Onion Powder in the Diet of the Olive Flounder,Paralichthys olivaceus: Effects on the Growth,Body Composition,and Lysozyme Activity

This study tested the onion powder (OP) supplementation in the diet of the olive flounder on the growth, body composition, and lysozyme activity. Thirty‐five fish averaging 5.1 g were randomly stocked into 18 individual 180‐L flow‐through tanks. A commercially available OP was used as a dietary additive. Six experimental diets were prepared to contain OP at the concentrations of 0, 0.5, 1, 2, 3, and 5% for diets OP‐0, OP‐0.5, OP‐1, OP‐2, OP‐3, and OP‐5, respectively. After the 8‐wk feeding trial, 20 fish from each tank were infected with Edwardsiella tarda and mortality was monitored for the following 96 h. No distinctive improvement in survival, weight gain, or feed efficiency of fish was observed at the end of the 8‐wk feeding trial. Lysozyme activity in fish fed the OP‐0.5 diet was higher than that of fish fed the OP‐0, OP‐1, OP‐2, OP‐3, and OP‐5 diets. The cumulative mortality of fish fed the OP‐0.5, OP‐1, OP‐2, OP‐3, and OP‐5 diets was lower than that of fish fed the OP‐0 diet at 72 h after E. tarda infection. Dietary inclusion of 0.5% OP was effective at improving lysozyme activity of fish, and OP seemed to be an effective immunostimulant to lower mortality upon E. tarda infection.     

Effects of propolis enriched diet on growth performance and plasma biochemical parameters of juvenile European sea bass (Dicentrarchus labrax L.) under acute low‐temperature stress

To study the effect of propolis in crude form (CPP), prepared without any chemical refinement (CPP) on Dicentrarchus labrax under low‐temperature stress, sea bass juveniles were randomly divided into three groups: a control group fed with basal diet and two treatment groups fed with basal diet supplemented with 1.25 and 2.5 g kg⁻¹ of propolis. At the end of a 10‐week feeding trial, sea bass were exposed to low‐temperature stress at 12 °C for 24 h. The growth performance, RNA/DNA ratio and changes in serum biochemical parameters were investigated. Dietary intake of propolis stimulated the specific growth rate (SGR), feed conversion efficiency (FCE), RNA/DNA ratio and alkaline phosphatase (ALP) enzyme activity, while decreasing plasma triglycerides and aspartate aminotransferase (AST) activity. Supplement of 2.5 g kg⁻¹ CPP in diet significantly increased the mean SGR and FCE up to 9% and 13.4%, respectively, in comparison with the control group. Low‐temperature stress elevated serum triglycerides, glucose and cortisol levels in all groups; however, glucose and cortisol reached significantly lower end values in group receiving highest amount of propolis in diet. This study suggests that ingestion of basal diet supplemented with 2.5 g kg⁻¹ of propolis could prevent adverse effects of low‐temperature stress and promote the growth of sea bass.     

Comparative Efficacy of Anesthetics for the Freshwater Prawn Macrobrachiurn rosenbergii

Abstract— The freshwater prawn Macrobrachiurn rosenbergii is a commercially important culture species in the South Central United States. Two major constraints in the commercial culture of the freshwater prawn in the U.S. are poor survival during live transportation of seed‐stock to growout ponds, and live transportation of pond harvested prawn to distant live markets due to the territorial and cannibalistic nature of prawn. The use of anesthetics could possibly improve transport survival; however, to date anesthetic agents have not been evaluated for use with prawn. Two trials were conducted with juvenile freshwater prawn to compare the efficacy of anesthetics commonly used on fish. The first trial was designed to identify the most promising candidates. In Study 1, tricaine methanesulfonate (MS‐222), 2‐phenoxyethanol, quinaldine sulfate (quinaldine), clove oil, and Aqui‐S^TM were evaluated at 25 and 100 mg/L for 1 h in three replicate 10‐L glass containers, containing five juvenile prawn each. Relative sedation level was determined every 3 min for 1 h, then recovery time and survival were measured. In Study 1, MS‐222 and 2‐phenoxyethanol were determined to be ineffective on prawn at all rates tested. Based on their performance in Study 1, quinaldine, clove oil, and AquiS^TM were evaluated at 100, 200, and 300 mg/L in Study 2. Observations were determined as in Study 1. Clove oil and Aqui‐S^TM induced anesthesia faster and at lower concentrations than quinaldine. At the highest treatment rate (300 mg/L) prawn suffered 60% mortality in the Aqui‐S^TM treatment, 13% mortality in the quinaldine treatment, and 0% mortality in the clove oil treatment and control following a 1‐h exposure to these concentrations. Based on these data, Aqui‐S^TM and clove oil applied at 100 mg/L may be suitable anesthetic treatments for prawn. Additional research is needed to determine optimal time and dose relationships to minimize stress during holding, handling, and transportation of prawn.     

