Dietary administration of mangosteen, Garcinia mangostana, peel extract enhances the growth, and physiological and immunoendocrinological regulation of prawn, Macrobrachiumrosenbergii.

In this study, we investigated the potential immunostimulatory effects of mangosteen (Garcinia mangostana) peel extract on Macrobrachium rosenbergii, specifically in enhancing immunity and resistance against Lactococcus garvieae. We employed a dietary administration approach to assess the impact of different extract preparations from mangosteen peel, namely mangosteen peel powder (MPP), boiled mangosteen peel powder (MPB), and mangosteen peel extract (MPE). Following the administration of mangosteen peel extract, we evaluated growth performance, innate immune parameters, and disease resistance in the prawns. The results revealed a significant increase in total haemocyte count (THC), differential haemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (RBs), as well as phagocytic activity and clearance efficiency against L. garvieae. Based on these findings, we suggest that mangosteen peel extract can be utilized as an immunostimulant for prawns through dietary administration, regulating immune responses and enhancing resistance against pathogens by modulating carbohydrate metabolism.

17ß-Estradiol Induced Sex Reversal and Gonadal Transcriptome Analysis in the Oriental River Prawn (Macrobrachium nipponense): Mechanisms, Pathways, and Potential Harm.

Sex reversal induced by 17ß-estradiol (E2) has shown the potential possibility for monoculture technology development. The present study aimed to determine whether dietary supplementation with different concentrations of E2 could induce sex reversal in M. nipponense, and select the sex-related genes by performing the gonadal transcriptome analysis of normal male (M), normal female (FM), sex-reversed male prawns (RM), and unreversed male prawns (NRM). Histology, transcriptome analysis, and qPCR were performed to compare differences in gonad development, key metabolic pathways, and genes. Compared with the control, after 40 days, feeding E2 with 200 mg/kg at PL25 (PL: post-larvae developmental stage) resulted in the highest sex ratio (female: male) of 2.22:1. Histological observations demonstrated the co-existence of testis and ovaries in the same prawn. Male prawns from the NRM group exhibited slower testis development without mature sperm. RNA sequencing revealed 3702 differentially expressed genes (DEGs) between M vs. FM, 3111 between M vs. RM, and 4978 between FM vs. NRM. Retinol metabolism and nucleotide excision repair pathways were identified as the key pathways for sex reversal and sperm maturation, respectively. Sperm gelatinase (SG) was not screened in M vs. NRM, corroborating the results of the slice D. In M vs. RM, reproduction-related genes such as cathepsin C (CatC), heat shock protein cognate (HSP), double-sex (Dsx), and gonadotropin-releasing hormone receptor (GnRH) were expressed differently from the other two groups, indicating that these are involved in the process of sex reversal. Exogenous E2 can induce sex reversal, providing valuable evidence for the establishment of monoculture in this species.

Effects of dietary soluble ß-1,3-glucan on the growth performance, antioxidant status, and immune response of the river prawn (Macrobrachium nipponense).

The effects of dietary ß-1,3-glucan on the growth performance, body composition, hepatopancreas tissue structure, antioxidant activities, and immune response of the river prawn (Macrobrachium nipponense) were investigated. In total, 900 juvenile prawns were fed one of five diets with different contents of ß-1,3-glucan (0%, 0.1%, 0.2%, and 1.0%) or 0.2% curdlan for 6 weeks. The growth rate, weight gain rate, specific growth rate, specific weight gain rate, condition factor, and hepatosomatic index of juvenile prawns fed 0.2% ß-1,3-glucan were significantly higher than those fed 0% ß-1,3-glucan and 0.2% curdlan (p < 0.05). The whole-body crude lipid content of prawns supplemented with curdlan and ß-1,3-glucan was significantly higher than that of the control group (p < 0.05). The antioxidant and immune enzyme activities of superoxide dismutase (SOD), total antioxidant capacity (T-AOC), catalase (CAT), lysozyme (LZM), phenoloxidase (PO), acid phosphatase (ACP), and alkaline phosphatase (AKP) in the hepatopancreas of juvenile prawns fed 0.2% ß-1,3-glucan were significantly higher than those of the control and 0.2% curdlan groups (p < 0.05), and tended to increase and then decrease with increasing dietary ß-1,3-glucan. The highest malondialdehyde (MDA) content was observed in juvenile prawns without ß-1,3-glucan supplementation. The results of real-time quantitative PCR indicated that dietary ß-1,3-glucan promoted expression of antioxidant and immune-related genes. Binomial fit analysis of weight gain rate and specific weight gain rate showed that the optimum ß-1,3-glucan requirement of juvenile prawns was 0.550%-0.553%. We found that suitable dietary ß-1,3-glucan improved juvenile prawns growth performance, antioxidant capacity, and non-specific immunity, which provide reference for shrimp healthy culture.

