Establishing the optimal essential amino acid ratios in juveniles of Nile tilapia (Oreochromis niloticus) by the deletion method

One of the strategies used to improve fish production is lowering the feed costs and the environmental impact by reducing dietary protein content. Using the deletion method, we determined the optimal amino acid (AA) ratio for Nile tilapia (Oreochromis niloticus) (body weight 20 g). Eleven experimental diets and four replications, taken two at a time, distributed in a randomized block design were used. For this trial, a balanced diet (BD) was formulated. Ten other diets were formulated by the deletion method, in which the BD diet was adjusted to result in a reduction of 45% of the test amino acid. Fish were fed three times a day for 57 days. Groups of fish at the beginning and at the end of the experiment were euthanized for further determination of the carcass nitrogen (N) composition. The optimal ratio of each AA was derived by dividing the requirement for each AA by the requirements for lysine. The essential AA ratios, expressed relative to lysine (=100), were methionine 64, threonine 93, tryptophan 24, arginine 125, histidine 34, isoleucine 57, leucine 96, valine 76 and phenylalanine 101. Our findings might be used to design strategies aimed at reducing the production costs of Oreochromis niloticus.     

Sex reversal in Nile tilapia (Oreochromis niloticus Linnaeus) by androgen immersion

Two experiments were conducted to investigate the efficacy of three androgens applied through immersion treatments on the sex ratio of Nile tilapia (Oreochromis niloticus L.) fry. In experiment 1, 14 days post‐hatching (DPH) larvae were exposed to a single immersion treatment in 17α‐methyltestosterone (MT), 17α‐methyldihydrotestosterone (MDHT) or 17α‐ethynyltestosterone (ET) at 200, 600 and 1800 μg L^?1 over 4 h (130 larvae per treatment). In experiment 2, Nile tilapia larvae were exposed to the higher androgen concentration (1800 μg L^?1) applied as either a single immersion (14 DPH) or double immersion (10 and 14 DPH) over 4 h (125 larvae per treatment). Change in sex proportion within each experiment as well as between experiments was analysed by the chi‐square test. In experiment 1, MT, MDHT and ET were equally effective in significantly increasing the proportion of males when applied at 1800 μg L^?1 (86.0%, 90.0% and 86.7% respectively). At 200 μg L^?1 none of the androgens altered sex ratio. At 600 μg L^?1, only MDHT slightly, but significantly skewed the sex ratio towards males (73.0%). In experiment 2, a single immersion treatment at 14 DPH (1800 μg L^?1) significantly increased the proportion of males, but at this time the response was significantly hormone dependent (MDHT, 100.0%; MT, 91.6%; ET, 76.9%). When compared with a single immersion, two‐immersion treatments significantly increased the proportion of males in the MT‐treated group (from 91.6% to 98.3%), decreased the proportion of males in the MDHT group (from 100.0% to 93.4%) and had no significant effect the ET‐treated group (change from 76.9% to 82.5%). The overall comparison of the sex ratio among same treatments from different experiments (a single immersion in 1800 μg L^?1) was not significantly different.     

Molecular and histopathological characterization of Photobacterium damselae in naturally and experimentally infected Nile tilapia (Oreochromis niloticus)

Mass mortality has occurred among cultured Nile tilapia, Oreochromis niloticus, on fish farms in Manzala, Dakahlia province, Egypt, in the summer season, 2019. Moribund fish were reported with deep ulcers, septicaemic lesions and sampled for bacterial isolation. In this study, most isolates were subjected to bacteriological examination, antibiotic sensitivity test, 16S rRNA gene sequencing and histopathological examination. Following isolate identification, intraperitoneal challenge of Nile tilapia with a bacterial suspension 2 × 10⁶ CFU/ml was performed. Samples from liver, spleen and kidney were collected for histological and biochemical analysis. The results showed a high similarity (99%) to Photobacterium damselae strains using phylogenetic analysis of 16S rRNA. P. damselae exhibited resistance to amoxicillin and erythromycin, as well it was highly sensitive to chloramphenicol and doxycycline. Moreover, haemorrhage, oedema, hemosiderosis and melanomacrophage activation in the liver and head kidney of infected fish were detected by light and electron microscopy. Also, significant higher levels of CAT and SOD in the spleen and head kidney, as well as the serum levels of NO were observed in experimentally challenged O. niloticus, compared to the control fish. Our data identified P. damselae for the first time from infected Nile tilapia, describing its sensitivity to a variety of antibiotics, histopathological alterations and oxidative stress impact, and it could be useful indicators for understanding P. damselae pathogenesis, which might provide a preventive efficacy for P. damselae.     

