The effect of banana blossom on growth performance, immune response, disease resistance, and anti-hypothermal stress of Macrobrachium rosenbergii.

Banana (Musa acuminata) blossom contains high nutritional value and bioactive compounds. In this study, Macrobrachium rosenbergii were fed with diets containing banana blossom powder (BBP) at 10 and 20 g kg-1, hot-banana blossom (BBH) extract at 10 and 20 g kg-1, and the basal diet for 56 days. The growth performance, physiological response and immune parameters were evaluated. The results showed that a significantly higher percentage weight gain (PWG) and percentage length gain (PLG) in prawns fed with BBH diet. The feed efficiency (FE) significantly increased in prawns fed BBP. The prawn fed both BBH and BBP diet showed higher survival rate than control group. The prawn fed with BBH showed a significant increase in total haemocyte count (THC) and different haemocyte count (DHC), whereas phenoloxidase (PO) activity and respiratory bursts (RBs) significant increase in prawns fed both BBP and BBH diet. Furthermore, M. rosenbergii fed with both BBP and BBH diets showed significantly higher phagocytic activity and clearance efficiency against Lactococcus garvieae infection. At the end of the 56 days of feeding trial, the susceptibility of prawns to L. garvieae infection and hypothermal (18 °C) stress were evaluated. The results showed that prawns fed BBH diets had a significantly higher survival rate against L. garvieae than those of fed with the basal diet. Anti-hypothermal stress was observed in prawns fed both BBP and BBH diets showing no significant difference in haemolymph glucose in prawns subjected to 18 °C and 28 °C, whereas the norepinephrine level in haemolymph of prawns fed with BBH diets subjected to 18 °C was significantly lower than in prawns subjected to 28 °C. In summary, we recommend addition of hot-banana blossom extract to the diet of M. rosenbergii at 20 g kg-1 to promote growth performance, improve physiological function, enhance immunity, increase anti-hypothermal stress, and to increase resistance against L. gavieae.

Effect of Cymbopogon citratus Essential Oil (EO) on Handling Stress in Giant Freshwater Prawn (Macrobrachium rosenbergii).

BACKGROUND AND OBJECTIVE: Effects of Cymbopogon citratus essential oil (EO) was tested on minimizing handling stress in Macrobrachium rosenbergii through the evaluation of their metabolite responses [glucose, lactate, glycogen, protein, Lactate Dehydrogenase (LDH), Malate Dehydrogenase (MDH), Acetylcholinesterase (AChE) and Alanine Aminotransferase (ALT)]. This study aimed to investigate the efficacy of C. citratus extract in the anaesthetization of M. rosenbergii. MATERIALS AND METHODS: Three treatments including control, prawn exposed to stress alone (T1) and prawn exposed to stress in the presence of C. citratus EO (T2) were tested. A C. citratus EO at 500 µL L-1 had been determined in a previous study and was selected as the critical dose to be applied as an anesthetic agent. Handling stress was induced into prawns by netting, at 2 min interval for 30 min and their hemolymph were collected to determine the metabolite responses. RESULTS: The increase of glucose, lactate and LDH of M. rosenbergii when exposed to handling stress alone (T1) in comparison to T2 (stress with anesthetic C. citratus EO) were identified. Further, a low glycogen level in parallel with low AChE activity was observed which indicates the involvement of secondary metabolites to cope with the energy demand in T1 over T2. CONCLUSION: This study indicates the efficiency of C. citratus EO to reduce stress during handling in M. rosenbergii.

Modulation of antioxidant gene expressions by Roundup® exposure in the decapod Macrobrachium potiuna.

Glyphosate-based herbicides (GBH), including Roundup®, are the most used herbicides in agricultural and non-agricultural areas, which can reach aquatic environments through drift during application or surface runoff. Some studies, mostly in fish, demonstrated that GBH caused oxidative stress in non-target animals. However, only few information is available on the GBH effects in the antioxidant and stress proteins of many other organisms, such as freshwater crustaceans. Thus, we aimed to investigate the effects of environmentally relevant GBH concentrations on the relative transcript expression (RTE) of the superoxide dismutase (sod1), catalase (cat), selenium-dependent glutathione peroxidase (gpx), glutathione-S-transferase (gst), thioredoxin (txn), heat shock protein (hsp70 and hsp90) in the hepatopancreas of the ecologically important freshwater prawn Macrobrachium potiuna. Moreover, this study aimed to assess the gender-differences responses to GBH exposure. Male and female prawns were exposed to three Roundup WG® concentrations (0.0065, 0.065 and 0.28 mg of glyphosate/L) and a control group (0.0 mg/L) for 7 and 14 days. In general, males had an under-expression of the studied genes, indicating an oxidative stress and possible accumulation of ROS in the hepatopancreas. In the opposite, females had an overexpression of the same genes, indicating a more robust antioxidant system, in order to cope with the possible ROS increase after Roundup WG® exposure. Therefore, results confirmed that gender could be a confounding factor in ecotoxicological assessment of GBH effects. Additionally, this work highlights that sod1, cat, gpx, gst, txn, hsp70 and hsp90 gene expressions seem to be useful biomarkers to investigate the oxidative stress caused by Roundup WG® in Macrobrachium sp.

