Combined effects of dietary fructooligosaccharide and Bacillus licheniformis on growth performance,body composition,intestinal enzymes activities and gut histology of triangular bream (Megalobrama terminalis)

This study was conducted to investigate the effects of fructooligosaccharide (FOS) and Bacillus licheniformis (B. licheniformis) on growth performance, body composition, intestinal enzymes activities and gut histology of Megalobrama terminalis. Nine experimental diets were formulated to contain three FOS levels (0, 3 and 6 g kg^?1) and three B. licheniformis levels (0, 1 and 5 × 10⁷ CFU g^?1) following a 3 × 3 factorial design. Accordingly, diets were named as 0/0, 0/3, 0/6, 1/0, 1/3, 1/6, 5/0, 5/3 and 5/6 (B. licheniformis/FOS). At the end of the 8‐week feeding trial, weight gain (WG) and specific growth rate (SGR) of fish fed 6 g kg^?1 FOS were both significantly (P < 0.01 and P < 0.05) higher than that of the other groups in terms of dietary FOS levels. Besides, WG and SGR of fish fed 1 × 10⁷ CFU g^?1 B. licheniformis were significantly (P < 0.05) higher than that of the control group in terms of dietary B. licheniformis levels. In addition, a significant interaction (P < 0.05) between dietary FOS and B. licheniformis was observed in finial weight, WG, SGR as well as the survival rate with the highest values all observed in fish fed diet 1/3. Hepatosomatic index, carcass lipid content, lipase activities as well as microvilli length increased significantly (P < 0.05) from 0 to 1 × 10⁷ CFU g^?1, but no significant difference (P > 0.05) was observed in terms of dietary FOS levels. In addition, a significant (P < 0.05) interaction of FOS and B. licheniformis was observed in both protease and Na⁺, K⁺‐ATPase activities with the highest value obtained in fish fed diet 1/3. The results indicated that the dietary applications of dietary FOS and B. licheniformis alone or in combination can significantly improve the growth performance, survival rate, intestinal enzymes activities as well as microvilli length of triangular bream. In addition, there is a significant interaction between dietary FOS and B. licheniformis. The best combination for this species is 3 g kg^?1 FOS with 1 × 10⁷ CFU g^?1 B. licheniformis.     

Evaluation of selected metal nanoparticles on hatching and survival of larvae and fry of Indian major carp,rohu (Labeo rohita)

Different laboratory synthesized metal nanoparticles viz. Copper oxide (CuO), Zinc oxide (ZnO) and silver doped titanium dioxide (Ag‐TiO₂) were studied for their effect on hatching and survival of larvae and fry of Indian major carp, rohu, Labeo rohita both in direct application in tank water & coated onto tanks. Among these nanoparticles, CuO and ZnO nanoparticles exhibited highest percentage of hatching in both direct addition (78.0 ± 3.1% and 78.05 ± 4.2%, respectively) and coating onto tanks (58.6 ± 2.1% and 61.2 ± 2.7%, respectively) at 1 mg mL^?1 while least percentage of hatching was recorded in Ag‐TiO₂ nanoparticles irrespective of its concentration & mode of supplementation. Highest survival of L. rohita fry (50.13 ± 2.2%) was observed after 15 days post hatching in CuO coated tanks followed by ZnO coated tanks (38.6 ± 2.8%) while least was recorded in Ag‐TiO₂ coated tanks (22.53 ± 3.0%). However in control tanks coated with Poly‐Urethane base with hardener and uncoated control tanks, the survival was 42.4 ± 1.2% and 41.36 ± 1.8% respectively. Further, significantly lower microbial load of water was recorded in CuO nanoparticles coated tanks (1.5 × 10¹⁰ CFU L^?1) as compared to uncoated control tanks (1.1 × 10¹⁶ CFU L^?1) without affecting water quality parameters. On the other hand, in Ag‐TiO₂ coated tanks, significantly lower microbial load (1.0 × 10⁶ CFU L^?1) as compared to uncoated control tanks at 15 days post hatching was recorded. However, Ag‐TiO₂ was toxic to L. rohita larvae & fry both in direct application and coating onto tanks. Considering the beneficial effects of CuO nanoparticle application, it has the scope of being used in a more eco‐friendly way in hatchery operations.     