Scutellaria polysaccharide mediates the immunity and antioxidant capacity of giant freshwater prawn (Macrobrachium rosenbergii).

The giant freshwater prawn (Macrobrachium rosenbergii) is a commercially valuable freshwater crustacean species that frequently appears a death affected by various diseases, resulting in substantial economic losses. Improving the survival rate of M. rosenbergii is a hot and essential issue for feeding the prawns. Scutellaria polysaccharide (SPS) extracted from Scutellaria baicalensis (a Chinese medicinal herb) is conducive to the survival rate of organisms by enhancing immunity and antioxidant ability. In this study, M. rosenbergii was fed 50, 100, and 150 mg/kg of SPS. The immunity and antioxidant capacity of M. rosenbergii were tested by mRNA levels and enzyme activities of related genes. The mRNA expressions of NF-κB, Toll-R, and proPO (participating in the immune response) in the heart, muscle, and hepatopancreas were decreased after four weeks of SPS feeding (P < 0.05). This indicated that long-term feeding of SPS could regulate the immune responses of M. rosenbergii tissues. The activity levels of antioxidant biomarkers, alkaline phosphatase (AKP), and acid phosphatase (ACP) had significant increases in hemocytes (P < 0.05). Moreover, catalase (CAT) activities in the muscle and hepatopancreas, as well as superoxide dismutase (SOD) activities in all tissues, significantly decreased after four weeks of culture (P < 0.05). The results demonstrated that long-term feeding of SPS could improve the antioxidant capacity of M. rosenbergii. In summary, SPS was conducive to regulating the immune capacity and enhancing the antioxidant capacity of M. rosenbergii. These results provide a theoretical basis for supporting SPS addition to the feed of M. rosenbergii.

The immunostimulatory effects of Theobroma cacao L. pod husk extract via injection and dietary administrations on Macrobrachium rosenbergii and its resistance against Lactococcus garvieae.

The extract from Theobroma cacao L, pod husk served as the immunostimulant to enhance the immunity and resistance against Lactococcus garvieae of Macrobrachium rosenbergii. In this study, we employed the injection method and dietary administration method to determine the effect of cocoa pod husk (CPH) on M. rosenbergii. The non-specific immune parameters and disease resistance were evaluated after the prawn injected with 1 µg prawn-1 CPH extract (C1), 3 µg prawn-1 CPH extract (C3), and 5 µg prawn-1 CPH extract (C5) for 1, 3, and 7 days. The results showed a significant increase of total haemocyte count (THC), differential haemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (RBs), and phagocytic activity and clearance efficiency to L. garvieae. The non-specific immune parameters, physiological parameters, and disease resistance and growth performance were evaluated after the prawn fed with 1 g kg-1 CPH extract diet (CD1), 3 g kg-1 CPH extract diet (CD3) and 5 g kg-1 CPH extract diet (CD5). The results showed a significant increase in all immune parameters and showed a significant decrease in physiological parameters. No significant difference was observed in growth performance of prawn fed with the CPH containing diet. Both injection and dietary method showed a significant increase in disease resistance against to L. garvieae. We therefore recommend that CPH extract can be used as a immunostimulant for prawn by dietary administration to regulate immune responses, and carbohydrate metabolism lead to enhance resistance against pathogen.

Influence of Garlic (Allium sativum) Clove-Based Selenium Nanoparticles on Status of Nutritional, Biochemical, Enzymological, and Gene Expressions in the Freshwater Prawn Macrobrachium rosenbergii (De Man, 1879).