Effects of Moringa oleifera leaf extract on growth performance, physiological and immune response, and related immune gene expression of Macrobrachium rosenbergii with Vibrio anguillarum and ammonia stress.

In order to study the effects of Moringa oleifera leaf extract on Macrobrachium rosenbergii under high ammonia exposure, freshwater prawns were randomly divided into five groups: a control group was fed with basal diet, and four treatment groups fed with basal diet supplemented with 0.25%, 0.5% and 1.0% M. oleifera leaf extract and 0.025% Enrofloxacin for 60 days, respectively. Then, freshwater prawns were exposed to high ammonia stress for 72 h and Vibro anguillarum infection. The growth, antioxidant capabilities, related immune genes as well as resistance to infection by V. anguillarum were determined. The results showed that compared with the control group, the weight gain, specific growth rate and protein efficiency rate, haemolymph catalase (CAT), superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) increased while feed conversion ratio, haemolymph aspartate aminotransferase, alanine aminotransferase, nitrogen oxide (NO), hepatopancreas heat shock proteins (HSP70), immune deficiency (IMD) expression levels decreased in the group of 0.5% M. oleifera leaf extract before the stress. After ammonia stress, the group of 0.5% M. oleifera leaf extract also could improve the haemolymph SOD, glutathione peroxidase, NO, iNOS, hepatopancreas HSP70 expression levels and reduce haemolymph CAT, hepatopancreas peroxiredoxin 5 and NF kappa B inhibitor alpha expression level compared with the control group. The rate of mortality of the prawns challenged with V. anguillarum was lower in the supplemented groups in comparison with the control group with the lowest being in the group of 0.5% M. oleifera leaf extract. Antioxidant activities as well as biochemical parameters in the enrofloxacin group (0.025%E) were not significantly enhanced both pre and post challenge in comparison with the M. oleifera leaf extract groups, showing the superiority of the natural herb over the synthetic antibiotic. In summary, this study suggested that at an inclusion rate of 0.5%, M. oleifera leaf extract could increase the growth performance, even has positive effects on physiological and immune function and prevents high ammonia stress in the Freshwater prawn, M.rosenbergii.

Toxicity comparison of the shoreline cleaners Accell Clean® and PES-51® in two life stages of the grass shrimp, Palaemonetes pugio.

Oil spills are a significant source of coastal pollution. Shoreline cleaners, used to remove oil from surfaces during spill response and remediation, may also act as toxins. Adult and larval grass shrimp, Palaemonetes pugio, were tested for lethal and sublethal impacts from two shoreline cleaners, Accell Clean SWA® and PES-51®, alone and in combination with crude oil using Chemically Enhanced Water Accommodated Fractions (CEWAFs). Median lethal toxicity values determined for the individual cleaners were similar. However, when tested in mixture with oil as CEWAFs, Accell Clean SWA resulted in greater hydrocarbon concentrations in the water column and greater toxicity than PES-51. Increased glutathione levels were observed for adult shrimp exposed to Accell Clean SWA, and glutathione was elevated in shrimp exposed to both CEWAFs. Larval shrimp development was delayed after exposure to both CEWAFs. These findings may have implications for managing and mitigating oil spills.

Effect of microencapsulated probiotic Bacillus vireti 01-polysaccharide extract of Gracilaria folifera with alginate-chitosan on immunity, antioxidant activity and disease resistance of Macrobrachium rosenbergii against Aeromonas hydrophila infection.