Dietary selenium requirement for on‐growing gibel carp (Carassius auratus gibelio var. CAS III)

A 12‐week growth trial was conducted in a flow‐through system to determine dietary selenium (Se) requirement for on‐growing gibel carp (initial body weight: 76.2 ± 0.05 g, mean ± SEM). Selenomethionine was supplemented to the basal diet to formulate seven semi‐purified diets containing 0.26, 0.58, 0.72, 1.14, 1.34, 1.73 and 2.09 mg Se kg^?1 diet. The results showed that plasma superoxide dismutase (SOD) activity significantly increased when fish were fed with 0.58 mg Se kg^?1 diet (P < 0.05) and then decreased at 2.09 mg Se kg^?1 diet (P < 0.05). Plasma T‐AOC activity was higher in fish fed with 0.72 mg Se kg^?1 diet (P < 0.05) and plasma malondialdehyde (MDA) was higher in fish fed with 0.26 mg Se kg^?1 diet (P < 0.05). When fish were fed 1.14 mg Se kg^?1 diet, hepatic GSH‐Px, T‐AOC, GSH and CAT activities were significantly higher than those fed with 0.26 mg Se kg^?1 diet (P < 0.05). Hepatic superoxide dismutase (SOD) activity was higher at 1.34 mg Se kg^?1 diet (P < 0.05). Fish liver Se concentrations were significantly higher when fed with 0.72 mg Se kg^?1 diet (P < 0.05) and then kept constant when Se ≥ 0.72 mg kg^?1 (P > 0.05). Whole‐body and muscle Se concentrations were higher when fed with 1.34 mg Se kg^?1 diet (P < 0.05) and kept a plateau when Se ≥ 1.34 mg kg^?1 (P > 0.05). In conclusion, based on broken‐line regression of hepatic Se concentrations, hepatic SOD activity and hepatic T‐AOC activity, dietary Se requirements for on‐growing gibel carp was 0.73 mg kg^?1, 1.12 mg kg^?1 and 1.19 mg kg^?1 diet respectively.     

Interactions of different sodium and potassium concentrations on Macrobrachium rosenbergii (de Man) offspring quality parameters

The present study evaluated various sodium and potassium concentrations in hatchery water to determine which proportions would be optimal for Macrobrachium rosenbergii larviculture. Using a closed RAS system (60‐L), experiments were conducted in two stages. In the first stage, larval quality parameters were compared among triplicate treatments of sodium (2000, 3000, 4000 and 5000 mg L^?1) and potassium (100, 150, 200 and 250 mg L^?1). During the second stage, these same parameters were compared from interactions of the two best concentrations determined in the first stage. Initial larval density was fixed at 100 larvae L^?1 and larval quality parameters such as larval stage index (LSI), larval condition index (LCI), larvae dry weight, survival (%), LC₅₀‐24 h for formalin stress and time of the first postlarvae (PL) appearance were measured. Results showed that during the early larval period time LSI, LCI and survival parameters were affected only by potassium and the interaction with sodium was not significant. At a later period of the larval development, interactions between both sodium and potassium were measurable for LSI (P < 0.05) while the interactions on LCI and survival were not significant. Measurable differences among the combined treatments 4000 mg L^?1 sodium and 150 mg L^?1 potassium resulted in the best performance for M. rosenbergii larviculture. This concentration also provided the highest final survival to PL metamorphosis (40.6 ± 2.5%) which was at least 10% higher than the other treatments.     