Selenium (Se) is one of the essential micronutrients for performing vital body functions. This study aims at examining the influence of dietary supplementation of garlic clove-based green-synthesized selenium nanoparticles (GBGS-SeNPs, 48-87 nm) on carcass minerals and trace elements, and growth, biochemical, enzymological, and gene expression analyses in the freshwater prawn, Macrobrachium rosenbergii post larvae (PL). The 96 h LC50 of this GBGS-SeNPs to M. rosenbergii PL was 52.23 mg L-1. Five different artificial diets without supplementation of GBGS-SeNPs (control, 0.0 mg kg-1) and with supplementations of GBGS-SeNPs starting from 100 times lower than the LC50 value (0.5, 1.0, 1.5, and 2.0 mg kg-1) were prepared and fed to M. rosenbergii PL for 90 days. A dose-dependent accumulation of Se was observed in the carcass of experimental prawns. GBGS-SeNPs, up to 1.5 mg kg-1 significantly influenced the absorption of other trace elements (Ca, Cu, and Fe) and mineral salts (K, Mg, Na, and Zn). GBGS-SeNPs-supplemented diets showed efficient food conversion ratio (FCR) of 1.32 g against 2.71 g, and therefore enhanced the survival rate (85.6% against 78.8% in control) and weight gain (WG) of 1.41 g against 0.46 g of control prawn. GBGS-SeNPs significantly elevated the activities of protease, amylase, and lipase, and the contents of total protein, essential amino acids (EAA), total carbohydrate, total lipid, monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), and ash. These indicate the growth promoting potential of GBGS-SeNPs in prawn. The insignificantly altered activities of glutamic oxaloacetate transaminase (GOT), glutamic pyruvate transaminase (GPT), superoxide dismutase (SOD), and catalase, and the content of malondialdehyde (MDA) up to 1.5 mg kg-1 suggest its acceptability in prawn. Moreover, a respective down- and upregulated myostatin (MSTN) and crustacean hyperglycemic hormone (CHH) genes confirmed the influence of GBGS-SeNPs on the growth of prawn. In contrast, 2.0 mg kg-1 GBGS-SeNPs supplementation starts to produce negative effects on prawn (FCR, 1.76 g; survival rate, 82.2%; WG, 0.84 g against respective values of 1.32 g, 85.6%; and 1.41 g observed in 1.5 mg kg-1 of GBGS-SeNPs-supplemented diet fed prawn). This study recommends a maximum of 1.5 mg kg-1 GBGS-SeNPs as dietary supplement to attain sustainable growth of M. rosenbergii. This was confirmed through polynomial and linear regression analyses.

Low fish meal diet supplemented with probiotics ameliorates intestinal barrier and immunological function of Macrobrachium rosenbergii via the targeted modulation of gut microbes and derived secondary metabolites.

The unsuitable substitution ratio of fish meal by plant protein will reshape the intestinal microbial composition and intestine immunity. However, previous studies were mostly limited to investigating how different feed or probiotics characterized the microbial composition but ignored the biological interactions between bacteria and host physiology through secondary metabolites. Therefore, this study integrates the apparent indicators monitoring, 16S rDNA sequencing, and metabonomics to systematically investigate the effects of cottonseed protein concentrate (CPC) substitution of fish meal and Bacillus coagulans intervention on gut microbes, secondary metabolites, and intestinal immunity of Macrobrachium rosenbergii. Prawns were fed with three diets for 70 days: HF diets contained 25% fish meal, CPC in LF diets were replaced with 10% fish meal, and LF diets supplemented with 2 × 108 CFU/g diet B. coagulans were designated as BC diets. Results showed that CPC substitution induced a significant decrease in digestive enzyme activities (trypsin and lipase) and gut barrier protein PT-1 expression and a significant increase in γ-GT enzyme activity and inflammatory-related factors (Relish and Toll) expression. B. coagulans treatment mitigated the negative changes of the above indicators. Meanwhile, it significantly improved the expression levels of the barrier factor PT-1, the reparative cytokine IL-22, and Cu/Zn-SOD. CPC substitution resulted in a remarkable downregulated abundance of Firmicutes phyla, Flavobacterium spp., and Bacillus spp. B. coagulans treatment induced the callback of Firmicutes abundance and improved the relative abundance of Sphingomonas, Bacillus, and Ralstonia. Functional prediction indicated that CPC substitution resulted in elevated potential pathogenicity of microbial flora, and B. coagulans reduces the pathogenesis risk. Pearson's correlation analysis established a significant positive correlation between differential genera (Sphingomonas, Bacillus, and Ralstonia) and secondary metabolites (including sphingosine, dehydrophytosphingosine, amino acid metabolites, etc.). Meanwhile, the latter were significantly associated with intestinal immunoregulation-related genes (Cu/Zn-SOD, IL-22, PT-1, Toll, and Relish). This study indicated that B. coagulans could mediate specific gut microbes and the combined action of multiple functional secondary metabolites to affect intestinal barrier function, digestion, and inflammation. Our study revealed the decisive role of gut microbes and derived secondary metabolites in the model of dietary composition-induced intestinal injury and probiotic treatment from a new perspective.

