Toxicological and hematological effect of <Emphasis Type="Italic">Terminalia arjuna</Emphasis> bark extract on a freshwater catfish, <Emphasis Type="Italic">Heteropneustes fossilis</Emphasis>

Increasing demand for eco-friendly botanical piscicides and pesticides as replacements for harmful synthetic chemicals has led to investigation of new sources of plant materials. Stem bark of Terminalia arjuna, which has been used as a popular folk medicine since ancient time, was examined for its piscicidal activity. This study aims to determine toxicity of ethanol extract of T. arjuna bark on fresh water stinging catfish (Heteropneustes fossilis), along with evaluation of changes in hematological parameters of the fishes exposed to a lethal concentration. The percent mortality of fishes varied significantly in response to concentrations of the extract and exposure times (between exposure time F = 36.57, p < 0.001; between concentrations F = 39.93, p < 0.001). The lethal concentrations (LC₅₀) of ethanol extract were found to be 12.7, 8.94, 5.63 and 4.71 mg/l for 24, 48, 72 and 96 h, respectively. During acute toxicity test, blood samples of treatment fishes showed significant decreases in the red blood cells count, hematocrit content, hemoglobin concentration, mean corpuscular hemoglobin concentration and plasma protein level when compared to those of the control group, while there were significant increases in the mean corpuscular volume, mean corpuscular hemoglobin, white blood cells count and plasma glucose concentration. These results suggest that T. arjuna bark extract could be considered as a potent piscicide due to its toxic effect on fish, particularly fish hematology.     

Changes in the physicochemical and microbiological properties of dried anchovy <Emphasis Type="Italic">Engraulis japonicus</Emphasis> during storage

This study investigated the changes in the pH, thiobarbituric acid (TBA) value, volatile basic nitrogen (VBN), color L*, a*, b*, shear force, and microbial content (total plate count; TPC) of dried anchovy Engraulis japonicus in a home refrigerator and freezer at 2, ??1, ??5, and ??20 °C. The TBA value and shear force significantly decreased with increasing storage duration at all of the tested temperatures (p?<?0.05), whereas the VBN and TPC values increased (VBN significantly so) with increasing storage duration (p?<?0.05). The anchovy L* values decreased, whereas the a* (redness) and b* (yellowness) values increased as the storage period was lengthened. The pH decreased after 9 months. The results suggest that the optimum storage period of the dried anchovy is within 6 months. It can be stored below 2 °C for 3 months, but for storage exceeding 3 months, it should be frozen at ??5 °C.     

<Emphasis Type="Italic">Virgibacillus proomii</Emphasis> and <Emphasis Type="Italic">Bacillus mojavensis</Emphasis> as probiotics in sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) larvae: effects on growth performance and digestive enzyme activities

This study examined the effects of two probiotics (Virgibacillus proomii and Bacillus mojavensis) on the digestive enzyme activity, survival and growth of Dicentrarchus labrax at various ontogenetic stages in three separate experiments. These probiotics were incorporated as single or mixed into fish feed for a period of 60 days. The growth parameters, proximate composition of whole body, digestive enzymes and gut microbiology were monitored at regular. The increments in length and weight and the survival were significantly higher (P < 0.05), and the values of food conversions were significantly lower (P < 0.05) in fishes fed the probiotic. The administration of V. proomii and B. mojavensis in diet resulted in an increase (P > 0.05) in body ash and protein content and in the specific activity of phosphatase alkaline and amylase in the digestive tract of all the fishes. V. proomii and B. mojavensis persisted in the fish intestine and in the feed in high numbers during the feeding period (group 1: 5.8 × 10⁴ CFU/ml, group 2: 9.6 × 10⁴ CFU/ml, and group 3: 9.8 × 10⁴ CFU/ml day 60). The two probiotics V. proomii and B. mojavensis were adequate for improved growth performance and survival and for healthy gut microenvironment of the host.     