Effectiveness of copper sulphate,potassium permanganate and peracetic acid to reduce mortality and infestation of Ichthyobodo necator in channel catfish Ictalurus punctatus (Rafinesque 1818)

Ichthyobodo necator is a single‐celled biflagellate parasite, which in high density can cause significant mortality in young fish. Copper sulphate (CuSO₄), potassium permanganate (KMnO₄) and peracetic acid (PAA) were evaluated for effectiveness against ichthyobodosis. Treatments were: untreated control, 2.1 mg L^?1CuSO₄, 3.0 mg L^?1 KMnO₄, 1.5 mg L^?1 PAA and 3.0 mg L^?1 PAA, and were applied to flow‐through tanks on three consecutive days. The study was designed to simulate the flow‐through systems utilized in the commercial rearing of juvenile channel catfish (Ictalurus punctatus). Mortality was monitored daily to compare survival rate among treatments. Parasite intensity was assessed pre chemical exposure and 20–24 h after the third application to determine effectiveness of the treatment. An assessment was also done 7 days post application to investigate possible reoccurrence. Copper sulphate, KMnO₄ and PAA (3.0 mg L^?1) significantly reduced the infestation rate of I. necator. Copper sulphate significantly improved the survival of I. necator infested channel catfish after three flow‐through applications compared with the untreated control. The 3.0 mg L^?1 PAA resulted in significantly lower survival than the untreated control, the 1.5 mg L^?1 PAA and the KMnO₄ were not statistically different from the untreated control.     

Acute toxicity of nitrite on white shrimp Litopenaeus vannamei (Boone) juveniles in low‐salinity water

The nitrite toxicity was estimated in juveniles of L. vannamei. The 24, 48, 72 and 96 h LC₅₀ of nitrite‐N on juveniles were 8.1, 7.9, 6.8 and 5.7 mg L^?1 at 0.6 g L^?1; 14.4, 9.6 8.3 and 7.0 mg L^?1 at 1.0 g L^?1; 19.4, 15.4, 13.4 and 12.4 mg L^?1 at 2.0 g L^?1 of salinity respectively. The tolerance of juveniles to nitrite decreased at 96 h of exposure by 18.6% and 54.0%, when salinity declined from 1.0 to 0.6 g L^?1 and from 2.0 to 0.6 g L^?1 respectively. The safe concentrations at salinities of 0.6, 1.0 and 2.0 g L^?1 were 0.28, 0.35 and 0.62 mg L^?1 nitrite‐N respectively. The relationship between LC₅₀ (mg L^?1), salinity (S) (g L^?1) and exposure time (T) (h) was LC₅₀ = 8.4688 + 5.6764S – 0.0762T for salinities from 0.6 to 2.0 g L^?1 and for exposure times from 24 to 96 h; the relationship between survival (%) and nitrite‐N concentration (C) for salinity of 0.6–2.0 g L^?1, nitrite‐N concentrations of 0–40 mg L^?1 and exposure times from 0 to 96 h was as follows: survival (%) = 0.8442 + 0.1909S – 0.0038T – 0.0277C + 0.0008ST + 0.0001CT–0.0029SC, and the tentative equation for predicting the 96‐h LC₅₀ to salinities from 0.6 to 35 g L^?1 in L. vannamei juveniles (3.9–4.4 g) was 96‐h LC₅₀ = 0.2127 S² + 1.558S + 5.9868. For nitrite toxicity, it is shown that a small change in salinity of waters from 2.0 to 0.6 g L^?1 is more critical for L. vannamei than when wider differences in salinity occur in brackish and marine waters (15–35 g L^?1).     

Determination of histological and genotoxic effects of formalin on Nile tilapia (Oreochromis niloticus L.)