Response to T3 treatment and changing environmental salinity of liver lipid composition, mitochondrial respiration and (Na++ ++ K++)-ATPase activity in rainbow trout Oncorhynchus mykiss Walbaum

Rainbow trout Oncorhynchus mykiss Walbaum were fed a pelleted diet (14.3% wet weight lipid) containing 9 p.p.m. 3, 5, 3′‐triiodo‐l ‐thyronine (T₃) for 1 month and then transferred from fresh water to brackish water (average 22 p.p.t. salinity), where they were maintained untreated for 22 days. Trout fed a control diet were subjected to the same protocol. For both treated and control trout, liver lipid and fatty acid composition, mitochondrial respiratory activity and oxidative phosphorylation and (Na⁺ + K⁺)‐ATPase activity were monitored in fish sampled periodically throughout the trial. No differences between treated and control trout occurred in liver total lipid, phospholipid and cholesterol content or fatty acid composition. Conversely, irrespective of T₃ administration, the trout from the two habitats showed adaptive changes to salinity, differing in phospholipids and in the fatty acid composition of total and neutral lipids and selected phospholipids. Liver membrane permeability and mitochondrial respiratory activity were affected by both T₃ treatment and salinity transfer. The latter was apparently greater than the former in affecting mitochondrial respiratory activity. The higher (Na⁺ + K⁺)‐ATPase activity in T₃‐treated trout after 22 days in brackish water may reflect long‐term effects of the hormone linked to salinity adaptation.     

Biofloc technology in intensive broodstock farming of the pink shrimp Farfantepenaeus duorarum: spawning performance,biochemical composition and fatty acid profile of eggs

A 45‐day trial was performed to evaluate the effect of biofloc technology (BFT) with or without fresh food (FF) supplementation during pre‐maturation period on Farfantepenaeus duorarum spawning performance, biochemical composition and fatty acid profile of eggs as compared with conventional clear‐water system (CW+FF). Females raised in biofloc and that received FF supplementation (FLOC+FF) achieved better spawning performance in terms of number of eggs per spawn (49 × 10³), number of eggs per spawn per g of spawner's body weight (2.1 × 10³) and egg size (~275 μm) as compared with CW+FF (23 × 10³, 1.1 × 10³ and 263 μm respectively), but both treatments did not vary from FLOC (P > 0.05). High spawning activity was also observed in biofloc system as compared with clear‐water system as shown in number of spawns per ablated female (2.2–3.0 versus 0.6) and percentage of females that spawn at least once (80–82 versus 25%). Biochemical composition of eggs presented no significant differences among treatments. FA profile of eggs indicated that high spawning activity performed by females in FLOC+FF treatment was reflected in lower mean levels of EPA, DHA and sum of polyunsaturated fatty acids (n‐3) and (n‐6). The better reproductive performance demonstrated by females raised in biofloc justified the application of this technology in F. duorarum broodstock.     

