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.     

Microalgal paste production of the diatom <Emphasis Type="Italic">Chaetoceros calcitrans</Emphasis> using electrolytic flocculation method at optimum culture conditions

The optimum culture conditions of the local strain Chaetoceros calcitrans were determined to improve biomass and reduce cost of production. Under outdoor culture conditions, higher cell density was attained when the cultures were enriched with Tungkang Marine Research Laboratory (TMRL) medium composed of cheap technical grade reagents and cultured at 25 g L^?1 salinity. The cultures were lighted with two 40 W cool-white GE fluorescent tubes (24–35 μmol photon m^?2 s^?1). Using semi-continuous culture system under established optimum culture conditions, C. calcitrans can be re-cultured thrice and concentrated at each culture cycle using electrolytic flocculation method to produce 4.6 kg m^?3 of diatom paste. The viability of concentrated C. calcitrans after 3 months of storage was comparable to live diatom cells. Simple preservation technique by low-temperature storage is convenient for storing algal concentrates for use as starter cultures and for feeding invertebrates. The paste costs USD 8.24 kg^?1 inclusive of the assets and flocculation materials for culturing and harvesting the diatom, respectively. This study established the suitable conditions for mass culture of C. calcitrans and produced concentrated diatoms in paste form that is readily available for aquaculture hatcheries at a lower cost.     

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.     

Effect of irradiance on bioremediation capacity of the red algae Chondrus crispus and Palmaria palmata

To investigate the illuminance requirements for the culture of red algae in finfish effluent, both Chondrus crispus and Palmaria palmata were assessed simultaneously at three irradiances. Apical tips, 3.5 cm length, were stocked at 2 g L^?1 in 1-L glass flasks with enriched seawater in two 24-day experiments, at 10 and 14 °C, respectively. Palmaria grew up to 70 % better and up took more nutrients than Chondrus under five of the six experimental conditions, except at 14 °C and low irradiance (daily photon dose of 3.4 mol photon m^?2 day^?1). Specific growth rate of both species at high irradiance, 13.1 and 17.2 mol photon m^?2 day^?1, at each trial, 10 and 14 °C, respectively, was 40 and 20 % better than at medium irradiance, but nutrient uptake was similar at both medium and high irradiance. Chondrus growth rate was higher during Experiment 2 (14 °C and 17.2 mol photon m^?2 day^?1) than during Experiment 1 (10 °C and 13.1 mol photon m^?2 day^?1), while Palmaria growth rate was similar in both trials, but always significantly higher than Chondrus. In the range 10–14 °C, Palmaria is the best candidate for integration with Atlantic halibut juveniles (Hippoglossus hippoglossus).     

The effect of irradiance and temperature on the photosynthesis of two agarophytes Gelidium elegans and Pterocladiella tenuis (Gelidiales) from Kagoshima,Japan

The effect of irradiance and temperature on the photosynthesis of two Japanese agarophytes, Gelidium elegans and Pterocladiella tenuis (Gelidiales), was determined using dissolved oxygen sensors and pulse amplitude modulated (PAM) fluorometry. Gross photosynthesis and dark respiration rates were determined over a range of temperatures (8–36 °C). The highest gross photosynthetic rates were 40.3 and 37.0 mg O₂ g _ww ^?1  min^?1 and occurred at 24.3 and 25.5 °C [95 % Bayesian credible interval (BCI) 20.7–28.0 and 23.4–28.3 °C], respectively. The dark respiration rate in G. elegans and P. tenuis increased with increasing temperature at a rate of 0.10 and 0.31 mg O₂ g _ww ^?1  min^?1 °C^?1 , respectively. Modeling the net photosynthesis–irradiance (P–E) responses of G. elegans and P. tenuis at 20 °C revealed that the net photosynthetic rates quickly increased at irradiance levels below the estimated saturation irradiance of 88 and 83 µmol photons m^?2 s^?1, with a compensation irradiance of 14 and 19 µmol photons m^?2 s^?1, respectively. The highest value of the maximum effective quantum yield (Φ _PSII) occurred at 20.1 °C (BCI 18.9–21.5 °C) and 21.3 °C (BCI 20.2–22.5 °C) for G. elegans and P. tenuis and was 0.49 and 0.45, respectively. These optimal temperatures of Φ _PSII were relatively lower than those determined by the photosynthesis–temperature model of oxygen evolution. The temperature response of these species indicates that they are probably well adapted to the current range of seawater temperatures in the study site but that they are near the boundary of their tolerable limits.     