Formalin (37–40% formaldehyde) is one of the most effective, widely used chemical in pisciculture for its antiparasitic, antifungal and prophylactic activities. It is used in paints, cleaning products and textile industry, as well. Genotoxic and histological effects of sublethal formalin exposure on Nile tilapia (Oreochromis niloticus) are investigated in this study. The semi‐static acute test was employed. One‐tenth, one‐fifth and half of the 96‐h LC₅₀ value, 148 mg L^?1, were used as sublethal exposure concentrations: 15, 30 and 75 mg L^?1 for 24, 48, 96 and 168 h respectively. The micronucleus test was applied to investigate the genotoxic effects on fish erythrocytes at the end of predetermined exposure periods and all tissues of the fish were sampled for histological examination. The micronucleus frequencies increased significantly in all exposure groups when compared with their control groups in a dose and time dependent manner (P < 0.05). Results of histological examination showed no histopathological findings in the gonads, heart, skeletal muscle, spleen and intestine tissues after sublethal formalin exposure. Passive hyperaemia of liver tissue in varying proportions, hydropic degeneration, fatty degeneration in high concentrations, branchitis on gill tissues in varying proportions, epithelial lifting, telangiectasia, bleeding in kidney and hyperaemia were found in Nile tilapia exposed to formalin at different concentrations and durations. As a result, it is thought that formalin leads to tissue damage and shows genotoxic effects even at 15 mg L^?1 concentration in O. niloticus.     

In vitro and in vivo efficacy of drugs against the protozoan parasite Azumiobodo hoyamushi that causes soft tunic syndrome in the edible ascidian Halocynthia roretzi (Drasche)

It was discovered recently that infection by a protozoan parasite, Azumiobodo hoyamushi, is the most probable cause for soft tunic syndrome in an edible ascidian, Halocynthia roretzi (Drasche). In an attempt to develop measures to eradicate the causative parasite, various drugs were tested for efficacy in vitro and in vivo. Of the 20 antiprotozoal drugs having different action mechanisms, five were found potent (24‐h EC₅₀ < 10 mg L^?1) in their parasite‐killing effects: formalin, H₂O₂, bithionol, ClO₂ and bronopol. Moderately potent drugs (10 < 24‐h EC₅₀ < 100 mg L^?1) were quinine, fumagillin, amphotericin B, ketoconazole, povidone‐iodine, chloramine‐T and benzalkonium chloride. Seven compounds, metronidazole, albendazole, paromomycin, nalidixic acid, sulfamonomethoxine, KMnO₄, potassium monopersulphate and citric acid, exhibited EC₅₀ > 100 mg L^?1. When ascidians were artificially infected with A. hoyamushi, treated using 40 mg L^?1 formalin, bronopol, ClO₂, or H₂O₂ for 1 h and then monitored for 24 h, very low mortality was observed. However, the number of surviving parasite cells in the ascidian tunic tissues was significantly reduced by treating with 40 mg L^?1 formalin or ClO₂ for 1 h. The data suggest that we might be able to develop a disinfection measure using a treatment regimen involving commonly available drugs.     

Acute toxicity of ammonia in Meagre (Argyrosomus regius Asso, 1801) at different temperatures

Argyrosomus regius (3.0 ± 0.9 g) were exposed to different concentrations of ammonia in a series of acute toxicity tests by the static renewal method at three temperature levels (18, 22 and 26°C) at a pH of 8.2. Low temperature clearly increased the tolerance of the fish to total ammonia nitrogen (TAN) and unionized ammonia (NH₃) (P < 0.05). While the 96‐h LC₅₀ values of TAN were 19.79, 10.39 and 5.06 mg L^?1, the 96‐h LC₅₀ of NH₃ were 1.00, 0.70 and 0.44 mg L^?1 at 18, 22 and 26°C respectively. The safe levels of NH₃ for A. regius was estimated to be 0.10, 0.07 and 0.04 mg L^?1 at 18, 22 and 26°C respectively (P < 0.05). This study clearly indicates that A. regius is more sensitive to ammonia than other marine fish species cultured on the Mediterranean and Eastern Atlantic coasts.     