Molecular mobility of fish flesh measured by low-field nuclear magnetic resonance (LF-NMR) relaxation: effects of freeze–thaw cycles

In this study, the molecular mobility of fish flesh was measured by low field nuclear magnetic resonance (LF-NMR) relaxation. Sardine, tuna and mackerel were frozen at ?40 °C and stored for 1 day (24 h); and then these samples were thawed at room temperature (20 °C). The relaxation of water protons in fish flesh was measured for fresh (i.e., before freezing) and multi-cycle freeze–thaw samples (i.e., up to 12 times). Three domains from different pools of protons (i.e., low-mobile, medium-mobile and high-mobile) were identified from the relaxation curve. The T _2b (low-mobile), T ₂₁ (medium-mobile) and T ₂₂ (high-mobile) indicated the proton populations in the protein molecules, strongly bound water molecules, and weakly bound water molecules, respectively. In all cases, the relaxation time (T _2b: sardine r = 0.736 and p < 0.01, tuna r = 0.857 and p < 0.001, mackerel r = 0.904 and p < 0.001; and T ₂₂: sardine r = 0.956 and p < 0.0001, tuna r = 0.927 and p < 0.0001, mackerel r = 0.890 and p < 0.0001) increased with the freeze–thaw cycles and it reached a nearly constant value after 6 freeze–thaw cycles. The increased relaxation time (i.e., higher mobility) up to 6 freeze–thaw cycles could be due to the increase in proton mobility. However, relaxation time (T ₂₁: sardine r = ?0.510 and p > 0.05, tuna r = 0.162 and p > 0.5, mackerel r = 0.513 and p > 0.01) showed insignificant change with the increase of freeze–thaw cycles, which indicated minimal change in the medium-mobile protons. The results in this study revealed that the changes in proton mobility in the fish flesh during freeze–thaw cycles could be identified using T _2b and T ₂₂ relaxation of LF-NMR.     

The effects of starvation on fast-start escape and constant acceleration swimming performance in rose bitterling (<Emphasis Type="Italic">Rhodeus ocellatus</Emphasis>) at two acclimation temperatures

To investigate the effects of starvation and acclimation temperature on the escape ability of juvenile rose bitterling (Rhodeus ocellatus), we measured the fast-start escape and constant acceleration swimming performance of fish fasted for 0 (control), 1 and 2 weeks and half-lethal periods (6 or 4 weeks) at two temperatures (15 and 25 °C). Fish acclimated at a high temperature exhibited shorter response latency (R), higher maximum linear velocity (V _max) and longer escape distance during escape movement (D _120ms) than those at the low temperature. Starvation resulted in a significant decrease in V _max and D _120ms at either low or high temperature and a significant increase in R at only the high temperature in the half-lethal period groups (P < 0.05). The relationship between V _max (Y, m s^?1) and starvation time (X, week) was Y ₁₅ = ?0.062X + 1.568 (r = ?0.665, n = 36, P < 0.001) at low temperature and Y ₂₅ = ?0.091X + 1.755 (r = ?0.391, n = 40, P = 0.013) at high temperature. The relationship between U _cat (Y, cm s^?1) and starvation time (X, week) was Y ₁₅ = ?1.649X + 55.418 (r = ?0.398, n = 34, P = 0.020) at low temperature and Y ₂₅ = ?4.917X + 62.916 (r = ?0.793, n = 33, P < 0.001) at high temperature. The slopes of equations showed a significant difference between low and high temperature (F _1,63 = 9.688, P = 0.003), which may be due to the different energy substrate utilization when faced with food deprivation at different temperatures.     

