and physical (instrumental colour) parameters were also evaluated. Supplementation with lycopene did not alter zootechnical parameters. In unconfined fish, catalase (CAT) activity was reduced in the white muscle of fish fed with lycopene; however, in red muscle, it was not altered. In fish under confinement, plasma cortisol increased as well as CAT, glutathione reductase (GR) and lactate dehydrogenase (LDH) activity in both muscles. Lipid peroxidation of the fillet also displayed higher values. However, the lycopene supplementation abolished the effects of confinement available through cortisol levels, remaining unchanged in the above parameters, activity of CAT, GR and LDH in both muscles and TBARS content of the fillet; in addition, it did not induce alteration of fillet coloration. Inclusion of lycopene in commercial feed reduces the impact of confinement and does not alter the zootechnical parameters; additionally, no modification of the fillet colour from juvenile Nile tilapia was observed.     

Dualistic feeding pattern of Nile tilapia (Oreochromis niloticus,L.) reared under different self‐feeding system conditions

Nile tilapia (Oreochromis niloticus) is one of the most important cultured species worldwide. The research aims to clarify the feeding characteristics, such as daily feeding pattern of Nile tilapia using a self‐feeding technique. The feeding pattern was conducted under two rearing conditions, indoor treatments under a controlled light regime (LD 12:12) and constant water temperature (25°C), and outdoor treatments under natural conditions which consisted of duplicate trials with two periods each. The outdoor treatment was carried out from early summer through late autumn in Mie, Central Japan. Daily self‐feeding activity of Nile tilapia in indoor treatments was nearly daytime feeding pattern, synchronizing with the given photoperiod (24 hr). However, the self‐feeding activity in the outdoor experiments from early summer to early autumn was almost daytime feeding pattern, but it became less clear and shifted to a nighttime feeding profile in late autumn. The results revealed that Nile tilapia has a dualistic capacity for demand‐feeding both in light and dark phases. These results might have been caused by the seasonal change in light intensity and/or water temperature. Information obtained from the self‐feeding experiments enables us to identify the influence of environmental changes on the physiological condition of farmed fish through their expression of appetite.     

Effects of dissolved oxygen on the growth performance,haematological parameters,antioxidant responses and apoptosis of juvenile GIFT (Oreochromis niloticus)

This study investigated the effects of different dissolved oxygen (DO) levels on the growth performance, antioxidant response and apoptosis of juvenile GIFT (genetically improved farmed tilapia, Oreochromis niloticus). GIFT were fed with five DO levels (1, 2, 3, 4 and 5 mg/L) for 60 days, and the results showed that the final body weight, weight gain rate, specific growth rate and crude protein and crude lipid contents of the fish muscle increased at 5 mg/L DO. The activities of the antioxidant and digestive enzymes were significantly up‐regulated with increasing DO levels. However, the haemoglobin content, number of red blood cells, malondialdehyde content, transaminase activities, glucose content and lactic acid levels decreased at higher DO levels. Furthermore, the cardiomyocyte apoptotic index was significantly decreased with increasing DO levels. Our results show that 5 mg/L DO improved growth performance, promoted antioxidant enzyme activities and reduced liver damage in GIFT.     

Dietary supplementation of Tribulus terrestris extract improves growth and reproductive performances of the male Nile tilapia (Oreochromis niloticus)

Effects of dietary supplementation of Tribulus terrestris (TT) and 17α‐methyl testosterone (MT) on growth performance and reproductive efficiency of male Nile tilapia, Oreochromis niloticus (N = 75, initial weight 60 ± 3.251 g), were evaluated. Five experimental treatments in triplicate were fed on basal diet (30% crude protein) (control group, CG), basal diet containing 60 mg MT (MT group) and basal diet containing 250, 500 and 750 mg/kg diet TT (TT250, TT500 and TT750 groups), respectively, for consecutive 45 days. Results revealed that the fish received TT in diet showed better growth performance (final body weight, weight gain, feed conversion ratio, specific growth rate, testes weight and gonadosomatic index) compared with MT and CG (p < .05), as well as for haemoglobin content and red blood cell count (p < .05). The highest level of testosterone, 11‐keto testosterone, sperm concentration and vitality was recorded in TT750 followed by TT500, TT250 and MT groups, while the lowest value was recorded in the CG (p < .05). The highest seminiferous tubule diameter was recorded in TT750 followed by TT500, TT250 and CG, while the lowest diameter was recorded in the MT group (p < .05). Fish received TT showed normal dilated seminiferous tubule filled with large amount of free sperms within their lumen in a dose related effect. In conclusion, male Nile tilapia dietary supplementation with different inclusions of Tribulus terrestris extract specially 500 and 750 mg/kg diet improves growth performance, health condition, semen quality and reproductive efficiency without any harmful effect on water quality.     