Survival and respiration of marbled rabbitfish (Siganus rivulatus) fingerlings at various oxygen tensions

Marbled rabbitfish, Siganus rivulatus, is an economically valuable herbivorous fish and a potential candidate for warmwater aquaculture. This study was carried out to: (1) assess the effect of various oxygen concentrations on survival and behaviour of S. rivulatus fingerlings and (2) investigate the response of S. rivulatus to hypoxia and determine its critical oxygen tension (P_crit). In the first experiment, groups of rabbitfish (15 fish per group) were maintained for 1 h in waters of various oxygen concentrations. They were then transferred to well‐aerated tanks and observed for 72 h. Survival was recorded, fish behaviour at low oxygen concentrations observed, and LC₅₀ after 1‐h hypoxia and 72‐h recovery evaluated. In the second experiment, a series of stop‐flow respirometry experiments were performed during which dissolved oxygen was allowed to drop to 0.5 mg L^?1 and respiration rate recorded at various oxygen concentrations. In the first experiment, all fish survived for 1 h at oxygen concentration of 1.44 mg L^?1 and greater, but started dying at oxygen concentrations below 0.65 mg L^?1 (16% survival). The LC₅₀ of S. rivulatus fingerlings was 0.6 mg L^?1. Results of the second experiment showed that S. rivulatus is an oxyregulator until P_crit (1.7 mg L^?1 O₂) is reached, becoming an oxyconformer below this concentration. Findings allow for a better understanding of environmental oxygen tolerances and minimum acceptable oxygen concentration in rabbitfish aquaculture.     

Effect of florfenicol and oxytetracycline treatments on the intensive larval culture of the Chilean scallop Argopecten purpuratus (Lamarck, 1819)

The administration of antimicrobials to control bacterial pathologies in Chilean scallop hatcheries is a frequent practice, but their effects on these cultures remained unknown. This study was undertaken to obtain information on the effect of the administration of florfenicol and oxytetracycline on the growth, survival and bacterial content of scallop larvae under farming conditions. Florfenicol‐treated cultures exhibited high survival rates (44% after 17 days of culture), whereas cultures not treated or treated with oxytetracycline collapsed after 11 days of culture. Surprisingly, no significant differences in the heterotrophic (Tukey test; P = 0.226) and Vibrio (Tukey test; P = 0.666) concentrations between the oxytetracycline‐treated and untreated larval cultures were observed. Otherwise, florfenicol administered directly into rearing tanks produced significantly higher larval growth (Tukey test; P = 0.0001) and survival (Tukey test; P = 0.011) than bath treatment. When 2 and 4 mg L^?1 of florfenicol were compared, no significant differences in growth (t‐test; P = 0.4596) and survival (Tukey test; P = 0.057) were observed, suggesting that a concentration of 2 mg L^?1 is sufficient to ensure larval production. The present results demonstrate the efficacy of florfenicol‐based therapy to increase larval survival and growth at commercial scale and prompt the necessity to standardize its use in Chilean scallop hatcheries.     

Acute toxicity of ammonia in Pacu fish (Piaractus mesopotamicus,Holmberg, 1887) at different temperatures levels

Piaractus mesopotamicus juveniles (total length 12 ± 0.5 mm) were exposed to different concentrations of ammonia‐N (un‐ionized plus ionized ammonia as nitrogen), using the static renewal method at different temperature levels (15, 20 and 25°C) at pH 7. The 24, 48, 72, 96 h LC₅₀ values of ammonia‐N in P. mesopotamicus juveniles were 5.32, 4.19, 3.79 and 2.85 mg L^?1 at 15°C; 4.81, 3.97, 3.25 and 2.50 mg L^?1 at 20°C; and 4.16, 3.79, 2.58 and 1.97 mg L^?1 at 25°C respectively. The 24, 48, 72, 96 h LC₅₀ values of NH₃‐N (un‐ionized ammonia as nitrogen) were 0.018, 0.014, 0.013, 0.009 mg L^?1 at 15°C temperature; 0.023, 0.019, 0.016 and 0.012 mg L^?1 at 20°C; 0.029, 0.026, 0.018 and 0.014 mg L^?1 at 25°C. The temperature increase from 15 to 25°C caused an increase of ammonia‐N susceptibility by 21.80%, 9.55%, 31.92% and 30.87%, after 24, 48, 72 and 96 h exposure respectively. Furthermore, we found that exposure of fish to ammonia‐N caused an elevation in total haemoglobin and blood glucose with an increase of 2 mg L^?1 concentration. Ammonia levels tolerated, especially in different temperatures levels, have important implications for the management of aquaculture.     