Molecular cloning and expression analysis of <Emphasis Type="Italic">Megalobrama amblycephala</Emphasis> transferrin gene and effects of exposure to iron and infection with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

Transferrin (Tf) plays an important function in iron homeostasis and metabolism of organisms. In this study, we identified and characterized the Tf gene in Megalobrama amblycephala and evaluated its expression in basal conditions as well as after iron overload and experimental infection with Aeromonas hydrophila. Furthermore, we studied the iron binding properties of recombinant Tf. The full-length M. amblycephala Tf complementary DNA (cDNA) (GenBank accession no.: KX698308) of 2245 bp was cloned and contained a 1953 bp open reading frame (ORF) encoding 650 amino acid residues and flanked by a 68 bp 5′ and a 204 bp 3′ untranslated regions (UTR). Predicted conservative structure illustrated that M. amblycephala Tf consisted of two conservative Tf domains. Amino acid sequence alignment revealed that M. amblycephala Tf had high similarity with that of cyprinids deposited in Genbank, and phylogenetic analysis showed that M. amblycephala Tf clustered with Ctenopharyngodon idella and Hypophthalmichthys molitrix. Tissue expression pattern analyses demonstrated that the liver was the main Tf mRNA expressing organ, being significantly higher than other tissues (p < 0.05). In the liver, Tf mRNA expression in fish artificially injected with the pathogenic bacteria A. hydrophila was significantly upregulated, reaching a peak at 12 h post injection (hpi) and then decreasing afterward. The expression in FeCl₃-injected fish showed a similar tendency, but reached a peak at 8 hpi. Meanwhile, fish serum iron significantly decreased following A. hydrophila injection, but increased to peak at 4 hpi and then decreased in FeCl₃-injected fish. The recombinant M. amblycephala Tf showed iron binding capacity using CAS analysis. These results are helpful to understand the structure and regulation of expression of Tf, as well as the specific function of Tf for both immune responses and iron homeostasis.     

Dietary supplementation of curcumin augments heat stress tolerance through upregulation of <Emphasis Type="Italic">nrf-2</Emphasis>-mediated antioxidative enzymes and <Emphasis Type="Italic">hsps</Emphasis> in <Emphasis Type="Italic">Puntius sophore</Emphasis>

Heat stress is one of the major environmental concerns in global warming regime and rising temperature has resulted in mass mortalities of animals including fishes. Therefore, strategies for high temperature stress tolerance and ameliorating the effects of heat stress are being looked for. In an earlier study, we reported that Nrf-2 (nuclear factor E2-related factor 2) mediated upregulation of antioxidative enzymes and heat shock proteins (Hsps) provide survivability to fish under heat stress. In this study, we have evaluated the ameliorative potential of dietary curcumin, a potential Nrf-2 inducer in heat stressed cyprinid Puntius sophore. Fishes were fed with diet supplemented with 0.5, 1.0, and 1.5% curcumin at the rate 2% of body weight daily in three separate groups (n = 40 in each group) for 60 days. Fishes fed with basal diet (without curcumin) served as the control (n = 40). Critical thermal maxima (CTmax) was determined for all the groups (n = 10, in duplicates) after the feeding trial. Significant increase in the CTmax was observed in the group fed with 1.5% curcumin- supplemented fishes whereas it remained similar in groups fed with 0.5%, and 1% curcumin-supplemented diet, as compared to control. To understand the molecular mechanism of elevated thermotolerance in the 1.5% curcumin supplemented group, fishes were given a sub-lethal heat shock treatment (36 °C) for 6 h and expression analysis of nrf-2, keap-1, sod, catalase, gpx, and hsp27, hsp60, hsp70, hsp90, and hsp110 was carried out using RT-PCR. In the gill, expression of nrf-2, sod, catalase, gpx, and hsp60, hsp70, hsp90, and hsp110 was found to be elevated in the 1.5% curcumin-fed heat-shocked group compared to control and the basal diet-fed, heat-shocked fishes. Similarly, in the liver, upregulation in expression of nrf-2, sod, catalase, and hsp70 and hsp110 was observed in 1.5% curcumin supplemented and heat shocked group. Thus, this study showed that supplementation of curcumin augments tolerance to high temperature stress in P. sophore that could be attributed to nrf-2-induced upregulation of antioxidative enzymes sod, catalase, gpx, and the hsps.     

