Effect of melatonin on GnIH precursor gene expression in Nile tilapia,Oreochromis niloticus

Sexual maturation and gonadal development of fish is greatly influenced by photic information, an external environmental factor, and melatonin mediates this information to regulate gonadotropin (GTH) secretion and gonadal activation. The relationship between gonadotropin inhibiting hormone (GnIH) and melatonin in fish, however, has not been studied to date. Here, the GnIH expression pattern and daily change of melatonin levels were compared to each other in mature tilapia (body length 16.1 ± 0.2 cm, body weight 77.7 ± 3.43 g), and the effect of melatonin injection on GnIH gene expression was investigated. GnIH gene expression increased at night when the secretion of melatonin increased, whereas gene expression decreased during the day when melatonin secretion decreased. Injecting tilapia intraperitoneally with melatonin increased GnIH gene expression and decreased the expression of gonadotropin releasing hormone (GnRH) and GTH. Furthermore, the injection decreased the 11-KT concentration in male tilapia. These results indicate that melatonin is likely to suppress the hypothalamus-pituitary-gonad (HPG) axis via the action of GnIH in this species.     

Effects of vitamin E on expressions of eight microRNAs in the liver of Nile tilapia (Oreochromis niloticus)

Currently, microRNAs (miRNAs) are known to regulate cellular processes such as apoptosis, differentiation, cell cycle, and immune functions, and their expression can be altered by distinct stress conditions, such as oxidative stress. In immune systems of fish, vitamin E (VE) has a defined role as an antioxidant. In order to understand the molecular mechanism of vitamin E defending from oxidative stress, three groups of juvenile Nile tilapia (Oreochromis niloticus) (initial weight 3.25 ± 0.02 g) were fed to satiation with 3 semi-purified diets containing VE (dl-α-tocopherol acetate) of 0, 50, and 2500 mg/kg supplementation, respectively, with the expressions of eight miRNAs (miR-21, miR-223, miR-146a, miR-125b, miR-181a, miR-16, miR-155 and miR-122) in the liver of tilapia subsequently detected after 8-week growth experiment. Results showed that VE-deficient (0 mg/kg supplementation) decreased the activity of superoxide dismutase (SOD), and decreased the expressions of miR-223, miR-146a, miR-16 and miR-122, while excessive supplementation of VE (2500 mg/kg) decreased SOD activity and increased the expressions of all the eight miRNAs. The targets of the eight miRNAs were further predicated with bioinformatic approach and the possible regulating mechanisms of VE via miRNAs were analyzed. The present study confirmed that the differences in dietary VE affected expression of hepatic miRNAs which may partly demonstrate the molecular mechanism of VE, and the new idea of introducing miRNAs into research will provide the basic data for researches of molecular nutrition.     

Dietary lipid sources affect cold tolerance of Nile tilapia (Oreochromis niloticus)

This study was carried out to evaluate the effects of dietary lipid sources on growth performance, fatty acids composition and cold tolerance of Nile tilapia (Oreochromis niloticus) fingerlings (7.00 ± 0.50 g/fish). The fish were fed four isonitrogenous (28% crude protein), isocaloric (500 kcal/100 g) diets containing four lipid sources; fish oil (FO), corn oil (CO), coconut oil (COCO) or fish oil/ corn oil mixture (1:1 ratio) (oil mix). The diets were offered to the fish at a daily rate of 3% of their body weights (BW), twice a day for two months. After the feeding trial, the fish were exposed to decreasing water temperature from 25 °C until the appearance of death symptoms. The results revealed that FO-based diets (FO and oil mix) produced the best growth rates and feed efficiency, followed by corn oil diet, while COCO resulted in the lowest performance. Fish fed on CO and oil mix showed higher body unsaturated fatty acids (UFA) and lower lethal temperature than those fed on FO- or COCO-based diets. These results indicate that cold shock can modify the lipid metabolism in Nile tilapia by lowering total body saturated fatty acids and raising n-6 and n-3 UFA. This finding suggests that the inclusion of high levels of plant oils in Nile tilapia feeds can enhance their cold tolerance.     