Effects of dietary propolis and vitamin E on growth performance and antioxidant status in juvenile rainbow trout (Oncorhynchus mykiss) under different flow rate

The present study investigated the effects of propolis and vitamin E supplementation in diets of juvenile rainbow trout subjected to two different flow rates (0.9 and 2.1 L min^?1) on growth performance, and vitamin A, C and E concentrations in tissues as well as malondialdehyde (MDA) levels. Juvenile rainbow trout were fed with diets containing 10 and 30 g propolis kg^?1, 60 mg kg^?1 vitamin E (Rovimix E‐50 adsorbate; min.%50 dl‐α‐tokopherly acetate) and without supplemented basal diet for 12 weeks. Weight gain (WG) in the C group was significantly lower (P < 0.05) than P10, P30 and E60 groups at both flow rate treatments. At 2.1 L min^?1, specific growth rate (SGR) in the C group was significantly lower (P < 0.05) than other groups, but at 0.9 L min^?1, SGR of fish did not differ among the diets groups (P > 0.05). Survival rate (SUR) was higher in propolis and vitamin E supplemented diet groups (P < 0.05) than control diet group at 0.9 L min^?1. Fish fed on diet E60 had higher (P < 0.05) tissue vitamin E concentration than fishes fed on other diets groups. Vitamin C concentration in rainbow trout tissues was significantly affected by the 30 g propolis supplemented diet group (P < 0.05), followed by the 10 g propolis supplemented diet group (P < 0.05). MDA level of E60 group was found significantly decreased instead of different than other groups (P < 0.05). The results of Student's t‐test revealed that WG, SGR, SUR values, vitamin (A, C, E) concentrations and MDA levels of tissues were negatively affected by 0.9 L min^?1flow rate treatment in juvenile rainbow trout.     

Impact of neem (Azadirachta indica A. Juss) seed cake upon selective nutrient cycling bacterial population in fish culture

With an objective to examine neem seed cake as bactericide, three treatments—10, 20, and 30 mg L^?1—were applied in outdoor experimental Labeo rohita culture tanks for a period of 120 days. Nitrifying, ammonifying, and denitrifying bacteria declined directly with increasing dosage. Ammonifying and phosphate solubilizing bacteria recovered within a month, nitrifying and denitrifying bacteria after 45 days. A positive relationship was established between soil available-N and ammonifying, nitrifying, and denitrifying bacteria in 10 mg L^?1 (R² = 0.68–0.83). Phosphate solubilizing bacteria responded directly with dosage and contributed significantly to the orthophosphate pool (R² = 0.66–0.78).     

In vitro assay of plant protease inhibitors from four different sources on digestive proteases of rohu, Labeo rohita (Hamilton), fingerlings

The in vitro inhibitory effect of protease inhibitors from four seed extracts (soybean, grasspea, black gram and horse gram) on digestive proteases of rohu was assessed by enzyme inhibition assay and substrate sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. High proteolytic activity was detected in the intestinal extract of rohu (Labeo rohita) fingerlings at two different pH ranges (8–8.5 and 10–11). That protein digestion occurs mainly in the alkaline condition in this fish without a stomach is evident from very high trypsin activity (0.95±0.04 benzoyl‐dl ‐arginine‐p‐nitroanilide U mg protein⁻¹) in the intestine. In case of grass pea seed, more than 50% inhibition of alkaline protease activity was recorded when the ratio of inhibitor to enzyme was 9.41 μg U⁻¹. More than 40% inhibition of protease activity was recorded in case of horse gram seed when the ratio of inhibitor to enzyme was 5.51 μg U⁻¹. Black gram at 11.0 μg U⁻¹ and soybean seed proteins at 62.75 μg U⁻¹ resulted in 50% and more than 30% inhibition of digestive protease activity in rohu fingerlings respectively. A plot of the inhibition values obtained by changing the relative concentrations of enzyme/inhibitor resulted in different dose–response curves for different protein sources. The use of substrate gel electrophoresis allowed the visualization of the aforementioned differences in inhibition. Each seed extract produced a characteristic profile of protease inhibition. It is concluded that protease inhibitors present in plant protein sources adversely affect the digestive proteases in fish and hence there is a need to eliminate/reduce the amount of such inhibitors through proper processing before incorporation into aquafeeds.     