Pre-sedation and transport of <Emphasis Type="Italic">Rhamdia quelen</Emphasis> in water containing essential oil of <Emphasis Type="Italic">Lippia alba</Emphasis>: metabolic and physiological responses

The effects of transporting silver catfish (Rhamdia quelen) for 6 h in plastic bags containing 0 (control), 30 or 40 µL/L of essential oil (EO) from Lippia alba leaves were investigated. Prior to transport, the fish in the two experimental groups were sedated with 200 µL/L of EO for 3 min. After transport, dissolved oxygen, carbon dioxide, alkalinity, water hardness, pH, temperature and un-ionized ammonia levels in the transport water did not differ significantly among the groups. However, total ammonia nitrogen levels and net Na⁺, Cl^? and K⁺ effluxes were significantly lower in the groups transported with EO of L. alba than those in the control group. PvO₂, PvCO₂ and HCO₃ ^? were higher after transporting fish in 40 µL/L of EO of L. alba, but there were no significant differences between groups regarding blood pH or hematocrit. Cortisol levels were significantly higher in fish transported in 30 µL/L of EO of L. alba compared to those of the control group. The metabolic parameters (glycogen, lactate, total amino acid, total ammonia and total protein) showed different responses after adding EO to the transport water. In conclusion, while the EO of L. alba is recommended for fish transport in the conditions tested in the present study because it was effective in reducing waterborne total ammonia levels and net ion loss, the higher hepatic oxidative stress in this species with the same EO concentrations reported by a previous study led us to conclude that the 10–20 µL/L concentration range of EO and lack of pre-sedation before transport are more effective.     

A comparison of constant acceleration swimming speeds when acceleration rates are different with critical swimming speeds in Chinese bream under two oxygen tensions

To investigate the effect of acceleration rates on the constant acceleration test speed (U _cat) and to compare U _cat with the critical swimming speed (U _crit) in Chinese bream (Parabramis pekinensis), the U _cat test at acceleration rates of 0.05, 0.1, 0.2, 0.4 and 0.8 cm s^?2 and the U _crit test in juvenile fish at 20 °C in either normoxia (>90 % saturation oxygen tension) or hypoxia (30 % saturation) were compared. The lactate concentration ([lactate]) of white muscle, liver and plasma and the glycogen concentration ([glycogen]) of white muscle and liver were also measured to identify whether tissue substrate depletion or tissue lactate accumulation correlated with exhaustion. The U _cat decreased with the acceleration rate, and there was no significant difference between U _crit and U _cat at lower acceleration rates. Hypoxia resulted in lower U _cat and U _crit, and the difference increased with decreased acceleration rates of the U _cat test, possibly due to the increased contribution of aerobic components in U _crit or U _cat at low acceleration rates. Hypoxia elicited a significant decrease in muscle [glycogen] and an increase in muscle and liver [lactate] in resting fish. All post-exercise fish had similar muscle [lactate], suggesting that tissue lactate accumulation may correlate with exercise exhaustion. Unlike hypoxia, exercise induced an increase in muscle [lactate] and a significant increase in plasma [lactate], which were worthy of further investigation. The similar swimming speed and biochemical indicators after exercise in the U _crit and U _cat groups at low acceleration rates suggested that U _cat can be an alternative for the more frequently adopted protocols in U _crit in Chinese bream and possibly in other cyprinid fish species.     

Growing depth limit of <Emphasis Type="Italic">Zostera caulescens</Emphasis> in coastal waters along the Japan Sea coast of Honshu,Japan

There is a possibility that deep coastal marine macrophytes will be less critically affected by thermal stress associated with climate change and remain as refugia. Thus, information on them is expected to contribute to conservation of biodiversity in coastal areas affected by climate change. To document the deep-growing Zostera caulescens in relation to the light environment, field surveys were conducted in coastal waters of the central area along the Japan Sea coast of Honshu, Japan, and then the relationship between the light environment and seagrass depth limit was examined. In the coastal waters of Sado Island and Noto Peninsula, the presence of deep coastal communities of Z. caulescens was confirmed in the depth range of 20–25 m. Biomass and shoot density for Z. caulescens, ranging 34.1–51.3 g DW m^?2 and 83–112 shoots m^?2, were recorded at a depth of 20 m in Ryotsu Bay, Sado Island. For the genus Zostera including the deep-growing Z. caulescens, the relation between the extinction coefficient and seagrass depth limit was described by the fitted regression equation. The minimum light requirement of the deep-growing Z. caulescens was markedly lower than that of the shallow-growing Zostera marina.     

