Molecular characterization and pathogenicity of Francisella noatunensis subsp. orientalis isolated from cultured tilapia (Oreochromis sp.) in Taiwan

Francisella noatunensis subsp. orientalis is a causative agent of systemic granulomatous disease in tilapia. The present study was designed to understand the genetic and phenotypic diversities among Taiwanese Fno isolates obtained from tilapia (n = 17) and green Texas cichlid (Herichthys cyanoguttatus) (n = 1). The enzymatic profiles of the isolates were studied using the API ZYM system. Phylogenetic tree analysis of the 16S rRNA and housekeeping gene and pulsed‐field gel electrophoresis (PFGE) were carried out to determine the genotypic characters of all isolates. The phylogenetic tree showed similarity of 99%–100% nucleotide sequences of 16S rRNA and housekeeping genes compared to the Fno references genes from GenBank database. Comparatively, the results revealed an identical profile of enzymatic and PFGE pattern which was distincted from that of F. philomiragia. To understand the pathogenicity, the isolates were intraperitoneal injected to tilapia the gross lesions were observed concomitant with natural outbreak. Median lethal dose upon Nile tilapia and red tilapia were 9.06 × 10³ CFU/fish and 2.08 × 10² CFU/fish, respectively. Thus, our data provide understanding the epidemiology of Taiwanese Fno isolates, and help in development of future control and prevention.     

Depletion of florfenicol amine in tilapia (Oreochromis sp.) maintained in a recirculating aquaculture system following Aquaflor®‐medicated feed therapy

Aquaflor^® [50% w w^?1 florfenicol (FFC)], is approved for use in freshwater‐reared warmwater finfish which include tilapia Oreochromis spp. in the United States to control mortality from Streptococcus iniae. The depletion of florfenicol amine (FFA), the marker residue of FFC, was evaluated after feeding FFC‐medicated feed to deliver a nominal 20 mg FFC kg^?1 BW d^?1 dose (1.33× the label use of 15 mg FFC kg^?1 BW d^?1) to Nile tilapia O. niloticus and hybrid tilapia O. niloticus × O. aureus held in a recirculating aquaculture system (RAS) at production‐scale holding densities. Florfenicol amine concentrations were determined in fillets taken from 10 fish before dosing and from 20 fish at nine time points after dosing (from 1 to 240 h post‐dosing). Water samples were assayed for FFC before, during and after the dosing period. Parameters monitored included daily feed consumption and biofilter function (levels of ammonia, nitrite and nitrate). Mean fillet FFA concentration decreased from 13.77 μg g^?1 at 1‐h post dosing to 0.39 μg g^?1 at 240‐h post dosing. Water FFC concentration decreased from a maximum of 1400 ng mL^?1 at 1 day post‐dosing to 847 ng mL^?1 at 240 h post‐dosing. There were no adverse effects noted on fish, feed consumption or biofilter function associated with FFC‐medicated feed administration to tilapia.     

Dietary taurine improves reproductive performance of Nile tilapia (Oreochromis niloticus) broodstock

This study was conducted to evaluate the effects of supplemental taurine on reproductive performance of Nile tilapia (Oreochromis niloticus) broodstock fed soybean meal‐based diets. Four isonitrogenous (350 g kg^?1 protein), isocaloric (18 MJ kg^?1) diets were formulated to contain 0, 5, 10 and 15 g kg^?1 taurine. The diets were fed to triplicate groups of juvenile Nile tilapia (10–15 g average body weight) at a female: male ratio of 3 : 1, to apparent satiation, three times per day for 130 days. The size at first maturation decreased with increasing dietary taurine to 10 g kg^?1 and levelled off with further taurine supplementation. The time to first spawning was also significantly shorter at 10 g kg^?1 taurine level. Spawning performances, including spawning frequencies, total number of spawnings per tank, number of spawnings per female and absolute fecundity, were all significantly improved with increasing dietary taurine up to 10 g kg^?1. However, the quadratic regression analyses indicated that the maximum spawning performance occurred at 8 g kg^?1 of supplemental taurine. Eggs produced from broodstock fed 10 g kg^?1 taurine exhibited significantly higher hatchability and required shorter time for hatching and yolk‐sac absorption and also resulted in higher larval weight than at other dietary taurine levels. The highest egg protein, total amino acids and taurine were also obtained at 10 g kg^?1 taurine. These results suggest that 8 g kg^?1 dietary taurine is required for optimum reproductive outputs of Nile tilapia broodstock.     