Effects of dietary onion (Allium cepa) powder on growth performance,hemolymph indices and fillet organoleptic properties of juvenile narrow‐clawed crayfish,Astacus leptodactylus leptodactylus Eschscholtz, 1823

A 18‐week feeding trial was carried out under controlled conditions to compare the effects of onion powder (OP) at six levels (5, 10, 20, 30, 40 and 50 g/kg) on the growth performance, nutritional efficiency indices, hemolymph indices and fillet organoleptic properties of juvenile (5.62 ± 0.39 g) crayfish. The significantly (p < .05) highest values of final weight (71.30 g) and SGR (2.02% per day) and the lowest FCR (1.03) were observed in the juvenile crayfish fed the diet containing 40 g OP/kg. The juvenile crayfish fed the diet containing 40 g OP/kg had the significantly (p < .05) highest THC (105.27 × 10⁵ cell/ml), HC (98.33 × 10⁵ cell/ml), SGC (38.54 × 10⁵ cell/ml) and LGC (49.51 × 10⁵ cell/ml). The crayfish fed the levels of dietary OP higher than 30 g/kg showed the significantly (p < .05) higher values of SOD (4.07–4.30 U/min) and LYZ (6.73–7.20 U/min) compared with those fed 5, 10, 20 and 30 g of dietary OP/kg and control. Polynomial regression of SGR, FCR, PPV and PER suggested that the optimum dietary OP level could be higher than 30 and <50 mg/kg in crayfish reared in culture conditions.     

Improved techniques for rearing mud crab Scylla paramamosain (Estampador 1949) larvae

A series of rearing trials in small 1 L cones and large tanks of 30–100 L were carried out to develop optimal rearing techniques for mud crab (Scylla paramamosain) larvae. Using water exchange (discontinuous partial water renewal or continuous treatment through biofiltration) and micro‐algae (Chlorella or Chaetoceros) supplementation (daily supplementation at 0.1–0.2 million cells mL⁻¹ or maintenance at 1–2 millions cells mL⁻¹), six different types of rearing systems were tried. The combination of a green‐water batch system for early stages and a recirculating system with micro‐algae supplementation for later stages resulted in the best overall performance of the crab larvae. No clear effects of crab stocking density (50–200 larvae L⁻¹) and rotifer (30–60 rotifers mL⁻¹) and Artemia density (10–20 L⁻¹) were observed. A stocking density of 100–150 zoea 1 (Z1) L⁻¹, combined with rotifer of 30–45 mL⁻¹ for early stages and Artemia feeding at 10–15 nauplii mL⁻¹ for Z3–Z5 seemed to produce the best performance of S. paramamosain larvae. Optimal rations for crab larvae should, however, be adjusted depending on the species, larval stage, larval status, prey size, rearing system and techniques. A practical feeding schedule could be to increase live food density from 30 to 45 rotifers mL⁻¹ from Z1 to Z2 and increase the number of Artemia nauplii mL⁻¹ from 10 to 15 from Z3 to Z5. Bacterial disease remains one of the key factors underlying the high mortality in the zoea stages. Further research to develop safe prophylactic treatments is therefore warranted. Combined with proper live food enrichment techniques, application of these findings has sustained a survival rate from Z1 to crab 1–2 stages in large rearing tanks of 10–15% (maximum 30%).     

Effects of dietary propolis supplementation on growth performance, immune responses, disease resistance and body composition of juvenile eel, Anguilla japonica

This study was conducted to investigate the effects of dietary propolis supplementation on growth performance, immune responses, disease resistance and body composition of juvenile eel, Anguilla japonica, in order to evaluate its bioavailability as a feed additive for this species. A total of 540 fish averaging 7.7?±?0.22?g (mean?±?SD) were randomly distributed into 18 tanks in groups of 30, and each tank was then randomly assigned to one of three replicates of six diets containing 0 (P₀), 0.25 (P_0.25), 0.5 (P_0.5), 1.0 (P₁), 2.0 (P₂) and 4.0 (P₄) % dietary propolis. At the end of 12?weeks of feeding trial, weight gain (WG), specific growth rate (SGR), feed efficiency (FE) and protein efficiency ratio (PER) of fish fed P_0.5 diet were significantly higher than those of fish fed P₀, P₁, P₂ and P₄ diets (P?<?0.05). These parameters were 148.9%, 0.72% day^?1, 94.4% and 2.9, respectively for fish fed P_0.5 diet. Serum lysozyme activity of fish fed P_0.5 (105.7 units mL^?1) and P₁ (106.0 units mL^?1) diets were significantly higher than those of fish fed P₀, P_0.25, P₂ and P₄ diets. Mucus lysozyme activity of fish fed P₁ (8.4 units 10?cm^?2) diet was significantly higher than those of fish fed P₀, P₂ and P₄ diets. Results indicated that the optimum dietary propolis supplementation levels could be 0.25?C0.5% for optimum growth and feed efficiency, and 0.5?C1% for enhanced immune responses and disease resistance in eel, A. japonica. This study may suggest that the dietary propolis level for the optimum immune responses could be higher than the level for the optimum growth of eel.     