Effect of the early temperature on the growth of larvae and postlarvae turbot, <Emphasis Type="Italic">Scophthalmus maximus</Emphasis> L.: muscle structural and ultrastructural study

Turbot specimens were kept at three temperatures (T _s ): warm (W) (21–22 °C), ambient (A) (17–18 °C) and cold (C) (13–14 °C) during the larval and early postlarval stages. At 90 days posthatching (dph), all of them were transferred to ambient T until 190 dph. At 2–3 dph, the specimens showed a monolayer of red muscle and immature white fibres; external or dermomyotome cells (presumptive myogenic cells) were observed on the surface of the red muscle. In the following stages, many myogenic cells and presumptive myogenic precursors were observed within the myotome, presumably derived of the dermomyotome. When comparing the growth at the same age (2, 10, 25, 37 dph), the body length and the muscle growth were positively influenced by the warm T, being the hyperplasia the muscle parameter more significantly influenced. The development rate was also positively correlated with the high T: the beginning of the metamorphosis took place at 15, 23 and 25 dph at W, A and C temperatures, respectively, with the highest body length values at ambient temperature. The metamorphosis finished at 25, 30 and 37 dph at W, A and C temperatures, respectively, with the highest body length values at warm temperature. However, the muscle cellularity was similar in all the groups at the end of the metamorphosis. At 90 and 190 dph, the largest body length was observed at W temperature. However, the muscle cellularity was similar between A and W; the number of fibres was similar in all the groups at 190 dph, which shows the beginning of a compensatory muscle growth in A and C, mainly in A.     

In vitro monoamine oxidase A and B inhibitory activity and molecular docking simulations of fucoxanthin

Fucoxanthin is the primary carotenoid found in edible seaweeds, such as Eisenia bicyclis, Undaria pinnatifida, and others. The aim of this study was to determine the effectiveness of fucoxanthin against Parkinson’s disease (PD) by inhibiting monoamine oxidase (MAO) A and B because MAO inhibitors are used in early management of PD. A sensitive enzyme-based chemiluminescent assay and kinetics methods were used to investigate the enzyme inhibitory activities and mode of inhibition. A molecular docking simulation was performed to clarify the binding characteristics of fucoxanthin to hMAO-A (2z5x) and hMAO-B (2v5z). Our results suggest that fucoxanthin shows significant inhibitory activity against hMAO-A and hMAO-B with IC₅₀ values of 197.41 ± 2.20 and 211.12 ± 1.17 μM, respectively. Selegiline was used as the positive control (IC₅₀: 10.54 ± 1.25 and 0.128 ± 0.01 μM for hMAO-A and hMAO-B, respectively). The enzyme-based kinetics results demonstrated that fucoxanthin inhibited both hMAOs in a reversible competitive manner. The molecular docking simulation predicted that fucoxanthin exhibits higher binding affinity towards hMAO-A and hMAO-B through hydrogen bonding and hydrophobic interactions. These findings suggest that fucoxanthin is a reversible competitive hMAO inhibitor that could be used to manage PD.     