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.     

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.     

Biofloc management with different flow rates for solids removal in the <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> BFT culture system

Different water flows for solids removal in the Litopenaeus vannamei BFT system were evaluated. One control (no solids removal) and two treatments using different water flows, high (3945 L h^?1—HF) and low (1750 L h^?1—LF), were used with no water replenishment after each process, and the total dry weight of the solids was measured. L. vannamei (0.18 ± 0.06 g; 350 individuals m^?2) were stocked in 35-m^?3 tanks. For 17 weeks, the physical and chemical parameters were maintained within the recommended. To keep the total suspended solids concentrations at approximately 500–600 mg L^?1, clarifying was performed. The average water volume flowed by clarifiers was significantly different (p < 0.05) between HF (205 ± 34 m³) and LF (114 ± 24 m³). There was a significant decrease (p < 0.05) in the final tank volume in HF (28.09 ± 0.92 m³) and LF (28.62 ± 1.38 m³) due to the clarifying. Before clarifying (initial sample) and at the end of experiment (final sample) were not significantly different (p > 0.05) for crude protein, moisture or ash. The crude lipid of the LF in the final period was significantly lower (p < 0.05) compared to others in both periods. The survival, productivity and food conversion ratio were significantly better (p < 0.05) in the HF and LF treatments compared to those of the control. The best shrimp performance was obtained with solids removal. The lower flow in the clarifier facilitated particle settling, allowing adjustment of the flow.     

Demographic and competition studies on <Emphasis Type="Italic">Brachionus ibericus</Emphasis> and <Emphasis Type="Italic">Proales similis</Emphasis> in relation to salinity and algal (<Emphasis Type="Italic">Nannochloropsis oculata</Emphasis>) density

We isolated the rotifers Brachionus ibericus and Proales similis from the sediment of shrimp tanks and studied their individual demographic characters and competition between them at two food levels (0.25?×?10⁶, 1.00?×?10⁶ cells ml^?1 of Nannochloropsis oculata at 25 °C) and salinities ranging from 10 to 30‰. Our hypothesis was that growth rates would be higher with increasing food levels and salinities. Observations were taken twice a day for life table studies and daily once for population growth experiments. Using survivorship and fecundity data, we derived various life history variables. Although the average life span (7.6?±?0.4 days) and gross reproductive rate (33.8?±?2.9 neonate female^?1 day^?1) of B. ibericus were higher than those of P. similis (average life span 5.4?±?0.6 days and gross reproductive rate 13.0?±?0.6 neonate female^?1 day^?1), the population growth experiments showed that P. similis had higher r values (0.32?±?0.005 day^?1) than B. ibericus (0.23?±?0.002 day^?1) at 1.0?×?10⁶ cells ml^?1 of N. oculata. The rotifer P. similis was more adversely affected due to the presence of B. ibericus than vice versa. The data are important for developing techniques for a large-scale culture of these rotifers as food in aquaculture.     

Alternative microalgal diets for cultivation of the fairy shrimp <Emphasis Type="Italic">Branchinella thailandensis</Emphasis> (Branchiopoda: Anostraca)

