Effect of different concentrations of gibberellins on attenuation of nutrient and antibiotics from aquaculture wastewater using microalgae-bacteria-fungi consortia system.

This study assessed the effect of gibberellins (GAs) concentrations on antibiotic and nutrient removal using diverse microalgal-bacterial-fungal consortia. Five systems (Chlorella vulgaris, T1; C. vulgaris + S395-2 + Clonostachys rosea, T2; C. vulgaris + S395-2 + Ganoderma lucidum, T3; C. vulgaris + S395-2 + Pleurotus pulmonarius, T4; and C. vulgaris + S395-2, T5) were established, and optimal conditions and effective symbiosis were applied to improve antibiotic and nutrient removal. Consortium growth was T2 > T3 > T5 > T4 > T1, while GA impact ranked 50 mg L-1 > 20 mg L-1 > 80 mg L-1 > 0 mg L-1. After 7 days at 50 mg L-1 GAs, total nitrogen (TN), NH4-N, NO3-N, and total phosphorous (TP) removal reached 85.97 %, 78.08 %, 86.59 %, and 94.39 %, respectively. Florfenicol, oxytetracycline hydrochloride, ofloxacin, and sulfamethoxazole removal efficiencies were 67.77 %, 98.29 %, 90.47 %, and 94.92 %, respectively. These findings highlight GAs' significant role in enhancing antibiotic and nutrient removal.

A Novel Disease (Water Bubble Disease) of the Giant Freshwater Prawn Macrobrachium rosenbergii Caused by Citrobacter freundii: Antibiotic Treatment and Effects on the Antioxidant Enzyme Activity and Immune Responses.

The giant freshwater prawn, Macrobrachium rosenbergii, is an important and economical aquaculture species widely farmed in tropical and subtropical areas of the world. A new disease, "water bubble disease (WBD)", has emerged and resulted in a large loss of M. rosenbergii cultured in China. A water bubble with a diameter of about 7 mm under the carapace represents the main clinical sign of diseased prawns. In the present study, Citrobacter freundii was isolated and identified from the water bubble. The optimum temperature, pH, and salinity of the C. freundii were 32 °C, 6, and 1%, respectively. A challenging experiment showed that C. freundii caused the same typical signs of WBD in prawns. Median lethal dose of the C. freundii to prawn was 104.94 CFU/g. According to the antibiogram tests of C. freundii, florfenicol and ofloxacin were selected to evaluate their therapeutic effects against C. freundii in prawn. After the challenge with C. freundii, 86.67% and 72.22% survival of protective effects against C. freundii were evaluated in the oral florfenicol pellets and oral ofloxacin pellets feding prawns, respectively, whereas the mortality of prawns without fed antibiotics was 93%. After antibiotic treatment and C. freundii infection, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), malondialdehyde (MDA), acid phosphatase (ACP), alkaline phosphatase (ALP), and lysozyme (LZM) in the hemolymph and hepatopancreas of the prawns and the immune-related gene expression levels of Cu/Zn-SOD, CAT, GPx, GST, LZM, ACP, anti-lipopolysaccharide factor, crustin, cyclophilin A, and C-type lectin in hepatopancreas were all significantly changed, indicating that innate immune responses were induced by C. freundii. These results can be beneficial for the prevention and control of C. freundii in prawns.

High-Carbohydrate Diet Alleviates the Oxidative Stress, Inflammation and Apoptosis of Megalobrama amblycephala Following Dietary Exposure to Silver Nanoparticles.