Molecular isolation and expression analysis of hemocyanin isoform 2 of Macrobrachium rosenbergii.

OBJECTIVE: Hemocyanin is a copper-bearing protein in the hemolymph of many arthropods and mollusks and functions as an oxygen transport and important nonspecific immune protein. METHODS: In this study, complementary DNA of hemocyanin isoform 2 of the prawn Macrobrachium rosenbergii (MrHc2) was isolated by rapid amplification of cDNA ends and mRNA expression was characterized to elucidate molecular basis of its function. RESULT: With a molecular mass of 77.3 kDa, MrHc2 contained three domains: hemocyanin-all-alpha, hemocyanin-copper-containing, and hemocyanin-immunoglobulin-like domains. Molecular phylogenetic analysis revealed that MrHc2 belongs to the γ-type subunit and is closely related to hemocyanin subunit 1 of the palaemonid shrimp Macrobrachium nipponense. In addition, MrHc2 resided in a different clade relative to hemocyanin (MrHc) of M. rosenbergii (α-type subunit) and in a different subclade relative to the hemocyanin proteins of penaeid shrimp. The messenger RNA transcript of MrHc2 was highly expressed in the hepatopancreas and weakly expressed in the gills, intestine, stomach, muscle, and hemocytes. Upon challenge with M. rosenbergii nodavirus (MrNV), the expression of MrHc2 was 1.96-, 2.93-, and 1.96-fold on days 3, 4, and 5, respectively, and then gradually declined to basal levels on day 7. CONCLUSION: This study suggests that MrHc2 plays an important role in the innate immune response of M. rosenbergii to MrNV.

Effects of rice-prawn (Macrobrachium nipponense) co-culture on the microbial community of soil.

In the Lixiahe region of China, co-culture has been rapidly promoted in flooded paddy fields owing to its ecological and economic benefits. Rice-prawn co-culture can reduce the damage of crab and shrimp to rice growth and paddy field and substantially change the soil microbial community and soil fertility. In this study, we compared changes in the soil microbial community and soil fertility in waterlogged paddies under conventional rice culture (CR), rice-prawn (Macrobrachium nipponense) co-culture (RP), and pond culture (PC). The microbial abundance in RP was significantly higher than that in CR. RP soil microbial diversity was significantly higher than PC soil microbial diversity. The dominant bacteria in RP soil were Proteobacteria, Chloroflexi, and Bacteroidetes. Compared with those in CR, total organic matter (TOM) and total nitrogen in RP were relatively stable, available potassium and available phosphorus (AP) decreased, and other indicators increased significantly. Soil fertility significantly benefited from co-culture, with total organic carbon (TOC) increasing. Interactive relationship analysis showed that TOM, TOC, AP, and NH4+-N were the main factors affecting the microbial community. Co-occurrence network analyses showed that network modularity increased with co-culture, indicating that a unique soil microbial community formed under co-culture, improving the adaptability and tolerance to co-culture. Thus, RP is a suitable culture method for this commercially important species. The results of this study can inform the practical operation of fertilizer use and sustainable development of rice-prawn aquaculture systems. KEY POINTS: • Microbial abundance and diversity increased under rice-prawn co-culture. • Co-culture significantly improved soil fertility, with an increase in TOC. • Rice-prawn co-culture is an ecologically suitable culture method for prawns.