Polysaccharide from red seaweed Gracilaria folifera has an interesting functional property of antioxidant activity and prebiotic effect. A feeding trial experiment was directed to examine the effect of probiotic bacteria Bacillus vireti 01 microencapsulated with G. folifera polysaccharide against freshwater prawn M. rosenbergii. Three different feeding trials were conducted for 15 days. The first group contained prawns fed with commercial diet. The second group was comprised of Aeromonas hydrophila challenged prawns fed with commercial feed. The third group consisted of A. hydrophila challenged prawns fed with microencapsulated probiotic-polysaccharide. Survival percentage was significantly decreased in prawns of group2 as compared to that of group1 and group3 prawns (p < 0.0001). The immunological parameters and antioxidant activities (p < 0.001) were found to be increased in group three prawns which were fed with encapsulated probiotic-seaweed polysaccharide and challenged with A. hydrophila as compared to that of group1 and group2. Tissue necrosis, fused lamella, haemocyte infiltration and damage of hepatopancreas lumen and tubule were noted in group2 prawns. There was no histological changes were observed in group3 prawns in which the histological architecture was similar to the control group1. The results suggested that combination of encapsulated probiotic B. vireti 01 and seaweed polysaccharide as dietary feed showed an enhancement of immune response, antioxidant activity and disease resistant of M. rosenbergii against A. hydrophila.

The hot-water extract of leaves of noni, Morinda citrifolia, promotes the immunocompetence of giant freshwater prawn, Macrobrachium rosenbergii.

The hot-water Morinda citrifolia leaf extract (HMLE) was prepared for in vitro assessment on phenoloxidase (PO) activity, respiratory bursts (RBs), and phagocytic activity (PA). Furthermore, the HMLE was administrated in the diet at 0.6, 3, and 6 g (kg diet)-1 for Macrobrachium rosenbergii, and the potential effects on the immunocompetence of prawns were evaluated. PO activity, RBs, and PA in hemocytes incubated with the HMLE at 140, 20, 20, and 140 mg l-1 significantly increased. The immune parameters of the total hemocyte count (THC), differential hemocyte count (DHC), RBs, PO activity, superoxide dismutase (SOD) activity, PA, transglutaminase (TG) activity and hemolymph clotting time were evaluated before and after 1, 3, 5, 7, and 9 weeks of the feeding trial. During 9 weeks of the feeding trial, higher THCs, DHCs, RBs, PO, and TG as well as accelerated clotting times were observed in prawns fed HMLE-containing diets at 0.6 g kg-1. The mRNA expressions of prophenoloxidase, TG, crustin, and lysozyme of prawns fed HMLE-containing diets at 0.6 g kg-1 for 9 weeks of the feeding trial significantly increased. The susceptibility of prawns fed the HMLE at 0.6 g kg-1 to Lactococcus garvieae infection significantly decreased, and the relative survival percentage was 23.1%. We therefore found that HMLE administrated through the diet at 0.6 g kg-1 was capable of enhancing the immunity and resistance against L. garvieae in M. rosenbergii.

Dietary Supplementation of Magnesium Oxide (MgO) Nanoparticles for Better Survival and Growth of the Freshwater Prawn Macrobrachium rosenbergii Post-larvae.

This study was performed to assess the growth-promoting potential of dietary magnesium oxide nanoparticles (MgO NPs) in Macrobrachium rosenbergii post-larvae (PL). MgO NPs were supplemented at 0, 100, 200, 300, 400 and 500 mg kg-1 with the basal diet (containing 0.95 g Mg kg-1); the concentrations of Mg in MgO NP-supplemented diets were increased correspondingly (1.07, 1.15, 1.24, 1.37 and 1.46 g Mg kg-1 respectively). MgO NP-supplemented diets were fed to M. rosenbergii PL (initial weight 0.11 ± 0.04 g) for a period of 90 days. In the carcasses of experimental prawns, the content of Mg was found to be elevated significantly with respect to the individual diet (102.14, 183.29, 205.46, 221.03, 237.10 and 254.36 µg Mg g-1 respectively) when compared with that of the control. The contents of Cu, Zn, Fe, Ca, Na and K levels were also found to be elevated in the carcasses of experimental prawns. Significant (P < 0.05) improvements were observed in nutritional indices [survival rate (SR), weight gain (WG), specific growth rate (SGR), feed conversion ratio (FCR) and protein efficiency ratio (PER)], activities of digestive enzymes (protease, amylase and lipase), concentrations of basic biochemical constituents (total protein, amino acid, carbohydrate, lipid, profiles of amino acids and fatty acids) and population of haemocytes [total and differential (hyalinocytes, semigranulocytes and granulocytes)] in all the test PL. Maximum performance was recorded in 500 mg kg-1 MgO NP-supplemented-feed-fed PL. There were no significant elevations recorded in activities of antioxidant enzymes [superoxide dismutase (SOD) and catalase (CAT)], lipid peroxidation (LPO) and metabolic enzymes [glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT)] recorded in any of the MgO NP-supplemented-feed-fed PL when compared with the control. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed increases in the staining intensity of polypeptide bands resolved in 500 mg kg-1 MgO NP-supplemented-feed-fed PL when compared with the control. Based on the gradual improvement in attaining survival, growth, FCR, biochemical constituents and haemocyte population, this study recommends MgO NP supplementation of 500 mg kg-1 for sustainable maintenance of M. rosenbergii PL. As the studied highest concentration of MgO NPs showed the best performance, it is necessary to study with beyond 500 mg kg-1 of MgO NPs to optimize the actual concentration.