Effects of dietary ethoxyquin on growth,feed utilization and residue in the muscle of juvenile Japanese seabass,Lateolabrax japonicus

Ethoxyquin (EQ) is the most common synthetic antioxidant used for preventing rancidity in fish foodstuffs. However, literature related to the effects of dietary EQ on performance of fish was limited. The present study was conducted to investigate the effects of EQ on performance and EQ residue in muscle of juvenile Japanese seabass Lateolabrax japonicus and to estimate the optimal EQ concentration in the diet. Graded levels [0 (control), 50, 150, 450 and 1350 mg EQ kg^?1 diet] of EQ were added to the basal diet, resulting in five dietary treatments in the experiment. Each diet was fed to triplicate groups of seabass (initial body weight 8.01 ± 0.76 g) for 12 weeks in floating sea cages (1.5 × 1.5 × 2.0 m, 30 fish per cage). Survival ranged from 78.9 to 86.7%, and was irrespective of dietary EQ levels. The specific growth rate (SGR) of fish fed diets supplemented with ≤50 mg kg^?1 EQ had significantly (P < 0.05) higher SGR than fish fed diets supplemented with ≥150 mg kg^?1 EQ, the highest SGR was observed in fish fed diet with 50 mg kg^?1 EQ supplementation. Feed intake (FI) and feed efficiency (FE) were not significantly (P > 0.05) different among dietary treatments. Fish fed diets with 50 and 1350 mg kg^?1 EQ had a significant (P < 0.05) lower body lipid content than fish in the control group. Muscle EQ level significantly increased when dietary EQ increased. Optimal EQ concentration estimated by polynomial regression based on maximum growth of juvenile Japanese seabass was 13.78 mg kg^?1 diet.     

Efficacy of calcein as a chemical marker of green‐lipped mussel (Perna canaliculus) larvae and its potential use for tracking larval dispersal

An optimal chemical shell marking protocol was developed for the New Zealand green‐lipped mussel, Perna canaliculus with a view to its future use in larval tracking experiments. Larval P. canaliculus aged either 10, 15 or 19 days post fertilization were immersed in treatments of 50, 100 and 200 mg L^?1 of calcein for a period of 24 h before measurements of shell mark brightness were taken. There was 100% marking success in all calcein treatments for all age classes, with 19‐day larvae immersed in 200 mg L^?1 calcein producing the brightest mark. Growth was not affected by calcein immersion; however, 10‐day larvae exhibited significantly higher levels of mortality compared with 15‐ and 19‐day larvae suggesting a reduced resilience to the marking protocols in younger larvae. In a mass staining experiment, a solution of 100 mg L^?1 calcein was used to successfully stain15.6 million hatchery reared P. canaliculus larvae. Calcein, therefore, offers a low impact method with which to stain the sensitive early life stages of this species thus providing a rapid method for identifying individuals of interest, i.e. individuals released in the wild or specific family lines within a hatchery environment.     