Glycerol inclusion in the diet of Nile tilapia (Oreochromis niloticus) juveniles

The present study evaluated the effects of inclusion of glycerol in the diet of Nile tilapia juveniles on growth performance, biochemical changes in blood, and carcass composition. We used 300 Nile tilapia juveniles with an average initial weight of 29.15 ± 8.40 g and 11.55 ± 0.87 cm in length, distributed in 20 fiberglass tanks with a capacity of 250 L. The experiments followed a completely randomized design with five treatments and four replications during 79 days. The animals were fed diets containing four concentrations of glycerol (25 g kg^?1, 50 g kg^?1, 75 g kg^?1 and 100 g kg^?1) and a control diet without glycerol. HDL was the only biochemical parameter, that showed statistically different (P < 0.05) results; it was higher in the groups fed with 0 and 75 g kg^?1 glycerol compared to the other groups. No significant difference was observed in the results from the carcass composition of tilapia juvenile fed with the different glycerol levels, except for lipids (P < 0.05), which showed the highest values in fish fed with 50 g kg^?1 glycerol and the lowest in fish fed with 100 g kg^?1. Glycerol can be used in fish diets as an energy supplement without causing damage to growth performance or to the biochemical and carcass composition of Nile Tilapia juveniles.     

Detection of tilapia lake virus (TiLV) infection by PCR in farmed and wild Nile tilapia (Oreochromis niloticus) from Lake Victoria

Tilapia lake virus disease (TiLVD) has emerged to be an important viral disease of farmed Nile tilapia (Oreochromis niloticus) having the potential to impede expansion of aquaculture production. There is a need for rapid diagnostic tools to identify infected fish to limit the spread in individual farms. We report the first detection of TiLV infection by PCR in farmed and wild Nile tilapia from Lake Victoria. There was no difference in prevalence between farmed and wild fish samples (p = .65), and of the 442 samples examined from 191 fish, 28 were positive for TiLV by PCR. In terms of tissue distribution, the head kidney (7.69%, N = 65) and spleen (10.99%, N = 191), samples had the highest prevalence (p < .0028) followed by heart samples (3.45%, N = 29). Conversely, the prevalence was low in the liver (0.71%, N = 140) and absent in brain samples (0.0%, N = 17), which have previously been shown to be target organs during acute infections. Phylogenetic analysis showed homology between our sequences and those from recent outbreaks in Israel and Thailand. Given that these findings were based on nucleic acid detection by PCR, future studies should seek to isolate the virus from fish in Lake Victoria and show its ability to cause disease and virulence in susceptible fish.     

Dietary Lactobacillus plantarum supplementation decreases tissue lead accumulation and alleviates lead toxicity in Nile tilapia (Oreochromis niloticus)

Pollution by the heavy metal lead (Pb) has become a threat to both aquaculture and food safety. In this study, the protective effects of a selected probiotic with good Pb binding capacity (Lactobacillus plantarumCCFM8661) against waterborne Pb exposure were evaluated in Nile tilapia (Oreochromis niloticus). One hundred and eighty fish were divided into four groups as control, Pb‐only, Pb‐plus‐CCFM8661 and CCFM8661‐only. Fish were exposed to a dose of waterborne Pb level at 1 mg L^?1 for 4 weeks, and the probiotic was administered at 10⁸ CFU g^?1 in fish diet twice daily. The results showed that dietary supplementation of CCFM8661 ameliorated the growth performance and prevented the death of Pb‐exposed fish. Cotreatment with Pb and CCFM8661 also effectively decreased Pb accumulation in kidney, liver, gonad, brain, gills and muscle. Moreover, the administration of this probiotic alleviated Pb‐induced oxidative stress, recovered digestive enzyme activities and blood δ‐aminolevulinic acid dehydratase activity, reversed alterations in innate immune status and decreased the frequencies of the nuclear abnormalities in peripheral blood erythrocytes of fish. These results suggested that CCFM8661 may be a novel dietary supplement against Pb toxicity at least in tilapia.     