Dietary supplementation of green synthesized manganese-oxide nanoparticles and its effect on growth performance, muscle composition and digestive enzyme activities of the giant freshwater prawn Macrobrachium rosenbergii.

The green synthesized Mn3O4 nanoparticles (manganese-oxide nanoparticles) using Ananas comosus (L.) peel extract was characterized by various techniques. HR-SEM photograph showed that manganese-oxide nanoparticles (Mn-oxide NPs) were spherical in shape, with an average size of 40-50 nm. The Zeta potential revealed the surface charge of Mn-oxide NPs to be negative. Further, the Mn-oxide NPs were dietary supplemented for freshwater prawn Macrobrachium rosenbergii. The experimental basal diets were supplemented with Mn-oxide NPs at the rates of 0 (control), 3.0, 6.0, 9.0, 12, 15 and 18 mg/kg dry feed weight. The as-supplemented Mn-oxide NPs were fed in M. rosenbergii for a period of 90 days. The experimental study demonstrated that prawns fed with diet supplemented with 3-18 mg Mn-oxide NPs/kg shows enhanced (P<0.05) growth performance, including final weight and weight gain (WG). Significant differences (P<0.05) in feed conversion ratio (FCR) were observed in prawn fed with different diets. Additionally, prawns fed with 3.0-18 mg/kg Mn-oxide NPs supplemented diets achieved significant (P<0.05) improvement in growth performance, digestive enzyme activities and muscle biochemical compositions, while, the prawns fed with 16 mg/kg of Mn-oxide NPs showed enhanced performance. Prawns fed on diet supplemented with 16 mg/kg Mn-oxide NPs showed significantly (P<0.05) higher total protein level. The antioxidants enzymatic activity (SOD and CAT) metabolic enzymes status in muscle and hepatopancreas showed no significant (P>0.05) alterations in prawns fed with 3.0-18 mg/kg of Mn-oxide NPs supplemented diets. Consequently, the present work proposed that 16 mg/kg of Mn-oxide NPs could be supplemented for flexible enhanced survival, growth and production of M. rosenbergii. Therefore, the data of the present study recommend the addition of 16 mg/kg of Mn-oxide NPs diet to developed prawn growth and antioxidant defense system.

The effect of copper nanoparticles supplementation on freshwater prawn Macrobrachium rosenbergii post larvae.

The present study was performed to assess the effects of dietary supplementation of copper nanoparticles (Cu-NPs) on growth, biochemical constituents, digestive enzyme activities, antioxidant, metabolic enzyme levels, and non specific immune response of the freshwater prawn, Macrobrachium rosenbergii post larvae (PL). The Cu-NPs (200 nm) were synthesized by facile and environmental friendly hydrothermal method. Cu-NPs were supplemented at 0, 10, 20, 40, 60, and 80 mg kg(-1) with the basal diets. These Cu-NPs supplemented diets were fed to M. rosenbergii PL for 90 days. Results showed significant (P<0.05) improvements were observed in survival, growth, digestive enzyme activities, concentrations of biochemical constituents and total and differential haemocytes count of prawns fed with 20 mg Cu-NPs kg(-1) supplemented feed. Prawns fed with 40-80 mg Cu-NPs kg(-1) supplemented feed showed negative performance. Activity of antioxidants and metabolic enzymes in the muscle and hepatopancreas of prawns showed no significant alterations (P>0.05) prawns fed with up to 20 mg Cu-NPs kg(-1) supplemented feeds. Whereas, prawns fed with 40-80 mg Cu-NPs kg(-1) supplemented feed showed significant (P<0.05) elevations in antioxidant and metabolic enzymes activities. Hence, 40-80 mg Cu-NPs kg(-1) diets may have toxic effect to M. rosenbergii. Hence, present study suggests that 20 mg Cu-NPs kg(-1) can be supplemented for regulating better survival, growth and immune response of M. rosenbergii PL.

Expression of biotransformation and oxidative stress genes in the giant freshwater prawn Macrobrachium rosenbergii exposed to chlordecone.