Relationship of RNA/DNA ratio to somatic growth of Nile tilapia juveniles (Oreochromis niloticus) under joint effects of temperature and salinity

Tilapia juveniles are a very important life stage, and reliably assessing their growth performance is of prime importance in aquaculture production. The suitability of the RNA/DNA as a bioindicator for growth evaluation in tilapia has not yet been reported. In this study, we examined suitability of RNA/DNA ratio for assessing growth of tilapia juveniles and variation in the ratio under the concurrent influences of temperature and salinity using the central composite design and response surface methodology. Results showed that under our experimental conditions, the synergistic effects of temperature and salinity on the RNA/DNA ratio were highly significant (P < 0.01), and the ratio varied with the two factors in a nonlinear fashion; the temperature × salinity interaction was detected at 1% significance level, and had a negative effect on the RNA/DNA ratio particularly at higher temperatures. Temperature was found to have a greater influence on RNA/DNA ratios than salinity. A model of the relation between RNA/DNA, temperature and salinity was developed, with an R² greater than 98%. The linear relationships of absolute growth, cumulative growth and specific growth of tilapia juveniles to RNA/DNA ratio all showed an R² of greater than 95%. Our results indicate that the RNA/DNA ratio was a responsive and accurate bioindicator for evaluating the growth performance of juvenile tilapia. These ratio‐based models are neither temperature‐ nor salinity‐dependent, and could be readily applicable to freshwater or brackish water aquaculture within temperature–salinity combinations ranging from 16 to 36°C and 0–16 ppt.     

Effects of ferulic acid on growth performance,immunity and antioxidant status in genetically improved farmed tilapia (Oreochromis niloticus) fed oxidized fish oil

A study was conducted to characterize the effects of dietary oxidized fish oil on the growth performance, immunity and antioxidant status of genetically improved farmed tilapia (Oreochromis niloticus) and to determine the role of ferulic acid on the oxidative damage induced by the oxidized fish oil. The tilapia (13.73 ± 0.31 g) were fed four experimental diets containing untreated (peroxide value, POV: 2.2 meq/kg) and highly oxidized (POV: 120.6 meq/kg) fish oil either with or without ferulic acid (0 or 400 mg/kg) supplementation for 12 weeks. From the results, the oxidized fish oil treatments increased antioxidant enzyme activities and MDA values but decreased the weight gain and the immunological parameters in tilapia. Meanwhile, the serum biochemical indices were significantly affected by the oxidized fish oil. Besides, the addition of ferulic acid partially counteracted the free radical‐induced damage and improved the health status of tilapia. In conclusion, the oxidized fish oil may induce oxidative stress, destroy liver, dysregulate lipid metabolism as well as reduce non‐specific immunity, and eventually result in growth inhibition of tilapia. The ferulic acid supplementation partially offset the negative effects of the oxidized fish oil on tilapia.     