A 12-week feeding trial was performed to evaluate the effects of high-carbohydrate diet on oxidative stress, inflammation and apoptosis induced by silver nanoparticles (Ag-NPs) in M. amblycephala. Fish (20.12 ± 0.85 g) were randomly fed four diets (one control diet (C, 30% carbohydrate), one control diet supplemented with 100 mg kg-1 Ag-NPs (CS), one high-carbohydrate diet (HC, 45% carbohydrate) and one HC diet supplemented with 100 mg kg-1 Ag-NPs (HCS)). The results indicated that weight gain rate (WGR), specific growth rate (SGR), antioxidant enzyme (SOD and CAT) activities and expression of Trx, Cu/Zn-SOD, Mn-SOD, CAT and GPx1 of fish fed CS diet were all remarkably lower than those of other groups, whereas the opposite was true for plasma IL 1ß and IL 6 levels, liver ROS contents, hepatocytes apoptotic rate, AMP/ATP ratio, AMPKα, P 53 and caspase 3 protein contents and mRNA levels of AMPKα 1, AMPKα 2, TXNIP, NF-κB, TNFα, IL 1ß, IL 6, P 53, Bax and caspase 3. However, high-carbohydrate diet remarkably increased WGR, SGR, liver SOD and CAT activities, AMPKα protein content and mRNA levels of antioxidant genes (Cu/Zn-SOD, Mn-SOD, CAT and GPx1), anti-inflammatory cytokines (IL 10) and anti-apoptotic genes (Bcl 2) of fish facing Ag-NPs compared with the CS group, while the opposite was true for liver ROS contents, hepatocytes apoptotic rate, P 53 and caspase 3 protein contents, as well as mRNA levels of TXNIP, NF-κB, TNFα, IL 1ß, IL 6, P 53, Bax and caspase 3. Overall, high-carbohydrate diet could attenuate Ag-NPs-induced hepatic oxidative stress, inflammation and apoptosis of M. amblycephala through AMPK activation.

A transcriptome study on Macrobrachium nipponense hepatopancreas experimentally challenged with white spot syndrome virus (WSSV).

White spot syndrome virus (WSSV) is one of the most devastating pathogens of cultured shrimp, responsible for massive loss of its commercial products worldwide. The oriental river prawn Macrobrachium nipponense is an economically important species that is widely farmed in China and adult prawns can be infected by WSSV. However, the molecular mechanisms of the host pathogen interaction remain unknown. There is an urgent need to learn the host pathogen interaction between M. nipponense and WSSV which will be able to offer a solution in controlling the spread of WSSV. Next Generation Sequencing (NGS) was used in this study to determin the transcriptome differences by the comparison of control and WSSV-challenged moribund samples, control and WSSV-challenged survived samples of hepatopancreas in M. nipponense. A total of 64,049 predicted unigenes were obtained and classified into 63 functional groups. Approximately, 4,311 differential expression genes were identified with 3,308 genes were up-regulated when comparing the survived samples with the control. In the comparison of moribund samples with control, 1,960 differential expression genes were identified with 764 genes were up-regulated. In the contrast of two comparison libraries, 300 mutual DEGs with 95 up-regulated genes and 205 down-regulated genes. All the DEGs were performed GO and KEGG analysis, overall a total of 85 immune-related genes were obtained and these gene were groups into 13 functions and 4 KEGG pathways, such as protease inhibitors, heat shock proteins, oxidative stress, pathogen recognition immune receptors, PI3K/AKT/mTOR pathway, MAPK signaling pathway and Ubiquitin Proteasome Pathway. Ten genes that valuable in immune responses against WSSV were selected from those DEGs to furture discuss the response of host to WSSV. Results from this study contribute to a better understanding of the immune response of M. nipponense to WSSV, provide information for identifying novel genes in the absence of genome of M. nipponense. Furthermore, large number of transcripts obtained from this study could provide a strong basis for future genomic research on M. nipponense.

Experimental inoculation of oriental river prawn Macrobrachium nipponense with white spot syndrome virus (WSSV).

The oriental river prawn Macrobrachium nipponense is an economically important species that is widely farmed in China. White spot syndrome virus (WSSV) is one of the most devastating pathogens of the cultured shrimp Litopenaeus vannamei, responsible for massive loss of its commercial products worldwide. We investigated the infectivity and pathogenicity of WSSV in adult M. nipponense using standardized conditions for L. vannamei. The median lethal dose of WSSV in adult M. nipponense was 103.84±0.06 copies g-1, which was about 1000-fold higher than in L. vannamei (100.59±0.22 copies g-1). WSSV was detected by 2-step PCR in the gills, hepatopancreas, muscle, stomach, heart, gut, nerve, integument, pereopod, eyestalk, testis, and ovary of experimentally infected dead M. nipponense. Lesions were observed histologically following WSSV injection, showing basophilic intranuclear inclusion bodies in the hepatopancreas and subsequently in the gills. The clearance of WSSV was observed in hepatopancreas and gills at 48 and 96 h post-inoculation, respectively. No histological lesions were detected in muscle from 0-96 h post-injection. The results show that the oriental river prawn M. nipponense can be infected by WSSV and the infections are self limiting over time; therefore, M. nipponense may serve as a useful model for studying resistance to WSSV.