Chlordecone is a persistent organochlorine pesticide widely used between 1972 and 1993 in the French West Indies to control the root borer in banana fields. Chlordecone use resulted in long-term pollution of soils, contamination of waters, of aquatic organisms, and of fields. Chlordecone is known to be neurotoxic, to increase prostate cancer, and to have negative effects on cognitive and motor development during infancy. In Guadeloupe, most of the freshwater species living in contaminated rivers exceed the French legal limit of 20 µg·kg(-1) wet weight. In the present study, we chose a transcriptomic approach to study the cellular effects of chlordecone in the giant freshwater prawn Macrobrachium rosenbergii, an important economical species in Guadeloupe. Quantitative PCR revealed an induction of genes involved in defense mechanism against oxidative stress (catalase and selenium-dependent glutathione peroxidase) in prawns exposed to low environmental concentrations of chlordecone after 12 and 24 h of exposure. In prawns reared in a contaminated farm, transcription of genes involved in the biotransformation process (cytochrome P450 and glutathione-S-transferase (GST)) were induced after 8 days of exposure. Our results provide information on the mechanims of defense induced by chlordecone in aquatic crustacean species. This gene expression study of selected genes should be further strengthened by proteomic analyses and enzymatic activity assays to confirm the response of these biomarkers of stress in crustaceans and to give new insights into the mechanism of toxicity by chlordecone.

The effect of astaxanthin on resistance of juvenile prawns Macrobrachium nipponense (Decapoda: Palaemonidae) to physical and chemical stress / Efecto de la astaxantina en la resistencia al estrés físico y químico de los langostinos jóvenes de Macrobrachium nipponense (Decapoda: Palaemonidae)

In recent years, the use of new scientific techniques has effectively improved aquaculture production processes. Astaxanthin has various properties in aquacultureand its antioxidant benefits have been closely related to stress resistance; besides, it is an essential factor for growth in many crustaceans and fish. The objective of this study was to evaluate the resistance of prawn (Macrobrachium nipponense) fed diets containing different amounts of astaxanthin (AX) to the shock and stress of differentphysicochemical environments. A 70-day trial was conducted to evaluate the effect of supplementation of a source of astaxanthin (Carophyll Pink, 10% astaxanthin, w/w, Hoffman-La Roche, Switzerland) at various levels in the diet of M. nipponense juveniles. Four dry diets were prepared: AX0 without astaxanthin, AX50 with 50mg/kg, AX100 with 100mg/kg, and AX150 with 150mg/kg astaxanthin. The feeding trial was conducted in a recirculation water system consisting of 12 fiberglass tanks (1 000L) used for holding prawns. Three replicate aquaria were initially stocked with 36org/m² per tank. During the trial, prawns were maintained on a 12:12-h light:dark photoperiod with an ordinary incandescent lamp, and the water quality parameters were maintained as follows: water temperature, 25-26°C; salinity, 1g/L; pH, 8.5-8.8; dissolved oxygen, 6.0-6.5mg/L; and ammonia-nitrogen, 0.05mg/L. Incorporation of AX, production output, and physiological condition were recorded after 10 weeks of feeding. At the end of the growing period, the prawns were exposed to thermal shock (0°C), ammonia (0.75mg/L), and reduced oxygen (0.5mg/L). The time to lethargyand the time to complete death of the prawns were recorded. The results showed that control prawns had the shortest time to lethargy and death compared with prawns subjected to the other treatments. The results of this study have shown that the amount of muscle tissue and gill carotenoids in prawn fed with an AX150 diet showed greater reduction than those exposed to other treatments. It is possible that higher levels of astaxanthin in the body under oxygen reduction stress can be beneficial forprawns. These results suggest that male prawns showed lethargy earlier than females, and the percentage of carotenoid reduction in muscle and gill tissues was higher inmales. Rev. Biol. Trop. 62 (4): 1331-1341. Epub 2014 December 01.