氨氮与拥挤胁迫对吉富品系尼罗罗非鱼幼鱼生长和肝脏抗氧化指标的联合影响

采用中心复合试验设计(CCD)和响应曲面方法(response surface methodology,RSM),探讨了氨氮(0.02 ～ 2.00 mg/L)和养殖密度(1～5尾/10 L)对吉富罗非鱼幼鱼生长和肝脏抗氧化指标的联合影响.结果表明,本试验条件下,氨氮和养殖密度的一次与二次效应对特定生长率有显著影响(P＜0.05),随着氨氮或养殖密度的上升,特定生长率呈先上升后下降的变化.氨氮与养殖密度之间存在互作效应(P＜0.05),氨氮浓度为0.02 ～0.20 mg/L,养殖密度在1～2尾/10 L时,幼鱼特定生长率较高;而氨氮浓度高于0.20 mg/L,养殖密度在3尾/10 L左右时,生长速度较快.肝脏丙二醛(MDA)含量随氨氮浓度和养殖密度的上升而上升,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活力呈先上升后下降的变化.氨氮与养殖密度的一次效应对MDA含量和两种酶活力均有显著影响(P＜0.05),二次效应对两种酶活力的表达有极显著影响(P＜0.01);氨氮与养殖密度对CAT活力有互作效应,高浓度氨氮与高养殖密度环境会抑制SOD和CAT活力的表达.因子与响应值间二次多项回归方程的决定系数分别达到0.972 4、0.913 2、0.938 9和0.969 2(P＜0.01),可用于预测;氨氮效应对生长和抗氧化酶活力的影响较养殖密度明显.建议在罗非鱼的养殖过程中合理安排好养殖密度,保持溶氧充足,降低氨氮胁迫,提高罗非鱼的生长与抗病力.     

Development of attenuated erythromycin‐resistant Streptococcus agalactiae vaccine for tilapia (Oreochromis niloticus) culture

Streptococcus agalactiae is an important pathogen in fish, causing great losses of intensive tilapia farming. To develop a potential live attenuated vaccine, a re‐attenuated S. agalactiae (named TFJ‐ery) was developed from a natural low‐virulence S. agalactiae strain TFJ0901 through selection of resistance to erythromycin. The biological characteristics, virulence, stability and the immunization protective efficacy to tilapia of TFJ‐ery were determined. The results indicated that TFJ‐ery grew at a slower rate than TFJ0901. The capsule thickness of TFJ‐ery was significantly less (p < 0.05) than TFJ0901. When Nile tilapia were intraperitoneally (IP) injected with TFJ‐ery, the mortality of fish was decreased than that injected with TFJ0901. The RPS of fish immunized with TFJ‐ery at a dose of 5.0 × 10⁷ CFU was 95.00%, 93.02% and 100.00% at 4, 8 and 16 weeks post‐vaccination, respectively. ELISA results showed that the vaccinated fish produced significantly higher (p < 0.05) antibody titres compared to those of control at 2 or 4 weeks post‐vaccination. Taken together, our results suggest that erythromycin could be used to attenuate S. agalactiae, and TFJ‐ery is a potent attenuated vaccine candidate to protect tilapia against S. agalactiae infections.     

Determination of histological and genotoxic effects of formalin on Nile tilapia (Oreochromis niloticus L.)

Formalin (37–40% formaldehyde) is one of the most effective, widely used chemical in pisciculture for its antiparasitic, antifungal and prophylactic activities. It is used in paints, cleaning products and textile industry, as well. Genotoxic and histological effects of sublethal formalin exposure on Nile tilapia (Oreochromis niloticus) are investigated in this study. The semi‐static acute test was employed. One‐tenth, one‐fifth and half of the 96‐h LC₅₀ value, 148 mg L^?1, were used as sublethal exposure concentrations: 15, 30 and 75 mg L^?1 for 24, 48, 96 and 168 h respectively. The micronucleus test was applied to investigate the genotoxic effects on fish erythrocytes at the end of predetermined exposure periods and all tissues of the fish were sampled for histological examination. The micronucleus frequencies increased significantly in all exposure groups when compared with their control groups in a dose and time dependent manner (P < 0.05). Results of histological examination showed no histopathological findings in the gonads, heart, skeletal muscle, spleen and intestine tissues after sublethal formalin exposure. Passive hyperaemia of liver tissue in varying proportions, hydropic degeneration, fatty degeneration in high concentrations, branchitis on gill tissues in varying proportions, epithelial lifting, telangiectasia, bleeding in kidney and hyperaemia were found in Nile tilapia exposed to formalin at different concentrations and durations. As a result, it is thought that formalin leads to tissue damage and shows genotoxic effects even at 15 mg L^?1 concentration in O. niloticus.     