Hen egg yolk and skinned krill as possible foods for rearing leptocephalus larvae of Anguilla japonica Temminck & Schlegel

Usual diets for rearing leptocephalus larvae of Japanese eel Anguilla japonica include eggs of the endangered spiny dogfish Squalus acanthias (SE). We investigated the effects of alternative food materials, hen egg yolk (HEY) and exoskeleton‐free (skinned) Antarctic krill (SAK), on the growth and survival of eel larvae. We found that feed comprising whole krill including exoskeleton (WAK) containing higher levels of fluoride (37.89 mg kg^?1) was acutely toxic to eel larvae exposed to this alone. In contrast, extract from SAK containing lower concentrations of fluoride (4.25 mg kg^?1) showed no apparent adverse effects. Growth of larvae fed a mixture of SE and SAK in a feed trial of 58 days [mean body weight (BW), 6.0 mg] was about twofold higher than that of larvae fed a mixture of SE and WAK (3.2 mg) (P < 0.01). A mixture of HEY and SAK also had some dietary benefits for eel larvae, enabling them to survive for up to 58 days and to grow significantly (mean BW, 2.4 mg), compared with their initial weight (mean BW, 0.2 mg) (P < 0.001). Although additional nutritional improvements are needed, the present results suggest that combination diet HEY and SAK may be a good alternative to SE as an effective diet for eel larvae.     

Studies on bacterial pathogens isolated from diseased torafugu (Takifugu rubripes) cultured in marine industrial recirculation aquaculture system in Shandong Province,China

In spring of 2011, an epidemic outbreak of torafugu with high mortality occurred in an aquafarm with marine industrial recirculation aquaculture system (MIRAS) in Yantai, Shandong Province, China. The diseased fish showed anorexia, haemorrhaging and festering fin and skin and swelling internal organs. Forty‐five dominant bacterial isolates were obtained from the diseased fish, and were found to belong to 12 species according to 16S rRNA gene sequences. One strain from each species was selected to test the pathogenicity, and five strains were showed to be virulent to zebrafish. Whereas Enterovibrio nigricans Fr42 was highly virulent with the LD₅₀ of 7.8 × 10⁴ CFU g^?1, Photobacterium swingsii Fr23, Vibrio owensii Fr40, V. harveyi Fr51 and V. rotiferianus Fr71 were moderately virulent with the LD₅₀ of 1.7 × 10⁶ to 8.4 × 10⁶ CFU g^?1. Both the bacteria and their extracellular products of the five strains were found to show phospholipase, caseinase, gelatinase, amylase and/or lipase activities. The production of N‐acyl homoserine lactones (AHLs) of the five strains was detected by three different AHLs biosensors, and three of them were found to produce AHLs by at least one kind of biosensor. This is the first study describing various opportunistic bacterial pathogens of fish cultured in MIRAS in China.     

Ammonia tolerance of Litopenaeus vannamei (Boone) larvae

The tolerance of Litopenaeus vannamei larvae to increasing concentrations of total ammonia nitrogen (TAN) using a short‐term static renewal method at 26°C, 34 g L^?1 salinity and pH 8.5 was assessed. The median lethal concentration (24 h LC₅₀) for TAN in zoea (1‐2‐3), mysis (1‐2‐3) and postlarvae 1 were, respectively, 4.2‐9.9‐16.0; 19.0‐17.3‐17.5 and 13.2 mg L^?1TAN (0.6‐1.5‐2.4; 2.8‐2.5‐2.6 and 1.9 mg L^?1 NH₃‐N). The LC₅₀ values obtained in this study suggest that zoeal and post‐larval stages are more sensitive to 24 h ammonia exposure than the mysis stage of L. vannamei larvae. On the basis of the ammonia toxicity level (24 h LC₅₀) at zoea 1, we recommend that this level does not exceed 0.42 mg L^?1 TAN – equivalent to 0.06 mg L^?1 NH₃‐N – to reduce ammonia toxicity during the rearing of L. vannamei larvae.     

