Influence of Different Feed Particle Sizes on the Growth Performance and Nutrition Composition in Crayfish, Procambarus clarkii Larvae.

A suitable feed size has a positive effect on animal feeding. For aquatic larvae, the correct feed size is very important for their growth. This experiment analyzed and compared the effect of different particle sizes of feed for larval stages on the growth performance, whole body composition, and muscle amino acid and fatty acid composition of crayfish. Five larval crayfish diets of different particle sizes, namely < 0.40 mm (Group A, control group), 0.40-0.50 mm (Group B), 0.71-0.85 mm (Group C), 0.90-1.00 mm (Group D) and 1.5 mm (Group E), were fed to 2000 crayfish (initial weight 0.0786 ± 0.0031 g) for 100 d. The results showed that as the particle size increased, final weight, weight gain (WG, p = 0.001) and specific growth rate (SGR, p = 0.000) of the crayfish tended to increase and then leveled off, with the control group being the lowest. The feed conversion ratio (FCR, p = 0.000) showed a decreasing and then equalizing trend with increasing particle size, but there was no significant difference between the groups except the control group. Broken-line regression analysis showed that the critical values for the appropriate particle feed size for crayfish larvae were 0.55 mm and 0.537 mm using SGR and FCR as indicators. Groups B, C and D had the highest crude protein content and were significantly higher than the control group (p = 0.001). Group E had the highest umami amino acid (UAA) and was significantly higher than the control group (p = 0.026). The content of isoleucine (Ile, p = 0.038) and phenylalanine (Phe, p = 0.038) was highest in group C and significantly higher than in the control group. Through principal component analysis, groups C and D were shown to contain leucine (Leu), glutamic (Glu), methionine (Met), valine (Val), histidine (His), Phe, and Ile levels significantly induced. The content of linoleic acid (C18:2n6, p = 0.000), linolenic acid (C18:3n3, p = 0.000), saturated fatty acid (SFA, p = 0.000), monounsaturated fatty acid (MUFA, p = 0.001), polyunsaturated fatty acid (PUFA, p = 0.000) and n-6 PUFA (p = 0.000) in group C was the highest and significantly higher than the control group. Principal component analysis showed that group C significantly induced the levels of C18:2n6, C18:3n3, DHA, EPA, n-3 PUFA and n-6 PUFA in muscle. Therefore, our results suggest that appropriate feed particle size can improve the growth performance and nutrient composition of crayfish. Based on the broken-line regression analysis of SGR and FCR, the critical values of optimal particle size for crayfish are 0.55 mm and 0.537 mm, and when the particle size exceeds these critical values (not more than 1.5 mm commercial feed), growth performance and FCR of the crayfish are no longer changed. Nevertheless, group C has high protein and low lipid content, as well as better nutrition with amino acids and fatty acids. Overall, combined with growth performance and nutrient composition, it is recommended that the particle size of the diet at the larval stage for crayfish is between 0.71 and 0.85 mm.

Application of MOF-based nanotherapeutics in light-mediated cancer diagnosis and therapy.

Light-mediated nanotherapeutics have recently emerged as promising strategies to precisely control the activation of therapeutic reagents and imaging probe both in vitro and in vivo, largely ascribed to their unique properties, including minimally invasive capabilities and high spatiotemporal resolution. Nanoscale metal-organic frameworks (NMOFs), a new family of hybrid materials consisting of metal attachment sites and bridging ligands, have been explored as a new platform for enhanced cancer diagnosis and therapy due to their tunable size, modifiable surface, good biocompatibility, high agent loading and, most significantly, their ability to be preferentially deposited in tumors through enhanced permeability and retention (EPR). Especially the light-driven NMOF-based therapeutic platform, which not only allow for increased laser penetration depth and enhanced targeting, but also enable imaging-guided or combined treatments. This review provides up-to-date developments of NMOF-based therapeutic platforms for cancer treatment with emphasis on light-triggered therapeutic strategies and introduces their advances in cancer diagnosis and therapy in recent years.

Vitamin D3 mitigates lipopolysaccharide-induced oxidative stress, tight junction damage and intestinal inflammatory response in yellow catfish, Pelteobagrus fulvidraco.