Bone development in the jaw of Nile tilapia Oreochromis niloticus (Pisces: Cichlidae)

East African cichlids have evolved feeding apparatus morphologies adapted to their diverse feeding behaviors. The evolution of the oral jaw morphologies is accomplished by the diversity of bone formation during development. To further understand this evolutionary process, we examined the skeletal elements of the jaw and their temporal and sequential emergence, categorized by developmental stages, using the Nile tilapia Oreochromis niloticus as a model cichlid. We found that chondrogenesis started in Stage 17. The deposition of osteoid for the dermal bones commenced in Stage 18. The uptake of calcium dramatically shifted from the surface of larvae to the gills in Stage 20. The bone mineralization of the skeleton began in Stage 25. These data provide important information regarding the sequential events of craniofacial development in East African cichlids and lay the groundwork for studying the molecular mechanisms underlying adaptation of jaw structure to feeding behavior.     

Deleterious impacts of heat stress on steroidogenesis markers,immunity status and ovarian tissue of Nile tilapia (Oreochromis niloticus)

The water temperature of aquacultures is a primary factor of fish welfare, reproductive patterns, and immunity. To elucidate the molecular and biological processes of the temperature modulation of reproduction and immunity, female Nile tilapia (190 ± 10g) were allocated into five groups following acclimatization (150 females, three replicates, each n = 10). Each group was subjected to various temperatures (28 °C, 30 °C, 32 °C, 34 °C, and 37 °C), the group at 28 °C representing the control. Their serum levels of estradiol, cortisol, and vitellogenin were measured as well as serum triiodothyronine (T3) hormone, thyroxine (T4) hormone, and non-specific immunity (phagocytic and lysozyme activity). In addition, steroidogenic acute regulatory protein (STAR), vitellogenin gene receptor, and heat shock protein 70 (HSP70) gene expression were evaluated. The serum levels of estradiol, cortisol, and vitellogenin markedly declined (P < 0.05) in fish group at higher temperatures. In addition to T3, T4 was significantly affected (P < 0.05) in the control group. The expressions of the STAR gene (steroidogenesis) and vitellogenin receptors were also considerably down-regulated. The histopathological photomicrograph of fish subjected to high water temperature revealed injuries in ovary tissues, demonstrating its harmful effects. The experimental results verified the possible role of water temperature as a main stressor on Nile tilapia’ physiology through modulation of steroidogenesis-related gene expression and immunity.     

Metabolic and molecular responses in Nile tilapia,Oreochromis niloticus during short and prolonged hypoxia

The strictly aquatic breathing Nile tilapia, Oreochromis niloticus is an extremely hypoxia-tolerant fish. To augment our understanding of the effects of hypoxia on anaerobic glycolysis in the Nile tilapia, we studied the effect of short-term for 1 day (trial 1) and long-term for 30 days (trial 2) hypoxia on a selected glycolytic enzymes activity and mRNA expression in liver and white muscle. The hypoxic oxygen concentrations used in the two trials were 2, 1, and 0.5 mg O₂ L^?1 for comparison with a control normoxic group 8 mg O₂ L^?1. The activity of phosphofructokinase (PFK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) in liver and white muscle except liver LDH decreased in trial 1 and increased in trial 2. Assessments of mRNA levels in trial 1 revealed that PFK was downregulated and LDH was upregulated in liver and white muscle, while PK fluctuated between upregulation in liver and downregulation in white muscle. Meanwhile, PK and LDH were upregulated while PFK was similar to control values in both tissues in trial 2. Comet assay results demonstrated an increase in DNA damage that was directly proportional to increasing hypoxic concentrations. This damage was more pronounced in trial 1. This suggests that the Nile tilapia cope better with long-term hypoxic conditions, possibly as an adaptive response.     