Variability of the nitrifying bacteria in the biofilm and water column of a recirculating aquaculture system for tilapia (Oreochromis niloticus) production

The aim of this study was to evaluate variability of nitrifying bacterial community in the biofilm and in the water of a recirculating aquaculture systems (RAS) in a tilapia farming in order to determine if nitrification process is dependent, or not, of nitrifying bacteria abundance. Biofilm and water samples were collected periodically for 30 days and analysed with the fluorescent in situ hybridization (FISH) technique, used to quantify ammonia‐oxidizing bacteria (AOB) and nitrite‐oxidizing bacteria (NOB). Ammonia presented the peak in the first week, while the nitrite's maximum was recorded in the second week. Nitrate increased steadily, indicating nitrification activity. Total bacterial abundance in biofilm increased continuously, while in water, it did not change significantly. In the biofilm, number of AOB was high at beginning, decreased after few days and increased again following augment of ammonia. Number of NOB also showed an increase in abundance in biofilm following the increment of nitrite and nitrate. In water, AOB and NOB did not show major variability. Relative abundance of nitrifying bacteria represented more than 30% of total bacteria in biofilm at beginning of the experiment. Their contribution decreased to >3% in last days. It indicates that nitrifying bacteria are biofilm colonizers, and that their activity seems to be directly related to the concentration of nitrogen compounds. However, contribution of nitrifying bacteria did not vary much along the time. We may conclude that the biofilm‐nitrifying bacteria plays major role in nitrification process in RAS and that the activity of these organisms is dependent of their abundance in response to the concentration of nitrogen compounds.     

Francisella sp., an emerging pathogen of tilapia, Oreochromis niloticus (L.), in Costa Rica

Francisella sp. is an emergent bacterial pathogen that causes acute to chronic disease in warm and cold water cultured and wild fish species. During the past 3 years, the bacterium has been detected in tilapia, Oreochromis niloticus , cultured in Costa Rica. Infected fish presented non-specific clinical signs, such as erratic swimming, anorexia, anaemia, exophthalmia and high mortality. Upon macroscopic and microscopic examination, several internal organs (mainly spleen and kidney) were enlarged and contained white nodules. Histological examination revealed the presence of multifocal granulomatous lesions, with the presence of numerous small, pleomorphic, cocco-bacilli. The bacteria were isolated from infected tilapia on selective media and grown on several media with and without antibiotics. Specific PCR primers to the Francisella genus were used to confirm the preliminary diagnoses. In comparison with several bacterial 16S rRNA sequences, our isolate was found to share 99% identity with other Fransicella spp. isolated from fish, and more than 97% identity to the human pathogen Francisella tularensis . Koch's postulates were fulfilled after experimental intraperitoneal and gill exposure challenges.     

The synthetic antioxidant, ethoxyquin, adversely affects immunity in tilapia (Oreochromis niloticus)

Ethoxyquin (EQ) has been used as an antioxidant in livestock, aquaculture and pet foods. Animal food safety law has established the upper limit of EQ in animal feed at 150 mg kg⁻¹. However, the risk of EQ at the approved level for aquaculture feed (150 mg kg⁻¹) to fish health is unknown. Here, we examine the effect of EQ on the immunity of tilapia ( Oreochromis niloticus ). EQ concentration in the blood reached 0.16 mg L⁻¹ in fish fed EQ at the approved level. This level of EQ inhibited phagocytic activity of leucocytes in vitro and antibacterial activity of whole blood in vivo . Furthermore, pyknosis in the liver was observed throughout the duration of feeding. However, after 30 days of experimental challenge with feed containing 150 mg kg⁻¹ of EQ, no significant difference was observed in mortality. Although EQ at the approved level in feed causes immunosuppression in fish, the severity of immunosuppression does not lead to a lowering of disease resistance for short feeding periods.     