Application of multiplex quantitative polymerase chain reaction methods to detect common bacterial fish pathogens in Nile tilapia,Oreochromis niloticus,hatcheries in Costa Rica

Edwardsiella spp., Streptococcus spp., and Francisella noatunensis subsp. orientalis are some of the most important fish pathogens affecting global tilapia, Oreochromis spp., aquaculture. In Costa Rica, the aquaculture industry is dominated by freshwater‐cultured Nile tilapia, Oreochromis niloticus, which are raised in all seven national provinces. At present, little is known regarding the diversity of pathogens present in these facilities, and definitive identification of agents associated with disease outbreaks are rare. To evaluate the prevalence of common bacterial pathogens in these systems, this study used multiplex quantitative polymerase chain reaction (qPCR) assays targeting Edwardsiella, Streptococcus, and Francisella species as a diagnostic and surveillance tool. In 2017, seven different tilapia hatcheries were visited, and 350 fingerlings were subjected to necropsy and molecular diagnostic evaluation. Fish exhibiting gross signs of disease were subjected to histological and microbiological analysis. For the first time, Edwardsiella anguillarum was recovered and molecularly confirmed from diseased tilapia in Costa Rica. In addition, F. noatunensis subsp. orientalis was identified in a region of Costa Rica where it had not been previously reported.     

Effectiveness of eugenol sedation to reduce the metabolic rates of cool and warm water fish at high loading densities

Effects of eugenol (AQUI‐S^®20E, 10% active eugenol) sedation on cool water, yellow perch Perca flavescens (Mitchill), and warm water, Nile tilapia Oreochromis niloticus L. fish metabolic rates were assessed. Both species were exposed to 0, 10, 20 and 30 mg L^?1 eugenol using static respirometry. In 17°C water and loading densities of 60, 120 and 240 g L^?1, yellow perch controls (0 mg L^?1 eugenol) had metabolic rates of 329.6–400.0 mg O₂ kg^?1 h^?1, while yellow perch exposed to 20 and 30 mg L^?1 eugenol had significantly reduced metabolic rates of 258.4–325.6 and 189.1–271.0 mg O₂ kg^?1 h^?1 respectively. Nile tilapia exposed to 30 mg L^?1 eugenol had a significantly reduced metabolic rate (424.5 ± 42.3 mg O₂ kg^?1 h^?1) relative to the 0 mg L^?1 eugenol control (546.6 ± 53.5 mg O₂ kg^?1 h^?1) at a loading density of 120 g L^?1 in 22°C water. No significant differences in metabolic rates for Nile tilapia were found at 240 or 360 g L^?1 loading densities when exposed to eugenol. Results suggest that eugenol sedation may benefit yellow perch welfare at high densities (e.g. live transport) due to a reduction in metabolic rates, while further research is needed to assess the benefits of eugenol sedation on Nile tilapia at high loading densities.     

Assessment of Aquaflor(®) , copper sulphate and potassium permanganate for control of Aeromonas hydrophila and Flavobacterium columnare infection in sunshine bass, Morone chrysops female × Morone saxatilis male

Two experiments were conducted to test the effectiveness of different therapeutants against a mixed infection of Aeromonas hydrophila and Flavobacterium columnare in sunshine bass. Experiment 1 evaluated copper sulphate, florfenicol‐medicated feed and potassium permanganate (KMnO₄) against a natural mixed infection. Experiment 2 further evaluated copper sulphate as a treatment to control an experimental mixed infection. In experiment 1, naturally infected untreated fish had the lowest final survival per cent, at 71%, while florfenicol‐medicated feed at 15 mg kg^?1 body weight for 10 days or copper sulphate at 2.1 mg L^?1 (1% of the total alkalinity) for 24 h produced the highest final survivals, at 90% and 88%, respectively. The final survival of the naturally infected fish administered florfenicol‐medicated feed was significantly different (P < 0.1) from the untreated fish. The survival curves for the florfenicol and the copper sulphate at 2.1 mg L^?1 were significantly improved from the untreated fish. In experiment 2, fish were challenged by waterborne exposure to A. hydrophila and F. columnare and either not treated or treated with copper sulphate at 2.1 mg L^?1. At the end of experiment 2, the per cent survival of the challenged fish treated with copper sulphate (99%) was significantly higher (P < 0.05) than the non‐treated (61%). The results illustrate clear benefit of florfenicol and copper sulphate against a mixed infection of A. hydrophila and F. columnare.     