Dietary thiamine modulates carbohydrate metabolism, antioxidant status, and alleviates hypoxia stress in oriental river prawn Macrobrachium nipponense (de Haan).

Hypoxia is one of the challenges in prawns aquaculture. However, the role of thiamine, which is a coenzyme in carbohydrate metabolism with antioxidant properties, in reducing hypoxia in prawns aquaculture is currently unknown. We investigated the effects of thiamine on antioxidant status, carbohydrate metabolism and acute hypoxia in oriental river prawn, Macrobrachium nipponense. One thousand eight hundred prawns (0.123 ± 0.003 g) were fed five diets (60 prawns each tank, six replicates per diet) supplemented with graded thiamine levels (5.69, 70.70, 133.67, 268.33 and 532.00 mg/kg dry mater) for eight weeks and then exposed to hypoxia stress for 12 h followed by reoxyegnation for 12 h. The results showed that, under normoxia, prawns fed the 133.67 or 268.33 mg/kg thiamine diet had significantly lower glucose 6-phosphatedehydrogenase, succinate dehydrogenase and phosphoenolpyruvate carboxykinase activities than those fed the other diets. Moreover, total antioxidant capacity (T-AOC) increased significantly when prawns were fed the 133.67 mg/kg thiamine diet. Superoxide dismutase (SOD) activity and malonaldehyde (MDA) content also increased significantly when prawns were fed the 268.33 or 532.00 mg/kg thiamine diet under hypoxia. And the significantly increased SOD activity and MDA level also observed in prawns fed 532.00 mg/kg thiamine under reoxygenation. Under normoxia, prawns fed the 70.70 or 133.67 mg/kg thiamine diet decreased the mRNA expressions of AMP-activated protein kinase-alpha (AMPK-α), pyruvate dehydrogenase-E1-α subunit (PDH-E1-α) and hypoxia-inducible factor-1s (HIF-1α, HIF-1ß), but increased the mRNA expressions of phosphofructokinase (PFK) significantly. After 12 h of hypoxia, the energy metabolism related genes (AMPK-ß, AMPK-γ, PFK, PDH-E1-α), hypoxia-inducible factor related genes (HIF-1α, HIF-1ß) and thiamine transporter gene (SLC19A2) were up-regulated significantly in prawns fed the 133.67 or 268.33 mg/kg thiamine diets. After 12 h of reoxygenation, prawns fed the 133.67 or 268.33 mg/kg diet significantly decreased the SOD activity, MDA level and SLC19A2 mRNA expression compared with other diets. The optimum thiamine was 161.20 mg/kg for minimum MDA content and 143.17 mg/kg for maximum T-AOC activity based on cubic regression analysis. In summary, supplementing 143.17 to 161.20 mg/kg thiamine in the diets for M. nipponense improves the antioxidant capacity under normoxia and reduces the oxidative damage under hypoxia stress.

Effects of high fat in the diet on growth, antioxidant, immunity and fat deposition of Macrobrachium rosenbergii post-larvae.

Lipids are essential nutrients for organisms, and high-fat feeds for shrimp may cause oxidative stress. This study evaluated the effects of feeding high fat in the diet on the growth, antioxidant, immunity, and liver fat accumulation of Macrobrachium rosenbergii post-larvae. Five groups with an initial body weight of 0.0084 ± 0.001 g were fed five isonitrogenous and isoenergetic diets (47.01% crude protein and 18.40 kJ/g gross energy) containing 8%, 10%, 12%, 14% and 16% (named L8, L10, L12, L14 and L16) lipid for 8 weeks, respectively. The results showed that the weight gain rate (WGR) and specific growth rate (SGR) of L8 group were significantly higher than those of L10, L12, L14 and L16 group (P < 0.05), and the feed coefficient (FCR) of L8 group was significantly lower than that of other groups (P < 0.05). With the increase of dietary fat level, the content of MDA and the activity of SOD increased significantly, and the activities of T-AOC and CAT decreased significantly (P < 0.05). H&E staining clearly revealed the occurrence of hepatocyte swelling, hepatocyte vacuolization and nucleus displacement to the peripheral cell vacuolization in the L16 group, and hepatic lipid accumulation was further observed in the L14 and L16 group by Oil red O staining. In addition, high-fat diet significantly upregulated the expression of Dorsal, Relish and IκBα mRNA, and also upregulated the expression of fat synthesis-related genes FAS, ACC, DGAT and fat transport-related gene FABP (P < 0.05), and significantly downregulated the expression of fat metabolism-related genes AMPK and CPT-1 (P < 0.05) compared to that of the L8 group. In conclusion, this study showed that feeding a high-fat diet could induce oxidative stress, inhibit growth performance, alter antioxidant capacity, cause hepatic fat deposition and affect the immune system of M. rosenbergii post-larvae.