Faecal coliforms in pond water, sediments and hybrid tilapia Oreochromis niloticus×Oreochromis aureus in Saudi Arabia

Total bacterial load, total coliforms faecal coliforms in pond water, sediment, intestine of hybrid tilapia Oreochromis niloticus×Oreochromis aureus and pigeon Columba livia faeces were investigated monthly over a period of 1 year from July 1999 to June 2000. Fish were collected randomly by a cast net. Samples were analysed for coliforms using the multiple‐tube fermentation technique. Results showed total viable bacterial counts in the pond water, sediment, intestine of tilapia and pigeon faeces ranging from 1.8±0.9×10² to 6.0±1.2×10⁴ cfu mL^?1, 3.2±1.2×10⁵ to 2.8±1.5×10⁷ cfu g^?1, 8.2±1.6×10⁵ to 9.9±1.5×10⁷ cfu g^?11.0±0.4×10⁷to9.7±0.2×10⁹ cfu g^?1respectively. The most probable number (MPN) of coliforms and faecal coliforms ranged from 287±12 to ≥1600±0 100 mL^?1 in pond water; the MPN ranges for sediment, tilapia intestine and pigeon faeces were 257±29 to ≥1100±0 g^?1, 237±46 to ≥1100±0 g^?1 and 403±98 to ≥1100±0 g^?1 respectively. The abundance of normal bacteria coliforms was greater in the warm months than in the cold months. Ground water was free from any sort of coliform organisms, and there were no sources of human faecal matter in the pond. So, it is clear that faecal coliforms from pigeon faeces significantly contaminated (P<0.05) the ponds and tilapia intestines. Escherichia coli was the only coliform organism found in pond water, sediment, intestine of tilapia and pigeon faeces.     

Lead (Pb) accumulation,oxidative stress and DNA damage induced by dietary Pb in tilapia (Oreochromis niloticus)

With the increasing occurrence of dietary lead (Pb) contamination in aquaculture, a better understanding of the toxic effects of dietary Pb on aquatic animals is needed. Tilapia (Oreochromis niloticus) were exposed to dietary Pb at concentrations of 0, 100, 400 and 800 μg g^?1 dry weight for 60 days, and Pb accumulation in tissues and blood, oxidative stress in posterior kidney and DNA damage in peripheral blood cells were investigated. The results showed that dietary Pb exposure resulted in significant Pb accumulation in tissues and blood, which increased with the dietary Pb concentrations. Pb accumulated in sampled tissues in the following order: posterior kidney>bone>liver>gill>spleen>testis>muscle>brain. Dietary Pb caused a significant increase in the malondialdehyde level and a significant decrease in the total antioxidant capacity content when compared with the control group (P<0.05), accompanied by concentration‐dependent decreases in the glutathione content, glutathione peroxidase and superoxide dismutase activities. Pb dose‐dependent DNA damage was observed in peripheral blood cells of tilapia exposed to dietary Pb. The results suggest that dietary Pb exposure can induce significant Pb accumulation in tissues and blood, followed by oxidative stress in posterior kidney and DNA damage in peripheral blood cells of tilapia.     

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.     

Depletion of stored nutrients during fasting in Nile tilapia (Oreochromis niloticus) juveniles

We evaluated growth performance and metabolic responses in Nile tilapia (Oreochromic niloticus) juveniles (30.2 ± 0.9 g) subjected to 1 (F1), 2 (F2), or 3 weeks (F3) of fasting and then refed for 10 weeks (10WR) compared to controls (FC), which were fed for the full 13-week trial. Weight gain and specific growth rate (SGR) during fasting were lower in all treatments compared to the FC. However, during refeeding, feed intake/body mass and SGR increased in F1, F2, and F3, inducing partial compensatory growth. The hepatosomatic index (HSI), visceral fat index (VFI), liver glycogen (LG), and carcass lipid levels dropped in all fasted fish compared to FC (P < 0.05), showing a depletion of stored nutrients such as fat and LG. Along with LG, fat reserves were mobilized during fasting to maintain basal metabolism and survival, but these energy constituents returned to control levels at 10WR, at which time HSI was higher in all refed fish compared to FC. Additionally, the variables VFI, LG, and lipid in carcass increased in all refed fish, equaling those of FC at 10WR. The results showed that, in contrast with other protocols that used smaller tilapia juveniles, the feeding strategies utilized for Nile tilapia juveniles in this study (1 to 3 weeks of fasting and 10WR) were able to induce only partial compensatory growth. It can be concluded that in situations that require complete food restriction in juvenile Nile tilapia (30 g), an acceptable strategy is to limit the period of fasting to 1 week or less to minimize losses and to achieve partial compensatory growth.     