The present study explored the possible mitigative effects of vitamin D3 (VD3) on lipopolysaccharide (LPS)-induced intestinal oxidative stress, inflammatory response and tight junction damage in yellow catfish, Pelteobagrus fulvidraco. Herein, four experimental groups were established by injecting yellow catfish with NaCl, LPS, VD3 or LPS plus VD3. The results showed that LPS induced oxidative stress and that exogenous VD3 mitigated the adverse effects of LPS. Additionally, LPS suppressed the activity of antioxidant enzymes (Cat, Sod and Gr) and upregulated the mRNA expression of proinflammatory cytokines (Tnf-α, Il-1ß, Il-8). Furthermore, the mRNA expression of "fencing" tight junctions (Claudin-1, Claudin-5, Occludin, Zo-1) was downregulated, while that of "pore-forming" tight junctions (Claudin-2, Claudin-12) was upregulated, however no effect on apoptosis genes was observed (p53, Bax, Caspase-3 and Caspase-9). These LPS-induced effects were significantly reversed by pretreatment with VD3. Taken together, this study suggests that exogenous VD3 substantially alleviates LPS-induced intestinal inflammation by upregulating antioxidant activity, suppressing inflammation and promoting fencing tight junctions in the intestine.

Exogenous inositol ameliorates the effects of acute ammonia toxicity on intestinal oxidative status, immune response, apoptosis, and tight junction barriers of great blue-spotted mudskippers (Boleophthalmus pectinirostris).

Ammonia toxicity can disrupt the intestinal health of aquatic animals. It is important to find substances that alleviate these adverse effects. The present study explored the possible protective role of myo-inositol (MI) in ammonia-induced toxicity in the fish intestine. Great blue-spotted mudskippers (Boleophthalmus pectinirostris) accumulated in artificial seawater (15‰ salinity, n = 600) were randomly selected and intraperitoneally injected with NaCl (0.68%) or MI (2.5 mg/g fish in 0.68% NaCl) then exposed to artificial seawater alone (NaCl and MI group) or seawater containing 57.025 mmol/L ammonium chloride (NH3 and NH3 + MI group). After a 24-h experiment, it showed that ammonia exposure down-regulated the mRNA expression levels of intestinal barrier function proteins (Zo-1, Ocln, Cldn-5, Cldn-12, and Cldn-15) and anti-inflammatory cytokines (Tgf-ß and Il-10) while the acute ammonia stress up-regulated the apoptosis genes (p53, Bax, Caspase-3, and Caspase-9) and pro-inflammatory cytokines (Tnf-α and Il-1ß). Furthermore, ammonia challenge also induced oxidative stress, as the malondialdehyde and the protein carbonyl contents were increased. In addition, ammonia stress down-regulated the antioxidant enzymes (Cu/Zn-Sod, Cat, Gpx, and Gst) activities as well as their gene transcription levels. The administration of the exogenous myo-inositol greatly ameliorated the ammonia-induced changes in redox capacity, immune response, apoptosis, inflammation, and tight junction barrier function to levels similar to those of the NaCl group. Furthermore, fish injected with MI alone showed no significant changes compared with the NaCl group. Taken together, pretreatment with myo-inositol had no obvious side-effects and effectively protected the mudskippers' intestine from the toxicity caused by acute ammonia stress.

Effects of low salinity on gill and liver glycogen metabolism of great blue-spotted mudskippers (Boleophthalmus pectinirostris).

This study investigated the effects of low salinity exposure on glycogen and its metabolism biomarkers, glycogen synthase (GS) and glycogen phosphorylase (GP), representing glycogen synthesis and catabolism, respectively, in the gills and liver of great blue-spotted mudskippers (Boleophthalmus pectinirostris). The fish were accumulated at 10‰ salinity seawater for 1 week, then 270 healthy great blue-spotted mudskippers with similar size were randomly transferred to 10‰ (control group) or 3‰ (low salinity group) seawater for 72-hour stress experiment. Fish significantly elevated their blood glucose levels 12 h after low salinity challenge. At the end of experiments, a decrease in liver glycogen contents was observed in both the control and low salinity groups, the latter showing a pronounced decrease, while the gill glycogen contents were not changed for either group. The mRNA abundance and enzyme activity of GS and GP were both elevated in gill tissues, showing a rising glycogen synthesis and catabolism, probably resulting in the unchanging gill glycogen content. While in liver tissues, the mRNA abundance and enzyme activity were decreased for GS and increased for GP, showing a net increase for breaking down glycogen in liver, probably for supplying a sufficient glucose level for gills and other tissues/organs involved in the response to salinity changes.