Comparative Proteomic Analysis Revealed the Mechanism of Tea Tree Oil Targeting Lipid Metabolism and Antioxidant System to Protect Hepatopancreatic Health in Macrobrachium rosenbergii.

Tea tree oil (TTO) is a pure natural plant essential oil. The studies evaluated the hepatopancreas lipid metabolism and antioxidant efficacy of Macrobrachium rosenbergii fed with 0 (CT group) and 100 mg/kg TTO (TT group) by label-free quantification proteomic analysis. Compared to the CT group, the TT group improved growth performance and increased the survival rate after stress. Dietary TTO also decreased hemolymph AST and ALT activities and decreased hepatopancreatic vacuolation. At the same time, hepatopancreas lipids droplets and hemolymph lipids (TG, TC, LDL-C) were decreased, and the peroxidation products content (MDA, LPO, 4-HNE) was also decreased. In addition, the levels of hepatopancreas antioxidant enzymes (T-AOC, CAT, and SOD) were increased in the TT group. With proteomic analysis, a total of 151 differentially expressed proteins (DEPs) (99 up-regulated and 52 down-regulated) were identified in the hepatopancreas. Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein interaction analysis showed that the 16 DEPs have interactions, which are mainly involved in the pathways related to lipid metabolism (fatty acid biosynthesis, fatty acid metabolism, glycerophospholipid metabolism) and redox reaction (cytochrome P450 enzyme systems). Furthermore, the mRNA expression of 15 proteins followed the proteomic analysis with qRT-PCR validation. Pearson correlation analysis showed that fatty acids and glycerophospholipid metabolism-related proteins were highly correlated to peroxide content, glycerophospholipid metabolism, and cytochrome P450 system-related proteins (CYP1A1, GSTT1, GPX4) were highly correlated to AST and ALT. Additionally, GPX4 is closely related to peroxide content and antioxidant enzyme activity. Our results revealed that TTO plays a protective role in the hepatopancreas targeting the critical enzymes and antioxidant reactions in lipid metabolism. Provides a new perspective to elucidate the action path of TTO in protecting invertebrate hepatopancreas, highlights the influence of lipid metabolism on hepatopancreas health and the interaction between lipid metabolism and antioxidant system in the regulation of TTO.

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 garlic (Allium sativum) extract on growth, enzymological and biochemical responses and immune-related gene expressions in giant freshwater prawn (Macrobrachium rosenbergii).

In the current study, growth performance, biochemical constituents of muscle, activities of enzymes in the haemolymph, and expressions of immune-related genes were evaluated in the giant freshwater prawns Macrobrachium rosenbergii fed diets supplemented with aqueous garlic (Allium sativum) extract at 0, 5, 10 and 20 g/kg (w/w) for 60 days. At the end of the feeding trial, weight gain and specific growth rate were significantly improved in garlic-fed prawn groups compared with the control (p < 0.05). Moreover, feed conversion ratio was significantly lower in the garlic-fed groups than in the control (p < 0.05). Activities of catalase, superoxide dismutase and glutathione peroxidase in the hepatopancreas, activities of alanine aminotransferase, aspartate aminotransferase and levels of albumin and total protein in the hemolymph were significantly increased in the garlic treatments (p < 0.05). Furthermore, garlic supplemented diets improved muscle biochemical profiles, particularly contents of crude protein and total ash and upregulations of immune deficiency and heat shock proteins (HSP70) gene expression (p < 0.05). Therefore, garlic has positive effects on growth performance and physio-biochemical responses of M. rosenbergii, and thus, it can be used as an additive for stress resistance and as a growth promoter in sustainable aquaculture.