Clock-controlled endogenous melatonin rhythms in Nile tilapia (Oreochromis niloticus niloticus) and African catfish (Clarias gariepinus)

The purpose of this work was to investigate the circadian melatonin system in two tropical teleost species characterized by different behavioral habits, Nile tilapia (diurnal) and African catfish (nocturnal). To do so, fish were subjected to either a control photoperiod (12L:12D), continuous light (LL) or darkness (DD), or a 6L:6D photoperiod. Under 12L:12D, plasma melatonin levels were typically low during the photophase and high during the scotophase in both species. Interestingly, in both species, melatonin levels significantly decreased prior to the onset of light, which in catfish reached similar basal levels to those during the day, demonstrating that melatonin production can anticipate photic changes probably through circadian clocks. Further evidence for the existence of such pacemaker activity was obtained when fish were exposed to DD, as a strong circadian melatonin rhythm was maintained. Such an endogenous rhythm was sustained for at least 18 days in Nile tilapia. A similar rhythm was shown in catfish, although DD was only tested for four days. Under LL, the results confirmed the inhibitory effect of light on melatonin synthesis already reported in other species. Finally, when acclimatized to a short photo-cycle (6L:6D), no endogenous melatonin rhythm was observed in tilapia under DD, with melatonin levels remaining high. This could suggest that the circadian clocks cannot entrain to such a short photocycle. Additional research is clearly needed to further characterize the circadian axis in teleost species, identify and localize the circadian clocks, and better understand the environmental entrainment of fish physiology.     

Circadian Rhythms of Locomotor Activity in the Nile Tilapia Oreochromis niloticus

Behavioral rhythms of the Nile tilapia were investigated to better characterize its circadian system. To do so, the locomotor activity patterns of both male and female tilapia reared under a 12:12 h light-dark (LD) cycle were studied, as well as in males the existence of endogenous rhythmicity under free-running conditions (DD and 45 min LD pulses). When exposed to an LD cycle, the daily pattern of activity differed between individuals: some fish were diurnal, some nocturnal, and a few displayed an arrhythmic pattern. This variability would be typical of the plastic circadian system of fish. Moreover, reproductive events clearly affected the behavioral rhythms of female tilapia, a mouth-brooder teleost species. Under DD, 50% (6 of 12) of male fish showed circadian rhythms with an average period (τ) of 24.1±0.2 h, whereas under the 45 min LD pulses, 58% (7 of 12) of the fish exhibited free-running activity rhythms with an average τ of 23.9±0.5 h. However, interestingly in this case, activity was always confined to the dark phase. Furthermore, when the LD cycle was reversed, a third of the fish showed gradual resynchronization to the new phase, taking 7–10 days to be completely re-entrained. Taken together, these results suggest the existence of an endogenous circadian oscillator that controls the expression of locomotor activity rhythms in the Nile tilapia, although its anatomical localization remains unknown.     

The impacts of invasive Nile tilapia (Oreochromis niloticus) on the fisheries in the main rivers of Guangdong Province,China

Nile tilapia (Oreochromis niloticus) is one of the most widespread invasive fish species, and this species has successfully established populations in the major rivers of Guangdong Province, China. Field surveys and manipulative experiments were conducted to assess the impacts of Nile tilapia on fisheries. We determined that the increase of Nile tilapia in these rivers not only affects the CPUE (catch-per-unit-per-effort) of the fish community and native fish species but also reduces the income of fishermen. In the manipulative experiments, we observed that the growth of native mud carp decreased in the presence of Nile tilapia. Our results suggest that the invasion of Nile tilapia negatively affected the fishery economy and native fish species, and suitable control measurements should be taken.     

Development of the embryo, larva and early juvenile of Nile tilapia Oreochromis niloticus (Pisces: Cichlidae). Developmental staging system

We described the developmental stages for the embryonic, larval and early juvenile periods of Nile tilapia Oreochromis niloticus to elucidate sequential events of craniofacial development. Craniofacial development of cichlids, especially differentiation and morphogenesis of the pharyngeal skeleton, progresses until about 30 days postfertilization (dpf). Because there is no comprehensive report describing the sequential processes of craniofacial development up to 30 dpf, we newly defined 32 stages using a numbered staging system. For embryonic development, we defined 18 stages (stages 1-18), which were grouped into seven periods named the zygote, cleavage, blastula, gastrula, segmentation, pharyngula and hatching periods. For larval development, we defined seven stages (stages 19-25), which were grouped into two periods, early larval and late larval. For juvenile development until 30 dpf, we defined seven stages (stages 26-32) in the early juvenile period. This developmental staging system for Nile tilapia O. niloticus will benefit researchers investigating skeletogenesis throughout tilapia ontogeny and will also facilitate comparative evolutionary developmental biology studies of haplochromine cichlids, which comprise the species flocks of Lakes Malawi and Victoria.     