Whole cell inactivated autogenous vaccine effectively protects red Nile tilapia (Oreochromis niloticus) against francisellosis via intraperitoneal injection

Francisella noatunensis subsp. orientalis is a pathogen of tilapia and other warm‐water fish for which no vaccines are commercially available. In this study, a whole cell formalin‐inactivated vaccine was developed for the first time using the highly virulent isolate STIR‐GUS‐F2f7 and the oil‐based adjuvant Montanide? ISA 763A VG. The efficacy of the vaccine was assessed in red Nile tilapia via intraperitoneal (i.p.) injection using homologous experimental infection and correlates of protection such as seral antibody production and bacterial loads in the spleen. For immunization, fish were i.p. injected with 0.1 ml of the vaccine, the adjuvant alone or PBS. At 840 degree days post‐vaccination, all fish were i.p. injected with 4.0 × 10³ CFU/fish of pathogenic bacteria. The RPS at the end of the trial was 100% in the vaccinated group with significantly higher survival than in the adjuvant and control groups. The RPS in the adjuvant group was 42%, and no significant difference was seen in survival between this and the PBS group. Moreover, significantly higher antibody titres in the serum and significantly lower bacterial loads in the spleen were detected in the vaccinated fish by ELISA and qPCR, respectively. These findings highlight the potential of autogenous vaccines for controlling francisellosis in tilapia.     

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.     

The effect of dietary methionine concentrations on growth performance of juvenile Nile tilapia (Oreochromis niloticus) fed diets with two different digestible energy levels

A growth trial was conducted to examine the effect of dietary digestible energy (DE) content on methionine (Met) utilization and requirement in juvenile Nile tilapia (Oreochromis niloticus). Ten iso‐nitrogenous (288 g kg^?1 protein) practical diets, with two DE levels (10.9 MJ kg^?1; 12.4 MJ kg^?1) and five methionine supplementation levels (0, 1, 2, 4 and 6 g kg^?1), were hand‐fed twice daily to triplicate groups of Nile tilapia (initial body weight 8.95 ± 0.06 g) for 8 weeks. Weight gain (WG) and specific growth rate (SGR) increased significantly with increasing dietary methionine concentration at the same DE content (P < 0.001). At the same dietary methionine level, WG and SGR of fish fed high‐DE diets were significantly higher than that of fish fed low‐DE diets (P = 0.0001), although no interaction was found between dietary DE and methionine supplementation. Based on quadratic regression analysis between dietary methionine concentration and weight gain, optimal methionine requirement for maximum growth, expressed as g Met required kg^?1 diet (low‐ versus high‐DE diets), increased as diet DE concentration increased (7.34 versus 9.90 g kg^?1 diet, respectively; with cysteine 4.70 g kg^?1 diet). The results indicated that diet DE content affects methionine utilization and requirement in juvenile Nile tilapia, fish fed high‐DE diets required more methionine for maximum growth.     

Yeast extract on growth,nutrient utilization and haemato‐immunological responses of Nile tilapia

Dietary supplementation (0, 10, 20, 40, 60, 80 g kg^?1) of a nucleotide source (NuPro^®) was evaluated on growth, nutrient utilization, haemato‐immunological responses and resistance to experimental infection of Nile tilapia. Fish (2.63 ± 0.63 g) were fed experimental diets to satiation three times a day for 75 days. Feed intake increased linearly with increasing levels of NuPro^® promoting higher weight gain (P < 0.05). Survival, feed conversion and protein retention were not affected. Following the growth trial, fish were challenged with Aeromonas hydrophila by intraperitoneal injection. Survival and innate immune responses (phagocytosis and lysozyme activity) were not significantly affected. However, among the haematological variables, thrombocyte, leucocyte and monocyte counts increased linearly (P < 0.05). The indirect measurement of digestibility in juvenile Nile tilapia (122.32 ± 11 g) indicated a linear decrease in protein (1.41%), dry matter (5.72%) and energy (4.10%) digestibility coefficients as dietary supplementation increased (P < 0.05). NuPro^® showed to be a beneficial additive to be supplemented in diets for juvenile Nile tilapia, providing 32.8% increase in feed intake and 28.8% in weight gain, as well as in thrombocyte, leucocyte and monocyte counts in blood, possibly indicating a more efficient defence response in the highest dietary supplementation levels tested.     

Effects of dietary arginine on growth performance,feed utilization,haematological parameters and non‐specific immune responses of juvenile Nile tilapia (Oreochromis niloticus L.)