Anti-inflammatory and hepatoprotective effects of Ganoderma lucidum polysaccharides on carbon tetrachloride-induced hepatocyte damage in common carp (Cyprinus carpio L.).

The aim of this study was to investigate the anti-inflammatory and hepatoprotective effects of Ganoderma lucidum polysaccharides (GLPS) on carbon tetrachloride (CCl4)-induced hepatocyte damage in common carp (Cyprinus carpio L.). GLPS (0.1, 0.3, 0.6mg/ml) were added to the primary hepatocytes before (pre-treatment), after (post-treatment) and both before and after (pre- and post-treatment) the incubation of the hepatocytes with CCl4 at the concentration of 8mM in the culture medium. The supernatants and cells were collected respectively to detect the biochemical indicators. The levels of TNF-α, IL-1ß, caspase-3 and caspase-8 were measured by ELISA, the mRNA expressions of CYP1A and CYP3A were determined by RT-PCR, and western blotting was used to assay the relative protein expressions of c-Rel and p65. Results showed that GLPS significantly improved cell viability and inhibited the elevations of the marker enzymes (GOT, GPT, LDH) and MDA induced by CCl4, and markedly increased the level of SOD. Treatments with GLPS resulted in a significant decrease in the expressions of CYP1A and CYP3A, and significantly down-regulated extrinsic apoptosis and immune inflammatory response. In brief, the present study showed that GLPS can protect hepatocyte injury induced by CCl4 through inhibiting lipid peroxidation, elevating antioxidant enzyme activity and suppressing apoptosis and immune inflammatory response.

Effects of curcumin on antioxidative activities and cytokine production in Jian carp (Cyprinus carpio var. Jian) with CCl4-induced liver damage.

We investigated the protective effects of curcumin on liver-damaged Cyprinus carpio var. Jian (Jian carp). The carp were fed 0.1%, 0.5%, or 1.0% curcumin for 60 days, then injected intraperitoneally with 30% carbon tetrachloride solution. Liver and blood samples were collected to measure the liver index, serum- and liver-associated enzymes, liver histology, nuclear factor-κB (NF-κB)/c-Rel, interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), and IL-12 mRNA expression, and the level of NF-κB/c-Rel protein in the liver, and for a comet assay. We found that 0.5% and 1.0% curcumin significantly reduced the CCl(4)-induced increase in the liver index. The comet assay showed that the tail moment, olive tail moment, tail length, and tail DNA% improved in fish pretreated with 0.5 or 1.0% curcumin. CCl(4)-induced histological changes, including extensive hepatocyte degeneration, indistinct cell borders, nuclear condensation, and karyolysis were clearly reduced after treatment with 0.5% and 1.0% curcumin. Moreover, 0.5% and 1.0% curcumin significantly inhibited the CCl(4)-induced increase in serum glutamic oxaloacetic transaminase and promoted the restoration of superoxide dismutase in the liver; 1.0% curcumin significantly reduced serum glutamic pyruvic transaminase and lactate dehydrogenase and hepatic malondialdehyde, but significantly increased the total antioxidant capacity and glutathione levels in the liver. The CCl(4)-induced upregulation of NF-κB/c-Rel, IL-1ß, and TNF-α mRNAs and NF-κB/c-Rel protein levels was inhibited by 0.5% and 1.0% curcumin, and IL-12 mRNA was reduced by all three doses of curcumin. The effects of curcumin on the liver index, enzymes, histological changes, and cytokines were dose-dependent. Our results indicate that curcumin reduces CCl(4)-induced liver damage in Jian carp by upregulating antioxidative activities and inhibiting NF-κB, IL-1ß, TNF-α, and IL-12 expression.