Duplication of neuropeptide Y and peptide YY in Nile tilapia Oreochromis niloticus and their roles in food intake regulation

In vertebrates, the neuropeptide Y (NPY) family peptides have been recognized as key players in food intake regulation. NPY centrally promotes feeding, while peptide YY (PYY) and pancreatic polypeptide (PP) mediate satiety. The teleost tetraploidization is well-known to generate duplicates of both NPY and PYY; however, the functional diversification between the duplicate genes, especially in the regulation of food intake, remains unknown. In this study, we identified the two duplicates of NPY and PYY in Nile tilapia (Oreochromis niloticus). Both NPYa and NPYb were primarily expressed in the central nervous system (CNS), but the mRNA levels of NPYb were markedly lower than those of NPYa. Hypothalamic mRNA expression of NPYa, but not NPYb, decreased after feeding and increased after 7-days of fasting. However, both NPYa and NPYb caused a significant increase in food intake after an intracranial injection of 50 ng/g body weight dose. PYYb, one of the duplicates of PYY, had an extremely high expression in the foregut and midgut, whereas another form of duplicate PYYa showed only moderate expression in the CNS. Both hypothalamic PYYa and foregut PYYb mRNA expression increased after feeding and decreased after 7-days of fasting. Furthermore, the intracranial injection of PYYb decreased food intake, but PYYa had no significant effect. Our results suggested that although the mature peptides of NPYa and NPYb can both stimulate food intake, NPYa is the main endogenous functional NPY for feeding regulation. A functional division has been identified in the duplicates of PYY, which deems PYYb as a gut-derived anorexigenic peptide and PYYa as a CNS-specific PYY in Nile tilapia.     

Genetic characterization of four strains of Nile tilapia (Oreochromis niloticus L.) using microsatellite markers

Four domesticated strains of Nile tilapia (Oreochromis niloticus L.) were genetically characterized using 14 microsatellite markers and 64 animals per strain. Two strains, Chitralada (AIT) and International Development Research Centers (IDRC) were obtained from the AIT institute, Bangkok, Thailand. The GIFT strain (5th generation) came from NAGRI, Thailand, and the GÖTT strain was supplied by the University of Göttingen, Germany. The average numbers of alleles per marker were 5.0 (GÖTT), 5.4 (AIT), 5.6 (IDRC) and 7.5 (GIFT). Private alleles were found at all markers with the exception of two. No fixation of alleles was found at any marker. Population differentiation, F_ST, was 0.178 (great genetic differentiation) and confirmed grouping of the animals in strains. The expected level of heterozygosity ranged from 0.624 to 0.711, but the observed level of heterozygosity significantly deviated from the expected level in three strains. This was probably because of small population size. Moderate to great genetic differentiation was found between strains. A phylogenetic tree reflected the strains known histories. Application of the Weitzman approach showed that all strains have added value for the total genetic diversity and thus should be retained.     

Molecular cloning and gene expression of Foxl2 in the Nile tilapia, Oreochromis niloticus

A Foxl2 cDNA was cloned from the Nile tilapia ovary by RT-PCR and subsequent RACE. Alignment of known Foxl2 sequences from vertebrates confirmed the conservation of the Foxl2 open reading frame and protein sequences, especially the forkhead domain and C-terminal region, while some homopolymeric runs of amino acids are found only in mammals but not in non-mammalian vertebrates. RT-PCR revealed that Foxl2 is expressed in the tilapia brain (B), pituitary (P), gill, and gonads (G), with the highest level of expression in the ovary, reflecting the involvement of Foxl2 in B-P-G axis. Northern blotting and in situ hybridization also revealed an evident sexual dimorphic expression pattern in the gonads. Foxl2 mRNA was mainly detected in the granulosa cells surrounding the oocytes. The ovarian expression of Foxl2 in tilapia begins early during the differentiation of the gonads and persists until adulthood, implying the involvement of Foxl2 in fish gonad differentiation and the maintenance of ovarian function.     