An 8‐week feeding trial was conducted to evaluate the effects of dietary arginine on growth performance, feed utilization, haematological parameters and non‐specific immune responses of juvenile Nile tilapia (6.03 g). Five isonitrogenous and isoenergetic diets were formulated to contain graded levels of L‐arginine (8.5 g kg^?1, 11.5 kg^?1, 15.3 kg^?1, 18.8 kg^?1 and 22.4 kg^?1 dry diet) from dietary ingredients and crystalline arginine. Each diet was randomly assigned to triplicate groups of 25 juvenile fish (6.04 ± 0.02 g) three times daily (8:30, 12:30, and 17:00 hours) to apparent satiation. Results showed that the weight gain (WG) and special growth rate (SGR) increased with increasing dietary arginine levels up to 15.3 kg^?1 and remained nearly the same thereafter. Arginine supplementation had no impact on the survival, body composition and haematological parameters of tilapia. However, the immune responses (plasma nitric oxide content, total nitric oxide synthase (T‐NOS) and lysozyme activity) significantly (P < 0.05) increased with increasing dietary argentine level after stress. Quadratic regression analysis (y = ?0.7147x² + 25.986x + 147.53 R² = 0.8736) on weight gain against dietary arginine levels indicated that the optimal dietary arginine requirement was estimated to be 18.2 kg^?1 of the diet (corresponding to 6.24% of dietary protein on a dry weight basis). The present study indicates that arginine has an immunostimulatory potential to help juvenile Nile tilapia cope with stress conditions such as Streptococcus iniae infection.     

Dietary taurine enhances growth and feed utilization in larval Nile tilapia (Oreochromis niloticus) fed soybean meal‐based diets

This study was conducted to evaluate the effects of dietary taurine on growth performance and feed utilization of Nile tilapia (Oreochromis niloticus) larvae. Four plant protein‐based, isonitrogenous (400 g kg^?1 protein), isoenergetic (19 MJ kg^?1) diets supplemented with four taurine concentrations (0.0, 5.0, 10.0 and 15.0 g kg^?1; designated as T₀, T_0.5, T₁ and T_1.5, respectively) were prepared. The diets were fed to triplicate groups of fish larvae (0.024 g average body weight), to apparent satiation, three times per day for 60 days. Larval growth rates and feed utilization efficiency were significantly improved with increasing supplemental taurine up to 10 g kg^?1 and decreased with further taurine supplementation. The quadratic regression analyses indicated that the maximum larval performance occurred at about 9.7 g kg^?1 of total dietary taurine. Fish survival was significantly lower at 15 g kg^?1 dietary taurine than at other taurine levels. Body protein significantly increased, while body moisture and ash decreased, with increasing dietary taurine up to 10 g kg^?1 and decreased with further taurine supplementation to 15 g kg^?1. Body lipid was not significantly affected by dietary taurine concentration. A number of body amino acids (tryptophan, arginine, histidine, leucine, isoleucine, valine, alanine, glycine, threonine and taurine) significantly increased with increasing supplemental taurine up to 10 g kg^?1 and then decreased with further increase in dietary taurine levels. The rest of body amino acids were not significantly affected by dietary taurine. The present results suggest that about 9.7 g kg^?1 dietary taurine is required for optimum performance of Nile tilapia larvae fed soybean meal‐based diets.     

Sensitivity and specificity of real‐time PCR and bacteriological culture for francisellosis in farm‐raised Nile tilapia (Oreochromis niloticus L.)

Despite the worldwide occurrence of Francisella noatunensis subsp. orientalis (Fno) infection in farmed tilapia, sensitivity and specificity estimates of commonly used diagnostic tests have not been reported. This study aimed to estimate the sensitivity and specificity of bacteriological culture and qPCR to detect Fno infection. We tested 559 fish, sampled from four farms with different epidemiological scenarios: (i) healthy fish in a hatchery free of Fno; (ii) targeted sampling of diseased fish with suggestive external clinical signs of francisellosis during an outbreak; (iii) convenience sampling of diseased and clinically healthy fish during an outbreak; and (iv) sampling of healthy fish in a cage farm without a history of outbreaks, but with francisellosis reported in other farms in the same reservoir. The qPCR had higher median sensitivity (range, 48.8–99.5%) than culture (range, 1.6–74.4%). Culture had a substantially lower median sensitivity (1.6%) than qPCR (48.8%) to detect Fno in carrier tilapia (farm 4). Median specificity estimates for both tests were >99.2%. The qPCR is the superior test for use in surveillance and monitoring programmes for francisellosis in farmed Nile tilapia, but both tests have high sensitivity and specificity which make them fit for use in the diagnosis of Fno outbreaks.     