Contamination of palm kernel meal with Aspergillus flavus affects its nutritive value in pelleted feed for tilapia, Oreochromis mossambicus

An assessment of the nutritive value of palm kernel meal (PKM) and aflatoxin‐contaminated PKM (obtained by fermenting PKM with Aspergillus flavus) as a dietary ingredient in pelleted feed for tilapia, Oreochromis mossambicus Peters, was carried out in a 12‐week feeding trial. Seven isonitrogenous (40% crude protein) and isoenergetic (15.1 kJ g^?1) practical diets were formulated and fed close to apparent satiation to triplicate groups of 12 fish (mean initial weight 8.4 ± 0.1 g). The control diet contained 30% fish meal and 10% soybean meal (SBM) proteins. Four other experimental diets containing 20% and 50% of the SBM protein replaced by either PKM or fermented PKM, respectively, were formulated. Two additional diets containing either PKM or fermented PKM supplemented with a commercial aflatoxin adsorber (0.5% Sorbatox^TM) were also formulated. Measured aflatoxin B₁ levels in the fermented PKM‐based diets ranged from 75 to 100 µg kg^?1 diet. The growth performance and feed utilization efficiency of tilapia fed fermented PKM‐based diets were significantly lower than in fish fed the control diet at all inclusion levels (P < 0.05). Despite a small reduction, weight gains of tilapia fed PKM‐based diets were not significantly different compared with fish fed the control diet. The addition of 0.5% Sorbatox did not produce any beneficial or negative effects to the growth of tilapia. Under the dietary conditions of the present experiment, it was concluded that PKM can substitute up to 50% SBM in practical diets for O. mossambicus without much adverse effect to fish growth. However, when PKM was contaminated with A. flavus, its' incorporation into tilapia diets resulted in growth depression as a result of decreased diet digestibility and also possibly because of the presence of anti‐nutrients found in the contaminated PKM.     

Depletion of isoeugenol residues from the fillet tissue of AQUI-S™ exposed rainbow trout (Oncorhynchus mykiss)

There is a critical need in U.S. public aquaculture and fishery management for an approved sedative that allows for the immediate release of fish after sedation. AQUI-S^TM is a fish anesthetic/sedative approved for use in several countries and until recently was being developed in the U.S. as a sedative for immediate release of fish after sedation. The U.S. National Toxicology Program reported that isoeugenol (the active ingredient in AQUI-S^TM) exposed male mice showed clear evidence of carcinogenicity, therefore efforts within the U.S. Department of Interior to develop AQUI-S^TM as a sedative that allows for immediate release ceased. Despite the ruling, AQUI-S^TM still has the potential to be approved as an anesthetic with a short withdrawal time. Among the data required to gain approval for use in the U.S. are data describing the composition and depletion of all AQUI-S^TM residues from fish fillet tissue. A total residue depletion study for AQUI-S^TM was conducted by exposing market-sized rainbow trout, Oncorhynchus mykiss (mean weight, 502.7 ± 54 g; s.d.) to 8.9 mg/L of ¹⁴C-[URL]-isoeugenol for 60 min in 17 °C water. The ¹⁴C-[URL]-isoeugenol was mixed with a surfactant resulting in a mixture that mimicked AQUI-S^TM. Groups of fish (n = 6) were sampled immediately after the exposure (0-h sample group) and at 0.5, 1, 2, and 4 h thereafter. Total isoeugenol-equivalent residue concentrations in the fillet tissue were determined by oxidizing triplicate subsamples of homogenized skin-on fillet tissue from each fish to ¹⁴CO₂ and enumerating the radioactivity by static liquid scintillation counting. Isoeugenol concentrations in fillet tissue were determined by extracting homogenized fillet tissue with solvents and determining the isoeugenol concentrations in the extracts with high performance liquid chromatography techniques. The mean total isoeugenol-equivalent residue concentrations in the 0, 0.5, 1, 2, and 4-h sample groups were 55.4, 32.0, 19.8, 11.3, and 4.9 µg/g, respectively. The primary chemical residue in fillet tissue from all exposed fish was isoeugenol. The mean isoeugenol concentrations in the 0, 0.5, 1, 2, and 4-h sample groups were 48.9, 26.5, 15.3, 7.2, and 2.2 µg/g, respectively. The percents of the total radioactivity classified as isoeugenol in the 0, 0.5, 1, 2, and 4-h tissue extracts were 95, 73, 73, 64, and 48%, respectively.     