En años recientes, la utilización de nuevas técnicas científicas ha tenido un efecto importante en mejorar los procesos de producción en acuicultura. La astaxantina tiene varias propiedades en la acuicultura y sus propiedades antioxidantes se encuentran estrechamente relacionadas con la resistencia al estrés. La astaxantina en muchos crustáceos y peces es un factor esencial para el crecimiento. El objetivo de este estudio fue evaluar la resistencia del langostino (Macrobrachium nipponense) alimentado con dietas conteniendo diferentes cantidades de astaxantina (AX), bajo diferentes condiciones de estrés ambiental. Un ensayo de 70 días fue llevado a cabo para evaluar el efecto de la suplementación de fuentes de astaxantina (Carophyll Pink, 10 % astaxanthin) en varios niveles en la dieta de jóvenes de M. nipponense. Cuatro dietas fueron preparadas: AX0 sin astaxantina, AX50 con 50mg/kg, AX100 con 100mg/kg y AX150 con 150mg/kg de astaxantina. Los ensayos de alimentación fueron conducidos en un sistema de recirculación de agua consistente en 12 estanques de fibra de vidrio (1 000L). Tres replicas fueron sembradas con 36org/m2 por tanque. Durante el experimento los langostinos fueron mantenidos con un fotoperiodo de 12:12 luz:oscuridad con lámparas incandescentes y los parámetros de la calidad del agua fueron mantenidos a 25-26°C la temperatura, 1 g/L la salinidad, 8.5-8.8 el pH, 6.0-6.5 mg/L el oxígeno y 0.05mg/L el nitrógeno amoniacal. La incorporación de la astaxantina, producción y condiciones fisicoquímicas fueron registradas después de 10 semanas de alimentación. Al final del periodo de crecimiento, los langostinos fueron expuestos a un shock térmico (0°C), amonio (0.75mg/L) y reducción de oxígeno 0.5 mg/L. El tiempo de letargia y el tiempo de muerte fueron registrados. Se encontró que la dieta con la mayor concentración de astaxantina (150mg/kg) presentó el mayor tiempo de letargia y la mayor concentración en branquias y músculo en el langostino M. nipponense.

Effects of hot-water extract of banana (Musa acuminata) fruit's peel on the antibacterial activity, and anti-hypothermal stress, immune responses and disease resistance of the giant freshwater prawn, Macrobrachium rosenbegii.

The hot-extracts isolated from fruit's peel of banana, Musa acuminata, was evaluated on the antibacterial activity to pathogens from aquatic animals, and immunostimulating potential, disease resistance and anti-hypothermal stress in giant freshwater prawn, Macrobrachium rosenbergii through injection administration. The banana peel extract (BPE) showed good activity against 1 Gram-positive and 3 Gram-negative pathogens, including Lactococcus garvieae, Photobacteria damsella, Vibrio alginolyticus and Vibrio parahemolyticus especially in prawn pathogen of L. garvieae strain, which were carried out by a disk diffusion method. Prawn received BPE via injection administration at 1-6 µg (g prawn)(-1) significantly increased total haemocyte count (THC), hyaline cell (HC), granular cell (GC), phenoloxidase (PO) activity and phagocytic activity against L. garvieae from 3 to 6 days, and significantly increased clearance efficiency against L. garvieae and a significantly decreased coagulation time of prawn from 1 to 6 days. Prawn injected with BPE at 6.0 µg (g prawn)(-1) for 6 days showed significantly increased superoxide dismutase (SOD) activity, but significantly decreased respiratory bursts (RBs) of per haemocyte. Survival rates of M. rosenbergii injected with BPE at concentrations of 1, 3 and 6 µg (g prawn)(-1) were significantly higher than those injected with saline control after challenge with L. garvieae for 4-6 days, and the respective relative survival percentages of prawn were 28.6%, 38.1%, and 47.8%, respectively at 6 days. The sublethal time of prawns that had received saline and BPE at 1, 3 and 6 µg (g prawn)(-1) for 6 days and then were transferred from 28 °C to 14 °C were 69.4, 79.8, 83.6, and 90.2 h, respectively. It was concluded that the BPE can be used as the bacteriostat, and immunostimulant and physiological regulator for prawn through injection administration to enhance immunity, physiological responses, and resistance against L. garvieae.

Dietary supplementation of zinc nanoparticles and its influence on biology, physiology and immune responses of the freshwater prawn, Macrobrachium rosenbergii.

The present study was conducted to assess the influence of dietary zinc nanoparticles (size 50 nm) on the growth, biochemical constituents, enzymatic antioxidant levels and the nonspecific immune response of the freshwater prawn, Macrobrachium rosenbergii post larvae (PL). The concentrations of dietary supplement zinc nanoparticles (ZnNPs) were 0, 10, 20, 40, 60 and 80 mg kg(-1) with the basal diet, and the level of Zn in ZnNP-supplemented diets were 0.71, 10.61, 20.73, 40.73, 60.61 and 80.60 mg kg(-1), respectively. ZnNP-incorporated diets were fed to M. rosenbergii PL (initial body weight, 0.18 ± 0.02 g) in a triplicate experimental setup for a period of 90 days. ZnNP supplemented feed fed PL up to 60 mg kg(-1) showed significantly (P < 0.05) improved performance in survival, growth and activities of digestive enzymes (protease, amylase and lipase). The concentrations of biochemical constituents (total protein, total amino acid, total carbohydrate and total lipid), total haemocyte count and differential haemocyte count were elevated in 10-60 mg kg(-1) ZnNP supplemented feed fed PL. However, the PL fed with 80 mg ZnNPs kg(-1) showed negative results. Activities of enzymatic antioxidants [superoxide dismutase (SOD) and catalase (CAT)], metabolic enzymes [glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT)] and the process of lipid peroxidation (LPO) in the hepatopancreas and muscle showed no significant alterations in 10-60 mg kg(-1) ZnNP supplemented feed fed PL. Whereas, 80 mg ZnNPs kg(-1) supplemented feed fed PL showed significant elevations in SOD, CAT, LPO, GOT and GPT. Therefore, 80 mg ZnNPs kg(-1) was found to be toxic to M. rosenbergii PL. Thus, the study suggests that up to 60 mg ZnNPs kg(-1) can be supplemented for regulating survival, growth and immunity of M. rosenbergii.