Improvement of growth and antioxidant status in Nile tilapia,Oreochromis niloticus,fed diets supplemented with mushroom stalk waste hot water extract

Mushroom stalk waste (MSW) generated from the oyster mushroom industry is drastically increasing and safe disposal of MSW is becoming a critical issue worldwide. Mushrooms are an established, excellent source of nutraceuticals and antioxidants, and therefore MSW may confer similar effects when consumed. The severe deterioration of aquatic habitat may compromise the homoeostasis of antioxidative metabolism of Nile tilapia, Oreochromis niloticus, which resulted in a reduced total production. Based on this paradigm, hot water extracts (HWE, 2‐ and 5‐h) of MSW were evaluated as a supplement in fish feed and determined the effect on growth performance and the antioxidant status of Nile tilapia, O. niloticus, in vivo. The HWE 5‐h contained significantly higher amounts of nutrient and β‐glucan than HWE 2‐h (P < 0.05). Four standardized in vitro antioxidant assays showed that HWE 5‐h was a better antioxidant agent than HWE 2‐h (P < 0.05). Tilapia fingerlings fed 5 g kg^?1 HWE supplemented feed produced better (P < 0.05) growth and higher antioxidant enzyme activities than 10 g kg^?1 HWE, CD and BD. Thus, HWE of MSW included at 5 g kg^?1 may be beneficial to tilapia farming and may also provide a way of disposing the waste created by mushroom farming.     

Dietary threonine requirement to optimize protein retention and fillet production of fast‐growing Nile tilapia

Threonine is the third‐limiting essential amino acid in diets based on cereal ingredients. A 4‐week trial was conducted to determine the threonine requirement of large Nile tilapia based on fish growth, feed efficiency, body composition, protein and amino acid retention. Six hundred fish (563.3 ± 15.1 g) were distributed into twenty 1.2‐m³ cages. Five diets containing 288 g kg^?1 of crude protein, 12.7 MJ kg^?1 of digestible energy and 8.9, 10.5, 12.2, 13.7 and 15.4 g kg^?1 of threonine were elaborated. Fish were hand‐fed five times a day to extruded diets. Significantly, differences in growth performance and amino acids retention among the treatments were observed. Fish fed 10.5 g kg^?1 of threonine showed higher daily weight gain, gutted weight and fillet weight (P < 0.05) compared to fish fed with other experimental diets. Diets containing 8.9–15.4 g kg^?1 of threonine did not affect whole body and muscle proximate composition. Based on second‐order regression analysis, the dietary threonine requirement estimated based on final gain, fillet weight and fillet yield was 12, 12.1 and 11.5 g kg^?1 diet, respectively. The dietary threonine requirement for maximum fillet yield of Nile tilapia was estimated to be 11.5 g kg^?1.     

The effect of β‐1,3‐glucan derived from Euglena gracilis (Algamune™) on the innate immunological responses of Nile tilapia (Oreochromis niloticus L.)

Algamune^? is a commercial additive produced from Euglena gracilis, providing a rich source of the β‐1,3‐glucan paramylon. Isolated kidney phagocytes of Nile tilapia were incubated with graded doses (0, 8, 16, 32, 64 and 128 μg/ml) of Algamune^? and purified paramylon to gauge their ability to elicit the production of reactive oxygen species. A linear response was observed for extracellular superoxide anion for both sources but only Algamune^? for intracellular superoxide anion. After corroborating the immunostimulant properties ex vivo, a feeding trial was conducted to evaluate the dietary supplementation of Algamune^? (0, 100, 200, 400 and 800 mg/kg of diet) for Nile tilapia. Fish were fed for 3 weeks, after which, fish were sampled for blood and head kidney phagocytes. The remaining fish were challenged with Streptococcus iniae. Macrophage extracellular superoxide anion production was significantly elevated in fish fed diets with 200 mg of Algamune^? kg^?1 when compared to fish fed the basal diet. Even though the disease challenge did not show statistical differences, it is worth mentioning that fish fed intermediate doses of Algamune^? had lowest numerical mortality values. Therefore, Algamune^? was demonstrated to enhance some immunological responses of tilapia both in ex vivo and in vivo.     