Observations on the reproductive performance of three mouth-brooding tilapia species in low-salinity underground water

The reproductive performance and seed production of three mouth‐brooding tilapia species, Oreochromis spilurus (Günther), Oreochromis aureus (Steindachner) and Taiwanese red tilapia hybrid, were compared in low‐salinity underground water (3–5 g L^?1) to select species with the highest reproductive rate. Two‐year‐old males and females having average body weights of 454 and 259 g, respectively, were stocked in 2‐m³ fibreglass breeding tanks (two tanks per species) at 1:3 male to female sex ratio and at average density of three fish m^?2. Seeds (unhatched eggs; yolk‐sac fry and the swim‐up fry) were collected weekly for 112 days. The results showed O. spilurus to have significantly higher (P<0.05) mean spawning parameters expressed as total seed tank^?1 (84 154), seed kg female^?1 day^?1 (284), seed m^?2 day^?1 (190.3) and seed female^?1 day^?1 (76.3), than O. aureus and red tilapia. Monthly seed production in O. spilurus and red tilapia gradually increased from May, peaked in July and declined in August, while in O. aureus, it continued to decrease from May to August. In this study, the observed production levels of seed in O. spilurus were among the highest levels reported for tilapia; therefore, O. spilurus could be selected as a potential candidate for mass seed production in commercial tilapia hatcheries.     

Optimizing stocking density and microalgae ration improves the growth potential of tropical black‐lip oyster,Saccostrea echinata,larvae

The combined effects of stocking density and microalgae ration on survival and size of Saccostrea echinata larvae were studied in two‐factor experiments for the major developmental stages: D‐veliger (1‐day posthatch [dph], Experiment 1), umbonate (12 dph, Experiment 2), and eyed (19 dph, Experiment 3) larvae. Larvae were stocked into replicate sets of four 10‐L aquaria with ambient 1‐μm filtered sea water (28 ± 1.5°C and 36 ppt) and cultured for four days at densities of 0.5, 2, 5, 7, or 10 larvae/mL and provided with microalgae rations at each of five densities (cells larvae^?1 day^?1); 0, 1, 3, 5, or 8 × 10³ (D‐veliger larvae, Experiment 1); 0, 5, 12, 18, or 25 × 10³ (umbonate larvae, Experiment 2); and 0, 15, 30, 40, or 60 × 10³ (eyed larvae, Experiment 3). Microalgae rations for each larval life stage were selected on the basis of increasing food requirement with larval size and comprised a 2:1:1 mixture of Chaetoceros calcitrans, Tisochrysis lutea, and Pavlova spp., calculated on an equal dry‐weight basis. Contour plots were generated from larval survival and larval size (dorso‐ventral measurement [DVM]) data to determine optimal culture conditions. Larvae showed high survival (54–100%) over a wide range of both treatment parameters across all life stages, confirming broad tolerance limits for this species. The interaction effects of larval stocking density and microalgae ration on larval size were significant (p < 0.001) across all life stages. Results indicate that maximum larval size (DVM) is achieved when S. echinata are cultured at: 6–8 larvae/mL and fed 5–6 × 10³ cells larvae^?1 day^?1 for D‐veligers (mean DVM >80 μm), at 2–8 larvae/mL and fed 11–25 × 10³ cells larvae^?1 day^?1 for umbonate larvae (mean DVM > 190 μm), and at 1–4 larvae/mL and fed 15–40 × 10³ cells larvae^?1 day^?1 for eyed larvae (mean DVM >230 μm). Results will help refine current hatchery methods for S. echinata supporting further development toward commercial aquaculture production of this species.     