Effects of dietary tea tree oil on the growth, physiological and non-specific immunity response in the giant freshwater prawn (Macrobrachium rosenbergii) under high ammonia stress.

This study aimed to investigate the effects of dietary tea tree oil (TTO) on the performance, intestinal antioxidant capacity, and non-specific immunity after ammonia nitrogen stress in Macrobrachium rosenbergii. Six experimental diets were formulated with 0, 25, 50, 100, 200, 400 mg/kg TTO, respectively. A total of 900 prawns (average initial weight, 0.39 ± 0.01 g) were randomly assigned to 6 groups in triplicate in 18 tanks. After an 8-week feeding trial, 20 prawns from each tank were changed with 20 mg/L ammonia stress for 24 h. The results showed that 100 mg/kg TTO significantly increased prawns performance and survival rate compared with the control group. Moreover, 100 and 200 mg/kg TTO significantly improved intestinal antioxidant capabilities by increasing SOD enzyme activities and decreasing MDA levels. In addition, the prawns fed with 100 mg/kg TTO diet showed the highest survival rate under ammonia stress. After ammonia stress, the group of 100 mg/kg TTO significantly improved antioxidant capacity by increasing hemolymph respiratory burst activity, as well as intestinal anti-superoxide anion activity and SOD. Coincidentally, 100 mg/kg TTO significantly upregulated the intestinal relative expression of antioxidant-related genes (peroxiredoxin-5). Further, it was found that 100 mg/kg TTO activated the toll-dorsal pathway in prawns, which performed the similar function as the classic NF-κB pathway by upregulating the TNF-α and IL-1. Finally, 100 mg/kg TTO increased the levels of iNOS activities and NO contents after ammonia stress and enhanced non-specific immunity. The results indicated that 100 mg/kg TTO could significantly improve the M. rosenbergii performance, antioxidant capacity and ammonia stress resistance. We suggested that the mechanisms may be attributed to that TTO enhanced the antioxidant capacity and non-specific immunity of M. rosenbergii via the NF-κB signal pathway.

Impact of different dietary lutein levels on growth performance, biochemical and immuno-physiological parameters of oriental river prawn (Macrobrachium nipponense).

A 56-day trial was conducted to evaluate the effects of dietary lutein pigment on growth, biochemical, and immuno-physiological parameters of the oriental river prawn. Prawns were fed five formulated diets containing different lutein levels, 0 (control), 50, 100, 150, and 200 mg/kg. Growth performance, except hepatosomatic index, was affected by different lutein levels, and biochemical parameters (urea, uric acid, glucose, creatinine, and triglycerides) decreased. However, high-density and low-density lipoprotein elevated significantly compared to the control treatment. Furthermore, calcium, phosphorus, and cholesterol did not show a significant difference. Hemato-immunological parameters (albumin, total protein, cortisol, lysozyme, phenoloxidase, total hemocyte count, granular cells, semi-granular cells, hyaline cells, alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase), and hepatopancreatic antioxidant statuses (total antioxidant capacity, superoxide dismutase, catalase, and malondialdehyde), were significantly affected; however, alkaline phosphatase and glutathione peroxidase were not affected by lutein treatments. By increasing dietary lutein levels, digestive enzyme activities, total bacteria count, total carotenoid content, significantly increased. Conversely, lactic acid bacteria were not affected. Overall, the research results demonstrated that adding 200 mg/kg of lutein to the diet improved growth performance, biochemical and immuno-physiological parameters of the oriental river prawn.

Opaepupu, a new genus and species of bivalve-associated shrimp (Decapoda: Caridea: Palaemonidae) from Hawai'i.