The effect of astaxanthin on resistance of juvenile prawns Macrobrachium nipponense (Decapoda: Palaemonidae) to physical and chemical stress.

In recent years, the use of new scientific techniques has effectively improved aquaculture production processes. Astaxanthin has various properties in aquaculture and its antioxidant benefits have been closely related to stress resistance; besides, it is an essential factor for growth in many crustaceans and fish. The objective of this study was to evaluate the resistance of prawn (Macrobrachium nipponense) fed diets containing different amounts of astaxanthin (AX) to the shock and stress of different physicochemical environments. A 70-day trial was conducted to evaluate the effect of supplementation of a source of astaxanthin (Carophyll Pink, 10% astaxanthin, w/w, Hoffman-La Roche, Switzerland) at various levels in the diet of M. nipponense juveniles. Four dry diets were prepared: AX0 without astaxanthin, AX50 with 50 mg/kg, AX100 with 100 mg/kg, and AX150 with 150 mg/kg astaxanthin. The feeding trial was conducted in a recirculation water system consisting of 12 fiberglass tanks (1000L) used for holding prawns. Three replicate aquaria were initially stocked with 36 org/m2 per tank. During the trial, prawns were maintained on a 12:12-h light:dark photoperiod with an ordinary incandescent lamp, and the water quality parameters were maintained as follows: water temperature, 25-26°C; salinity, 1 g/L; pH, 8.5-8.8; dissolved oxygen, 6.0-6.5 mg/L; and ammonia-nitrogen, 0.05 mg/L. Incorporation of AX, production output, and physiological condition were recorded after 10 weeks of feeding. At the end of the growing period, the prawns were exposed to thermal shock (0°C), ammonia (0.75 mg/L), and reduced oxygen (0.5 mg/L). The time to lethargy and the time to complete death of the prawns were recorded. The results showed that control prawns had the shortest time to lethargy and death compared with prawns subjected to the other treatments. The results of this study have shown that the amount of muscle tissue and gill carotenoids in prawn fed with an AX150 diet showed greater reduction than those exposed to other treatments. It is possible that higher levels of astaxanthin in the body under oxygen reduction stress can be beneficial for prawns. These results suggest that male prawns showed lethargy earlier than females, and the percentage of carotenoid reduction in muscle and gill tissues was higher in males.

Effect of crude oil petroleum hydrocarbons on protein expression of the prawn Macrobrachium borellii.

Hydrocarbon pollution is a major environmental threat to ecosystems in marine and freshwater environments, but its toxicological effect on aquatic organisms remains little studied. A proteomic approach was used to analyze the effect of a freshwater oil spill on the prawn Macrobrachium borellii. To this aim, proteins were extracted from midgut gland (hepatopancreas) of male and female prawns exposed 7 days to a sublethal concentration (0.6 ppm) of water-soluble fraction of crude oil (WSF). Exposure to WSF induced responses at the protein expression level. Two-dimensional gel electrophoresis (2-DE) revealed 10 protein spots that were differentially expressed by WSF exposure. Seven proteins were identified using MS/MS and de novo sequencing. Nm23 oncoprotein, arginine methyltransferase, fatty aldehyde dehydrogenase and glutathione S-transferase were down-regulated, whereas two glyceraldehyde-3-phosphate dehydrogenase isoforms and a lipocalin-like crustacyanin (CTC) were up-regulated after WSF exposure. CTC mRNA levels were further analyzed by quantitative real-time PCR showing an increased expression after WSF exposure. The proteins identified are involved in carbohydrate and amino acid metabolism, detoxification, transport of hydrophobic molecules and cellular homeostasis among others. These results provide evidence for better understanding the toxic mechanisms of hydrocarbons. Moreover, some of these differentially expressed proteins would be employed as potential novel biomarkers.