Effects of Bacillus subtilis strains and the prebiotic Previda® on growth,immune parameters and susceptibility to Aeromonas hydrophila infection in Nile tilapia,Oreochromis niloticus

This study was conducted to evaluate the individual and combined effects of long‐term feeding of diets containing two probiotic Bacillus subtilis group strains (Aqua NZ and AP193) and the prebiotic Previda^®, a commercial hemicellulose extract, on growth performance, immune parameters and Aeromonas hydrophila susceptibility of juvenile Nile tilapia, Oreochromis niloticus. Nile tilapia of average weight 7.47 ± 0.11 g were fed diets formulated with the probiotics and/or the prebiotic, or a control diet for 8 weeks and, subsequently, challenged with A. hydrophila by intragastric gavage at a dosage of 3.9 × 10⁷ CFU per fish. Fish attained a mean weight of 59.5 ± 0.99 g at the end of the growth period. Under the conditions of the present trial, none of the diets significantly improved mean per cent weight gain (P = 0.70), thermal growth coefficient (P = 0.88) or feed conversion ratio (P = 0.87) of Nile tilapia. Except for the diet containing the prebiotic Previda^® only (P = 0.17), all other diets resulted in significantly higher fish survival compared to the control (P < 0.05). The combined effect of the prebiotic and probiotic strains emerged as the most important diet with respect to mortality reduction. The mean lysozyme and respiratory burst activities did not show any significant differences between treatments and control (P = 0.14 and 0.32, respectively). Thus, these probiotic strains used in this study have the potential to prevent disease due to A. hydrophila in O. niloticus.     

Osmoregulatory actions of growth hormone in juvenile tilapia (Oreochromis niloticus)

Growth hormone (GH) effectively promotes seawater (SW) adaptation in salmonids, but little is known of its effect in tilapias. Experiments were performed to investigate the effects of recombinant eel GH (reGH) on osmoregulatory actions and ultrastructural features of gill chloride cells in juvenile tilapia, Oreochromis niloticus. Tilapia showed a markedly improved SW survival, when directly transferred from freshwater (FW) to 62.5% SW 24h after a single reGH injection (0.25 or 2.5 µg g^-1) or 3 reGH injections (0.25 µg g^-1 every other day). Plasma Na+ and Mg²⁺ levels were significantly reduced by reGH (0.25 and 2.5 µg g^-1) compared with saline injections; Ca²⁺ concentrations were reduced significantly by high dose of reGH (2.5 µg g^-1) after SW transfer. However, fish failed to survive more than 24h when directly transferred to 70% SW, although the fish treated with reGH could survive longer than the controls. When examined by electron microscopy, the chloride cells were identified as mitochondrion-rich and an extensive tubular system was induced by GH treatment. The results of the present study suggest that, similar to its effect on salmonids, GH also exerts acute osmoregulatory actions and enhances SW adaptation in juvenile tilapia. GH also stimulates the differentiation of chloride cells toward SW adaptation.     

Nutrient composition and sensory evaluation of fillets from wild-type Nile tilapia (Oreochromis niloticus, Linnaeus) and a red hybrid (Florida red tilapia × red O. niloticus)

A comparison of composition and sensory evaluation were performed on fillets of Oreochromis niloticus (wild type) and red hybrid tilapia (RHT) (Florida red tilapia ♂× red O. niloticus♀). Fifty male tilapia, 25 O. niloticus (initial weight 159.3 g) and 25 red hybrid (initial weight 132.4 g), were placed in each of three 2.0 m³ volume tanks. The fish were fed a commercial feed containing 35.9% crude protein during the 120‐day experimental period and then killed in cold water (3°C). All the fish were filleted. Twelve fillets from each treatment were used for proximate analyses, five fillets from each treatment were used for fatty acids analyses and the remaining fillets were used for sensory evaluation. The compositional analyses showed similar moisture, true protein and ash content in both genetic groups, but a lower crude fat content was measured in the red hybrid fillets (0.70%) compared with that of O. niloticus fillets (0.97%). No differences between O. niloticus and the red hybrid were observed in their fillet profile of fatty acids. In the sensory evaluation test, a difference in fillet flavour between both genetic groups was perceived. Of the 112 consumers in the preference test, 81.2% perceived a difference in fillet flavour between the two tilapias, with a general preference for the red hybrid over the wild‐type O. niloticus. The benefits of cultivating a RHT with a low lipid content are discussed.