Quality changes of shrimp cracker covered with fish gelatin film without and with palm oil incorporated during storage

Impact of fish gelatin film incorporated without and with palm oil on the quality changes of fried shrimp cracker stored for 15 days at room temperature was investigated, in comparison with nylon/linear low-density polyethylene (nylon/LLDPE) film. The moisture content and water activity of shrimp cracker packaged with all films increased during storage (p < 0.05). The lowest moisture content and water activity were found in the sample packaged with nylon/LLDPE film throughout the storage (p < 0.05). Sample packaged with fish gelatin films incorporated with palm oil generally had lower moisture content than those without oil added during the first 12 days of storage (p < 0.05). During 15 days of storage, shrimp cracker packaged with nylon/LLDPE film generally had the lower PV and TBARS value as well as volatile compounds, except for n-nonanal, than those stored in fish gelatin films, regardless of oil incorporation. The decrease in crispiness and increase in toughness occurred in all samples during the 15 days of storage. Nevertheless, the lower changes were observed in the sample packaged with nylon/LLDPE film. Overall, gelatin film showed excellent oxygen barrier property, which was associated with the retardation of lipid oxidation. The incorporation of oil into gelatin film could lower WVP, but negatively increased oxygen permeability of the resulting film. Thus, the improvement of gelatin film is still required.     

Effects of temperature and fatigue on the metabolism and swimming capacity of juvenile Chinese sturgeon (<Emphasis Type="Italic">Acipenser sinensis</Emphasis>)

Chinese sturgeon (Acipenser sinensis) is a critically endangered species. A flume-type respirometer, with video, was used to conduct two consecutive stepped velocity tests at 10, 15, 20, and 25 °C. Extent of recovery was measured after the 60-min recovery period between trials, and the recovery ratio for critical swimming speed (U _crit) averaged 91.88% across temperatures. Temperature (T) effects were determined by comparing U _crit, oxygen consumption rate (MO ₂), and tail beat frequency (TBF) for each temperature. Results from the two trials were compared to determine the effect of exercise. The U _crit occurring at 15 °C in both trials was significantly higher than that at 10 and 25 °C (p < 0.05). The U _crit was plotted as a function of T and curve-fitting allowed calculation of the optimal swimming temperature 3.28 BL/s at 15.96 °C (trial 1) and 2.98 BL/s at 15.85 °C (trial 2). In trial 1, MO ₂ increased rapidly with U, but then declined sharply as swimming speed approached U _crit. In trial 2, MO ₂ increased more slowly, but continuously, to U _crit. TBF was directly proportional to U and the slope (dTBF/dU) for trial 2 was significantly lower than that for trial 1. The inverse slope (tail beats per body length, TB/BL) is a measure of swimming efficiency and the significant difference in slopes implies that the exercise training provided by trial 1 led to a significant increase in swimming efficiency in trial 2.     

The effects of starvation and re-feeding on growth and swimming performance of juvenile black carp (<Emphasis Type="Italic">Mylopharyngodon piceus</Emphasis>)

We investigated the effects of starvation and re-feeding on growth and swimming performance and their relationship in juvenile black carp (Mylopharyngodon piceus). We measured the specific growth rate (SGR), resting metabolic rate (RMR) and constant acceleration test speed (U _CAT, the maximum swimming speed at exhaustion by constant acceleration test with 0.1667 cm s^?2 rate) in a treatment group (21 days of starvation then 21 days of re-feeding) and control group (routine feeding) (n = 20). Starvation resulted in a 17 % decrease in body mass of black carp (P < 0.05). After 21 days of re-feeding, body mass was greater than that of pre-starvation but still less than that of the control group at 42 days. During the re-feeding phase, the SGR of the treatment group was higher than that of the control group (P < 0.05). Starvation resulted in a significant decrease in the RMR and U _CAT. After 21 days of re-feeding, both the RMR and U _CAT recovered to the pre-starvation levels. In the control group, individual juvenile black carp displayed strong repeatability of the RMR and U _CAT across the measurement periods (P ≤ 0.002). In the treatment group, RMR showed significant repeatability between pre-starvation and re-feeding (P = 0.007), but not between pre-starvation and starvation or between starvation and re-feeding. U _CAT showed significant repeatability between pre-starvation and starvation (P = 0.006) and between pre-starvation and re-feeding (P = 0.001), but not between starvation and re-feeding. No correlation or only a weak correlation was found between any two variables of RMR, U _CAT and SGR, whereas the increment of the U _CAT (ΔU _CAT) was negatively correlated with that of SGR during the starvation phase (r = ?0.581, n = 20, P = 0.007) and re-feeding phase (r = ?0.568, n = 20, P = 0.009). This suggested that within individual black carp, there is a trade-off between growth and maintenance (or development) of swimming performance under food-limited conditions.     