Use of neem oil (Azadirachta indica) to control caligid copepod infestation on Asian seabass (Lates calcarifer)

Asian seabass (Lates calcarifer) culture in marine cages have been developed rapidly in Malaysia. The high intensity of culture might facilitate several disease infection including caligid infestation. This study was conducted to evaluate the effect of neem oil (Azadirachta indica) against caligid parasites on seabass. The prevalence and clinical pathogen of Caligus infection has been described through a sampling of 150 fish from floating cage at different cultured periods. Acute toxic tests with different concentrations of neem oil were carried out on the parasite and fish host. Results revealed that the 96‐hr median lethal concentration (LC₅₀) of Caligus and fish (10.3 ± 2.5 g of body weight) were 2 and 20 ppm respectively. The in vivo test indicated that 10 ppm of neem oil could result in 100% antiparasitic efficacy within 96 hr. These results therapeutically demonstrated the efficacy of neem oil in caligid control.     

Effects of dietary orange peel on growth performance,antioxidant activity,intestinal microbiota and liver histology of Gilthead sea bream (Sparus aurata) larvae

This study investigated the effects of dietary orange peel (OP) on growth performance, feed utilization, antioxidant activity, intestinal microbiota and liver histology of Gilthead sea bream (Sparus aurata) larvae (0.32 ± 0.01 g/fish) (mean ± SD). Fish fed iso‐nitrogenous (480 g/kg protein) and iso‐energetic (23 MJ/kg) diets supplemented with OP at concentrations of 0, 1, 3 or 5 g/kg diet, for 60 days. Growth performance and feed utilization parameters were significantly improved by the elevation of dietary OP level, and the optimum level was 5 g/kg diet. The maximum activity of the total antioxidant capacity, superoxide dismutase, glutathione peroxidase, catalase, alkaline phosphatase enzyme and malondialdehyde in the liver was found at 5.5, 4.6, 3.4, 2.9, 3.7 and 3.8 g OP/kg diet, respectively. All tested bacteria (Staphylococcus spp., Vibrio spp. and Salmonela spp) and total bacterial count decreased significantly in the gut of fish fed high levels of OP (3.0 or 5.0 g/kg). No differences were found in the liver histo‐architecture among treatments after 60 days feeding on OP diets. In summary, dietary OP improved growth rate, antioxidant activity and intestinal microbiota of S. aurata larvae with an optimum range from 2.9 to 5.5 g/kg diet.     

The in vitro effects of divalent metal ions on the activities of immune‐related enzymes in coelomic fluid from the sea cucumber Apostichopus japonicus

The immune‐related enzymes in marine animals are very sensitive to divalent metal ions. To investigate the roles divalent metal ions play in the influence on the immunity of sea cucumber Apostichopus japonicus, one of the most important commercial species in Asian countries, the effects of eight divalent metal ions at concentrations of 2.5, 5, 10, 15, 20, 25 and 30 mmol L^?1 on the activities of superoxide dismutase (SOD), phenoloxidase (PO), acid phosphatase (ACP), alkaline phosphatase (AKP) and myeloperoxidase (MPO) in coelomic fluid were determined with the nitro blue tetrazolium chloride (NBT) method, dopachrome formation method, p‐nitrophenyl phosphate (pNPP) method and 3,3′,5,5′‐tetramethyl benzidine (TMB) method. The results indicated that Mg²⁺ enhanced the activities of SOD, PO, ACP and AKP significantly and showed no obvious effect on MPO activity; Zn²⁺ increased the activities of SOD, ACP and AKP, and showed no obvious effect on the activities of PO and MPO; Cu²⁺ enhanced the activities of ACP, AKP and MPO and activated SOD and PO at a certain concentration range; Ca²⁺ and Mn²⁺ inhibited the activities of ACP and AKP; Fe²⁺ had strong inhibitory effect on SOD activity; Pb²⁺ showed inhibitions on the activities of SOD, PO, ACP and AKP; and Cd²⁺ inhibited MPO activity greatly. The data obtained in this study collectively suggest that Mg²⁺, Zn²⁺ and Cu²⁺ have potential in promotion of A. japonicus immunity, while Ca²⁺, Fe²⁺, Pb²⁺, Cd²⁺ and Mn²⁺ might be limiting factors to the immune response of A. japonicus.     