Efficacy of an experimentally inactivated Streptococcus agalactiae vaccine in Nile tilapia (Oreochromis niloticus) reared in Brazil

Tilapia aquaculture is one of the fastest‐growing segments of fish production in Brazil. Nile tilapia (Oreochromis niloticus) is largely cultivated in the state of Parana, where Streptococcus agalactiae is the cause of severe disease outbreaks. The objective of this paper was to evaluate an inactivated S. agalactiae vaccine in tilapia for the control of streptococcal disease outbreaks. Tilapia, weighing approximately 20 g each, were intraperitoneally (i.p.) inoculated with 0.1 mL of the vaccine at a dose of 2.0 × 10⁸ colony‐forming unit (CFU) mL^?1. One group of tilapia (treatment 1) received one vaccine dose, and the other group of tilapia (treatment 2) received two doses, with an interval of 21 days. The control group was i.p. inoculated with 0.1 mL tryptic soy broth fish^?1. Immunized and control tilapia were i.p. challenged with 0.1 mL of 3.0 × 10⁷ CFU mL^?1 at 30 days post vaccination. The fish were monitored daily for disease signs and for mortality for 16 days post challenge. A statistically significant difference (P=0.0045) was found between the mortality of treatments 1 and 2. The value of relative per cent of survival of 83.6% and 96.4%, respectively, indicate that this vaccine was efficient in Nile tilapia.     

Vertical transmission and concurrent infection of multiple bacterial pathogens in naturally infected red tilapia (Oreochromis spp.)

Horizontal transmission of various bacterial pathogens in tilapia is well described, but there is scant information regarding their vertical transmission. This study aimed to determine the possibility of vertical transmission of two commonly reported bacterial pathogens (Francisella noatunensis subsp. orientalis and Shewanella putrefaciens) in natural stocks of red tilapia (Oreochromis spp.). Vertical transmission of these pathogens via gametes was evaluated using in vitro fertilization from 10 different families and analysing for the presence of bacteria in milt, unfertilized eggs, fertilized eggs and offspring at various ages (1‐day‐old larvae, 10‐day‐old fry and 30‐day‐old fingerlings), as well as water samples using colorimetric LAMP assay. The study revealed that both F. n. orientalis (6/10) and S. putrefaciens (4/10) was transmitted vertically to the fertilized eggs. Analysis of the water samples from different water sources (brood stock tanks, hatching chamber and larval rearing tanks) showed that both the pathogens were present in water samples with highest prevalence for F. n. orientalis followed by S. putrefaciens. Analyses for the presence of two pathogens in various organs (gonads, gill, liver, spleen, kidney and brain) of the healthy tilapia broodstock without any clinical symptoms of disease demonstrated they were carriers of S. putrefaciens and F. n. orientalis. This is the first documented evidence that vertical transmission via the broodstock of tilapia may also play an important role in transmitting these problematic pathogens to their progeny and underlines the necessity to modify the current disease management strategies in tilapia aquaculture.     

Growth performance,oxidative stress indices and hepatic carbohydrate metabolic enzymes activities of juvenile Nile tilapia,Oreochromis niloticus L., in response to dietary starch to protein ratios

The effect of various dietary starch to proteins ratios (STA/P) on growth performance, oxidative status and liver enzyme activities involved in intermediary metabolism in juvenile Nile tilapia was evaluated. Four isocaloric‐practical diets (12.73 MJ kg^?1 digestible energy) with increasing STA/CP ratios were formulated. These were designated D₀ (344 g crude protein (CP) and 163.5 g starch (STA) kg^?1), D₁ (310 g CP and 243 g STA kg^?1), D₂ (258 g CP and 322 g STA kg^?1) and D₃ (214 g CP and 401 g STA kg^?1). Each diet was fed to triplicate groups of 60 fish (2.7 g) for 45 days. Compared with the control diet (D₀), significantly (P < 0.05) depressed growth and feed efficiency were observed only in the groups fed on diet D₃. The activities of hepatic enzymes involved in glycolysis and lipogenesis pathways were significantly enhanced in groups fed on diet D₃ compared with other diets. A significant (P < 0.05) increase in catalase activity was detected only in groups fed on diet D₃. Similarly, a significant (P < 0.05) enhancement in superoxyde dismutase, glutathione S‐transferases and glutathione peroxidise was observed in groups fed on diets D₂ and D₃ compared with other diets. Results demonstrate the ability of juvenile Nile tilapia to spare protein by dietary carbohydrate.