Astaxanthin degradation and lipid oxidation of Pacific white shrimp oil: kinetics study and stability as affected by storage conditions

The kinetics of astaxanthin degradation and lipid oxidation in shrimp oil from hepatopancreas of Pacific white shrimp (Litopenaeus vannamei) as affected by storage temperature were studied. When shrimp oil was incubated at different temperatures (4, 30, 45 and 60 °C) for 16 h, the rate constants (k) of astaxanthin degradation and lipid oxidation in shrimp oil increased with increasing temperatures (p < 0.05). Thus, astaxanthin degradation and lipid oxidation in shrimp oil were augmented at high temperature. When shrimp oils with different storage conditions (illumination, oxygen availability and temperature) were stored for up to 40 days, astaxanthin contents in all samples decreased throughout storage (p < 0.05). All factors were able to enhance astaxanthin degradation during 40 days of storage. With increasing storage time, the progressive formation of primary and secondary oxidation products were found in all samples as evidenced by the increases in both peroxide values (PV) and thiobarbituric acid reactive substances (TBARS) (p < 0.05). Light, air and temperatures therefore had the marked effect on astaxanthin degradation and lipid oxidation in shrimp oils during the extended storage.     

Effect of biofilm on water quality and growth of <Emphasis Type="Italic">Etroplus suratensis</Emphasis> (Bloch, 1790)

An experiment was conducted to compare the efficiency of biofilm production in natural and artificial substrates and to study their effect on water quality and growth of Etroplus suratensis. Four different substrates were used for biofilm formation: paddy straw (T1), sugarcane bagasse (T2), polyvinylchloride (PVC) pipe (T3) and plastic sheet (T4). The experiment was carried out in mud-bottomed fibre-reinforced plastic tanks (300 L) in triplicates. About 3000 cm² surface area (600 g) of each substrate was suspended in water supplemented with fertilizers. Only cow dung and urea were applied in control tanks. The tanks were stocked with 25 fishes with average weight of 9.1 ± 0.22 g. The overall mean value of heterotrophic bacteria in substrate was found higher in straw followed by bagasse, plastic and PVC. The dominant genera of bacteria in the substrate were Bacillus, Pseudomonas and Micrococcus in that order of preponderance. The mean phytoplankton and zooplankton density on the substrates were higher in bagasse followed by straw, plastic and PVC. The biofilm developed on the substrate significantly reduced the ammonia nitrogen and nitrite-nitrogen content of water. The growth of fishes was significantly (P < 0.05) higher in substrate-based treatments than that in the control with better results in bagasse followed by straw, plastic and PVC. The conclusions of the present study are that biofilm produced on natural substrates, especially on bagasse, enhanced growth of E. suratensis and reduced the necessity of water exchange during the culture, which certainly decreases the cost of Etroplus production.     

Microalga <Emphasis Type="Italic">Isochrysis galbana</Emphasis> in feed for <Emphasis Type="Italic">Trachinotus ovatus</Emphasis>: effect on growth performance and fatty acid composition of fish fillet and liver