CD40 induces an antimicrobial response against the intracellular pathogen Streptococcus agalactiae in Nile tilapia,Oreochromis niloticus

Streptococcus agalactiae is a Gram‐positive facultative intracellular bacterium that leads to severe economic loss of tilapia worldwide. Previous studies demonstrated that CD40 contributes to host protection against intracellular injection. In this study, CD40 was characterized from Nile tilapia (Oreochromis niloticus), named OnCD40. Sequence analysis showed that open reading frame of OnCD40 was 933 bp, containing a single peptide, a transmembrane domain and four cysteine‐rich domains. The qRT‐PCR revealed that OnCD40 was expressed in all examined tissues with the most abundant ones in spleen and thymus. After S. agalactiae stimulation, the expression of OnCD40 was significantly induced in most of the detected organs. Moreover, OnCD40‐overexpressing fish elicited significant protection against subsequent S. agalactiae challenge; approximately 10000‐fold fewer bacteria were detected in spleen of OnCD40‐overexpressing fish in comparison with control fish. Thus, CD40 had protecting function in Nile tilapia against intracellular pathogens.     

Impact of sublethal di‐n‐butyl phthalate on the aquaculture fish species Nile tilapia (Oreochromis niloticus): histopathology and oxidative stress assessment

Phthalates such as di‐n‐butyl phthalate (DBP) and their esters are widely used plasticizers, their ubiquitous presence in daily life, inevitably leads to their restricted use due to important environmental pollution and health impacts and endocrine disruption potential. The aim of this study was to examine the effects of a sublethal concentration of 10 mg L^?1 DBP on haematocrit (HCT) values, gills and liver histology, malondialdehyde (MDA, 2‐thiobarbituric acid‐TBA reactivity) and reduced glutathione (GSH) levels in gills and liver tissue as oxidative stress biomarkers in the aquaculture fish species Nile tilapia (Oreochromis niloticus) after 24 (DBP‐24) and 96 (DBP‐96) h exposure. No differences were found between per cent HCT values in the 24 h exposure groups (P > 0.05). Response of antioxidant defence systems in liver and gill tissues of the fish were dependent on exposure duration and changed to a higher extent during 96 h. MDA levels in liver tissue increased in DBP treated fish in comparison to the control fish. However, the differences between the exposure and control groups were not significant (P > 0.05). A statistically significant decrease (P > 0.05) was recorded in gill MDA levels in the DBP‐96 group when compared to the control and DBP‐24 groups. The liver GSH levels were unchanged in the DBP treated fish. However, GSH levels were significantly lower in the gill tissue of the DBP‐96 group. Exposure to DBP caused several degenerative changes in the histology of gill and liver tissue. Gills displayed hyperaemia, epithelial lifting, oedema, talengiectasia, epithelial hyperplasia and fusion of secondary lamellae, whereas in liver several circulatory anomalies (hyperaemia, blood congestion and sinosoid dilatation) and vacuolization of hepatocytes were observed. Histopathological results demonstrated that the gills were more affected than the liver perhaps due to their direct contact with DBP.     

低温胁迫对尼罗罗非鱼水通道蛋白(Aquaporin 1，AQP1)基因(AQP1)表达的影响

以经过连续多代抗寒选育获得的尼罗罗非鱼(Oreochromis niloticus)耐寒品系为实验材料, 克隆了尼罗罗非鱼水通道蛋白基因(AQP1)的cDNA序列, 并采用实时荧光定量PCR分析方法探讨了低温胁迫对罗非鱼AQP1基因表达水平的影响。序列分析结果显示, 尼罗罗非鱼AQP1 cDNA长度为1 098 bp, 包含36 bp的5′端非翻译区序列、783 bp开放阅读框(ORF)和279 bp的3′非翻译区序列, 编码261个氨基酸。氨基酸序列比对表明, 各物种间AQP1序列的同源性较高(96%~59%)。聚类分析表明, 尼罗罗非鱼首先与同属鲈形目丽鱼科的斑马拟丽鱼(Maylandia zebra)聚在一起, 再与其他硬骨鱼类聚为一类。实时荧光定量PCR 结果表明: 在水温从30℃逐渐降到10℃过程中, 肌肉和肝组织AQP1基因的表达量从20℃开始均逐渐下调。其中肌肉组织中, 与30℃时的表达量相比, 耐寒品系在20℃时表达量大幅度下调, 当水温为15℃时, 其表达量下调幅度为22.16倍。当水温降到10℃时, 其表达量下调了107.73倍。而对照组AQP1基因的表达量在15℃时, 下调幅度为5.38倍。当水温达到10℃时, 其表达量下调幅度为11.30倍。结果分析显示, 耐寒品系与对照组AQP1基因在低温条件下的表达量存在显著性差异(P<0.05)。而在肝组织中, 当水温降到15℃时, 耐寒品系和对照组的表达量下调幅度分别为3.38倍、1.42倍。水温降到10℃时, 其表达量上调幅度分别为18.85倍、9.01倍。结果分析表明, 低温条件下耐寒品系与对照组AQP1基因在肝组织中的表达差异不显著(P>0.05)。结果表明, AQP1表达对温度敏感, 耐寒品系较高的耐寒能力与其在低温条件下的肌肉组织大幅上调表达有关。由此可见, AQP1基因在罗非鱼低温适应过程中发挥重要作用, 是潜在的研究罗非鱼耐寒机制的候选基因之一, 本研究为进一步研究罗非鱼的耐寒分子机制和开展鱼类抗逆育种提供参考。