A new palaemonid shrimp genus, Opaepupu gen. nov., is established to accommodate a new species of bivalve-associated shrimp, Opaepupu huna sp. nov. from Hawaii. A single mated pair, the female holotype and the male allotype, were found inside the trapezid bivalve Trapezium oblongum (Linnaeus, 1758) at a depth of 14 m in Kâne'ohe Bay, Hawai'i. The new genus is characterised by the rostrum being proximally broad, distally pointed, mid-dorsally carinate, and non-dentate; the anterolateral margin of the carapace without supraorbital, hepatic or epigastric teeth, but with a strong sharp antennal tooth; the sixth pleonite posteriorly unarmed; the telson medially depressed, with the dorsal surface armed with two pairs of submarginal cuspidate setae and with the posterior margin armed with two pairs of spiniform setae; the distolateral angle of the first article of the antennular peduncle without a sharp tooth; the mandible without a palp; the maxillular palp furnished with one long stiff seta dorsal to a small tooth-like extension; the first maxilliped without a palp; the third maxilliped not being operculate; the second pereiopods moderately robust, relatively slender, subequal, subsymmetrical, with simple teeth on the cutting edges of the fingers; the ambulatory pereiopods being slender, each ending in an elongate biunguiculate dactylus; and the uropodal exopod with a faint diaeresis and greatly reduced distolateral spiniform seta. The phylogenetic position of Opaepupu gen. nov. remains unclear, although it does not appear to be closely related to other bivalve-associated palaemonid genera.

N-acetyl-ß-d-glucosaminidase activity in Palaemon serratus - Methodological optimisation and intrinsic variability.

Chitinolytic enzymes fulfil a key role in the moulting process of crustaceans, in degrading the endocuticle during apolysis. Measuring the enzyme activity is an interesting manner to monitor the moult process at sub-individual level, complementary to the classical observation of the integument morphogenesis, ecdysis success, or moult cycle duration. The present study aimed to optimise the methodology of using N-acetyl-ß-D-glucosaminidase (NAGase) activity to monitor moulting in the marine prawn Palaemon serratus, and to compare NAGase activity levels along the moult cycle of both male and female specimens. First, to optimise protocols for five different organs, different reaction medium compositions were tested, considering the type buffer, concentration of the substrate, and the load in enzymatic extract. Second, levels of NAGase activity were closely monitored during eight moulting stages in male prawns. Variations in NAGase activity were observed during the moult cycle, with an increase in activity in the late premoult phase of approximately 2.4-fold the level of the intermoult phase. This response profile was observed for each tested organ. The levels of NAGase activity of male and female specimens were compared during three stages of the premoult phase. The patterns observed for both sexes were similar for all the tested organs.

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.

Effects of graded levels of mineral mixtures (Aquamin® and Agrimin®) supplemented diets on growth, survival, proximate composition, and carcass mineralization of juvenile freshwater prawn, Macrobrachium malcolmsonii (H Milne-Edwards 1844).

The present study investigated the effects of Aquamin® mineral mixture (AQMM) and Agrimin® mineral mixture (AGMM) supplemented diets on growth and chemical composition of juvenile freshwater prawn Macrobrachium malcolmsonii (H. Milne-Edwards, 1844). Experimental diets containing 6 different AQMM levels (Trial 1: 0, 0.5, 1.0, 1.5, 2.0, and 2.5%) and 6 different AGMM levels (Trial 2: 0, 0.5, 1.0, 1.5, 2.0, and 2.5%) were formulated to feed juvenile prawn [initial body weight of 0.82 ± 0.05 g (Trial 1) and 1.24 ± 0.03 g (Trial 2)] for 60 days. Prawn fed diets containing 1.0% of AQMM and AGMM showed significantly increased (P < 0.05) survival rate, weight gain, specific growth rate, protein efficiency ratio, edible flesh weight, and proximate composition (moisture, crude protein, crude fat, and ash), while feed intake and feed conversion ratio were significantly decreased (P < 0.05). Whereas, 1.5-2.5% of AQMM- and AGMM-supplemented diets fed prawn showed a reverse trend when comparing other groups. Moreover, the minerals (macro elements: Ca, P, Mg, Na, and K; trace elements: Cu, Zn, and Fe) were significantly increased (P < 0.05) in the carcass of prawn when fed with 2.5% AQMM- and AGMM-supplemented diets. The present results suggest that the optimal dietary supplementation of AQMM and AGMM at a concentration of up to 1.06 and 1.02%, respectively (based on polynomial regression analysis), improved growth and enhanced the crude protein level of juvenile prawn.