Seeking alternatives to the depleting fish oil are crucial for marine fish aquaculture, which is currently dependent on fish oil as the primary source of n-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFAs). Five isonitrogenous (46% crude protein) and isolipidic (16% crude lipid) feed diets (FO, ISO2.9, ISO4.8, ISO6.7, ISO8.6) were formulated by partially replacing fish oil with microalgae Isochrysis galbana. These diets were fed to triplicate tanks of Trachinotus ovatus (mean initial weight 1.92 g) for 80 days. This work demonstrates that a moderate inclusion (around 4.5–5.0 wt%, equivalent to the replacement of 24–26 wt% fish oil) of I. galbana biomass in fish diet improves fish growth performance, lipid deposition and enhances total n-3 fatty acids, DHA, and EPA contents in neutral and polar lipids (PLs) of fish muscle and liver of T. ovatus. The results disclosed in this study suggest that I. galbana microalgae represents a potential high-quality substitute for fish-based ingredients in aquaculture feeds, which can be a promising sustainable solution to resolve the depleting fish oil resource in a cost-effective manner.     

Influence of the dietary inclusion of <Emphasis Type="Italic">Gracilaria cornea</Emphasis> and <Emphasis Type="Italic">Ulva rigida</Emphasis> on the biodiversity of the intestinal microbiota of <Emphasis Type="Italic">Sparus aurata</Emphasis> juveniles

Partial substitutions of fish meal by 5, 15, or 25 % of Gracilaria cornea or Ulva rigida in experimental diets were evaluated to study their effects on biodiversity of intestinal microbiota composition in gilthead seabream (Sparus aurata) juveniles. The diets were offered to duplicate groups of 15 juvenile fish (14.0 ± 0.5 g) for 70 days, and at the end of the experiment the intestinal microbiota from four specimens of each treatment was analysed by denaturing gel gradient electrophoresis. Results showed that the substitution of fish meal by algae meal induced important modifications in the intestinal microbiota community, as a big reduction of the biodiversity when the highest percentage (25 %) of U. rigida was included. On the contrary, an increase on the number of species was detected when a 15 % of algae was included. Various Lactobacillus delbrueckii subspecies were selectively stimulated when G. cornea was included in the feed, and other bacterial species, such as those included in the Vibrio genus, were reduced.     

Comparative nutritional value of <Emphasis Type="Italic">Jatropha curcas</Emphasis> protein isolate and soy protein isolate in common carp

Jatropha seed cake (JSC) is an excellent source of protein but does contain some antinutritional factors (ANF) that can act as toxins and thus negatively affect the growth and health status of fish. While this can limit the use of JSC, detoxified Jatropha protein isolate (DJPI) may be a better option. An 8-week study was performed to evaluate dietary DJPI to common carp Cyprinus carpio. Five iso-nitrogenous diets (crude protein of 38%) were formulated that consisted of a C _ontrol (fish meal (FM) based protein), J ₅₀ or J ₇₅ (50 and 75% of FM protein replaced by DJPI), and S ₅₀ or S ₇₅ (50 and 75% of FM protein replaced by soy protein isolate, SPI) and fed to triplicate groups of 75 carp fingerlings (75; av. wt. ± SD; 11.4 ± 0.25 g). The growth, feeding efficiencies, digestibility, plasma biochemistry, and intestinal enzymes were measured. Results showed that growth performance of fish fed the S ₇₅- or DJPI-based diets were not significantly different from those fed the C _ontrol diet, while carp fed the S ₅₀ had significantly better growth than the J ₇₅ diet. Fish fed the J ₇₅ diet had significantly lower protein and lipid digestibility as well as significantly lower intestinal amylase and protease activities than all other groups. However, all plant protein-based diets led to significantly higher crude protein, crude lipid, and gross energy in the body of common carp compared to the control treatment. Plasma cholesterol and creatinine significantly decreased in the plant protein fed groups, although plasma triglyceride as well as the red blood cells count, hematocrit, albumin, globulin, total plasma protein, and lysozyme activity were higher in plant protein fed groups compared to FM fed group. White blood cells, hemoglobulin concentration, alkaline phosphatase and alanine transaminase activities, and glucose level in blood did not differ significantly among treatments. The results suggest that the DJPI is non-toxic to carp and can be used to replace FM in the diets of common carp up to 75%, but further research to potentially reduce some inherent ANF within this protein source, such as non-starch polysaccharides, may improve nutrient utilization.