Effects of α-ethinyl estradiol (EE2) and diethylhexyl phthalate (DEHP) on growth performance, antioxidant status and immune response of juvenile yellow catfish Pelteobagrus fulvidraco.

The four experimental groups were carried out to test the response of yellow catfish to EE2 and DEHP: control group was exposed to DMSO; EE2 group was exposed to 1.0 µg/L EE2; DEHP group was exposed to 1.0 mg/L DEHP; mix group was exposed to 1.0 µg/L EE2 and 1.0 mg/L DEHP. The experiment continued for 56 days. Fish survival rate was not different among experimental groups. Fish in DEHP and mix groups had the highest weight gain, and lowest value appeared in control group. The highest hepatosomatic index was found in DEHP and mix groups. Serum alanine transaminase of fish in control group was lower than other groups, but the alkaline phosphatase value was the highest. Serum total anti-oxidation capacity, superoxide dismutase and catalase activities of fish in control group were higher than other groups, but malondialdehyde content is opposite. Respiratory burst and phagocytic indices of fish in EE2 group were the lowest. After 96 h of ammonia stress, the survival rate of fish in mix group was significantly lower than control group. This study indicates that EE2 and DEHP exposure can lead to gain weight of yellow catfish, which is related to liver damage and fat accumulation; EE2 and DEHP exerts its toxic effects by inducing ROS generation, leading to lipid peroxidation and immunosuppression.

Effects of dietary glucose and starch levels on the growth, apparent digestibility, and skin-associated mucosal non-specific immune parameters in juvenile blunt snout bream (Megalobrama amblycephala).

A 10-week feeding trial was performed to evaluate the effects of different types and levels of carbohydrates in growth performance, apparent digestibility coefficients and skin-associated mucosal non-specific immune parameters in blunt snout bream (Megalobrama amblycephala). Fish were randomly fed four diets containing two carbohydrates (glucose and starch) diets and two carbohydrates levels (330 and 440 g kg-1). High carbohydrate levels remarkably increased the weight gain rate (WGR), apparent digestibility of dry matters, protein and carbohydrates, body crud protein content, plasma levels of aspartate transaminase (AST), and skin-associated mucosal levels of immunoglobulin M (IgM), HDL cholesterol, lysozyme (LZM), advanced the transcriptions of mucin 2 (Muc2), mucin 5b (Muc5b) and apolipoprotein A-I (apoA-I), whereas the opposite was true for feed conversion ratio (FCR), plasma levels of IgM, skin-associated mucosal levels of major histocompatibility complex (MHC) and ß-Defensins, and the transcriptions of heat shock protein 60 (Hsp60). In addition, carbohydrate types of glucose remarkably increased the survival rate, apparent digestibility of dry matters, protein and carbohydartes, body crud ash, plasma levels of total protein (TP), globulin (GLB), immunoglobulin M (IgM), complement C3 and complement C4 and the transcriptions of Muc5b. Whereas the carbohydrate types of starch remarkably increased viscerosomatic index (VSI), hepatosomatic index (HSI), condition factor (CF), abdominal fat percentage (AFP), apparent digestibility of liquid, advanced the transcriptions of Muc2, apoA-I and heat shock protein 70 (Hsp70). Significant interactions between different types and levels of dietary carbohydrates were also observed in WGR, apparent digestibility of dry matters, protein and liquid, body crud ash, plasma levels of TP, albumin (ALB) and AST, skin-associated mucosal levels of major histocompatibility complex (MHC) and ß-Defensins, and the transcriptions of Muc2 and Muc5b. Our results indicate that inclusion of high level of glucose in the diet of blunt snout bream could improve growth performance, nonspecific immunity, and increase the efficiency of protein, which is suggesting that high level of glucose could be used in feed production. However, the proportion of the specific formula of glucose using in feed needs further study.