Synthesis and characterization of Azadirachta indica constructed silver nanoparticles and their immunomodulatory activity in fish

Green synthesis of nanoparticles by using different biological agents has emerged as an alternative to overcome the toxic effect of chemically synthesized nanoparticles. Among various biological agents, plants are mostly preferred. This study describes an eco‐friendly and green synthesis of silver nanoparticles (G‐AgNPs) using Azadirachta indica (neem) as a reducing agent. UV–Vis spectral analysis proved the wavelength of sample to be 420 nm, approaching the surface resonance peak specific for G‐AgNPs. Dynamic light scattering (DLS) analysis showed the mean diameter of particles as 35.4 nm with zeta potential +34.6 mV. TEM results revealed the compact and spherical shape of the particles. Fourier transform infrared spectroscopies (FT‐IR) demonstrate the presence of possible functional groups involved in synthesis of the silver nanoparticles. The functional activity of immunological parameters, such as nitroblue tetrazolium assay, myeloperoxidase activity, phagocytic activity, anti‐protease and lysozyme activity, increased significantly (P < 0.05) in fish treated with G‐AgNPs. Relative percentage survival (74%) and enhanced disease resistance were observed in G‐AgNP‐treated Cirrhinus mrigala fingerlings challenged with Aeromonas hydrophila. In summary, present results demonstrate biosynthesized silver nanoparticles have immunomodulatory and antibacterial activity.     

Ovulation and spawning induction in Caspian kutum Rutilus frisii kutum by administration of GnRHa and catecholaminergic pharmaceutical compounds with Ovaprim

The acceleration of final oocyte maturation and ovulation is recognized as a principal process in Caspian kutum Rutilus frisii kutum, a commercial and valuable species in Caspian Sea. The aim of this study was to evaluate the effects of catecholaminergic pharmaceutics which include Salbutamol (β₂‐adrenergic receptor agonist), Metoprolol (β₁‐adrenergic receptor antagonist), Clozapine and Olanzapine (the third generation of D₄ and D₂ dopamine receptor antagonists) which can be more efficient than the first and second generations, in combination with Buserelin ([D‐Ser(tBu)⁶,Pro⁹‐NEt]‐GnRHa) and Ovaprim (D‐Ala⁶,Pro⁹‐Net)‐sGnRH+Domperidone on ovulation and spawning induction in Caspian kutum. One hundred eight adult females were injected once with Buserelin Acetate (BUS) 5 μg kg^?1 BW, Ovaprim 0.5 mL kg^?1 BW, Salbutamol Sulphate (SLB) 4 mg kg^?1 BW, Olanzapine (OLZ) 5 mg kg^?1 BW, Clozapine (CZ) 12 mg kg^?1 BW and Metoprolol Tartrate (MTP) 5 mg kg^?1 BW, being divided into 12 groups: group 1, intact (negative control); group 2, Ovaprim (positive control); group 3, OLZ+SLB; group 4, OLZ+MTP; group 5, OLZ+BUS; group 6, CZ+SLB; group 7, CZ+MTP; group 8, CZ+BUS; group 9, OLZ+BUS+SLB; group 10, OLZ+BUS+MTP; group 11, CZ+BUS+SLB and group 12, CZ+BUS+MTP (N = 9). The results showed that the highest mean value in ovulation success, ovulation index, fertilization success, relative fecundity and the number of eggs belonged to Ovaprim treatment. On the other hand, spawning was successful in OLZ+BUS+SLB and CZ+BUS treatments (P < 0.05), whereas it was lower in CZ+MTP, CZ+BUS+MTP and OLZ+BUS than the other treatments. Therefore, it could be concluded that Clozapine and Olanzapine potentiated the effect of Buserelin treatment in ovulation and spawning induction, while Metoprolol blocked the stimulatory effects of GnRHa, Clozapine and Olanzapine. Salbutamol also can amplify stimulatory effect of all mentioned pharmaceutics.