Effect of ammonia on the electrolyte status of an Indian major carp Catla catla

Ammonia is present in the aquatic environment due to agricultural runoff, sewage effluents and decomposition of biological waste. High level of ammonium in the aquatic environment or in aquaculture farms is a serious problem for aquatic organisms. In the present investigation, the sublethal effects of total ammonia nitrogen (TAN) at 9.98 mg L^?1 on plasma electrolytes (Na⁺, K⁺ and Cl^?) level of Indian major carp Catla catla fingerlings was studied for a period of 35 days under semistatic conditions. Exposure of fish to sublethal concentration of TAN at 9.98 mg L^?1 caused changes in the levels of plasma electrolytes (Na⁺, K⁺ and Cl^?) compared with control fish. No mortality was observed in any group during the treatment period. The present study reveals that the levels of plasma sodium and potassium (except on 7th day) electrolytes were significantly (P < 0.05) decreased throughout the study period. However, the decrease in plasma chloride level was found to be not significant when compared with control group. The present study concludes that the measurements of plasma electrolytes levels can be used as potential biomarkers for monitoring nitrogenous chemicals in aquatic environment.     

Bioassay‐guided isolation and identification of active compounds from Macleaya microcarpa (Maxim) Fedde against fish pathogenic bacteria

Four alkaloids (Sanguinarine, 6‐Methoxyl‐dihydro‐chelerythrine, Cryptopine and β‐Allocryptopine) were isolated from aerial parts of Macleaya microcarpa (Maxim) Fedde using bioassay‐guided isolation method, and the inhibitory activity of ethanolic extract, various fractions and these four alkaloids against four fish pathogenic bacteria (Aeromonas hydrophila, Aeromonas salmonicida, Vibrio anguillarum and Vibrio harveyi) was assessed in vitro using the agar dilution method and the microdilution assay method respectively. A. hydrophila was the most sensitive strain to all the tested compounds. Minimum inhibitory concentration (MIC) values were lower for sanguinarine against all tested Gram‐negative strains than other three alkaloids, with MIC values of 12.5 mg L^?1 for A. hydrophila and 50 mg L^?1 to other pathogenic bacteria. Followed by 6‐methoxyl‐dihydro‐chelerythrine, which showed considerable antibacterial activity with MIC values of 80 mg L^?1 for A. hydrophila, 100 mg L^?1 for V. harveyi, and 125 mg L^?1 for both V. anguillarum and A. salmonicida. Cryptopine and β‐allocryptopine revealed similar inhibitory activity with MIC values of 100 mg L^?1 for A. hydrophila and 200 mg L^?1 for other three bacterial species. These finding provided evidence that extract, as well as isolated compounds from M. microcarpa might be potential sources novel antibacterial agents for the treatment of fish infectious diseases.     

Growth,digestive and absorptive abilities and antioxidative capacity in the hepatopancreas and intestine of young grass carp (Ctenopharyngodon idellus Val.) fed graded levels of dietary manganese

To study the effects of manganese on growth performance, digestive and absorptive abilities, as well as the antioxidative capacity in the hepatopancreas and intestine, young grass carp (Ctenopharyngodon idellus Val.) (264 ± 1 g) were fed diets containing graded levels of manganese at 3.65 (control), 8.62, 13.48, 18.24, 22.97 and 27.86 mg kg^?1 diet for 8 weeks. Per cent weight gain (PWG) and feed intake were the poorest in fish fed the basal diet (P < 0.05). The activities of trypsin, lipase and alkaline phosphatase in the intestine were significantly enhanced with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05). Additionally, in the hepatopancreas and intestine, the protein carbonyl and malondialdehyde contents were the lowest in fish fed the diet with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05), while the anti‐hydroxyl radical capacities, manganese superoxide dismutase (MnSOD), glutathione peroxidase and glutathione‐S‐transferase activities were significantly enhanced with dietary manganese level at 13.48 mg kg^?1 diet (P < 0.05). Moreover, the catalase activity and glutathione content in the intestine were the highest in fish fed the diet with dietary manganese level at 18.24 mg kg^?1 diet (P < 0.05). These results indicated that optimum dietary manganese promoted growth, enhanced the digestive and absorptive abilities, and improved the antioxidative capacity in young grass carp. Based on the quadratic regression analysis for PWG and intestinal MnSOD activity, the manganese requirements for young grass carp with the initial body weight of 264 g were 16.91 and 18.21 mg kg^?1 diet respectively.