Fishmeal replacement by rice protein concentrate with xylooligosaccharides supplement benefits the growth performance, antioxidant capability and immune responses against Aeromonas hydrophila in blunt snout bream (Megalobrama amblycephala).

This study aimed to investigate the effects of fishmeal (FM) replacement by rice protein concentrate (RPC) with a xylooligosaccharides supplement on the growth performance, antioxidant capability and immune response against Aeromonas hydrophila in blunt snout bream (Megalobrama amblycephala). Fish (46.85 ±â€¯0.34 g) were randomly assigned to one of 6 diets, namely the control diet (containing FM), the RPC diet (FM replaced by RPC) and RPC diet supplemented with 0.5, 1.5, 2.3 and 3% XOS respectively, for 8 weeks. After the feeding trial, fish were challenged by Aeromonas hydrophila for 96 h with the blood and liver sample obtained at 48 and 96 h respectively. The results showed that the final weight, weight gain and protein efficiency ratio of fish fed RPC diet were significantly (P < 0.05) lower than that of the control group, whereas the opposite was true for FCR. However, the supplement of 1.5% XOS remarkably (P < 0.05) improved these parameters compared to the control diet. Plasma total iron binding capacity of fish fed the RPC diet showed little difference (P > 0.05) with that of the control group, but it enhanced significantly (P < 0.05) with the supplement of 1.5% XOS compared to the control group. After bacterial infection, plasma lysozyme (LYM), complement 3, complement 4, myeloperoxidase (MPO), acid phosphatase (ACP) and alkaline phosphatase activities, as well as immunoglobulin M, levels all increased significantly (P < 0.05) with the maximum value is attained at 48 h, then they decreased significantly (P < 0.05) with further increasing time at 96 h. Similar results were also observed in liver superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activities as well as malondialdehyde (MDA) content. Regarding dietary treatment, these parameters of fish fed RPC diet showed little difference (P > 0.05) with those fed the control diet but were significantly enhanced (P < 0.05) when RPC diet was supplemented with 1.5-2.3% XOS, except for hepatic (MDA) content which showed an opposite trend compared to the control group. After 96 h of challenge, the relative percentage survival (RPS) of fish fed XOS was significantly higher (P < 0.05) than that of fish offered the control and RPC diet. In addition, significant (P < 0.05) interactions between dietary XOS and sampling time were also observed in plasma LYM, MPO, ACP, and hepatic SOD, CAT, GPX, MDA, as well as RPS. In conclusion, this study indicated that dietary supplementation of 1.5%XOS could significantly improve the growth performance, antioxidant capability, innate immunity and A. hydrophila resistance of blunt snout bream fed diets with FM replaced by RPC.

Molecular characterization of AMP-activated protein kinase α2 from herbivorous fish Megalobrama amblycephala and responsiveness to glucose loading and dietary carbohydrate levels.

This study aimed to characterize the full-length cDNA of AMPKα2 in Megalobrama amblycephala, and evaluate its potential role in glucose homeostasis and carbohydrate metabolism. The cDNA obtained covered 1942bp with an open reading frame of 1635bp encoding 545 amino acids. Multiple alignments and phylogenetic analysis revealed a high homology (91-100%) among most fish and higher vertebrates. This AMPKα2 mRNA predominantly expressed in muscle, liver and brain, while little in gill and intestine. Then, the AMPKα2 expressions were determined in the muscle, liver and brain of fish subjected to a glucose load (injected intraperitoneally with 0, 1.67 and 3.34g glucose per kg body weight) and after a 12-week feeding trial (fed two dietary carbohydrate levels: 30% and 43%), respectively. After the glucose load, plasma glycemia peaked at 1h in fish. Thereafter, it decreased significantly to the basal level at 8h. However, AMPKα2 expression in muscle, liver and brain all decreased significantly during the first 2h, then returned to the basal value at 24h. Unlikely, tissue AMPKα2 expression of fish receiving saline solution increased significantly during the whole sampling period. Additionally, high-carbohydrate diet enhanced its expression in liver and muscle, but not that in brain. These findings indicated that the AMPKα2 gene shared a high degree of conservation with that of the other vertebrates. Muscle, liver and brain AMPKα2 expressions were highly induced by glucose administration. Furthermore, high dietary carbohydrate modified its expressions in these tissues.