Fairy shrimp is known as a nutritional food for fish and crustaceans in aquaculture. In most hatcheries, the microalga Chlorella sp. appears to be the most common, suitable, and nutritious food to feed fairy shrimp. In this study, we attempted to determine other alternative algal diets for cultivation of fairy shrimp Branchinella thailandensis. Seven experimental diets including three treatments of dried Spirulina sp. at 0.75 (S1), 1.5 (S2), and 3.0 mg dry weight individual^?1 (S3); three treatments of Chlorococcum humicola at 5 × 10⁵ (Ch1), 1 × 10⁶ (Ch2), and 2 × 10⁶ cells mL^?1 (Ch3); and a control diet (Chlorella vulgaris at 1 × 10⁶ cells mL^?1) were fed to 5-day-old shrimp for 15 days. Evaluation of growth performance, egg production, survival percentage, and nutritional and carotenoid content of the experimental fairy shrimp revealed that Ch3 is the most suitable algal diet. Our results suggest that C. humicola is the best alternative food source for the cultivation of B. thailandensis. In addition, dried Spirulina powder is also a good choice when live algae are not available and can be used as an alternative feed in fairy shrimp cultures.     

Corn gluten meal in pacu <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> diets: effects on growth,haematology, and meat quality

Corn gluten meal (CGM) has a high protein content and absence of antinutrients. However, it has high levels of carotenoids that can cause a yellowing of fish fillets and impair further commercialisation. Graded levels of CGM were incorporated in pacu diets to replace soybean meal (SM) protein. The experimental diets were formulated to be isonitrogenous (220 g kg^?1 digestible protein) and isoenergetic (13.4 MJ kg^?1 digestible energy) with 0, 25, 50, 75, and 100 % replacement of SM protein by CGM protein over a 60-day period. One hundred fish (32.79 ± 3.43 g) were randomly distributed in 20 experimental cages (70 L) placed within five (1000 L) aquaria with continuous water renewal. The experimental design was completely randomised with five treatments and four replicates. A quadratic effect (P < 0.05) was observed for fillet and carcass yields, feed conversion ratio, weight gain, and specific growth rate, with the optimum values for replacement of SM protein by CGM protein estimated as 21.95, 29.13, 30.94, 37.71, and 38.75 %, respectively. There were no differences (P > 0.05) for blood parameters, proximal composition, water-holding capacity, pH, lipid oxidation, and a* value of fillets. The L* and b* values of fillets showed differences (P < 0.05). Replacement between 21.95 and 38.75 % of SM protein by CGM protein (5.37 and 9.48 % of inclusion in the diet) improves the growth performance and body yield of pacu juveniles without altering blood parameters or affecting the fillet quality.     

Effects of dietary <Emphasis Type="Italic">myo</Emphasis>-inositol on growth,chemical composition and plasma chemistry of Amur sturgeon <Emphasis Type="Italic">Acipenser schrenckii</Emphasis>

The present study was conducted to demonstrate the dietary myo-inositol requirement and its effects on the growth, proximate composition and blood chemistry of Amur sturgeon (Acipenser schrenckii). Triplicate groups of 30 fish (initial weight 11.90?±?0.12 g) were fed different diets containing graded levels of myo-inositol (28.75, 127.83, 343.83, 565.81, 738.15 and 936.28 mg kg^?1) until satiation for 56 days. The fish were weighed after a 24-h fast, and six fish were used for whole body composition analysis. Further, the liver and muscle were sampled from another six fish for lipid analysis. The blood and liver were sampled from the remaining six fish for haematology and fatty acid analysis. The weight gain of fish increased with myo-inositol content, from the 28.75- to 343.83-mg kg^?1 myo-inositol treatment groups, and then stabilised. The liver lipid content and hepatosomatic index decreased significantly from 21.91 to 19.14% and from 3.20 to 2.76% with increased dietary myo-inositol supplementation, respectively. The whole body lipid content generally decreased from 6.33 to 5.55%. The content of liver-saturated fatty acids decreased significantly (28.13%) in the 936.28-mg kg^?1 treatment group. The content of plasma non-esterified fatty acids increased with the increase in dietary myo-inositol supplementation from 0.77 to 1.17 mmol L^?1, whereas the content of triglycerides significantly decreased from 4.62 to 3.28 mmol L^?1. In conclusion, the optimum myo-inositol requirement was found to be 336.1 mg kg^?1, based on weight gain in a two-slope quadratic broken-line model.     