Effect of water temperature,feeding frequency,and protein percent in the diet on water quality,growth and behavior of Nile tilapia Oreochromis niloticus (Linnaeus, 1758)

This study evaluated how water temperature (26, 28, and 30°C), number of meals per day (one or two meals), and protein percent in diet (20, 25 and 30%) impact growth performance, biometric indices, and feeding behavior of Nile tilapia, Oreochromis niloticus. Fish were randomly allocated into 18 equal replicate groups. Higher final body weight was observed in fish reared at 30°C and fed one meal per day containing 30% crude protein. Better weight gain, weight gain %, feed conversion ratio, specific growth rate, and condition factor were recorded in fish reared at 26°C and fed one meal per day containing 30% protein. The best length weight relationship was obtained in fish reared at 26°C and fed one meal per day containing 30% crude protein. Shorter feeding duration and duration of appetite inhibition latency were recorded in fish reared at 30°C, fed one meal per day, and given a diet containing 30% protein. The highest proactivity was recorded in fish reared at 30°C, received one meal per day, and with 25% crude protein in their diet. Conclusively, rearing Nile tilapia at 26–30°C with a lower feeding frequency (one meal/day) and a 30% crude protein diet achieved better performance and feeding behavior.     

Effect of some common West African farm-made feeds on the oxygen consumption and ammonia excretion rates of Nile tilapia,Oreochromis niloticus

The effects of two common West African farm-made feeds on postprandial metabolism were assessed in adult Nile tilapia (Oreochromis niloticus) by quantifying the oxygen consumption rates (MO₂) and total ammonia-nitrogen (TAN) excretion over a 24-h period. Measurements followed the ingestion of a single meal of a fishmeal-based control diet and two diets containing 30% copra (CM) or palm kernel meal (PKM) inclusions by groups of 15 adult O. niloticus per tank. The mean net MO₂ (postprandial oxygen consumption corrected for routine metabolic rate) during digestion for the different tilapia groups varied narrowly between 112.2 ± 9.9 and 129.9 ± 20.4 mg O₂ kg^?1 h^?1 with the fish fed the CM diet recording a significantly lower (p = 0.04) net MO₂ response relative to the other two diets. Net TAN excretion rates of the different dietary groups varied between 3.4 ± 1.4 and 4.4 ± 1.6 mg TAN kg^?1 h^?1. Under the standardized experimental conditions, copra and PKMs appeared to be promising candidates as partial replacements to fishmeal in tilapia diets as far as rates of oxygen consumption and ammonia-nitrogen excretion are concerned.     

Efficacy of Nile tilapia (Oreochromis niloticus) juveniles and spinosyns bioinsecticides against aquatic stages of Culex pipiens: An experimental study