     

Evaluation of hydrolysed rubber seed meal as a dietary protein source for Nile tilapia Oreochromis niloticus L.

This study evaluated the use of hydrolysed rubber seed meal as a locally grown alternative feed ingredient to reduce the use of fishmeal within formulated feeds for tilapia. Five experimental diets were prepared by formulating different inclusion levels of hydrolysed rubber seed meal (0%, 25%, 50%, 75% and 100% of plant‐based protein contribution). Dried and defatted rubber seed meal was hydrolysed by adding rumen digesta liquid to the ingredient. For digestibility measurement, chromic oxide was added to each diet as an inert marker. The digestibility trial was conducted over a 14‐day period by collecting faecal matter from the rearing tanks. In addition, a growth trial was conducted over a period of 50 days using four replicates per dietary treatment. In both experiments, all fish were fed three times daily to apparent satiation. At the end of growth trial, the inclusion level of hydrolysed rubber seed meal up to 50% of plant‐based protein contribution did not give any significant difference compared to the control diet in terms of feed efficiency, specific growth rate and protein retention parameters. In the digestibility study, fish fed diets containing hydrolysed rubber seed meal up to a 50% of plant‐based protein inclusion level did not give any significant difference compared to the control diet in terms of protein and dry matter apparent digestibility. Based on the result of this growth and digestibility study, up to a 50% of plant‐based protein contribution of hydrolysed rubber seed meal can be used within formulated tilapia feeds without any adverse effect.     

Effects of putative dietary probiotics from the gut of milkfish (Chanos chanos) on the growth performance and intestinal enzymatic activities of juvenile Nile tilapia (Oreochromis niloticus)

Enzymes synthesized by the gut microbiome are recognized as vital for assisting host metabolism. This study was conducted to screen and identify bacterial isolates with enzyme-producing abilities from the intestine of milkfish Chanos chanos. Overall, 114 isolates were collected, of which 58% showed clear zones indicating various enzyme activities via plate-based assays. Specifically, 65 isolates were amylolytic, 1 isolate was cellulolytic, and 18 isolates exhibited both activities. The two most promising isolates—CC8 and CC27—were further subjected to spectrophotometric enzyme assays to determine their amylase, cellulase and protease activities. Results showed that amylase, cellulase and protease activities (6.00 ± 0.01, 0.52 ± 0.02 and 18.62 ± 0.16 U mg^?1 protein, respectively) were higher in CC8 than in CC27. Using 16S rRNA gene sequencing, the isolates were putatively identified as Vibrio sp. and Bacillus cereus respectively. At a suspension (mL) to feed (g) ratio of 3:25, dietary supplementation of putative probiotics was carried out to evaluate the growth performance, survival and enzymatic activities of juvenile Nile tilapia Oreochromis niloticus. Results revealed that probiotic-supplemented fish exhibited better growth performance—final average body weight, weight gain, specific growth rate and feed conversion ratio—along with increased intestinal enzymatic activities compared with the control (p < 0.05). Survival rates ranged from 80% to 90% and were statistically similar in all treatments (p > 0.05). This study concludes that the application of Vibrio sp. CC8 and Bacillus cereus CC27, supplemented either in monostrain or multistrain preparation, can promote growth and enzymatic activities in juvenile Nile tilapia.