Transcripts of genes encoding reproductive neuroendocrine hormones and androgen receptor in the brain and testis of goldfish exposed to vinclozolin,flutamide, testosterone,and their combinations

Vinclozolin (VZ) is a pesticide that acts as an anti-androgen to impair reproduction in mammals. However, VZ-induced disruption of reproduction is largely unknown in fish. In the present study, we have established a combination exposure in which adult goldfish were exposed to VZ (30 and 100 μg/L), anti-androgen flutamide (Flu, 300 μg/L), and androgen testosterone (T, 1 μg/L) to better understand effects of VZ on reproductive endocrine system. mRNA levels of kisspeptin (kiss-1 and kiss-2) and its receptor (gpr54), salmon gonadotropin-releasing hormone (gnrh3) and androgen receptor (ar) in the mid-brain, and luteinizing hormone receptor (lhr) in the testis were analyzed and compared with those of control following 10 days of exposure. kiss-1 mRNA level was increased in goldfish exposed to 100 µg/L VZ and to Flu, while kiss-2 mRNA level was increased following exposure to Flu and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. gpr54 mRNA level was increased in goldfish exposed to Flu and to combination of 30 µg/L VZ with Flu and 100 µg/L VZ with T. gnrh3 mRNA level was increased in goldfish exposed to 100 µg/L VZ, to Flu, and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. The mid-brain ar mRNA level was increased in goldfish exposed to Flu and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. Testicular lhr mRNA level was increased in goldfish exposed to Flu and to combination of 30 µg/L VZ with Flu. These results suggest that VZ and Flu are capable of interfering with kisspeptin and GnRH systems to alter pituitary and testicular horonal functions in adult goldfish and the brain ar mediates VZ-induced disruption of androgen production.     

In vitro and in vivo evaluation of probiotic properties of <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> in Kenyi cichlid, <Emphasis Type="Italic">Maylandia lombardoi</Emphasis>

In this study, Enterobacter cloacae bacterium was isolated from curd and its antibacterial potential against the pathogen Plesiomonas shigelloides was evaluated using the freshwater ornamental fish Kenyi cichlid (Maylandia lombardoi). Among the bacterial isolates, E. cloacae exhibited tolerance to acidic pH 2 and demonstrated the highest antibacterial activity against P. shigelloides in various in vitro assays. Dietary supplementation of E. cloacae with prebiotic supplement 2% mannan oligosaccharide improved the growth performance and reduced the toxic metabolites such as nitrite in culture tank water of Kenyi cichlid. In vivo application of E. cloacae with mannan oligosaccharide significantly elevated (p?<?0.05) white blood cell counts (88.47?±?2.15 10³ mm^?3) and respiratory burst activity (0.243?±?0.007) of Kenyi cichlid when challenged with P. shigelloides. Intestinal morphology of Kenyi cichlid fish treated with probiotic and prebiotic combinations showed improved intestinal architectures. The present findings confirm that the isolated bacterium E. cloacae is a potential probiotic and can be used effectively to prevent infection of P. shigelloides in freshwater ornamental fish culture.     

Effects of nucleotides on growth performance,immune response,disease resistance and intestinal morphology in shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> fed with a low fish meal diet

Nucleotides (NT) are widely used as an immunostimulant in aquaculture. A 10-week feeding trial was conducted to evaluate the effects of dietary NT on the growth performance, immune response, disease resistance and intestinal morphology of the Pacific white shrimp Litopenaeus vannamei (initial mean body weight: 0.39 ± 0.00 g). Five isolipidic (about 7 % crude lipid) and isonitrogenous (about 39 % crude protein) practical diets were supplemented with graded levels of NT (0, 60, 90, 120 and 1200 mg kg^?1), respectively. These diets were named as N1, N2, N3, N4 and N5. Each diet was randomly fed to six tanks of shrimps, and each tank contained 40 shrimps. The shrimps were fed four times daily (07:00, 11:00, 16:00 and 21:00 h). The results showed that there were no significant differences in survival, final weight, specific growth rate, feed intake, feed conversion ratio and whole-body compositions among all the treatments (P > 0.05). Shrimps fed the control diet had the significantly lowest activities of superoxide dismutase, total nitric oxide synthase and lysozyme (P < 0.05). However, those parameters increased when dietary NT increased from 60 to 120 mg kg^?1 and decreased as inclusion level increased to 1200 mg kg^?1 (P < 0.05). The cumulative mortality of the shrimps challenged with Vibrio parahaemolyticus was significantly higher in the treatments without dietary NT supplementation than those in treatments with dietary NT (≥90 mg kg^?1) (P < 0.05). The jejunum wall thickness in shrimps fed the control diet was significantly lower than that in the treatments of N2, N3 and N4 (P < 0.05). For the villus height, the highest value (49.29 µm) was found in N3. In summary, the present study showed that 90 mg kg^?1 of dietary NT is the optimum dietary level for good gut health, immune response and disease resistance of Pacific white shrimp fed a diet with 18 % fish meal.     