Effects of emodin and vitamin E on the growth and crowding stress of Wuchang bream (Megalobrama amblycephala).

This study aimed to investigate the effects of dietary emodin, high-dose vitamin E and their combination on the growth of Megalobrama amblycephala and its resistance to acute crowding stress. The fish were randomly divided into four groups: a control group fed with basal diet, and three treatment groups fed with basal diet supplemented with 60 mg/kg emodin (the emodin group), 500 mg/kg vitamin E (the vit E group), and 60 mg/kg emodin together with 500 mg/kg vitamin E (the combination group). After 60 days, the fish were exposed to acute crowding stress for 24 h. The results showed that the weight gain of the vit E group, specific growth rate of the vit E group, total serum protein concentration (TP) of the vit E group, serum lysozyme activity of the emodin group, serum superoxide dismutase (SOD) activity of the emodin group, hepatic heat shock protein 70 (HSP70) levels of the vit E group and the emodin group, and serum alanine aminotransferase (ALT) activity of the combination group significantly increased while the weight gain and specific growth rate of the combination group significantly decreased compared with the control group before stress. After crowding stress, the vit E group had improved serum TP 12 h post-stress, hepatic SOD activity 24 h post-stress, and hepatic HSP70 mRNA levels 12 and 24 h post-stress while the emodin group had enhanced serum SOD activity 12 and 24 h post-stress and hepatic HSP70 mRNA levels 12 and 24 h post-stress, as compared with the control. However, the serum cortisol content of the three treatment groups 12 and 24 h post-stress, ALT activity in the vit E group and emodin group 24 h post-stress, and serum alkaline phosphatase and liver catalase activity in the combination group 24 h post-stress were lower than those in the control group. The cumulative mortality was lower in the emodin, vit E, and combination group after Aeromonas hydrophila infection compared with the control group. Therefore, dietary supplementation with 60 mg/kg emodin or 500 mg/kg vitamin E can improve HSP70 mRNA levels and antioxidant capabilities, resistance to crowding stress, and growth in M. amblycephala. However, the combination of emodin and vit E does not have a synergistic effect in M. amblycephala.

The influence of various feeding patterns of emodin on growth, non-specific immune responses, and disease resistance to Aeromonas hydrophila in juvenile Wuchang bream (Megalobrama amblycephala).

The present study was conducted to evaluate the effect of various feeding patterns of emodin on growth, non-specific immune response, and disease resistance to Aeromonas hydrophila in juvenile Wuchang bream. Healthy Megalobrama amblycephala (initial weight: 3.47 ± 0.032) were grown in a circulating water system for 8 weeks. Five groups were studied: one control group was fed with a basal diet for eight weeks (Pattern 1, P1), and three treatment groups were fed with a trial diet of 30 mg emodin kg(-1) at one-week (Pattern 2, P2), two-week (Pattern 3, P3), four-week (Pattern 4, P4) intervals. The final treatment group maintained the trial diet for the entire eight-week study duration (Pattern 5, P5). Results indicated that different feeding patterns of emodin significantly influenced the weight gain rate of Wuchang bream (P < 0.05). Fish in the P4 treatment group had significantly higher rates of weight gain (WG) than those in other treatment groups. There were no significant differences in survival rates or feed conversion ratios (FCR) between treatment groups and the control group. White blood cell count (WBC), respiratory burst activity, superoxide dismutase (SOD) activity, myeloperoxidase (MPO) activity and tumor necrosis factor-α (TNF-α) activity were shown to increase at first and then decrease from P3 condition to P5 condition. Fish under P4 treatment showed the most significant improvement of all tested parameters compared to control. Significantly higher levels (P < 0.05) of plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity were observed in P2 and P4 treatment groups when compared with the control group, while no significant differences were observed in the AST and ALT activity of fish in P2, P3, P4 and P5 treatment groups. In a bacterial challenge experiment with A. hydrophila, fish under P4 and P5 treatment showed lower cumulative mortality than the control group. The results of this study suggest that an initial 4-week feeding interval is recommended for the economic and practical culture of M. amblycephala.