Effects of dietary administration of water hyacinth (Eichhornia crassipes) extracts on the immune responses and disease resistance of giant freshwater prawn, Macrobrachium rosenbergii.

The hot-water extract of Eichhornia crassipes leaves (ECE) was produced and incorporated into the diet of the prawn, Macrobrachium rosenbergii, as an immunostimulant. Survival rates of prawn against Lactococcus garvieae, and its immune parameters including the total haemocyte count (THC), different haemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, glutathione peroxidase (GPx) activity, transglutaminase (TG) activity, haemolymph coagulation time, and phagocytic activity and clearance efficiency against L. garvieae by M. rosenbergii were determined when prawn (23.0 ± 2.8 g) were fed ECE-containing diets at 0, 1.0, 2.0 and 3.0 g kg(-1). Prawn fed a diet containing ECE at 2.0 and 3.0 g kg(-1) for 12 days showed significantly increased THC, HC, GC, PO activity, RBs, SOD activity, GPx activity, and TG activity, and a significantly decreased coagulation time. The phagocytic activity and clearance efficiency against L. garvieae of prawn fed the ECE-containing diets at 1.0, 2.0, and 3.0 g kg(-1) were significantly higher than those of prawn fed the control diet at 3-12 days. Survival rates of M. rosenbergii fed the diet containing ECE at concentrations of 2 and 3 g kg(-1) were significantly higher than those fed the control diet after challenge with L. garvieae for 48-144 h. The relative percentage survival of prawn fed the 1.0, 2.0, and 3.0 g kg(-1) ECE-containing diets for 12 days were 17.5%, 39.1%, and 52.2%. It was concluded that the ECE can be used as an immunostimulant for prawn through dietary administration to enhance immune responses and resistance of M. rosenbergii against L. garvieae.

Retardation of post-mortem changes of freshwater prawn (Macrobrachium rosenbergii) stored in ice by legume seed extracts.

Meat quality of freshwater prawn (Macrobrachium rosenbergii) treated with soybean and bambara groundnut extracts at different concentrations was monitored during 10 days of iced storage. During storage, the control sample (without treatment) had a higher pH, TCA-soluble peptide content, heat soluble collagen content, proteolytic activities and psychrophilic bacterial count than did samples treated with soybean and bambara groundnut extracts. Conversely, shear force value and likeness scores of the control sample decreased (p<0.05), more likely associated with softening of muscle. The decrease in myosin heavy chain in the control sample was found after 6 days of storage. However, no changes in protein patterns of samples treated with soybean extracts at 2.5 mg/mL were found after 10 days of storage. Therefore, the injections of legume seed extracts, especially soybean extract, at a sufficient concentration, could be a means to retard muscle softening and maintain the qualities of freshwater prawn during iced storage.

Vitellogenin and lipovitellin from the prawn Macrobrachium borellii as hydrocarbon pollution biomarker.

During reproduction vitellogenin (VTG) is transported to vitellogenic oocytes as a precursor of egg yolk lipovitellin (LV). As VTG synthesis is affected by environmental stressors, it is widely used as biomarker in endocrine disruption studies. However, it has seldom been employed to evaluate invertebrate hydrocarbon pollution. An ELISA with anti-LV antibody was developed to evaluate the impact of water-soluble fraction of crude oil (WSF) on Macrobrachium borellii vitellogenesis. Prawn VTG concentration was within the range reported for other crustaceans; LV values were positively correlated with gonadosomatic index (GSI). Females at different vitellogenic stages were exposed to a sub-lethal concentration of WSF for 7 days. Exposed animals with GSI>7 increased their VTG and LV titer as compared to control organisms (190% and 140%, respectively). VTG levels in M. borellii were upregulated and highly sensitive to WSF exposure. This assay could be employed as a biomarker for freshwater hydrocarbon pollution.

Toxicity of crude oil to fresh water shrimp, Macrobrachium macrobrachion and Macrobrachium vollenhovenii, from Nigerian coastal water.

The water soluble fraction (WSF) of crude oil was tested against Macrobrachium macrobrachion and Macrobrachium vollenhovenii, at 2, 4, 6, 8 and 10 mg/L in glass aquaria stocked with ten animals for 96 h under observations for changes. Moribund swimming, restlessness, respiratory difficulties, depigmentation and mortalities were observed in the WSF exposure groups, but not in the controls. LC(50) values were estimated at 5 ± 1.76 and 4 ± 1.76 mg/L for M. macrobrachion and M. vollenhovenii respectively. There was no significant difference in mortalities between the two species (p > 0.05), leading to the conclusion that the WSF of crude oil in the Nigerian coastal waters may be equally toxic to M. macrobrachion and M. vollenhovenii.