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.     

Effect of stocking density on growth performance,serum biochemical parameters,and muscle texture properties of genetically improved farm tilapia, <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

The objective of this study was to assess the effects of stocking density on growth performance, serum biochemical parameters, and muscle texture properties of genetically improved farmed tilapia (Oreochromis niloticus, GIFT). Juvenile GIFT with an average initial weight of 12.54?±?0.45 g (mean?±?SD) were randomly stocked in 16 tanks (80 L) in a recirculation aquaculture system at four densities of 10 (D1), 20 (D2), 30 (D3), and 40 (D4) fish per tank for 56 days, with quadruplicate for each density. There were no significant differences in water temperature among the four treatments (P?>?0.05). D4 had the significantly lowest dissolved oxygen content (5.52 vs 5.69–6.09 mg L^?1) (P?>?0.05) and pH (6.63 vs 6.87–7.20) (P?<?0.05). NO₂-N and NH₄-N concentrations significantly increased with increasing stocking density (P?<?0.05). Weight gain (WG) and specific growth rates (SGR) decreased with increasing stocking density. The lowest WG (617.20 vs 660.45–747.06%), SGR (3.52 vs 3.62–3.81% day^?1), and highest feed conversion ratio (1.68 vs 1.53–1.58) were observed in D4. Fish at D4 had significantly lower condition factor (3.11 vs 3.29–3.37%) and survival rate (91.25 vs 97.50%) than those from D1 and D2 (P?<?0.05). With increasing stocking density, serum total cholesterol, triglyceride, and total protein concentrations decreased (P <?0.05) and aspartate aminotransferase and alanine aminotransferase activities increased (P <?0.05). D4 fish had higher moisture content (78.80 vs 76.97%) and lower crude protein content (18.14 vs 19.39%) in muscle than D1 fish (P?<?0.05). Compared to D1 and D2, D3 and D4 had lower muscle hardness (1271.54–1294.07 vs 1465.12–1485.65 g), springiness (0.62–0.65 vs 0.70–0.72), gumminess (857.33–885.32 vs 1058.82–1079.28 g), and chewiness (533.04–577.09 vs 757.53–775.69 g) (P <?0.05). High stocking density resulted in growth inhibition, declines in flesh quality, and disturbance to several serum biochemical parameters.     

The changes in cortisol and expression of immune genes of GIFT tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> (L.) at different rearing densities under <Emphasis Type="Italic">Streptococcus iniae</Emphasis> infection

The present study was conducted to investigate the effects of stocking density on serum cortisol (COR) levels and expression of immune genes in the head kidney of juvenile GIFT tilapia (Oreochromis niloticus) after Streptococcus iniae (S. iniae) infection. Juveniles (2700) were distributed into 30 tanks at five stocking densities (150, 300, 450, 600 and 750 g/m³), and each treatment had six replicates. After a 45-day feeding trial, a S. iniae challenge study was conducted for 96 h. Differences in mortality, serum COR and gene expression of C-type lysozyme, heat-shock protein 70 (HSP70), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) were analyzed. The levels of serum COR at all infected treatments were significantly higher compared with the levels of pre-infection (P < 0.05). At 96 h post-infection, serum COR levels of high stocking densities (600 and 750 g/m³) were significantly higher than those of low-density groups (P < 0.05); the mRNA levels of C-type lysozyme, HSP70, IL-1β, TNF-α and IFN-γ were significantly lower in tilapia reared at 600 g/m³ group than in those reared at 300 g/m³ group (P < 0.05). Fish reared in high-density treatments (600 and 750 g/m³) exhibited significantly higher post-challenge mortality (43.3 and 40.0 %) at 96 h following S. iniae infection, and it was significantly higher than other groups (P < 0.05). The data suggest that when living in high stocking density, GIFT tilapia showed decrease in immune capability, together with increased serum COR and susceptibility to S. iniae.     