The mosquito Culex pipiens is the most widely distributed dipteran species in all regions of Egypt and the principal vector of Wuchereria bancrofti and certain arboviruses in human beings. For controlling C. pipiens vector, biological tools (e.g., larvivorous fish and bioinsecticides) are more potent and safer options to the environment, human beings, and beneficial organisms than chemical pesticides. The efficiency of O. niloticus juveniles as predatory fish species and two bioinsecticides, spinosad 24% and spinetoram 12%, was investigated against the C. pipiens developmental stages in the laboratory. The first trial evaluated the predatory efficacy of small-sized O. niloticus (2.1–2.6 cm; 250–315 mg) and large-sized O. niloticus (2.5–3.2 cm; 250–315 mg) against the 3rd larvae and pupae of C. pipiens. This is the first report in Egypt confirming the predation potential of O. niloticus as efficient predatory fish against the immature C. pipiens. Large-sized O. niloticus predated a greater number of 3rd of C. pipiens larvae and pupae than the small-sized ones. Furthermore, the daily consumption of C. pipiens larvae by small- and large-sized O. niloticus was significantly higher than the pupae. The second trial assessed the toxicity efficacy of spinosad 24% and spinetoram 12% against C. pipiens larvae and pupae. The results confirmed that the tested bioinsecticides showed higher potency toward C. pipiens larvae than pupae after exposure for 24 h and 48 h. Spinosad was more toxic toward 3rd C. pipiens larvae (LC₅₀ = 0.013 and 0.003 mg/L) and pupae (LC₅₀ = 320.69 and 44.28 mg/L) than spinetoram after 24 and 48 h. Herein, O. niloticus juveniles (as promising native predatory fish) and spinosyns bioinsecticides were more effective against C. pipiens in the larval stage than in the pupal stage. In conclusion, Nile tilapia juveniles and biorational compounds, spinosad 24% and spinetoram 12%, might be considered as promising and favorable environmental biological agents for controlling C. pipiens in Egypt. However, further trials are needed to investigate the potential of these agents in the control of this mosquito vector under field conditions.     

尼罗罗非鱼Orexin前体基因的克隆、组织分布及其在摄食调控中的表达

该文采用RT-PCR和cDNA末端快速扩增技术(rapid-amplification of cDNA ends,RACE)的方法,从尼罗罗非鱼(Oreochromis niloticus)下丘脑总RNA中获得了尼罗罗非鱼Orexin前体基因的cDNA全长序列。该cDNA全长648bp,其中开放阅读框的长423bp,编码Orexin前体蛋白为140个氨基酸,包括37个氨基酸的信号肽、43个氨基酸的Orexin-A、28个氨基酸的Orexin-B和末尾32个氨基酸组成的功能不详的多肽。采用Real-time PCR技术对尼罗罗非鱼Orexin前体基因的组织表达模式以及在摄食前后、饥饿和再投喂状态下的表达量变化进行了研究。结果显示,在脑部和外周等18个组织中都检测到了Orexin前体基因的表达,其中在下丘脑中表达量最高;在摄食前后,尼罗罗非鱼Orexin前体基因的表达量显著低于在摄食状态中;饥饿2、4、6和8d后,Orexin前体基因在下丘脑中的表达量与正常投喂组相比均显著升高,饥饿4d再投喂后,表达量又恢复至正常水平。这些结果表明,Orexin在尼罗罗非鱼摄食中可能有着重要的调节作用。     

Effect of bisphenol A toxicity on growth performance,biochemical variables,and oxidative stress biomarkers of Nile tilapia,Oreochromis niloticus (L.)

Bisphenol A (BPA) is one of the industrial chemical compound which is used in the production of polycarbonate plastics and epoxy resins. BPA is used throughout the world and it could enter the aquatic ecosystems causing serious problems. To evaluate the potential effects of BPA toxicity on Nile tilapia, Oreochromis niloticus (L.) performance, its lethal concentration (LC₅₀) was determined and it was 13.13 µg/L. After that, fish (33.9 ± 0.55 g/fish) were exposed to 0.0, 1.64, or 3.28 µg/L of BPA for 6 weeks after which growth performance, biochemical variables, and oxidative defense system were assessed. The results showed that fish growth and feed intake were significantly reduced as BPA levels increased with no significant difference in fish survival. Total protein, albumin, globulin, and acetylcholine esterase decreased significantly; meanwhile, aspartate transferase, alanine transferase, alkaline phosphatase, uric acid, and creatinine increased significantly with exposure to BPA in a dose dependent manner. Furthermore, malondialdehyde value and the activities of superoxide dismutase and catalase increased significantly; while glutathione peroxidase and glutathione S‐transferase decreased significantly as BPA levels increased. In conclusion, BPA exposure in aquatic environment deteriorated fish performance and health causing liver and kidney dysfunction. Thus, fish exerted oxidative defense enzymes as a protection tool against BPA toxicity.