Immunomodulatory effects of <Emphasis Type="Italic">Rhodomyrtus tomentosa</Emphasis> leaf extract and its derivative compound,rhodomyrtone, on head kidney macrophages of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

Rhodomyrtus tomentosa is a medicinal plant that shows biological effects including immunomodulatory activity on human and other mammals but not in fish. In this study, we evaluated the in vitro immunomodulatory effects of R. tomentosa leaf extract and its active compound, rhodomyrtone, on the immune responses, using rainbow trout (Oncorhynchus mykiss) head kidney (HK) macrophages as a model. The tested immune functions included the expression of genes involved in innate immune and inflammatory responses and the production of reactive oxygen species (ROS). Gene expression was evaluated after exposure to 10 μg mL^?1 of R. tomentosa and 1 μg mL^?1 of rhodomyrtone for 4 and 24 h. R. tomentosa and rhodomyrtone induced changes in the expression of pro-inflammatory cytokines (il1β, il8, and tnfα), anti-inflammatory cytokines (il10 and tgfβ), inducible enzymes (inos, cox2, and arginase), and an antioxidant enzyme (gpx1). Co-exposure of R. tomentosa with LPS resulted in a prominent reduction in the expression of genes related to an inflammatory process (il1β, il8, tnfα, inos, saa, hepcidin, and gpx1), suggesting anti-inflammatory effects. Similarly, co-exposure of rhodomyrtone with LPS led to a downregulation of inflammation-related genes (il1β, inos, saa, and hepcidin). In addition, exposure to both natural plant products caused a reduction in cellular ROS levels by HK macrophages. The present results indicate that R. tomentosa and rhodomyrtone exerted immunostimulatory and anti-inflammatory effects on fish macrophages, thus opening up the possibility of using these natural products to further develop immunostimulants for health management in aquaculture.     

Effect of dietary chitosan on growth performance,hematological parameters,intestinal histology and stress resistance of Caspian kutum (<Emphasis Type="Italic">Rutilus frisii kutum</Emphasis> Kamenskii, 1901) fingerlings

This study was conducted to investigate the effects of dietary chitosan on growth performance, hematological parameters, intestinal histology, stress resistance and body composition in the Caspian kutum (Rutilus frisii kutum, Kamenskii, 1901) fingerlings. Fish (1.7 ± 0.15 g) were fed diets containing chitosan at different levels (0, 0.25, 0.5, 1 and 2 g kg^?1 diet) for a period of 60 days. Results showed that the feed conversion ratio significantly decreased in fish fed diet containing 1 g kg^?1 of chitosan compared to the other groups (P < 0.05), but there were no significant differences between treatments in terms of specific growth rate and condition factor (P > 0.05). Leukocyte increased in fish fed diet containing 2 g kg^?1 of chitosan compared to the other groups (P < 0.05). Lymphocytes, eosinophils and neutrophils did not significantly change among dietary treatments (P > 0.05). Also, the chitosan supplementation did not affect the whole-fish body composition (P > 0.05). Light microscopy demonstrated that the intestinal villus length increased in fish fed diet containing 1 g kg^?1 of chitosan compared to control group (P < 0.05). While 11 and 13 ‰ salinity and 30 °C thermal stress had no effect, 1 g kg^?1 of chitosan (P < 0.05) showed highest survival rate (70 %) in 34 °C thermal stress. The results showed that chitosan in the diet of the Caspian kutum fingerlings could improve feed conversion ratio, the nonspecific defense mechanisms and resistance to some of the environmental stresses.